SULFONANILIDE AND BENZYLSULFONYL DERIVATIVES, AND COMPOSITIONS

AND METHODS THEREOF

Priority Claims and Related Patent Applications

[0001] This application claims the benefit of priority from U.S. Provisional Application Serial Nos. 63/088,158, filed on October 6, 2020, and 63/110,153, filed on November 5, 2020, the entire content of each of which is incorporated herein by reference in its entirety.

Technical Field of the Invention

[0002] The invention generally relates to novel compounds and therapeutic uses thereof. More particularly, the invention provides novel sulfonanilide and benzylsulfonyl derivatives, their salts, solvates, hydrates and polymorphs thereof as transient receptor potential cation channel, mucolipin subfamily (TRPML) modulators. The invention also provides pharmaceutical compositions comprising a compound of the invention and methods thereof for treating various diseases and disorders associated with or related to TRPML activities such as lysosome storage diseases, muscular dystrophy, age-related common neurodegenerative diseases, reactive oxygen species (ROS) or oxidative stress related diseases, and ageing.

Background of the Invention

[0003] The lysosome, the cell’s recycling center, can mediate the degradation of a variety of biomaterials (proteins, lipids, and membranes) into smaller molecules or building blocks, which will be subsequently transported out of lysosomes for reutilization or energy (see, e.g., de Duve 2005 Nat Cell Biol 7 (9): 847-9; Parkinson-Lawrence, et al. 2010 Physiology (Bethesda) 25(2): 102-15). Problems in either the degradation step (due to lack of hydrolytic enzymes) or the transport step lead to lysosome storage (of accumulated materials) and more than 50 human diseases collectively called lysosome storage diseases (LSDs). Lysosome storage can in turn affect lysosomal degradation and membrane transport/trafficking, making a positive feedback loop and a vicious cycle. Because lysosome storage is also seen in common neurodegenerative diseases such as Alzheimer’s and Parkinson’s, understanding the mechanisms underlying the positive feedback loop may provide therapeutic approaches not only for LSDs, but also for common sporadic neurodegenerative diseases. A lysosome-localized Ca ²⁺ channel, TRPML1, has been recently identified as a key regulator of most membrane trafficking processes in the lysosome. Human mutations of TRPML1 cause lysosomal trafficking defects, lysosome storage, and neurodegenerative diseases.

[0004] TRPML1 (also abbreviated as ML1), a member of the TRP-type Ca ²⁺ channel superfamily, is the principle Ca ²⁺ channel in the lysosome (see, e.g., Cheng, et al. 2010 FEBS Lett 584(10): 2013- 21). Loss-of-function mutations in the human TRPML1 gene cause Type IV Mucolipidosis (ML4), a lysosome storage neurodegenerative disease. TRPML1 ^{- -} (abbreviated as MLl ^_/ “) skin fibroblasts from ML4 patients are characterized by the accumulation of enlarged endosomal/lysosomal compartments (vacuoles) in which lipids and other biomaterials build up, suggestive of trafficking defects. Analyses of trafficking kinetics suggest that the primary defects are in the late endocytic pathways. First, ML1 is likely to be required for the formation of transport vesicles from the LEL to the Trans-Golgi Network (TGN) (LEL-to-TGN retrograde trafficking). Second, fusion of lysosomes with the plasma membrane (referred to as lysosomal exocytosis), a process that is important in cellular waste elimination, membrane repair, and phagocytosis, is defective in ML4 cells. Defects in either trafficking steps could lead to lysosome storage. Because the release of Ca ²⁺ from lysosomes (lysosomal Ca ²⁺ release) is essential for both trafficking steps, it is hypothesized that ML1 is indeed the Ca ²⁺ release channel that regulates lysosomal trafficking.

[0005] PI(3,5)P2 is a low-abundance phosphoinositide, is the primary activator of ML1, and a positive regulator of lysosomal trafficking. Both TRPML1 -lacking and PI(3,5)P2-deficient cells exhibit defects in LEL-to-Golgi retrograde trafficking and autophagosome-lysosome fusion, suggesting that the TRPML1-PI(3,5)P2 system represents a common signaling pathway essential for late endocytic trafficking.

[0006] Due to the function of lysosome in lysosomal trafficking, lysosomes are required for quality-control regulation of mitochondria, the “power house” of the cell and the major source of endogenous ROS (reactive oxygen species). Damaged mitochondria causes oxidative stress, which is a common feature of most LSDs, common neurodegenerative diseases, and ageing (Xu, et al. 2015 Annu Rev Physiol 77, 57-80). Recent studies suggest that mitochondria are localized in close physical proximity to lysosomes (Elbaz-Alon, et al. 2014 Dev Cell 30, 95-102; Li, et al. 2015 Cell Mol Neurobiol 35, 615-621). Hence the lysosomal membrane is potentially an accessible and direct target of ROS signaling. Given that ROS reportedly regulate ion channels (Bogeski, et al. 2014 Antioxid Redox Signal 21, 859-862), it is possible that lysosomal conductances, particularly through lysosomal Ca ²⁺ channels such as TRPML1, may mediate ROS-regulation of lysosomal function. Indeed, electrophysiological studies revealed that whole-endolysosome TRPML1 currents were directly activated by ROS.

[0007] A regulatory imbalance can result in elevated ROS levels and oxidative stress, which are believed to underlie a variety of metabolic and neuro degenerative diseases, as well as ageing (Barnham c/ a/. 2004 Nat Rev DrugDiscov 3, 205-214; Scherz-Shouval, et al. 2011 Trends Biochem Sci 36, 30-38). Given the role of TRPML1 in mediating ROS-induced autophagy, a TRPML1 agonist might be able to clear the excessive ROS, thereby ameliorating the ROS related diseases and ageing, especially photo ageing in the skin.

[0008] Transcription factor (TF)EB regulates autophagy and lysosome biogenesis. Overexpression of TFEB has been reportedly induce cellular clearance in a number of lysosome storage diseases, including Pombe Disease, Cystinosis, multiple sulfatase deficiency, as well as common neurodegenerative diseases, including Parkinson’s disease and Huntinton’s disease (Settembre, et al., 2013 Nat Rev Mol Cell Biol 14(5), 283-96). Therefore, activation of TRPML1 by TRPML1 agonists may also lead to cellular clearance in all the aforementioned diseases, providing therapeutic targets for these devastating diseases.

[0009] Recently, a potent synthetic agonist for TRPML1 has been reported (Shen, et al. 2012 Nat Commun 3, 731). This SF-51-related compound (Mucolipin Synthetic Agonist 1 or ML-SA1) that could induce significant [Ca2+] _cyt increases in HEK293 cells stably or transiently expressing ML1- 4A. In electrophysiological assays, ML-SA1 robustly activated whole-cell ZMLWA and whole- endolysosome ZMLL ML-SA1 also activated whole-cell ZTRPML2 and ZTRPMLS, but not six other related channels. ML-SA1 (10 μM) activation of whole-endolysosome ZMLI was comparable to the effect of the endogenous TRPML agonist PI(3,5)P2 (IμM), and these agonists were synergistic with each other. ML-SA1 activated an endogenous whole-endolysosome TRPML-like current (ZML-L) in all mammalian cell types that were investigated, including Chinese Hamster Ovary (CHO), Cos-1, HEK293, skeletal muscle, pancreatic P and macrophage cells. ML-SA1 activated whole- endolysosome ZML L in wild-type (WT; ML1 ^+/+ ), but not ML4 (MLI ^z ) human fibroblasts, suggesting that although ML-SA1 targets all three TRPMLs, the expression levels of TRPML2 and TRPML3 are very low, and TRPML 1 is the predominant lysosomal TRPML channel in this cell type. These results suggest that ML-SA1 is a reasonably specific and potent agonist that can be a useful for modulating the functions of TRPMLs.

[0010] High concentrations of ML-SA1 (~10 μM) are needed to effectively activate TRPMLs. Since that concentration is usually difficult to achieve in vivo, ML-SA1 cannot be used to treat the above TRPML related diseases.

[0011] There is an urgent need for more potent TRPML activators, in particular, compounds that are useful in treating disorders related to TRPML activities such as lysosome storage diseases, muscular dystrophy, age-related common neurodegenerative diseases, ROS or oxidative stress related diseases, and ageing.

Summary of the Invention

[0012] The invention is based in part on novel sulfonanilide and benzylsulfonyl-containing compounds, pharmaceutical compositions thereof and methods of their preparation and use in treating or reducing various diseases or disorders. In particular, compounds, compositions and methods of the invention are useful in treating diseases or disorders mediated by or associated with TRPMLs.

[0013] In one aspect, the invention generally relates to a compound having the structural formula I: or a pharmaceutically acceptable form or an isotope derivative thereof, wherein:

R ¹ is a 5-membered heteroaryl substituted with 1, 2, or 3 substituents at least one of which is selected from the group consisting of C _1-5 alkyl, CF ₃ , CH ₂ CF ₃ , CN, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR’; and wherein any alkyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, halo, OH, OC _1-3 alkyl, and CN;

R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 4- to 8-membered heterocyclyl which optionally comprises a further heteroatom selected from the group consisting of nitrogen optionally substituted with alkyl or aryl, oxygen and S(O) _n , wherein said 4- to 8-membered heterocyclyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8- membered heterocyclyl spiro fused to a C _3.7 cycloalkyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8-membered bridged heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; each of R ⁴ and R ⁵ is independently H, C _1-3 alkyl, halogen, oxo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, or S(O) ₂ NRR’; each of R and R’ is independently H, or C _1-5 alkyl or, optionally, R and R’, together with the nitrogen to which they are attached to, form a 4- to 6- member ring;

X is CR ⁶ R ⁷ , or NR ⁶ , wherein each of R ⁶ and R ⁷ is independently H, or C1-C3 alkyl; and n is 0, 1 or 2.

[0014] In another aspect, the invention generally relates to a pharmaceutical composition comprising a compound disclosed herein, effective to treat or reduce one or more diseases or disorders, in a mammal, including a human, and a pharmaceutically acceptable excipient, carrier, or diluent. [0015] In yet another aspect, the invention generally relates to a pharmaceutical composition comprising an amount of a compound of formula I: or a pharmaceutically acceptable form or an isotope derivative thereof, wherein:

R ¹ is a 5-membered heteroaryl substituted with 1, 2, or 3 substituents at least one of which is selected from the group consisting of C _1-5 alkyl, CF ₃ , CH ₂ CF ₃ , CN, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR’; and wherein any alkyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, halo, OH, OC _1-3 alkyl, and CN;

R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 4- to 8-membered heterocyclyl which optionally comprises a further heteroatom selected from the group consisting of nitrogen optionally substituted with alkyl or aryl, oxygen and S(O) _n , wherein said 4- to 8-membered heterocyclyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8- membered heterocyclyl spiro fused to a C3.7 cycloalkyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8-membered bridged heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; each of R ⁴ and R ⁵ is independently H, C _1-3 alkyl, halogen, oxo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, or S(O) ₂ NRR’; each of R and R’ is independently H, or C _1-5 alkyl or, optionally, R and R’, together with the nitrogen to which they are attached to, form a 4- to 6- member ring;

X is CR ⁶ R ⁷ , or NR ⁶ , wherein each of R ⁶ and R ⁷ is independently H, or C ₁ -C ₃ alkyl; and n is 0, 1 or 2, effective to treat, or reduce one or more diseases or disorders, in a mammal, including a human, and a pharmaceutically acceptable excipient, carrier, or diluent.

[0016] In yet another aspect, the invention generally relates to a unit dosage form comprising a pharmaceutical composition disclosed herein.

[0017] In yet another aspect, the invention generally relates to a method for treating or reducing a disease or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

[0018] In yet another aspect, the invention generally relates to a method for treating or reducing the effect of aging comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

[0019] In yet another aspect, the invention generally relates to a method for treating or reducing oxidative stress or reactive oxygen species related diseases or disorder, comprising administering to a subject in need thereof an effective amount of a TRPML1 agonist or a composition comprising of a TRPML1 agonist.

[0020] In yet another aspect, the invention generally relates to a method for treating or reducing oxidative stress or reactive oxygen species related diseases or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

[0021] In yet another aspect, the invention generally relates to use of a compound disclosed herein, and a pharmaceutically acceptable excipient, carrier, or diluent, in preparation of a medicament for treating a disease or disorder.

Definitions

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. General principles of organic chemistry, as well as specific functional moieties and reactivity, are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 2006.

[0023] The following terms, unless indicated otherwise according to the context wherein the terms are found, are intended to have the following meanings. [0024] Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 16 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

[0025] Any compositions or methods disclosed herein can be combined with one or more of any of the other compositions and methods provided herein.

[0026] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0027] Definitions of specific functional groups and chemical terms are described in more detail below. When a range of values is listed, it is intended to encompass each value and sub-range within the range. For example, “C _1-6 alkyl” is intended to encompass, C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C _1-6 , C _1-5 , C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-5 , C _2-4 , C _2-3 , C _3-6 , C _3-5 , C _3-4 , C _4-6 , C _4-5 , and C _5-6 alkyl.

[0028] Where substituent groups are specified by their conventional chemical formulae, written from left to right, they equally encompass the chemically identical substituents that would result from writing the structure from right to left, e.g., -C(=O)-O- is equivalent to -O-C(=O)-.

[0029] Structures of compounds of the invention are limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to give compounds that are not inherently unstable and/or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions (e.g., aqueous, neutral, and several known physiological conditions).

[0030] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural reference, unless the context clearly dictates otherwise.

[0031] As used herein, “at least” a specific value is understood to be that value and all values greater than that value.

[0032] As used herein, the terms “comprises,” “comprising”, or "having" when used to define compositions and methods, are intended to mean that the compositions and methods include the recited elements, but do not exclude other elements. The term “consisting essentially of’, when used to define compositions and methods, shall mean that the compositions and methods include the recited elements and exclude other elements of any essential significance to the compositions and methods. For example, “consisting essentially of’ refers to administration of the pharmacologically active agents expressly recited and excludes pharmacologically active agents not expressly recited. The term consisting essentially of does not exclude pharmacologically inactive or inert agents, e.g., pharmaceutically acceptable excipients, carriers or diluents. The term “consisting of’, when used to define compositions and methods, shall mean excluding trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

[0033] As used herein, the terms “disease” and “disorder” are used interchangeably and refer to any condition that damages or interferes with the normal function of a cell, tissue, or organ.

[0034] As used herein, the term “hydrate” means a compound which further includes a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces. [0035] As used herein, the term "pharmaceutically acceptable” refers to being suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. A "pharmaceutically acceptable form" of a disclosed compound includes, but is not limited to, pharmaceutically acceptable salts, esters, hydrates, solvates, polymorphs, isomers, prodrugs, and isotopically labeled derivatives thereof. In one embodiment, a "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable salts, esters, prodrugs and isotopically labeled derivatives thereof. In some embodiments, a "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable isomers and stereoisomers, prodrugs and isotopically labeled derivatives thereof.

[0036] In certain embodiments, the pharmaceutically acceptable form is a pharmaceutically acceptable salt. As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of subjects without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds provided herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, besylate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2- hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3 -phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. In some embodiments, organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoracetic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0037] The salts can be prepared in situ during the isolation and purification of the disclosed compounds, or separately, such as by reacting the free base or free acid of a parent compound with a suitable base or acid, respectively. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (Ci-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, such as isopropyl amine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt can be chosen from ammonium, potassium, sodium, calcium, and magnesium salts.

[0038] In certain embodiments, the pharmaceutically acceptable form is a "solvate" (e.g., a hydrate). As used herein, the term "solvate" refers to compounds that further include a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermol ecul ar forces. The solvate can be of a disclosed compound or a pharmaceutically acceptable salt thereof. Where the solvent is water, the solvate is a "hydrate". Pharmaceutically acceptable solvates and hydrates are complexes that, for example, can include 1 to about 100, or 1 to about 10, or 1 to about 2, about 3 or about 4, solvent or water molecules. It will be understood that the term "compound" as used herein encompasses the compound and solvates of the compound, as well as mixtures thereof.

[0039] In certain embodiments, the pharmaceutically acceptable form is a prodrug. As used herein, the term "prodrug" (or “pro-drug”) refers to compounds that are transformed in vivo to yield a disclosed compound or a pharmaceutically acceptable form of the compound. A prodrug can be inactive when administered to a subject, but is converted in vivo to an active compound, for example, by hydrolysis (e.g., hydrolysis in blood). In certain cases, a prodrug has improved physical and/or delivery properties over the parent compound. Prodrugs can increase the bioavailability of the compound when administered to a subject (e.g., by permitting enhanced absorption into the blood following oral administration) or which enhance delivery to a biological compartment of interest (e.g., the brain or lymphatic system) relative to the parent compound. Exemplary prodrugs include derivatives of a disclosed compound with enhanced aqueous solubility or active transport through the gut membrane, relative to the parent compound.

[0040] The prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in a mammalian organism (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7- 9, 21-24 (Elsevier, Amsterdam). A discussion of prodrugs is provided in Higuchi, T., et al., "Prodrugs as Novel Delivery Systems," A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated in full by reference herein. Exemplary advantages of a prodrug can include, but are not limited to, its physical properties, such as enhanced water solubility for parenteral administration at physiological pH compared to the parent compound, or it can enhance absorption from the digestive tract, or it can enhance drug stability for long-term storage.

[0041] Prodrugs commonly known in the art include well-known acid derivatives, such as, for example, esters prepared by reaction of the parent acids with a suitable alcohol, amides prepared by reaction of the parent acid compound with an amine, basic groups reacted to form an acylated base derivative, etc. Of course, other prodrug derivatives may be combined with other features disclosed herein to enhance bioavailability. As such, those of skill in the art will appreciate that certain of the presently disclosed compounds having free amino, amido, hydroxy or carboxylic groups can be converted into prodrugs. Prodrugs include compounds having an amino acid residue, or a polypeptide chain of two or more (e.g., two, three or four) amino acid residues which are covalently joined through peptide bonds to free amino, hydroxy or carboxylic acid groups of the presently disclosed compounds. The amino acid residues include the 20 naturally occurring amino acids commonly designated by three letter symbols and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3 -methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid, citrulline homocysteine, homoserine, ornithine and methionine sulfone. Prodrugs also include compounds having a carbonate, carbamate, amide or alkyl ester moiety covalently bonded to any of the above substituents disclosed herein.

[0042] Particularly favored prodrugs and prodrug salts are those that increase the bio avail ability of the compounds of this invention when such compounds are administered to a mammal (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment (e.g., the brain or central nervous system) relative to the parent species. Examples of prodrugs include derivatives where a group that enhances aqueous solubility or active transport through the gut membrane is appended to the structure of formulae described herein. (See, e.g., Alexander, et al. 1988 J Med Chem 31, 318- 322; Bundgaard, et al. 1985 Elsevier: Amsterdam 1-92; Bundgaard, et al. 1987 J Med Chem 30, 451- 454; Bundgaard, H. A Textbook of Drug Design and Development; Harwood Academic Publ.: Switzerland, 1991, 113-191; Digenis, et al. Handbook of Experimental Pharmacology 1975, 28, 86- 112; Friis, et al. Textbook of Drug Design and Development; 2 ed.; Overseas Publ.: Amsterdam, 1996, 351-385; Pitman 1981 Medicinal Research Reviews 1, 189-214.)

[0043] As used herein, the term “pharmaceutically acceptable” excipient, carrier, or diluent refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate, magnesium stearate, and polyethylene oxide-polypropylene oxide copolymer as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0044] As used herein, the term “polymorph” means solid crystalline forms of a compound or complex thereof which may be characterized by physical means such as, for instance, X-ray powder diffraction patterns or infrared spectroscopy. Different polymorphs of the same compound can exhibit different physical, chemical and/or spectroscopic properties. Different physical properties include, but are not limited to stability (e.g., to heat, light or moisture), compressibility and density (important in formulation and product manufacturing), hygroscopicity, solubility, and dissolution rates (which can affect bioavailability). Differences in stability can result from changes in chemical reactivity (e.g., differential oxidation, such that a dosage form discolors more rapidly when comprised of one polymorph than when comprised of another polymorph) or mechanical characteristics (e.g., tablets crumble on storage as a kinetically favored polymorph converts to thermodynamically more stable polymorph) or both (e.g., tablets of one polymorph are more susceptible to breakdown at high humidity). Different physical properties of polymorphs can affect their processing. For example, one polymorph might be more likely to form solvates or might be more difficult to filter or wash free of impurities than another due to, for example, the shape or size distribution of particles of it.

[0045] As used herein, the term “solvate” means a compound which further includes a stoichiometric or non-stoichiometric amount of solvent such as water, acetone, ethanol, methanol, dichloromethane, 2-propanol, or the like, bound by non-covalent intermol ecul ar forces.

[0046] As used herein, the term “stable compounds” refers to compounds which possess stability sufficient to allow manufacture and which maintain the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., formulation into therapeutic products, intermediates for use in production of therapeutic compounds, isolatable or storable intermediate compounds, treating a disease or disorder responsive to therapeutic agents).

[0047] As used herein, the term “stereoisomer” refers to both enantiomers and diastereomers. As used herein, the term “substantially free of other stereoisomers” means less than 25% of other stereoisomers, preferably less than 10% of other stereoisomers, more preferably less than 5% of other stereoisomers and most preferably less than 2% of other stereoisomers, or less than "X"% of other stereoisomers (wherein X is a number between 0 and 100, inclusive) are present. Methods of obtaining or synthesizing diastereomers are well known in the art and may be applied as practicable to final compounds or to starting material or intermediates. Other embodiments are those wherein the compound is an isolated compound. The term “at least X% enantiomerically enriched” as used herein means that at least X% of the compound is a single enantiomeric form, wherein X is a number between 0 and 100, inclusive.

[0048] As used herein, the terms “treatment” or “treating” a disease or disorder refers to a method of reducing, delaying or ameliorating such a condition before or after it has occurred. Treatment may be directed at one or more effects or symptoms of a disease and/or the underlying pathology. The treatment can be any reduction and can be, but is not limited to, the complete ablation of the disease or the symptoms of the disease. Treating or treatment thus refers to any indicia of success in the therapy or amelioration of an injury, disease, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving or stabilizing a patient's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters, for example, the results of a physical examination, neuropsychiatric exams, and/or a psychiatric evaluation. As compared with an equivalent untreated control, such reduction or degree of amelioration may be at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100% as measured by any standard technique.

[0049] As used herein, the term “acyl” refers to an alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, heterocyclylcarbonyl, or heteroarylcarbonyl substituent, any of which may be further substituted by substituents.

[0050] As used herein, the terms "alk" or "alkyl" refer to straight or branched chain or cyclic hydrocarbon groups having 1 to 12 carbon atoms, preferably 1 to 8 carbon atoms, containing no unsaturation. The expression "lower alkyl" refers to alkyl groups of 1 to 4 carbon atoms (inclusive). Whenever it appears herein, a numerical range such as " 1 to 10" refers to each integer in the given range; e.g., "1 to 10 carbon atoms" means that the alkyl group can consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although the present definition also covers the occurrence of the term "alkyl" where no numerical range is designated. In some embodiments, “alkyl” can be a C _1-6 alkyl group. In some embodiments, “alkyl” can be a C _1-3 alkyl group. [0051] As used herein, the term "alkenyl" refers to straight or branched chain hydrocarbon groups of 2 to 10, preferably 2 to 4, carbon atoms having at least one double bond. Where an alkenyl group is bonded to a nitrogen atom, it is preferred that such group not be bonded directly through a carbon bearing a double bond.

[0052] As used herein, the term “alkoxy” refers to an -O-alkyl radical.

[0053] As used herein, the term “alkylenedioxo” refers to a divalent species of the structure -O-R- O-, in which R represents an alkylene.

[0054] As used herein, the term "alkynyl" refers to straight or branched chain hydrocarbon groups of 2 to 10, preferably 2 to 4, carbon atoms having at least one triple bond. Where an alkynyl group is bonded to a nitrogen atom, it is preferred that such group not be bonded directly through a carbon bearing a triple bond.

[0055] As used herein, the term "alkylene" refers to a divalent straight chain bridge of 1 to 5 carbon atoms connected by single bonds (e.g., -(CH ₂ ) _X - , wherein x is 1 to 5), which may be substituted with 1 to 3 lower alkyl groups.

[0056] As used herein, the term "alkenylene" refers to a straight chain bridge of 2 to 5 carbon atoms having one or two double bonds that is connected by single bonds and may be substituted with 1 to 3 lower alkyl groups. Exemplary alkenylene groups are -CH=CH-CH=CH-, -CH ₂ -CH=CH-, - CH ₂ -CH=CH-CH ₂ -, -C(CH ₃ ) ₂ CH=CH- and -CH(C ₂ H ₅ )-CH=CH-.

[0057] As used herein, the term "alkynylene" refers to a straight chain bridge of 2 to 5 carbon atoms that has a triple bond therein, is connected by single bonds, and may be substituted with 1 to 3 lower alkyl groups. Exemplary alkynylene groups are -C=C-, -CH ₂ -OC-, -CH(CH ₃ )C=C- and - C=C-CH(C ₂ H ₅ )CH ₂ -.

[0058] As used herein, the term “arylalkyl” refers to a moiety in which an alkyl hydrogen atom is replaced by an aryl group.

[0059] As used herein, the terms “cycloalkyl” and "cycloalkenyl" as employed herein refer to a cyclic alkyl group and includes saturated and partially unsaturated cyclic, respectively, hydrocarbon groups having 3 to 12 carbons, preferably 3 to 8 carbons.

[0060] As used herein, the terms “aromatic”, “ar” or “aryl” refer to a radical with 6 to 14 ring atoms (e.g., C _6-14 aromatic or C _6-14 aryl) that has at least one ring having a conjugated pi electron system which is carbocyclic (e.g., phenyl, fluorenyl, naphthyl, and anthracene). An aryl group may be, for example, 6 membered monocyclic, 10 membered bicyclic or 14 membered tricyclic ring systems, each with 6 to 14 carbon atoms. [0061] As used herein, the term “halo” or "halogen" refers to any radical of fluorine, chlorine, bromine or iodine.

[0062] As used herein, the term "heteroaryl" or, alternatively, "heteroaromatic" refers to a refers to a radical of a 5-18 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic, tetracyclic and the like) aromatic ring system (e.g., having 6, 10 or 14 n electrons shared in a cyclic array) having ring carbon atoms and 1-6 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, phosphorous and sulfur ("5-18 membered heteroaryl"). Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings. Whenever it appears herein, a numerical range such as "5 to 18" refers to each integer in the given range; e.g., "5 to 18 ring atoms" means that the heteroaryl group can consist of 5 ring atoms, 6 ring atoms, etc., up to and including 18 ring atoms. In some instances, a heteroaryl can have 5 to 14 ring atoms. In some embodiments, the heteroaryl has, for example, bivalent radicals derived from univalent heteroaryl radicals whose names end in "-yl" by removal of one hydrogen atom from the atom with the free valence are named by adding "-ene" to the name of the corresponding univalent radical, e.g., a pyridyl group with two points of attachment is a pyridylene. The term “heteroaryl”, for example, may refer to a monocyclic or fused ring (i.e., rings which share an adjacent pair of atoms) group of 5 to 12 ring atoms containing one, two, three or four ring heteroatoms selected from N, O, or S, the remaining ring atoms being C, and, in addition, having a completely conjugated pi-electron system, wherein 0, 1, 2, 3, or 4 atoms of each ring may be substituted by a substituent. Examples, without limitation, of heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, quinazoline, isoquinoline, purine and carbazole.

[0063] As used herein, the terms "heterocycle", "heterocyclic"or "heterocyclo" refer to fully saturated or partially unsaturated cyclic groups, for example, 3 to 7 membered monocyclic, 7 to 12 membered bicyclic, or 10 to 15 membered tricyclic ring systems, which have at least one heteroatom in at least one ring, wherein 0, 1, 2 or 3 atoms of each ring may be substituted by a substituent. Each ring of the heterocyclic group containing a heteroatom may have 1, 2, 3 or 4 heteroatoms selected from nitrogen atoms, oxygen atoms and/or sulfur atoms, where the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quatemized. The heterocyclic group may be attached at any heteroatom or carbon atom of the ring or ring system. [0064] As used herein, the term “heterocyclyl” refers to fully saturated or partially unsaturated cyclic groups, for example, 3- to 7- membered monocyclic, 7- to 12- membered bicyclic, or 10- to 15- membered tricyclic ring systems, which have at least one heteroatom in at least one ring, wherein 0, 1, 2 or 3 atoms of each ring may be substituted by a substituent. Each ring of the heterocyclyl group containing a heteroatom may have 1, 2, 3 or 4 heteroatoms selected from nitrogen atoms, oxygen atoms and/or sulfur atoms, where the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quatemized. The heterocyclyl group may be attached at any heteroatom or carbon atom of the ring or ring system.

[0065] As used herein, the term “oxo” refers to an oxygen atom, which forms a carbonyl when attached to carbon, an N-oxide when attached to nitrogen, and a sulfoxide or sulfone when attached to sulfur.

[0066] As used herein, the term “substituents” refers to a group “substituted” on any functional group delineated herein, e.g., alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, or heteroaryl group at any atom of that group. Suitable substituents include, without limitation halogen, CN, NO2, OR ¹⁵ , SR ¹⁵ , S(O) ₂ OR ¹⁵ , NR ¹⁵ R ¹⁶ , C ₁ -C ₂ perfluoroalkyl, C ₁ -C ₂ perfluoroalkoxy, 1,2- methylenedioxy, C(O)OR ¹⁵ , C(O)NR ¹⁵ R ¹⁶ , OC(O)NR ¹⁵ R ¹⁶ , NR ¹⁵ C(O)NR ¹⁵ R ¹⁶ , C(NR ¹⁶ )NR ¹⁵ R ¹⁶ , NR ¹⁵ C(NR ¹⁶ )NR ¹⁵ R ¹⁶ , S(O) ₂ NR ¹⁵ R ¹⁶ , R ¹⁷ , C(O)R ¹⁷ , NR ¹⁵ C(O)R ¹⁷ , S(O)R ¹⁷ , S(O) ₂ R ¹⁷ , R ¹⁶ , OXO, C(O)R ¹⁶ , C(O)(CH ₂ )nOH, (CH ₂ )nOR ¹⁵ , (CH ₂ )nC(O)NR ¹⁵ R ¹⁶ , NR ¹⁵ S(O) ₂ R ¹⁷ , where n is independently 0-6 inclusive. Each R ¹⁵ is independently hydrogen, C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl. Each R ¹⁶ is independently hydrogen, alkenyl, alkynyl, C ₃ -C ₆ cycloalkyl, aryl, heterocyclyl, heteroaryl, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkyl substituted with C ₃ -C ₆ cycloalkyl, aryl, heterocyclyl or heteroaryl. Each R ¹⁷ is independently C ₃ -C ₆ cycloalkyl, aryl, heterocyclyl, heteroaryl, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkyl substituted with C ₃ -C ₆ cycloalkyl, aryl, heterocyclyl or heteroaryl. Each Cs-Ce cycloalkyl, aryl, heterocyclyl, heteroaryl and C ₁ -C ₄ alkyl in each R ¹⁵ , R ¹⁶ and R ¹⁷ can optionally be substituted with halogen, CN, C ₁ -C ₄ alkyl, OH, C ₁ -C ₄ alkoxy, NH ₂ , C ₁ -C ₄ alkylamino, C ₁ -C ₄ dialkylamino, C ₁ -C ₂ perfluoroalkyl, C ₁ -C ₂ perfluoroalkoxy, or 1,2-methylenedioxy.

[0067] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0068] The compounds of this invention may contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, single enantiomers, individual diastereomers and diastereomeric mixtures. All such isomeric forms of these compounds are expressly included in the present invention. The compounds of this invention may also be represented in multiple tautomeric forms, in such instances, the invention expressly includes all tautomeric forms of the compounds described herein. All such isomeric forms of such compounds are expressly included in the present invention. All crystal forms of the compounds described herein are expressly included in the present invention.

Detailed Description of the Invention

[0069] The invention provides novel sulfonanilide and benzylsulfonyl derivatives, including salts, solvates, hydrates and polymorphs thereof, as TRPML modulators. The invention also provides pharmaceutical compositions comprising a compound of this invention and the use of such compositions in treating a range of diseases and conditions associated with TRPML or related to TRPML activities, such as lysosome storage diseases, muscular dystrophy, age-related common neurodegenerative diseases, ROS or oxidative stress related diseases, and damages caused in skin or photoaging.

[0070] In one aspect, the invention generally relates to a compound having the structural formula I: or a pharmaceutically acceptable form or an isotope derivative thereof, wherein:

R ¹ is a 5-membered heteroaryl substituted with 1, 2, or 3 substituents at least one of which is selected from the group consisting of C _1-5 alkyl, CF ₃ , CH ₂ CF ₃ , CN, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR’; and wherein any alkyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, halo, OH, OC _1-3 alkyl, and CN;

R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8-membered) heterocyclyl which optionally comprises a further heteroatom selected from the group consisting of nitrogen optionally substituted with alkyl or aryl, oxygen and S(O) _n , wherein said 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8-membered) heterocyclyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8- membered) heterocyclyl spiro fused to a C3.7 cycloalkyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8- membered) bridged heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; each of R ⁴ and R ⁵ is independently H, C _1-3 alkyl, halogen, oxo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, or S(O) ₂ NRR’; each of R and R’ is independently H, or C _1-5 alkyl or, optionally, R and R’, together with the nitrogen to which they are attached to, form a 4- to 6-membered (e.g., 4-, 5- or 6-membered) ring;

X is CR ⁶ R ⁷ , or NR ⁶ , wherein each of R ⁶ and R ⁷ is independently H, or C1-C3 alkyl; and n is 0, 1 or 2.

[0071] In certain embodiments, R ¹ is a 5-membered heteroaryl substituted with S(O) ₂ NRR’, wherein each of R and R’ is C _1-3 alkyl (e.g., Ci, C ₂ or C3 alkyl).

[0072] In certain embodiments, R ¹ is a 5-membered heteroaryl substituted with S(O) ₂ NRR’, wherein R and R’, together with the nitrogen to which they are attached to, form a 4- to 6-membered (e.g., 4-, 5- or 6-membered) ring.

[0073] In certain embodiments, the 5-membered heteroaryl comprises the heteroatom S.

[0074] In certain embodiments, R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 6-membered heterocyclyl optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR. [0075] In certain embodiments, R ¹ is a 5-membered heteroaryl substituted with S(O) ₂ R, wherein R is a linear, branched or cyclic C _1-5 alkyl (e.g., a linear or branched C _1-5 alkyl or a cyclic C _3-5 alkyl).

[0076] In certain embodiments, R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 6-membered heterocyclyl substituted with 2 C _1-3 alkyl (e.g., C ₁ , C ₂ or C ₃ alkyl) groups.

[0077] In certain embodiments, R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 5-membered heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR.

[0078] In certain embodiments, R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 4-membered heterocyclyl optionally substituted with C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, or S(O) ₂ NRR.

[0079] In certain embodiments, R ⁵ is H.

[0080] In certain embodiments, R ¹ is selected from the group consisting of:

wherein a dotted line indicates a point of attachment.

[0081] In certain embodiments, NR ² R ³ is selected from the group consisting of:

wherein a dotted line indicates a point of attachment.

[0082] In certain embodiments, the compound of the invention has the structural formula Ila:

Ila wherein

Z is CR ^a R ^b , NR ^C or SO ₂ ;

R ⁴ is H, C _1-3 alkyl or halogen; each of R ^a , R ^b and R ^c is independent H, C _1-3 alkyl or halogen, or two of R ^a , R ^b and R ^c together with the carbon or hetero atoms to which they are attached to, are linked to form a 3- to 7-membered ring. [0083] In certain embodiments, the compound of the invention has the structural formula lib: wherein

R ⁴ is H, C _1-3 alkyl or halogen; each of R ^a , R ^b and R ^c is independent H, C _1-3 alkyl or halogen, or two of R ^a , R ^b and R ^c together with the carbon or hetero atoms to which they are attached to, are linked to form a 3- to 7-membered ring.

[0084] In certain embodiments, the compound of the invention has the structural formula lIc: wherein

R ⁴ is H, C _1-3 alkyl or halogen; each of R ^a , R ^b and R ^c is independent H, C _1-3 alkyl or halogen, or two of R ^a , R ^b and R ^c together with the carbon or hetero atoms to which they are attached to, are linked to form a 3- to 7-membered ring.

[0085] In certain embodiments, the compound of the invention has the structural formula lid: wherein

R ⁴ is H, C _1-3 alkyl or halogen; each of R ^a , R ^b and R ^c is independent H, C _1-3 alkyl or halogen, or two of R ^a , R ^b and R ^c together with the carbon or hetero atoms to which they are attached to, are linked to form a 3- to 7-membered ring.

[0086] In certain embodiments, each of R ^a and R ^b is H.

[0087] In certain embodiments, each of R ^a and R ^b is a halogen atom.

[0088] In certain embodiments, each of R ^a and R ^b is methyl.

[0089] In certain embodiments, each of R ^a and R ^b is ethyl.

[0090] In certain embodiments, R ^a is methyl and R ^b is ethyl.

[0091] In certain embodiments, R ^a and R ^b together with the carbon to which they are attached to, are linked to form cyclopropyl.

[0092] In certain embodiments, each of R and R’ is C _1-3 alkyl.

[0093] In certain embodiments, each of R and R’ is methyl.

[0094] In certain embodiments, R and R’, together with the nitrogen to which they are attached to, form a 4- to 6-membered ring optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, halo, OH, OC _1-3 alkyl, and CN. [0095] In certain embodiments, R ⁴ is F or Cl.

[0096] In certain embodiments, R ⁴ is C _1-3 alkyl.

[0097] In certain embodiments, R ⁴ is methyl.

[0098] Exemplary compounds of the invention include but not limited to:

[0099] In certain embodiments, a compound of the invention has one or more (e.g., 1, 2, 3) deuterium atoms replacing one or more (e.g., 1, 2, 3) hydrogen atoms. In certain embodiments, a compound of the invention has one deuterium atom replacing one hydrogen atom.

[00100] In another aspect, the invention generally relates to a pharmaceutical composition comprising a compound disclosed herein, effective to treat or reduce one or more diseases or disorders, in a mammal, including a human, and a pharmaceutically acceptable excipient, carrier, or diluent.

[00101] In yet another aspect, the invention generally relates to a pharmaceutical composition comprising an amount of a compound of formula I: or a pharmaceutically acceptable form or an isotope derivative thereof, wherein:

R ¹ is a 5-membered heteroaryl substituted with 1, 2, or 3 substituents at least one of which is selected from the group consisting of C _1-5 alkyl, CF ₃ , CH ₂ CF ₃ , CN, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR’; and wherein any alkyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, halo, OH, OC _1-3 alkyl, and CN;

R ² and R ³ , together with the nitrogen to which they are attached to, are linked to form a 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8-membered) heterocyclyl which optionally comprises a further heteroatom selected from the group consisting of nitrogen optionally substituted with alkyl or aryl, oxygen and S(O) _n , wherein said 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8-membered) heterocyclyl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, and S(O) ₂ NRR; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8- membered) heterocyclyl spiro fused to a C3.7 cycloalkyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; or R ² and R ³ , together with the nitrogen to which they are attached to are linked to form a 4- to 8-membered (e.g., 4-, 5-, 6-, 7, or 8- membered) bridged heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, and C(=O)NRR’; and wherein any alkyl or aryl is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of C _1-3 alkyl, aryl, halo, OH and CN; each of R ⁴ and R ⁵ is independently H, C _1-3 alkyl, halogen, oxo, OH, CN, OR, NHR, NRR’, N(R)C(=O)R’, N(R)C(=O)(O)R’, OC(=O)NRR’, C(=O)R, C(=O)NRR’, N(R)S(O) ₂ R’, S(O) ₂ R, or S(O) ₂ NRR’; each of R and R’ is independently H, or C _1-5 alkyl or, optionally, R and R’, together with the nitrogen to which they are attached to, form a 4- to 6-membered (e.g., 4-, 5- or 6-membered) ring;

X is CR ⁶ R ⁷ , or NR ⁶ , wherein each of R ⁶ and R ⁷ is independently H, or C1-C3 alkyl; and n is 0, 1 or 2, effective to treat, or reduce one or more diseases or disorders, in a mammal, including a human, and a pharmaceutically acceptable excipient, carrier, or diluent.

[00102] In certain embodiments, the pharmaceutical composition is suitable for oral administration. In certain embodiments, the pharmaceutical composition is suitable for topital administration.

[00103] In yet another aspect, the invention generally relates to a unit dosage form comprising a pharmaceutical composition disclosed herein.

[00104] In certain embodiments, the unit dosage form is a tablet or a capsule.

[00105] In yet another aspect, the invention generally relates to a method for treating or reducing a disease or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

[00106] In certain embodiments, the disease or disorder is mediated by loss-of-function in TRPML1, including ML4 and NPC.

[00107] In certain embodiments, the disease or disorder is a lysosome storage disease, or a related disease or disorder.

[00108] In certain embodiments, the disease or disorder is selected from the group consisting of age-related neurodegenerative disease, including Alzheimer’s Disease, Parkinson’s Disease, and Huntington’s Disease, or a related disease or disorder. [00109] In certain embodiments, the disease or disorder is muscular dystrophy, or a related disease or disorder.

[00110] In certain embodiments, the disease or disorder is oxidative stress or ROS, or a related disease or disorder.

[00111] In yet another aspect, the invention generally relates to a method for treating or reducing the effect of aging comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

[00112] In certain embodiments, the effect of aging comprises skin aging.

[00113] In certain embodiments, the effect of aging comprises photoaging.

[00114] In yet another aspect, the invention generally relates to a method for treating or reducing oxidative stress or ROS related diseases or disorder, comprising administering to a subject in need thereof an effective amount of a TRPML1 agonist or a composition comprising of a TRPML1 agonist.

[00115] In yet another aspect, the invention generally relates to a method for treating or reducing oxidative stress or ROS related diseases or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

[00116] In certain embodiments, administration is via oral administration.

[00117] In certain embodiments, administration is via topical administration.

[00118] In yet another aspect, the invention generally relates to use of a compound disclosed herein, and a pharmaceutically acceptable excipient, carrier, or diluent, in preparation of a medicament for treating a disease or disorder.

[00119] In certain embodiments, the disease or disorder is selected from the group consisting of age-related neurodegenerative disease, including Alzheimer’s Disease, Parkinson’s Disease, and Huntington’s Disease, or a related disease or disorder.

[00120] In certain embodiments, the disease or disorder is muscular dystrophy, or a related disease or disorder.

[00121] In certain embodiments, the disease or disorder is oxidative stress or ROS, or a related disease or disorder.

[00122] In certain embodiments, the disease or disorder is skin aging or photoaging.

[00123] In certain embodiments, the compound is any of those shown in Table 1.

[00124] The specific approaches and compounds disclosed herein are not intended to be limiting. The chemical structures in the schemes herein depict variables that are hereby defined commensurately with chemical group definitions (moieties, atoms, etc.) of the corresponding position in the compound formulae herein, whether identified by the same variable name (e.g., R ¹ , R ² , R, R', X, etc.) or not. The suitability of a chemical group in a compound structure for use in synthesis of another compound structure is within the knowledge of one of ordinary skill in the art. Additional methods of synthesizing compounds of the formulae herein and their synthetic precursors, including those within routes not explicitly shown in schemes herein, are within the means of chemists of ordinary skill in the art. Methods for optimizing reaction conditions, if necessary minimizing competing by-products, are known in the art. The methods described herein may also additionally include steps, either before or after the steps described specifically herein, to add or remove suitable protecting groups in order to ultimately allow synthesis of the compounds herein. In addition, various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing the applicable compounds are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3 ^rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser ’s Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions thereof.

[00125] The methods delineated herein contemplate converting compounds of one formula to compounds of another formula. The process of converting refers to one or more chemical transformations, which can be performed in situ, or with isolation of intermediate compounds. The transformations can include reacting the starting compounds or intermediates with additional reagents using techniques and protocols known in the art, including those in the references cited herein. Intermediates can be used with or without purification (e.g., filtration, distillation, sublimation, crystallization, trituration, solid phase extraction, and chromatography).

[00126] Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds.

[00127] Certain compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and /raw.s-isomers, atropisomers, R- and 5-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.

[00128] Isomeric mixtures containing any of a variety of isomer ratios may be utilized in accordance with the present invention. For example, where only two isomers are combined, mixtures containing 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0 isomer ratios are contemplated by the present invention. Those of ordinary skill in the art will readily appreciate that analogous ratios are contemplated for more complex isomer mixtures.

[00129] If, for instance, a particular enantiomer of a compound of the present invention is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic methods well known in the art, and subsequent recovery of the pure enantiomers.

[00130] Solvates and polymorphs of the compounds of the invention are also contemplated herein. Solvates of the compounds of the present invention include, for example, hydrates.

[00131] The invention also provides compositions comprising an effective amount of a compound of any of the formulae herein, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph or prodrug, if applicable, of said compound; and an acceptable carrier. Preferably, a composition of this invention is formulated for pharmaceutical use (“a pharmaceutical composition”), wherein the carrier is a pharmaceutically acceptable carrier. The carrier(s) must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and, in the case of a pharmaceutically acceptable carrier, not deleterious to the recipient thereof in amounts typically used in medicaments.

[00132] Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. [00133] The pharmaceutical compositions of the invention include those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. In certain embodiments, the compound of the formulae herein is administered transdermally (e.g., using a transdermal patch). Other formulations may conveniently be presented in unit dosage form, e.g., tablets and sustained release capsules, and in liposomes, and may be prepared by any methods well known in the art of pharmacy. See, for example, Remington’s Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA (17th ed. 1985).

[00134] Such preparative methods include the step of bringing into association with the molecule to be administered ingredients such as the carrier that constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers, liposomes or finely divided solid carriers or both, and then if necessary shaping the product.

[00135] In certain preferred embodiments, the compound is administered orally. Compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, sachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion, or packed in liposomes and as a bolus, etc. Soft gelatin capsules can be useful for containing such suspensions, which may beneficially increase the rate of compound absorption.

[00136] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface- active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets optionally may be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein. Methods of formulating such slow or controlled release compositions of pharmaceutically active ingredients, such as those herein and other compounds known in the art, are known in the art and described in several issued US Patents, some of which include, but are not limited to, US Patent Nos. 4,369,172; and 4,842,866, and references cited therein. Coatings can be used for delivery of compounds to the intestine (see, e.g., U.S. Patent Nos. 6,638,534, 5,217,720, and 6,569,457, 6,461,631, 6,528,080, 6,800,663, and references cited therein). A useful formulation for the compounds of this invention is the form of enteric pellets of which the enteric layer comprises hydroxypropylmethylcellulose acetate succinate. [00137] In the case of tablets for oral use, carriers that are commonly used include lactose and com starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.

[00138] Compositions suitable for topical administration include lozenges comprising the ingredients in a flavored basis, usually sucrose and acacia or tragacanth; and pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia.

[00139] Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and nonaqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets. [00140] Such injection solutions may be in the form, for example, of a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3 -butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant. [00141] The pharmaceutical compositions of this invention may be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.

[00142] The pharmaceutical compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.

[00143] Topical administration of the pharmaceutical compositions of this invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application. For application topically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topically-transdermal patches and iontophoretic administration are also included in this invention.

[00144] Particularly favored derivatives and prodrugs are those that increase the bioavailability of the compounds of this invention when such compounds are administered to a mammal (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment (e.g., the brain or central nervous system) relative to the parent species. Preferred prodrugs include derivatives where a group that enhances aqueous solubility or active transport through the gut membrane is appended to the structure of formulae described herein. (See, e.g., Alexander, et al. 1988 J Med Chem 31, 318-322; Bundgaard 1985 Elsevier: Amsterdam 1-92; Bundgaard, et al. 1987 J Med Chem 30, 451-454; Bundgaard, H. A Textbook of Drug Design and Development, Harwood Academic Publ.: Switzerland, 1991, 113-191; Digenis, et al. Handbook of Experimental Pharmacology 1975 28, 86- 112; Friis, et al. A Textbook of Drug Design and Development, 2 ed.; Overseas Publ.: Amsterdam, 1996, 351-385; Pitman 1981 Med Res Rev 1, 189-214.)

[00145] Application of the subject therapeutics may be local, so as to be administered at the site of interest. Various techniques can be used for providing the subject compositions at the site of interest, such as injection, use of catheters, trocars, projectiles, pluronic gel, stents, sustained drug release polymers or other device which provides for internal access.

[00146] According to another embodiment, the invention provides a method of impregnating an implantable drug release device comprising the step of contacting said drug release device with a compound or composition of this invention. Implantable drug release devices include, but are not limited to, biodegradable polymer capsules or bullets, non-degradable, diffusible polymer capsules and biodegradable polymer wafers.

[00147] According to another embodiment, the invention provides an implantable medical device coated with a compound or a composition comprising a compound of this invention, such that said compound is therapeutically active.

[00148] In another embodiment, a composition of the present invention further comprises a second therapeutic agent. The second therapeutic agent includes any compound or therapeutic agent known to have or that demonstrates advantageous properties when administered alone or with a compound of any of the formulae herein. Drugs that could be usefully combined with these compounds include other kinase inhibitors and/or other chemotherapeutic agents for the treatment of the diseases and disorders discussed above.

[00149] Such agents are described in detail in the art. Preferably, the second therapeutic agent is an agent useful in the treatment or prevention of cancer.

[00150] Even more preferably the second therapeutic agent co-formulated with a compound of this invention is an agent useful in the treatment of TRPML mediated disease/disorders.

[00151] In another embodiment, the invention provides separate dosage forms of a compound of this invention and a second therapeutic agent that are associated with one another. The term “associated with one another” as used herein means that the separate dosage forms are packaged together or otherwise attached to one another such that it is readily apparent that the separate dosage forms are intended to be sold and administered together (within less than 24 hours of one another, consecutively or simultaneously). [00152] In the pharmaceutical compositions of the invention, the compound of the present invention is present in an effective amount. As used herein, the term “effective amount” refers to an amount which, when administered in a proper dosing regimen, is sufficient to reduce or ameliorate the severity, duration or progression of the disorder being treated, prevent the advancement of the disorder being treated, cause the regression of the disorder being treated, or enhance or improve the prophylactic or therapeutic effect(s) of another therapy.

[00153] The interrelationship of dosages for animals and humans (based on milligrams per meter squared of body surface) is described in Freireich, et al. 1966 Cancer Chemother Rep 50: 219. Body surface area may be approximately determined from height and weight of the patient. (See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardley, N.Y., 1970, 537.) An effective amount of a compound of this invention can range from about 0.001 mg/kg to about 500 mg/kg, more preferably 0.01 mg/kg to about 50 mg/kg, more preferably 0.1 mg/kg to about 2.5 mg/kg. Effective doses will also vary, as recognized by those skilled in the art, depending on the diseases treated, the severity of the disease, the route of administration, the sex, age and general health condition of the patient, excipient usage, the possibility of co-usage with other therapeutic treatments such as use of other agents and the judgment of the treating physician.

[00154] For pharmaceutical compositions that comprise a second therapeutic agent, an effective amount of the second therapeutic agent is between about 20% and 100% of the dosage normally utilized in a monotherapy regime using just that agent. Preferably, an effective amount is between about 70% and 100% of the normal monotherapeutic dose. The normal monotherapeutic dosages of these second therapeutic agents are well known in the art. (See, e.g., Wells, et al., eds. 2000 Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, Conn. ; PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. 2000, each of which references are entirely incorporated herein by reference.

[00155] It is expected that some of the second therapeutic agents referenced above will act synergistically with the compounds of this invention. When this occurs, its will allow the effective dosage of the second therapeutic agent and/or the compound of this invention to be reduced from that required in a monotherapy. This has the advantage of minimizing toxic side effects of either the second therapeutic agent of a compound of this invention, synergistic improvements in efficacy, improved ease of administration or use and/or reduced overall expense of compound preparation or formulation. [00156] The invention also provides a method of treating a subject suffering from or susceptible to a disease or disorder or symptom thereof (e.g., those delineated herein) comprising the step of administering to said subject an effective amount of a compound or a composition of this invention. Some diseases are well known in the art and are also disclosed herein.

[00157] In certain embodiments, the methods disclosed herein are suitable for treating diseases or disorders that are mediated by the TRPMLs. In certain embodiments, the methods disclosed herein are suitable for treating disease or disorders that are mediated by loss-of-function in TRPML1, including ML4 and NPC.

[00158] In certain embodiments, the disease is one of the lysosomal storage diseases, such as Niemen-Pick C (NPC) disease.

[00159] In certain embodiments, the methods disclosed herein are suitable for treating diseases or disorders that are age-related including common neurodegenerative diseases, such as AD, PD, and HD.

[00160] In certain embodiments, the methods disclosed herein are suitable for treating type IV Mucolipidosis (ML4), a neurodegenerative LSD caused by human mutations in TRPML1.

[00161] In certain embodiments, the methods disclosed herein are suitable for treating a ROS or oxidative stress related disease or disorder.

[00162] In certain embodiments, the methods disclosed herein are suitable for treating diseases or disorders due or related to ageing.

[00163] The term “co-administered” as used herein means that the second therapeutic agent may be administered together with a compound of this invention as part of a single dosage form (such as a composition of this invention comprising a compound of the invention and an second therapeutic agent as described above) or as separate, multiple dosage forms. Alternatively, the additional agent may be administered prior to, consecutively with, or following the administration of a compound of this invention. In such combination therapy treatment, both the compounds of this invention and the second therapeutic agent(s) are administered by conventional methods. The administration of a composition of this invention comprising both a compound of the invention and a second therapeutic agent to a subject does not preclude the separate administration of that same therapeutic agent, any other second therapeutic agent or any compound of this invention to said subject at another time during a course of treatment.

[00164] Effective amounts of these second therapeutic agents are well known to those skilled in the art and guidance for dosing may be found in patents and published patent applications referenced herein, as well as in Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, Conn. (2000); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. (2000), and other medical texts. However, it is well within the skilled artisan’s purview to determine the second therapeutic agent’s optimal effective-amount range.

[00165] In one embodiment of the invention where a second therapeutic agent is administered to a subject, the effective amount of the compound of this invention is less than its effective amount would be where the second therapeutic agent is not administered. In another embodiment, the effective amount of the second therapeutic agent is less than its effective amount would be where the compound of this invention is not administered. In this way, undesired side effects associated with high doses of either agent may be minimized. Other potential advantages (including without limitation improved dosing regimens and/or reduced drug cost) will be apparent to those of skill in the art.

[00166] In yet another aspect, the invention provides the use of a compound of any of the formulae herein alone or together with one or more of the above-described second therapeutic agents in the manufacture of a medicament, either as a single composition or as separate dosage forms, for treatment or prevention in a subject of a disease, disorder or symptom set forth above. Another aspect of the invention is a compound of the formulae herein for use in the treatment or prevention in a subject of a disease, disorder or symptom thereof delineated herein.

[00167] In other aspects, the methods herein include those further comprising monitoring subject response to the treatment administrations. Such monitoring may include periodic sampling of subject tissue, fluids, specimens, cells, proteins, chemical markers, genetic materials, etc. as markers or indicators of the treatment regimen. In other methods, the subject is prescreened or identified as in need of such treatment by assessment for a relevant marker or indicator of suitability for such treatment.

[00168] In one embodiment, the invention provides a method of monitoring treatment progress. The method includes the step of determining a level of diagnostic marker (Marker) (e.g., any target or cell type delineated herein modulated by a compound herein) or diagnostic measurement (e.g., screen, assay) in a subject suffering from or susceptible to a disorder or symptoms thereof delineated herein, in which the subject has been administered a therapeutic amount of a compound herein sufficient to treat the disease or symptoms thereof. The level of Marker determined in the method can be compared to known levels of Marker in either healthy normal controls or in other afflicted patients to establish the subject’s disease status. In preferred embodiments, a second level of Marker in the subject is determined at a time point later than the determination of the first level, and the two levels are compared to monitor the course of disease or the efficacy of the therapy. In certain preferred embodiments, a pre-treatment level of Marker in the subject is determined prior to beginning treatment according to this invention; this pre-treatment level of Marker can then be compared to the level of Marker in the subject after the treatment commences, to determine the efficacy of the treatment.

[00169] In certain method embodiments, a level of Marker or Marker activity in a subject is determined at least once. Comparison of Marker levels, e.g., to another measurement of Marker level obtained previously or subsequently from the same patient, another patient, or a normal subject, may be useful in determining whether therapy according to the invention is having the desired effect, and thereby permitting adjustment of dosage levels as appropriate. Determination of Marker levels may be performed using any suitable sampling/expression assay method known in the art or described herein. Preferably, a tissue or fluid sample is first removed from a subject. Examples of suitable samples include blood, urine, tissue, mouth or cheek cells, and hair samples containing roots. Other suitable samples would be known to the person skilled in the art. Determination of protein levels and/or mRNA levels (e.g., Marker levels) in the sample can be performed using any suitable technique known in the art, including, but not limited to, enzyme immunoassay, ELISA, radiolabeling/assay techniques, blotting/chemiluminescence methods, real-time PCR, and the like. [00170] The present invention also provides kits for use to treat diseases, disorders, or symptoms thereof, including those delineated herein. These kits comprise: a) a pharmaceutical composition comprising a compound of any of the formula herein or a salt thereof; or a prodrug, or a salt of a prodrug thereof; or a hydrate, solvate, or polymorph thereof, wherein said pharmaceutical composition is in a container; and b) instructions describing a method of using the pharmaceutical composition to treat the disease, disorder, or symptoms thereof, including those delineated herein.

[00171] The container may be any vessel or other sealed or sealable apparatus that can hold said pharmaceutical composition. Examples include bottles, divided or multi-chambered holders or bottles, wherein each division or chamber comprises a single dose of said composition, a divided foil packet wherein each division comprises a single dose of said composition, or a dispenser that dispenses single doses of said composition. The container can be in any conventional shape or form as known in the art which is made of a pharmaceutically acceptable material, for example a paper or cardboard box, a glass or plastic bottle or jar, a re-sealable bag (for example, to hold a "refill" of tablets for placement into a different container), or a blister pack with individual doses for pressing out of the pack according to a therapeutic schedule. The container employed can depend on the exact dosage form involved, for example a conventional cardboard box would not generally be used to hold a liquid suspension. It is feasible that more than one container can be used together in a single package to market a single dosage form. For example, tablets may be contained in a bottle, which is in turn contained within a box. Preferably, the container is a blister pack.

[00172] The kit may additionally comprise information and/or instructions for the physician, pharmacist or subject. Such memory aids include numbers printed on each chamber or division containing a dosage that corresponds with the days of the regimen which the tablets or capsules so specified should be ingested, or days of the week printed on each chamber or division, or a card which contains the same type of information.

[00173] The following examples are meant to be illustrative of the practice of the invention and not limiting in any way.

Examples

[00174] The structures depicted herein, including the Table 1 structures, may contain certain - NH-, -NH2 (amino) and -OH (hydroxyl) groups where the corresponding hydrogen atom(s) do not explicitly appear; however, they are to be read as -NH-, -NH2 or -OH as the case may be. In certain structures, a stick bond is drawn and is meant to depict a methyl group.

Table 1

*EC50 of Ca ²⁺ Imaging: “+++” ≤ 1 uM; “++” > 1 uM to ≤ 10 uM; “+” > 10 uM;

**EC50 of TFEB activation: “+++” ≤ 1 uM; “++” > 1 uM to < 10 uM; “+” > 10 uM;

N/A: Not available Example 4: Synthesis of N2, N2-dimethyl-N5-[2-(l-piperidyl) phenyl] thiophene-2, 5- disulfonamide

^4E 4F

[00175] Step 1. To a 250 mL three necked flask was added100mL dichloromethane and 2- thiophenesulfonyl chloride (11.5 g, 62.9 mmol, 1.0 eq), then dimethylamine solution in water (21.2 g, 188.8 mmol, 3.0 eq) was added to the mixture, then the reaction mixture was stirred overnight at room temperature, TLC (PE: ethyl acetate=5:l, Rf=0.4) showed the reaction was completed. The aqueous phase was extracted with di chloromethane (100 mL*2). The organic layer was washed with brine (100 mL*l) and dried over Na ₂ SO ₄ , filtered and concentrated under vacuum to afford the compound 4A (9.4g, HPLC:92%) as a gray-white solid, Yield: 78.3%.

[00176] Step 2'. To a 500 mL three necked flask was added100mL CHCL and compound 4A (5.73 g, 30 mmol, 1.0 eq), then Br2 (9.6 g, 60 mmol, 2.0 eq) was added dropwise to the mixture, the reaction mixture was reflux for 48h, TLC (PE: ethyl acetate=5:l, Rf=0.45) showed the reaction was having a new point. The mixture was cooled and concentrated under vacuum and the resulting compound was purified by column chromatography on silica gel to afford the title compound 4B (3.0 g, HPLC: 95%) as a light-yellow solid. Yield: 37.5%.

[00177] Step 3: To a 100 mL round-bottom flask was added 20mL toluene, compound 4B (1.4 g, 5.2 mmol, 1.0 eq), benzyl mercaptan (0.8 g, 5.7 mmol, 1.1 eq), N,N-diisopropylethylamine (1.4 g, 10.4 mmol, 2.0 eq), Pd(dppf)C12 (114 mg, 0.13 mmol, 0.025 eq), 4,5-Bis(diphenylphosphino)-9,9- dimethylxanthene (320 mg, 0.52 mmol, 0.1 eq), the mixture was heat to reflux in the nitrogen atmosphere overnight, TLC (PE: ethyl acetate=5:l, Rf=0.4) showed the reaction was completed and the mixture was cooled and concentrated under vacuum and the resulting compound was purified by column chromatography on silica gel to afford the title compound 4C (1.2 g, HPLC: 88%) as an oil. Yield: 75%.

[00178] Step 4. To a 100 mL three necked flask was added 20mL glacial acetic acid, 5mL water and compound 4C (0.8 g, 2.6 mol, 1.0 eq), then N-chlorosuccinimide (1.84 g, 7.7 mmol, 3.0 eq) was added to the reaction mixture under 10-15 °C, then the reaction mixture was stirred for 1 h at 10-15 °C, TLC (PE: ethyl acetate=5:l, Rf=0.3) showed the reaction was completed. The mixture was concentrated under vacuum and the resulting compound was purified by column chromatography on silica gel to afford the title compound 4D (350 mg, HPLC: 95%) as an off-white solid. Yield: 46.6%. [00179] Step 5: To a 500 mL round-bottom flask was added 1,4-dioxane 100 mL, l-fluoro-2- nitrobenzene (20 g, 0.235 mol, 1.0 eq), potassium carbonate (30 g, 0.587 mol, 2.5 eq) and piperidine (30 g, 0.2114 mol, 0.9 eq) was dropped into the stirred mixture and refluxed for 3h. This reaction was added water 200 mL and extracted with ethyl acetate (50 mL*3). The organic solution was dried over sodium sulfate, filtered and concentrated to afford the title compound 4E (32 g). Yield: 66.8%.

[00180] Step 6: To a 500mL round-bottom flask was added methanol 200 mL, compound 4E (32 g, 0.156 mol, 1.0 eq), Raney Ni catalyst (3 g) was added into the stirred mixture at room temperature overnight. The solution was filtered and concentrated to afford the title compound 4F (10 g, HPLC: 94%). Yield: 36%.

[00181] Step 7: To a 50 mL round-bottom flask was added 4 mL pyridine, compound 4D (92 mg, 0.52 mmol, 1.0 eq) and compound 4F (180 mg, 0.62 mmol, 1.3 eq), the reaction was stirred at 20 °C overnight. TLC (PE: ethyl acetate=3:l, Rf=0.2) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 4 (40 mg, HPLC: 97.3%) as a red solid. Yield: 17.9%. MS-ESI: [M+l] ⁺ = 430.5. ¹ H NMR (400 MHz, DMSO): δ 9.42 (s, 1H), 7.70 - 7.61 (m, 2H), 7.34 (d, 1H), 7.21 - 7.16 (m, 2H), 7.15 - 7.09 (m, 1H), 2.66 (s, 6H), 2.56 - 2.51 (m, 4H), 1.59-1.54 (m, 4H), 1.48-1.42 (m, 2H).

Example 5: Synthesis of N2-[3-chloro-2-(l-piperidyl)phenyl]-N5,N5-dimethylthiophene- 2,5- disulfonamide [00182] Step 1. To a 500 mL round-bottom flask was added 1,4-dioxane 100 mL, l-chloro-2- fluoro-3 -nitrobenzene (10 g, 0.142 mol, 0.9 eq) and potassium carbonate (22 g, 0.4 mol, 2.5 eq). Piperidine (5.6 g, 0.158 mol, 1.0 eq) was dropped into the stirred mixture and refluxed for 3 h. This reaction was added water 50mL and extracted with ethyl acetate (30 mL*3). The organic solution was dried over sodium sulfate, filtered and concentrated to afford the title compound 5A (10 g). Yield: 30%.

[00183] Step 2'. To a 250 mL round-bottom flask was added methanol 100 mL, compound 5A (10 g), Raney Ni catalyst (3 g, 30%wt) was added to the stirred mixture at room temperature overnight. The solution was filtered and concentrated to afford the title compound 5B (4.3 g). Yield: 49%

[00184] Step 3. To a 25 mL round-bottom flask was added pyridine 5 mL, compound 5B (60 mg, 0.287 mmol, 1.0 eq) and compound 4D (100 mg, 0.347 mmol, 1.3 eq) was dropped into the stirred mixture at room temperature for 2 h. The solution was concentrated and subjected to silica gel to afford the title compound 5 (20 mg, HPLC: 99%). Yield: 15% Overall Yield: 2.25%.

MS-ESI: [M+l] ⁺ = 463.8. ¹ HNMR (400 MHz, DMSO): 5 9.52 (s, 1H), 7.72 - 7.66 (m, 2H), 7.30 - 7.24 (m, 2H), 7.22 - 7.17 (m, 1H), 3.28-3.10 (m, 2H), 2.66 (s, 6H), 2.46 - 2.37 (m, 2H), 1.7 - 1.5 (m, 6H).

Example 6: Synthesis of N2-[3,5-difluoro-2-(l-piperidyl)phenyl]-N5,N5-dimethylthioph ene- 2,5- disulfonamide

[00185] Step 1. To a 100 mL three necked flask was added CuBr2 (713 mg, 3.19 mmol, 1.5 eq), acetonitrile (10 mL) and tert-butyl nitrite (500 mg, 4.68 mmol, 2.2 eq), the mixture was controlled under 60 °C in the nitrogen atmosphere. The reaction was kept at the temperature for 20min. Next, a solution of 2,4-difluoro-6-nitro-phenylamine (371 mg.2.13 mmol,l eq) in acetonitrile (10 mL) was slowly added into the mixture. The reaction was stirred for 0.5 h. TLC (PE: ethyl acetate=5:l) showed the reaction was completed. The mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford compound 6A (300 mg, HPLC: 90%) as a yellow solid, Yield: 59.2 %.

[00186] Step 2'. To a 100 mL three necked flask was added compound 6A (300 mg, 1.26 mmol, 1 eq), CS ₂ CO ₃ (814 mg, 2.5 mmol, 2 eq), 1,4-dioxane (10 mL), piperidine (150 mg, 1.7 mmol, 1.4 eq), Xantphots (37 mg, 0.06 mmol, 0.05 eq) and palladium diacetate (14 mg, 0.06 mmol, 0.05 eq). The reaction was heated to reflux and stirred for 16 h. TLC (PE: ethyl acetate=5:l) showed the reaction was completed. The mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford compound 6B (250 mg, HPLC: 90%) as a yellow solid, Yield: 82.0 %.

[00187] Step 3. To a 100 mL round-bottom flask was added compound 6B (250 mg, 1.03 mmol, 1 eq), methanol (40 mL), and Raney Ni (1.8 g, 21.6 mmol, 1.5 eq) under Hydrogen atmosphere. To this well-stirred solution, the reaction mixture was heated to 50 °C for 1 h, TLC (PE: ethyl acetate=5:l) showed the reaction was completed. The reaction was allowed to cool at room temperature, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure and purified by column chromatography on silica gel to the compound 6C (200 mg, HPLC: 95%) as a colorless oil. Yield: 91.5%.

[00188] Step 4. To a 100 mL round-bottom flask was added compound 6C (86.7 mg, 0.4 mmol, 1.0 eq), compound 4D (130 mg, 0.45 mmol, 1.1 eq) and pyridine (10 mL). The reaction was stirred for 20 h at room temperature (20-30 °C). TLC (PE: ethyl acetate=5:l) showed the reaction was completed. The mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford compound 6 (30 mg, HPLC: 92%) as a brown solid. Yield: 16.1%. Overall Yield: 7.2%. MS-ESL [M+l] ⁺ = 465.9. *HNMR (400 MHz, CDCI ₃ ) δ 7.58 (d, 1H), 7.41 (d, 1H), 7.27 - 7.23 (m, 1H), 6.61 (ddd, 1H), 3.06 (t, 2H), 2.78 (s, 6H), 2.51-2.43 (m, 2H), 1.90 - 1.75 (m, 2H), 1.70-1.60 (m, 2H), 1.41 - 1.30 (m, 2H)

Example 7: Synthesis of N2-[3-chloro-2-(4, 4-difhioro-l-piperidyl) phenyl] -N5, N5-dimethyl thiophene-2, 5-disulfonamide

[00189] Step 1. To a 100 mL round-bottom flask was added 20mL dimethyl sulfoxide, 1-chloro- 2-fluoro-3 -nitrobenzene (1.12 g, 6.38 mmol, 1.0 eq) and 4, 4-difluoropiperidine Hydrochloride (1.11 g, 7.01 mmol, 1.1 eq), K ₂ CO ₃ (2.2 g, 16 mmol, 2.5 eq), the reaction mixture was heated to 140 °C and stirred overnight, TLC (PE: ethyl acetate=10:l, Rf=0.5) showed the reaction was completed. The reaction was allowed to cool to room temperature and100mL water was added to the mixture. The mixture extracted with ethyl acetate (50 mL*3) , the combined organic layer was washed with brine (100 mL*l), dried over Na ₂ SO ₄ , filtered and concentrated under vacuum to afford the compound 7A (0.8 g, HPLC: 95%) as a yellow oil. Yield: 45.4%.

[00190] Step 2'. To a 100 mL round-bottom flask was added 20 mL methanol, compound 7A (0.8 g, 2.89 mmol, 1.0 eq), 0.3 g Raney Ni, and the reaction was heated to 30 °C for 1 h, TLC (PE: ethyl acetate=10:l, Rf=0.2) showed the reaction was completed. The reaction was allowed to cool at room temperature, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure and purified by column chromatography on silica gel to afford the title compound 7B (0.4 g, HPLC: 95%) as a brown-red oil. Yield: 56.4%.

[00191] Step 3. To a 50 mL round-bottom flask was added 4mL pyridine, compound 7B (70 mg, 0.28 mmol, 1.0 eq) and compound 4D (100 mg, 0.34 mmol, 1.2 eq), the reaction was stirred at 30°C overnight. TLC (PE: ethyl acetate=5:l, Rf=0.3) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound (30 mg, HPLC: 94%) as an off-white solid. Yield: 21.4%. MS-ESI: [M-l] ’ = 497.7. ¹ H NMR (400 MHz, CDCI ₃ ) δ 8.55 (s, 1H), 7.60 (dd, 1H), 7.53 (d, 1H), 7.40 (d, 1H), 7.21 (t, 1H), 7.12 (dd, 1H), 3.75-3.68 (m, 2H), 2.79 (s, 6H), 2.53 - 2.38 (m, 2H), 2.20-2.14 (m, 2H), 2.12 - 1.92 (m, 2H).

Example 8: Synthesis of N5-[3-Chloro-2-(l-piperidyl)phenyl]-N2,N2-dimethylthiazole- 2,5- disulfonamide

[00192] Step 1. To a 250 mL three-necked flask was added 120 mL diethyl ether and 2- bromothiazole (5.0 g, 0.030 mol, 1.0 eq) which was degassed with N2 three times. n-BuLi (2.5M) (13.4 mL, 0.033 mol, 1.1 eq) was dropped into the stirred mixture (T< -78 °C). After stirring for 1 h at -78 °C, sulfur dioxide solution in diethyl ether (200 g/L) (48 mL, 0.15 mol, 5.0 eq) was dropped into the stirred mixture (T< -78 °C). The reaction mixture was warmed to room temperature slowly and then concentrated under vacuum to afford the compound 8A (10 g, crude).

[00193] Step 2\ To a 250 mL round-bottom flask was added 125 mL dichloromethane, compound 8A (4.65 g, 0.030 mol, 1.0 eq) and N-chlorosuccinimide (4.5 g, 0.033 mol, 1.1 eq). The mixture was stirred at room temperature for 30 min with N ₂ and then filtered. The filtrate was concentrated under vacuum to afford the compound 8B (7.0 g, crude).

[00194] Step 3: To a 100 mL round-bottom flask was added 20mL tetrahydrofuran, compound 8B (5.59 g, 0.030 mol, 1.0 eq) and dimethylamine hydrochloride (2.98 g, 0.036 mol, 1.2 eq). Triethylamine (15.4 g, 0.15 mol, 5.0 eq) was dropped to the reaction at 10 °C. The reaction was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 5: 1) showed the reaction was completed. The reaction mixture was added100mL water and extracted with ethyl acetate (50 mL*2). The organic layer was dried, concentrated under vacuum and purified by column chromatography on silica gel to afford the title compound 8C (1.2 g, HPLC: 96%) as a yellow oil. Yield (three steps): 20.6%.

[00195] Step 4: To a 100mL three-necked flask was added 40mL tetrahydrofuran and compound 8C (1.0 g, 5.2 mmol, 1.0 eq) which was degassed with N2 three times. n-BuLi (2.5 M) (2.3 mL, 5.73 mol, 1.1 eq) was dropped into the stirred mixture (T< -78 °C). After stirring for Ih at -78 °C, sulfur dioxide solution in diethyl ether (200 g/L) (8.5 mL, 26 mmol, 5.0 eq) was dropped into the stirred mixture (T< -78 °C). The reaction mixture was warmed to room temperature slowly and then concentrated under vacuum to afford the compound 8D (2 g, crude).

[00196] Step 5: 40 mL dichloromethane, compound 8D (1.36 g, 5.2 mmol, 1.0 eq) and N- chlorosuccinimide (1.39 g, 10.4 mmol, 2.0 eq) were added to a 100 mL round-bottom flask. The mixture was stirred at room temperature for 1 h with N2 and then filtered. The filtrate was concentrated under vacuum to afford the compound 8E (2.1 g, crude).

[00197] Step 6: To a 25 mL round-bottom flask was added 1 mL pyridine, compound 5B (100 mg, 0.48 mmol, 1.0 q) and compound 8E (276 mg, 0.95 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 5: 1) showed the reaction was completed. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica gel to afford the mixture of compound 8 (51 mg, HPLC: 91%). Yield (three steps): 16.2%. Overall Yield: 3.3%. MS-ESI: [M+l] ⁺ = 464.8. ¹ H NMR (400 MHz, DMSO): 5 9.89 (s, 1H), 8.55 (s, 1H), 7.34 - 7.29 (m, 1H), 7.19 - 7.15 (m, 2H), 3.26 - 3.03 (m, 2H), 2.86 (s, 6H), 2.65 - 2.53 (m, 2H), 1.64 - 1.38 (m, 6H).

Example 9: Synthesis of N2-[3-chloro-2-(l-piperidyl)phenyl]-N5,N5-dimethylthiazole-2 ,5- disulfonamide

[00198] Step 1. To a 100 mL round-bottom flask was added 5 mL pyridine, compound 8B (8.0 g, 30 mmol, 1.2 eq) and compound 5B (4.4 g, 25 mmol, 1 eq), the reaction was stirred at 50 °C for 3 h. TLC (PE: ethyl acetate=l : 1) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 9A (3.5 g, HPLC: 90%) as a white solid. Yield: 30.0%

[00199] Step 2\ To a 100 mL three necked flask was added 20 mL tetrahydrofuran, compound 9A (1g, 3mmol, l.Oeq), the mixture controlled to -70-80 °C, then n-BuLi (6.2 mL, 5 mmol, 2.2 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the mixture was stirred for 1 h under -70-80°C, then sulfur dioxide solution in diethyl ether (5 mL, 15 mmol, 5 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature and then concentrated under reduced pressure to obtain the crude compound 9B.

[00200] Step 3. To a 100 mL three necked flask was added 40mL di chloromethane, compound 9B, then N-chlorosuccinimide (440 mg, 3.3 mmol, 1.1 eq) was added to the reaction mixture in portion under room temperature, then the reaction mixture was stirred 1 h, TLC (PE: ethyl acetate=L) showed the reaction was completed. The mixture was filter and the filtrate was concentrated under vacuum to yield the crude compound 9C as yellow oil.

[00201] Step 4. To a 50 mL round-bottom flask was added 6 mL dichloromethane, compound 9C (1.5 g, 3 mmol, 1.0 eq), dimethylamine hydrochloride (244.6 mg, 3 mmol, 1 eq) and triethylamine (1.5 g, 15 mmol, 5 eq). The reaction was stirred at 25°C overnight. TLC (PE: ethyl acetate=l :l) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 9 (11.6 mg, HPLC: 90%) as an off-white solid. Yield: 1.0%.

[00202] Overall Yield: 0.3%. MS-ESI: [M+l] ⁺ = 464.6. ¹ HNMR (400 MHz, CDCI ₃ ): δ 9.56 (br, 1H), 8.17 (s, 1H), 7.62 (d, 1H), 7.14-7.08 (m, 2H), 3.49 (t, 2H), 2.81 (s, 6H), 2.64 (m, 2H), 1.95-1.7 (m, 5H), 1.45-1.33 (m, 1H).

Example 10: Synthesis of N2-[3-chloro-2-(4-methyl-l-piperidyl)phenyl]-N5,N5- dimethylthiophene-2,5-disulfonamide

[00203] Step 1. To a 100 mL three necked flask was added 50 mL 1,4-dioxane. To the stirred mixture was added l-Chloro-2-fluoro-3 -nitro-benzene (5.0 g, 28.5 mmol, 1.0 eq), 4-Methyl- piperidine (3.1 g, 31.3 mmol, 1.1 eq) and K ₂ CO ₃ (7.9 g, 57 mmol, 2 eq). The reaction was heated to reflux overnight. TLC(PE) showed the reaction was completed. The reaction was allowed to cool at room temperature. The reaction mixture filtered and concentrated under vacuum, the crude compound was purified by column chromatography on silica gel to afford compound 10A (6 g, HPLC:92%) as a yellow oil. Yield: 82.5 %.

[00204] Step 2 To a 50 mL round-bottom flask was added 20 mL methanol. To the stirred mixture was added compound 10A (2 g, 7.8 mmol, 1.0 eq), Raney Ni (1.0 g). The reaction was degassed with hydrogen three times and stirred at room temperature overnight. TLC (PE: ethyl acetate=5:l) showed the reaction was completed. The reaction was filtered, the filtrate was concentrated under vacuum and the crude compound was purified by column chromatography on silica gel to afford compound 10B (0.8 g, HPLC: 95%) as a yellow oil. Yield: 45.5%.

[00205] Step 3 To a 50 mL round-bottom flask was added 1.5 mL pyridine. To the stirred mixture was added compound 10B (89.9 mg, 0.4 mmol, 1.0 eq), compound 4D (150 mg, 0.52 mmol, 1.3 eq). The reaction was stirred at 20 °C-30 °C overnight. TLC (PE: ethyl acetate=5:l) showed the reaction was completed. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica gel to afford the compound 10 (50 mg, HPLC: 92 %) as a yellow solid. Yield: 26.1%. Overall Yield: 9.8%. MS-ESI: [M+l] ⁺ = 477.9. ¹ H NMR (400 MHz, CDCI ₃ ) δ 9.04 (br, 1H), 7.61 (dd, 1H), 7.54 (d, 1H), 7.39 (d, 1H), 7.18 (t, 1H), 7.10 (dd, 1H), 3.48 (t, 2H), 2.75 (s, 6H), 2.45-2.32 (m, 2H), 1.79-1.70 (m, 2H), 1.60-1.50 (m, 1H), 1.38 - 1.29 (m, 2H), 1.03 (d, 3H).

Example 11: Synthesis of N2-[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] -N5, N5-dimethyl thiophene-2, 5-disulfonamide

[00206] Step 1. To a 500 mL three necked flask was added 250 mL tetrahydrofuran, and lithium aluminium hydride (6.6 g, 150 mmol, 3.0 eq) was added to the reaction mixture in portions under 10- 15 °C, then 3,3-Dimethylglutarimide (7.05 g, 50 mmol, 1.0 eq) in 100 mL tetrahydrofuran was added to the reaction mixture, then the reaction mixture was stirred at room temperature for 0.5 h, then reflux for 3 h. LC-MS showed the reaction was completed, the mixture was cooled to 5 °C and 6.6 mL water, 13mL 15% sodium hydroxide aqueous solution and 6.6 mL water was added to the reaction mixture in portions. Then the mixture was stirred for 0.5 h, filtered and the filtrate was added into 500 mL three necked flask, then 40 mL HC1 EtOH solution was added to the mixture, the mixture was stirred for Ih, the mixture was concentrated under vacuum to afford the compound 11A (4.0 g) as an off-white solid. Yield: 70.6%.

[00207] Step 2\ To a 500 mL round-bottom flask was added 100 mLl, 4-dioxane, l-chloro-2- fluoro-3 -nitrobenzene (4.2 g, 24.2 mmol, 1.0 eq) and compound 11A (4.0 g, 26.6 mmol, 1.1 eq), K ₂ CO ₃ (11 g, 72.7 mmol, 3.0 eq), the reaction mixture was heated to reflux overnight, TLC (PE: ethyl acetate=10:l, Rf=0.7) showed the reaction was completed. The mixture was cooled and concentrated under vacuum and the resulting compound was purified by column chromatography on silica gel to afford the title compound 11B (3.4 g, HPLC: 90%) as an orange-red solid. Yield: 52.3%. [00208] Step 3. To a 50 mL round-bottom flask was added 20 mL methanol, compound 11B (1.4 g, 5.22 mmol, 1.0 eq), 1.0 g Raney Ni, and the reaction was heated to 50 °C for 2.5 h, TLC (PE: ethyl acetate=10:l, Rf=0.6) showed the reaction was completed. The reaction was allowed to cool at room temperature, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure and purified by column chromatography on silica gel to afford the title compound 11C (850 mg, HPLC: 95%) as an off-white oil. Yield: 70.8%.

[00209] Step 4. To a 50 mL round-bottom flask was added 2 mL pyridine, compound 11C (95 mg, 0.40 mmol, 1.0 eq) and compound 4D (150 mg, 0.52 mmol, 1.3 eq), the reaction was stirred at 30°C overnight. TLC (PE: ethyl acetate=5:l, Rf=0.3) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 11 (70 mg, HPLC: 98.5%) as an off-white solid. Yield: 35.5%. MS- ESI: [M-l] ’=492.1, 494.1. ¹ H NMR (400 MHz, CDCI ₃ ) δ 9.02 (s, IH), 7.61 (dd, IH), 7.52 (d, IH), 7.38 (d, IH), 7.18 (t, IH), 7.11 (dd, IH), 3.62 (td, 2H), 2.75 (s, 6H), 2.19-2.10 (m, 2H), 1.54 (td, 2H), 1.48-1.37 (m, 2H), 1.04 (s, 6H).

Example 12: Synthesis of N4,N4-dimethyl-N2-[2-(l-piperidyl)phenyl] thiophene-2, 4- disulfonamide

[00210] Step 1. To a 500 mL round-bottom flask was added 200 mL toluene, 3 -bromo thiophene (10 g, 61.3 mmol, 1.0 eq), benzyl mercaptan (7.62 g, 61.3 mmol, 1.0 eq), N,N-diisopropylethylamine (15.85 g, 0.123 mmol, 2.0 eq), Xantphos (2.13 g, 3.68 mmol, 0.06 eq) and Pd(dppf)C12.CHCls (1.50 g, 1.84 mmol, 0.03 eq). The reaction was heated to 110 °C for 3 h with nitrogen. TLC (PE) showed the reaction was completed. The reaction mixture was cooled, concentrated under vacuum and purified by column chromatography on silica gel to afford the compound 12A (11.6 g) as a colorless oil. Yield: 92.1%.

[00211] Step 2\ To a 500 mL round-bottom flask was added 200 mL glacial acetic acid, 60 mL water and compound 12A (11.6 g, 56.3 mmol, 1.0 eq). N-chlorosuccinimide (30.1 g, 225 mmol, 4.0 eq) was added to the stirred mixture at 10 °C. The reaction was stirred at room temperature for 1 h with nitrogen. TLC (PE) showed the reaction was completed. Then the mixture was concentrated and purified by column chromatography on silica gel to afford the title compound 12B (13.2 g) as a colorless oil.

[00212] Step 3: To a 500 mL round-bottom flask was added 200mL dichloromethane, compound 12B (10.3 g, 56.3 mmol, 1.0 eq), dimethylamine hydrochloride (5.52 g, 67.7 mmol, 1.2 eq). Triethylamine (28.6 g, 0.282 mol, 5.0 eq) was dropped to the stirred mixture at 10 °C. The reaction was stirred at room temperature for 16 h with nitrogen. TLC (PE: ethyl acetate= 10: 1) showed the reaction was completed. The reaction mixture was added 200 mL water and extracted with dichloromethane (100 mL*2). The organic layer was dried, concentrated under vacuum and purified by column chromatography on silica gel to afford the compound 12C (8.1 g, HPLC: 92%) as a white solid. Yield (two steps): 75.7%. [00213] Step 4. To a 250 mL round-bottom flask was added 80 mL glacial acetic acid and compound 12C (8.1 g, 42.4 mmol, 1.0 eq). N-bromosuccinimide (11.3 g, 63.6 mmol, 1.5 eq) was added to the stirred mixture. The reaction was stirred at 100 °C for 2 h with nitrogen. The reaction mixture was cooled, added 500 mL water and extracted with ethyl acetate (200 mL*2). The organic layer was dried, concentrated under vacuum and purified by column chromatography on silica gel to afford the compound 12D (3.6 g, HPLC: 96%) as a solid. Yield: 31.5%.

[00214] Step 5: To a 100 mL round-bottom flask was added 20 mL toluene, compound 12D (1 g, 3.70 mmol, 1.0 eq), benzyl mercaptan (460 mg, 3.70 mmol, 1.0 eq), N,N-diisopropylethylamine (957 mg, 7.41 mmol, 2.0 eq), Xantphos (129 mg, 0.22 mmol, 0.06 eq) and Pd(dppf)C12.CHCh (91 mg, 0.11 mmol, 0.03 eq). The reaction was heated to 110 °C for 3 h with nitrogen. The reaction mixture was cooled, concentrated under vacuum and purified by column chromatography on silica gel to afford the compound 12E (1.0 g, HPLC: 89%) as a solid. Yield: 86.2%.

[00215] Step 6: To a 50 mL round-bottom flask was added lOmL glacial acetic acid, 3 mL water and compound 12E (500 mg, 1.6 mmol, 1.0 eq). N-chlorosuccinimide (853 mg, 6.4 mmol, 4.0 eq) was added to the stirred mixture at 10 °C. The reaction was stirred at room temperature for 1 h with nitrogen. TLC (PE: ethyl acetate= 5: 1) showed the reaction was completed. Then the mixture was concentrated and purified by column chromatography on silica gel eluted with PE: ethyl acetate= 5: 1 to afford the compound 12F (438 mg, HPLC: 77%) as a white solid. Yield: 94.8%.

[00216] Step 7: To a 25 mL round-bottom flask was added ImL pyridine, compound 4F (100 mg, 0.68 mmol, 1.0 eq) and compound 12F (197 mg, 0.68 mmol, 1.2 eq). The reaction mixture was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 5: 1) showed the reaction was completed. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica gel to afford the compound 12 (98 mg, HPLC: 98%). Yield: 40.3%.

Overall Yield: 7.2%. MS-ESI: [M+l] ⁺ = 430.5. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.48 - 8.28 (m, 1H), 7.99 (s, 1H) 7.72 - 7.61 (m, 2H), 7.24 - 7.09 (m, 3H), 2.68 (s, 6H), 2.63 - 2.42 (m, 4H), 1.82 - 1.50 (m, 6H).

Example 13: Synthesis of N2,N2-dimethyl-N4-[2-(l-piperidyl)phenyl] thiophene- 2,4- disulfonamide

[00217] Step 1. To a 500 mL round-bottom flask was added 200 mL dimethyl sulfoxide, 2,3,5- Tribromothiophene (25.0 g, 77.9 mmol, 1.0 eq), the mixture controlled to 15-20 °C. Then NaBFL (6 g, 158.6 mmol, 2 eq) was added to the reaction mixture in the nitrogen atmosphere. The reaction was stirred at 20-25 °C for 16 h. TLC (PE) showed the reaction was completed. The reaction mixture was quenched in water 400 mL at 10-15 °C and extracted with tert-butyl methyl ether (200 mL*3). The combined organic layer was washed with water (200 mL). The organic layer was dried with Na ₂ SO ₄ and concentrated under reduced pressure to afford compound 13A (11 g, GC: 92%) as a white oil. Yield: 58.3%

[00218] Step 2\ To a 100 mL three necked flask was added 40 mL tetrahydrofuran and compound 13A (1.5 g, 5 mmol, 1.0 eq), the mixture was cooled to -30~ -40 °C. Then z-PrMgBr (2.75 mL, 5.5 mmol, 1.1 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the mixture was stirred for 1 h under -30~ -40 °C, then sulfur dioxide solution in diethyl ether (8 mL, 25 mmol, 5 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature and then concentrated under reduced pressure to obtain the crude compound 13B.

[00219] Step 3: To a 100 mL three necked flask was added 40 mL di chloromethane, compound 13B, then N-chlorosuccinimide (0.79 g, 5.5 mmol, 1.1 eq) was added to the reaction mixture in portion under room temperature, then the reaction mixture was stirred 1 h, TLC (PE) showed the reaction was completed. The mixture was filter and the filtrate was concentrated under vacuum to yield the crude compound 13C (2.0 g) as a yellow oil.

[00220] Step 4. To a 50 mL round-bottom flask was added 6 mL dichloromethane, compound 13C (2.0 g, 0.55 mmol, 1 eq), Dimethylamine hydrochloride (447 mg, 0.55 mmol, 1.1 eq) and triethylamine (757 mg, 0.75 mmol, 1.5 eq). The reaction was stirred at 25 °C overnight. TLC (PE:ethyl acetate=l:l) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 13D (0.7 g, HPLC: 90%) as an off-white solid. Yield: 50.0%.

[00221] Step 5: To a100mL three necked flask was added 20 mL tetrahydrofuran, compound 13D (0.7 g, 2.5 mmol, 1.0 eq), the mixture controlled to -70~ -80 °C, then n-BuLi (2 mL, 5mmol, 2 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the mixture was stirred for 1 h under -70~ -80 °C, then sulfur dioxide solution in diethyl ether (10 mL, 5Vol) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature and then concentrated under reduced pressure to obtain the crude compound 13E.

[00222] Step 6: To a 100 mL three necked flask was added 40 mL di chloromethane, compound 13E, then N-chlorosuccinimide (367 mg, 2.75 mmol, 1.1 eq) was added to the reaction mixture in portion under room temperature, then the reaction mixture was stirred Ih, TLC (PE: ethyl acetate=l:l) showed the reaction was completed. The mixture was filter and the filtrate was concentrated under vacuum to yield the crude compound 13F (0.9 g) as a yellow oil.

[00223] Step 7: To a 100 mL round-bottom flask was added 5 mL pyridine, compound 13F (0.9 g, 2.5 mmol, 1.2 eq) and 2-Piperidin-l-yl-phenylamine (366 mg, 2.08 mmol, 1 eq), the reaction was stirred at 50 °C for 3 h. TLC (PE: ethyl acetate=l : 1) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the compound 13 (19.6 mg, HPLC: 86%) as ayellow oil. Yield: 1.8%. Overall Yield: 0.52%. MS-ESE [M+l] ⁺ = 429.8. *HNMR (400 MHz, CDCI ₃ ): 8 8.38 (s, IH), 7.65 (d, IH), 7.57- 7.47 (m, IH), 7.38 (s, IH), 7.25-7.10 (m, 3H), 2.78 (s, 6H), 2.65-2.40 (m, 4H), 1.75-1.55 (m, 6H).

Example 15: Synthesis of N4-[2-(4,4-dimethyl-l-piperidyl)phenyl]-N2,N2- dimethylthiophene- 2,4-disulfonamide

[00224] Step 1. To a 500 mL three necked flask was added 250 mL tetrahydrofuran, lithium aluminum hydride(3.42 g, 90 mmol, 3.0 eq) was added to the reaction mixture in portion under 10-15 °C, then 4, 4-dimethylazaperhydroine-2, 6-dione (4.33 g, 30 mmol, 1.0 eq) in 100 mL tetrahydrofuran was added to the reaction mixture, then the reaction mixture was stirred at room temperature for 0.5 h, then reflux for 3 h. LC-MS showed the reaction was completed, the mixture was cooled to 5 °C and 3.5 mL water, 7 mL 15% sodium hydroxide aqueous solution and 3.5 mL water was added to the reaction mixture in portions. Then the mixture was stirred for 0.5 h, vacuum filtration, the filtrate was added 500 mL three necked flask, then 40 mL HCl/EtOH was added to the mixture, the mixture was stirred for 1 h, the mixture was concentrated under vacuum to afford the compound 15A (3.5 g) as an off-white solid. Yield: 78.8%.

[00225] Step 2'. To a 500 mL round-bottom flask was added 100 mLl,4-dioxane, l-Fluoro-2- nitrobenzene (2.6 g, 18.8 mmol, 0.8 eq) and 15A (3.5 g, 23.6 mmol, 1.1 eq), K ₂ CO ₃ (9.7 g, 70.8 mmol, 3.0 eq), the reaction mixture was heated to reflux overnight, TLC (PE: ethyl acetate=10:l) showed the reaction was completed. The mixture was cooled and concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 15B (2.3 g, HPLC: 95%) as an orange-red solid. Yield: 43.4%.

[00226] Step 3. To a 50 mL round-bottom flask was added 20 mL methanol, compound 15B (2.3 g, 10 mmol, 1.0 eq), 1.0 g Raney Ni, and the reaction was heated to 50 °C for 2.5 h, TLC (PE: ethyl acetate=10:l) showed the reaction was completed, the reaction was allowed to cool at room temperature, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure and purified by column chromatography on silica gel to afford the title compound 15C (1.8 g, HPLC: 95%) as a yellow solid. Yield: 88%.

[00227] Step 4. To a 50 mL round-bottom flask was added 2 mL pyridine, compound 15C (108 mg, 0.53 mmol, 1.0 eq) and compound 13F (200 mg, 0.7 mmol, 1.3 eq), the reaction was stirred at 30 °C overnight. TLC (PE: ethyl acetate=5:l) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the compound 15 (104 mg, HPLC: 97%) as an off-white solid. Yield: 42.9 %. All Yield: 3.1%. MS-ESI: [M+l] ⁺ =458.5. ¹ H NMR (400 MHz, CDCI ₃ ): 8.22 (s, 1H), 8.11 (s, 1H), 7.61 (d, 2H), 7.25- 7.05(m, 3H), 2.72 (s, 6H), 2.65-2.4 (m, 4H), 1.65-1.4 (m, 4H), 1.04 (s, 6H).

Example 16: Synthesis of N5-[2-(4,4-dimethyl-l-piperidyl)phenyl]-N2,N2-dimethylthiazo le- 2,5- disulfonamide

[00228] Step 1. To a 25 mL round-bottom flask was added 1 mL pyridine, compound 15C (200 mg, 0.98 mmol, 1.0 q) and compound 8E (712 mg, 2.45 mmol, 2.5 eq). The reaction mixture was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 5: 1) showed the reaction was completed. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica gel to afford the mixture of compound 16 (55 mg, HPLC: 95.9%). Yield: 12.2%. MS-ESI: [M+l] ⁺ = 459.5. ¹ H NMR (400 MHz, DMSO): δ 9.55 (br, 1H), 8.46 (s, 1H), 7.36 - 7.23 (m, 2H), 7.19 (t, 1H), 7.11 (t, 1H), 2.84 (s, 6H), 2.71 - 2.61 (m, 4H), 1.45 - 1.31 (m, 4H), 0.94 (s, 6H).

Example 17: Synthesis of N2-[2-(4, 4-dimethyl-l-piperidyl) phenyl]-N5, N5-dimethylthiophene- 2, 5-disulfonamide [00229] Step 1. To a 50 mL round-bottom flask was added 4 mL pyridine, compound 15C (89 mg, 0.43 mmol, 1.0 eq) and compound 4D (150 mg, 0.52 mmol, 1.2 eq), the reaction was stirred at 25 °C overnight. TLC (PE: ethyl acetate=3:l, Rf=0.5) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 17 (47 mg, HPLC: 98.7%) as an off-white solid. Yield: 23.9%. MS-ESI: [M+l] ⁺ =458.5. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.36 (s, 1H), 7.65 (d, 1H), 7.53 (s, 1H), 7.37 (d, 1H), 7.24-7.14 (m, 3H), 2.76 (s, 6H), 2.54 (br, 4H), 1.50 (br, 4H), 1.03 (s, 6H).

Example 18: Synthesis of N2-[2-(7-azaspiro [3.5] nonan-7-yl) phenyl] -N5, N5- dimethylthiophene-2, 5-disulfonamide

[00230] Step 1. To a 100 mL round-bottom flask was added 15 mL 1,4-dioxane, l-fluoro-2- nitrobenzene (831 mg, 5.89 mmol, 1.0 eq) and 7-azaspiro[3.5]nonane hydrochloride (1 g, 6.18 mmol, 1.05 eq) and K ₂ CO ₃ (2.1 g, 15.45 mmol, 2.5 eq), the reaction mixture was heated to reflux overnight, TLC (PE: ethyl acetate=10:l, Rf=0.7) showed the reaction was completed. The mixture was cooled and concentrated under vacuum to yield the crude compound 18A (1.8 g, HPLC: 88%) as an orange oil.

[00231] Step 2'. To a 100 mL round-bottom flask was added 40 mL methanol, compound 18A (1.8 g), 0.5 g Raney Ni, and the reaction was stirred at room temperature overnight, TLC (PE: ethyl acetate=10:l, Rf=0.5) showed the reaction was completed, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure and purified by column chromatography on silica gel to afford the title compound 18B (0.7 g, HPLC: 95%) as an oil. Yield: 55.1% (two steps). [00232] Step 3: To a 50 mL round-bottom flask was added 5 mL pyridine, compound 18B (103 mg, 0.48 mmol, 1.0 eq) and compound 4D (179 mg, 0.62 mmol, 1.3 eq), the reaction was stirred at 45°C for 4 h. TLC (PE: ethyl acetate=3:l, Rf=0.4) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title product (105 mg, HPLC: 99.3%) as a light-yellow solid. Yield: 46.7%. MS-ESI: [M+l] ⁺ =470.5. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.37 (s, 1H), 7.64 (d, 1H), 7.51 (d, 1H), 7.37 (d, 1H), 7.18-7.10 (m, 3H), 2.76 (s, 6H), 2.50-2.42 (m, 4H), 1.99 - 1.88 (m, 2H), 1.84-1.81 (m, 4H), 1.77-1.65 (m, 4H).

Example 19: Synthesis of N4-[2-(7-azaspiro[3.5]nonan-7-yl)phenyl]-N2,N2- dimethylthiophene- 2,4-disulfonamide

[00233] Step 1. To a 50 mL round-bottom flask was added 2 mL pyridine, compound 18B (116 mg, 0.53 mmol, 1.0 eq) and compound 13F (200 mg, 0.7 mmol, 1.3 eq), the reaction was stirred at 15-25°C overnight. TLC (PE: ethyl acetate=5:l) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the compound 19 (110 mg, HPLC: 97%) as a yellow solid. Yield: 44.3 %. MS-ESI: [M+l] ⁺ =470.5. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.23 (s, 1H), 8.10 (d, 1H), 7.60 (d, 2H), 7.25-7.05 (m, 3H), 2.73 (s, 6H), 2.50-2.40 (m, 4H), 1.99 - 1.64 (m, 10H).

Example 20: Synthesis of N2-[2-(8-azaspiro[4.5]decan-8-yl)phenyl]-N5,N5- dimethylthiophene- 2,5-disulfonamide [00234] Step 1. To a 250 mL three necked flask was added 35 mL tetrahydrofuran, lithium aluminum hydride (3.4 g, 89.7 mmol, 3.0 eq) was added to the reaction mixture in portion under 10- 15 °C, then 8-Aza-spiro[4.5]decane-7, 9-dione (5 g, 29.9 mmol, 1.0 eq) in 50 mL tetrahydrofuran was added to the reaction mixture, then the reaction mixture was stirred at room temperature for 0.5 h, then the mixture was cooled to 5 °C and 3.4 mL water, 6.8 mL 15% sodium hydroxide aqueous solution and 3.4 mL water was added to the reaction mixture in portions. Then the mixture was stirred for 0.5 h, filtered, and the filtrate was added to100mL three necked flask, then 40 mL HCl/EtOH was added to the mixture, the mixture was stirred for Ih, concentrated under vacuum to afford the compound 20A (5 g) as an off-white solid. Yield: 100%.

[00235] Step 2\ To a 250 mL round-bottom flask was added 100 mLl,4-dioxane, compound 20A (5 g, 29.9 mmol, 1.1 eq) and 1 -fluoro-2-nitrobenzene (3.85 g, 27.2 mmol, 1 eq), K ₂ CO ₃ (12.4 g, 89.7 mmol, 3.0 eq), the reaction mixture was heated to reflux overnight, TLC (PE: ethyl acetate=10:l) showed the reaction was completed. The mixture was cooled and concentrated under vacuum and the resulting product was purified by column chromatography on silica gel to afford the title compound 20B (4.7 g, HPLC: 92%) as an orange-red solid. Yield: 66.6%.

[00236] Step 3. To a 100 mL round-bottom flask was added 27 mL methanol, compound 20B (2.7 g, 10.4 mmol, 1.0 eq), 0.9 g Raney Ni, and the reaction was heated to 50 °C overnight, TLC (PE: ethyl acetate=10:l) showed the reaction was completed, the reaction was allowed to cool at room temperature, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure and purified by column chromatography on silica gel to afford the title compound 20C (1 g, HPLC: 95%) as an off-white solid. Yield: 42%.

[00237] Step 4. To a 50 mL round-bottom flask was added 4 mL pyridine, compound 20C (99.6 mg, 0.43 mmol, 1.0 eq) and compound 4D (150 mg, 0.52 mmol, 1.2 eq), the reaction was stirred at 30 °C overnight. TLC (PE: ethyl acetate=3:l) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the compound 20 (35 mg, HPLC: 98.5%) as an off-white solid. Yield: 17%. All Yield: 4.7%. MS: [M+l]: 484.5. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.40 (s, IH), 7.64 (d, IH), 7.60-7.47 (m, IH), 7.40-7.35 (m, IH), 7.26-7.05 (m, 3H), 2.76 (s, 6H), 2.65-2.45 (m, 4H), 1.75-1.47 (m, 10H), 135-1.25 (m, 2H).

Example 21: Synthesis of N2-[2-(4, 4-difhioro-l-piperidyl) phenyl] -N5, N5-dimethylthiophene- 2, 5-disulfonamide

[00238] Step 1. To a 100 mL round-bottom flask was added 20 mL 1, 4-dioxane, l-fluoro-2- nitrobenzene (2.26 g, 16.0 mmol, 1.05 eq) and 4,4-difluoropiperidine hydrochloride (2.4 g, 15.2 mmol, 1.0 eq), K ₂ CO ₃ (5.2 g, 38 mmol, 2.5 eq), the reaction mixture was heated to reflux overnight, TLC (PE: ethyl acetate=10:l, Rf=0.4) showed the reaction was completed. The reaction was allowed to cool at room temperature and concentrated under vacuum and the resulting product was purified by column chromatography on silica gel to afford the title compound 21A (3.0 g, HPLC: 92%) as an orange-red solid. Yield: 81.5%.

[00239] Step 2\ To a 250 mL round-bottom flask was added 100 mL methanol, compound 21A (3.0 g, 12.4 mmol, 1.0 eq), 0.8 g Raney Ni, and the reaction was heated to 40 °C for 4 h, TLC (PE: ethyl acetate=10:l, Rf=0.3) showed the reaction was completed, the reaction was allowed to cool at room temperature, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure and purified by column chromatography on silica gel to afford the title compound 21B (1.2 g, HPLC: 95%) as an off-white solid. Yield: 46.1%.

[00240] Step 3: To a 50 mL round-bottom flask was added 4 mL pyridine, compound 21B (150 mg, 0.71 mmol, 1.0 eq) and compound 4D (247 mg, 0.85 mmol, 1.2 eq), the reaction was stirred at 45°C for 2 h. TLC (PE: ethyl acetate=5:l, Rf=0.4) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 21 (120 mg, HPLC: 97%) as an off-white solid. Yield: 36.4%. MS- ESI: [M-l]- =464.4. *HNMR (400 MHz, CDCI ₃ ): δ 8.31 (s, 1H), 7.64 (d, 1H), 7.56 (d, 1H), 7.39 (d, 1H), 7.32 - 7.20 (m, 2H), 7.20 - 7.13 (m, 1H), 2.82-2.78 (m, 10H), 2.26 - 2.10 (m, 4H).

Example 22: Synthesis of N5-[2-(4,4-difluoro-l-piperidyl)phenyl]-N2,N2-dimethylthiazo le- 2,5- disulfonamide

[00241] Step 1. To a 25 mL round-bottom flask was added 0.5 mL pyridine, compound 21B (122 mg, 0.57 mmol, 1.0 q) and compound 8E (200 mg, 0.69 mmol, 1.2 eq). The reaction mixture was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 5: 1) showed the reaction was completed. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica gel to afford the mixture of compound 22 (34 mg, HPLC: 98%). Yield: 12.8%. MS-ESI: [M+l] ⁺ = 467.6. ¹ H NMR (400 MHz, DMSO): 5 10.08 (s, 1H), 8.53 (s, 1H), 7.34 (d, J = 7.8 Hz, 1H), 7.29 - 7.20 (m, 2H), 7.18 - 7.12 (m, 1H), 2.86 (s, 6H), 2.77 - 2.67 (m, 4H), 2.12 - 1.96 (m, 4H).

Example 23: Synthesis of N2-[2-(4, 4-dimethyl-l-piperidyl) phenyl] -N5, N5-dimethylthiazole-2, 5-disulfonamide

[00242] Step 1. To a 250 mL round-bottom flask was added 20 mL pyridine, compound 15C (2.6 g, 12.7 mmol, 1.0 eq) and compound 8B (7.2 g, 1.3 eq, crude product, HPLC: 40%), the reaction was stirred at 30°C overnight. TLC (PE: ethyl acetate=3: l, Rf=0.4) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 23A (570 mg, HPLC: 95%) as an orange-red solid. Yield: 12.8%.

[00243] Step 2\ To a 100 mL three necked flask was added 15 mL tetrahydrofuran, compound 23A (530 mg, 1.51 mmol, 1.0 eq), the mixture was controlled to -70— 80°C, then n-BuLi (1.5 mL, 2.6 mmol, 2.5 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the mixture was stirred for Ih under -70— 80°C, then sulfur dioxide solution in diethyl ether (10 mL, 10 eq, 2.5 M) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature for 1 h and then concentrated under reduced pressure to obtain the crude product.

[00244] To a 100 mL three necked flask was added 30 mL di chloromethane, the crude product, then N-chlorosuccinimide (725 mg, 3.02 mmol, 2.0 eq) was added to the reaction mixture in portion under room temperature, then the reaction mixture was stirred for 1 h, TLC (PE: ethyl acetate=3:l, Rf=0.3) showed the reaction was completed. The mixture was fdter and the filtrate (23B) was used to next step.

[00245] Step 3 To a 50 mL round-bottom flask was added the filtrate of last step (23B), dimethylamine hydrochloride (246 mg, 3.02 mmol, 2.0 eq) and triethylamine (336 mg, 3.32 mmol, 2.2 eq), the reaction was stirred at 5-10°C for 1 h. TLC (PE: ethyl acetate=3:l, Rf=0.2) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 23 (40 mg, HPLC: 94%) as an off- white solid. Yield: 6.0% (two steps). MS-ESE [M+l] ⁺ =459.6. ¹ H NMR (400 MHz, DMSO): 5 10.01 (s, IH), 8.49 (s, IH), 7.46 - 7.36 (m, IH), 7.30 (d, IH), 7.21 - 7.03 (m, 2H), 2.94-2.84 (m, 4H), 2.74 (s, 6H), 1.51 - 1.36 (m, 4H), 0.97 (s, 6H).

Example 24: Synthesis of N2-[2-(4,4-difluoro-l-piperidyl)phenyl]-N4,N4-dimethylthioph ene- 2,4-disulfonamide

[00246] Step 1. To a 25 mL round-bottom flask was added ImL pyridine, compound 21B (100 mg, 0.47 mmol, 1.0 q) and compound 12F (164 mg, 0.57 mmol, 1.2 eq). The reaction mixture was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 10: 1) showed the reaction was completed. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica gel to afford the compound C (51 mg, HPLC: 98%). Yield: 23.3%. MS- ESI: [M+l] ⁺ = 466.5. ¹ H NMR (400 MHz, DMSO): 5 9.68 (s, IH), 8.54 - 8.48 (m, IH), 7.60 -7.55 (m, IH), 7.39 (dd, IH), 7.29 - 7.22 (m, IH), 7.21 - 7.09 (m, 2H), 2.71 - 2.63 (m, 4H), 2.57 (s, 6H), 2.17 - 2.02 (m, 4H).

Example 25: Synthesis of N5-[2-(4, 4-dimethyl-l-piperidyl) phenyl] -N2, N2, 4- trimethylthiazole-2, 5-disulfonamide

[00247] Step 1. To a 250 mL three necked flask was added 50 mL tetrahydrofuran, compound 4- methylthiazole (5 g, 50.4 mmol, 1.0 eq), the mixture was controlled to -70 to -80°C, then n-BuLi (25 mL, 60.5 mmol, 1.2 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the mixture was stirred for Ih under -70 to -80°C, then sulfur dioxide solution in diethyl ether (100 mL, 5.0 eq, 2.5 M) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature for 1 h and then the reaction mixture was concentrated under reduced pressure to obtain the crude product.

[00248] To a 250 mL three necked flask was added 100 mL dichloromethane, the crude product, then N-chlorosuccinimide (24.2 g, 100.8 mmol, 2.0 eq) was added to the reaction mixture in portion under 0-5°C, then the reaction mixture was stirred for 1 h, TLC (PE: ethyl acetate=10:l, Rf=0.15) showed the reaction was completed. The mixture was filter and the filtrate (25A) was used to next step.

[00249] Step 2 To a 250 mL three necked flask was added the filtrate of last step (25A), dimethylamine hydrochloride (5.0 g, 60.5 mmol, 1.2 eq) and triethylamine (7.6 g, 75.6 mmol, 1.5 eq), the reaction was stirred at 5-10°C for 0.5 h. TLC (PE: ethyl acetate=10:l, Rf=0.05) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 25B (4.7 g, HPLC: 96%) as a light yellow solid. Yield: 45.2% (two steps).

[00250] Step 3 To a 100 mL three necked flask was added 24 mL tetrahydrofuran, compound 25B (2.06 g, 10 mmol, 1.0 eq), the mixture was controlled to -70 to -80°C, then n-BuLi (5 mL, 12.0 mmol, 1.2 eq) was added drop wise to the reaction mixture in the nitrogen atmosphere, the mixture was stirred for Ih under -70 to -80°C, then sulfur dioxide solution in diethyl ether (20 mL, 5.0 eq, 2.5 M) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature for 1 h and then the reaction mixture was concentrated under reduced pressure to obtain the crude product.

[00251] To a 100 mL three necked flask was added 50 mL di chloromethane, the crude compound, then N-chlorosuccinimide (4.8 g, 20.0 mmol, 2.0 eq) was added to the reaction mixture in portion under 0-5°C, then the reaction mixture was stirred for 1 h, TLC (PE: ethyl acetate=3:l, Rf=0.4) showed the reaction was completed. The mixture was filter and the filtrate was concentrated under vacuum and the resulting product was purified by column chromatography on silica gel to afford the title compound 25C (1.5g, HPLC: 98%) as a light-yellow solid. Yield: 49.2% (two steps).

[00252] Step 4. To a 50 mL round-bottom flask was added lOmL pyridine, compound 15C (204 mg, 1.0 mmol, 1.0 eq) and compound 25C (396 mg, 1.3 mmol, 1.3 eq), the reaction was stirred at 25°C overnight. TLC (PE: ethyl acetate=3:l, Rf=0.4) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 25 (170 mg, HPLC: 98.5%) as a light-yellow solid. Yield: 36.0%. MS-ESE [M+l] ⁺ =473.7. ¹ H NMR (400 MHz, DMSO): 5 9.85 (s, IH), 7.30 (d, IH), 7.28-7.22 (m, 2H), 7.14 -7.08 (m, IH), 2.82 (s, 6H), 2.62-2.58 (m, 4H), 2.42 (s, 3H), 1.36 - 1.24 (m, 4H), 0.93 (s, 6H).

Example 26: Synthesis of N2,N2,3,4-tetramethyl-N5-[2-(l-piperidyl)phenyl]thiophene-2, 5- disulfonamide

[00253] Step 1. To a 100 mL three-necked flask was added 20 mL diethyl ether and Mg (1.64 g, 0.068 mol, 3.3 eq) which was degassed with N2 three times. CH3I (8.80 g, 0.062 mol, 3.0e q) in 10 mL Diethyl ether was dropped into the stirred mixture slowly at 35 °C. Then the mixture was stirred for 0.5 h at 35 °C. To a 100 mL three-necked flask was added lOmL diethyl ether, 1,3- bis(diphenylphosphino)propane nickel (II) chloride (123 mg, 0.23 mmol, 0.011 eq) and 3,4- dibromothiophene (5.0 g, 0.021 mol, 1.0 eq) which was degassed with N2 three times. MeMgl (30 mL, 0.062 mol, 3.0 eq) was dropped into the stirred mixture. The reaction was heated to reflux for 16 h. The reaction mixture was cooled to 5 °C, added 25 mL IN HC1 and extracted with Diethyl ether (10 mL*2). The organic layer was dried, concentrated under vacuum and purified by distillation at 54 °C at 40 mmHg to afford compound 26A (1.2 g, HPLC: 98%) as a colorless oil. Yield: 52.2%. [00254] Step 2 To a 100 mL three-necked flask was added 10 mL tetrahydrofuran and compound 26A (500 mg, 4.46 mmol, 1.0 eq) which was degassed with N2 three times. n-BuLi (2.5 M) (2.0 mL, 4.91 mmol, 1.1 eq) was dropped into the stirred mixture (T< -78 °C). After stirring for 1 h at -78 °C, sulfur dioxide solution in diethyl ether (220 g/L) (10 mL) was dropped into the stirred mixture (T< -78 °C). The reaction mixture was warmed to room temperature slowly and then concentrated under vacuum to afford the compound 26B (1.8 g, crude).

[00255] Step 3 To a 50 mL round-bottom flask was added 20 mL dichloromethane, compound 26B (812 mg, 4.46 mmol, 1.0 eq) and N-chlorosuccinimide (894 mg, 6.70 mmol, 1.5 eq). The mixture was stirred at room temperature for 30 min with N2 and then filtered. The filtrate was concentrated under vacuum to afford the compound 26C (1.5 g, crude).

[00256] Step 4 To a 50 mL round-bottom flask was added 30 mL dichloromethane, compound 26C (940 mg, 4.46 mmol, 1.0 eq) and dimethylamine hydrochloride (437 mg, 5.36 mmol, 1.2 eq). Triethylamine (2.26 g, 22.3 mmol, 5.0 eq) was dropped to the reaction at 10 °C. The reaction was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 10: 1) showed the reaction was completed. The reaction mixture was added lOmL water and extracted with di chloro methane (10 mL*2). The organic layer was dried, concentrated under vacuum and purified by column chromatography on silica gel eluted with PE: ethyl acetate= 10: 1 to afford the title compound 26D (440 mg, HPLC: 95%) as a white solid. Yield (three steps): 45%.

[00257] Step 5 To a 100 mL three-necked flask was added 10 mL tetrahydrofuran and compound 26D (440 mg, 2.0 mmol, 1.0 eq) which was degassed with N2 three times. n-BuLi (2.5 M) (0.88 mL, 2.2 mmol, 1.1 eq) was dropped into the stirred mixture (T<-78 °C). After stirring for 1 h at -78 °C, sulfur dioxide solution in diethyl ether (220 g/L) (10 mL) was dropped into the stirred mixture (T<-78 °C). The reaction mixture was warmed to room temperature slowly and then concentrated under vacuum to afford the compound 26E (1.8 g, crude). [00258] Step 6 To a 50 mL round-bottom flask was added 20 mL dichloromethane, compound 26E (581 mg, 2.0 mmol, 1.0 eq) and N-chlorosuccinimide (402 mg, 3.0 mmol, 1.5 eq). The mixture was stirred at room temperature for 30 min with N2 and then filtered. The filtrate was concentrated under vacuum and purified by column chromatography on silica gel eluted with PE: ethyl acetate= 5: 1 to afford the compound 26F (170 mg, HPLC: 92%). Yield (two steps): 26.7%.

[00259] Step 7: To a 25 mL round-bottom flask was added 1 mL pyridine, compound 4F (74 mg, 0.42 mmol, 1.0 q) and compound 26F (160 mg, 0.50 mmol, 1.2 eq). The reaction mixture was stirred at room temperature for 16 h. TLC (PE: ethyl acetate= 3: 1) showed the reaction was completed. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica gel eluted with PE: ethyl acetate= 3: 1 to afford the mixture of compound 26 (80 mg, HPLC: 99%). Yield: 41.7%. Overall Yield: 2.6%. MS-ESE [M+l] ⁺ = 458.1. ¹ H NMR (400 MHz, DMSO): 5 9.40 (s, 1H), 7.30 (d, 1H), 7.20 - 7.15(m, 2H), 7.13 - 7.06 (m, 1H), 2.68 (s, 6H), 2.59 - 2.53 (m, 4H), 2.31 (s, 3H), 2.22 (s, 3H), 1.61 - 1.49 (m, 4H), 1.49 - 1.40 (m, 2H).

Example 27: Synthesis of N2-[2-(4, 4-difhioro-l-piperidyl) phenyl] -N4, N4, 3, 5- tetramethylthiophene-2, 4-disulfonamide

[00260] Step 1. To a 100 mL three necked flask was added 40 mL tetrahydrofuran and diisopropylamino (3.8 g, 37.6 mmol, 1.3 eq), n-BuLi (13.7 mL, 34.2 mmol, 1.2 eq) was added to the reaction mixture drop wise under -78 °C, the reaction mixture was stirred for 1 h, then Mel (4.3 g, 37.6 mmol, 1.3 eq) was added to the reaction mixture and stirred at -78 °C for 1 h, then 3-bromo-4- methylthiophene (5 g, 28.2 mmol, 1.0 eq) in 30 mL tetrahydrofuran was added to the reaction mixture and stirred at -78 °C for 1 h. The mixture was stirred at room temperature overnight. HPLC showed the reaction was completed, the mixture was cooled to 5 °C and 30 mL water was added to the reaction mixture in portions, then the mixture was stirred for lOmin, the aqueous phase extracted with di chloromethane (40 mL*3). The organic layer was washed with brine (50 mL*l) and dried over Na ₂ SO ₄ , filtered and concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 27A (3.9 g, HPLC: 93%) as an off-white oil, Yield: 72.9%.

[00261] Step 2 To a 250 mL three necked flask was added 80 mL tetrahydrofuran, compound 27A (2.6 g, 13.7 mmol, 1.0 eq), the mixture was controlled to -70 to -80°C, then n-BuLi (6.7 mL, 16.4 mmol, 1.2 eq) was added dropwise to the reaction mixture in the nitrogen atmosphere, the mixture was stirred for Ih under -70 to -80°C, then sulfur dioxide solution in diethyl ether (30 mL, 5.0 eq, 2.5 M) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature for 1 h and then the reaction mixture was concentrated under reduced pressure to obtain the crude product.

[00262] To a 250 mL three necked flask was added 50 mL dichloromethane, the crude product, then N-chlorosuccinimide (3.7 g, 27.4 mmol, 2.0 eq) was added to the reaction mixture in portion under 0-5°C, then the reaction mixture was stirred for 1 h, TLC (PE: ethyl acetate=5:l, Rf=0.5) showed the reaction was completed. The mixture was filter and the filtrate (27B) was used to next step.

[00263] Step 3 To a 250 mL three necked flask was added the filtrate of last step (27B), dimethylamine hydrochloride (1.34 g, 16.4 mmol, 1.2 eq) and triethylamine (2.1 g, 20.6 mmol, 1.5 eq), the reaction was stirred at 5-10°C for 0.5 h. TLC (PE: ethyl acetate=5:l, Rf=0.3) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 27C (1.1 g, HPLC: 96%) as a white solid. Yield: 36.7% (two steps).

[00264] Step 4. To a 100 mL three necked flask was added 6 mL tetrahydrofuran and diisopropylamino (300 mg, 2.96 mmol, 1.3 eq), then n-BuLi (1.1 mL, 2.74 mmol, 1.2 eq) was added to the reaction mixture under -78 °C in the nitrogen atmosphere, the reaction mixture was stirred for 1 h, then compound 27C (500 mg, 2.28 mmol, 1.0 eq) in 10 mL tetrahydrofuran was added to the reaction mixture and stirred at -78 °C for 1 h. Then sulfur dioxide solution in diethyl ether (5 mL, 5.0 eq, 2.5 M) was added dropwise to the reaction mixture in the nitrogen atmosphere, the reaction was slowly warmed at room temperature for 1 h and then the reaction mixture was concentrated under reduced pressure to obtain the crude product.

[00265] To a 100 mL three necked flask was added 50 mL di chloromethane, the crude product, then N-chlorosuccinimide (608 mg, 4.56 mmol, 2.0 eq) was added to the reaction mixture in portion under 0-5°C, then the reaction mixture was stirred for 1 h, TLC (PE: ethyl acetate=5:l, Rf=0.2) showed the reaction was completed. The mixture was filter and the filtrate was concentrated under vacuum and the resulting product was purified by column chromatography on silica gel to afford the title compound 27D (100 mg, HPLC: 95%) as an off-white solid. Yield: 13.8% (two steps).

[00266] Step 5 To a 50 mL round-bottom flask was added 2 mL pyridine, compound 21B (51 mg, 0.24 mmol, 1.0 eq) and compound 27D (90 mg, 0.28 mmol, 1.2 eq), the reaction was stirred at 45°C for 3 h. TLC (PE: ethyl acetate=3:l, Rf=0.4) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the title compound 27 (26 mg, HPLC: 90%) as a light-yellow solid. Yield: 22.0%. MS- ESI: [M+l] ⁺ =494.1. *HNMR (400 MHz, DMSO): δ 9.68 (s, 1H), 7.33 (d, 1H), 7.27 - 7.16 (m, 2H), 7.13 (t, 1H), 2.73 - 2.68 (m, 4H), 2.66 (s, 6H), 2.64 (s, 3H), 2.29 (s, 3H), 2.10-2.00 (m, 4H).

Example 28: Synthesis of N-[2-(4,4-difluoro-l-piperidyl)phenyl]-5-(l-hydroxy-2-methyl -2- propyl)thiophene-2-sulfonamide

[00267] Step 1.' To a 250 mL three-necked flask was added 100 mL methanol and 2- Thiopheneacetic acid (10 g, 0.070 mol, 1.0 eq). SOCL (41.8 g, 0.352 mol, 5.0 eq) was dropped into the mixture at 0 °C. The reaction mixture was stirred at room temperature for 18 h. TLC (PE: ethyl acetate= 3: 1) showed the reaction was completed. The reaction mixture was concentrated under vacuum to afford the title compound 28A (18 g, crude) as a brown oil.

[00268] Step 2 To a 250 mL three-necked flask was added 160 mL DMF and compound 28A (8 g (crude), 0.031 mol, 1.0 eq). NaH (60% in oil) (5.13 g, 0.128 mol, 4.1 eq) was added to the mixture at 0 °C. After stirring at 0 °C for 10 min, CH3I (18.20 g, 0.128 mol, 4.1 eq) was dropped into the mixture. The reaction mixture was stirred at 0 °C for 0.5 h. TLC (PE: ethyl acetate= 10: 1) showed the reaction was completed. Water (300 mL) was added into the reaction mixture. The mixture was extracted with ethyl acetate (70 mL*3). The organic layer was dried, concentrated under vacuum and purified by column chromatography on silica to afford the title compound 28B (6 g, HPLC: 93%) as a yellow oil. Yield (two steps): 96.7%.

[00269] Step 3. To a 250 mL three-necked flask was added 50 mL dichloromethane and compound 28B (2 g, 0.011 mol, 1.0 eq) which was degassed with N2 three times. DIBAL-H (1 N in toluene) (22.8 mL, 0.023 mol, 2.1 eq) was dropped into the mixture at 0 °C. The reaction mixture was stirred at room temperature for 0.5 h. TLC (PE: ethyl acetate= 10: 1) showed the reaction was completed. Water (30 mL) and 12N HC1 (10 mL) was added into the reaction mixture at 0 °C. The water layer was extracted with dichloromethane (10 mL*3). The organic layer was dried, concentrated under vacuum to afford the title compound 28C (2.0 g, crude) as a yellow oil.

[00270] Step 4. To a 50 mL round-bottom flask was added 15 mL DMF, compound 28C (1.7 g, 0.011 mmol, 1.0 eq), imidazole (0.89 g, 0.013 mol, 1.2 eq) and TBDMS-Cl (1.64 g, 0.011 mol, 1.0 eq) which was degassed with N2 three times. The reaction mixture was stirred at room temperature for Ih. TLC (PE: ethyl acetate= 10: 1) showed the reaction was completed. Water (50 mL) was added into the reaction mixture. The water layer was extracted with ethyl acetate (20 mL*2). The organic layer was dried, concentrated under vacuum and purified by column chromatography on silica gel to afford the title compound 28D (1.6 g, HPLC: 97%) as a colorless oil. Yield (two steps): 55.2%.

[00271] Step 5 To a 50 mL round-bottom flask was added 10 mL DMF and compound 28D (1.0 g, 3.70 mmol, 1.0 eq). N-bromosuccinimide (659 mg, 3.70 mmol, 1.0 eq) was added to the mixture at 0 °C. The reaction mixture was stirred at room temperature for 16 h. Water (50 mL) was added into the reaction mixture. The water layer was extracted with ethyl acetate (20 mL*4). The organic layer was dried, concentrated under vacuum and purified by column chromatography on silica gel to afford the title compound 28E (1.0 g, HPLC: 84%) as a colorless oil. Yield: 76.9%.

[00272] Step 6 To a 100 mL round-bottom flask was added 20 mL toluene, compound 28E (1.0 g, 2.86 mmol, 1.0 eq), benzyl mercaptan (356 mg, 2.86 mmol, 1.0 eq), N,N-diisopropylethylamine (741 mg, 5.73 mmol, 2.0 eq), Xantphos (100 mg, 0.17 mmol, 0.06 eq) and Pd(dppf)C12.CHC13 (70 mg, 0.086 mmol, 0.03e q). The reaction was heated to 110 °C for 2 h with nitrogen. The reaction mixture was cooled, concentrated under vacuum and purified by column chromatography on silica gel to afford the compound 28F (770 mg, HPLC: 80%) as a colorless oil. Yield: 70%.

[00273] Step 7: To a 50 mL round-bottom flask was added 15 mL dichloromethane, 3 mL water, 0.7 mL 12N HC1 and compound 28F (770 mg, 1.96 mmol, 1.0 eq). TCCA (456 mg, 1.96 mmol, 1.0 eq) was added to the mixture with vigorous stirring at 0 - 5 °C. The reaction mixture was stirred at room temperature for 1 h. TLC (PE) showed the reaction was completed. The reaction mixture was filtered and washed with dichloromethane. The water layer was extracted with dichloromethane (5 mL*2). The organic layer was dried and concentrated under vacuum to afford the title compound 28G (0.84 g, crude) as a yellow oil.

[00274] Step 8 To a 25 mL round-bottom flask was added 5 mL pyridine, compound 28G (724 mg, 1.96 mmol, 1.5 eq) and compound 21B (278 mg, 1.31 mmol, 1.0 eq). The reaction was stirred at room temperature for 16 h. The reaction mixture was concentrated under vacuum and purified by column chromatography on silica to afford the compound 28H (110 mg, HPLC: 87%) as a yellow oil. Yield (two steps): 10.3%.

[00275] Step 9 To a 25 mL round-bottom flask was added 5 mL tetrahydrofuran and compound 28H (110 mg, 0.20 mmol, 1.0 eq). TBAF (132 mg, 0.50 mmol, 2.5 eq) was added to the mixture. The reaction was stirred at 35 °C for 2 h. The mixture was concentrated under vacuum and purified by column chromatography on silica gel to afford the title compound 28 (15 mg, HPLC: 97.9%). Yield: 17.2%. Overall Yield: 0.5%. MS-ESI: [M-l] " = 429.0.

[00276] ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.36 (s, 1H), 7.69 (d, 1H), 7.44 (d, 1H), 7.28 - 7.23 (m, 1H), 7.21 (d, 1H), 7.15 (t, 1H), 6.82 (d, 1H), 3.55 (s, 2H), 2.95 - 2.72 (m, 4H), 2.32 - 2.10 (m, 4H), 1.35 (s, 6H).

Example 29: Synthesis of 5-(tert-butyl)-N-[2-(4,4-difhioro-l-piperidyl)phenyl]thiophe ne-2- sulfonamide

[00277] Step 1. To a 500 mL three necked flask was added 100 mL dichloromethane and A1CL (40 g, 0.3 mol, 1 eq) at -78 °C, then a solution of thiophene (25.2 g, 0.3 mol, 2.0 eq) and t-butyl bromide (41.1 g, 0.3 mol, 1 eq) were added drop wise to the mixture, the reaction mixture was stirred for 1 h under -78 °C. Then it was allowed to warm to 20 °C. TLC (PE) showed a new point. The reaction mixture was diluted with dichloromethane (100 mL), and washed with water, 5% NaOH (50 mL), and water (50 mL). The organic layer was dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the compound 29A (15 g, HPLC: 95%) as a light oil. Yield: 35.7%.

[00278] Step 2 To a 100 mL three necked flask was added 25 mL DMF and compound 29A (5 g, 30 mmol, 1.0 eq), then N-chlorosuccinimide (5.7 g, 30 mmol, 1.0 eq) was added to the reaction mixture in portion under 0-5°C, TLC (PE) showed a new point. The mixture was cooled and concentrated under vacuum and the resulting product was purified by column chromatography on silica gel to afford the title compound 29B (6.0 g, HPLC: 95%) as a light oil. Yield: 91.5%.

[00279] Step 3 To a 100 mL round-bottom flask was added 20 mL toluene, compound 29B (6.0 g, 30 mmol, 1.0 eq), benzyl mercaptan (4.05 g, 28.5 mmol, 0.095 eq), N,N-diisopropylethylamine (7.4 g, 60 mmol, 2.0 eq), Pd(dppf)cl2 (114 mg, 0.13 mmol, 0.025 eq), 4,5-Bis(diphenylphosphino)- 9,9-dimethylxanthene (320 mg, 0.52 mmol, 0.1 eq), the mixture was heat to reflux in the nitrogen atmosphere overnight, TLC (PE) showed the reaction was completed and the mixture was cooled and concentrated under vacuum and the resulting product was purified by column chromatography on silica gel to afford the compound 29C (8.0 g, HPLC: 90%) as an oil. Yield: 90.5%.

[00280] Step 4 To a 100 mL three necked flask was added 40 mL glacial acetic acid, 5 mL water and compound 29C (2.62 g, 0.01 mol, 1.0 eq), then N-chlorosuccinimide (5.3 g, 0.04 mol, 4.0 eq) was added to the reaction mixture in portion under 10-15 °C, then the reaction mixture was stirred for 1 h under 10-15 °C, TLC (PE) showed the reaction was completed. The mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the compound 29D (350 mg, HPLC: 95%) as an off-white solid. Yield: 46.6%.

[00281] Step 5 To a 50 mL round-bottom flask was added 5 mL pyridine, compound 29D (350 mg, 1.5 mmol, 1.0 eq) and compound 21B (212 mg, 1 mmol, 1.0 eq), the reaction was stirred at 45 °C for 2 h. TLC (PE: ethyl acetate=10:l) showed the reaction was completed, the mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel to afford the compound 29 (150 mg, HPLC: 97%) as an off-white solid. Yield: 36.2% Overall Yield: 5.0%. MS-ESI: [M+l] ⁺ = 415.5. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.33 (s, 1H), 7.70 (d, 1H), 7.38 (d, 1H), 7.27 - 7.12 (m, 3H), 6.75 (d, 1H), 2.95 - 2.72 (m, 4H), 2.32 - 2.10 (m, 4H), 1.35 (s, 9H).

Example 30: Synthesis of N-[2-(4,4-Difluoro-l-piperidyl)phenyl]-5-(2-methoxy-2-propyl ) thiophene-2-sulfonamide

[00282] Step 1 : To a 500 mL three necked flask was added 100 mL THF and thiophene (17.0 g, 0.2 mol, 1.0 eq) at -78 °C, then n-Butyllithium (80 mL, 0.2 mol, 1 eq) were added dropwise to the mixture, the reaction mixture was stirred for 1 h under -78 °C. Then Acetone (16.8 g, 0.2 mol, 1 eq) were added dropwise to the mixture. TLC (petroleum ether: EA=10:l, Rf= 0.4) showed the reaction was having a new point. The reaction was quenched with NH4C1 (100 mL), and extracted with EA (100*3). The layer was dried over Na ₂ SO ₄ and filtered. The mixture was cooled and concentrated in vacuum and e resulting product was purified by column chromatography on silica gel to yield the product 30A (21 g) as a light oil. Yield: 84.6%.

[00283] Step 2 To a 100 mL three necked flask was added 50 mL DMF and compound 30A (21 g, 0.17 mol, 1.0 eq), then NaH (7 g, 6.8 mmol, 1.05 eq) was added to the reaction mixture in batches under 0-5 °C, Then Mel (36.21 g, 0.225 mol, 1.5 eq) were added dropwise to the mixture. TLC (petroleum ether: EA=10:l, Rf= 0.7) showed the reaction was having a new point. The reaction was quenched with H ₂ O (10 mL), and extracted with EA (100*3). The layer was dried over Na ₂ SO ₄ and filtered. The mixture was cooled and concentrated in vacuum and the resulting product was purified by column chromatography on silica gel to yield the title product 30B (22 g) as a light oil. Yield: 94.1%.

[00284] Step 3. To a 100 mL three necked flask was added 25 mL DMF and compound 30B (6 g, 40 mmol, 1.0 eq), then NCS (7.6 g, 40 mmol, 1.0 eq) was added to the reaction mixture in batches under 0-5 °C, TLC (petroleum ether, Rf= 09) showed the reaction was having a new point. The mixture was cooled and concentrated in vacuum and the resulting product was purified by column chromatography on silica gel to yield the title product 30C (7.0 g) as a light oil. Yield: 75.0%.

[00285] Step 4. To a 100 mL round-bottom flask was added 20 mL toluene, compound 30C (7.0 g, 30 mmol, 1.0 eq), benzyl mercaptan (4.05 g, 28.5 mmol, 0.095 eq), DIPEA (7.4 g, 60 mmol, 2.0 eq), Pd(dppf)C12 (114 mg, 0.13 mmol, 0.025 eq), 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene (320 mg, 0.52 mmol, 0.1 eq), the mixture was heat to reflux in a N2 atmosphere for overnight, TLC (petroleum ether, Rf= 0.8) showed the reaction was complete, and the mixture was cooled and concentrated in vacuum and the resulting product was purified by column chromatography on silica gel to yield the product 30D (6 g) as an oil. Yield: 66.6%.

[00286] Step 5 To a 100 mL three necked flask was added 40 mL glacial acetic acid, 5 mL water and compound 30D (2.78 g, 0.01 mol, 1.0 eq), then NCS (5.3 g, 0.04 mol, 4.0 eq) was added to the reaction mixture in batches under 10-15 °C, then the reaction mixture was stirred for 1 h under 10-15 °C, TLC (petroleum ether, Rf= 0.3) showed the reaction was complete. The mixture was concentrated in vacuum and the resulting product was purified by column chromatography on silica gel to yield the product 30E (1.35 g) as an off-white solid. Yield: 50.0%.

[00287] Step 6 To a 50 mL round-bottom flask was added 5 mL pyridine, compound 30E (390 mg, 1.5 mmol, 1.0 eq) and compound 21B (212 mg, 1 mmol, 1.0 eq), the reaction was stirred at 45 °C for 2 h. TLC (petroleum ether: EA=10:l, Rf= 0.3) showed the reaction was complete. The mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the product 30 (50 mg, HPLC: 97%) as an off-white solid. Yield: 7.75%. Overall Yield: 1.56%. MS-ESI: [M-l]- = 429.0. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.08 (s, 1H), 7.69 (d, 1H), 7.41 (d, 1H), 7.26 - 7.16 (m, 2H), 7.15 - 7.09 (m, 1H), 6.81 (d, 1H), 3.11 (s, 3H), 2.81-2.73 (m, 4H), 2.22 - 2.08 (m, 4H), 1.54 (s, 6H).

Example 31: Synthesis of N2,N4,N4-Trimethyl-N2-[2-(l-piperidinyl)phenyl]thiophene-2,4 - disulfonamide [00288] Step 1. To a 10 ml single-mouth round bottom flask was added compound 12 (100 mg, 0.2328 mmol, 1.0 eq), 5ml DMF, 96mg K ₂ CO ₃ (0.6983 mmol, 3.0 eq). The reaction mixture was stirred under 25 °C for 1 h. 0.5 ml Mel was added. The solution was stirred under 25 °C for 2 h. TLC showed the reaction was completed. 20 ml water was added. The solution was extracted by MTBE (30 ml*3). The organic phase was washed with brine (30 ml*2), dried over Na ₂ SO ₄ , concentrated under vacuum and purified by column chromatography on silica gel eluted with PE:EA=3:1 to afford compound 31 (44 mg) as a solid. Yield=42.6%. MS-ESI: [M+l] ⁺ = 444.10. ‘HNMR (400 MHz, CDCI ₃ ): δ 8.14 (s, 1H), 7.72 (d, 1H), 7.31 (s, 1H), 7.17 (brs, 1H), 6.96 (t, 2H), 3.33 (s, 3H), 3.02 (s, 4H), 2.80 (s, 6H), 1.73 (s, 3H), 1.59 (d, 3H).

Example 32: Synthesis of N2,N2,N5-Trimethyl-N5-[2-(l-piperidinyl)phenyl]thiophene-2,5 - disulfonamide

[00289] Step 1. To a 50 mL round-bottom flask was added 10 mL DMF, compound 4 (185 mg, 0.43 mmol, 1.0 eq) and potassium carbonate (179 mg, 1.3 mmol, 3.0 eq). The reaction mixture was stirred 1 h at room temperature. CH ₃ I (184 mg, 1.3 mmol, 3.0 eq) was dropped into the stirred mixture at room temperature for 2 h. HPLC showed the reaction was completed. The mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title product 32 (100 mg) as a white solid. Yield: 52%. MS-ESI: [MT1] ⁺ = 444.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 7.57 (d, 1H), 7.53 (d, 1H), 7.30 (s, 1H), 7.15 (s, 1H), 6.96 (t, 2H), 3.34 (s, 3H), 3.01 (s, 4H), 2.84 (s, 6H), 1.73 (s, 4H), 1.65 - 1.47 (m, 2H).

Example 33: Synthesis of N2,N2-Dimethyl-N5-[2-(l-piperidinyl)phenyl]thiazole-2,5- disulfonamide

[00290] Step 1. To a 25 mL round-bottom flask was added 1 mL pyridine, compound 4F (151 mg, 0.86 mmol, 1.0 q) and compound 8E (300 mg, 1.03 mmol, 1.2 eq). The reaction mixture was stirred at RT for 1 h. TLC (PE: EA= 5: 1) showed the reaction was completed. The reaction mixture was concentrated in vacuum and purified by column chromatography on silica gel eluted with PE: EA= 3: 1 to afford the mixture of compound 33 (29 mg, HPLC: 97.2%). Yield: 7.9%. MS-ESE [M- 1]’=428.9. ¹ H NMR (400 MHz, DMSO): δ 9.81 (brs, 1H), 8.47 (s, 1H), 7.35 - 7.26 (m, 1H), 7.26 - 7.17 (m, 2H), 7.17 - 7.07 (m, 1H), 2.86 (s, 6H), 2.71 - 2.58 (m, 4H), 1.66 - 1.51 (m, 4H), 1.51 - 1.38 (m, 2H).

Example 34: Synthesis of N,N-Dimethyl-5-[[l-[2-(l-piperidinyl)phenyl] ethyl] sulfonyl] thiophene-3-sulfonamide

[00291] Step 1. To a stirred solution of compound 2 ’-Fluoro acetophenone (30 g, 0.22 mol, 1 eq) and piperidine (19.6 g, 0.23 mol, 1.05 eq) in DMSO (200 mL) was added K ₂ CO ₃ (45.5 g, 0.33 mol, 1.5 eq). The reaction was stirred at 100 °C for 16 h. TLC showed the reaction was completed. The mixture was cooled to room temperature and poured into water (1 L), extracted with ethyl acetate (400 mL*4). The combined organic layers were washed with brine (300 mLX3), dried with Na ₂ SO ₄ , filtered and concentrated in vacuum to give compound 34A (42 g, Y=94.2%) as a yellow oil, which was used directly for the next step without further purification.

[00292] Step 2'. To a mixture of compound 34A (25 g, 0.12 mol, 1 eq) in methanol (250 mL) was added NaBIL (9.4 g, 0.24 mol, 2 eq) in portions at 0 °C. The reaction was stirred for another 2hrs at the same temperature. TLC showed the reaction was completed. The reaction was poured into icewater (800 mL) and extracted with ethyl acetate (500 mL*2). The combined organic layers were washed with brine, dried with Na ₂ SO ₄ , filtered and concentrated in vacuum to give compound 34B (26 g, Y=100%) as a light-yellow oil, which was used directly for the next step without further purification.

[00293] Step 3: To a mixture of compound 34B (0.5 g, 2.4 mmol, 1 eq) and TEA (320 mg, 3.2 mmol, 1.3e q) in DCM (10 mL) was added MsCl (370 mg, 3.2 mmol, 1.3 eq) drop-wise at 0 °C. The reaction was stirred for another 2 h at the same temperature. TLC showed the reaction was completed. The reaction was concentrated in vacuum at 10 °C and the residue compound 34C (1.1 g) was used directly for the next step without further purification.

[00294] Step 4. To a stirred solution of compound 12F (290 mg, 1 mmol, 1 eq) in toluene (20 mL) was added PPhs(786 mg, 3 mmol, 3 eq) at 0 °C. The reaction was allowed to warm to room temperature and stirred for another 2 h. TLC showed the reaction was completed. To the reaction was added H ₂ O (3 mL) and the reaction was stirred for another 10 min. The resulting mixture was concentrated in vacuum and the residue compound 34D (1.3 g) was used directly for the next step without further purification.

[00295] Step 5: To a mixture of crude compound 34D (1.3 g, 1 mmol, 1 eq) and crude compound 34C (1.1 g, 2.4 mmol, 2.4 eq) in DMF (15mL) was added Cs ₂ CO ₃ (l g, 3 mmol, 3 eq). The reaction was heated at 90 °C for 1 h. LCMS showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate (100 mL*2). The combined organic layers were washed with brine (100 mL*3), dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0-20% of ethyl acetate in petroleum ether) to afford compound 34E (430 mg, Y=100%) as a light-yellow oil. [00296] Step 6: To a stirred solution of compound 34E (400 mg, 1 mmol, 1 eq) in THF/MeOH (12 mL/8 mL) was added a solution of oxone (860 mg, 1.4 mmol, 1.4 eq) in H ₂ O (6 mL) drop-wise at 0 °C. The reaction was allowed to warm to room temperature and stirred for another 3 h. TLC showed the reaction was completed. The mixture was poured into water (100 mL), extracted with ethyl acetate (50 mL*3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0-35% of ethyl acetate in petroleum ether) to afford compound 34 (100 mg, Y=23%) as a white solid. Overall Yield: 21.7%. MS-ESI: [M+l] ⁺ = 443.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.01 (s, 1H), 7.71 (d, J = 7.1 Hz, 1H), 7.43 - 7.29 (m, 2H), 7.23 (t, J = 7.3 Hz, 1H), 7.02 (d, J = 6.6 Hz, 1H), 5.40 (s, 1H), 2.71 (m, 8H), 2.31 (s, 2H), 1.87 (d, J = 7.2 Hz, 3H), 1.83 - 1.41 (m, 6H).

Example 35: Synthesis of N,N-Dimethyl-5-[[l-[2-(l-piperidinyl)phenyl] ethyl] sulfonyl] thiophene-2-sulfonamide

[00297] Step 1. To a stirred solution of compound 4D (290 mg, 1 mmol, 1 eq) in toluene (20 mL) was added PPhs (786 mg, 3 mmol, 3 eq) at 0 °C. The reaction was allowed to warm to room temperature and stirred for another 2 h. TLC showed the reaction was completed. To the reaction was added H ₂ O (3 mL) and the reaction was stirred for another 10 min. The resulting mixture was concentrated in vacuum and the residue compound 35A (1.3 g) was used directly for the next step without further purification.

[00298] Step 2\ To a mixture of crude compound 35A (1.3 g, 1 mmol, 1 eq) and compound 34C (1.1 g, 2.4 mmol, 2.4 eq) in DMF (15 mL) was added CS ₂ CO ₃ (1 g, 3 mmol, 3 eq). The reaction was heated at 90 °C for 1 h. LCMS showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate (100 mL*2). The combined organic layers were washed with brine (100 mL*3), dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0-20% of ethyl acetate in petroleum ether) to afford compound 35B (410 mg, Y=100%) as a light-yellow oil.

[00299] Step 3: To a stirred solution of compound 35B (400 mg, 1 mmol, 1 eq) in THF/MeOH (12 mL/8 mL) was added a solution of oxone (860 mg, 1.4 mmol, 1.4 eq) in H ₂ O (6 mL) drop-wise at 0 °C. The reaction was allowed to warm to room temperature and stirred for another 3 h. TLC showed the reaction was completed. The mixture was poured into water (100 mL), extracted with ethyl acetate (50 mL*3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0-30% of ethyl acetate in petroleum ether) to afford compound 35 (110 mg, Y=25%) as a white solid. Overall Yield: 25%. MS-ESI: [M+l] ⁺ = 443.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 7.71 (d, J = 7.6

, 1H), 7.34 - 7.24 (m, 4H), 7.02 (s, 1H), 5.42 (s, 1H),2.74 (s, 8H), 2.34 (s, 2H), 1.87 (d, J = 7.2

, 3H), 1.71 - 1.45 (m, 6H).

Example 36: Synthesis of N-[2-(l-Piperidinyl)phenyl]-4-(trifluoromethyl)thiophene-2- sulfonamide

[00300] Step 1. To a mixture of compound 3-(Trifluoromethyl)thiophene (400 mg, 2.6 mmol, 1 eq) in HOAc (10 mL) was added NBS (1.4 g, 7.9 mmol, 3 eq). The reaction was heated at 100 °C for 18 h. GC showed the reaction was mainly completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with pentane (100 mL*3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue compound 36A (590 mg, Y=98.7%) was used directly for the next step without further purification.

[00301] Step 2'. To a stirred solution of compound 36A (590 mg, 2.55 mmol, 1 eq), benzyl mercaptan (316 mg, 2.55 mmol, 1 eq) and DIPEA (658 mg, 5.1 mmol, 2 eq) in toluene (20 mL) was added PdC12(dppf)CH ₂ C1 ₂ (64 mg, 77 umol, 3% eq) and Xantphos (88 mg, 153 umol, 6% eq). The reaction was degassed with nitrogen and heated at 100 °C for 2 h. TLC showed the reaction was completed. The mixture was cooled to room temperature and concentrated in vacuum. The residue was purification by flash chromatography on silica gel eluted PE to give compound 36B (360 mg, Y=51.5%) as a light-yellow oil.

[00302] Step 3 : To a stirred solution of compound 36B (360 mg, 1.56 mmol, 1 eq) in HOAc/DCM/H ₂ O (10 mL/2 mL/3 mL) was added NCS (832 mg, 6.23 mmol, 4 eq) at 0 °C. The reaction was allowed to warm to room temperature and stirred for another 2 h. TLC showed the reaction was completed. The mixture was concentrated in vacuum. The residue was purification by flash chromatography on silica gel eluted 0-5% of EA in PE to give compound 36C (80 mg, Y=20.5%) as a colorless oil. [00303] Step 4. To a stirred solution of compound 36C (80 mg, 0.32 mmol, 1.2 eq) and compound 4F (47 mg, 0.27 mmol, 1 eq) in DCM was added TEA (68 mg, 0.68 mmol, 2.5 eq) at 0 °C _O The reaction was allowed to warm to room temperature and stirred overnight. TLC showed the reaction was completed. The reaction was concentrated in vacuum and the residue was purified by flash chromatography on silica gel (0-15% of ethyl acetate in petroleum ether) to afford compound 36 (10 mg, Y=4.9%) as an oil. Overall Yield: 0.5%. MS-ESI: [M+l] ⁺ = 391.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.33 (b, 1H), 7.84 (s, 1H), 7.66-7.65 (m, 2H), 7.21 - 7.11 (m, 3H), 2.52 (b, 4H), 1.67 (b, 4H), 1.28 (b, 2H).

Example 37: Synthesis of N,N-Dimethyl-5-[[2-(l-piperidinyl)benzyl]sulfonyl]thiophene- 3- sulfonamide

[00304] Step 1. To a 2 L three necked flask was added 500 mL DMSO, 2-Fluorobenzaldehyde (75 g, 0.604 mol, 1.0 eq), piperidine (54.03 g, 0.635 mol, 1.05 eq) and potassium carbonate (125.3 g, 0.906 mol, 1.5 eq). The reaction mixture was stirred overnight at 100 °C. HPLC showed the reaction was completed. The mixture was cooled to room temperature and poured into water (1.5 L), and the reaction mixture was partitioned between ethyl acetate and water. The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated to afford the title compound 37A (99 g, GC: 97%.) as a yellow liquid. Yield: 87%.

[00305] Step 2'. To a 500 mL three necked flask was added 180 mL methanol and compound 37A (17.4 g, 91.9 mmol, 1.0 eq), which was stirred at 0°C for 10 min. NaBFL (6.95 g, 183.9 mmol, 2.0 eq) was added to the mixture in portions at 0 °C. Then the mixture was stirred at 0 °C for 2 h. TLC showed the reaction was completed. The reaction mixture was poured into water (800 mL). The water layer was extracted with ethyl acetate (400 mL*3). The combined organic layer was dried and concentrated in vacuum to afford the title compound 37B (17.5 g, GC:98%) as a colorless liquid.

Yield: 99%. [00306] Step 3: To a 100 mL three necked flask was added 30 mL DCM, compound 37B (318 mg, 1.66 mmol, 1.0 eq) and trimethylamine (252 mg, 2.49 mmol, 1.5 eq). MsCl (286 mg, 2.49 mmol, 1.5 eq) was added to the mixture drop-wise at 0 °C. Then the mixture was stirred at 0 °C for 2 h. TLC showed the reaction was completed. The reaction mixture was concentrated in vacuum at 5 °C and the residue compound 37C (1.3 g, crude, A179-3) was used directly for the next step at once without further purification.

[00307] Step 4: To a 100 mL three necked flask was added 20 mL toluene and compound 12F ( 200 mg, 0.69 mmol, 1.0 eq. Triphenylphosphine (544.7 mg, 2.08 mmol, 3.0 eq) was added to the mixture in portions at 0 °C. The reaction mixture was allowed to room temperature and stirred for another 2 h. TLC showed the reaction was completed. To the mixture was added water 3 mL) and the mixture was stirred for another 10 min. The solution was concentrated in vacuum and the residue compound 37D (1.3 g, crude, Al 80-2) was used directly for the next step at once without further purification.

[00308] Step 5: To a 100 mL three necked flask was added 15 mL DMF, compound 37D (1.3 g, 0.69 mmol, 1.0 eq), compound 37C (1.3 g, 1.66 mmol, 2.4 eq) and CS ₂ CO ₃ (675 mg, 2.07 mmol, 3.0 eq). The reaction mixture was stirred overnight at 90 °C. LC-MS showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate. The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the title compound 37E (180 mg) as an oil. Yield: 51% (two steps).

[00309] Step 6. To a 100 mL three necked flask was added 9 mL THF, 6 mL MeOH and compound 37E (180 mg, 0.45 mmol, 1.0 eq), which was stirred at 0 °C for 10 min. Oxone (419 mg, 0.68 mmol, 1.5 eq) in water (4 mL) was dropped into the mixture at 0 °C. The reaction mixture was allowed to room temperature and stirred for another 3 h . TLC showed the reaction was completed. The reaction mixture was poured into water. The water layer was extracted with ethyl acetate (50mL*2). The combined organic layer was dried and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the title compound 37 (31.3 mg) as a white solid. Yield: 16%. Overall Yield: 7%. MS-ESI: [M+l] ⁺ = 429.1. ¹ H NMR (400 MHz, CDC13): 8 8.07 (s, 1H), 7.62 (dd, 1H), 7.38 (s, 2H), 7.21 (s, 1H), 7.10 (s, 1H), 4.77 (s, 2H), 2.63 (d, 10H), 1.60 (d, 6H).

Example 38: Synthesis of N,N-Dimethyl-5-[[2-(l-piperidinyl)benzyl]sulfonyl]thiophene- 2- sulfonamide

[00310] Step 1. To a 100 mL three necked flask was added 15 mL DMF, compound 35A (1.3 g, 0.69 mmol, 1.0 eq), compound 37C (1.4 g, 0.83 mmol, 1.2 eq) and CS ₂ CO ₃ (674 mg, 2.07 mmol, 3.0 eq). The reaction mixture was stirred overnight at 90 °C. LC-MS showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate (40 mL*3). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the title compound 38A (160 mg) as an oil. Yield: 45%.

[00311] Step 2. To a 100 mL three necked flask was added 9 mL THF, 6 mL MeOH and compound 38A (160 mg, 0.4 mmol, 1.0 eq), which was stirred at 0 °C for 10 min. Oxone (372 mg, 0.6 mmol, 1.5 eq) in water (4 mL) was dropped into the mixture at 0 °C. The reaction mixture was allowed to room temperature and stirred for another 3 h. TLC showed the reaction was completed. The reaction mixture was poured into water. The water layer was extracted with ethyl acetate (50 mL*2). The combined organic layer was dried and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the title compound 38 (50 mg) as a white solid. Yield: 31%. Overall Yield: 14%. MS-ESI: [M+l] ⁺ = 429.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 7.61 (d, 1H), 7.36 (d, 2H), 7.14 (d, 3H), 4.75 (s, 2H), 2.87 - 2.24 (m, 10H), 1.58 (t, 6H).

Example 39: Synthesis of 5- [[2-(4,4-Dimethyl-l-piperidinyl)benzyl] sulfonyl] -N,N- dimethylthiophene-2-sulfonamide

[00312] Step 1. To a 50 mL round-bottom flask was added 10 mL DMSO, 2-Fluorobenzaldehyde (1.19 g, 9.6 mmol, 1.0 eq), compound 11A (1.5 g, 0.01 mol, 1.05 eq) and Potassium carbonate (3.30 g, 0.024 mol, 2.5 eq) which was degassed with N2 three times. The reaction was stirred at 100 °C for 16 h. TLC (PE) showed the reaction was completed. The reaction mixture was cooled, added 50mL water and extracted with EA (20 mL*2). The organic layer was washed with brine (20 mL*3), dried and concentrated in vacuum to afford the title compound 39A (2.0 g, LCMS: 94%) as an orange oil. Yield: 96.6%.

[00313] Step 2'. To a 50 mL round-bottom flask was added 20 mL MeOH, compound 39A (2.0 g, 9.2 mmol, 1.0 eq). NaBEE (697 mg, 0.018 mol, 2.0 eq) was added to the stirred mixture at 0 °C. After stirring for 2 h at 0 °C, the reaction mixture was added 100 mL water and extracted with EA (50 mL*2). The organic layer was washed with brine (50 mL*l), dried and concentrated in vacuum to afford the title compound 39B (1.9 g, LCMS: 93%) as a yellow solid. Yield: 94.0%.

[00314] Step 3: To a 10 OmL three-necked flask was added 10 mL DCM, compound 39B (363 mg, 1.65 mmol, 1.0 eq) and TEA (218 mg, 2.16 mmol, 1.3 eq). MsCl (247 mg, 2.16 mmol, 1.3 eq) was added to the stirred mixture at 0 °C. After stirring for 2 h at 0 °C, the reaction mixture was concentrated in vacuum at 5 °C to afford the title compound 39C (crude) as a solid.

[00315] Step 4. To a 50 mL three-necked flask was added 10 mL DMF, compound 39C (crude, 1.65 mmol, 2.4 eq), compound 35A (crude, 0.69 mmol, 1.0 eq) and CS ₂ CO ₃ (675 mg, 2.07 mmol, 3.0 eq) which was degassed with N2 three times. The reaction was stirred at 90 °C for 1 h. TLC (PE: EA= 5:1) showed the reaction was completed. The reaction mixture was cooled, added 80 mL water and extracted with EA (30 mL*2). The organic layer was washed with brine (20 mL*3), dried, concentrated in vacuum and purified by column chromatography on silica gel eluted with PE: EA= 10: 1 to afford the title compound 39D (285 mg) as an orange oil. Yield: 97.3%.

[00316] Step 5: To a 50 mL round-bottom flask was added 8.5 mL THF, 5.5 mL MeOH, compound 39D (280 mg, 0.66 mmol, 1.0 eq). Oxone (568 mg, 0.92 mmol, 1.4 eq) in H ₂ O (4 mL) was dropped to the stirred mixture at 0 °C. The reaction was allowed to warm to RT and stirred for another 3 h. TLC (PE: EA= 3:1) showed the reaction was completed. The mixture was poured into water (100 mL), extracted with EA (40 mL*2). The combined organic layers were washed with brine, dried, concentrated in vacuum and purified by flash chromatography on silica gel eluted with PE: EA= 3: 1 to afford the title compound 39 (98 mg) as a white solid. Yield: 32.6%. Overall Yield: 28.8%. MS-ESE [M+l] ⁺ = 457.1. ¹ H NMR (400 MHz, CDCI ₃ ): 5 7.60 - 7.66 (m, 1H), 7.42 - 7.50 (br, 2H), 7.27 - 7.06 (m, 3H), 5.0 - 4.54 (br, 2H), 2.74 (s, 6H), 2.68 - 2.52 (m, 4H), 1.53 - 1.19 (m, 4H), 0.98 (s, 6H).

Example 40: Synthesis of 5- [[l-[2-(4, 4-Dimethyl-l-piperidinyl)phenyl] ethyl] sulfonyl] -N,N- dimethylthiophene-2-sulfonamide

[00317] Step 1. To a 50 ml single-mouth flask bottom was added 1.5 g compound 11A (10.0227 mmol, 1.0 eq), 20ml DMSO, 1.24g 2 ’-Fluoroacetophenone (9.0205 mmol, 0.9 eq), 4.15 g K ₂ CO ₃ (30.0682 mmol, 3.0 eq). The reaction mixture was heated to 100°C overnight. TLC showed little SY002282 was left but all the A181-1 was finished. The mixture was poured into 100ml water, extracted by MTBE (50 ml*2). The MTBE phase was washed by brine (50 ml*2), dried over Na ₂ SO ₄ and purified by column chromatography on silica gel eluted with PE:EA=100:l to afford 1.2 g yellow solid as compound 40A. Yield=57.5%.

[00318] Step 2\ To a 50ml single-mouth flask bottom was added 20 ml MeOH, 1.2 g compound 40A (5.1872 mmol, 1.0 eq), 0.22 g NaBEE (5.2868 mmol, 1.02 eq). The reaction mixture was stirred at 25°C for 0.5 h. TLC showed the reaction was complete. The solution was concentrated under vacuum. The obtained solution was added 30ml water and extracted by DCM (30 ml*3). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford 1.2 g yellow liquid (come to solid after put into refrigerator) as compound 40B. Yield=100%.

[00319] Step 3. To a 25 ml single-mouth flask bottom was added 10 ml DCM, 386 mg compound 40B (1.6565 mmol, 1.0 eq), 268 mg TEA (2.6504mmol, 1.6 eq). The reaction mixture was cooled to 0°C and 28 5mg (2.4848 mmol, 1.5e q). Methanesulfonyl chloride was added as drop. The reaction mixture was stirred at 0°C for 2 h. TLC showed little Al 81-3 was left. 20ml water was added to the solution and the water phase was extracted by DCM (10 ml*2). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum at 10 °C. The obtained liquid compound 40C was used to Step6 without purification.

[00320] Step 4 : To a 50 ml single-mouth flask bottom was added compound 35A (0.6902 mmol, 1.0 eq), compound 40C (1.6565 mmol, 2.4 eq), 15ml DMF, 674 mg CS ₂ CO ₃ (2.0706 mmol, 3.0 eq). The reaction mixture was heated to 90°C for 1 h. LC-MS showed the reaction was complete. The solution was poured into 30 ml water and extracted by MTBE (30 ml*2). The MTBE phase was washed with brine (30 ml*2), dried over Na ₂ SO ₄ and purified by column chromatography on silica gel eluted with PE:EA=100:l to PE:EA=10:l to afford 315 mg orange liquid as compound 40D (HPLC 95%). Yield=80.1%.

[00321] Step 5: To a 50 ml single-mouth flask bottom was added 315 mg compound 40D (0.7181 mmol, 1.0 eq), 9.5 ml THF, 7.3 ml MeOH. The solution was cooled to 0°C and 618 mg Oxone (1.0053mmol, 1.4eq) in 4.3 ml water was added as drop. The reaction mixture was stirred for 1 h. TLC showed the Al 81-5 was finished. The solution was poured into 100ml water and extracted by EA (50 ml*2). The organic phase was washed with brine, dried over Na ₂ SO ₄ , concentrated under vacuum and purified by preparative TLC (PE: EA=4:1). The obtained was intermediate state. The obtained was dissolved in THF/MeOH and added oxone overnight. TLC showed the intermediate state was all finished. The solution was poured into 100ml water and extracted by EA (50 ml*2). The organic phase was washed with brine, dried over Na2SO-i, concentrated under vacuum and purified by preparative TLC (PE: EA=4:1) to afford 19mg white solid as compound 40. Yield=5.6%. Overall Yield=2.6%. MS-ESI: [M+l] ⁺ = 471.1. ¹ H NMR (400 MHz, CDC13): δ 7.71 (d, 1H), 7.36 (d, 3H), 7.27 - 7.19 (m, 1H), 7.08 (s, 1H), 5.32 (s, 1H), 2.74 (s, 10H), 1.86 (d, 3H), 1.43 (d, 4H), 1.02 (s, 6H).

Example 41: Synthesis of 5- [[l-[2-(4, 4-Dimethyl-l-piperidinyl)phenyl] ethyl] sulfonyl] -N,N- dimethylthiophene-3-sulfonamide

[00322] Step 1. To a 50 ml single-mouth flask bottom was added compound 37D (0.5177 mmol, 1.0 eq), compound 40C (1.2942 mmol, 2.5 eq), 15 ml DMF, 506 mg CS ₂ CO ₃ (1.5531 mmol, 3.0 eq). The reaction mixture was heated to 90°C for 1 h. LC-MS showed the reaction was complete. The solution was poured into 30ml water and extracted by MTBE (30 ml*2). The MTBE phase was washed with brine (30 ml*2), dried over Na ₂ SO ₄ and purified by column chromatography on silica gel eluted with PE:EA=100:l to PE:EA=10:l to afford 130 mg orange liquid as compound 41A (crude). Yield=57.2%.

[00323] Step 2'. To a 25 ml single-mouth flask bottom was added 130 mg compound 41A (0.2964 mmol, 1.0 eq), 15 ml THF:MeOH=3:2. The mixture was cooled to 0°C and 255 mg oxone (0.4149 mmol, 1.4 eq) in 3ml water was added as drop. The reaction mixture was stirred overnight. TLC showed the reaction was complete. The solution was poured into 100 ml water and extracted by EA (50 ml*2). The organic phase was washed with brine, dried over Na ₂ SO ₄ , concentrated under vacuum and purified by preparative TLC (PE: EA=4: 1) to afford 29 mg white solid as compound 41. Yield=20.9%. Overall Yield=l 1.9%. MS-ESE [M+l] ⁺ =471.1. ¹ H NMR (400 MHz, CDCI3): δ 8.02 (s, 1H), 7.71 (d, 1H), 7.34 (d, 2H), 7.25 (s, 1H), 7.09 (s, 1H), 5.38 (s, 1H), 2.69 (s, 10H), 1.87 (d, 3H), 1.43 (d, 4H), 1.01 (s, 6H).

Example 42: Synthesis of 5- [[2-(4,4-Dimethyl-l-piperidinyl)benzyl] sulfonyl] -N,N- dimethylthiophene-3-sulfonamide

[00324] Step 1. To a 50 ml single-mouth flask bottom was added compound 37D (0.5177 mmol, 1.0 eq), compound 39C (0.6212 mmol, 1.2 eq), 15 ml DMF, 506 mg CS ₂ CO ₃ (1.5531 mmol, 3.0 eq). The reaction mixture was heated to 90°C for 1 h. LC-MS showed the reaction was complete. The solution was poured into 30ml water and extracted by MTBE (30 ml*2). The MTBE phase was washed with brine (30 ml*2), dried over Na ₂ SO ₄ and purified by column chromatography on silica gel eluted with PE:EA=100:l to PE:EA=10:l to afford 227 mg orange liquid as compound 42A (crude). Yield=100%.

[00325] Step 2'. To a 25 ml single-mouth flask bottom was added 227 mg compound 42A (0.5354 mmol, 1.0 eq), 25ml THF:MeOH=3:2. The mixture was cooled to 0°C and 460 mg oxone (0.7495 mmol, 1.4 eq) in 4ml water was added as drop. The reaction mixture was stirred overnight. TLC showed the reaction was complete. The solution was poured into 100ml water and extracted by EA (50 ml*2). The organic phase was washed with brine, dried over Na ₂ SO ₄ , concentrated under vacuum and purified by preparative TLC (PE: EA=4:1) to afford 33mg white solid as compound 42. Yield=14.5%. Overall Yield=14.5%. MS-ESE [M+l] ⁺ = 457.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.08 (s, 1H), 7.61 (dd, 1H), 7.39 (s, 2H), 7.19 (d, 2H), 4.75 (s, 2H), 2.63 (d, 10H), 1.44 (s, 4H), 1.00 (s, 6H).

Example 43: Synthesis of 5-[[l-[3-Chloro-2-(4,4-dimethyl-l-piperidinyl)phenyl] ethyl] sulfonyl] -

N,N-dimethylthiophene-2-sulfonamide

[00326] Step 1. To a stirred solution of 3'-chloro-2'-fluoroacetophenone (1 g, 5.8 mmol, 1 eq) and compound 11A (912 mg, 6.1 mmol, 1.05 eq) in DMSO (20 mL) was added K ₂ CO ₃ (2.0 g, 14.5 mol, 2.5 eq). The reaction was stirred at 90°C for 16 h. TLC showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate (150 mL*3). The combined organic layers were washed with brine (150 mL*3), dried with Na ₂ SO ₄ , filtered and concentrated in vacuum to give compound 43A (1.5 g, Yield=97%) as a yellow oil, which was used directly for the next step without further purification.

[00327] Step 2\ To a mixture of compound 43A (1.5 g, 5.65 mmol, 1 eq) in methanol (20 mL) was added NaBIL (430 mg, 11.3 mmol, 2 eq) in portions at 0°C. The reaction was stirred for another 2 h at the same temperature. TLC showed the reaction was completed. The reaction was poured into ice-water (100 mL) and extracted with ethyl acetate (50mL*2). The combined organic layers were washed with brine, dried with Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0~8% of ethyl acetate in petroleum ether) to give compound 43B (1.1 g, Yield=73%) as a light-yellow solid.

[00328] Step 3: To a mixture of compound 43B (0.5 g, 1.9 mmol, 1 eq) and tri ethylamine (240 mg, 2.4 mmol, 1.3 eq) in di chloromethane (10 mL) was added methanesulfonyl chloride (280 mg, 2.4 mmol, 1.3 eq) drop-wise at 0°C. The reaction was stirred for another 2 h at the same temperature. TLC showed the reaction was completed. The reaction was concentrated in vacuum at 0 °C and the residue compound 43C (1.1g, Yield=100%) was used directly for the next step without further purification.

[00329] Step 4. To a stirred solution of compound 4D (200 mg, 0.7 mmol, 1 eq) in toluene(10 mL) was added PPhs(542 mg, 2.1 mmol, 3 eq) at 0°C. The reaction was allowed to warm to room temperature and stirred for another 2 h. TLC showed the reaction was completed. To the reaction was added H ₂ O (3 mL) and the reaction was stirred for another 10min.The resulting mixture was concentrated in vacuum and the residue compound 35A (1.3 g, Yield=100%) was used directly for the next step without further purification.

[00330] Step 5: To a mixture of crude compound 35A (1.3 g, 0.7 mmol, 1 eq) and crude compound 43C (1.1 g, 1.9 mmol, 2.7 eq) in DMF (10 mL) was added CS ₂ CO ₃ (1 g, 3 mmol, 3 eq). The reaction was heated at 90°C for 1 h. LCMS showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate (100 mL*2). The combined organic layers were washed with brine (100 mL*3), dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0-20% of ethyl acetate in petroleum ether) to afford compound 43D (220 mg, Yield=67%) as a light-yellow oil.

[00331] Step 6. To a stirred solution of compound 43D (220 mg, 0.47 mmol, 1 eq) in THF/MeOH (6 mL/4 mL) was added a solution of oxone (372 mg, 0.61 mmol, 1.3 eq) in H ₂ O (3 mL) drop-wise at 0°C. The reaction was allowed to warm to room temperature and stirred for 16 h. TLC showed the reaction was completed. The mixture was poured into water (100mL), extracted with ethyl acetate (50 mL*3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0-25% of ethyl acetate in petroleum ether) to afford compound 43 (78 mg, HPLC=95.9%, Yield=33%) as a white solid. MS-ESI: [M+l] ⁺ = 505.0. ¹ H NMR (400 MHz, CDCI ₃ ) : 8 7.64 (dd, 1H), 7.46 (d, 1H), 7.41 (d, 1H), 7.34 (dd, 1H), 7.24 (t, 1H), 5.78 (q, 1H), 3.69-3.43 (m, 2H), 2.75 (s, 6H), 2.42 (d, 1H), 1.79-1.72 (m, 4H), 1.60-1.27 (m, 4H), 1.02 (d, 6H).

Example 44: Synthesis of 5- [[3-Chloro-2-(4,4-dimethyl-l-piperidinyl)benzyl] sulfonyl] -N,N- dimethylthiophene-2-sulfonamide

[00332] Step 1. To a 100 mL three necked flask was added 15 mL DMSO, 3-chloro-2- fluorobenzaldehyde (1.44 g, 9.1 mmol, 1.0 eq), 11A (1.5 g, 10.0 mmol, l.le q) and potassium carbonate (3.78 g, 27.3 mmol, 3.0 eq). The reaction mixture was stirred overnight at 100°C. TLC showed the reaction was completed. The mixture was cooled to room temperature and poured into water (200 mL), and the reaction mixture was partitioned between ethyl acetate and water. The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered concentrated and purified by column chromatography on silica gel to afford the compound 44A (1.5 g) as a yellow liquid. Yield: 66%.

[00333] Step 2.' To a 100 mL three necked flask was added 15mL methanol and 44A (1.5 g, 6.0 mmol, 1.0 eq), which was stirred at 0°C for 10 min. NaBIL (0.45 g, 11.9 mmol, 2.0 eq) was added to the mixture in portions at 0°C. Then the mixture was stirred at 0°C for 2 h. TLC showed the reaction was completed. The reaction mixture was poured into water (100 mL). The water layer was extracted with ethyl acetate (50 mL*3). The combined organic layer was dried and concentrated in vacuum to afford the compound 44B (1.5 g) as a colorless liquid. Yield: 99%.

[00334] Step 3: To a 100 mL three necked flask was added 50 mL dichloromethane, compound 44B (630 mg, 2.48 mmol, 1.0 eq) and triethylamine (503 mg, 4.97 mmol, 2.0 eq). Methanesulfonyl chloride (569 mg, 4.97 mmol, 2.0 eq) was added to the mixture drop-wise at 0 °C. Then the mixture was stirred at 0 °C for 3 h. TLC showed the reaction was completed. The reaction mixture was concentrated in vacuum at 5 °C and the residue 44C (1.2 g, crude) was used directly for the next step at once without further purification. [00335] Step 4. To a 100 L three necked flask was added 30 mL toluene and compound 4D (300 mg, 1.04 mmol, 1.0 eq. Triphenylphosphine (815 mg, 3.11 mmol, 3.0 eq) was added to the mixture in portions at 0°C. The reaction mixture was allowed to room temperature and stirred for another 2 h. TLC showed the reaction was completed. To the mixture was added water (3 mL) and the mixture was stirred for another 10 min. The solution was concentrated in vacuum and the residue 35A (1.8 g, crude) was used directly for the next step at once without further purification.

[00336] Step 5: To a 100 mL three necked flask was added 20 mL DMF, 35A (1.8 g, 1.04 mmol, 1.0 eq), 44C (1.4 g, 2.48 mmol, 2.4 eq) and CsCO3 (1.02 g, 3.12 mmol, 3.0 eq). The reaction mixture was stirred overnight at 90°C. LC-MS showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate (40 mL*3). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the compound 44D (395 mg) as a yellow solid. Yield: 83% (two steps).

[00337] Step 8: To a 100 mL three necked flask was added 18 mL THF, 12 mL MeOH and 44D (395 mg, 0.86 mmol, 1.0 eq), which was stirred at 0°C for 10 min. Oxone (793 mg, 1.29 mmol, 1.5 eq) in water (8 mL) was dropped into the mixture at 0°C. The reaction mixture was allowed to room temperature and stirred for another 3 h. LC-MS showed the reaction was completed. The reaction mixture was poured into water. The water layer was extracted with ethyl acetate (50 mL*2). The combined organic layer was dried and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the compound 44 (271.5 mg, HPLC: 98%) as a white solid. Yield: 64%. [M+l] ⁺ = 491.0. ¹ H NMR (400 MHz, CDCI ₃ ): δ 7.53 (d, 1H), 7.46 (d, 1H), 7.44 (dd, 1H), 7.37 (dd, 1H), 7.18 (t, 1H), 4.93 (s, 2H), 3.59 (t, 2H), 2.78 (s, 6H), 2.24 (d, 2H), 1.58-1.46 (m, 2H), 1.42 - 1.28 (m, 2H), 1.02 (d, 6H).

Example 45: Synthesis of 5- [[3-Chloro-2-(4,4-dimethyl-l-piperidinyl)benzyl] sulfonyl] -N,N- dimethylthiophene-3-sulfonamide

[00338] Step 1. To a 100 mL three necked flask was added 40 mL di chloromethane, compound 44B (422 mg, 1.66 mmol, 1.0 eq) and triethylamine (336 mg, 3.32 mmol, 2.0 eq). Methanesulfonyl chloride (381 mg, 114.56 mmol, 2.0 eq) was added to the mixture drop-wise at 0°C. Then the mixture was stirred at 0°C for 3 hours. TLC showed the reaction was completed. The reaction mixture was concentrated in vacuum at 5 °C and the residue 44C (1.2 g, crude) was used directly for the next step at once without further purification.

[00339] Step 2\ To a 100 mL three necked flask was added 20 mL toluene and compound 12F (200 mg, 0.69 mmol, 1.0 eq). Triphenylphosphine (545 mg, 2.08 mmol, 3.0 eq) was added to the mixture in portions at 0°C. The reaction mixture was allowed to room temperature and stirred for another 2 h. TLC showed the reaction was completed. To the mixture was added water (3 mL) and the mixture was stirred for another 10 min. The solution was concentrated in vacuum and the residue 34D (1 g, crude) was used directly for the next step at once without further purification.

[00340] Step 3: To a 100 mL three necked flask was added 15 mL DMF, 34D (1 g, 0.69 mmol, 1.0 eq), 44C (1.2 g, 1.66 mmol, 2.4 eq) and CsCO3 (675 mg, 2.07 mmol, 3.0 eq). The reaction mixture was stirred overnight at 90°C. LC-MS showed the reaction was completed. The mixture was cooled to room temperature and poured into water (100 mL), extracted with ethyl acetate (40 mL*3). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the compound 45A (175 mg) as a yellow solid. Yield: 55.2% (two steps). [00341] Step 4. To a 100 mL three necked flask was added 9 mL THF, 6 mL MeOH and 45A (175 mg, 0.38 mmol, 1.0 eq), which was stirred at 0°C for 10 min. Oxone (352 mg, 0.57 mmol, 1.5 eq) in water (4 mL) was dropped into the mixture at 0°C. The reaction mixture was allowed to room temperature and stirred for another 3 h. LC-MS showed the reaction was completed. The reaction mixture was poured into water. The water layer was extracted with ethyl acetate (50 mL*2). The combined organic layer was dried and concentrated in vacuum. And the residue was purified by column chromatography on silica gel to afford the title compound 45 (54.7 mg, HPLC: 97.6%) as a white solid. Yield: 31 %. [M+l] ⁺ = 491.0. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.15 (d, 1H), 7.63 (s, 1H), 7.45 (dd, 1H), 7.37 (dd, 1H), 7.18 (t, 1H), 4.95 (s, 2H), 3.60 (t, 2H), 2.74 (s, 6H), 2.25 (d, 2H), 1.54 (t, 2H), 1.36 (d, 2H), 1.02 (s, 6H).

Example 46: Synthesis of N2- [2- [4-(4-Chloro-2-fhiorophenyl)-l-piperazinyl] phenyl] -N5,N5- dimethylthiophene-2,5-disulfonamide

[00342] Step 1. To a 10 ml single-mouth flask bottom was added 1 ml Pyridine, compound 4D (207 mg, 0.71 mmol, 1.2 eq). 2-[4-(4-chloro-2-fluorophenyl)piperazinyl]phenylamine (200 mg, 0.59 mmol, 1.0 eq), The reaction mixture was stirred for 2 h at 30°C. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 46 (170 mg, HPLC: 98.6%) as a light-red solid.

Yield=50.9%. MS-ESI: [M+l] ⁺ = 559.0. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.37 (br, 1H), 7.65 (dd, 1H), 7.51 (d, 1H), 7.37 (d, 1H), 7.28 -7.22 (m, 2H), 7.18 (td, 1H), 7.13 - 7.06 (m, 2H), 6.97 (t, 1H), 3.21 (s, 4H), 2.84 (s, 4H), 2.76 (s, 6H).

Example 47: Synthesis of 5-[[l-[3-Chloro-2-(4,4-dimethyl-l-piperidinyl)phenyl]ethyl] sulfonyl] - N,N-dimethylthiophene-3-sulfonamide

[00343] Step 1. To a 50 ml single-mouth flask bottom was added 20 ml toluene, compound 12F (200 mg, 0.69 mmol, 1.0 eq). PPhs (542 mg, 2.07 mmol, 3.0 eq) was added under 5°C. The reaction mixture was stirred at 0°C for 2 h. TLC showed the reaction was complete. 2ml water was added and stirred for another 10 min. Then the solution was concentrated under vacuum. The obtained compound 34D (crude) was used to Step 3 without purification.

[00344] Step 2\ To a 25 ml single-mouth flask bottom was added 10 ml dichloromethane, compound 43B (444 mg, 1.66 mmol, 1.0 eq), triethylamine (268 mg, 2.65 mmol, 1.6 eq). The reaction mixture was cooled to 0°C and methanesulfonyl chloride (285 mg, 2.48 mmol, 1.5 eq) was added as drop. The reaction mixture was stirred at 0°C for 2 h. TLC showed little compound 43B was left. 20 ml water was added to the solution and the water phase was extracted by dichloromethane (10 ml*2). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum at 10°C. The residue compound 43C was used to step3 without purification.

[00345] Step 3: To a 50 ml single-mouth flask bottom was added compound 34D (0.69 mmol, 1.0 eq), compound 43C (1.66 mmol, 2.4 eq), 15 ml DMF, CS ₂ CO ₃ (674 mg, 2.0706 mmol, 3.0 eq). The reaction mixture was heated to 90°C overnight. LC-MS showed the reaction was complete. The solution was poured into 30ml water and extracted by methyl tert-butyl ether (30 ml*2). The methyl tert-butyl ether phase was washed with brine (30 ml*2), dried over Na ₂ SO ₄ and purified by column chromatography on silica gel eluted with petroleum ether: ethyl acetate=100:l to petroleum ether: ethyl acetate=10:l to afford compound 47A (210 mg, HPLC: 95%) as an orange solid. Yield=64.4%. [00346] Step 4. To a 50 ml single-mouth flask bottom was added compound 47A (210 mg, 0.44 mmol, 1.0 eq), 12 ml THF, 8 ml MeOH. The oxone (382 mg, 0.66 mmol, 1.5 eq) in 3 ml water was added to the reaction mixture under 0°C. The reaction mixture was stirred at 30°C overnight. TLC showed the reaction was complete. The solution was poured into 50 ml water and extracted by ethyl acetate (50 ml*2). The organic phase was washed with brine, dried over Na ₃ SO4, concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 47 (45 mg, HPLC: 97.6%) as a white solid. Yield=20.1%. MS-ESI: [M+l] ⁺ = 505.1. ¹ H NMR (400 MHz, CDC1 ₃ ): δ 8.09 (d, 1H), 7.64 (dd, 1H), 7.51 (s, 1H), 7.33 (dd, 1H), 7.23 (t, 1H), 5.78 (d, 1H), 3.56 (q, 2H), 2.69 (s, 6H), 2.41 (d, 1H), 1.74 (d, 4H), 1.56 - 1.31 (m, 4H), 1.00 (d, 6H).

Example 48: Synthesis of N-[2-(l-Piperidinyl)phenyl]-5-(trifluoromethyl) thiophene-2- sulfonamide

[00347] Step 1. 2-bromothiophene (4.9 g, 30 mmol, 1 eq), ferrocene (1.9 g, 10 mmol, 0.3 eq), DMSO (60 mL), a DMSO solution of H ₂ SO ₄ (0.5 M, 60 ml) and a DMSO solution of CF ₃ I (3 M, 30 mL, 3 eq) were charged in a 3 -necked flask. A 30% H2O2 (6 mL) was added drop- wise at the rate of 0.5 mL/min. The temperature of the mixture was thus obtained rose up to 40°C -50°C. The mixture was stirred at this temperature for 20 min. GC showed the reaction was mainly completed. The mixture was cooled to room temperature and neutralized by adding an aqueous solution of Na ₃ CO ₃ . The resulting mixture was extracted with pentane (300 mL*2). The combined organic layers were washed with brine (200 mL*3), dried over Na ₃ SO4, filtered and concentrated in vacuum. The residue compound 48A (2.0 g, Yield=29.0%) was used directly for the next step without further purification. [00348] Step 2 To a stirred solution of compound 48A (2 g, 7.7 mmol, 1 eq), benzyl mercaptan (1.1 g, 8.7 mmol, 1 eq) and N,N-diisopropylethylamine (2.2 g, 17.4 mmol, 2 eq) in toluene (30 mL) was added PdC^dppffCI Cb (212 mg, 0.26 mmol, 0.03 eq) and Xantphos (301 mg, 0.52 mol, 0.06 eq). The reaction was degassed with nitrogen and heated at 100°C for 3 h. TLC showed the reaction was completed. The mixture was cooled to room temperature and concentrated in vacuum. The residue was purification by flash chromatography on silica gel eluted petroleum ether to give compound 48B (2.0 g, Y=83.9%) as a light yellow oil. [00349] Step 3 To a stirred solution of compound 48B (2 g, 1.56 mmol, 1 eq) in acetic acid/ dichloromethane/H ₂ O (60 mL/12 mL/20 mL) was added N-chlorosuccinimide (4.6 g, 34.8 mmol, 4.7 eq) at 0°C. The reaction was allowed to warm to room temperature and stirred for another 2hrs. TLC showed the reaction was completed. The mixture was poured into water (300mL) and extracted with ethyl acetate (150 mL*2). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filter and concentrated in vacuum. The residue was purification by flash chromatography on silica gel eluted 0~5% of ethyl acetate in petroleum ether to give compound 48C (610 mg, Y=33.2%) as a colorless oil.

[00350] Step 4 To a stirred solution of compound 48C (320 mg, 1.3 mmol, 1 eq) and compound 4F (224 mg, 1.3 mmol, 1 eq) in di chloromethane was added tri ethylamine (262 mg, 2.6 mmol, 2 eq) at 0 °C _O The reaction was allowed to warm to room temperature and stirred overnight. TLC showed the reaction was completed. The reaction was concentrated in vacuum and the residue was purified by flash chromatography on silica gel (0-15% of ethyl acetate in petroleum ether) to afford compound 48 (25 mg, HPLC=99.0%, Yield=4.9%) as a white solid. MS-ESI: [M+l] ⁺ = 391.0 'H NMR (400 MHz, CDCI ₃ ): δ 8.30 (br, 1H), 7.64 (d, 1H), 7.42 (br, 1H), 7.31 (d, 1H), 7.17-7.11 (m, 3H), 2.76-2.27 (m, 4H), 1.82-1.69 (m, 6H).

Example 49: Synthesis of N5-[3-Chloro-2-(4,4-dimethyl-l-piperidinyl)phenyl]-N2,N2- dimethylthiazole-2,5-disulfonamide

[00351] Step 1. To a 10 ml single-mouth flask bottom was added 2 ml pyridine, 11C (150 mg, 0.63 mmol, 1.0 eq), 8E (219 mg, 0.75 mmol, 1.2 eq). The reaction mixture was stirred at 30°C overnight. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 49 (60 mg, HPLC 89%) as a light-yellow solid. Yield=19.4%. MS-ESI: [M+l] ⁺ = 493.0. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.26 (s, 1H), 7.58 (d, 1H), 7.20 (t, 1H), 7.14 (d, 1H), 3.73 - 3.60 (m, 2H), 2.96 (s, 6H), 2.23 (d, 2H), 1.57 (td, 2H), 1.46 (d, 2H), 1.06 (s, 6H). Example 50: Synthesis of N2,N2-Dimethyl-N5-[3-methyl-2-(l-piperidinyl)phenyl] thiophene- 2,5-disulfonamide

[00352] Step 1. To a 50 ml single-mouth flask bottom was added 20 ml DMSO, 2-fluoro-3- nitrotoluene (2.5 g, 16.13 mmol, 1.0 eq), piperidine (1.65 g, 19.35 mmol, 1.2 eq), K ₂ CO ₃ (4.45 g, 32.26 mmol, 2.0 eq). The reaction mixture was stirred at 110°C overnight. TLC showed the reaction was complete. The solution was poured into 100 ml water and extracted by methyl tert-butyl ether (30 ml*3). The methyl tert-butyl ether phase was washed with brine (30 ml*2), dried over Na ₂ SO ₄ , concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether to afford compound 50A (3 g, HPLC: 98.7%) as a red oil. Yield=84.5%.

[00353] Step 2 To a 100 ml single-mouth flask bottom was added 50 ml methanol, 50A (3 g, 13.64 mmol, 1.0 eq), 0.3g Pd/C. The reaction mixture was stirred at 30°C under H2 overnight. TLC showed the reaction was complete. The solution was filtrated, concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether: ethyl acetate=20:l to afford compound 50B (2.2 g) as a red liquid. Yield=84.9%.

[00354] Step 3. To a 10 ml single-mouth flask bottom was added compound 50B (118 mg, 0.62 mmol, 0.9 eq), 2 ml pyridine, compound 4D (200 mg, 0.69 mmol, 1.0 eq). The reaction mixture was stirred at 30°C overnight. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3 : 1) to afford compound 50 (130 mg, HPLC 96.3%) as a light-yellow solid. Yield=42.4%. MS-ESI: [M+l] ⁺ = 444.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 9.14 (br, 1H), 7.50 (d, 2H), 7.37 (d, 1H), 7.12 (t, 1H), 6.89 (d, 1H), 3.15 (t, 2H), 2.75 (s, 6H), 2.59-2.45 (m, 2H), 2.37 (s, 3H), 1.90 (d, 1H), 1.81-1.55 (m, 4H), 1.33 (ddd, 1H).

Example 52: Synthesis of N2,N2-Dimethyl-N5-[2-(4-methyl-l-piperidinyl)phenyl] thiophene- 2,5-disulfonamide

[00355] Step 1. To a 100 mL round-bottom flask was added dioxane 60 mL, l-fluoro-2- nitrobenzene (5.92 g, 42 mmol, 1.0 eq), potassium carbonate (14.5 g, 105 mmol, 2.5 eq) and 4- methylpiperidine (5 g, 50 mmol, 1.2 eq) was dropped into the stirred mixture and refluxed overnight. LC-MS showed the reaction was completed. This reaction was added water 100mL and extracted with EA (40 mL*3). The organic solution was dried over sodium sulfate, filtered and concentrated to afford the compound 52A (8 g). Yield: 86.6 %.

[00356] Step 2 To a 250 mL round-bottom flask was added methanol 80 mL, compound 52A (8 g, 36.3 mmol, 1.0 eq), Raney -Nickel catalyst (0.8 g) was dropped into the stirred mixture at room temperature for overnight. TLC showed the reaction was complete. The solution was filtered and concentrated to afford the title compound 52B (2 g). Yield: 29 %.

[00357] Step 7 To a 25 mL round-bottom flask was added 5mL pyridine, compound 52B (131 mg, 0.69 mmol, 1.0 eq) and 4D (200 mg, 0.69 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to afford the compound 52 (96.5 mg, HPLC: 99.1%) as a yellow solid. Yield: 31.5 %. [M+l] ¹ = 444.1. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.37 (br., 1H), 7.65 (d, 1H), 7.60-7.47 (m, 1H), 7.38 (d, 1H), 7.26-7.06 (m, 3H), 2.76 (s, 6H), 2.71-2.36 (m, 3H), 1.75 (d, 2H), 1.65-1.46 (m, 2H), 1.46 - 1.26 (m, 2H), 1.03 (d, 3H).

Example 53: Synthesis of N2-[3-Fluoro-2-(l-piperidinyl)phenyl]-N5,N5- dimethylthiophene-2, 5- disulfonamide

[00358] Step 1. o a stirred solution of piperidine (638 mg, 7.5 mmol, 1.2 eq) and 2,3- difluoronitrobenzene (1.0 g, 6.3 mmol, 1 eq) in acetonitrile (20 mL) was added triethylamine (1.3 g, 12.6 mmol, 2 eq). The reaction was stirred at 80°C for 16 h. LCMS showed the reaction was completed. The mixture was concentrated in vacuum and the residue was purified by flash chromatography on silica gel (0~7 % of ethyl acetate in petroleum ether) to give compound 53A (1.6 g, Y=100%) as a light-yellow oil.

[00359] Step 2\ The mixture of compound 53A (1.6 g, 6.3 mmol) and Pd/C (0.2 g) in MeOH (20 mL) was degassed with H2 three times and stirred for 16 h at 30°C. LCMS showed the reaction was completed. The mixture was filtered and the filtrate was concentrated in vacuum to give compound 53B (1.4 g, Y=100%) as a light-yellow oil.

[00360] Step 3: To a mixture of compound 53B (140 mg, 0.72 mmol, 1.05 eq) in 1 mL of pyridine was added compound 4D (200 mg, 0.69 mmol, 1 eq) at 25°C. The reaction was stirred for another 2 h at the same temperature. TLC showed the reaction was completed. The reaction mixture was diluted with ethyl acetate (100 mL) and 0.5M HC1 (100 mL), brine (100 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (0-20% of ethyl acetate in petroleum ether) to afford compound 53 (180 mg, HPLC=98.7%, Yield=58%) as a white solid. MS-ESI: [M+l] ⁺ = 448.0. ¹ H NMR (400 MHz, CDCI ₃ ): 8 8.85 (br, 1H), 7.54 (d, 1H), 7.47 (d, 1H), 7.39 (d, 1H), 7.32-7.18 (m, 1H), 6.86 (dd, 1H), 3.15-3.03 (m, 2H), 2.76 (s, 6H), 2.65-2.51 (m, 2H), 1.89-1.25 (m, 6H).

Example 54: Synthesis of N2-[2-(6-Azaspiro[2.5]octan-6-yl)phenyl]-N5,N5- dimethylthiophene- 2,5-disulfonamide

[00361] Step 1. To a 10 ml single-mouth flask bottom was added 5 ml DMSO, l-fluoro-2- nitrobenzene (430 mg, 3.05 mmol, 0.9 eq), 6-azaspiro[2.5]octane hydrochloride (0.5 g, 3.39 mmol, 1.0 eq), K ₂ CO ₃ (1.4 g, 10.16 mmol, 3.0 eq). The reaction mixture was stirred at 100°C overnight. TLC showed the reaction was complete. The solution was poured into 40ml water and extracted by methyl tert-butyl ether (30 ml*3). The methyl tert-butyl ether phase was washed with brine (30 ml*2), dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 54A (700 mg, crude) as a red oil. Yield=100%.

[00362] Step 2\ To a 50 ml single-mouth flask bottom was added 20 ml methanol, 54A (700 mg, 3.02 mmol, 1.0 eq), 0.1g Pd/C. The reaction mixture was stirred at 30°C under H2 for 2 days. TLC showed the reaction was complete. The solution was filtrated, concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether: ethyl acetate=10:l to afford compound 54B (470 mg) as a red-black oil. Two Steps Yield=76.3%.

[00363] Step 3. To a 10 ml single-mouth flask bottom was added 54B (125 mg, 0.62 mmol, 0.9 eq), 2 ml pyridine, compound 4D (200 mg, 0.69 mmol, 1.0 eq). The reaction mixture was stirred at 30°C overnight. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 54 (120 mg, HPLC: 98%) as a light-yellow solid. Yield=38.2%. MS-ESI: [M+l] ⁺ = 456.0. ¹ H NMR (400 MHz, CDCL): δ 8.45 (br, 1H), 7.67 (d, 1H), 7.51 (d, 1H), 7.36 (d, 1H), 7.26 - 7.09 (m, 3H), 2.75 (s, 6H), 2.69-2.50 (m, 4H), 1.72-1.35 (m, 4H), 0.38 (s, 4H).

Example 55: Synthesis of N2-[2-(4-Benzyl-l-piperidinyl)phenyl]-N5,N5- dimethylthiophene-2, 5- disulfonamide

[00364] Step 1. To a 100 mL round-bottom flask was added dioxane 30 mL, l-Fluoro-2- nitrobenzene (2.01 g, 14.3 mmol, 1.0 eq), potassium carbonate (4.93 g, 35.7 mmol, 2.5 eq) and 4- benzylpiperidine (3 g, 17.1 mmol, 1.2 eq) was dropped into the stirred mixture and refluxed overnight. LC-MS showed the reaction was completed. This reaction was added water 100m aLnd extracted with ethyl acetate (40 mL*3). The organic solution was dried over sodium sulfate, filtered and concentrated to afford the title compound 55A (4.2 g). Yield: 99 %.

[00365] Step 2\ To a 100mL round-bottom flask was added methanol 40 mL, compound 55A (4.2 g, 14.2 mmol, 1.0 eq), Raney-Nickel catalyst (0.5 g) was dropped into the stirred mixture at room temperature for overnight. TLC showed the reaction was complete. The solution was filtered and concentrated to afford the compound 55B (1.5 g). Yield: 40 %.

[00366] Step 3: To a 25 mL round-bottom flask was added 5 mL pyridine, compound 55B (276 mg, 1.04 mmol, 1.0 eq) and 4D (300 mg, 1.04 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title product 55 (150 mg, HPLC: 97.7%) as a yellow liquid. Yield: 28%. [M+l] ⁺ = 520.0. ¹ H NMR (400 MHz, CDCI ₃ ): δ 8.32 (br., 1H), 7.64 (d, 1H), 7.59-7.48 (m, 1H), 7.40 - 7.29 (m, 3H), 7.08-7.27 (m, 6H), 2.77 (s, 6H), 2.67-2.47 (m, 5H), 1.83 - 1.31 (m, 6H).

Example 56: Synthesis of N2-[2-(4,4-Dimethyl-l-piperidinyl)-3-fluorophenyl]-N5,N5- dimethylthiophene-2,5-disulfonamide [00367] Step 1. To a 25 ml single-mouth flask bottom was added 10ml acetonitrile, compound 11A (1 g, 6.68 mmol, 1.2 eq), 2,3 -difluoronitrobenzene (886 mg, 5.57 mmol, 1.0 eq), triethylamine (1.13 g, 11.14 mmol, 2.0 eq). The reaction mixture was stirred at 90°C overnight. TLC showed the reaction was complete. The solution was concentrated under vacuum. The obtained oil was added 40 ml water and extracted by 60 ml ethyl acetate. The ethyl acetate phase was washed by 30 ml saturated citric acid. The water phase was extracted by 30 ml ml ethyl acetate. The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 56A (1.3 g, crude). Yield=92.8%.

[00368] Step 2\ To a 100 ml single-mouth flask bottom was added 40 ml methanol, 56A (1.3 g, 0.91 mmol, 1.0 eq), 0.2 g Pd/C. The reaction mixture was stirred at 30°C under H2 overnight. TLC showed the reaction was complete. The solution was filtrated, concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether: ethyl acetate=20:l to afford compound 56B (1.193 g) as an orange liquid. Yield=100%.

[00369] Step 3. To a 10 ml single-mouth flask bottom was added 56B (138 mg, 0.62 mmol, 0.9 eq), 2 ml pyridine, 200 mg 4D (0.69 mmol, 1.0 eq). The reaction mixture was stirred at 30°C for 2 h. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 56 (90 mg, HPLC: 98%) as a white solid. Yield=30.5%. MS-ESI: [M+l] ⁺ = 476.1. ¹ H NMR (400 MHz, CDCI ₃ ): 5 8.84 (br, 1H), 7.53 (d, 1H), 7.46 (d, 1H), 7.37 (d, 1H), 7.20 (dd, 1H), 6.91 - 6.81 (m, 1H), 3.35-3.16 (m, 2H), 2.75 (s, 6H), 2.39-2.16 (m, 2H), 1.69-1.35 (m, 4H), 1.03 (s, 6H).

Example 57: Synthesis of N2-[3-Chloro-2-(6-azaspiro[2.5]octan-6-yl)phenyl]-N5,N5- dimethylthiophene-2,5-disulfonamide

[00370] Step 1. To a 10 ml single-mouth flask bottom was added 5 ml DMSO, 2-chloro-3- fluoronitrobenzene (990 mg, 5.64 mmol, 1.0 eq), 6-azaspiro[2.5]octane hydrochloride (1 g, 6.77 mmol, 1.2 eq), K ₂ CO ₃ (1.56 g, 11.29 mmol, 2.0 eq). The reaction mixture was stirred at 100°C overnight. TLC showed the reaction going to half. The reaction mixture was stirred at 110°C overnight. TLC showed the reaction was complete. The solution was poured into 50ml water and extracted by methyl tert-butyl ether (50 ml*3). The methyl tert-butyl ether phase was washed with brine (20 ml*2), dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 57A (1.42 g) as an orange solid. Yield=94.7%.

[00371] Step 2\ To a 100 ml single-mouth flask bottom was added 50 ml methanol, compound 57A (1.42 g, 5.32 mmol, 1.0 eq), 0.2 g Raney Nickel. The reaction mixture was stirred at 30°C under H2 for 2 h. TLC showed the reaction was complete. The solution was filtrated, concentrated under vacuum to afford compound 57B (1.14 g) as ayellow solid. Yield=90.5%

[00372] Step 3. To a 10 ml single-mouth flask bottom was added 57B (147 mg, 0.62 mmol, 0.9 eq), 2 ml pyridine, 4D (200 mg, 0.69 mmol, 1.0 eq). The reaction mixture was stirred at 30°C for 3 h. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 57 (120 mg, HPLC: 98%) as a light-orange solid. Yield=46%. MS-ESI: [M+l] =490.0/492.0. ¹ H NMR (400 MHz, CDCL): δ 9.04 (br, 1H), 7.61 (dd, 1H), 7.51 (d, 1H), 7.36 (d, 1H), 7.17 (t, 1H), 7.09 (dd, 1H), 3.54 (t, 2H), 2.74 (s, 6H), 2.32 (d, 2H), 2.10 (td, 2H), 0.88 (d, 2H), 0.51 - 0.27 (m, 4H).

Example 58 : N2- [2-(4,4-Dimethyl- l-piperidinyl)-3-methylphenyl] -N5,N5-dimethylthiophene- 2,5-disulfonamide

[00373] Step 1. To a 100 mL round-bottom flask was added DMSO 30 mL, 2-fluoro-3- nitrotoluene (2.5 g, 16.1 mmol, 1.0 eq) , potassium carbonate (6.7 g, 48.3 mmol, 3.0 eq) and 11A (2.9 g, 19.3 mmol, 1.2 eq) was dropped into the stirred mixture, then the mixture was stirred at 120°C overnight. LC-MS showed the reaction was completed. This reaction was added water 100m aLnd extracted with ethyl acetate (50 mL*3). The organic solution was dried over sodium sulfate, fdtered and concentrated to afford compound 58A (2.5 g) as a light-yellow solid. Yield: 62.5%. [00374] Step 2 To a 100 mL round-bottom flask was added methanol 50 mL, compound 58A (2.5 g, 10.1 mmol, 1.0 eq), Pd/C (0.5 g) was dropped into the stirred mixture at room temperature under H2 for overnight. TLC showed the reaction was completed. The solution was filtered and concentrated to afford compound 58B (1.5g) as a light-yellow solid. Yield: 68.2%.

[00375] Step 3 To a 25 mL round-bottom flask was added 5 mL pyridine, compound 58B (300 mg, 1.37 mmol, 1.0 eq) and 4D (398 mg, 1.37 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title compound 58 (294 mg, HPLC: 97.7%) as a white solid. Yield: 45.4%. Overall Yield: 19.4%. [MT1] ⁺ = 472.1. ¹ H NMR (400 MHz, CDCI ₃ ) δ 9.06 (s, 1H), 7.52 -7.46 (m, 2H), 7.34 (d,lH), 7.11 (t, 1H), 6.88 (d, 1H), 3.30 (t, 2H), 2.73 (s, 6H), 2.38 (s, 3H), 2.33-2.17 (m, 2H), 1.65-1.47 (m, 2H), 1.41 (d, 2H), 1.03 (d, 6H).

Example 59: N2-[3-Ethyl-2-(l-piperidinyl)phenyl]-N5,N5-dimethylthiophene -2,5-disulfonamide

[00376] Step 1. To a 2 L round-bottom flask was added acetic acid 800 mL and 2-ethylaniline (200 g, 1.65 mol, 1.0 eq). Then acetic anhydride (253 g, 2.48 mol, 1.5 eq) was dropped into the stirred mixture at 10°C-20°C. Then the mixture was stirred at 30°C for 0.5 hours. TLC showed the reaction was completed. This reaction was added water 2 L and extracted with di chloromethane (2 L*l). The organic solution was washed with brine (500 mL*3), dried over sodium sulfate, filtered and concentrated to afford compound 59A (220 g) as a white solid. Yield: 81.5%.

[00377] Step 2 To a 100 mL round-bottom flask was added acetic acid 800 mL, compound 59A(110 g, 0.67 mol, 1.0 eq). Fuming nitric acid (340 g, 5.40 mol, 8.0 eq) was added dropwise into the mixture at a temperature of 50°C-55°C. Then the mixture was stirred at 50°C-55°C for 0.5 h. LC- MS showed the reaction was completed. The mixture was poured into ice, and extracted with dichloromethane (1 L*2). The organic layer was washed with brine (500 mL*3), dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel to afford compound 59B (50 g, crude) as a yellow solid.

[00378] Step 3: To a 1 L round-bottom flask was added 420 mL water, compound 59B (50 g, 0.24 mol, 1.0 eq) and H2SO4 (141 g, 1.44 mol, 6.0 eq), the reaction was refluxed for overnight. LC-MS showed the reaction was completed; the pH of the mixture was adjusted to 9-10 with 2 N NaOH, and extracted with dichloromethane. The organic layer was dried over sodium sulfate, concentrated in vacuum and the residue was purified by column chromatography on silica gel to afford compound 59C (10 g, GC: 90%) as a yellow liquid. Yield: 10.0% (two steps).

[00379] Step 4. To a 100 mL round-bottom flask was added 35 mL water, aqueous HBr ( 18.3 g, 90.3 mmol, 2.5 eq) and 59C ( 6 g, 36.1 mmol, 1.0 eq). The mixture was refluxed for 1 h and then cooled to 0°C. A solution of NaNCL (2.5 g, 36.1 mmol, 1.0 eq) in water (14 mL) was added dropwise at a temperature of 0°C-10°C. The mixture was stirred for 2 h at a temperature of 10°C-20°C and cooled to 0°C, a mixture of CuBr (5.18 g, 36.1 mmol, 1.0 eq) in aqueous HBr (14.6 g, 72.2 mmol, 2.0 eq) and water was added dropwise to the solution at 0°C-10°C. The mixture was stirred at 10°C-20°C for 30 min and refluxed for overnight. GC showed the reaction was completed. The mixture was extracted with ethyl acetate (200 mL*2), the organic layer was washed with aqueous saturated NaHCOs and brine, dried over sodium sulfate, concentrated in vacuum and the residue was purified by column chromatography on silica gel to afford compound 59D(3.4 g, GC:90%) as a yellow liquid. Yield: 44.6%.

[00380] Step 5: To a 100 mL round-bottom flask was added DMSO 30 mL, 59D (3.70 g, 16.1 mmol, 1.0 eq) , potassium carbonate (5.56 g, 40.2 mmol, 2.5 eq) and piperidine (1.64 g, 19.3 mmol, 1.2 eq), then the mixture was stirred at 120°C overnight. LC-MS showed the reaction was completed. This reaction was added water 100 mL and extracted with ethyl acetate (50 mL*3). The organic solution was dried over sodium sulfate, filtered and concentrated to afford compound 59E (190 mg) as a yellow liquid. Yield: 5.0%.

[00381] Step 6: To a 100 mL round-bottom flask was added methanol 20 mL, compound 59E (190 mg, 0.8 mmol, 1.0 eq), Pd/C (0.1 g) was dropped into the stirred mixture at room temperature under H2 for overnight. TLC showed the reaction was completed. The solution was filtered and concentrated to afford compound 59F (95 mg) as a yellow liquid. Yield: 57.6%. [00382] Step 7\ To a 25 mL round-bottom flask was added 5 mL pyridine, compound 59F (95 mg, 0.46 mmol, 1.0 eq) and 4D (133 mg, 0.46 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title product 59 (75.2 mg, HPLC: 99.3%) as a white solid. Yield: 35.3%. Overall Yield: 0.037%. [MT1] ⁺ = 458.1. ¹ H NMR (400 MHz, CDCI ₃ ) δ 9.10 (s, 1H), 7.52 (dd, 1H), 7.49 (d, 1H), 7.36 (d, 1H), 7.17 (t,lH), 6.95 (dd, 1H), 3.08 (td, 2H), 2.74 (s, 6H), 2.66 (q, 2H), 2.44 (d, 2H), 1.89 (d, 1H), 1.74 (d, 2H), 1.67 - 1.54 (m, 2H), 1.41 - 1.26 (m, 1H), 1.22 (t, 3H).

Example 60: Synthesis of N2-[2-(4,4-Diethyl-l-piperidinyl)-3-methylphenyl]- N5,N5- dimethylthiophene-2,5-disulfonamide

[00383] Step 1. To a 500 ml three-neck flask bottom was added 200 ml toluene, 3-pentanone (20 g, 232 mmol, 1.0 eq), ethyl cyanoacetate (36.79 g, 325 mmol, 1.4 eq), ammonium acetate (4.78 g, 58 mmol, 0.25 eq), acetic acid (16.74 g, 279 mmol, 1.2 eq). The flask was equipped with a Dean-Stark tube. The solution was refluxed under N2 overnight. GC showed the solution was nearly complete. The solution was cooled and 300 ml MTBE was added. The organic phase was washed with water (75 ml*2), dried over Na ₂ SO ₄ and concentrated under vacuum. The residure was distilled under 0.2 torr to afford 60A (35 g) as a colorless liquid. Another 62 g 3-pentanone was used to make compound 60A and 100 g of 60A as a colorless liquid was obtained. Yield=86%.

[00384] Step 2'. To a 100 ml three-neck flask bottom was added 50 ml ethanol. Sodium (1.39 g, 61 mmol, 2.0 eq) was added with water bath. After the sodium had reacted, cyanoacetamide (5.1 g, 61 mmol, 2.0 eq) was added and the reaction mixture was heated to reflux for 15 min. The mixture was cooled and a solution of 60A (5 g, 30 mmol, 1.0 eq) in 5 ml ethanol was added as a thin stream. The reaction mixture, which was very thick, turned yellow-orange and was mildly exothermic. Stirring was continued overnight, and then the slurry was poured into 60 ml water, forming a homogeneous orange solution. Hydrochloric acid was added under ice bath to adjust pH to 2-3. The solution was stirred for 10 min under ice bath and the solid was collected by suction fdtration, washed with cold water and dried under infrared. Another 60 g 60A was used to make compound 60B and total 55 g of compound 60B was obtained as a white fluffy solid. Yield=64%.

[00385] Step 3: To a 250 ml three-neck flask was added 47 ml H2SO4, 60B (20 g, 91 mmol, 1.0 eq). The flask was equipped with a magnetic stir bar and a constant pressure dropping funnel. With stirring, 15.15 ml of water was added dropwise over a 45-min period. Sometimes solids emerged and then redissolved during this procedure. The solution was heated to reflux. After 1 h this moderated, and a clear yellow-brown solution with little foam was evident. Heating was discontinued, and after 10 min, 18.18 ml of water was added dropwise over a 35-min period to the still hot solution. Then heating was resumed, and the reaction mixture was brought to reflux overnight. The solution was cooled to 30°C and was poured into 150 ml water. The solution was fdtered. The obtained solid was triturated 3 times with ether (150 ml*3). The ether phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford 60C (6 g) as a white fluffy solid. Yield=35%.

[00386] Step 4: To a 50 ml single-mouth flask bottom was added urea (5 g, 66 mmol, 25 eq), 60C (0.5 g, 2.66 mmol, 1.0 eq). The solution was heated to 160°C for 1 h under N2. Then heating was discontinued and 30 ml water was added. After the solution was cooled to 30°C, it was extracted by ethyl acetate (15 ml*2). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum. Another 5.5 g compound 60C was used to make compound 60D. Compound 60D (total 3 g) was obtained as a white solid. Yield=56%.

[00387] Step 5: To a 100 ml three-neck flask bottom was added 20 ml THF. LiAlHi (1.01 g, 26.63 mmol, 3.0 eq) was added at 0°C. A solution of 60D (1.5 g, 8.88 mmol, 1.0 eq) in 15 ml THF was added to the reaction mixture at 0-5°C. The solution was stirred at 25°C for 1 h and then refluxed for 3 h. The solution was cooled and 1 ml water, 2 ml 10% NaOH, 1ml water was dropped to the solution under 5°C. The mixture was filtered and the solid was washed with dichloromethane. The organic phase was dried over Na ₂ SO ₄ and 30 ml 10 mol/L HCl-ether was added. The obtained solution was concentrated under vacuum to afford 60E (1.5 g) as a brown oil. It was used to next step without purification. [00388] Step 6: To a 50 ml single-mouth flask bottom was added 20 ml DMSO, 2-fluoro-3- nitrotoluene (0.92 g, 5.92 mmol, 1.0 eq), 60E (1.5 g, 8.88 mmol, 1.5 eq), K ₂ CO ₃ (2.45 g, 17.75 mmol, 3.0 eq). The reaction mixture was stirred at 120°C overnight. TLC showed the reaction was complete. The solution was poured into 200 ml water and extracted by MTBE (100 ml*3). The MTBE phase was washed with brine (50 ml*2), dried over Na ₂ SO ₄ , concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether to afford compound 60F (1 g) as an orange solid (HPLC 30%).

[00389] Step 7: To a 50 ml single-mouth flask bottom was added 25 ml Methanol, 60F (1 g, 3.62 mmol, 1.0 eq), 0.5 g Pd/C. The reaction mixture was stirred at 50°C for 6 h. TLC showed the reaction was complete. The solution was filtrated, concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether: ethyl acetate=50:l to afford compound 60G (158 mg) as an orange-red liquid. Three steps yield=7.2%.

[00390] Step 8: To a 10 ml single-mouth flask bottom was added 60G (158 mg, 0.6423 mmol, 1.0 eq), 2 ml Pyridine, 4D (167 mg, 0.5780 mmol, 0.9 eq). The reaction mixture was stirred at 25°C for 3 h. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford 130 mg orange oil. The obtained oil was purified by preparative TLC (dichloromethane) to afford compound 60 (96 mg) as a light-yellow solid. Yield=33%. Over all yield=0.26%. [M+l] ⁺ = 500.1. ¹ H NMR (400 MHz, CDCI3) 8 9.03 (s, 1H), 7.48 (dd, 2H), 7.33 (d, 1H), 7.08 (t, 1H), 6.85 (d, 1H), 3.26 (dt, 2H), 2.72 (s, 6H), 2.35 (s, 3H), 2.16 (d, 2H), 1.54 (d, 2H), 1.47 (q, 2H), 1.42 - 1.24 (m, 4H), 0.81 (dt, 6H).

Example 63: Synthesis of N2-[2-(3-Azabicyclo[3.1.1]heptan-3-yl)-3-chlorophenyl]-N5,N5 - dimethylthiophene-2,5-disulfonamide

[00391] Step 1. To a 25 ml single-mouth flask bottom was added 10 ml dioxane, l-chloro-2- fluoro-3 -nitrobenzene (657 mg, 3.74 mmol, 1.0 eq), 3 -azabicyclo [3.1.1 ]heptane hydrochloride (500 mg, 3.74 mmol, 1.0 eq), K ₂ CO ₃ (1.55 g, 11.23 mmol, 3.0 eq). The reaction mixture was stirred at 110°C for 4 h. LC-MS showed the reaction was complete. The solution was poured into 30ml water and extracted by MTBE (20 ml*3). The MTBE phase was dried over Na ₂ SO ₄ , concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether to afford compound 63A (840 mg) as a red oil. Yield=88.8%.

[00392] Step 2\ To a 25 ml single-mouth flask bottom was added 15 ml methanol, 63A (840 mg, 3.32 mmol, l.Oe q), 0.1g Raney Ni. The reaction mixture was stirred at Rt (18°C) overnight under H2. LC-MS showed the reaction was complete. The solution was filtrated, concentrated under vacuum to afford compound 63B (720mg) as a light orange liquid. Yield=97.3%.

[00393] Step 3: To a 10 ml single-mouth flask bottom was added 63B (200 mg, 0.90 mmol, 1.0 eq), 2 ml Pyridine, 4D (234 mg, 0.81 mmol, 0.9 eq). The reaction mixture was stirred at Rt (18°C) for 2 h. LC-MS showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 63 (212 mg) as a light-orange solid. Yield=55.1%. Overall Yield=47.6%. [M+l] ⁺ = 476.0. ¹ H NMR (400 MHz, CDCI ₃ ) δ 8.64 (s, 1H), 7.61 (dd, 1H), 7.55 (d, 1H), 7.39 (d, 1H), 7.21 (t, 1H), 7.16 (dd, 1H), 3.60 (d, 2H), 2.77 (m, 8H), 2.50 (t, 2H), 2.22 (brs, 2H), 1.93 (t, 1H), 1.62 (t, 1H).

Example 64: Synthesis of N2-[2-(l-Azepanyl)-3-fluorophenyl]-N5,N5- dimethylthiophene-2, 5- disulfonamide

[00394] Step 1. To a 25 ml single-mouth flask bottom was added 10 ml dioxane, 2,3- difluoronitrobenzene (1 g, 6.28 mmol, 1.0 eq), hexamethyleneimine (1.65 g, 7.54 mmol, 1.2 eq), K ₂ CO ₃ (1.73 g, 12.57 mmol, 2.0 eq). The reaction mixture was stirred at 105°C for 1 h. TLC showed the reaction was complete. The solution was poured into 40 ml water and extracted by MTBE (30 ml*3). The MTBE phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 64A (1.4 g, crude) as a red oil. It was used to next step without purification.

[00395] Step 2\ To a 100 ml single-mouth flask bottom was added 30 ml methanol, 64A (1.4 g, 6.28 mmol, 1.0 eq), 0.2 g Raney Ni. The reaction mixture was stirred at Rt (18°C) overnight under H2. TLC showed the reaction was complete. The solution was filtrated, concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether- petroleum ether: ethyl acetate=10:l to afford compound 64B (1.33 g) as a light orange liquid. Two steps yield=100%. [00396] Step 3: To a 10 ml single-mouth flask bottom was added 64B (150 mg, 0.72 mmol, 1.0 eq), 2 ml Pyridine, 4D (188 mg, 0.65 mmol, 0.9 eq). The reaction mixture was stirred at Rt (18°C) for 2 h. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 64 (152 mg) as a light-pink solid. Yield=45.8%. Overall Yield=45.8%. [M+l] ⁺ = 462.1. ¹ H NMR (400 MHz, CDCI ₃ ) 5 8.71 (s, 1H), 7.55 (d, 1H), 7.44 (d, 1H), 7.39 (d, 1H), 7.16 (td, 1H), 6.84 (ddd, 1H), 3.25-2.45 (m, 10H), 1.85-1.62 (m, 8H).

Example 65: N2-[2-(l-Azepanyl)-3-chlorophenyl]-N5,N5-dimethylthiophene-2 ,5-disulfonamide

[00397] Step 1. To a 100 mL round-bottom flask was added 1,4-dioxane 10 mL, l-chloro-2- fluoro-3 -nitrobenzene (2 g, 11.4 mmol, 1.0 eq) , potassium carbonate(3.94 g, 28.5 mmol, 2.5 eq) and hexamethyleneimine (1.24 g, 12.5 mmol, 1.1 eq) was dropped into the stirred mixture, then the mixture was refluxed overnight. TLC showed the reaction was completed. This reaction was added water 50 mL and extracted with ethyl acetate (20 mL*3). The organic solution was dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel to afford the title compound 65A (2.5 g) as a yellow liquid. Yield: 86.2%.

[00398] Step 2\ To a 100 mL round-bottom flask was added methanol 25 mL, compound 65A (2.5 g, 9.8 mmol, 1.0 eq), Raney Ni (0.25 g) was dropped into the stirred mixture at room temperature under H2 for overnight. LC-MS showed the reaction was completed. The solution was filtered and concentrated to afford the compound 65B (1.9 g) as a yellow liquid. Yield: 86.4%.

[00399] Step 3: To a 25 mL round-bottom flask was added 5 mL pyridine, compound 65B (200 mg, 0.89 mmol, 1.0 eq) and 4D (256 mg, 0.89 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title product 65 (145 mg, HPLC: 99.8%) as a white solid. Yield: 34.0%. Overall Yield: 25.3%. [M+l] ⁺ = 478.0. ¹ H NMR (400 MHz, CDCI ₃ ) δ 9.02 (s, 1H), 7.60 - 7.55 (m, 2H), 7.39 (d, 1H), 7.18 - 7.08 (m, 2H), 3.46 - 3.32 (m, 2H), 2.76 (s, 6H), 2.60 - 2.47 (m, 2H), 1.88 - 1.59 (m, 8H).

Example 66: Synthesis of N2-[3-Chloro-2-(l,l-dioxidothiomorpholino)phenyl]-N5,N5- dimethylthiophene-2,5-disulfonamide

[00400] Step 1. To a 50 ml single-mouth flask bottom was added 20 ml dioxane, l-chloro-2- fluoro-3 -nitrobenzene (2 g, 11.40 mmol, 1.0 eq), 1 , 1 -dioxi de-4-thiomorpholine (1.85 g, 13.68 mmol, 1.2 eq), K ₂ CO ₃ (3.14 g, 22.79 mmol, 2.0 eq). The reaction mixture was stirred at 110°C for 4 h. TLC showed it did not react. 20 ml DMSO was added to the solution and heated to 120 °C overnight. TLC showed the reaction was complete. The solution was poured into 300 ml water and extracted by MTBE (100 ml*3). The MTBE phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 66A (150 mg) as an orange solid. Yield=4.5%.

[00401] Step 2'. To a 25 ml single-mouth flask bottom was added 15 ml methanol, 66A (150 mg, 0.52 mmol, 1.0 eq), 0.1 g Raney Ni. The reaction mixture was stirred at 18°C overnight under H2. TLC showed the reaction was complete. The solution was filtrated and concentrated under vacuum to afford compound 66B (120 mg) as a brown solid. Yield=89.2%.

[00402] Step 3. To a 10ml single-mouth flask bottom was added 66B (120 mg, 0.46 mmol, 1.0 eq), 2 ml Pyridine, 4D (120 mg, 0.42 mmol, 0.9 eq). The reaction mixture was stirred at Rt (18°C) for 2 h. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford 71 mg (HPLC 86%) orange solid. The obtained solid was stirred with 5 ml petroleum ether: dichloromethane=l :l for 10 min. The solution was filtrated to afford compound 66 (60 mg) as a white solid. Yield=28.2%. Overall Yield= 1.1%. [M+l] ⁺ = 514.0. ¹ H NMR (400 MHz, DMSO) δ 10.13 (s, 1H), 7.73 (d, 1H),

7.68 (d, 1H), 7.38 - 7.31 (m, 1H), 7.25 (d, 2H), 3.68 (d, 2H), 3.60 (d, 2H), 3.04 (d, 2H), 2.91 (d, 2H),

2.68 (s, 6H).

Example 67 : N2- [3-Chloro-2-(4,4-dichloro- l-piperidinyl)phenyl] -N5,N5-dimethylthiophene-2,5- disulfonamide

[00403] Step 1. To a 100 mL round-bottom flask was added 1,4-dioxane 50 mL, l-chloro-2- fluoro-3 -nitrobenzene (5 g, 28.5 mmol, 1.0 eq), potassium carbonate (9.8 g, 71.2 mmol, 2.5 eq) and 4-piperidinol_(3.17 g, 31.3 mmol, 1.1 eq) was dropped into the stirred mixture, then the mixture was refluxed overnight. LC-MS showed the reaction was completed. This reaction was added water 200 mL and extracted with ethyl acetate (50 mL*3). The organic solution was dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel to afford 67A (6 g) as a yellow solid. Yield: 82.1%.

[00404] Step 2'. To a 100 mL three necked flask was added 60 mL dichloromethane and oxalyl chloride (5.93 g, 46.7 mmol, 2.0 eq). The mixture was cooled to -80°C and treated drop wise with a solution of DMSO (9.13 g, 116.9 mmol, 5.0 eq) in di chloromethane (25 mL). The mixture was stirred at -80°C for 15 min. Then the mixture was treated dropwise with a solution of 67A (6 g, 23.4 mmol, 1.0 eq) in di chloromethane (25 mL). The mixture was stirred at -70°C for 1.5 h. And the mixture was dropwise with a solution of TEA (11.83 g, 116.9 mmol, 5.0 eq) in dichloromethane (25 mL). The mixture was slowly warmed to room temperature and stirred overnight. TLC showed the reaction was completed. The reaction mixture was partitioned between dichloromethane and water. The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered, concentrated, and purified on silica gel to afford 67B (5.9 g) as a yellow solid. Yield: 99%.

[00405] Step 3. To a 250 mL three necked flask was added 60 mL toluene, 67B (5.9 g, 23.2 mmol, 1.0 eq) and PCI5 (12.1 g, 57.9 mmol, 2.5 eq). The reaction was stirred at room temperature for 2 h. TLC showed the reaction was completed. The mixture was adjusted with 2 N NaOH to pH=10- 11. The mixture was extracted with methyl tert-butyl ether (100 mL*3). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered, concentrated, and purified on silica gel to afford 67C (3.2 g) as a yellow solid. Yield: 44.4%.

[00406] Step 4. To a 100 mL round-bottom flask was added methanol 50mL, compound 67C (1.7 g, 5.5 mmol, 1.0 eq), Raney Ni (0.2 g) was dropped into the stirred mixture at room temperature under H2 for 1 h. TLC showed the reaction was completed. The solution was filtered and concentrated to afford the compound 67D (1.1 g) as ayellow solid. Yield: 73.3%.

[00407] Step 5: To a 25 mL round-bottom flask was added 10 mL pyridine, compound 67D (200 mg, 0.72 mmol, 1.0 eq) and 4D (207 mg, 0.72 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title product 67 (141 mg, HPLC: 95.4%) as ayellow solid. Yield: 37.0%. Overall Yield: 9.8%. [M-l]’ =529.7. ¹ H NMR (400 MHz, CDCI ₃ ) δ 8.51 (s, 1H), 7.60 (dd, 1H), 7.53 (d, 1H), 7.40 (d,lH), 7.21 (t, 1H), 7.12 (dd, 1H), 3.89 (td, 2H), 2.79 (s, 6H), 2.57 (d, 2H), 2.50 - 2.39 (m, 2H), 2.34 (dd, 2H).

Example 68 : N2- [3-Chloro-2-(4-chloro- l-piperidinyl)phenyl] -N5,N5-dimethylthiophene-2,5- disulfonamide

[00408] Step 1. To a 100 mL three necked flask was added 30 mL chloroform, 67A (2.5 g, 9.7 mmol, 1.0 eq) and PCI5 (2.4 g, 11.7 mmol, 1.2 eq). The reaction was stirred at room temperature for 2 h. TLC showed the reaction was completed. The mixture was adjusted with 2 N NaOH to pH=10-l 1. The mixture was extracted with dichloromethane (100 mL*3). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered, concentrated, and purified on silica gel to afford 68A (1.2 g) as a yellow liquid. Yield: 44.9%.

[00409] Step 2'. To a 100 mL round-bottom flask was added methanol 20 mL, compound 68A (1.2 g, 4.4 mmol, 1.0 eq), Raney Ni (0.2 g) was dropped into the stirred mixture at room temperature under H2 for 1 h. TLC showed the reaction was completed. The solution was filtered, concentrated and purified on silica gel to afford the compound 68B (0.7 g) as a yellow liquid. Yield: 65.4%.

[00410] Step 3: To a 25 mL round-bottom flask was added 10 mL pyridine, compound 68B (186 mg, 0.7 mmol, 1.1 eq) and 4D (200 mg, 0.7 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title product 68 (96 mg, HPLC: 98.5%) as a white solid. Yield: 25.4%. Overall Yield: 5.3%. [M+l] ⁺ = 498.0. ¹ H NMR (400 MHz, MeOD) δ 7.71-7.46 (m, 3H), 7.31-7.16 (m, 2H), 4.53 (s, 0.5H), 4.01 (s, 0.5H), 3.82 (d, 1H), 3.50 (t, 1H), 2.71 (s, 6H), 2.37 (d, 1H), 2.29-2.11 (m, 3H), 2.03-1.87 (m, 2H).

Example 71: Synthesis of N2-[3-Chloro-2-(4-ethyl-4-methyl-l-piperidinyl)phenyl]- N5,N5- dimethylthiophene-2,5-disulfonamide

[00411] Step 1. To a 250 ml three-necked flask bottom was added 70 ml THF, LiAlIL (3.68 g, 96.77 mmol, 3.0 eq). A solution of 4-ethyl-4-methylpiperidine-2, 6-dione (5 g, 32.26 mmol, 1.0 eq) in 80 ml THF was added to the bottom under 10°C. The reaction mixture was stirred for 1 h at 15°C and then reflux for 3 h. TLC showed the reaction was complete. The solution was cooled to 0°C and 3.68 ml water, 7.36 ml 10% NaOH, 3.68 ml water was added as turn at 0°C. The solution was filtered; the solid was soaked with 150 ml di chloromethane and then filtered. All the organic phase was added excess HCl/EtOH and then concentrated under vacuum. The obtained oil was stirred with 50ml petroleum ether and then filtered to afford 71A (6.3 g) as a pink solid. Yield=100%.

[00412] Step 2'. To a 25 ml single-mouth flask bottom was added 10 ml dioxane, 71A (1 g, 6.13 mmol, 1.5 eq), SY015562 (0.72 g, 4.09 mmol, 1.0 eq), K ₂ CO ₃ (1.69 g, 12.27 mmol, 3.0 eq). The reaction mixture was stirred at 110°C for 3 h. TLC showed the reaction was complete. The solution was cooled, poured into 30 ml water and extracted by MTBE (30 ml*3). The organic phase was dried over Na ₂ SO ₄ , concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether to afford 71B (0.93 g) as an orange solid. Yield=80.4%.

[00413] Step 3. To a 50 ml single-mouth flask bottom was added 50 ml methanol, 71B (0.93 g, 3.29 mmol, 1.0 eq), 0.1 g Raney Ni. The reaction mixture was stirred at 15°C overnight under Th. TLC showed the reaction was complete. The solution was filtered and the organic phase was concentrated under vacuum to afford 71C (870 mg) as an orange liquid. Yield=100%.

[00414] Step 4. To a 10 ml single-mouth flask bottom was added 2 ml pyridine, 71C (200 mg, 0.79 mmol, 1.2 eq), 4D (190 mg, 0.66 mmol, 1.0 eq). The reaction mixture was stirred for 2 h. TLC showed the reaction was complete. The solution was concentrated under vacuum and purified by preparative TLC (petroleum ether: ethyl acetate=3:l) to afford compound 71 (155 mg) as a light pink solid. Yield=46.4%. Over all yield=37.3%. [M+l] ⁺ = 506.1/508.1. ¹ H NMR (400 MHz, CDCI ₃ ) δ 8.99 (brs, 1H), 7.61 (dd, 1H), 7.51 (dd, 1H), 7.38 (d, 1H), 7.17 (t, 1H), 7.09 (dt, 1H), 3.65-3.55 (m, 2H), 2.75 (s, 6H), 2.12 (dd, 2H), 1.57-1.43 (m, 5H), 1.38 - 1.25 (m, 2H), 0.97 (d, 3H), 0.90 (t, 2H).

Example 72 : N2- [2-(4-Ethyl-4-methyl- l-piperidinyl)-3-methylphenyl] -N5,N5- dimethylthiophene-2,5-disulfonamide

[00415] Step 1. To a 100 mL round-bottom flask was added DMSO 20 mL, 2-fluoro-3- nitrotoluene (0.73 g, 4.7 mmol, 1.0 eq), potassium carbonate (1.95 g, 14.1 mmol, 3 eq) and 71A (1.00 g, 6.1 mmol, 1.3 eq) was dropped into the stirred mixture, then the mixture was stirred at 125°C for 60 hours. LC-MS showed the reaction was completed. This reaction was added water 50 mL and extracted with ethyl acetate (20 mL*3). The organic solution was dried over sodium sulfate, fdtered and concentrated. The residue was purified by column chromatography on silica gel to afford the compound 72A (0.8 g). Yield: 66.7%. [00416] Step 2\ To a 100 mL round-bottom flask was added methanol 20 mL, compound 72A (0.8 g, 3.1 mmol, 1.0 eq), Pd/C (0.2 g) was dropped into the stirred mixture at room temperature under H2 for overnight. LC-MS showed the reaction was completed. The solution was filtered and concentrated, the residue was purified by column chromatography on silica gel to afford the compound 72B (215 mg). Yield: 31.0%.

[00417] Step 3: To a 25 mL round-bottom flask was added 5 mL pyridine, compound 72B (176 mg, 0.76 mmol, 1.1 eq) and 4D (200 mg, 0.69 mmol, 1.0 eq), the reaction was stirred at room temperature for overnight. LC-MS showed the reaction was completed, the mixture was concentrated in vacuum and the residue was purified by column chromatography on silica gel to yield the title product 72 (136 mg, HPLC: 98.6%) as a white solid. Yield: 40.8%. Overall Yield: 8.4%. [M+l] ⁺ = 486.1. ¹ H NMR (400 MHz, CDCI ₃ ) δ 9.10 (br., 1H), 7.51 (d, 2H), 7.36 (d, 1H), 7.12 (t,lH), 6.90 (d, 1H), 3.29 (dd, 2H), 2.75 (s, 6H), 2.39 (d, 3H), 2.35-2.15 (m, 2H), 1.64 - 1.26 (m, 6H), 0.98 (d, 3H), 0.88 (dt, 3H).

Example 74: Synthesis of N2-(3-chloro-2-(4,4-diethylpiperidin-l-yl)phenyl) -N5,N5- dimethylthiophene-2,5-disulfonamide

[00418] Step 1. To a 500 ml three-neck flask bottom was added 200 ml toluene, 3-pentanone (62 g, 721 mmol, 1.0 eq), ethyl cyanoacetate (114 g, 1.01 mol, 1.4 eq), ammonium acetate (14.82 g, 192 mmol, 0.25 eq), acetic acid (51.89 g, 865 mmol, 1.2 eq). The flask was equipped with a Dean-Stark tube. The solution was refluxed under N2 overnight. GC showed the solution was nearly complete. The solution was cooled and poured into 1 L water. The water phase was extracted by 500ml methyl tert-butyl ether. The organic phase was washed with water (700 ml*2), brine (500 ml*2) dried over Na ₂ SO ₄ and concentrated under vacuum. Distillation under 0.2 torr to afford 60A (74 g) as a colorless liquid. Yield=62%.

[00419] Step 2'. To a 500 ml three-neck flask bottom was added 200 ml ethanol. Sodium (5.58 g, 242 mmol, 2.0 eq) was added with water bath. After the sodium had reacted, cyanoacetamide (20.38 g, 242 mmol, 2.0 eq) was added and the reaction mixture was heated to reflux for 15 min. The mixture was cooled and a solution of 60A (20 g, 121 mmol, 1.0 eq) in 20 ml ethanol was added as a thin stream. The reaction mixture, which was very thick, turned yellow-orange and was mildly exothermic. Stirring was continued overnight, and then the slurry was poured into 400 ml water, forming a homogeneous orange solution. Hydrochloric acid was added under ice bath to adjust pH to 2-3. The solution was stirred for lOmin under ice bath and the solid was collected by suction filtration, washed with cold water and dried under infrared. Another 54 g 60A was used to make compound 60B and total 60 g compound 60B was obtained as a white fluffy solid. Yield=61%.

[00420] Step 3 To a 500 ml three-neck flask was added 141 ml H2SO4, 60B (60 g, 274 mmol, 1.0 eq). The flask was equipped with a magnetic stir bar and a constant pressure dropping funnel. With stirring, 45.45 ml of water was added dropwise over a 45 min period. Sometimes solids emerged and then redissolved during this procedure. The solution was heated to reflux. After 1 h this moderated, and a clear yellow-brown solution with little foam was evident. Heating was discontinued, and after 10 min, 54.54 ml of water was added dropwise over a 35-min period to the still hot solution. Then heating was resumed, and the reaction mixture was brought to reflux overnight. The solution was cooled to 30°C and was poured into 200 ml water. The solution was filtered. The obtained solid was triturated 2 times with ether (350 ml*2). The ether phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 60C (40 g) as a white fluffy solid. Yield=78%.

[00421] Step 4. To a 3 L three-necked flask bottom was added urea (358 g, 5.97 mol, 29 eq), 60C (40 g, 207 mmol, 1.0 eq). The solution was heated to 160°C for 1 h under N2. Then heating was discontinued and 1 L water was added. After the solution was cooled to 30°C, it was extracted by ethyl acetate (300 ml*2). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 60D (14 g) as a white solid. Yield=39%.

[00422] Step 5: To a 1 L three-neck flask bottom was added 300 ml THF. LiAlHi (9.44 g, 248.52 mmol, 3.0 eq) was added at 0°C. 60D (14 g, 82.84 mmol, 1.0 eq) was added to the reaction mixture at 0-5°C. The solution was stirred at 25°C for 1 h and then refluxed for 3 h. The solution was cooled and 9.44 ml water, 18 ml 10% NaOH, 9.44 ml water was dropped to the solution under 5°C. The mixture was filtered and the solid was washed with DCM. The organic phase was dried over NaiSOi and concentrated under vacuum to afford compound 60E (11 g, GC=85%) as an orange oil. It was used to next step without purification. Yield=94%.

[00423] Step 6. To a 250 ml three-necked flask bottom was added 150 ml dioxane, 60E (11 g, 77.88 mmol, 1.0 eq), l-chloro-2-fluoro-3 -nitrobenzene (13.67 g, 77.88 mmol, 1.0 eq), K ₂ CO ₃ (32.24 g, 233.63 mmol, 3.0 eq). The reaction mixture was stirred at 100°C overnight. TLC showed the reaction was complete. The solution was cooled, poured into 500ml water and extracted by methyl tert-butyl ether (300 ml*2). The organic phase was dried over NaiSOi, concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether to afford compound 74A (15.5 g) as an orange solid. Yield=67.2%.

[00424] Step 7: To a 500 ml single-mouth flask bottom was added 250 ml MeOH, 74A (15.5 g, 52.36 mmol, 1.0 eq), 2 g Raney Ni. The reaction mixture was stirred at 15°C overnight under H2. TLC showed the reaction was complete. The solution was filtered. The organic phase was concentrated under vacuum and redissolved in 250 ml DCM. The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum to afford compound 74B (14.3 g) as a yellow liquid. It became solid after being placed overnight in the refrigerator. Yield=100%.

[00425] Step 8. To a 250 ml three-necked flask bottom was added 110 ml pyridine, 74B (11.1 g, 41.76 mol, 1.1 eq), 4D (11 g, 37.96 mmol, 1.0 eq). The reaction mixture was stirred for 4h. LC-MS showed the reaction was complete. The solution was concentrated under vacuum and purified by column chromatography on silica gel eluted with petroleum ether : ethyl acetate=5:l to afford compound 74 (11.9 g) as a light-yellow solid. Yield=60.4%. Overall yield=4.39%. [M+l] ⁺ = 520.1. 1H NMR (400 MHz, CDCI ₃ ) δ 9.06 (s, 1H), 7.61 (dd, 1H), 7.53 (d, 1H), 7.39 (d, 1H), 7.18 (t, 1H), 7.10 (dd, 1H), 3.63 (t, 2H), 2.75 (s, 6H), 2.15 (d, 2H), 1.59 (d, 2H), 1.52 (q, 2H), 1.41 (td, 2H), 1.32 (q, 2H), 0.85 (t, 3H), 0.80 (t, 3H).

Example 75: N5-[2-(6-azaspiro [2.5] octan-6-yl)-3-fluoro-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00426] Step 1. To a solution of 1, 2-difluoro-3 -nitro-benzene (500 mg, 3.14 mmol, 1 eq) and 6- azaspiro [2.5]octane (557 mg, 3.77 mmol, 1.2 eq, HC1) in ACN (5 mL) was added TEA (636 mg, 6.29 mmol, 875 μL, 2 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed 1, 2- difluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The solution was concentrated under vacuum. The obtained oil was added water (40 mL) and extracted by EtOAc (60 mL). The EtOAc phase was washed by saturated citric acid (30 mL). The water phase was extracted by EtOAc (30 mL). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum to give desired 6-(2-fluoro-6-nitro-phenyl)-6-azaspiro [2.5] octane (590 mg, crude) as orange oil. MS (ESI): mass calcd. For C13H15FN2O2 250.27, m/z found 251.0 [M+H] ⁺ .

[00427] Step 2: To a solution of 6-(2-fluoro-6-nitro-phenyl)-6-azaspiro [2.5] octane (590 mg, 2.36 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (0.5 mL) was added Fe (658 mg, 11.8 mmol, 5 eq) and NH4CI (1.26 g, 23.6 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed 6-(2-fluoro-6-nitro-phenyl)-6-azaspiro [2.5] octane was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was added H ₂ O (50 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 2-(6-azaspiro[2.5]octan-6-yl)-3-fluoro-aniline (410 mg, crude) as a black oil. MS (ESI): mass calcd. For C13H17FN2 220.29, m/z found 221.1 [M+H] ⁺ .

[00428] Step 3. To a solution of 2-(6-azaspiro[2.5]octan-6-yl)-3-fluoro-aniline (100 mg, 454 pmol, 1 eq) in Py (2 mL) was added 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (197 mg, 681 pmol, 1.5 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 2-(6-azaspiro [2.5] octan-6-yl)-3 -fluoro-aniline remained and desired compound was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition; column: Phenomenex Gemini-NX 80*40mm*3pm; mobile phase: [water (10mM NH ₄ HCO)-ACN]; B%: 45%-75%, 8mins) to give desired N5-[2-(6-azaspiro [2.5] octan-6-yl)-3 -fluoro-phenyl] -N2, N2-dimethyl-thiophene-2, 5- disulfonamide (67.2 mg, 141 pmol, 31.0% yield, 99.3% purity) as a light yellow solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 9.73 - 9.25 (m, 1 H) 7.71 (d, J= 4.03 Hz, 1 H) 7.66 (d, J= 4.03 Hz, 1 H) 7.32 - 7.21 (m, 2 H) 7.13 - 7.04 (m, 1 H) 2.66 (s, 9 H) 1.55 - 1.24 (m, 4 H) 0.30 (s, 4 H). HPLC: 99.28% (220 nm), 99.20% (215 nm), 99.51% (254 nm). MS (ESI): mass calcd. For C19H24FN3O4S3 473.60 m/z found 474.1 [M+H] ⁺ .

Example 76: N-[2-(6-azaspiro [2.5] octan-6-yl)-3-fluoro-phenyl]-5-pyrrolidin-l-ylsulfonyl- thiophene-2-sulfonamide

[00429] Step 1. To a solution of 2-(6-azaspiro [2.5]octan-6-yl)-3-fluoro-aniline (100 mg, 454 pmol, 1 eq) in Py (2 mL) was added 5-pyrrolidin-l-ylsulfonylthiophene-2-sulfonyl chloride (215 mg, 681 pmol, 1.5 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 2-(6-azaspiro [2.5] octan-6-yl)-3 -fluoro-aniline remained and desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition; column: Waters Xbridge BEH C18 100*25mm*5pm; mobile phase: [water (10 mM NEEHCO3)-ACN]; B%: 55%-85%, 10 mins) to give desired N-[2-(6- azaspiro[2.5]octan-6-yl)-3-fluoro-phenyl]-5-pyrrolidin-l-yls ulfonyl-thiophene-2-sulfonamide (67.2 mg, 134 pmol, 29.4% yield, 98.9% purity) as a light yellow solid. ¹ H NMR (400 MHz, DMSO-d6,) δ 9.50 (br s, 1 H) 7.70 (br s, 2 H) 7.34 - 7.18 (m, 2 H) 7.15 - 6.97 (m, 1 H) 3.14 (br s, 4 H) 2.66 (br s, 4 H) 1.66 (br s, 4 H) 1.41 (br s, 4 H) 0.30 (br s, 4 H). HPLC: 98.90% (220 nm), 98.83% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C21H26FN3O4S3 499.64 m/z found 500.2 [M+H] ⁺ .

Example 77: N-[2-(4, 4-dimethyl-l-piperidyl)-3-methyl-phenyl]-5-pyrrolidin-l-ylsu lfonyl- thiophene-2-sulfonamide

[00430] Step 1. To a solution of 5-bromothiophene-2-si/lfonyl chloride (10 g, 38.2 mmol, 1 eq) in DCM (100 mL) was added pyrrolidine (13.6 g, 191 mmol, 16.0 mL, 5 eq) at 0 °C. The mixture was stirred at 20 °C for 3 hours. LC-MS showed 5-bromothiophene-2-sulfonyl chloride was consumed completely and one main peak with desired mass was detected. The reaction mixture was partitioned between HC1 (IM, 40 mL) and DCM (150 mL). The organic phase was separated, washed with brine (80 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired l-[(5-bromo-2-thienyl)sulfonyl]pyrrolidine (12 g, crude) as a yellow oil. MS (ESI): mass calcd. For C ₈ Hi ₀ BrNO2S2 294.93, m/z found 295.9 [M+H] ⁺ .

[00431] Step 2'. To a solution of l-[(5-bromo-2-thienyl) sulfonyl] pyrrolidine (5 g, 16.9 mmol, 1 eq) and phenylmethanethiol (2.31 g, 18.6 mmol, 2.18 mL, 1.1 eq) and DIEA (4.36 g, 33.8 mmol, 5.88 mL, 2 eq) in Tol. (50 mL) was added Pd(dppf)C12 (309 mg, 422 pmol, 0.025 eq) and (5- diphenylphosphanyl-9, 9-dimethyl-xanthen-4-yl)-diphenyl-phosphane (977 mg, 1.69 mmol, 0.1 eq) under N2. The mixture was heat to 110 °C under N2 atmosphere and stirred for 12 hours. LC-MS showed l-[(5-bromo-2-thienyl) sulfonyl] pyrrolidine was consumed completely and one main peak with desired mass was detected. The mixture was cooled at 20 °C and added H ₂ O (200 mL), and extracted with EtOAc (300 mL). The combined organic layers were washed with brine (200 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Coli/mn, Eli/ent of 0-30% EtOAc/petroleum ether gradient @ 65 mL/min) to give desired l-[(5- benzylsulfanyl-2-thienyl)sulfonyl]pyrrolidine (3.8 g, 11.2 mmol, 66.3% yield) as a yellow solid. MS (ESI): mass calcd. For C15H17NO2S3 339.04, m/z found 340.0 [M+H+49] ⁺

[00432] Step 3 To a solution of l-[(5-benzylsulfanyl-2-thienyl) sulfonyl] pyrrolidine (2 g, 5.89 mmol, 1 eq) in AcOH (48 mL) and H ₂ O (12 mL) was added NCS (2.36 g, 17.7 mmol, 3 eq). The mixture was stirred at 20 °C for 4 hours. TLC showed l-[(5-benzylsulfanyl-2-thienyl) sulfonyl] pyrrolidine was consumed completely and one major new spot with larger polarity was detected. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (450 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-30% EtOAc/petrolewm ether gradient @ 40 mL/min) to give desired 5-pyrrolidin-l-ylsulfonylthiophene-2- sulfonyl chloride (1.6 g, 5.07 mmol, 86.0% yield) as a white solid.

[00433] Step 4. To a solution of 5-pyrrolidin-l-ylsulfonylthiophene-2-sulfonyl chloride (174 mg, 549 pmol, 1.2 eq) in Py. (2 mL) was added 2-(4, 4-dimethyl-l-piperidyl)-3-methyl-aniline (100 mg, 458 pmol, 1 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 2-(4, 4-dimethyl-l- piperidyl)-3-methyl-aniline was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WL,C (neutral condition; column: Waters Xbridge BEH C18 100*25mm*5i/m; mobile phase: [water (10mM NEEHCO3)-ACN]; B%: 65%-85%, lOmin) to give desired N-[2-(4,4- dimethyl-l-piperidyl)-3-methyl-phenyl]-5-pyrrolidin-l-ylsulf onyl-thiophene-2-sulfonamide (18.1 mg, 36.2 pmol, 7.90% yield, 99.6% pwrity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 0.98 (br s, 6 H) 1.30 - 1.50 (m, 4 H) 1.61 - 1.69 (m, 4 H) 2.30 (s, 3 H) 2.46 (br s, 2 H) 3.02 (br s, 2 H) 3.11 - 3.17 (m, 4 H) 6.97 - 7.12 (m, 3 H) 7.66 (d, J= 3.88 Hz, 1 H) 7.70 (d, J= 3.88 Hz, 1 H) 9.32 (s, 1 H). HPLC: 99.55% (220 nm), 99.26% (215 nm), 99.75% (254 nm). MS (ESI): mass calcd. For C22H31N3O4S3 497.15 m/z found 498.0 [M+H] ⁺ .

Example 78: N5-[2-(7-azaspiro [3.5] nonan-7-yl)-3-chloro-phenyl]-N2, N2-dimethyl-thiophene- 2, 5-disulfonamide.

[00434] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (452 mg, 2.58 mmol, 1 eq) and 7-azaspiro [3.5] nonane (500 mg, 3.09 mmol, 1.2 eq, HC1) in ACN (3 mL) was added TEA (521 mg, 5.15 mmol, 2 eq). The mixture was stirred at 80 °C for 2 hours. LC-MS showed l-chloro-2- fluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The solution was concentrated under vacuum. The obtained oil was added water (40 mL) and extracted by EtOAc (60 mL). The EtOAc phase was washed by saturated citric acid (30 mL). The water phase was extracted by EtOAc (30 mL). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum to give desired 7-(2-chloro-6-nitro-phenyl)-7-azaspiro [3.5] nonane (700 mg, crude) as a yellow solid. MS(ESI): mass calcd. For C14H17CIN2O2 280.10, m/z found 280.9 [M+H] ⁺ .

[00435] Step 2\ To a solution of 7-(2-chloro-6-nitro-phenyl)-7-azaspiro [3.5] nonane (700 mg, 2.49 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (0.5 mL) was added Fe (696 mg, 12.5 mmol, 5 eq) and NH ₄ CI (1.33 g, 24.9 mmol, 10 eq). The mixture was stirred at 80 °C for 2 hours. LC-MS showed 7- (2-chloro-6-nitro-phenyl)-7-azaspiro [3.5] nonane was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was added H ₂ O (50 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 2-(7-azaspiro[3.5]nonan-7-yl)-3-chloro-aniline (600 mg, crude) as a black oil. MS (ESI): mass calcd. For C14H19CIN2 250.12, m/z found 251.3 [M+H] ⁺ .

[00436] Step 3. To a solution of 2-(7-azaspiro [3.5] nonan-7-yl)-3 -chloro-aniline (100 mg, 399 pmol, 1 eq) in Py (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (173 mg, 598 pmol, 1.5 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 2-(7-azaspiro [3.5] nonan-7-yl)-3 -chloro-aniline was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (10 mM NTLHCO3)-ACN]; B%: 55%-80%, 8 mins) to give desired N5 - [2-(7-azaspiro [3.5 ]nonan-7-yl)-3 -chloro-phenyl] -N2,N2-dimethyl-thiophene-2, 5 - disulfonamide (60.7 mg, 119 pmol, 29.7% yield, 98.5% purity) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.54 (s, 1 H), 7.64 - 7.76 (m, 2 H), 7.25 - 7.31 (m, 2 H), 7.17 - 7.23 (m, 1 H), 3.06 - 3.26 (m, 2 H), 2.67 (s, 6 H) 2.21 - 2.40 (m, 2 H), 1.82 - 1.92 (m, 2 H), 1.72 - 1.80 (m, 4 H), 1.62 (br s, 4 H). HPLC: 98.49% (220 nm), 97.86% (215 nm), 99.30% (254 nm). MS (ESI): mass calcd. For C20H26CIN3O4S3 503.08 m/z found 504.1 [M+H] +.

Example 79: N2-(3-fluoro-2-(7-azaspiro [3.5] nonan-7-yl) phenyl)-N5, N5-dimethylthiophene-2, 5-disulfonamide

[00437] Step 1. To a solution of 1, 2-difluoro-3 -nitro-benzene (410 mg, 2.58 mmol, 1 eq) and 7- azaspiro [3.5] nonane (500 mg, 3.09 mmol, 1.2 eq, HC1) in ACN (3 mL) was added TEA (522 mg, 5.15 mmol, 2 eq). The mixture was stirred at 80 °C for 2 hours. LC-MS showed 2-difluoro-3 -nitrobenzene was consumed completely and desired mass was detected. The solution was concentrated under vacuum. The obtained oil was added H ₂ O (40 mL) and extracted by EtOAc (20 mL * 3). The EtOAc phase was washed by saturated citric acid (30 mL). The water phase was extracted by EtOAc (30 mL). The organic phase was dried over Na ₂ SO ₄ and concentrated under vacuum to give desired 7-(2-fluoro-6-nitrophenyl)-7- azaspiro [3.5] nonane (650 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C14H17FN2O2 265.13, m/z found 265.0 [M+H] ⁺ . [00438] Step 2\ To a solution of 7-(2-fluoro-6-nitro-phenyl)-7-azaspiro [3.5] nonane (650 mg, 2.46 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (0.5 mL) was added Fe (687 mg, 12.3 mmol, 5 eq) and NH ₄ CI (1.32 g, 24.6 mmol, 10 eq). The mixture was stirred at 80 °C for 2 hours. LC-MS showed 7- (2-fluoro-6-nitro-phenyl)-7-azaspiro [3.5] nonane was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was added H ₂ O (50 mL) and extracted with EtOAc (50 mL * 3). The combined organic layers were washed with brine 30 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 3-fluoro-2-(7- azaspiro [3.5] nonan-7-yl) aniline (550 mg, crude) as a black oil. MS (ESI): mass calcd. For C14H19FN2 234.15, m/z found 235.3 [M+H] ⁺ .

[00439] Step 3. To a solution of 2-(7-azaspiro [3.5] nonan-7-yl)-3 -fluoro-aniline (100 mg, 427 pmol, 1 eq) in Py (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (186 mg, 640 pmol, 1.5 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed Reactant 1 was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WY,C (neutral condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (10mM NH ₄ HCO ₃ )-ACN]; B%: 50%-80%, 8min) to give desired N2-(3-fluoro-2-(7- azaspiro [3.5] nonan-7-yl) phenyl)-N5, N5 -dimethylthiophene-2, 5 -disulfonamide (15 mg, 30.5 pmol, 7.14% yield, 99.02% purity) as a yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.53 (s, 1 H) 7.71 - 7.65 (m, 2 H) 7.32 - 7.21 (m, 2 H) 7.11 - 7.01 (m, 1 H) 2.67 (s, 6 H) 2.58 - 2.54 (m, 4 H) 1.91 - 1.82 (m, 2 H) 1.78 - 1.73 (m, 4 H) 1.61 (t, J= 5.13 Hz, 4 H). HPLC: 99.02% (220 nm), 98.74% (215 nm), 99.04% (254 nm). MS (ESI): mass calcd. For C20H26FN3O4S3487.11, m/z found 488.1 [M+H] ⁺ .

Example 80: N-[2-(6- azaspiro [2.5] octan-6-yl)-3-chloro-phenyl]-5-pyrrolidin-l-ylsulfonyl- thiophene-2-sulfonamide

[00440] Step 1. The mixture of 1 -chloro-2-fluoro-3 -nitro-benzene (200 mg, 1.14 mmol, 1 eq) and 6-azaspiro [2.5] octane (185 mg, 1.25 mmol, 1.1 eq, HC1) and K ₂ CO ₃ (472 mg, 3.42 mmol, 3 eq) in DMSO (2 mL) was heated to 120 °C and stirred for 12 hours under N2 atmosphere. LC-MS showed no l-chloro-2-fluoro-3 -nitro-benzene remained and desired compound was detected. The mixture was diluted with water (20 mL) and EtOAc (20 mL). The organic layer was separated and then washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give desired 6-(2-chloro-6-nitro-phenyl)-6- azaspiro [2.5] octane (200 mg, crude) as a white solid, which was used for the next step without further purification. MS (ESI): mass calcd. For C13H15CIN2O2 266.08, m/z found 266.9 [M+H] ⁺ .

[00441] Step 2 To a solution of 6-(2-chloro-6-nitro-phenyl)-6-azaspiro[2.5]octane (200 mg, 749 pmol, 1 eq) and NH4CI (401 mg, 7.50 mmol, 10 eq) in EtOH (3 mL) and H ₂ O (1 mL) was added Fe (209 mg, 3.75 mmol, 5 eq) at 80 °C under N2. The mixture was stirred at 80 °C for 36 hours. LC- MS showed 6-(2-chloro-6-nitro-phenyl)-6-azaspiro [2.5] octane was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water 10 mL and extracted with EtOAc (50 mL * 3). The combined organic layers were washed with brine 90 mL (30 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-20% EtOAc/petroleum ether gradient @ 30 mL/min) to give desired 2-(6- azaspiro [2.5] octan-6-yl)-3 -chloro-aniline (170 mg, 718 pmol, 95.8% yield) as a yellow oil. MS (ESI): mass calcd. For C13H17CIN2 236.11, m/z found 237.0 [M+H] ⁺ .

[00442] Step 3. To a solution of 5-pyrrolidin-l-ylsulfonylthiophene-2-sulfonyl chloride (140 mg, 444 pmol, 1.5 eq) in PYRIDINE (2 mL) was added 2-(6-azaspiro [2.5] octan-6-yl)-3 -chloro- aniline (70 mg, 296 pmol, 1 eq). The mixture was stirred at 15 °C for 12 hours. LC-MS showed 5- pyrrolidin-l-ylsulfonylthiophene-2-sulfonyl chloride was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition: column: Phenomenex Gemini-NX C18 75*30mm*3um; mobile phase: [water (10mM NH ₄ HHCO ₃ ]; B%: 35%-80%, 8 min) to give desired N-[2-(6-azaspiro[2.5]octan-6-yl)-3-chloro-phenyl]-5-pyrrolid in-l-ylsulfonyl- thiophene2-sulfonamide (11.7 mg, 31.0 pmol, 10.5% yield) as a white solid. ¹ H NMR (400MHz, DMSO-d6,) δ 9.53 (s, 1H), 7.77 - 7.70 (m, 2H), 7.34 - 7.26 (m, 2H), 7.24 - 7.17 (m, 1H), 3.27 (br s, 2H), 3.15 (br t, J= 6.6 Hz, 4H), 2.44 (br s, 2H), 1.88 - 1.74 (m, 2H), 1.71 - 1.62 (m, 4H), 1.07 (br s, 2H), 0.33 (br d, J= 8.8 Hz, 4H). HPLC: 98.22% (220 nm), 98.58% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C21H26CIN3O4S3515.08 m/z found 516.1 [M+H] ⁺ .

Example 81: N5-[3-chloro-2-[4-(trifluoro methyl)- 1-piperidyl] phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00443] Step 1. To a solution of 4-(trifluoromethyl) piperidine (480 mg, 3.13 mmol, 1.1 eq) and 1 -chloro-2-fluoro-3 -nitro-benzene (500 mg, 2.85 mmol, 1 eq) in dioxane (5 mL) was added K ₂ CO ₃ (787 mg, 5.70 mmol, 2 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 4- (trifluoromethyl) piperidine remained and desired compound was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 10/1) to give desired l-(2-chloro-6-nitro- phenyl)-4-(trifluoromethyl) piperidine (490 mg, 1.59 mmol, 55.7% yield) as an orange oil. MS (ESI): mass calcd. For C12H12CIF3N2O2 308.05, m/z found 309.2 [M+H] ⁺ .

[00444] Step 2 To a solution of l-(2-chloro-6-nitro-phenyl)-4-(trifluoromethyl) piperidine (490 mg, 1.59 mmol, 1 eq) in H ₂ O (5 mL) and EtOH (0.5 mL) was added Fe (443 mg, 7.94 mmol, 5 eq) and NH4CI (849 mg, 15.9 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed l-(2-chloro-6-nitro-phenyl)-4-(trifluoromethyl) piperidin was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was added H ₂ O (50 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine 30 (mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- TLC (SiC>2, petroleum ether/ EtOAc = 10/1) to give desired 3-chloro-2-[4-(trifluoromethyl)-l- piperidyl] aniline (400 mg, 1.44 mmol, 90.4% yield) as a yellow oil. MS (ESI): mass calcd. For C12H14CIN2F3 278.08, m/z found 279.3 [M+H] ⁺ .

[00445] Step 3. To a solution of 3 -chloro-2-[4-(trifluoromethyl)-l -piperidyl] aniline (100 mg, 359 pmol, 1 eq) in Py (2 mL) was added 5-(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (156 mg, 538 pmol, 1.5 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 3-chloro-2- [4-(trifluoromethyl)-l -piperidyl] aniline remained and desired compound was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-WLC (neutral condition; column: Waters Xbridge BEH C18 100*25mm*5um; mobile phase: [water (10mM NH ₄ HCO ₃ ); B%: 55%-75%, lOmin) to give desired N5-[3-chloro-2-[4- (trifluoromethyl)-l -piperidyl] phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (102 mg, 187 pmol, 52.1% yield, 97.6% purity) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.64 (br s, 1 H) 7.69 (s, 2 H) 7.41 - 7.52 (m, 1 H) 7.28 (br s, 2 H) 3.26 - 3.32 (m, 1 H) 3.13 - 3.22 (m, 1 H) 2.67 (br s, 6 H) 2.23 - 2.42 (m, 3 H) 1.76 (br s, 2 H) 1.69 (br s, 2 H). HPLC: 97.64% (220 nm), 97.48% (215 nm), 99.06% (254 nm). MS (ESI): mass calcd. For C18H21CIF3N3O4S3 531.03 m/z found 531.8 [M+H] ⁺ .

Example 82: N5-[2-(3, 3-dimethylpyrrolidin-l-yl)-3-methyl-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00446] Step 1. To a solution of 2-fluoro-l-methyl-3 -nitro-benzene (1 g, 6.45 mmol, 1 eq) and 3, 3-dimethylpyrrolidine (918 mg, 6.77 mmol, 1.05 eq, HC1) in DMSO (10 mL) was added CS ₂ CO ₃ (5.25 g, 16.1 mmol, 2.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 2- fluoro-l-methyl-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-20% Ethyl acetate/petroleum ether gradient @ 40 mL/min) to give desired 3, 3 -dimethyl- 1 -(2- methyl-6-nitro-phenyl) pyrrolidine (0.83 g, 3.54 mmol, 55.0% yield) as a yellow oil. MS (ESI): mass calcd. For C13H18N2O2 234.14, m/z found 235.1 [M+H] ⁺ .

[00447] Step 2'. To a solution of 3, 3 -dimethyl- l-(2-methyl-6-nitro-phenyl) pyrrolidine (500 mg, 2.13 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (1 mL) was added Fe (596 mg, 10.7 mmol, 5 eq) and NH4CI (1.14 g, 21.3 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed 3, 3 -dimethyl- l-(2-methyl-6-nitro-phenyl) pyrrolidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-15% Ethyl acetate/Petroleum ether gradient @ 30 mL/min) to give desired 2-(3, 3-dimethylpyrrolidin-l-yl)-3-methyl-aniline (220 mg, 1.08 mmol, 50.5% yield) as a yellow oil. MS (ESI): mass calcd. For C13H20N2204.16, m/z found 205.0 [M+H] ⁺ . [00448] Step 3: To a solution of 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (374 mg, 1.29 mmol, 1.2 eq) in pyridine (2 mL) was added 2-(3, 3-dimethylpyrrolidin-l-yl)-3-methyl-aniline (220 mg, 1.08 mmol, 1 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 5- (dimethylsulfamoyl) thiophene-2-sulfonyl chloride was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by pre p-HPLC (neutral condition: column: Phenomenex Gemini-NX 80*40mm*3um; mobile phase: [water (10mM NELHCO ₃ -ACN]; B%: 45%-75%, 8min) to give desired N5-[2-(3, 3-dimethylpyrrohdin-l-yl)-3-methyl-phenyl]-N2, N2-dimethyl-thiophene- 2, 5 -disulfonamide (118 mg, 258 pmol, 24.0% yield) as a yellow solid. ¹ H NMR (400MHz, DMSO- d ₆ ) 5 9.37 (br s, 1H), 7.65 (br d, J= 17.0 Hz, 2H), 7.19 - 6.97 (m, 3H), 3.09 (br s, 2H), 2.66 (br s, 6H), 2.62 (br s, 2H), 2.22 (br s, 3H), 1.71 (br s, 2H), 1.14 (br s, 6H). HPLC: 95.64% (220 nm), 96.44% (215 nm), 97.76% (254 nm). MS (ESI): mass calcd. For C19H27N3O4S3 457.12 m/z found 458.0 [M+H] ⁺ .

Example 83: 5-[4-chloro-3-(4,4-dimethyl-l-piperidyl) indazol-l-yl] sulfonyl-N,N-dimethyl- thiophene-2-sulfonamide

[00449] Step 1. To a solution of 3, 3 -dimethylazetidine (823 mg, 6.77 mmol, 1.05 eq, HC1) in DMSO (10 mL) was added 2-fluoro-l-methyl-3 -nitro-benzene (1 g, 6.45 mmol, 1 eq) and CS ₂ CO ₃ (5.25 g, 16.1 mmol, 2.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 2- fluoro-l-methyl-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-15% Ethyl acetate/petroleum ether gradient @ 40 mL/min) to give desired 3, 3 -dimethyl- 1 -(2- methyl-6-nitro-phenyl)azetidine (1.3 g, 5.90 mmol, 91.6% yield) as a yellow solid. MS (ESI): mass calcd. For C12H16N2O2 220.12, m/z found 221.0 [M+H] ⁺ .

[00450] Step 2: To a solution of 3, 3 -dimethyl- l-(2-methyl-6-nitro-phenyl) azetidine (500 mg, 2.27 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (1 mL) was added Fe (634 mg, 11.4 mmol, 5 eq) and NH4CI (1.21 g, 22.7 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed 3, 3 -dimethyl- l-(2-methyl-6-nitro-phenyl) azetidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaElash® Silica Elash Column, Eluent of 0-15% Ethyl acetate/Petroleum ether gradient @ 30 mL/min) to give desired 2-(3, 3-dimethylazetidin-l-yl)-3-methyl-aniline (320 mg, 1.68 mmol, 74.1% yield) as a yellow oil. MS (ESI): mass calcd. For C12H18N2 190.15, m/z found 191.0 [M+H] ⁺ .

[00451] Step 3. To a solution of 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (365 mg, 1.26 mmol, 1.2 eq) in pyridine (2 mL) was added 2-(3,3-dimethylazetidin-l-yl)-3-methyl-aniline (200 mg, 1.05 mmol, 1 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 2-(3, 3- dimethylazetidin-l-yl)-3-methyl-aniline was consumed completely and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition: column: Phenomenex Luna C18 75*30mm*3um; mobile phase: [water (10mM NH4HCOs)-ACN]; B%: 50%-80%, 8min ) to give desired N5-[2-(3,3- dimethylazetidin-l-yl)-3-methyl-phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (55.3 mg, 123 pmol, 11.7% yield, 98.6% purity) as a gray solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.74 (s, 1H), 7.69 (d, J= 3.9 Hz, 1H), 7.53 (d, J= 3.9 Hz, 1H), 6.91 (d, J= 7.2 Hz, 1H), 6.45 (t, J= 7.6 Hz, 1H), 6.24 (d, J= 7.6 Hz, 1H), 3.89 (s, 4H), 2.71 (s, 6H), 2.21 (s, 3H), 1.24 (s, 6H). HPLC: 98.59% (220 nm), 98.60% (215 nm), 99.56% (254 nm). MS (ESI): mass calcd. For C18H25N3O4S3 443.10 m/z found 443.9 [M+H] ⁺ .

Example 84: N-(2-(4, 4-dimethylpiperidin-l-yl)-3-methylphenyl)-5-(isopropylsulfon yl) thiophene-2-sulfonamide

[00452] Step 1. To a solution of 2-(4, 4-dimethylpiperidin-l-yl)-3 -methylaniline (0.5 g, 2.29 mmol, 1 eq) in DCM (5 mL) was added 5-bromothiophene-2-sulfonyl chloride (659 mg, 2.52 mmol, 1.1 eq), TEA (463 mg, 4.58 mmol, 637 μL, 2 eq). The mixture was stirred at 20 °C for 12 hours.

LC-MS showed was consumed completely and one main peak with desired mass was detected. Then it was separated between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0~6% Ethyl acetate/petroleum ether gradient @ 50 mL/min) to give desired 5-bromo-N-(2-(4, 4-dimethylpiperidin-l-yl)-3 -methylphenyl) thiophene-2- sulfonamide (0.6 g, 1.35 mmol, 59.09% yield) as a yellow solid. MS (ESI): mass calcd. For Ci8H ₂₃ BrN2O2S2 442.04, m/z found 443.0 [M+H] ⁺ .

[00453] Step 2 To a solution of 5-bromo-N-(2-(4, 4-dimethylpiperidin-l-yl)-3 -methylphenyl) thiophene-2-sulfonamide (200 mg, 451 pmol, 1 eq) and 2-(isopropyldisulfanyl)propane (142 mg, 947 pmol, 151 μL, 2.1 eq) in THF (2 mL) was added dropwise w-BuLi (2.5 M, 235 μL, 1.3 eq) at -70 °C. The resulting mixture was stirred at -70 °C for 0.5 hour. LC-MS showed 5-bromo-N-(2-(4, 4- dimethylpiperidin-l-yl)-3 -methylphenyl) thiophene-2-sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was separated between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep-TLC (SiCL, petroleum ether/Ethyl acetate = 5/1) to give desired N-(2-(4, 4- dimethylpiperidin-l-yl)-3-methylphenyl)-5-(isopropylthio) thiophene-2-sulfonamide (90 mg, 205 pmol, 45.5% yield) as a yellow oil. MS (ESI): mass calcd. For Ci8H2 ₃ BrN2O2S2 442.04, m/z found 443.0 [M+H] ⁺ .

[00454] Step 3. To a solution of N-(2-(4, 4-dimethylpiperidin-l-yl)-3-methylphenyl)-5- (isopropylthio) thiophene-2-sulfonamide (90 mg, 205 pmol, 1 eq) in DCM (1 mL) was added m- CPBA (125 mg, 616 pmol, 85% purity, 3 eq) at 0 °C. The mixture was stirred at 20 °C for 12 hours. LC-MS showed N-(2-(4, 4-dimethylpiperidin-l-yl)-3-methylphenyl)-5-(isopropylthio) thiophene-2- sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was partitioned between 5 mL of sat. Na ₂ SO ₃ and 10 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of sat. Na ₂ SO ₃ , ImL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep- HPLC (neutral condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (10mM NEEHCO ₃ )-ACN]; B%: 50%-80%,10min) to give desired N-(2-(4, 4- dimethylpiperidin-l-yl)-3-methylphenyl)-5-(isopropylsulfonyl )thiophene-2-sulfonamide (30 mg, 80 pmol, 30.2% yield, 97.29% purity) as a yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.40 (br s, 1H), 7.78 (br d, J =3.3 Hz, 1H), 7.67 (br d, J =3.5 Hz, 1H), 7.10 - 6.96 (m, 3H), 3.58 (td, J= 6.4, 13.1 Hz, 1H), 3.01 (br s, 2H), 2.29 (s, 3H), 1.48 - 1.28 (m, 4H), 1.18 (br d, J= 6.6 Hz, 6H), 0.97 (br s, 6H). HPLC: 97.29% (220 nm), 97.74% (215 nm), 99.81% (254 nm). MS (ESI): mass calcd. For C21H30N2O4S3 470.14 m/z found 471.2 [M+H] ⁺ .

Example 85: N2-(3-chloro-2-(4, 4-difluoropiperidin-l-yl) phenyl)-N5, N5-dimethylthiophene-2, 5-disulfonamide

[00455] Step 1. To a solution of 4, 4-difluoropiperidine (500 mg, 3.17 mmol, 1 eq, HC1) in DMSO (5 mL) was added l-chloro-2-fluoro-3 -nitro-benzene (613 mg, 3.49 mmol, 1.1 eq) and CS ₂ CO ₃ (2.58 g, 7.93 mmol, 2.5 eq). The mixture was stirred at 100 °C for 12 hours. TLC (petroleum ether/EtOAc=5/l, Rf = 0.64) indicated no 4, 4-difluoropiperidine remained, and one major new spot with lower polarity was detected. The reaction mixture was diluted with H ₂ O (20 mL) and extracted with (EtOAc). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-20% EtOAc/petroleum ether gradient @ 40 mL/min) to give desired l-(2-chloro-6-nitrophenyl)-4, 4-difluoropiperidine (230 mg, 831 pmol, 26.2% yield) as a yellow oil.

[00456] Step 2 To a solution of l-(2-chloro-6-nitro-phenyl)-4, 4-difluoro-piperidine (230 mg, 831 pmol, 1 eq) in EtOH (3 mL) and H ₂ O (0.6 mL) was added Fe (232 mg, 4.16 mmol, 5 eq) and NH4CI (445 mg, 8.31 mmol, 10 eq). The mixture was stirred at 80 °C for 3 hours. LC-MS showed 1- (2-chloro-6-nitro-phenyl)-4, 4-difluoro-piperidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (10 mL) and extracted with (EtOAc). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, PE/EtOAc = 3/1) to give desired 3-chloro-2-(4, 4- difluoropiperidin-l-yl) aniline (140 mg, 568 pmol, 68.3% yield) as a yellow oil. MS (ESI): mass calcd. For C11H13CIF2N2 246.07, m/z found 247.1 [M+H] ⁺ .

[00457] Step 3. To a solution of 3-chloro-2-(4, 4-difluoro-l -piperidyl) aniline (140 mg, 568 pmol, 1 eq) in PYRIDINE (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (197 mg, 681 pmol, 1.2 eq). The mixture was stirred at 15 °C for 3 hours. LC-MS showed 3-chloro- 2-(4, 4-difluoro-l -piperidyl) aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition: column: Phenomenex Gemini -NX Cl 8 75*30mm*3um; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 35%-55%, 8min ) to give desired N2-(3-chloro-2-(4, 4-difluoropiperidin-l-yl) phenyl)-N5, N5 -dimethylthiophene-2, 5- disulfonamide (70 mg, 140.00 pmol, 24.7% yield, 99.87% purity) as a yellow oil. ¹ H NMR (400MHz, DMSO-c/z) δ 8.52 (s, 1H), 7.58 (dd, J= 1.4, 8.2 Hz, 1H), 7.51 (d, J= 3.9 Hz, 1H), 7.38 (d, J= 3.9 Hz, 1H), 7.21 - 7.16 (m, 1H), 7.11 - 7.08 (m, 1H), 3.69 (br t, J= 11.4 Hz, 2H), 2.76 (s, 6H), 2.44 (br dd, J= 3.2, 12.0 Hz, 2H), 2.21 - 2.10 (m, 2H), 2.10 - 1.91 (m, 2H). HPLC: 99.87% (220 nm), 99.82% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C17H20CIF2N3O4S3 499.03 m/z found 500.1 [M+H] ⁺ .

Example 86: N2, N2-dimethyl-N5-(3-methyl-2-(2-azaspiro [3.3] heptan-2-yl) phenyl) thiophene- 2, 5-disulfonamide

[00458] To a solution of 2-fluoro-l-methy 1-3 -nitro-benzene (276 mg, 1.78 mmol, 1 eq) and 2- azaspiro[3.3]heptane (250 mg, 1.87 mmol, 1.05 eq, HC1) in DMSO (4 mL) was added CS ₂ CO ₃ (1.45 g, 4.45 mmol, 2.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 2-fluoro-l- methyl-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (90 mL). The combined organic layers were washed with brine (40 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 4/1) to give desired 2-(2-methyl-6-nitro-phenyl)-2- azaspiro[3.3]heptane (200 mg, 861 pmol, 48.32% yield) as a yellow oil. MS (ESI): mass calcd. For C13H16N2O2 232.12, m/z found 233.0 [M+H] ⁺ .

[00459] Step 2'. To a solution of 2-(2-methyl-6-nitro-phenyl)-2-azaspiro[3.3]heptanes (200 mg, 861 pmol, 1 eq) in MeOH (10 mL) was added Pd/C (10 mg, 50% purity) and H2 (861 pmol). The mixture was stirred at 20 °C for 1 hour. LC-MS showed 2-(2-methyl-6-nitro-phenyl)-2-azaspiro [3.3] heptanes was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and washed with EtOAc (90 mL), then the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0-15% EtOAc/petroleum ether gradient @ 30 mL/min) to give desired 2-(2-azaspiro [3.3] heptan-2-yl)-3-methyl-aniline (150 mg, 741 pmol, 86.1% yield) as a yellow oil. MS (ESI): mass calcd. For C13H18N2 202.15, m/z found 203.0 [M+H] ⁺ .

[00460] Step 3. To a solution of 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (236 mg, 816 pmol, 1.1 eq) in pyridine (2 mL) was added 2-(2-azaspiro[3.3]heptan-2-yl)-3-methyl-aniline (150 mg, 741 pmol, 1 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 2-(2- azaspiro[3.3]heptan-2-yl)-3-methyl-aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-W ,C (neutral condition: column: Phenomenex Gemini- NX C18 75*30mm*3um; mobile phase: [water(10mM NtLHCCLl-ACN]; B%: 40%-70%, lOmin), but impurity (MS: 492) accounted for about 50%, then the residue was second purified by prep- HPLC (FA condition: column: Phenomenex Luna C18 75*30mm*3um; mobile phase: [water(0.2%FA)-ACN]; B%: 20%-70%,8min) to give desired N2, N2-dimethyl-N5-(3-methyl-2-(2- azaspiro [3.3] heptan-2-yl) phenyl) thiophene-2, 5-disulfonamide (31.3 mg, 66.9 pmol, 9.02% yield, 99.06% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.73 (br s, 1H), 7.66 (d, J =

3.9 Hz, 1H), 7.50 (d, J= 3.9 Hz, 1H), 6.88 (d, J = 7.1 Hz, 1H), 6.44 (t, J = 7.6 Hz, 1H), 6.25 (d, J =

6.9 Hz, 1H), 4.10 (s, 4H), 2.69 (s, 6H), 2.19 (s, 3H), 2.14 - 2.06 (m, 4H), 1.83 - 1.73 (m, 2H). HPLC: 99.06% (220 nm), 98.98% (215 nm), 99.48% (254 nm). MS (ESI): mass calcd. For C19H25N3O4S3 455.10 m/z found 456.1 [M+H] ⁺ .

Example 87: N5-[2-(3, 3a, 4, 5, 6, 6a-hexahydro-lH-cyclopenta[c]pyrrol-2-yl)-3-methyl-phenyl]-

N2, N2-dimethyl-thiophene-2, 5-disulfonamide

[00461] Step 1. To a solution of 2-fluoro-l-methyl-3 -nitro-benzene (500 mg, 3.22 mmol, 1 eq) and 1, 2, 3, 3a, 4, 5, 6, 6a-octahydrocyclopenta[c]pyrrole (571 mg, 3.87 mmol, 1.2 eq, HC1) in dioxane (4 mL) was added K ₂ CO ₃ (891 mg, 6.45 mmol, 2 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 2-fluoro-l-methyl-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCE, petroleum ether/EtOAc = 1/0 to 2/1) to give desired 2-(2-methyl-6-nitro- phenyl)-3, 3a, 4, 5, 6, 6a-hexahydro-lH-cyclopenta[c]pyrrole (631 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C14H18N2O2 246.14, m/z found 247.3 [M+H] ⁺ .

[00462] Step 2.' A mixture of 2-(2-methyl-6-nitro-phenyl)-3, 3a, 4, 5, 6, 6a-hexahydro-lH- cyclopenta[c]pyrrole (400 mg, 1.62 mmol, 1 eq), Pd/C (50 mg, 10% purity, 1 eq) in MeOH (10 mL) was degassed and purged with H2 (15 PSI) for 3 times, and then the mixture was stirred at 20 °C for 12 hours under H2 atmosphere. LC-MS showed 2-(2-methyl-6-nitro-phenyl)-3, 3a, 4, 5, 6, 6a- hexahydro-lH-cyclopenta[c]pyrrole was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCE, petroleum ether/EtOAc = 5/1) to give desired 2-(3, 3a, 4, 5, 6, 6a-hexahydro-lH-cyclopenta[c]pyrrol-2-yl)-3-methyl-aniline (180 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C14H20N2 216.16, m/z found 217.1 [M+H] ⁺ .

[00463] Step 3: To a solution of 2-(3, 3a, 4, 5, 6, 6a-hexahydro-lH-cyclopenta[c]pyrrol-2-yl)-3- methyl-aniline (160 mg, 740 pmol, 1 eq) in Py (1 mL) was added 5-(dimethylsulfamoyl)thiophene-2- sulfonyl chloride (322 mg, 1.11 mmol, 1.5 eq). The mixture was stirred at 20 °C for 2 hours. LC- MS showed 2-(3, 3a, 4, 5, 6, 6a-hexahydro-lH-cyclopenta[c]pyrrol-2-yl)-3-methyl-aniline was consumed completely and desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by pre p-HPLC (neutral condition; column: Phenomenex Gemini- NX 80*40mm*3um; mobile phase: [water (10mM NELHCO3)-ACN]; B%: 45%-75%, 8min) to give desired N5-[2-(3, 3a, 4, 5, 6, 6a-hexahydro-lH-cyclopenta[c]pyrrol-2-yl)-3-methyl-phenyl]-N 2, N2- dimethyl-thiophene-2, 5 -disulfonamide (142 mg, 301 pmol, 40.7% yield, 100% purity) as a yellow solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 9.44 (s, 1 H), 7.75 - 7.70 (m, 1 H), 7.69 - 7.66 (m, 1 H), 7.21 - 7.10 (m, 2 H), 7.08 - 7.01 (m, 1 H), 3.19 (br t, J= 7.82 Hz, 2 H), 2.75 - 2.72 (m, 1 H), 2.71 (s, 6 H), 2.66 (s, 1 H), 2.62 (br d, J= 8.38 Hz, 2 H), 2.29 (s, 3 H), 1.82 (br dd, J= 12.19, 5.32 Hz, 2 H), 1.78- 1.69 (m, 1 H), 1.60 (dt, J= 11.79, 6.05 Hz, 1 H), 1.51 - 1.41 (m, 2 H). HPLC: 100.00% (220 nm), 100.00% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C20H27N3O4S3 469.12 m/z found 470.2 [M+H] ⁺ .

Example 88: 5-[[2-(4, 4-dimethyl-l-piperidyl)-3-methyl-phenyl] sulfamoyl] -N, N-dimethyl- thiophene-2-carboxamide

[00464] Step 1. To a solution of 5-bromo-N-[2-(4,4-dimethyl-l-piperidyl)-3-methyl-phenyl] thiophene-2-sulfonamide (0.3 g, 677 pmol, 1 eq), N-methylmethanamine (110 mg, 1.35 mmol, 124 μL, 2 eq, HC1) and TEA (205 mg, 2.03 mmol, 283 μL, 3 eq) in MeOH (10 mL) was added Pd(dppf)C12 (49.5 mg, 67.7 pmol, 0.1 eq) under CO atmosphere. The suspension was degassed and purged with CO for 3 times. The mixture was stirred under CO (19.0 mg, 677 pmol, 15.2 μL) (50 Psi) at 70 °C for 12 hours. LC-MS showed 5-bromo-N-[2-(4, 4-dimethyl-l-piperidyl)-3 -methylphenyl] thiophene-2-sulfonamide was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition; column: Waters Xbridge BEH C18 100*25mm*5um;mobile phase: [water(10mM NH ₄ HCO ₃ )-ACN];B%: 50%-80%,10min) to give desired 5-[[2-(4,4-dimethyl-l-piperidyl)-3-methyl-phenyl] sulfamoyl]-N, N-dimethyl-thiophene-2-carboxamide (20.5 mg, 45.8 pmol, 6.76% yield, 97.2% purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.14 (s, 1 H) 7.52 (d, J= 4.03 Hz, 1 H) 7.47 (d, J= 3.91 Hz, 1 H) 7.19 (d, J = 7.82 Hz, 1 H) 7.07 (t, J = 7.76 Hz, 1 H) 6.97 (d, J= 7.46 Hz, 1 H) 3.21 - 2.93 (m, 8 H) 2.46 - 2.35 (m, 2 H) 2.32 (s, 3 H) 1.46 (br d, J= 2.08 Hz, 2 H) 1.37 (br s, 2 H) 0.99 (br d, J= 8.44 Hz, 6 H). HPLC: 97.24% (220 nm), 97.40% (215 nm), 99.57% (254 nm). MS (ESI): mass calcd. For C21H29N3O3S2 435.17 m/z found 436.10 [M+H] ⁺ .

Example 89: N-[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] -5-isopro pylsulfonyl- thiophene- 2-sulfonamide [00465] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (1 g, 5.70 mmol, 1 eq) and 4,4-dimethylpiperidine (938 mg, 6.27 mmol, 1.1 eq, HC1) in dioxane (10 mL) was added K ₂ CO ₃ (2.36 g, 17.1 mmol, 3 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 1- chloro-2-fluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc=l/0 to 3/1) to give desired l-(2-chloro-6-nitro-phenyl)-4, 4-dimethyl-piperidine (1.5 g, crude) as ayellow solid. MS (ESI): mass calcd. For C13H17CIN2O2 268.10, m/z found 269.3 [M+H] ⁺ .

[00466] Step 2 To a solution of l-(2-chloro-6-nitro-phenyl)-4, 4-dimethyl-piperidine (1.5 g, 5.58 mmol, 1 eq) and NH4CI (2.99 g, 55.8 mmol, 10 eq) in H ₂ O (2 mL) and EtOH (20 mL) was added Fe (1.56 g, 27.9 mmol, 5 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed l-(2- chloro-6-nitro-phenyl)-4, 4-dimethyl-piperidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (40 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (90 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 1/0 to 2/1) to give desired 3-chloro-2-(4, 4-dimethyl-l -piperidyl) aniline (1.05 g, 4.40 mmol, 78.8% yield) as ayellow oil. MS (ESI): mass calcd. For C13H19CIN2 238.12, m/z found 239.3 [M+H] ⁺ .

[00467] Step 3. To a solution of 3-chloro-2-(4, 4-dimethyl-l -piperidyl) aniline (400 mg, 1.68 mmol, 1 eq) and 5-bromothiophene-2-sulfonyl chloride (876 mg, 3.35 mmol, 2 eq) in DCM (4 mL) was added TEA (339 mg, 3.35 mmol, 466 μL, 2 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 3-chloro-2-(4, 4-dimethyl-l -piperidyl) aniline was consumed completely and one main peak with desired mass was detected. Then it was separated between water (2 mL) and EtOAc (4 mL). The organic phase was separated, washed with brine (3 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0~6% EtOAc/petroleum ether gradient @ 50 mL/min) to give desired 5-bromo-N-[3-chloro-2-(4, 4- dimethyl-1 -piperidyl) phenyl] thiophene-2-sulfonamide (680 mg, 1.47 mmol, 87.5% yield) as a yellow solid. MS (ESI): mass calcd. For Ci7H2oBrClN202S2 461.98, m/z found 465.2 [M+H] ⁺ .

[00468] Step 4. To a solution of 5-bromo-N-[3-chloro-2-(4,4-dimethyl-l- piperidyl)phenyl]thiophene-2-sulfonamide (0.5 g, 1.08 mmol, 1 eq) and 1 ,2-diisopropyldisulfane (340 mg, 2.26 mmol, 361 μL, 2.1 eq) in THF (5 mL) was added dropwise n-BuLi (2.5 M, 561 μL, 1.3 eq) at -70 °C. The resulting mixture was stirred at -70 °C for 0.5 hour. LC-MS showed 5- bromo-N-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] thiophene-2-sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was separated between water (2 mL) and EtOAc (4 mL). The organic phase was separated, washed with brine (3 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep-TLC (SiO ₂ , petroleum ether/EtOAc = 5/1) to give desired N-[3-chloro-2-(4, 4- dimethyl-1 -piperidyl) phenyl]-5-isopropylsulfanyl-thiophene-2-sulfonamide (0.22 g, 479 pmol, 44.5% yield) as a yellow oil. MS (ESI): mass calcd. For C20H27CIN2O2S3 458.09, m/z found 459.1 [M+H] ⁺ .

[00469] Step 5: To a solution of N-[3-chloro-2-(4, 4-dimethyl-l-piperidyl)phenyl]-5- isopropylsulfanyl-thiophene-2-sulfonamide (220 mg, 479 pmol, 1 eq) in DCM (3 mL) was added m- CPBA (292 mg, 1.44 mmol, 85% purity, 3 eq) at 0 °C. The mixture was stirred at 20 °C for 12 hours. LC-MS showed N-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl]-5-isopropylsulfanyl- thiophene-2-sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was partitioned between sat. Na ₂ SO ₃ (5 mL) and EtOAc (10 mL). The organic phase was separated, washed with sat. Na ₂ SO ₃ (5 mL), brine (1 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep- HPLC (neutral condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (10mM NH ₄ HCO ₃ )-ACN]; B%: 50%-80%, 10 mins) to give desired N-[3-chloro-2-(4,4- dimethyl-l-piperidyl)phenyl]-5-isopropylsulfonyl-thiophene-2 -sulfonamide (47.8 mg, 92.7 pmol, 19.4% yield, 95.3% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ -,) δ 9.57 (s, 1H), 7.79 (d, J= 4.0 Hz, 1H), 7.69 (d, J= 4.0 Hz, 1H), 7.30 - 7.24 (m, 2H), 7.21 - 7.14 (m, 1H), 3.59 (quin, J= 6.8 Hz, 1H), 3.33 - 3.19 (m, 2H), 2.41 - 2.27 (m, 2H), 1.57 - 1.41 (m, 2H), 1.36 - 1.23 (m, 2H), 1.18 (d, J= 6.8 Hz, 6H), 0.96 (s, 6H). HPLC: 95.25% (220 nm), 96.01% (215 nm), 98.89% (254 nm). MS (ESI): mass calcd. For C20H27CIN2O4S3 490.08 m/z found 491.1 [M+H] ⁺ .

Example 90: N5-[2-(3-cyclobutylpyrrolidin-l-yl)-3-methyl-phenyl]-N2, N2-dimethyl-thiophene- 2, 5-disulfonamide

[00470] Step 1. To a solution of tert-butyl 0 -oxopyrrolidine- 1 -carboxylate (10 g, 54.0 mmol, 1 eq) in THF (100 mL) was added t-BuOK (14.5 g, 130 mmol, 2.4 eq) at 0 °C. The mixture was stirred at 0 °C for 0.5 hour. Then bromo-(4-bromobutyl)-triphenyl-X5-phosphane (31.0 g, 64.8 mmol, 1.2 eq) in THF (100 mL) was added dropwise to the mixture at 0 °C. Then the mixture was stirred at 20 °C for 12 hours. LC-MS showed tert-butyl 3 -oxopyrrolidine-1 -carboxylate was consumed completely and desired mass was detected. The reaction mixture was added to water (300 mL), the aqueous phase was extracted with EtOAc (600 mL). The organic layer were washed with brine (200 mL), dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 100/1 to 2/1) to give desired tert-butyl 3 -cyclobutylidenepyrrolidine-1 -carboxylate (1.25 g, 5.60 mmol, 10.4% yield) as ayellow oil. MS (ESI): mass calcd. For C13H21NO2 223.16, m/z found 168.1 [M-56] ⁺ .

[00471] Step 2'. A mixture of tert-butyl 3 -cyclobutylidenepyrrolidine-1 -carboxylate (1.25 g, 5.60 mmol, 1 eq), Pd/C (200 mg, 10% purity) in MeOH (30 mL) was degassed and purged with H2 for 3 times, and then the mixture was stirred at 20 °C for 12 hours under H2(15PSI) atmosphere. LC-MS showed tert-butyl 3 -cyclobutylidenepyrrolidine-1 -carboxylate was consumed completely and desired mass was detected. The reaction mixture was fdtered and the filtrate was concentrated under reduced pressure to give desired tert-butyl 3 -cyclobutylpyrrolidine-1 -carboxylate (1.1 g, crude) as ayellow oil. MS (ESI): mass calcd. For C13H23NO2 225.17, m/z found 170.1 [M-56] ⁺ .

[00472] Step 3. The mixture of tert-butyl 3 -cyclobutylpyrrolidine-1 -carboxylate (1.1 g, 4.88 mmol, 1 eq) in HCl/MeOH (10 mL) was stirred at 20 °C for 1 hour. LC-MS showed tert-butyl 3- cyclobutylpyrrolidine-1 -carboxylate was consumed completely and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give desired 3-cyclobutylpyrrolidine (750 mg, crude, HC1) as an orange oil. MS (ESI): mass calcd. For CsHisN 125.12, m/z found 126.3 [M+H] ⁺ .

[00473] Step 4. To a solution of 2-fluoro-l-methyl-3 -nitro-benzene (100 mg, 645 pmol, 1 eq) and 3-cyclobutylpyrrolidine (156 mg, 967 pmol, 1.5 eq, HC1) in dioxane (2 mL) was added K ₂ CO ₃ (267 mg, 1.93 mmol, 3 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 2- fluoro-l-methyl-3 -nitro-benzene desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCE, petroleum ether/EtOAc = 10/1) to give desired 3 -cyclobutyl- l-(2-methyl-6-nitro- phenyl) pyrrolidine (150 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C15H20N2O2 260.15, m/z found 261.1 [M+H] ⁺ .

[00474] Step 5: A mixture of 3 -cyclobutyl- l-(2-methyl-6-nitro-phenyl) pyrrolidine (180 mg, 691 pmol, 1 eq), Pd/C (50 mg, 320 pmol, 10% purity) in MeOH (5 mL) was degassed and purged with EEQ 5 PSI) for 3 times, and then the mixture was stirred at 20 °C for 1 hour under H2 atmosphere. LC-MS showed 3 -cyclobutyl- l-(2-methyl-6-nitro-phenyl) pyrrolidine was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give desired 2-(3-cyclobutylpyrrolidin-l-yl)-3-methyl-aniline (140 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C15H22N2 230.18, m/z found 231.1 [M+H] ⁺ .

[00475] Step 6 To a solution of 2-(3-cyclobutylpyrrolidin-l-yl)-3-methyl-aniline (90 mg, 391 pmol, 1 eq) in Py (1 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (170 mg, 586 pmol, 1.5 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed 2-(3- cyclobutylpyrrolidin-l-yl)-3-methyl-aniline was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCE, petroleum ether/EtOAc = 2/1) to give desired N5-[2-(3-cyclobutylpyrrohdin-l-yl)-3-methyl-phenyl]-N2, N2-dimethyl-thiophene-2, 5- disulfonamide (28.3 mg, 57.9 pmol, 20.0% yield, 99.0% purity) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 9.49 (br s, 1 H) 7.65 (br s, 2 H) 7.19 (br s, 1 H) 7.12 - 6.93 (m, 2 H) 2.97 (br d, J = 1.59 Hz, 1 H) 2.92 - 2.78 (m, 2 H) 2.65 (br s, 7 H) 2.32 (br s, 2 H) 2.19 (br s, 3 H) 1.95 (br s, 3 H) 1.81 (br s, 2 H) 1.72 - 1.59 (m, 2 H) 1.54 (br d, J= 1.59 Hz, 1 H). HPLC: 98.97% (220 nm), 98.85% (215 nm), 99.94% (254 nm). MS (ESI): mass calcd. For C21H29N3O4S3 483.13 m/z found 484.2 [M+H] ⁺ .

Example 91: 5-(azetidin-l-ylsulfonyl)-N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene-2-sulfonamide

[00476] Step 1. To a solution of 5-bromothiophene-2-sulfonyl chloride (1 g, 3.82 mmol, 1 eq) in DCM (10 mL) was added TEA (967 mg, 9.55 mmol, 1.33 mL, 2.5 eq) and azetidine (429 mg, 4.58 mmol, 507 μL, 1.2 eq, HC1). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 5- bromothiophene-2-sulfonyl chloride was consumed completely and desired mass was detected. The reaction mixture was quenched by water 30 mL, and then extracted with DCM 60 mL (20 mL * 3). The combined organic layers were washed with brine 20 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 1- [(5 -bromo-2-thienyl)sulfonyl] azetidine (1 g, crude) as an orange oil was used into the next step without further purification. MS (ESI): mass calcd. For C ₇ H8BrNO ₂ S2 280.92, m/z found 281.8 [M+H] ⁺ .

[00477] Step 2 A mixture of l-[(5-bromo-2-thienyl) sulfonyl] azetidine (1 g, 3.54 mmol, 1 eq), phenylmethanethiol (484 mg, 3.89 mmol, 456 μL, 1.1 eq), DIEA (915 mg, 7.08 mmol, 1.23 mL, 2 eq), Pd(dppf)C12 (64.8 mg, 88.5 pmol, 0.025 eq) and (5-diphenylphosphanyl-9, 9-dimethyl-xanthen- 4-yl)-diphenyl-phosphane (205 mg, 354 pmol, 0.1 eq) in Tol. (10 mL) was degassed and purged with N ₂ for 3 times, and then the mixture was stirred at 110 °C for 12 hours under N ₂ atmosphere. LC-MS showed l-[(5-bromo-2-thienyl) sulfonyl] azetidine was consumed completely and desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove Tol.. The mixture was cooled at 20 °C and added H ₂ O (100 mL), and extracted with EtOAc 150 mL (50 mL * 3). The combined organic layers were washed with brine 100 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/Ethyl acetate=l/O to 1/1) to give desired l-[(5- benzylsulfanyl-2-thienyl) sulfonyl] azetidine (1 g, crude) as a black oil. MS (ESI): mass calcd. For C14H15NO2S3325.03, m/z found 326.2 [M+H] ⁺ .

[00478] Step 3 To a solution of l-[(5-benzylsulfanyl-2-thienyl) sulfonyl] azetidine (1 g, 3.07 mmol, 1 eq) in AcOH (10 mL) and H ₂ O (2.5 mL) was added NCS (1.23 g, 9.22 mmol, 3 eq). The mixture was stirred at 20 °C for 4 hours. TLC (petroleum ether/Ethyl acetate =3/1) showed l-[(5- benzylsulfanyl-2-thienyl) sulfonyl] azetidine was consumed completely and one major new spot with larger polarity was detected. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCE, petroleum ether/Ethyl acetate=l/O to 0/1) to give desired 3-chloro-2-(4, 4-dimethyl-l -piperidyl) aniline (1 g, crude) as a white solid. MS (ESI): mass calcd. For C7ILCINO4S3 300.93, m/z found 302.1 [M+H] ⁺ .

[00479] Step 4. To a solution of 3-chloro-2-(4, 4-dimethyl-l -piperidyl) aniline (200 mg, 838 pmol, 1 eq) in Py (2 mL) was added 5-(azetidin-l-ylsulfonyl) thiophene-2-sulfonyl chloride (379 mg, 1.26 mmol, 1.5 eq). The mixture was stirred at 20 °C for 12 hours. LC-MS showed 3-chloro-2-(4, 4- dimethyl-1 -piperidyl) aniline was consumed completely and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/Ethyl acetate = 2/1) to give desired 5-(azetidin-l-ylsulfonyl)-N-(3- chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene-2-sulfonamide (24.8 mg, 48.7 pmol, 5.82% yield, 99.1% purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 9.58 (s, 1H), 7.79 (d, J= 2.8 Hz, 2H), 7.33 - 7.28 (m, 2H), 7.23 (d, J= 8.1 Hz, 1H), 3.74 (t, J= 7.6 Hz, 4H), 2.54 - 2.53 (m, 6H), 2.35 (br d, J= 1.7 Hz, 2H), 2.11 - 1.98 (m, 2H), 1.53 (br s, 2H), 1.33 (br s, 2H), 0.98 (br s, 6H), HPLC: 99.06% (220 nm), 99.25% (215 nm), 99.04% (254 nm). MS (ESI): mass calcd. For C20H26CIN3O4S3 503.08 m/z found 504.1 [M+H] ⁺ .

Example 92: N5-[3-chloro-2-(2-methyl-l-piperidyl) phenyl] -N2, N2-dimethyl- thiophene-2, 5- disulfonamide

[00480] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (1 g, 5.70 mmol, 1 eq) and 2- methylpiperidine (927 mg, 6.84 mmol, 1.11 mL, 1.2 eq, HC1) in dioxane (10 mL) was added K ₂ CO ₃ (1.97 g, 14.2 mmol, 2.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 1- chloro-2-fluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 1/0 to 2/1) to give desired l-(2-chloro-6-nitro-phenyl)-2-methyl-piperidine (522 mg, 2.05 mmol, 36.0% yield) as a yellow oil. MS (ESI): mass calcd. For C12H15CIN2O2254.08, m/z found 255.3 [M+l] ⁺ .

[00481] Step 2. To a solution of l-(2-chloro-6-nitro-phenyl)-2-methyl-piperidine (500 mg, 1.96 mmol, 1 eq) in EtOH (10 mL) and H ₂ O (1 mL) was added Fe (548 mg, 9.82 mmol, 5 eq) and NH4CI (1.05 g, 19.6 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed l-(2- chloro-6-nitro-phenyl)-2-methyl-piperidine was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The reaction mixture was added to water (30 mL), the aqueous phase was extracted with EtOAc (90 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 1/0 to 2/1) to give desired 3 -chloro-2-(2-methyl-l -piperidyl) aniline (380 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C12H17CIN2 224.11, m/z found 225.3 [M+l] ⁺ .

[00482] Step 3: To a solution of 3 -chloro-2-(2-methyl-l -piperidyl) aniline (150 mg, 667 pmol, 1 eq) in Py (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (290 mg, 1.00 mmol, 1.5 eq). The mixture was stirred at 15 °C for 12 hours. LC-MS showed 3-chloro-2-(2- methyl-1 -piperidyl) aniline was consumed completely and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 2/1) to give desired N5-[3-chloro-2-(2-methyl-l- piperidyl) phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (89.1 mg, 186 pmol, 27.8% yield, 99.7% purity) as a yellow solid. ¹ H NMR (400 MHz, DMSO-r/ ₆ ) δ 9.31 (s, 1 H) 7.83 (d, J = 4.03 Hz, 1 H) 7.70 (d, J= 4.03 Hz, 1 H) 7.63 - 7.54 (m, 1 H) 7.38 - 7.28 (m, 1 H) 7.24 (s, 1 H) 3.47 (br d, J = 6.36 Hz, 1 H) 3.14 (br d, J= 3.42 Hz, 1 H) 2.65 (s, 6 H) 1.95 (br d, J= 11.00 Hz, 1 H) 1.78 (br dd, J = 10.39, 1.34 Hz, 1 H) 1.68 (br d, J= 9.78 Hz, 1 H) 1.54 (br s, 2 H) 1.38 (br d, J= 9.54 Hz, 2 H) 0.52 (d, J= 6.36 Hz, 3 H). HPLC: 99.71% (220 nm), 99.71% (215 nm), 98.91% (254 nm). MS (ESI): mass calcd. For C18H24CIN3O4S3 477.06 m/z found 477.9 [M+H] ⁺ .

Example 93: N-[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] -5-cyclopen tylsulfonyl- thiophene-2-sulfonamide [00483] Step 1. To a solution of compound 5-bromo-N-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] thiophene-2-sulfonamide (0.2 g, 431 pmol, 1 eq), cyclopentanethiol (57.3 mg, 561 pmol, 60 μL, 1.3 eq), BINAP (268 mg, 431 pmol, 1 eq) and CS ₂ CO ₃ (351 mg, 1.08 mmol, 2.5 eq) in Tol. (2 mL) was added Pd2 (dba)s (197 mg, 216 pmol, 0.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed compound 5-bromo-N-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] thiophene-2-sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was separated between 5 mL of water and 8 mL of EtOAc. The organic phase was separated, washed with 5 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 100/1 to 50/1) to give desired N-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl]-5-cyclopentylsulfanyl-thiophene-2-sulfonamide (0.2 g, crude) as a yellow oil. MS (ESI): mass calcd. For C22H29CIN2O2S3 484.11 m/z found 485.1 [M+H] ⁺

[00484] Step 2: To a solution of N-[3-chloro-2-(4,4-dimethyl-l-piperidyl)phenyl]-5- cyclopentylsulfanyl-thiophene-2-sulfonamide (0.2 g, 412 pmol, 1 eq) in DCM (3 mL) was added m- CPBA (251 mg, 1.24 mmol, 85% purity, 3 eq )at 0 °C. The mixture was stirred at 20 °C for 12 hours. LC-MS showed N-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl]-5-cyclopentylsulfanyl-thiophene- 2-sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was partitioned between 5 mL of sat. Na ₂ SO ₃ and 10 mL of EtOAc. The organic phase was separated, washed with 5 mL of sat. Na ₂ SO ₃ , ImL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep- HPLC (column: Waters Xbridge BEH C18 100*30mm*10um;mobile phase: [water(10mM NH ₄ HCO ₃ )-ACN];B%: %-%,10min) to give desired N-[3-chloro-2-(4, 4-dimethyl-l- piperidyl)phenyl]-5-cyclopentylsulfonyl-thiophene-2-sulfonam ide (34.6 mg, 64.7 pmol, 15.7% yield, 96.7% purity) was obtained as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.58 (s, 1H), 7.83 (d, J= 4.0 Hz, 1H), 7.69 (d, J= 4.0 Hz, 1H), 7.31 - 7.24 (m, 2H), 7.22 - 7.14 (m, 1H), 3.94 (quin, J= 7.8 Hz, 1H), 3.27 (br s, 2H), 2.33 (br s, 2H), 1.84 (quind, J= 6.9, 14.2 Hz, 4H), 1.56 (td, J= 3.6, 6.8 Hz, 4H), 1.50 (br s, 2H), 1.31 (br s, 2H), 0.97 (br s, 6H). HPLC: 96.68% (220 nm), 97.05% (215 nm), 98.15% (254 nm). MS (ESI): mass calcd. For C22H29CIN2O4S3 516.10 m/z found 517.2 [M+H] ⁺ . Example 94: N2, N2-dimethyl-N5-[3-methyl-2-(3-phenylpyrrolidin-l-yl) phenyl] thiophene-2, 5- disulfonamide

[00485] Step 1. To a solution of 2-fluoro-l-methyl-3 -nitro-benzene (400 mg, 2.58 mmol, 1 eq) and 3-phenylpyrrolidine (569 mg, 3.87 mmol, 1.5 eq) in dioxane (6 mL) was added K ₂ CO ₃ (713 mg, 5.16 mmol, 2 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 2-fluoro-l- methyl-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCh, petroleum ether/EtOAc = 1/0 to 2/1) to give desired l-(2-methyl-6-nitro-phenyl)-3-phenyl-pyrrolidine (495 mg, 1.75 mmol, 68.0% yield) as a yellow oil. MS (ESI): mass calcd. For C17H18N2O2 282.14, m/z found 283.1[M+H] ⁺ .

[00486] Step 2: A mixture of l-(2-methyl-6-nitro-phenyl)-3-phenyl-pyrrolidine (495 mg, 1.75 mmol, 1 eq), Pd/C (50 mg, 320 pmol, 10% purity) in MeOH (5 mL) was degassed and purged with H2 (15 PSI) for 3 times, and then the mixture was stirred at 20 °C for 1 hour under H2 atmosphere. LC-MS showed l-(2-methyl-6-nitro-phenyl)-3-phenyl-pyrrolidine was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 5/1) to give desired 3-methyl-2-(3-phenylpyrrolidin-l-yl) aniline (398 mg, 1.58 mmol, 90.0% yield) as a yellow oil. MS (ESI): mass calcd. For C17H20N2252.16, m/z found 253.2 [M+H] ⁺ .

[00487] Step 3. To a solution of 3-methyl-2-(3-phenylpyrrolidin-l-yl) aniline (120 mg, 476 pmol, 1 eq) in Py (1 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (207 mg, 713 pmol, 1.5 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed 3-methyl-2-(3- phenylpyrrolidin-l-yl) aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 2/1) to give desired N2, N2-dimethyl-N5-[3-methyl-2-(3-phenylpyrrolidin-l-yl) phenyl] thiophene-2, 5 -disulfonamide (17.8 mg, 33.9 pmol, 16.4% yield, 96.2% purity) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.69 (br s, 1 H) 7.62 (br d, J= 2.69 Hz, 2 H) 7.35 (br d, J= 3.18 Hz, 5 H) 7.24 (br d, J= 6.85 Hz, 2 H) 7.09 (br d, J= 17.60 Hz, 2 H) 3.63 - 3.51 (m, 1 H) 3.21 - 3.10 (m, 2 H) 3.07 - 2.95 (m, 2 H) 2.63 (br d, J= 7.34 Hz, 1 H) 2.58 (s, 6 H) 2.28 (br s, 4 H) 2.19 - 2.10 (m, 1 H). HPLC: 99.21% (220 nm), 96.67% (215 nm), 100.0% (254 nm). MS (ESI): mass calcd. For C23H27N3O4S3 505.12 m/z found 506.2 [M+H] ⁺ .

Example 95: 5-(azetidin-l-ylsulfonyl)-N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene-2-sulfonamide

[00488] Step 1. To a solution of (2R, 6S)-2, 6-dimethylmorpholine (722 mg, 6.27 mmol, 1.1 eq) in DMSO (5 mL) was added CS ₂ CO ₃ (4.64 g, 14.2 mmol, 2.5 eq) and 1 -chloro-2-fluoro-3 -nitrobenzene (1 g, 5.70 mmol, 1 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed (2R, 6S)-2, 6-dimethylmorpholine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water 50 mL and extracted with EtOAc (300 mL). The combined organic layers were washed with brine (90 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0-15% Ethyl acetate/petroleum ether gradient @ 60 mL/min) to give desired (2S, 6R)-4-(2- chloro-6-nitro-phenyl)-2, 6-dimethyl-morpholine (600 mg, 2.22 mmol, 38.9% yield) as a yellow solid. MS (ESI): mass calcd. For C12H15CIN2O3 270.08, m/z found 217.2 [M+H] ⁺ .

[00489] Step 2: To a solution of (2S, 6R)-4-(2-chloro-6-nitro-phenyl)-2, 6-dimethyl-morpholine (300 mg, 1.11 mmol, 1 eq) in EtOH (3 mL) and H ₂ O (0.6 mL) was added Fe (619 mg, 11.1 mmol, 10 eq) andNH ₄ C1 (593 mg, 11.1 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed (2S, 6R)-4-(2-chloro-6-nitro-phenyl)-2, 6-dimethyl-morpholine was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with water 10 mL and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-25% Ethyl acetate/Petroleum ether gradient @ 30 mL/min) to give desired 3-chloro-2-[(2S, 6R)-2, 6-dimethylmorpholin-4-yl] aniline (180 mg, 748 pmol, 67.5% yield) as a pale yellow solid. MS (ESI): mass calcd. For C12H17CIN2O 240.10, m/z found 241.2 [M+H] ⁺ . [00490] Step 3: To a solution of 3-chloro-2-[(2S, 6R)-2, 6-dimethylmorpholin-4-yl] aniline (180 mg, 748 pmol, 1 eq) in pyridine (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (260 mg, 898 pmol, 1.2 eq). The mixture was stirred at 20 °C for 3 hours. LC-MS showed 3-chloro-2-[(2S, 6R)-2, 6-dimethylmorpholin-4-yl] aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition: column: Phenomenex Gemini-NX C18 75*30mm*3um; mobile phase: [water (10mM NH ₄ HCO ₃ ; B%: 45%-65%, 8 min) to give desired N2-(3-chloro-2-((2S, 6R)-2, 6-dimethylmorpholino)phenyl)-N5, N5- dimethylthiophene-2, 5-disulfonamide (194 mg, 384 pmol, 51.4% yield, 97.8% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.63 (br s, 1H), 7.70 - 7.66 (m, 1H), 7.64 (br s, 1H), 7.48 (br s, 1H), 7.30 (br s, 2H), 3.76 - 3.60 (m, 2H), 2.94 (br t, J=10.3 Hz, 2H), 2.66 (s, 6H), 2.12 - 1.92 (m, 2H), 0.99 (br s, 6H). HPLC: 97.84% (220 nm), 97.71% (215 nm), 98.29% (254 nm). MS (ESI): mass calcd. For C18H24CIN3O5S3 493.06 m/z found 494.1 [M+H] ⁺ .

Example 96: N2-(2-((3S, 4S)-3, 4-dimethylpyrrolidin-l-yl)-3-methylphenyl)-N5, N5- dimethylthiophene-2, 5-disulfonamide

[00491] Step 1. To a solution of 2-fluoro-l-methyl-3 -nitro- benzene (500 mg, 3.22 mmol, 1 eq) in DMSO (3 mL) was added CS ₂ CO ₃ (2.63 g, 8.06 mmol, 2.5 eq) and 3,4-dimethylpyrrolidine (481 mg, 3.55 mmol, 1.1 eq, HC1). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 2- fluoro-l-methyl-3 -nitro- benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL * 3). The combined organic layers were washed with brine (20 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-15% EtOAc/petroleum ether gradient @ 30 mL/min) to give desired 3, 4-dimethyl-l-(2-methyl-6- nitrophenyl) pyrrolidine (400 mg, 1.71 mmol, 52.97% yield) as a yellow oil. MS (ESI): mass calcd. For C13H18N2O2 234.14, m/z found 235.2 [M+H] ⁺ .

[00492] Step 2.' To a solution of 3, 4-dimethyl-l-(2-methyl-6-nitro-phenyl) pyrrolidine (400.00 mg, 1.71 mmol, 1 eq) in MeOH (20 mL) was added Pd/C (10%, 50 mg) under N2 atmosphere. The suspension was degassed and purged with H2 for 3 times. The mixture was stirred under H2 (15 Psi or atm.) at 15°C for 1 hour. LC-MS showed 3, 4-dimethyl-l-(2-methyl-6-nitro-phenyl) pyrrolidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and washed 3 times with EtOAc and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-20% Ethyl acetate/Petroleum ether gradient @ 35 mL/min) to give desired 2-(3, 4-dimethylpyrrolidin-l-yl)-3 -methylaniline (280 mg, 1.37 mmol, 80.3% yield) as a pale yellow oil. MS (ESI): mass calcd. For C13H20N2 204.16, m/z found 205.2 [M+H] ⁺ .

[00493] Step 3. To a solution of 2-(3, 4-dimethylpyrrolidin-l-yl)-3-methyl-aniline (280 mg, 1.37 mmol, 1 eq) in pyridine (3 mL) was added 5 -(dimethylsulfamoyl) thioph ene-2-sulfonyl chloride (437 mg, 1.51 mmol, 1.1 eq). The mixture was stirred at 15 °C for 3 hours. LC-MS showed 2-(3, 4- dimethylpyrrolidin-l-yl)-3-methyl-aniline was consumed completely and two main peaks with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove solvent to give a residue (420 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C19H27N3O4S3 457.12, m/z found 458.3 [M+H] ⁺ .

[00494] Step 4. N5-[2-(3,4-dimethylpyrrolidin-l-yl) -3-methyl-phenyl]-N2, N2-dimethyl- thiophene-2, 5 -disulfonamide (420 mg, 918 pmol, 1 eq) was purified by prep-WY,C ( HC1 condition, column: Phenomen ex luna Cl 8 80*40mm*3 um; mobile phase: [water(0.04%HCl)-ACN];B%: 58%- 73%,7min) to give desired compound which was diluted with 10 mL of water and basified with half saturated solution of NaHCCE to pH 8, and then it was extracted with EtOAc (10 mL), the organic layer was separated and washed with 10 mL of brine, dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give desired N2-(2-((3S, 4S)-3, 4- dimethylpyrrolidin-l-yl)-3-methylphenyl)-N5, N5 -dimethylthiophene-2, 5 -disulfonamide (26.5 mg, 55.12 pmol, 6.01% yield, 95.19% purity) as a pale red solid. ¹ H NMR (400MHZ, DMSO-d ₆ ,) δ 9.55 (br s, 1H), 7.64 (br d, J= 8.3 Hz, 2H), 7.19 (br s, 1H), 7.05 (br d, J= 19.5 Hz, 2H), 3.42 (br s, 2H), 2.91 (br s, 2H), 2.66 (br s, 6H), 2.20 (br s, 3H), 1.89 (br s, 2H), 1.00 (br s, 6H). HPLC: 95.19% (220 nm), 94.81% (215 nm), 98.83% (254 nm). MS (ESI): mass calcd. For C19H27N3O4S3 457.12 m/z found 458.0 [M+H] ⁺ .

Example 97: N2-(2-((3S, 4S)-3, 4-dimethylpyrrolidin-l-yl)-3-methylphenyl)-N5, N5- dimethylthiophene-2, 5-disulfonamide

[00495] Step 1. N5-[2-(3, 4-dimethylpyrrolidin-l-yl)-3-methyl-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide 5 (420 mg, 918 pmol, 1 eq) was purified by prep-HPLC ( HC1 condition : column: Phenomenex luna Cl 8 80*40mm*3 um; mobile phase: [water(0.04%HCl)- ACN]; B%: 58%-73%, 7min) to give desired N2-(2-((3R, 4S)-3, 4-dimethylpyrrolidin-l-yl)-3- methylphenyl)-N5, N5 -dimethylthiophene-2, 5-disulfonamide which had few impurity, then purified by second prep-HPLC (neutral condition: column: Waters Xbridge BEH Cl 8 100*25mm*5um;mobile phase: [water(10mM NH ₄ HCO ₃ )-ACN];B%: 50%-75%,10min) to give desired N2-(2-((3R, 4S)-3, 4-dimethylpyrrolidin-l-yl)-3-methylphenyl)-N5, N5-dimethylthiophene- 2, 5-disulfonamide (61.2 mg, 133 pmol, 14.5% yield, 99.6% purity) as a white solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 9.29 (s, 1H), 7.65 (dd, J= 3.9, 16.9 Hz, 2H), 7.15 - 6.99 (m, 3H), 3.10 (br t, J= 7.3 Hz, 2H), 2.66 (s, 6H), 2.59 (br dd, J= 4.9, 7.9 Hz, 2H), 2.41 - 2.31 (m, 2H), 2.21 (s, 3H), 0.97 (br d, J= 6.3 Hz, 6H). HPLC: 99.62% (220 nm), 99.52% (215 nm), 99.55% (254 nm). MS (ESI): mass calcd. For C19H27N3O4S3 457.12 m/z found 458.0 [M+H] ⁺ .

Example 98: N2-(3-chloro-2-(4-methoxy-4-methylpiperidin-l-yl) phenyl)-N5, N5- dimethylthiophene-2, 5-disulfonamide [00496] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (233 mg, 1.33 mmol, 1.1 eq) and 4-methoxy-4-methyl-piperidine; hydrochloride (200 mg, 1.21 mmol, 1 eq) in DCM (5 mL) was added TEA (366 mg, 3.62 mmol, 504 μL, 3 eq). The mixture was stirred at 40 °C for 12 hours. LCMS showed l-chloro-2-fluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The crude was added H ₂ O (10 mL), and extracted with EtOAc 45 mL (15 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/Ethyl acetate=l/O to 80/1) to give desired l-(2-chloro-6-nitro-phenyl)-4-methoxy-4-methyl-piperidine (225 mg, 790 pmol, 65.5% yield) as a white solid. MS (ESI): mass calcd. For C13H17CIN2O3 284.09, m/z found 284.9 [M+H] ⁺ .

[00497] Step 2: To a solution of l-(2-chloro-6-nitro-phenyl)-4-methoxy-4-methyl-piperidine (225 mg, 790 pmol, 1 eq) in EtOH (3 mL) and H ₂ O (0.3 mL) was added Fe (353 mg, 6.32 mmol, 8 eq) and NH4CI (634 mg, 11.9 mmol, 15 eq). The mixture was stirred at 80 °C for 12 hours. LCMS showed l-(2-chloro-6-nitro-phenyl)-4-methoxy-4-methyl-piperidine was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude oil. The crude was added H ₂ O (30 mL), and extracted with EtOAc 60 mL (20 mL * 3). The combined organic layers were washed with brine 30 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/Ethyl acetate=4/l to 3/1) to give desired 3-chloro-2- (4-methoxy-4-methyl-l -piperidyl) aniline (137 mg, 538 pmol, 68.1% yield) as a white solid. MS (ESI): mass calcd. For C13H19CIN2O 254.12, m/z found 255.0 [M+H] ⁺ .

[00498] Step 3. To a solution of 3 -chloro-2-(4-methoxy-4-methyl-l -piperidyl) aniline (130 mg, 510 pmol, 1 eq) in Py (2 mL) was added 5-(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (177.45 mg, 612.35 pmol, 1.2 eq). The mixture was stirred at 15 °C for 12 hours. LCMS showed 3- chloro-2-(4-methoxy-4-methyl-l -piperidyl) aniline was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a crude oil. The residue was purified by prep-TLC (SiCL, petroleum ether/Ethyl acetate = 3:1). The purified product was dissolved in 20 mL of water, and then the solution was lyophilized to give desired N2-(3-chloro-2-(4- methoxy-4-methylpiperidin-l-yl) phenyl)-N5, N5 -dimethylthiophene-2, 5 -disulfonamide (38.9 mg, 74.2 pmol, 14.5% yield, 96.9 % purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.51 (br s, 1H), 7.70 (br s, 2H), 7.46 - 7.32 (m, 1H), 7.31 - 7.25 (m, 1H), 7.25 - 6.94 (m, 1H), 3.47 - 3.37 (m, 2H), 3.35 - 3.27 (m, 1H), 3.13 (br s, 3H), 2.67 (s, 6H), 2.15 - 1.99 (m, 1H), 1.81 - 1.51 (m, 4H), 1.16 (s, 3H). HPLC: 96.89% (220 nm), 95.54% (215 nm), 99.68% (254 nm). MS (ESI): mass calcd. For C19H26CIN3O5S3507.07 m/z found 508.1 [M+H] ⁺ .

Example 99: N5-[3-chloro-2-(3, 3-difluoro-l-piperidyl) phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00499] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (2.34 g, 13.3 mmol, 1.05 eq) and 3, 3 -difluoropiperidine; hydrochloride (2 g, 12.7 mmol, 1 eq) in dioxane (40 mL) was added K ₂ CO ₃ (5.26 g, 38.1 mmol, 3 eq). The mixture was stirred at 100 °C for 60 hours. LCMS showed 1- chloro-2-fluoro-3 -nitro-benzene was consumed and desired mass was detected. The crude was added H ₂ O (60 mL), and extracted with EtOAc (150 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc=l/0 to 80/1) to give desired a l-(2-chloro- 6-nitro-phenyl)-3, 3 -difluoro-piperidine (0.9 g, crude) as a brown oil. MS (ESI): mass calcd. For C11H11CIF2N2O2 276.05, m/z found 276.8 [M+H] ⁺ .

[00500] Step 2: To a solution of l-(2-chloro-6-nitro-phenyl)-3, 3 -difluoro-piperidine (0.9 g, 3.25 mmol, 1 eq) in EtOH (10 mL) and H ₂ O (1 mL) was added Fe (1.45 g, 26.0 mmol, 8 eq) and NH4CI (2.61 g, 48.8 mmol, 15 eq). The mixture was stirred at 80 °C for 12 hours. LCMS showed l-(2- chloro-6-nitro-phenyl)-3, 3 -difluoro-piperidine was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude oil. The crude was added H ₂ O (40 mL), and extracted with EtOAc (90 mL). The combined organic layers were washed with brine 50 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 4/1 to 3/1) to give desired a 3-chloro-2-(3, 3 -difluoro- 1 -piperidyl) aniline (600 mg, 2.43 mmol, 74.8% yield) as a brown oil. MS (ESI): mass calcd. For C11H13CIF2N2 246.07, m/z found 247.0 [M+H] ⁺ .

[00501] Step 3. To a solution of 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (200 mg, 690 pmol, 1 eq) in Py (2 mL) was added 3-chloro-2-(3, 3 -difluoro- 1 -piperidyl) aniline (170 mg, 690 pmol, 1 eq). The mixture was stirred at 15 °C for 12 hours. LCMS showed 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a crude oil. The residue was purified by prep-TLC (SiCh, Petroleum ether: EtOAc = 3/1). The purified product was dissolved in water (20 mL), and then the solution was lyophilized to give desired N5-[3-chloro-2-(3, 3 -difluoro- 1 -piperidyl) phenyl] - N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (54.7 mg, 109 pmol, 15.9% yield, 100.0% purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.45 - 9.41 (m, 1H), 7.71 (s, 2H), 7.36 (dd, J= 1.5, 7.9 Hz, 1H), 7.27 - 7.21 (m, 1H), 3.47 - 3.36 (m, 2H), 3.32 - 3.21 (m, 1H), 3.12 - 2.92 (m, 1H), 2.68 (s, 6H), 2.17 - 1.96 (m, 2H), 1.89 - 1.72 (m, 2H). HPLC: 100.0% (220 nm), 99.64% (215 nm), 100.0% (254 nm). MS (ESI): mass calcd. For C17H20CIF2N3O4S3 499.03, m/z found 500.2 [M+H] ⁺ .

Example 100: N5-[3-chloro-2-(2, 6-dimethylmorpholin-4-yl) phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00502] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (480 mg, 2.74 mmol, 1.05 eq) and 2, 6-dimethylmorpholine (300 mg, 2.60 mmol, 321 μL, 1 eq) in dioxane (10 mL) was added K ₂ CO ₃ (1.08 g, 7.81 mmol, 3 eq). The mixture was stirred at 100 °C for 12 hours. LCMS showed 1- chloro-2-fluoro-3 -nitro-benzene was consumed and desired mass was detected. The crude was added H ₂ O (20 mL), and extracted with EtOAc (45 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 1/0 to 80/1) to give 4-(2-chloro-6-nitro- phenyl)-2, 6-dimethyl-morpholine (300 mg, 1.11 mmol, 42.5% yield) as a brown oil. MS (ESI): mass calcd. For C12H15CIN2O3270.08, m/z found 270.9 [M+H] ⁺ .

[00503] Step 2: To a solution of 4-(2-chloro-6-nitro-phenyl)-2, 6-dimethyl-morpholine (300 mg, 1.11 mmol, 1 eq) in EtOH (3 mL) and H ₂ O (0.3 mL) was added Fe (495 mg, 8.87 mmol, 8 eq) and NH4CI (889 mg, 16.6 mmol, 15 eq). The mixture was stirred at 80 °C for 12 hours. LCMS showed 4-(2-chloro-6-nitro-phenyl)-2, 6-dimethyl-morpholine was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude oil. The crude was added H ₂ O (10 mL), and extracted with EtOAc (45 mL). The combined organic layers were washed with brine (5 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 1/0 to 9/1) to give desired 3-chloro-2-(2, 6-dimethylmorpholin-4-yl) aniline (200 mg, 831 pmol, 75.0% yield) as a brown oil. MS (ESI): mass calcd. For C12H17CIN2O 240.10, m/z found 241.0 [M+H] ⁺ .

[00504] Step 3. To a solution of 3-chloro-2-(2, 6-dimethylmorpholin-4-yl) aniline (200 mg, 831 pmol, 1 eq) in Py (5 mL) was added 5 -(dimethylsulfamoyl) thiophene -2-sulfonyl chloride (265 mg, 914 pmol, 1.1 eq). The mixture was stirred at 15 °C for 12 hours. LCMS showed 3-chloro-2-(2, 6- dimethylmorpholin-4-yl) aniline was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a crude oil. The residue was purified by prep-TLC (SiC>2, petroleum ether/EtOAc = 3/1). The purified product was dissolved in 20 mL of water, and then the solution was lyophilized to give desired N5-[3-chloro-2-(2, 6-dimethylmorpholin-4- yl)phenyl]-N2, N2-dimethyl-thiophene-2, 5-disulfonamide (109 mg, 215 pmol, 25.9% yield, 97.6% purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.79 (s, 1H), 7.76 - 7.69 (m, 2H), 7.41 - 7.32 (m, 1H), 7.20 - 7.12 (m, 1H), 7.05 - 6.98 (m, 1H), 4.14 - 3.99 (m, 2H), 3.41 (br s, 1H), 3.34 - 3.21 (m, 1H), 2.89 (br d, J= 17.1 Hz, 2H), 2.69 (s, 6H), 1.25 - 1.16 (m, 6H). HPLC: 97.59% (220 nm), 97.63% (215 nm), 97.76% (254 nm). MS (ESI): mass calcd. For C18H24CIN3O5S3493.06, m/z found 494.2 [M+H] ⁺ .

Example 101: N2-(2-(3-hydroxypyrrolidin-l-yl)-3-methylphenyl)-N5, N5-dimethylthiophene-2, 5-disulfonamide

[00505] Step 1. To a solution of 2-fluoro-l-methyl-3 -nitro-benzene (356 mg, 2.30 mmol, 1 eq) and pyrrolidin-3-ol (200 mg, 2.30 mmol, 185 μL, 1 eq) in dioxane (5 mL) was added K ₂ CO ₃ (635 mg, 4.59 mmol, 2 eq). The mixture was stirred at 100 °C for 12 hours. LCMS showed 2-fluoro-l- methyl-3 -nitro-benzene was consumed and desired mass was detected. The crude was added H2O (10 mL), and extracted with EtOAc 45 mL (15 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/Ethyl acetate=2/l to 1/1) to give desired 1 -(2-methyl-6-nitro-phenyl) pyrrolidin -3-ol (300 mg, 1.35 mmol, 58.8% yield) as a brown oil. MS (ESI): mass calcd. For C11H14N2O3 222.10, m/z found 223.0 [M+H] ⁺ .

[00506] Step 2: To a solution of l-(2-methyl-6-nitro-phenyl) pyrrolidin -3-ol (300 mg, 1.35 mmol, eq) in MeOH (5 mL) was added H2 and Pd/C (30 mg, 10% purity). The mixture was stirred at 15 °C for 2 hours. LCMS showed 1 -(2-methyl-6-nitro-phenyl) pyrrolidin -3-ol was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired l-(2-amino-6-methyl-phenyl) pyrrolidin-3-ol (250 mg, crude) as a brown oil. MS (ESI): mass calcd. For C11H16N2O 192.13, m/z found 193.4 [M+H] ⁺ .

[00507] Step 3. To a solution of 1 -(2-amino-6-methyl-phenyl) pyrrolidin-3-ol (250 mg, 1.30 mmol, 1 eq) in Py (5 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (377 mg, 1.30 mmol, 1 eq). The mixture was stirred at 15 °C for 12 hours. LCMS showed l-(2-amino-6- methyl-phenyl) pyrrolidin-3-ol was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a crude oil. The residue was purified by prep- HPLC (column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (lOmM NH ₄ HCO ₃ )-ACN]; B%: 35%-65%, 8 min). The purified product was dissolved in 20 mL of water, and then the solution was lyophilized to give desired N2-(2-(3-hydroxypyrrolidin-l-yl)-3- methylphenyl)-N5, N5 -dimethylthiophene-2, 5 -disulfonamide (39.1 mg, 87.5 pmol, 6.73% yield, 99.7 % purity) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 7.69 - 7.61 (m, 2H), 7.38 - 7.32 (m, 1H), 7.14 - 7.07 (m, 1H), 7.02 - 6.95 (m, 1H), 4.35 - 4.28 (m, 1H), 3.35 - 3.31 (m, 1H), 3.20 - 3.13 (m, 1H), 2.99 - 2.83 (m, 2H), 2.64 (s, 6H), 2.61 - 2.54 (m, 2H), 2.21 (s, 3H), 2.11 - 1.97 (m, 1H), 1.90 - 1.80 (m, 1H). HPLC: 99.68% (220 nm), 99.48% (215 nm), 99.94% (254 nm). MS (ESI): mass calcd. For C17H23N3O5S3445.08 m/z found 446.1 [M+H] ⁺ .

Example 102: 5-(tert-butylthio)-N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene- 2-sulfonamide

[00508] Step 1. To a solution of 5-bromo-N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene-2-sulfonamide (200 mg, 431 pmol, 1 eq), 2-methylpropane-2-thiol (50.6 mg, 561 pmol, 63.1 μL, 1.3 eq), BINAP (268 mg, 431 pmol, 1 eq) and CS ₂ CO ₃ (351 mg, 1.08 mmol, 2.5 eq) in Tol. (2 mL) was added Pd2(dba)3 (197 mg, 216 pmol, 0.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 5-bromo-N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene-2- sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was separated between 5 mL of water and 8 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by column chromatography (SiCF, petroleum ether/EtOAc = 100/1 to 50/1) to give desired 5-(tert-butylthio)-N-(3-chloro-2-(4, 4- dimethylpiperidin-l-yl) phenyl) thiophene-2-sulfonamide (140 mg, 296 pmol, 68.6% yield) as a yellow oil. MS (ESI): mass calcd. For C21H29CIN2O2S3 472.11 m/z found 473.1 [M+H] ⁺ .

[00509] Step 2'. To a solution of 5-(tert-butylthio)-N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene-2-sulfonamide (140 mg, 296 pmol, 1 eq} in DCM (3 mL) was added m-CPBA (180 mg, 888 pmol, 85% purity, 3 eq} at 0 °C. The mixture was stirred at 20 °C for 12 hours. LC- MS showed 5-(tert-butylthio)-N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) thiophene-2- sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was partitioned between 5 mL of sat. Na ₂ SO ₃ and 10 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of sat. Na ₂ SO ₃ , ImL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep- HPLC (column: Waters Xbridge BEH C18 100*30mm*10um;mobile phase: [water(10mM NILHCO3)-ACN];B%: %-%,10min) to give desired 5-(tert-butylthio)-N-(3-chloro-2-(4, 4- dimethylpiperidin-l-yl) phenyl) thiophene-2-sulfonamide (27.8 mg, 52.2 pmol, 17.6% yield, 94.9% purity) as a yellow solid. ¹ H NMR (400MHz, DMSO-rfc) δ 9.58 (s, 1H), 7.77 (d, J= 4.0 Hz, 1H), 7.69 (d, J= 4.0 Hz, 1H), 7.30 - 7.24 (m, 2H), 7.22 - 7.15 (m, 1H), 3.25 (br s, 2H), 2.33 (br s, 2H), 1.59 - 1.38 (m, 2H), 1.37 - 1.28 (m, 2H), 1.26 (s, 9H), 0.96 (br s, 6H). HPLC: 94.85% (220 nm), 95.77% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C21H29CIN2O4S3 504.10 m/z found 505.1 [M+H] ⁺ .

Example 103: N2-(2-([l, 4'-bipiperidin]-l'-yl)-3-chlorophenyl)-N5, N5-dimethylthiophene-2, 5- disulfonamide

[00510] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (1 g, 5.70 mmol, 1 eq} and 1- (4-piperidyl)piperidine (1.52 g, 7.41 mmol, 1.3 eq, HC1) in dioxane (15 mL) was added K ₂ CO ₃ (2.36 g, 17.1 mmol, 3 eq}. The mixture was stirred at 100 °C for 12 hours. LC-MS showed l-chloro-2- fluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCfi, petroleum ether/EtOAc = 1/0 to 2/1) to give desired l-(2-chloro-6-nitro-phenyl)-4-(l -piperidyl) piperidine (1.8 g, crude) as a yellow oil. MS (ESI): mass calcd. For C16H22CIN3O2 323.14, m/z found 324.3 [M+H] ⁺ .

[00511] Step 2\ To a solution of l-(2-chloro-6-nitro-phenyl)-4-(l -piperidyl) piperidine (1.8 g, 5.56 mmol, 1 eq} in CH3COOH (10 mL) was added Fe (931 mg, 16.7 mmol, 3 eq}. The mixture was stirred at 80 °C for 1 hour. TLC (EtOAc/Methanol=5/l) showed l-(2-chloro-6-nitro-phenyl)-4-(l- piperidyl) piperidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was added to water (20 mL), extracted with DCM (60 mL). The combined organic layers were washed with brine (20 mL) and dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiC>2, petroleum ether/EtOAce=l/0 to 2/1) to give desired 3-chloro-2-[4-(l-piperidyl)-l-piperidyl] aniline (165 mg, 562 pmol, 10.1% yield) as ayellow oil.

[00512] Step 3. To a solution of 3 -chloro-2-[4-(l -piperidyl)-! -piperidyl] aniline (140 mg, 476 pmol, 1 eq} in Py (1.5 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (207 mg, 715 pmol, 1.5 eq}. The mixture was stirred at 15 °C for 2 hours. LC-MS showed 3-chloro-2-[4-(l- piperidyl)-! -piperidyl] aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, EtOAc/Methanol = 5/1) to give desired N2- (2-([l, 4'-bipiperidin]-l'-yl)-3-chlorophenyl)-N5, N5 -dimethylthiophene-2, 5 -disulfonamide (18 mg, 31.6 pmol, 6.63% yield, 96.1% purity) as a faint yellow solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 7.67 - 7.60 (m, 1H), 7.62 - 7.17 (m, 2H), 7.12 (br d, J= 6.5 Hz, 1H), 6.86 - 6.58 (m, 2H), 3.20 - 2.99 (m, 5H), 2.76 (br d, J= 8.4 Hz, 3H), 2.63 (s, 6H), 1.93 (br s, 3H), 1.72 (br s, 6H), 1.54 (br s, 2H). HPLC: 99.68% (220 nm), 99.48% (215 nm), 99.94% (254 nm). MS (ESI): mass calcd. For C22H31CIN4O4S3 546.12 m/z found 547.2 [M+H] ⁺ .

Example 104: N2, N2-dimethyl-N5-(3-methyl-2-(3-phenylazetidin-l-yl) phenyl) thiophene-2, 5- disulfonamide

[00513] Step 1. To a solution of Zn (6.41 g, 98.0 mmol, 4 eq) in DMA (15 mL) was added 1, 2- dibromoethane (691 mg, 3.68 mmol, 277 μL, 0.15 eq) and TMSC1 (799 mg, 7.35 mmol, 933 μL, 0.3 eq) at 25 °C. After addition, the mixture was stirred at this temperature for 25 mins, and then tertbutyl 3 -iodoazetidine- 1 -carboxylate (17.4 g, 61.3 mmol, 2.5 eq) in DMA (15 mL) was added dropwise at below 65 °C. The resulting mixture was stirred at 25 °C for 1 hours. To a solution of iodobenzene (5 g, 24.5 mmol, 2.73 mL, 1 eq) in DMA (50 mL) was added Cui (607 mg, 3.19 mmol, 0.13 eq) and Pd(dppf)C12.CH ₂ C12 (1.20 g, 1.47 mmol, 0.06 eq) at 25 °C. After addition, the mixture was stirred at this temperature for 5 mins, and the solution containing the organozinc in DMA generated above was added drop wise at 25 °C. The resulting mixture was stirred at 80 °C for 0.5 hours. TLC (petroleum ether/Ethyl acetate = 5:1) indicated iodobenzene was consumed completely and three new spots formed. Then it was separated between 100 mL of water and 200 mL of ethyl acetate. The organic phase was separated, washed with 100 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0~5% Ethyl acetate/petroleum ether gradient @ 100 mL/min) to give desired tert-butyl 3 -phenylazetidine- 1- carboxylate (6 g, crude) as a yellow oil.

[00514] Step 2'. A mixture of tert-butyl 3 -phenylazetidine- 1 -carboxylate (2.5 g, 10.7 mmol, 1 eq) in HCl/MeOH (4 M, 25.0 mL, 9.33 eq) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 12 hours under N2 atmosphere. TLC (petroleum ether/Ethyl acetate = 3/1) indicated was tert-butyl 3 -phenylazetidine- 1 -carboxylate consumed completely and one new spot formed. The reaction mixture was concentrated in vacuum to give desired 3 -phenylazetidine (2.2 g, crude, HC1) as a yellow oil.

[00515] Step 3: A mixture of 3 -phenylazetidine (2.2 g, 13.0 mmol, 1 eq, HC1), 2-fluoro-l-methyl- 3 -nitro-benzene (2.41 g, 15.6 mmol, 1.2 eq) and CS ₂ CO ₃ (10.6 g, 32.4 mmol, 2.5 eq) in DMSO (25 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100 °C for 12 hours under N2 atmosphere. LC-MS showed 3 -phenylazetidine was consumed completely and one main peak with desired mass was detected. Then it was separated between 50 mL of water and 100 mL of ethyl acetate. The organic phase was separated, washed with 70 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-10% Ethyl acetate/petroleum ether gradient @ 80 mL/min) to give desired l-(2-methyl-6- nitrophenyl)-3 -phenylazetidine (1.9 g, 7.08 mmol, 54.3% yield) as a yellow oil. MS (ESI): mass calcd. For C16H16N2O2 268.12, m/z found 269.2 [M+H] ⁺ .

[00516] Step 4. A mixture of l-(2-methyl-6-nitrophenyl)-3 -phenylazetidine (0.9 g, 3.35 mmol, 1 eq), Fe (937 mg, 16.8 mmol, 5 eq) and NH4CI (897 mg, 16.8 mmol, 5 eq) in H ₂ O (2 mL) and EtOH (9 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 70 °C for 12 hours under N2 atmosphere. LC-MS showed l-(2-methyl-6-nitrophenyl)-3 -phenylazetidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove MeOH. The reaction mixture was partitioned between 10 mL of H ₂ O and 20 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 3-methyl-2-(3-phenylazetidin-l-yl)aniline (0.7 g, crude) as a yellow oil. MS (ESI): mass calcd. For C16H18N2 238.15, m/z found 239.2 [M+H] ⁺ .

[00517] Step 5: A mixture of 3-methyl-2-(3-phenylazetidin-l-yl) aniline (0.3 g, 1.26 mmol, 1 eq), 5-(N, N-dimethylsulfamoyl)thiophene-2-sulfonyl chloride (365 mg, 1.26 mmol, 1 eq) and TEA (318 mg, 3.15 mmol, 438 μL, 2.5 eq) in DCM (3 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 12 hours under N2 atmosphere. LC-MS showed 3-methyl-2- (3-phenylazetidin-l-yl) aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was partitioned between 3 mL of water and 5 mL of DCM. The organic phase was separated, washed three times with 6 mL of water and 2 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini-NX 80*40mm*3um; mobile phase: [water (lOmM NILHCOsj-ACN]; B%: 30%-60%,8 min) to give desired N2, N2-dimethyl-N5-(3-methyl-2-(3- phenylazetidin-l-yl) phenyl)thiophene-2, 5-disulfonamide (58.4 mg, 116 pmol, 9.25% yield, 98.0% purity) as a yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.83 (s, 1H), 7.67 (d, J= 4.0 Hz, 1H), 7.53 (d, J= 4.0 Hz, 1H), 7.45 - 7.40 (m, 2H), 7.39 - 7.33 (m, 2H), 7.28 - 7.22 (m, 1H), 6.95 (d, J = 7.2 Hz, 1H), 6.49 (t, J= 7.6 Hz, 1H), 6.24 (d, J= 7.2 Hz, 1H), 4.59 (t, J= 7.8 Hz, 2H), 4.16 (t, J = 6.9 Hz, 2H), 3.74 (quin, J= 7.2 Hz, 1H), 2.68 (s, 6H), 2.25 (s, 3H). HPLC: 98.03% (220 nm), 98.09% (215 nm), 98.37% (254 nm). MS (ESI): mass calcd. For C22H25N3O4S3 491.10 m/z found 492.1 [M+H] ⁺ .

Example 105: N2-(3-chloro-2-(3-isopropylpyrrolidin-l-yl) phenyl)-N5, N5-dimethylthiophene- 2, 5-disulfonamide

[00518] Step 1. A mixture of potassium;2-methylpropan-2-olate (3.20 g, 28.5 mmol, 2.5 eq), isopropyl(triphenyl)phosphonium;iodide (12.3 g, 28.5 mmol, 2.5 eq) in tetrahydrofuran (30 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 1 hour under N2 atmosphere. Then l-benzylpyrrolidin-3-one (2 g, 11.4 mmol, 1.87 mL, 1 eq) in tetrahydrofuran (10 mL) was added and the mixture was 35 °C for 4.5 hours. TLC (petroleum ether/Ethyl acetate=3/l) indicated no l-benzylpyrrolidin-3-one remained and one major new spot with lower polarity was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H ₂ O 100 mL and extracted with EtOAc 600 mL (200 mL * 3). The combined organic layers were washed with brine 300 mL (100 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0-15% Ethyl acetate/petroleum ether gradient @ 60 mL/min) to give desired l-benzyl-3-isopropylidene- pyrrolidine (600 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C14H19N 201.15, m/z found 202.3 [M+H] ⁺ .

[00519] Step 2'. To a solution of l-benzyl-3-isopropylidene-pyrrolidine (600 mg, 2.98 mmol, 1 eq) in formic acid (1 mL) and MeOH (5 mL) was added Pd/C(10%, 0.1 g) and Pd(OH)2/C (10%, 0.1 g) under N2 atmosphere. The suspension was degassed and purged with H2 for 3 times. The mixture was stirred under H2 (50 Psi or atm.) at 30 °C for 48 hours. LC-MS indicated l-benzyl-3- isopropylidene-pyrrolidine was consumed completely. The reaction was clean according to TLC (petroleum ether/Ethyl acetate=3:l). The reaction mixture was filtered and washed 3 times with EtOAc and concentrated under reduced pressure to give desired 3-isopropylpyrrolidine (300 mg, crude) as a yellow oil which was used directly for the next step without further purification. MS (ESI): mass calcd. For C7H15N 113.12, m/z found 114.3 [M+H] ⁺ .

[00520] Step 3. To a solution of l-chloro-2-fluoro-3 -nitro-benzene (200 mg, 1.14 mmol, 1 eq) in DMSO (3 mL) was added CS ₂ CO ₃ (928 mg, 2.85 mmol, 2.5 eq) and 3-isopropylpyrrolidine (193 mg, 1.71 mmol, 1.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed l-chloro-2- fluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O 10 mL and extracted with EtOAc 60 mL (20 mL * 3). The combined organic layers were washed with brine 15 mL (5 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-10% Ethyl acetate/petroleum ether gradient @ 35 mL/min) to give desired 1- (2-chloro-6-nitro-phenyl)-3-isopropyl-pyrrolidine (140 mg, 521 pmol, 45.7% yield) as a yellow oil. MS (ESI): mass calcd. For C13H17CIN2O2 268.10, m/z found 269.2 [M+H] ⁺ .

[00521] Step 4. To a solution of l-(2-chloro-6-nitro-phenyl)-3-isopropyl-pyrrolidine (140 mg, 521 pmol, 1 eq) in EtOH (3 mL) and H ₂ O (0.6 mL) was added Fe (291 mg, 5.21 mmol, 10 eq) and NH4CI (279 mg, 5.21 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed l-(2-chloro-6-nitro-phenyl)-3-isopropyl-pyrrolidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (5 mL) and extracted with EtOAc (20 mL * 3). The combined organic layers were washed with brine (15 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-10% Ethyl acetate/petroleum ether gradient @ 30 mL/min) to give desired 3-chloro-2-(3-isopropylpyrrolidin-l-yl) aniline (80 mg, 335 pmol, 64.3% yield) as a yellow oil. MS (ESI): mass calcd. For C13H19CIN2 238.12, m/z found 239.2 [M+H] ⁺ .

[00522] Step 5: To a solution of 3-chloro-2-(3-isopropylpyrrolidin-l-yl) aniline (80 mg, 335 pmol, 1 eq) in pyridine (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (117 mg, 402 pmol, 1.2 eq). The mixture was stirred at 20 °C for 3 hours. LC-MS showed 3-chloro-2-(3- isopropylpyrrolidin-l-yl) aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition: column: Waters Xbridge BEH Cl 8 100*25mm*5um; mobile phase: [water (10mM NELHCO3)-ACN]; B%: 45%-80%, 10 min) to give desired N2-(3-chloro-2-(3-isopropylpyrrolidin-l-yl)phenyl)-N5, N5 -dimethylthiophene-2, 5- disulfonamide (62 mg, 124 pmol, 37.0% yield, 98.3% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 9.91 (s, 1H), 7.66 (q, J= 4.0 Hz, 2H), 7.42 (dd, J= 1.5, 7.9 Hz, 1H), 7.34 - 7.19 (m, 2H), 3.10 - 2.99 (m, 1H), 2.85 (q, J= 7.3 Hz, 2H), 2.74 - 2.68 (m, 1H), 2.66 (s, 6H), 2.07 - 1.89 (m, 2H), 1.70 - 1.58 (m, 1H), 1.52 - 1.39 (m, 1H), 0.93 (d, J= 6.6 Hz, 3H), 0.84 (d, J= 6.5 Hz, 3H). HPLC: 98.28% (220 nm), 98.60% (215 nm), 98.41% (254 nm). MS (ESI): mass calcd. For C19H26CIN3O4S3491.08 m/z found 492.1 [M+H] ⁺ .

Example 106: N-(3-chloro-2-(4, 4-dimethylpiperidin-l-yl) phenyl) -5-isobutyrylthiophene-2- sulfonamide

[00523] Step 1. To a solution of 5-bromo-N-[3-chloro-2-(4, 4-dimethyl-l- piperidyl)phenyl]thiophene-2-sulfonamide (110 mg, 237 pmol, 1 eq) and N-methoxy-N, 2-dimethyl- propanamide (65.3 mg, 498 pmol, 2.1 eq) in THF (2 mL) was added w-BuLi (2.5 M, 94.9 μL, 1 eq) at -70 °C. The mixture was stirred at -70 °C for 0.5 hour. LC-MS showed 5-bromo-N-[3-chloro-2- (4, 4-dimethyl-l -piperidyl) phenyl] thiophene-2-sulfonamide was consumed completely and desired mass was detected. Then it was separated between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep- HPLC (neutral condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (lOrnM NH4HCOs)-ACN]; B%: 55%-85%, 8min) to give desired N-(3-chloro-2-(4, 4- dimethylpiperidin-l-yl) phenyl) -5-isobutyrylthiophene-2-sulfonamide (3 mg, 6.59 pmol, 2.78% yield, 100% purity) as a pale yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.40 (br s, 1H), 7.97 (br d, J= 3.5 Hz, 1H), 7.63 (br d, J= 2.1 Hz, 1H), 7.39 - 7.17 (m, 3H), 3.56 - 3.42 (m, 3H), 2.33 - 2.28 (m, 1H), 1.56 - 1.43 (m, 2H), 1.36 - 1.19 (m, 3H), 1.11 (s, 3H), 1.09 (s, 3H), 0.96 (br s, 6H). HPLC: 100.00% (220 nm), 100.00% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C21H27CIN2O3S2454.12 m/z found 455.1 [M+H] ⁺ .

Example 107: 5-[[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] methylsulfonyl] -N, N- dimethyl-thiophene-2-sulfonamide

[00524] Step 1. To a solution of 5-(N, N-dimethylsulfamoyl) thiophene-2-sulfonyl chloride (290 mg, 1.00 mmol, 1 eq) in Tol. (20 mL) was added PPhs (787 mg, 3.00 mmol, 3 eq) at 0 °C. The mixture was allowed to warm to 20 °C and stirred for 2 hours. To the above solution was added H2O (3 mL) and the mixture was stirred for another 0.17 hours. LC-MS showed 5-(N, N- dimethylsulfamoyl) thiophene-2-sulfonyl chloride was consumed completely and one main peak with desired mass was detected. The reaction mixture was partitioned between Ethyl acetate 30 mL and water 50 mL. The organic phase was separated, washed with Ethyl acetate 30 mL (10 mL * 3), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-50% Ethyl acetate/petroleum ether gradient @ 40 mL/min) to give desired 5-mercapto-N, N- dimethylthiophene-2-sulfonamide (500 mg, crude) as a yellow solid. MS (ESI): mass calcd. For C6H9NO2S3 222.98 m/z found 224.1 [M+H] ⁺ .

[00525] Step 2\ A mixture of 3-chloro-2-fluorobenzaldehyde (10 g, 63.1 mmol, 1 eq), 4, 4- dimethylpiperidine (10.4 g, 69.4 mmol, 1.1 eq, HC1) and CS ₂ CO ₃ (51.4 g, 158 mmol, 2.5 eq) in DMSO (100 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 120 °C for 12 hours under N2 atmosphere. TLC (petroleum ether/Ethyl acetate = 5/1) indicated 3- chloro-2-fluorobenzaldehyde was consumed completely and two new spots formed. Then it was separated between 50 mL of water and 100 mL of ethyl acetate. The organic phase was separated, washed with 70 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The crude product was triturated with ethyl acetate at 20 °C for 15 mins. The mixture was filtered and the filter cake was dried in vacuum to give desired 3-chloro-2-(4, 4- dimethyl-1 -piperidyl) benzaldehyde (4 g, 15.9 mmol, 25.2% yield) as a white solid.

[00526] Step 3. To a solution of 3-chloro-2-(4, 4-dimethyl-l -piperidyl) benzaldehyde (630 mg, 2.50 mmol, 1 eq) in MeOH (7 mL) was added NaBEL (189 mg, 5.00 mmol, 2 eq) in 20 portions at 0 °C. After addition, the mixture was stirred at 20 °C for 1 hour. LC-MS showed 3-chloro-2-(4, 4- dimethyl-1 -piperidyl) benzaldehyde was consumed completely and one main peak with desired mass was detected. The reaction mixture was partitioned between 20 mL of HC1 (IM) and ethyl acetate (60 mL). The organic phase was separated, washed with brine (40 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired [3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] methanol (0.52 g, crude) as a yellow solid. MS (ESI): mass calcd. For C14H20CINO 253.12 m/z found 254.2 [M+H] ⁺ .

[00527] Step 4. To a solution of [3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] methanol (100 mg, 394 pmol, 1 eq) and TEA (59.8 mg, 591 pmol, 82.3 μL, 1.5 eq) in DCM (1 mL) was added MsCl (67.7 mg, 591 pmol, 45.8 μL, 1.5 eq) at 0 °C. The mixture was stirred at 0 °C for 1 hour. TLC (petroleum ether/Ethyl acetate = 3/1) indicated [3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] methanol was consumed completely and one new spot formed. Then it was separated between 2 mL of water and 4 mL of DCM. The organic phase was separated, washed with 3 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired [3-chloro-2-(4, 4- dimethyl-1 -piperidyl) phenyl] methyl methanesulfonate (150 mg, crude) as ayellow oil.

[00528] Step 5: To a solution of [3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] methyl methanesulfonate (150 mg, 452 pmol, 1 eq) in DMF (2 mL) was added 5-mercapto-N, N- dimethylthiophene-2-sulfonamide (202 mg, 904 pmol, 2 eq) and CS ₂ CO ₃ (442 mg, 1.36 mmol, 3 eq). The mixture was stirred at 90 °C for 1 hour. LC-MS showed [3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] methyl methanesulfonate was consumed completely and one main peak with desired mass was detected. Then it was separated between 10 mL of water and 20 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by column chromatography (SiCL, petroleum ether/Ethyl acetate = 1/0 to 10/1) to give desired 5-((3-chloro-2-(4, 4-dimethylpiperidin-l-yl) benzyl) thio)-N, N-dimethylthiophene-2-sulfonamide (150 mg, 327 pmol, 72.3% yield) as a yellow solid. MS (ESI): mass calcd. For C20H27CIN2O2S3 458.09 m/z found 459.1 [M+H] ⁺ .

[00529] Step 6. To a solution of 5-((3-chloro-2-(4, 4-dimethylpiperidin-l-yl) benzyl) thio)-N, N- dimethylthiophene-2-sulfonamide (150 mg, 327 pmol, 1 eq) in DCM (2 mL) was added m-CPBA (99.5 mg, 490 pmol, 85% purity, 1.5 eq) at 0 °C. The mixture was stirred at 20 °C for 12 hours. LC- MS showed 5-((3-chloro-2-(4, 4-dimethylpiperidin-l-yl) benzyl) thio)-N, N-dimethylthiophene-2- sulfonamide was consumed completely and one main peak with desired mass was detected. Then it was partitioned between 5 mL of sat. Na ₂ SO ₃ and 10 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of sat. Na ₂ SO ₃ , 1 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep- TLC (SiC>2, petroleum ether/Ethyl acetate = 3/1) to give desired 5-[[3-chloro-2-(4, 4-dimethyl-l- piperidyl) phenyl] methylsulfonyl] -N, N-dimethyl-thiophene-2-sulfonamide (44.2 mg, 89.8 pmol, 27.5% yield, 99.7% purity) as a white solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 7.83 (d, J= 4.0 Hz, 1H), 7.76 (d, J= 3.9 Hz, 1H), 7.45 (dd, J= 1.5, 7.8 Hz, 1H), 7.34 - 7.30 (m, 1H), 7.28 - 7.23 (m, 1H), 5.05 (s, 2H), 3.45 (br t, J= 11.0 Hz, 2H), 2.68 (s, 6H), 2.25 (br d, J= 10.0 Hz, 2H), 1.47 (dt, J= 4.3, 12.4 Hz, 2H), 1.28 (br d, J= 12.6 Hz, 2H), 0.97 (s, 6H). HPLC: 99.72% (220 nm), 99.52% (215 nm), 99.84% (254 nm). MS (ESI): mass calcd. For C20H27CIN2O4S3490.08 m/z found 491.1 [M+H] ⁺ .

Example 108: 5-[[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] -fluoro-methyl] sulfonyl-N, N- dimethyl-thiophene-2-sulfonamide [00530] Step 1. To a solution of 5-[[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl] methylsulfonyl] -N, N-dimethyl-thiophene-2-sulfonamide (50 mg, 102 pmol, 1 eq) in Tol. (1 mL) was added dropwise LDA (2 M, 66.2 μL, 1.3 eq) at -70 °C. After addition, the mixture was stirred at this temperature for 0.5 hour, and then NFSI (38.5 mg, 122 pmol, 1.2 eq) was added dropwise at -70 °C. The resulting mixture was stirred at -70 °C for 1 hour. LC-MS showed 5-[[3-chloro-2-(4, 4- dimethyl-1 -piperidyl) phenyl] methylsulfonyl] -N, N-dimethyl-thiophene-2-sulfonamide was consumed completely and one main peak with desired mass was detected. The mixture was added dropwise NH4CI for at 0 °C, and extracted three times with 6 mL of Ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep-WY,C (column: Welch Xtimate C18 100*25mm*3um; mobile phase: [water (0.04%HCl)-ACN]; B%: 20%-70%, 8min). The reaction mixture was partitioned between 5 mL of NaOH (IN) and 10 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 5-[[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl]-fluoro-methyl]sulfonyl-N,N-dimethyl-thiophene-2-sulf onamide (16.5 mg, 32.4 pmol, 31.8% yield, 99.9% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.01 (d, J= 4.0 Hz, 1H), 7.88 (d, J= 4.0 Hz, 1H), 7.66 - 7.62 (m, 1H), 7.41 - 7.36 (m, 1H), 7.35 - 7.32 (m, 1H), 7.20 (s, 1H), 3.64 - 3.46 (m, 2H), 2.76 (s, 6H), 2.71 - 2.60 (m, 2H), 1.61 (br t, J= 12.4 Hz, 2H), 1.39 - 1.22 (m, 2H), 1.00 (d, J= 12.0 Hz, 6H). HPLC: 99.93% (220 nm), 99.75% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C20H26CIFN2O4S3 508.07 m/z found 509.1 [M+H] ⁺ .

Example 109: N5-[3-chloro-2-[2-(methoxymethyl)-l-piperidyl] phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00531] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (500 mg, 2.85 mmol, 1 eq) in DMSO (4 mL) was added CS ₂ CO ₃ (1.86 g, 5.70 mmol, 2 eq) and 2-(methoxymethyl) piperidine (566 mg, 3.42 mmol, 1.2 eq, HC1). The mixture was stirred at 100 °C for 12 hours. LC-MS showed desired mass was detected. The reaction mixture was added to water (300 mL), the aqueous phase was extracted with EtOAc (150 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 1/0 to 1/1) to give desired l-(2-chloro-6-nitro-phenyl)-2- (methoxymethyl) piperidine (220 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C13H17CIN2O3 284.09, m/z found 285.0 [M+l] ⁺ .

[00532] Step 2\ To a solution of l-(2-chloro-6-nitro-phenyl)-2-(methoxymethyl) piperidine (220 mg, 773 pmol, 1 eq) in H ₂ O (0.1 mL) and EtOH (0.5 mL) was added Fe (345 mg, 6.18 mmol, 8 eq) and NH4CI (413 mg, 7.73 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LCMS showed l-(2-chloro-6-nitro-phenyl)-2-(methoxymethyl) piperidine was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The reaction mixture was added to water (20 mL), the aqueous phase was extracted with EtOAc (30 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 5/1) to give desired 3 -chloro-2-[2-(methoxymethyl)-l -piperidyl] aniline (100 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C13H19CIN2O 254.12, m/z found 255.3 [M+l] ⁺ . [00533] Step 3. To a solution of 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (171 mg, 589 pmol, 1.5 eq) in Py (1 mL) was added 3 -chloro-2-[2-(methoxymethyl)-l -piperidyl] aniline (100 mg, 393 pmol, 1 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed 5- (dimethylsulfamoyl) thiophene-2-sulfonyl chloride was consumed completely and desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 2/1) to give desired N5-[3-chloro-2-[2-(methoxymethyl)- 1 -piperidyl] phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (19.8 mg, 38.4 pmol, 9.79% yield, 98.6% purity) as an off-white solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 9.01 (s, 1 H) 7.81 (d, J = 4.03 Hz, 1 H) 7.70 (d, J= 4.03 Hz, 1 H) 7.60 - 7.56 (m, 1 H) 7.36 (t, J= 8.13 Hz, 1 H) 7.29 - 7.25 (m, 1 H) 3.55 - 3.45 (m, 1 H) 3.22 - 3.14 (m, 1 H) 3.04 (dd, J= 10.09, 2.26 Hz, 1 H) 2.94 (s, 3 H) 2.76 (dd, J= 10.02, 3.91 Hz, 1 H) 2.65 (s, 6 H) 1.80 (br d, J= 10.15 Hz, 1 H) 1.71 - 1.57 (m, 3 H) 1.54 - 1.44 (m, 1 H) 1.42 - 1.25 (m, 2 H). HPLC: 98.63% (220 nm), 99.34% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C19H26CIN3O5S3 507.07 m/z found 508.0 [M+H] ⁺ .

Example 110: N5-[2-(9-azabicyclo [3.3.1] nonan-9-yl)-3-chloro-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide [00534] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (200 mg, 1.14 mmol, 1 eq) and 9-azabicyclo[3.3.1] nonane (221 mg, 1.37 mmol, 1.2 eq, HC1) in dioxane (4 mL) was added K ₂ CO ₃ (315 mg, 2.28 mmol, 2 eq) and DIEA (295 mg, 2.28 mmol, 397 μL, 2 eq). The mixture was stirred at 120°C for 24 hours. LC-MS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 10/1) to give desired 9-(2-chloro-6-nitro-phenyl)-9-azabicyclo [3.3.1] nonane (230 mg, 81 pmol, 71.9% yield) as ayellow oil. MS (ESI): mass calcd. For C14H17CIN2O2280.10, m/z found 281.3 [M+l] ⁺ . [00535] Step 2'. To a solution of 9-(2-chloro-6-nitro-phenyl)-9-azabicyclo [3.3.1] nonane (300 mg, 1.07 mmol, 1 eq) in H ₂ O (0.5 mL) and EtOH (3 mL) was added Fe (477mg, 8.55 mmol, 8 eq) and NH4CI (857 mg, 16.0 mmol, 15 eq). The mixture was stirred at 80°C for 3 hours. LC-MS showed 9-(2-chloro-6-nitro-phenyl)-9-azabicyclo [3.3.1] nonane was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was added H ₂ O (30 mL) and extracted with EtOAc (90 mL). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether/EtOAc = 5/1) to give desired 2-(9-azabicyclo [3.3.1] nonan-9-yl)-3 -chloro-aniline (200 mg, 798 pmol, 74.6% yield) as a red oil. MS (ESI): mass calcd. For C14H19CIN2 250.12, m/z found 251.3 [M+l] ⁺ .

[00536] Step 3. To a solution of 2-(9-azabicyclo [3.3.1] nonan-9-yl)-3 -chloro-aniline (200 mg, 798 pmol, 1 eq) in Py (1 mL) was added 5-(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (347 mg, 1.20 mmol, 1.5 eq). The mixture was stirred at 15 °C for 1.5 hours. LC-MS showed 2-(9- azabicyclo [3.3.1] nonan-9-yl)-3 -chloro-aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WLC (neutral condition; column: Phenomenex Gemini-NX 80*40mm*3um;mobile phase: [water (10 mM NEEHCO3)-ACN]; B%: 55% - 85%, 8 min) to give desired N5-[2-(9-azabicyclo [3.3.1] nonan-9-yl)-3 -chloro-phenyl] -N2, N2- dimethyl-thiophene-2, 5 -disulfonamide (149 mg, 291 pmol, 36.5% yield, 98.7% purity) as a pale yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.50 (s, 1 H) 7.80 (d, J= 3.91 Hz, 1 H) 7.73 (d, J = 3.91 Hz, 1 H) 7.33 - 7.24 (m, 1 H) 7.14 - 7.09 (m, 2 H) 3.68 (br s, 2 H) 2.69 (s, 6 H) 2.46 - 2.38 (m, 2 H) 1.85 - 1.70 (m, 6 H) 1.66 (br d, J= 6.97 Hz, 2 H) 1.55 - 1.45 (m, 2 H). HPLC: 98.66% (220 nm), 98.50% (215 nm), 99.43% (254 nm). MS (ESI): mass calcd. For C20H26CIN3O4S3 503.08 m/z found 504.9 [M+H] ⁺ .

Example 111: N2-[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] -N5-methyl- thiophene-2, 5- disulfonamide

111

[00537] Step 1. To a solution of 5-bromothiophene-2-sulfonyl chloride (2 g, 7.65 mmol, 1 eq) in DCM (20 mL) was added methanamine; hydrochloride (568 mg, 8.41 mmol, 1.1 eq) and TEA (1.93 g, 19.12 mmol, 2.66 mL, 2.5 eq). The mixture was stirred at 15 °C for 2 hours. LCMS showed 5- bromothiophene-2-sulfonyl chloride was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired 5- bromo-N-methyl-thiophene-2-sulfonamide (2.05 g, crude) as a brown oil. MS (ESI): mass calcd. For C ₅ ILBrNO2S2 254.90, m/z found 256.0 [M+l] ⁺ .

[00538] Step 2: A mixture of 5-bromo-N-methyl-thiophene-2-sulfonamide (1.6 g, 6.25 mmol, 1 eq), phenylmethanethiol (853 mg, 6.87 mmol, 805 μL, 1.1 eq), DIEA (1.61 g, 12.5 mmol, 2.18 mL, 2 eq), Pd(dppf)C12 (114 mg, 156 pmol, 0.025 eq) and (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4- yl)-diphenyl- phosphane (361 mg, 625 pmol, 0.1 eq) in Tol (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 110 °C for 12 hours under N2 atmosphere. LCMS showed 5-bromo-N-methyl-thiophene-2-sulfonamide was consumed and desired mass was detected. The reaction mixture was added to water (20 mL), the aqueous phase was extracted with EtOAc (60 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/ EtOAc = 5/1 to 3/1) to give desired 5-benzylsulfanyl-N-methyl-thiophene-2-sulfonamide (1.5 g, 5.01 mmol, 80.2% yield) as an orange oil. MS (ESI): mass calcd. For C12H13NO2S3. 299.01, m/z found 300.1 [M+l] ⁺ . [00539] Step 3: To a solution of 5-benzylsulfanyl-N-methyl-thiophene-2-sulfonamide (1.5 g, 5.01 mmol, 1 eq) in AcOH (16 mL) and H ₂ O (4 mL) was added NCS (2.01 g, 15.0 mmol, 3 eq). The mixture was stirred at 0 °C for 3 hours. LCMS showed 5-benzylsulfanyl-N-methyl-thiophene-2- sulfonamide was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a crude oil. The residue was purified by pre p-HPLC (column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (10mM NH4HCOs)-ACN]; B%: 35% - 65%, 8 min) to give desired 5 -(methylsulfamoyl) thiophene-2-sulfonyl chloride (39.1 mg, 87.5 pmol, 6.73% yield, 99.7 % purity) as a white solid. MS (ESI): mass calcd. For C5H5CINO4S3 274.91, m/z found 276.0 [M+l] ⁺ .

[00540] Step 4. To a solution of 5 -(methylsulfamoyl) thiophene-2-sulfonyl chloride (300 mg, 1.09 mmol, 1 eq) in Py (5 mL) was added 3 -chloro-2-(4,4-dimethyl-l -piperidyl) aniline (312 mg, 1.31 mmol, 1.2 eq). The mixture was stirred at 15 °C for 3 hours. LCMS showed 5- (methylsulfamoyl) thiophene-2-sulfonyl chloride was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a crude oil. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 3/1). The purified product was dissolved in water (20 mL), and then the solution was lyophilized to give desired N2-[3-chloro-2-(4,4-dimethyl-l- piperidyl) phenyl] -N5-methyl-thiophene-2, 5 -disulfonamide (258 mg, 530 pmol, 48.7% yield, 98.2 % purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.50 (s, 1 H) δ.13 - 7.98 (m, 1 H) 7.59 (s, 2 H) 7.37 - 7.14 (m, 3 H) 3.30 (br s, 2 H) 2.50 (d, J= 5.01 Hz, 3 H) 2.33 (br d, J= 4.77 Hz, 2 H) 1.64 - 1.39 (m, 2 H) 1.38 - 1.22 (m, 2 H) 0.98 (br s, 6 H). HPLC: 98.15% (220 nm), 98.37% (215 nm), 99.17% (254 nm). MS (ESI): mass calcd. For C18H24CIN3O4S3 477.06 m/z found 478.1[M+1] ⁺ .

Example 112: N2-[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] thiophene-2, 5-disulfonamide

[00541] Step 1. To a solution of 5-bromothiophene-2-sulfonyl chloride (2 g, 7.65 mmol, 1 eq) in ammonia (1.95 g, 115 mmol, 1.92 mL, 15 eq). The mixture was stirred at 15 °C for 0.2 hour. TLC (petroleum ether/EtOAc = 3/1) indicated 5-bromothiophene-2-sulfonyl chloride was consumed completely and new spot formed. The reaction mixture was concentrated under reduced pressure to give a residue. The mixture was dissolved in EtOAc (20 mL), and then strried at 20 °C for 5 mins. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired 5-bromothiophene-2-sulfonamide (1.5 g, crude) as a white solid.

[00542] Step 2'. A mixture of 5-bromothiophene-2-sulfonamide (1.5 g, 6.20 mmol, 1 eq), phenylmethanethiol (1.33 g, 10.7 mmol, 1.25 mL, 1.73 eq), DIEA (1.60 g, 12.4 mmol, 2.16 mL, 2 eq), Pd(dppf)C12 (113 mg, 155 pmol, 0.025 eq) and (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4- yl)-diphenyl-phosphane (358 mg, 620 pmol, 0.1 eq) in Tol. (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100 °C for 12 hours under N2 atmosphere. LC-MS showed 5-bromothiophene-2-sulfonamide was consumed completely and desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove Tol. The mixture was cooled at 20 °C and added H ₂ O (100 mL), and extracted with EtOAc (150 mL). The combined organic layers were washed with brine (100 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 1/0 to 2/1) to give desired 5-benzylsulfanylthiophene-2-sulfonamide (1.5 g, 5.26 mmol, 84.8% yield) as a white solid. MS (ESI): mass calcd. For C11H11NO2S3 285.00, m/z found 286.2 [M+l] ⁺ .

[00543] Step 3. To a solution of 5-benzylsulfanylthiophene-2-sulfonamide (1.5 g, 5.26 mmol, 1 eq) in AcOH (16 mL) and H ₂ O (4 mL) was added NCS (2.11 g, 15.8 mmol, 3 eq). The mixture was stirred at 10 °C for 3 hours. LC-MS showed 5-benzylsulfanylthiophene-2-sulfonamide was consumed completely and desired mass was detected. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (400 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 1/0 to 2/1) to give desired 5-sulfamoylthiophene-2-sulfonyl chloride (1 g, 3.82 mmol, 72.7% yield) as a yellow oil.

[00544] Step 4. To a solution of 5-sulfamoylthiophene-2-sulfonyl chloride (200 mg, 764 pmol, 1 eq) in Py (0.5 mL) was added 3-chloro-2-(4, 4-dimethyl-l -piperidyl) aniline (274 mg, 1.15 mmol, 1.5 eq). The mixture was stirred at 15 °C for 1 hour. LC-MS showed 5-sulfamoylthiophene-2-sulfonyl chloride was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 5/1) to give desired N2-[3-chloro-2-(4, 4- dimethyl-1 -piperidyl) phenyl] thiophene-2, 5 -disulfonamide (60.1 mg, 130 pmol, 17.0% yield) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.45 (s, 1 H) δ.03 (s, 2 H) 7.60 - 7.56 (m, 1 H) 7.56 - 7.52 (m, 1 H) 7.34 - 7.25 (m, 2 H) 7.24 - 7.16 (m, 1 H) 3.35 - 3.25 (m, 2 H) 2.36 (br d, J= 8.56 Hz, 2 H) 1.52 (br s, 2 H) 1.35 (br s, 2 H) 0.99 (br s, 6 H). HPLC: 98.26% (220 nm), 98.54% (215 nm), 99.56% (254 nm). MS (ESI): mass calcd. For C17H22CIN3O4S3 463.05 m/z found 463.9 [M+l] ⁺ .

Example 113: N5-[3-chloro-2-(4-ethylpiperazin-l-yl) phenyl] -N2, N2-dimethyl- thiophene-2, 5- disulfonamide

[00545] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (10 g, 57.0 mmol, 1 eq) in DCM (100 mL) was added DIEA (14.7 g, 114 mmol, 19.8 mL, 2 eq) and tert-butyl piperazine-1- carboxylate (11.7 g, 62.7 mmol, 1.1 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed no l-chloro-2-fluoro-3 -nitro-benzene remained. Several new peaks were shown on LC-MS and desired compound was detected. The reaction mixture was diluted with H ₂ O (100 mL) and extracted with EtOAc (900 mL). The combined organic layers were washed with brine (300 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120g SepaFlash® Silica Flash Column, Eluent of 0-10% EtOAc/petroleum ether gradient @ 80 mL/min) to give desired tert-butyl 4-(2-chloro-6-nitro-phenyl) piperazine- 1 -carboxylate (6.1 g, 17.9 mmol, 31.3% yield) as ayellow oil. MS (ESI): mass calcd. For C15H20CIN3O4 341.11, m/z found 242.1 [M-100] ⁺ .

[00546] Step 2: To a solution of tert-butyl 4-(2-chloro-6-nitro-phenyl) piperazine- 1 -carboxylate (6.1 g, 17.9 mmol, 1 eq) in EtOH (50 mL) and H ₂ O (10 mL) was added Fe (9.97 g, 178 mmol, 10 eq) and NH4CI (9.55 g, 178 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed tert-butyl 4-(2-chloro-6-nitro-phenyl) piperazine- 1 -carboxylate was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (300 mL) and extracted with EtOAc (600 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0-35% EtOAc /petroleum ether gradient @ 85 mL/min) to give desired tert-butyl 4-(2-amino-6-chloro-phenyl) piperazine- 1 -carboxylate (4.16 g, 13.3 mmol, 74.8% yield) as a white solid. MS (ESI): mass calcd. For C15H22CIN3O2 311.14, m/z found 256.2 [M-56] ⁺ . [00547] Step 3. To a solution of tert-butyl 4-(2-amino-6-chloro-phenyl) piperazine- 1 -carboxylate (3 g, 9.62 mmol, 1 eq) in Py (10 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (3.07 g, 10.6 mmol, 1.1 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed tert-butyl 4-(2-amino-6-chloro-phenyl) piperazine- 1 -carboxylate was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0-40% EtOAc/petroleum ether gradient @ 85 mL/min) to give desired tert-butyl 4-[2-chloro-6-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonylamino] phenyl] piperazine- 1 -carboxylate (4.4 g, 7.79 mmol, 80.9% yield) as a colourless oil. MS (ESI): mass calcd. For C21H29CIN4O6S3 564.09, m/z found 509.1 [M-56] ⁺ .

[00548] Step 4: A solution of tert-butyl 4-[2-chloro-6-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonylamino] phenyl] piperazine- 1 -carboxylate (4.4 g, 7.79 mmol, 1 eq) in HCl/MeOH (40 mL) was stirred at 20 °C for 12 hours. LC-MS showed tert-butyl 4-[2-chloro-6-[[5-(dimethylsulfamoyl)- 2-thienyl] sulfonylamino] phenyl] piperazine- 1 -carboxylate was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give desired N5 -(3 -chloro-2-piperazin-l-yl-phenyl)-N2, N2-dimethyl-thiophene-2, 5- disulfonamide (3.8 g, crude, HC1) as a white solid. MS (ESI): mass calcd. For C16H21CIN4O4S3 464.04, m/z found 465.1 [M+l] ⁺ .

[00549] Step 5: To a solution of N5-(3-chloro-2-piperazin-l-yl-phenyl)-N2, N2-dimethyl- thiophene-25-disulfonamide (300 mg, 598 pmol, 1 eq, HC1) in MeOH (3 mL) was added TEA (121 mg, 1.20 mmol, 167 μL, 2 eq), the mixture was stirred at 20 °C for 0.1 hour, acetaldehyde (31.6 mg, 718 pmol, 40.3 μL, 1.2 eq) and AcOH (108 mg, 1.79 mmol, 103 μL, 3 eq) was added to the mixture at 20 °C and stirred for 0.5 hours. NaBFFCN (75.2 mg, 1.20 mmol, 2 eq) was added to the mixture at 0 °C. The mixture was stirred at 0-20 °C for 3 hours. LC-MS showed N5-(3-chloro-2-piperazin-l- yl-phenyl)-N2, N2-dimethyl-thiophene-25-disulfonamide was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-WY,C (neutral condition: column: Phenomenex Gemini-NX C18 75*30mm*3pm; mobile phase: [water (10mM NH ₄ HCO ₃ )-ACN];B%: 25%-55%,10 min) to give desired N5-[3-chloro-2-(4-ethylpiperazin-l-yl) phenyl] -N2, N2-dimethyl-thiophene-2, 5-disulfonamide (54 mg, 110 pmol, 18.3% yield, 100% purity) as a white solid ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 7.47 (d, J= 3.9 Hz, 1H), 7.34 (br d, J= 3.8 Hz, 1H), 7.16 (br d, J= 8.0 Hz, 1H), 6.86 (br t, J= 8.0 Hz, 1H), 6.78 (br d, J= 7.6 Hz, 1H), 3.63 (br s, 2H), 3.24 (br s, 2H), 3.01 - 2.82 (m, 6H), 2.63 (s, 6H), 1.19 (br t, J= 7.2 Hz, 3H). HPLC: 100.00% (220 nm), 100.00% (215 nm), 100.00% (254 nm). MS (ESI): mass calcd. For C18H25CIN4O4S3 492.07 m/z found 493.1 [M+l] ⁺ .

Example 114: N5-[3-chloro-2-(4-isopropylpiperazin-l-yl) phenyl] -N2, N2-dimethyl-thiophene- 2, 5-disulfonamide

[00550] Step 1. To a solution of acetone (174 mg, 2.99 mmol, 220 μL, 5 eq) and N5-(3-chloro-2- piperazin-l-yl-phenyl)-N2, N2-dimethyl-thiophene-2, 5-disulfonamide (300 mg, 598 pmol, 1 eq, HC1) in MeOH (1 mL) was added TEA (60.5 mg, 598 pmol, 83 μL, 1 eq) until pH~8. The mixture was stirred at 20 °C for 0.1 hour. CH3COOH (108 mg, 1.79 mmol, 103 μL, 3 eq) was added to the mixture until pH~5. The mixture was stirred at 20 °C for 0.5 hour. NaBHsCN (188 mg, 2.99 mmol, 5 eq) was added to the mixture at 0 °C. The mixture was stirred at 20 °C for 1 hour. LC-MS showed N5-(3-chloro-2-piperazin-l-yl-phenyl)-N2, N2-dimethyl-thiophene-2, 5-disulfonamide remained and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WY,C (column: Phenomenex Gemini- NX C18 75*30mm*3um; mobile phase: [water (0.05% NH3.H ₂ O + 10 mM NH4HCO ₃ )-ACN]; B%: 30%-60%, 8min) to give desired N5-[3-chloro-2-(4-isopropylpiperazin-l-yl) phenyl]-N2, N2- dimethyl-thiophene-2, 5-disulfonamide as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 7.47 (d, J = 3.75 Hz, 1 H) 7.33 (br d, J= 3.63 Hz, 1 H) 7.17 (br d, J= 7.88 Hz, 1 H) 6.85 (br t, J= 8.00 Hz, 1 H) 6.75 (br d, J= 7.00 Hz, 1 H) 3.81 - 3.62 (m, 2 H) 3.00 - 2.83 (m, 4 H) 2.77 (s, 1 H) 2.68 (br d, J = 1.63 Hz, 2 H) 2.64 (s, 6 H)1.24 (br d, J= 6.38 Hz, 6 H). HPLC: 94.94% (220 nm), 95.11% (215 nm), 95.10% (254 nm). MS (ESI): mass calcd. For C19H27CIN4O4S3 506.09 m/z found 507.2 [M+H] ⁺ .

Example 115: N5-[3-chloro-2- [cyclohexyl (methyl) amino] phenyl] -N2, N2-dimethyl-thiophene- 2, 5-disulfonamide

[00551] Step 1. To a solution of N-methylcyclohexanamine (1 g, 8.83 mmol, 1.17 mL, 1 eq) in dioxane (15 mL) was added K ₂ CO ₃ (3.66 g, 26.5 mmol, 3 eq) and 1 -chloro-2-fluoro-3 -nitro-benzene (1.71 g, 9.72 mmol, 1.1 eq). The mixture was stirred at 100 °C for 12 hours. LCMS showed N- methylcyclohexanamine was consumed and desired mass was detected. The crude was added H2O (15 mL), and extracted with EtOAc (45 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 5/1 to 4/1) to give desired 2-chloro-N- cyclohexyl-N-methyl-6-nitro-aniline (0.5 g, 1.86 mmol, 21.1% yield) as a brown oil. MS (ESI): mass calcd. For C13H17CIN2O2 268.10 m/z found 269.2 [M+H] ⁺ .

[00552] Step 2: To a solution of 2-chloro-N-cyclohexyl-N-methyl-6-nitro-aniline (0.5 g, 1.86 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (0.5 mL) was added Fe (520 mg, 9.30 mmol, 5 eq) and NH4CI (995 mg, 18.6 mmol, 10 eq). The mixture was stirred at 90 °C for 12 hours. LCMS showed 2- chloro-N-cyclohexyl-N-methyl-6-nitro-aniline was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude oil. The crude was added H ₂ O (10 mL), and extracted with EtOAc (30 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 1/0 to 10/1) to give desired 3-chloro-N2-cyclohexyl-N2-methyl-benzene-l, 2-diamine (190 mg, 796 pmol, 42.8% yield) as a brown oil. MS (ESI): mass calcd. For C13H19CIN2 238.12 m/z found 239.2 [M+H] ⁺ .

[00553] Step 3. To a solution of 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (254 mg, 875 pmol, 1.1 eq) in Py (2 mL) was added 3-chloro-N2-cyclohexyl-N2-methyl-benzene-l, 2-diamine (190 mg, 796 pmol, 1 eq). The mixture was stirred at 15 °C for 12 hours. LCMS showed 5- (dimethylsulfamoyl) thiophene-2-sulfonyl chloride was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a crude oil. The residue was purified by prep-WL,C (column: Phenomenex Luna C18 200*40mm*10um; mobile phase: [water (0.2%FA)-ACN]; B%: 50%-90%, 8min) to give desired N5- [3 -chloro-2- [cyclohexyl (methyl) amino] phenyl]-N2, N2-dimethyl-thiophene-2, 5-disulfonamide (84.9 mg, 173 pmol, 21.7% yield, 97.0 % purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 9.46 - 9.39 (m, 1 H) 7.83 - 7.76 (m, 1 H) 7.69 - 7.62 (m, 1 H) 7.52 - 7.45 (m, 1 H) 7.29 - 7.13 (m, 2 H) 3.00 (br s, 1 H) 2.63 (s, 6 H) 2.50 (s, 3 H) 2.05 (s, 1 H) 1.68 - 1.78 (m, 1 H) 1.68 (br s, 1 H) 1.47 (br d, J= 9.65 Hz, 2 H) 1.13 (br s, 3 H) 1.01 (br s, 2 H) 0.46 - 0.25 (m, 1 H). HPLC: 96.97% (220 nm), 96.34% (215 nm), 98.26% (254 nm). MS (ESI): mass calcd. For C19H26CIN3O4S3 491.08 m/z found 492.3 [M+H] ⁺ .

Example 116: N5-[3-chloro-2-[4-(2-methylpropanoyl) piperazin- 1-yl] phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00554] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (10 g, 57.0 mmol, 1 eq) in DCM (100 mL) was added DIEA (14.7 g, 114 mmol, 19.8 mL, 2 eq) and tert-butyl piperazine-1- carboxylate (11.7 g, 62.7 mmol, 1.1 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS 1- chloro-2-fluoro-3 -nitro-benzene showed no remained. Several new peaks were shown on LC-MS and desired compound was detected. The reaction mixture was diluted with H ₂ O (100) mL and extracted with EtOAc (900 mL). The combined organic layers were washed with brine (300 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120g SepaFlash® Silica Flash Column, Eluent of 0-10% EtOAc /petroleum ether gradient @ 80 mL/min) to give desired tert-butyl 4-(2-chloro-6-nitro-phenyl) piperazine- 1 -carboxylate (6.1 g, 17.9 mmol, 31.3% yield) as ayellow oil. MS (ESI): mass calcd. For C15H20CIN3O4 341.11 m/z found 242.1 [M-100] ⁺ . [00555] Step 2\ To a solution of tert-butyl 4-(2-chloro-6-nitro-phenyl) piperazine- 1 -carboxylate (6.1 g, 17.9 mmol, 1 eq) in EtOH (50 mL) and H ₂ O (10 mL) was added Fe (9.97 g, 178 mmol, 10 eq) and NH4CI (9.55 g, 178 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed tert-butyl 4-(2-chloro-6-nitro-phenyl) piperazine- 1 -carboxylate was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (300 mL) and extracted with EtOAc (600 mL). The combined organic layers were washed with brine 150 (mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0-35% EtOAc/petroleum ether gradient @ 85 mL/min) to give desired tert-butyl 4-(2-amino-6-chloro-phenyl) piperazine- 1 -carboxylate (4.16 g, 13.3 mmol, 74.8% yield) as a white solid. MS (ESI): mass calcd. For C15H22CIN3O2 311.14 m/z found 256.2 [M-56] ⁺ .

[00556] Step 3: To a solution of tert-butyl 4-(2-amino-6-chloro-phenyl) piperazine- 1 -carboxylate (3 g, 9.62 mmol, 1 eq) in pyridine (10 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (3.07 g, 10.6 mmol, 1.1 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed tert-butyl 4-(2-amino-6-chloro-phenyl) piperazine- 1 -carboxylate was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0-40% EtOAc/petroleum ether gradient @ 85 mL/min) to give desired tert-butyl 4-[2-chloro-6-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonylamino] phenyl] piperazine- 1 -carboxylate (4.4 g, 7.79 mmol, 80.9% yield) as a colorless oil. MS (ESI): mass calcd. For C21H29CIN4O6S3 564.09 m/z found 506.1 [M-56] ⁺ .

[00557] Step 4: A solution of tert-butyl 4-[2-chloro-6-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonylamino] phenyl] piperazine- 1 -carboxylate (4.4 g, 7.79 mmol, 1 eq) in HCl/MeOH (40 mL) was stirred at 20 °C for 12 hours. LC-MS showed tert-butyl 4-[2-chloro-6-[[5-(dimethylsulfamoyl)- 2-thienyl] sulfonylamino] phenyl] piperazine- 1 -carboxylate was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give desired N5 -(3 -chloro-2-piperazin-l-yl-phenyl)-N2, N2-dimethyl-thiophene-2, 5- disulfonamide (3.8 g, crude, HC1) as a white solid. MS (ESI): mass calcd. For C16H21CIN4O4S3 464.04 m/z found 465.1 [M+l] ⁺ .

[00558] Step 5: To a solution of N5-(3-chloro-2-piperazin-l-yl-phenyl)-N2, N2-dimethyl- thiophene-2, 5 -disulfonamide (200 mg, 399 pmol, 1 eq, HC1) and 2-methylpropanoyl chloride (46.7 mg, 439 pmol, 45.8 μL, 1.1 eq) in DCM (2 mL) was added TEA (202 mg, 1.99 mmol, 278 μL, 5 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed N5-(3-chloro-2-piperazin-l-yl- phenyl)-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by pre p-HPLC (neutral condition: column: Waters Xbridge BEH C18 100*25mm*5um; mobile phase: [water (10mM NH4HCOs)-ACN]; B%: 30%- 65%, 10 min) to give desired N5-[3-chloro-2-[4-(2-methylpropanoyl) piperazin-l-yl] phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (58.1 mg, 105 pmol, 26.4% yield, 96.9% purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.93 (s, 1H), 7.69 (q, J= 4.0 Hz, 2H), 7.35 - 7.29 (m, 1H), 7.24 - 7.19 (m, 2H), 3.79 (br s, 1H), 3.63 (br s, 2H), 3.31 - 3.07 (m, 3H), 2.95 - 2.85 (m, 1H), 2.68 (s, 6H), 2.60 (br s, 1H), 2.42 (br s, 1H), 1.03 (s, 3H), 1.01 (s, 3H). HPLC: 96.94% (220 nm), 97.32% (215 nm), 97.79% (254 nm). MS (ESI): mass calcd. For C20H27CIN4O5S3 534.08 m/z found 535.2 [M+l] ⁺ .

Example 117: N5-[2-(2, 6-dimethyl-l-piperidyl) phenyl]-N2, N2-dimethyl-thiophene-2, 5- disulfonamide

[00559] Step 1. To a solution of glutaric acid (5.00 g, 37.8 mmol, 1 eq) in SOCL (50 mL) was added DMF (0.5 mL). The mixture was stirred at 90 °C for 5 hours. TLC (petroleum ether/EtOAc = 3/1) indicated glutaric acid was consumed completely and one new spot formed. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired pentanedioyl dichloride (6 g, crude) as a brown oil.

[00560] Step 2: To a solution of N-methoxymethanamine (6.51 g, 107 mmol, 3 eq) in DCM (100 mL) was added TEA (21.6 g, 2137 mmol, 30.0 mL, 6 eq) at 0 °C. After addition, and then pentanedioyl di chloride (6 g, 35.5 mmol, 4.55 mL, 1 eq) was added dropwise at 0 °C. The resulting mixture was stirred at 15 °C for 0.5 hours. TLC (petroleum ether/EtOAc = 3/1) indicated N- methoxymethanamine was consumed completely and one new spot formed. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired N, N'- dimethoxy-N, N'-dimethyl-pentanediamide (7 g, crude) as a brown oil.

[00561] Step 3. To a solution of N, N'-dimethoxy-N, N'-dimethyl-pentanediamide (7 g, 32.07 mmol, 1 eq) in THF (70 mL) was added bromo (methyl) magnesium (3 M, 32.1 mL, 3 eq) at 0 °C. The resulting mixture was stirred at 0 °C for 1 hours. TLC (petroleum ether/EtOAc = 3/1) indicated N, N'-dimethoxy-N, N'-dimethyl-pentanediamide was consumed completely and one new spot formed. The crude was added H ₂ O (100 mL), and extracted with EtOAc (300 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/ EtOAc = 6/1 to 5/1) to give desired heptane-2, 6-dione (2.1 g, 16.4 mmol, 51.1% yield) as ayellow oil.

[00562] Step 4. To a solution of heptane-2, 6-dione (185 mg, 1.44 mmol, 1.5 eq) andtert-butyl N- (2-aminophenyl) carbamate (200 mg, 960 pmol, 1 eq) in MeOH (20 mL) was AcOH (57.7 mg, 960 pmol, 54.9 μL, 1 eq) added at 0 °C. After addition, the mixture was stirred at this temperature for 1 hours, and then NaBHsCN (302 mg, 4.80 mmol, 5 eq) was added dropwise at 0 °C. The resulting mixture was stirred at 20 °C for 12 hours. LCMS showed heptane-2, 6-dione was consumed and desired mass was detected. The crude was added H ₂ O (10 mL), and extracted with EtOAc (30 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether/EtOAc = 4/1) to give desired tert-butyl N-[2-(2, 6-dimethyl-l -piperidyl) phenyl] carbamate (155 mg, 509 pmol, 53.0% yield) as a brown oil. MS (ESI): mass calcd. For C18H28N2O2 304.22m/z found 305.3 [M+l] ⁺ .

[00563] Step 5: To a solution of tert-butyl N-[2-(2, 6-dimethyl-l -piperidyl) phenyl] carbamate (150 mg, 493 pmol, 1 eq) in HCl/MeOH (2 mL).The mixture was stirred at 15 °C for 0.5 hour. TLC (petroleum ether/EtOAc = 3/1) indicated tert-butyl N-[2-(2, 6-dimethyl-l -piperidyl) phenyl] carbamate was consumed completely and one new spot formed. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired 2-(2, 6-dimethyl-l -piperidyl) aniline (110 mg, crude) as a brown oil.

[00564] Step 6: To a solution of 2-(2, 6-dimethyl-l -piperidyl) aniline (110 mg, 538 pmol, 1 eq) in Py (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (172mg, 592pmol, 1.1 eq). The mixture was stirred at 15 °C for 12 hours. LCMS showed 2-(2, 6-dimethyl-l -piperidyl) aniline was consumed and desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove solvent to give a residue (220 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C19H27N3O4S3 457.12 m/z found 458.3 [M+l] ⁺ .

[00565] Step 7: N5-[2-(2, 6-dimethyl-l -piperidyl) phenyl]-N2, N2-dimethyl-thiophene-2, 5- disulfonamide (220 mg, crude) was purified by prep-HPLC (column: Phenomenex Luna Cl 8 200*40mm*10um; mobile phase: [water (0.2%FA)-ACN]; B%: 40%-80%, 8 min). The purified product was dissolved in water (20 mL), and then the solution was lyophilized to give desired N5-[2- (2, 6-dimethyl-l -piperidyl) phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (12.7 mg, 27.8 pmol, 100.00 % purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 7.85 - 7.77 (m, 1 H) 7.65 - 7.56 (m, 2 H) 7.39 - 7.31 (m, 1 H) 7.27 - 7.20 (m, 1 H) 7.15 - 7.05 (m, 1 H) 2.85 - 2.37 (m, 2 H) 2.68 - 2.62 (m, 1 H) 2.58 (s, 6 H) 2.33 - 2.28 (m, 1 H) 1.69 (br s, 2 H) 1.46 - 1.39 (m, 2 H) 0.39 (d, J = 6.36 Hz, 6 H). HPLC: 100.00% (220 nm), 99.77% (215 nm), 100% (254 nm). MS (ESI): mass calcd. For C19H27N3O4S3 457.12 m/z found 458.3 [M+l] ⁺ .

Example 118: N5-[2-(2, 6-dimethyl-l-piperidyl) phenyl] -N2, N2-dimethyl- thiophene-2, 5- disulfonamide

[00566] Step 1. N5-[2-(2, 6-dimethyl-l-piperidyl) phenyl]-N2, N2-dimethyl-thiophene-2, 5- disulfonamide (220 mg, crude) was purified by prep-WY,C (column: Phenomenex Luna Cl 8 200*40mm*10um; mobile phase: [water (0.2%FA)-ACN]; B%: 40%-80%, 8 min). The purified product was dissolved in water (20 mL), and then the solution was lyophilized to give desired N5-[2- (2, 6-dimethyl-l-piperidyl) phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (6.9 mg, 15.1 pmol, 99.6 % purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 7.75 - 7.69 (m, 1 H) 7.64 - 7.60 (m, 1 H). 7.64 - 7.60 (m, 1 H) 7.52 - 7.46 (m, 1 H) 7.18 - 7.12 (m, 1 H) 7.11 - 7.06 (m, 2 H) 2.67 - 2.63 (m, 1 H) 2.33 - 2.28 (m, 1 H) 2.61 (s, 6 H) 1.82 (br s, 1 H) 1.68 - 1.28 (m, 5 H) 0.78 (br d, J = 6.58 Hz, 3 H) 0.52 (br d, J= 5.92 Hz, 3 H). HPLC: 99.64% (220 nm), 99.62% (215 nm), 100% (254 nm). MS (ESI): mass calcd. For C19H27N3O4S3 457.12 m/z found 458.3 [M+l] ⁺ . Example 119: N5-[2-(8-azabicyclo [3.2.1] octan-8-yl)-3-chloro-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00567] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (300 mg, 1.71 mmol, 1 eq) and 8-azabicyclo [3.2.1] octane (190 mg, 1.71 mmol, 1 eq) in dioxane (5 mL) was added K ₂ CO ₃ (472 mg, 3.42 mmol, 2 eq) and DIEA (442 mg, 3.42 mmol, 595 μL, 2 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (450 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, Petroleum ether/EtOAc = 10/1) to give desired 8-(2- chloro-6-nitro-phenyl)-8-azabicyclo [3.2.1] octane (160 mg, 600 pmol, 35.1% yield) as ayellow oil. MS (ESI): mass calcd. For C13H15CIN2O2 266.08 m/z found 267.3 [M+l] ⁺ .

[00568] Step 2.' To a solution of 8-(2-chloro-6-nitro-phenyl)-8-azabicyclo [3.2.1] octane (160 mg, 600 pmol, 1 eq) in H ₂ O (0.5 mL) and EtOH (2 mL) was added Fe (26 mg, 4.80 mmol, 8 eq) and NH4CI (481 mg, 9.00 mmol, 15 eq). The mixture was stirred at 80 °C for 2 hours. LC-MS showed 8-(2-chloro-6-nitro-phenyl)-8-azabicyclo [3.2.1] octane was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was added H ₂ O (20mL) and extracted with EtOAc (60 mL). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether/EtOAc = 5/1) to give desired 2-(8-azabicyclo [3.2.1] octan-8-yl)-3 -chloro-aniline (120 mg, 507 pmol, 84.5% yield) as a brown oil. MS (ESI): mass calcd. For C13H17CIN2 236.11 m/z found 237.3 [M+l] ⁺ .

[00569] Step 3. To a solution of 2-(8-azabicyclo [3.2.1] octan-8-yl)-3 -chloro-aniline (120 mg, 507 pmol, 1 eq) in Py (1 mL) was added 5-(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (220 mg, 760 pmol, 1.5 eq). The mixture was stirred at 15 °C for 12 hours. LC-MS showed 2-(8- azabicyclo [3.2.1] octan-8-yl)-3 -chloro-aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WLC (neutral condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (10mM NEEHCO3)-ACN]; B%: 50%-80%,10 min) to give desired N5-[2-(8-azabicyclo [3.2.1] octan-8-yl)-3-chloro-phenyl]-N2, N2- dimethyl-thiophene-2, 5-disulfonamide (24.3 mg, 46.9 pmol, 9.25% yield, 94.6% purity) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.89 (br s, 1 H)7.70 (br dd, J= 19.20, 3.69 Hz, 2 H) 7.29 (br d, J= 7.63 Hz, 1 H) 6.73 (t, J= 7.88 Hz, 1 H) 6.49 (br d, J= 7.75 Hz, 1 H) 3.93 (br s, 2 H) 2.72 (s, 6 H) 2.21 - 2.11 (m, 2 H) 1.99 (br d, J= 7.50 Hz, 2 H) 1.73 (br d, J= 7.00 Hz, 3 H) 1.62 - 1.35 (m, 1 H) 1.46 (br d, J= 11.38 Hz, 2 H). HPLC: 94.56% (220 nm), 94.36% (215 nm), 98.58% (254 nm). MS (ESI): mass calcd. For C19H24CIN3O4S3 489.06 m/z found 489.9 [M+l] ⁺ .

Example 120: N5-[3-chloro-2-(2, 4, 4-trimethyl-l-piperidyl) phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

120

[00570] Step 1. To a solution of 2, 4, 4-trimethylcyclopentanone (10 g, 79.2 mmol, 224 μL, 1 eq) in HCOOH (100 mL) was added amino hydrogen sulfate (13.4 g, 119 mmol, 1.5 eq). The mixture was stirred at 20 °C for 0.5 hour. The reaction mixture was heated at 100 °C for 12 hours. LC-MS showed 2, 4, 4-trimethylcyclopentanone was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove HCOOH. The reaction mixture was partitioned between NaOH (4N, 100 mL) and EtOAc (300 mL). The organic phase was separated, washed with brine (100 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by prep- HPLC (column: Phenomenex Titank C18 Bulk 250*100mm 10 u; mobile phase: [water (10 mM NILHCO3)-ACN]; B%: 15%-35%, 20 min) to give desired 4, 4, 6-trimethylpiperidin-2-one (2.8 g, 19.8 mmol, 28.0% yield) as a white solid. MS (ESI): mass calcd. For CsHisNO 141.12 m/z found 142.2 [M+l] ⁺ . [00571] Step 2\ To a solution of 4, 4, 6-trimethylpiperidin-2-one (1 g, 7.08 mmol, 1 eq) in THF (10 mL) was added LAH (806 mg, 21.2 mmol, 3 eq) at 0 °C. After addition, the mixture was stirred at 70 °C for 12 hours. TLC (petroleum ether/EtOAc = 0/1) indicated 4, 4, 6-trimethylpiperidin-2-one was consumed completely and one new spot formed. The reaction mixture was quenched by addition H ₂ O (0.81 mL), NaOH (0.81 mL, 15%) and H ₂ O (2.43 mL) at 0 °C, the reaction mixture was filtered, and the filtrate was added HCl/EtOAc, and was concentrated under reduced pressure to give desired 2, 4, 4-trimethylpiperidine (1 g, crude, HC1) as a yellow solid.

[00572] Step 3: A mixture of 1 -chloro-2-fluoro-3 -nitro-benzene (579 mg, 3.30 mmol, 37.3 μL, 0.9 eq), 2, 4, 4-trimethylpiperidine (600 mg, 3.67 mmol, 1 eq, HC1) , K ₂ CO ₃ (1.01 g, 7.33 mmol, 2 eq) and DIEA (947 mg, 7.33 mmol, 1.28 mL, 2 eq) in dioxane (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 140 °C for 12 hours under N2 atmosphere. TLC (petroleum ether/EtOAc = 10/1) indicated -70% of l-chloro-2-fluoro-3 -nitro-benzene remained, and two new spot with lower polarity was detected. The reaction mixture was partitioned between water (100 mL) and EtOAc (300 mL). The organic phase was separated, washed brine (200 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-0% EtOAc/petroleum ether gradient @ 80 mL/min) to give desired 1 -(2-chloro-6-nitro-phenyl)- 2, 4, 4-trimethyl-piperidine (0.4 g, 1.41 mmol, 12.9% yield) yellow oil.

[00573] Step 4. To a solution of l-(2-chloro-6-nitro-phenyl)-2, 4, 4-trimethyl-piperidine (50 mg, 177 pmol, 1 eq) in EtOH (1 mL) and H ₂ O (0.2 mL) was added Fe (49.4 mg, 884 pmol, 5 eq) and AcOH (42.5 mg, 707 pmol, 4 eq). The mixture was stirred at 80 °C for 0.5 hour. TLC (petroleum ether/EtOAc = 10/1) indicated l-(2-chloro-6-nitro-phenyl)-2, 4, 4-trimethyl-piperidine was consumed completely and one new spot formed. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give desired 3-chloro-2-(2, 4, 4-trimethyl-l -piperidyl) aniline (45 mg, crude) yellow oil.

[00574] Step 5: A mixture of 3-chloro-2-(2, 4, 4-trimethyl-l -piperidyl)aniline (45 mg, 178 pmol, 1 eq) and 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (56.7 mg, 196 pmol, 1.1 eq) in Py (1 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 0.5 hour under N2 atmosphere. LC-MS showed a little of 3-chloro-2-(2, 4, 4-trimethyl-l -piperidyl) aniline remained. Several new peaks were shown on LC-MS and desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove Py. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 3/1) to give desired N5-[3-chloro-2-(2, 4, 4- trimethyl-1 -piperidyl) phenyl]-N2, N2-dimethyl-thiophene-2, 5-disulfonamide (15.2 mg, 29.9 pmol, 16.8% yield, 99.6% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.27 (s, 1H), 7.81 (d, J= 4.0 Hz, 1H), 7.67 (d, J= 4.0 Hz, 1H), 7.56 (d, J = 7.7 Hz, 1H), 7.31 (t, J= 8.2 Hz, 1H), 7.22 (d, J= 7.9 Hz, 1H), 3.72 - 3.63 (m, 1H), 3.34 (br s, 1H), 3.31 - 3.27 (m, 1H), 2.63 (s, 6H), 1.75 (br d, J= 11.6 Hz, 1H), 1.53 - 1.44 (m, 1H), 1.35 (br s, 1H), 1.23 (br d, J= 12.5 Hz, 1H), 0.97 (s, 6H), 0.48 (d, J= 6.2 Hz, 3H). HPLC: 99.51% (220 nm), 99.49% (215 nm), 99.43% (254 nm). MS (ESI): mass calcd. For C20H28CIN3O4S3 505.09 m/z found 506.1 [M+l] ⁺ .

Example 121: N5-[2-(4-tert-butylpiperazin-l-yl)-3-chloro-phenyl]-N2, N2-dimethyl-thiophene- 2, 5-disulfonamide

[00575] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (400 mg, 2.28 mmol, 1 eq) in dioxane (4 mL) was added K ₂ CO ₃ (630mg, 4.56 mmol, 2 eq) and 1 -tert-butylpiperazine (486 mg, 3.42 mmol, 1.5 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed l-chloro-2- fluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiCh, petroleum ether/EtOAc = 1/0 to 1/1) to give desired 1 -tert-butyl-4-(2- chloro-6-nitro-phenyl) piperazine (600 mg, 2.01 mmol, 88.4% yield) as a yellow oil. MS (ESI): mass calcd. For C14H20CIN3O2 297.12 m/z found 298.3 [M+l] ⁺ .

[00576] Step 2: To a solution of l-tert-butyl-4-(2-chloro-6-nitro-phenyl) piperazine (400 mg, 1.34 mmol, 1 eq) in H ₂ O (0.4 mL) and EtOH (4 mL) was added Fe (375 mg, 6.72 mmol, 5 eq), CH3COOH (323 mg, 5.37 mmol, 307 μL, 4 eq) and NH4CI (719mg, 13.4 mmol, 10 eq). The mixture was stirred at 80 °C for 12 hours. LC-MS showed l-tert-butyl-4-(2-chloro-6-nitro-phenyl) piperazine was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The reaction mixture was added to water (20 mL), the aqueous phase was extracted with EtOAc (30 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether: EtOAc = 0/1) to give desired 2-(4-tert-butylpiperazin-l-yl)-3- chloro-aniline (320 mg, 1.19 mmol, 89.0% yield) as ayellow oil. MS (ESI): mass calcd. For C14H22CIN3 267.15 m/z found 268.3 [M+l] ⁺ .

[00577] Step 3. To a solution of 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (243 mg, 840 pmol, 1.5 eq) in Py (1 mL) was added 2-(4-tert-butylpiperazin-l-yl)-3 -chloro-aniline (150 mg, 560 pmol, 1 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed 5- (dimethylsulfamoyl) thiophene-2-sulfonyl chloride was consumed completely and desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by prep-HPLC (neutral condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (10mM NH ₄ HCO ₃ )-ACN]; B%: 35%-65%,10 min ) to give desired N5-[2-(4-tert- butylpiperazin-l-yl)-3 -chloro-phenyl] -N2, N2-dimethyl-thiophene-2, 5-disulfonamide (21.4 mg, 40.1 pmol, 7.15% yield, 97.6% purity) as an off-white solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 7.52 (d, J = 3.8 Hz, 1H), 7.38 (br d, J= 3.0 Hz, 1H), 7.22 (br d, J= 8.1 Hz, 1H), 6.94 - 6.84 (m, 1H), 6.83 - 6.73 (m, 1H), 3.99 - 3.77 (m, 2H), 3.55 (br s, 2H), 3.34 - 3.25 (m, 2H), 2.96 (br d, J= 3.1 Hz, 3H), 2.70 (s, 6H), 1.38 (br s, 9H). HPLC: 97.59% (220 nm), 97.88% (215 nm), 99.26% (254 nm). MS (ESI): mass calcd. For C20H29CIN4O4S3 520.10 m/z found 520.9 [M+l] ⁺ .

Example 122: N5-[2-(2-azabicyclo [2.2.1] heptan-2-yl)-3-chloro-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00578] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (350 mg, 1.99 mmol, 1 eq) in 1,4-dioxane (5 mL) was added 2-azabicyclo [2.2.1] heptane (213 mg, 2.19 mmol, 1.1 eq), K ₂ CO ₃ (551 mg, 3.99 mmol, 2 eq) and DIEA (515 mg, 3.99 mmol, 695 μL, 2 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (5 mL) and extracted with EtOAc (60 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 24 g SepaFlash® Silica Flash Column, Eluent of 0-10% EtOAc/petroleum ether gradient @ 35 mL/min) to give desired 2-(2-chloro-6-nitro-phenyl)-2-azabicyclo [2.2.1] heptane (490 mg, 1.94 mmol, 97.3% yield) as a yellow oil. MS (ESI): mass calcd. For C12H13CIN2O2 252.07 m/z found 253.2 [M+l] ⁺ .

[00579] Step 2\ To a solution of 2-(2-chloro-6-nitro-phenyl)-2-azabicyclo [2.2.1] heptanes (490 mg, 1.94 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (1 mL) was added Fe (1.08 g, 19.4 mmol, 10 eq) and NH4CI (1.04 g, 19.4 mmol, 10 eq). The mixture was stirred at 80 °C for 3 hours. LC-MS showed 2- (2-chloro-6-nitro-phenyl)-2-azabicyclo [2.2.1] heptanes was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (5 mL) and extracted with EtOAc (30 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-20% EtOAc /petroleum ether gradient @ 30 mL/min) to give desired 2-(2-azabicyclo [2.2.1] heptan-2-yl)-3 -chloro-aniline (240 mg, 1.08 mmol, 55.6% yield) as a yellow oil. MS (ESI): mass calcd. For C12H15CIN2 222.09 m/z found 223.2 [M+l] ⁺ .

[00580] Step 3. To a solution of 2-(2-azabicyclo [2.2.1] heptan-2-yl)-3 -chloro-aniline (240 mg, 1.08 mmol, 1 eq) in pyridine (3 mL) was added 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (375 mg, 1.29 mmol, 1.2 eq). The mixture was stirred at 20 °C for 2 hours. LC-MS showed 2-(2- azabicyclo [2.2.1] heptan-2-yl)-3 -chloro-aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition: column: Phenomenex luna Cl 8 100*40mm*5 um; mobile phase: [water (0.1%TFA)-ACN]; B%: 70%-95%, 8 min) to give desired N5-[2-(2-azabicyclo [2.2.1] heptan-2-yl)-3 -chloro-phenyl] -N2, N2-dimethyl-thiophene-2, 5- disulfonamide (64.5 mg, 135 pmol, 12.5% yield, 99.6% purity) as a red solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 9.68 (br s, 1H), 7.71 (br s, 2H), 7.28 (br s, 1H), 7.10 - 6.82 (m, 2H), 3.65 (br s, 1H), 3.09 (br s, 1H), 2.71 (br s, 6H), 2.57 (br s, 2H), 2.03 (br s, 1H), 1.70 - 1.48 (m, 4H), 1.36 (br s, 1H). HPLC: 99.64% (220 nm), 99.41% (215 nm), 99.36% (254 nm). MS (ESI): mass calcd. For C18H22CIN3O4S3 475.05 m/z found 476.2 [M+l] ⁺ .

Example 123: N5-[3-chloro-2-(2-oxo-l-piperidyl) henyl]-N2, 2-dimethyl- thiophene-2, 5- disulfonamide [00581] Step 1. 2-chloro-6-nitro-aniline (2 g, 11.6 mmol, 1 eq) was dissolved in THF (20 mL) was added 5-bromopentanoyl chloride (2.31 g, 11.6 mmol, 1.55 mL, 1 eq) and TEA (2.35 g, 23.2 mmol, 3.23 mL, 2 eq) at 0 °C under N2. The mixture was stirred at 20 °C for 1 hour. Potassium tert- butoxide (2.60 g, 23.2 mmol, 2 eq) was added to the mixture at 0 °C. The mixture was stirred at 20 °C for 4 hours. TLC (petroleum ether/EtOAc = 1/1, Rf = 0.70) showed 2-chloro-6-nitro-aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water (100 mL) and extracted with methylene dichloride (600 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 1/0 to 2/1) to give desired l-(2-chloro-6-nitro- phenyl) piperidin-2-one (270 mg, 1.06 mmol, 9.15% yield) as a faint yellow oil.

[00582] Step 2. To a solution of l-(2-chloro-6-nitro-phenyl) piperidin-2-one (120 mg, 471 pmol, 1 eq) in H ₂ O (0.3 mL) and EtOH (1.5 mL) was added Fe (21 Img, 3.77 mmol, 8 eq) and NH4CI (378 mg, 7.07 mmol, 15 eq). The mixture was stirred at 80 °C for 1 hour. LC-MS showed l-(2-chloro-6- nitro-phenyl) piperidin-2-one was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was added H ₂ O (20mL) and extracted with EtOAc (60 mL) The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, EtOAc/Methanol = 10/1) to give desired l-(2-amino-6-chloro-phenyl) piperidin-2-one (45 mg, 200 pmol, 42.5% yield) as a white solid. MS (ESI): mass calcd. For C11H13CIN2O 224.07 m/z found 225.1 [M+l] ⁺ .

[00583] Step 3. To a solution of l-(2-amino-6-chloro-phenyl) piperidin-2-one (100 mg, 445 pmol, 1 eq) in Py (1 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (193 mg, 668 mol, 1.5 eq). The mixture was stirred at 15 °C for 24 hours. LC-MS showed l-(2-amino-6- chloro-phenyl) piperidin-2-one was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WY,C (neutral condition; column: Phenomenex Gemini- NX 80*40mm*3um; mobile phase: [water 10mM NEEHCO3)-ACN];B%: 8%-38%,8 min) to give desired N5- [3 -chloro-2-(2-oxo-l -piperidyl) henyl]-N2, 2-dimethyl-thiophene-2, 5 -disulfonamide (23.3 mg, 46.6 pmol, 10.5% yield, 95.7% purity) as a faint yellow solid. ¹ H NMR (400 MHz, DMSO-d6 ) δ 10.53 (br s, 1 H) 7.65 (br s, 2 H) 7.52 - 7.27 (m, 3 H) 3.19 - 3.06 (m, 1 H) 2.89 (br d, J = 19.19 Hz, 1 H) 2.70 (s, 6 H) 2.46 - 2.34 (m, 1 H) 2.34 - 2.24 (m, 1 H) 1.97 - 1.83 (m, 2 H) 1.77 (br d, J= 5.50 Hz, 2 H). HPLC: 95.66% (220 nm), 95.74% (215 nm), 96.35% (254 nm). MS (ESI): mass calcd. C17H20CIN3O5S3 477.03 m/z found 477.8 [M+l] ⁺ .

Example 124: N5-[3-chloro-2-[2-(hydroxymethyl)-l-piperidyl] phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00584] Step 1. A mixture of 2-piperidylmethanol (722 mg, 6.27 mmol, 1 eq), 1 -chloro-2-fluoro- 3 -nitro-benzene (1.1 g, 6.27 mmol, 1 eq) , K ₂ CO ₃ (1.73 g, 12.5 mmol, 2 eq) and DIEA (1.62 g, 12.5 mmol, 2.18 mL, 2 eq) in dioxane (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 110 °C for 12 hours under N2 atmosphere. LC-MS showed 1 -chloro-2-fluoro- 3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. Then it was separated between water (20 mL) and EtOAc (40 mL). The organic phase was separated, washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-15% EtOAc /petroleum ether gradient @ 60 mL/min) to give desired [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] methanol (0.35 g, 1.29 mmol, 20.6% yield) as ayellow oil. MS (ESI): mass calcd. For C12H15CIN2O3 270.08 m/z found 271.1 [M+l] ⁺ .

[00585] Step 2: To a solution of [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] methanol (200 mg, 739 pmol, 1 eq) in EtOH (2 mL) and H ₂ O (0.5 mL) was added Fe (206 mg, 3.69 mmol, 5 eq) and AcOH (177 mg, 2.96 mmol, 169 μL, 4 eq). The mixture was stirred at 80 °C for 0.5 hour. LC-MS showed [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] methanol was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give desired [l-(2-amino-6-chloro-phenyl)-2-piperidyl] methanol (180 mg, crude) as ayellow oil. MS (ESI): mass calcd. For C12H17CIN2O 240.10 m/z found 241.2 [[M+l] ⁺ . [00586] Step 3. A mixture of [l-(2-amino-6-chloro-phenyl)-2-piperidyl] methanol (170 mg, 706 pmol, 1 eq) and 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (246 mg, 847 pmol, 1.2 eq) in Py (2 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 1 hour under N2 atmosphere. LCMS showed [l-(2-amino-6-chloro-phenyl)-2-piperidyl] methanol was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove Py. The residue was purified by prep-TLC (SiO2, petroleum ether/EtOAc = 1/1) to give desired N5-[3-chloro-2-[2- (hydroxymethyl)-l -piperidyl] phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (45.9 mg, 91.9 pmol, 13.0% yield, 98.9% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.27 (s, 1H), 7.76 (d, J= 4.0 Hz, 1H), 7.65 (d, J= 4.0 Hz, 1H), 7.51 (dd, J= 1.4, 8.0 Hz, 1H), 7.34 - 7.27 (m, 1H), 7.24 - 7.20 (m, 1H), 4.88 (br t, J= 4.4 Hz, 1H), 3.37 (br s, 1H), 3.20 - 3.08 (m, 2H), 2.92 - 2.84 (m, 1H), 2.63 (s, 6H), 1.77 (br s, 1H), 1.69 (br d, J = 11.1 Hz, 1H), 1.63 (br s, 2H), 1.46 (br d, J = 5.8 Hz, 1H), 1.35 - 1.26 (m, 2H). HPLC: 98.88% (220 nm), 97.81% (215 nm), 99.44% (254 nm). MS (ESI): mass calcd. For C18H24CIN3O5S3 493.06 m/z found 494.1 [M+l] ⁺ .

Example 125: N2, N2-dimethyl-N5-[2-(2-methyl-l-piperidyl) phenyl] thiophene-2, 5- disulfonamide

[00587] Step 1. A mixture of 1 -fluoro-2-nitro-benzene (2 g, 14.2 mmol, 1.49 mL, 1 eq) , 2- methylpiperidine (1.69 g, 17.0 mmol, 2.01 mL, 1.2 eq) , DIEA (3.66 g, 28.4 mmol, 4.94 mL, 2 eq) and K ₂ CO ₃ (3.92 g, 28.4 mmol, 2 eq) in dioxane (20 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 120 °C for 12 hours under N2 atmosphere. TLC indicated l-fluoro-2-nitro-benzene was consumed completely and two new spots formed. Then it was separated between 20 mL of H ₂ O and 40 mL of EtOAc. The organic phase was separated, washed with 30 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0~0% EtOAc/petroleum ether gradient @ 60 mL/min) to give desired 2-methyl-l -(2 -nitrophenyl)piperi dine (3 g, 13.6 mmol, 96.1% yield) as a yellow oil. [00588] Step 2: To a solution of 2-methyl-l -(2 -nitrophenyl)piperi dine (2.8 g, 12. mmol, 1 eq) in EtOH (30 mL) and H ₂ O (5 mL) was added Fe (3.55 g, 63.6 mmol, 5 eq) and AcOH (3.05 g, 50.9 mmol, 2.91 mL, 4 eq). The mixture was stirred at 80 °C for 0.5 hour. TLC indicated 2-methyl-l -(2- nitrophenyl) piperidine was consumed completely and one new spot formed. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a crude product. Then it was separated between 20 mL of NaOH (1 N) and 40 mL of ethyl acetate. The organic phase was separated, washed with 30 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 2-(2-methyl-l-piperidyl)aniline (2.3 g, crude) as a yellow oil.

[00589] Step 3. A mixture of compound 2-(2-methyl-l-piperidyl)aniline (1.14 g, 3.94 mmol, 1.5 eq) and 2-(2-methyl-l-piperidyl)aniline (500 mg, 2.63 mmol, 1 eq) in Py (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 1 hour under N2 atmosphere. LC-MS showed 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove Py. The residue was purified by prep-HPLC (neutral condition; according to LCMS); Method: column: Waters Xbridge C18 150*50mm* 10um;mobile phase: [water(10mM NH4HCOs)-ACN];B%: 50%-80%,10min) to give desired N2, N2-dimethyl-N5- [2-(2-methyl-l-piperidyl)phenyl]thiophene-2, 5 -disulfonamide (673 mg, 1.52 mmol, 57.8% yield, 100% purity) as a pale brown solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.25 (br s, 1H), 7.72 (d, J = 4.0 Hz, 1H), 7.63 (d, J= 4.0 Hz, 1H), 7.53 (d, J= 7.8 Hz, 1H), 7.30 (d, J= 7.5 Hz, 1H), 7.25 - 7.13 (m, 2H), 2.89 - 2.79 (m, 1H), 2.62 (s, 6H), 2.46 - 2.38 (m, 1H), 2.11 - 2.02 (m, 1H), 1.75 - 1.64 (m, 2H), 1.61 - 1.46 (m, 2H), 1.44 - 1.30 (m, 2H), 0.52 (d, J= 6.1 Hz, 3H). HPLC: 100.0% (220 nm), 100.0% (215 nm), 100.0% (254 nm). MS (ESI): mass calcd. For C18H25N3O4S3 443.1 m/z found 444.1 [M+H] ⁺ .

Example 126: N5-[2-(4, 4-dimethyl-l-piperidyl)-3, 5-difluoro-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00590] Step 1. A mixture of 2-bromo-l, 5 -difluoro-3 -nitro-benzene (600 mg, 2.52 mmol, 1 eq), 4, 4-dimethylpiperidine (528 mg, 3.53 mmol, 1.4 eq, HC1), CS ₂ CO ₃ (2.46 g, 7.56 mmol, 3 eq), Xantphos (72.9 mg, 126 pmol, 0.05 eq) and Pd(OAc)2 (28.3 mg, 126 pmol, 0.05 eq) in dioxane (5 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 80 °C for 12 hours under N2 atmosphere. LC-MS showed 2-bromo-l, 5 -difluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O 20 mL and extracted with EtOAc 30 mL (10 mL * 3). The combined organic layers were washed with brine 30 mL (10 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 5/1) to give desired l-(2, 4-difluoro-6-nitro-phenyl)-4, 4-dimethyl-piperidine (200 mg, 740 pmol, 29.4% yield) as a yellow oil. MS (ESI): mass calcd. For C13H16F2N2O2 270.12, m/z found 271.2 [M+H] ⁺ .

[00591] Step 2\ To a solution of l-(2, 4-difluoro-6-nitro-phenyl)-4, 4-dimethyl-piperidine (200 mg, 740 pmol, 1 eq) and in H ₂ O (1 mL) and EtOH (5 mL) was added Fe (207 mg, 3.70 mmol, 5 eq) and NH4CI (396 mg, 7.40 mmol, 10 eq). The mixture was stirred at 80 °C for 0.5 hour. LC-MS showed l-(2, 4-difluoro-6-nitro-phenyl)-4, 4-dimethyl-piperidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was fdtered and concentrated under reduced pressure to give a residue. The residue was diluted with H ₂ O 20 mL and extracted with EtOAc 90 mL (30 mL * 3). The combined organic layers were washed with brine 90 mL (30mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 5/1) to give desired 2-(4, 4- dimethyl-l-piperidyl)-3, 5 -difluoro-aniline (60 mg, 250 pmol, 33.7% yield) as a red oil. MS (ESI): mass calcd. For C13H18F2N2 240.14, m/z found 264.2 [M+H] ⁺ .

[00592] Step 3. To a solution of 2-(4, 4-dimethyl-l-piperidyl)-3, 5 -difluoro-aniline (60 mg, 250 pmol, 1 eq) in Py (1 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (101 mg, 350 pmol, 1.4 eq). The mixture was stirred at 15 °C for 1 hour. LC-MS showed 2-(4, 4-dimethyl-l- piperidyl)-3, 5 -difluoro-aniline was remained and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by /vcp-HPLC (TFA condition; column: Phenomen ex Luna Cl 8 75*30mm*3um; mobile phase: [water (0.2% FA)-ACN]; B%: 50%-80%,8 min) to give desired N5- [2-(4, 4-dimethyl-l-piperidyl)-3, 5 -difluoro-phenyl] -N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (32.2 mg, 63.8 pmol, 25.5% yield, 97.7% purity) as a pink solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.82 - 9.60 (m, 1H), 7.75 (d, J= 4.0 Hz, 1H), 7.67 (d, J= 4.1 Hz, 1H), 7.16 - 7.10 (m, 2H), 2.67 (s, 6H), 2.64 - 2.52 (m, 4H), 1.47 - 1.31 (m, 4H), 0.94 (s, 6H). HPLC: 97.74% (220 nm), 97.69% (215 nm), 98.70% (254 nm). MS (ESI): mass calcd. For C19H25F2N3O4S3 493.1 m/z found 494.1 [M+H] ⁺ .

Example 127: N5-[3-chloro-2-[2-(2-hydroxy ethyl)- 1-piperidyl] phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide [00593] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (2.99 g, 17.0 mmol, 1.1 eq) and 2- (2 -piperidyl) ethanol (2 g, 15.5 mmol, 1.98 mL, 1 eq) in dioxane (30 mL) was added K ₂ CO ₃ (4.28 g, 31.0 mmol, 2 eq) and DIEA (4.00 g, 31.0 mmol, 5.39 mL, 2 eq). The mixture was stirred at 120 °C for 12 hours. LCMS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed and desired mass was detected. The crude was added H ₂ O (40 mL), and extracted with EtOAc 90 mL (30 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 1/1 to 1/2) to give desired 2-[l-(2-chloro-6-nitro-phenyl)-2-piperidyl] ethanol (2 g, 7.02 mmol, 45.4% yield) as a brown oil. MS (ESI): mass calcd. For C13H17CIN2O3284.09 m/z found 285.2 [M+H] ⁺ .

[00594] Step 2: To a solution of 2-[l-(2-chloro-6-nitro-phenyl)-2-piperidyl] ethanol (500 mg, 1.76 mmol, 1 eq) in EtOH (5 mL) and H ₂ O (0.5 mL) was added Fe (490 mg, 8.78 mmol, 5 eq) and NH4CI (751 mg, 14.1 mmol, 8 eq). The mixture was stirred at 80 °C for 12 hours. LCMS showed 2- [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] ethanol was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude oil. The crude was added H ₂ O (40 mL), and extracted with EtOAc 90 mL (30 mL * 3). The combined organic layers were washed with brine 50 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 1/1 to 1/2) to give desired 2-[l-(2-amino-6-chloro- phenyl)-2-piperidyl] ethanol (300 mg, 1.18 mmol, 67.1% yield) as a brown oil. MS (ESI): mass calcd. For C13H19CIN2O 254.12 m/z found 255.2 [M+H] ⁺ .

[00595] Step 3: To a solution of 2-[l-(2-amino-6-chloro-phenyl)-2-piperidyl] ethanol (300 mg, 1.18 mmol, 1 eq) in Py (3 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (375 mg, 1.30 mmol, 1.1 eq). The mixture was stirred at 20 °C for 2 hours. LCMS showed 2-[l-(2- amino-6-chloro-phenyl)-2-piperidyl] ethanol was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini-NX 80*40mm*3um; mobile phase: [water (10mM NEEHCO3)-ACN]; B%: 32%-52%, 8min) to give desired N5-[3-chloro-2-[2-(2 -hydroxy ethyl)-l-piperidyl]phenyl]-N2,N2-dimethyl-thiophene-2, 5- disulfonamide (196 mg, 386 pmol, 32.8% yield, 99.9% purity) as a pale yellow solid. ¹ H NMR (400 MHz, DMSO-r/e) δ 7.54 - 7.48 (m, 2H), 7.20 (dd, J= 1.3, 8.2 Hz, 1H), 6.88 - 6.83 (m, 1H), 6.74 (dd, J= 1.3, 8.0 Hz, 1H), 4.15 - 4.07 (m, 1H), 3.90 - 3.83 (m, 1H), 3.51 - 3.43 (m, 1H), 3.43 - 3.36 (m, 1H), 3.21 - 3.12 (m, 1H), 2.66 (s, 6H), 2.58 - 2.54 (m, 1H), 2.14 - 2.04 (m, 1H), 1.99 - 1.87 (m, 2H), 1.85 - 1.74 (m, 2H), 1.64 - 1.45 (m, 3H). HPLC: 99.85% (220 nm), 99.83% (215 nm), 99.78% (254 nm). MS (ESI): mass calcd. For C19H26CIN3O5S3 507.07 m/z found 508.1 [M+H] ⁺ .

Example 128: N2, N2-dimethyl-N5-[2-[2-(2-morpholinoethyl)-l-piperidyl] phenyl] thiophene-2, 5-disulfonamide

[00596] Step 1. To a solution of l-fluoro-2-nitro-benzene (100 mg, 709 pmol, 74.6 μL, 1 eq) and 2-(2-piperidyl)ethanol (128 mg, 992 pmol, 127 μL, 1.4 eq) in dioxane (2 mL) was added K ₂ CO ₃ (196 mg, 1.42 mmol, 2 eq) and DIEA (183 mg, 1.42 mmol, 247 μL, 2 eq). The mixture was stirred at 120 °C for 12 hours. LC-MS showed 1 -fluoro-2-nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O 30 mL and extracted with EtOAc 150 mL (50 mL * 3). The combined organic layers were washed with brine 30 mL, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 2/1) to give desired 2-[l-(2-nitrophenyl)-2-piperidyl] ethanol (100 mg, 400 pmol, 56.4% yield) as a red oil. MS (ESI): mass calcd. For C13H18N2O3 250.13 m/z found 251.2 [M+H] ⁺ .

[00597] Step 2\ To a solution of 2-[l-(2-nitrophenyl)-2-piperidyl]ethanol (2.1 g, 8.39 mmol, 1 eq) in DCM (20 mL) was added TEA (1.70 g, 16.8 mmol, 2.34 mL, 2 eq) and MsCl (1.44 g, 12.6 mmol, 974 pl, 1.5 eq) at 0 °C. The mixture was stirred at 20 °C for 1 hour. LC-MS showed 2-[l-(2- nitrophenyl)-2-piperidyl] ethanol was consumed completely and one main peak with desired mass was detected. The reaction mixture was quenched by addition Saturated sodium bicarbonate aqueous solution at 0 °C and extracted with DCM 90 mL (30 mL * 3). The combined organic layers were washed with brine 50 mL dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 2-[l-(2-nitrophenyl)-2-piperidyl]ethyl methanesulfonate (3 g, crude) as a yellow oil. MS (ESI): mass calcd. For C14H20N2O5S 328.11 m/z found 329.2 [M+H] ⁺ . [00598] Step 3. To a solution of 2-[l-(2-nitrophenyl)-2-piperidyl]ethyl methanesulfonate (1 g, 3.05 mmol, 1 eq) in THF (10 mL) was added TEA (924 mg, 9.14 mmol, 1.27 mL, 3 eq) and morpholine (1.33 g, 15.2 mmol, 1.34 mL, 5 eq). The mixture was stirred at 70 °C for 12 hours. LC- MS showed 2-[l-(2-nitrophenyl)-2-piperidyl] ethyl methanesulfonate was consumed completely and one main peak with desired mass was detected. The residue was diluted with H ₂ O 100 mL and extracted with EtOAc 300 mL (100 mL * 3). The combined organic layers were washed with brine 300 mL (lOOmL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 1/0 to 2/1) to give desired 4-[2-[l-(2-nitrophenyl)-2-piperidyl] ethyl] morpholine (950 mg, 2.97 mmol, 97.7% yield) as a yellow oil. MS (ESI): mass calcd. For C17H25N3O3319.19 m/z found 320.3 [M+H] ⁺ .

[00599] Step 4: To a solution of 4-[2-[l-(2-nitrophenyl)-2-piperidyl] ethyl] morpholine (100 mg, 313 pmol, 1 eq) in H ₂ O (0.4 mL) and EtOH (2 mL) as added Fe (87.4 mg, 1.57 mmol, 5 eq) and NH4CI (167 mg, 3.13 mmol, 10 eq). The mixture was stirred at 80 °C for 1 hour. LC-MS showed 4- [2-[l-(2-nitrophenyl)-2-piperidyl] ethyl] morpholine was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was diluted with H ₂ O 10 mL and extracted with EtOAc 60 mL (20mL * 3). The combined organic layers were washed with brine 60 mL (20mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether: EtOAc = 3:1) to give desired 2-[2-(2- morpholinoethyl)-! -piperidyl] aniline (75 mg, 259.14 pmol, 82.8% yield) as a brown oil. MS (ESI): mass calcd. For C17H27N3O 502.08 m/z found 503.1 [M+H] ⁺ .

[00600] Step 5: To a solution of 2-[2-(2-morpholinoethyl)-l-piperidyl] aniline (250 mg, 864 pmol, 1 eq) in Py (3 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (375 mg, 1.30 mmol, 1.5 eq). The mixture was stirred at 15 °C for 1 hour. LC-MS showed 2-[2-(2- morpholinoethyl)-! -piperidyl] aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition; column: Phenomenex Gemini-NX C18 75*30mm*3um; mobile phase: [water (0.05%NH3H20+10mM NEEHCC^-ACN]; B%: 30%-70%, 8min) to give desired N2, N2-dimethyl-N5-[2-[2-(2-morpholinoethyl)-l- piperidyl]phenyl]thiophene-2, 5 -disulfonamide (192 mg, 354 pmol, 41.0% yield, 100% purity) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.76 - 7.72 (m, 1H), 7.66 - 7.62 (m, 1H), 7.53 - 7.49 (m, 1H), 7.29 - 7.29 (m, 1H), 7.31 - 7.26 (m, 1H), 7.22 - 7.12 (m, 2H), 3.48 (t, J= 4.5 Hz, 4H), 2.91 (br s, 1H), 2.66 (s, 6H), 2.50 - 2.44 (m, 1H), 2.20 (br d, J= 11.7 Hz, 1H), 2.15 - 2.06 (m, 5H), 2.04 - 1.95 (m, 1H), 1.88 - 1.79 (m, 1H), 1.77 - 1.67 (m, 1H), 1.63 - 1.47 (m, 2H), 1.44 - 1.33 (m, 2H), 1.15 - 1.06 (m, 2H). HPLC: 100.0% (220 nm), 100.0% (215 nm), 100.0% (254 nm). MS (ESI): mass calcd. For C23H34N4O5S3542.17 m/z found 543.2 [M+H] ⁺ .

Example 129: N5-[3-chloro-2-[2-(cy anomethyl)- 1-piperidyl] phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00601] Step 1. To a solution of 1 -bromo-2-fluoro-3 -nitro-benzene (2 g, 9.09 mmol, 1 eq) and 2- piperidylmethanol (1.15 g, 10.0 mmol, 1.1 eq) in 1,4-dioxane (10 mL) was added K ₂ CO ₃ (2.51 g, 18.2 mmol, 2 eq) and DIEA (2.35 g, 18.2 mmol, 3.17 mL, 2 eq). The mixture was stirred at 100 °C for 12 hours. LC-MS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O 20mL and extracted with EtOAc 150 mL (50mL * 3). The combined organic layers were washed with brine 60 mL (20mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0-20% EtOAc /petroleum ether gradient @ 80 mL/min) to give desired [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] methanol (1 g, 3.17 mmol, 34.9% yield) as a yellow oil. MS (ESI): mass calcd. For C12H15CIN2O3 270.08 m/z found 271.1 [M+H] ⁺ .

[00602] Step 2: To a solution of [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] methanol (600 mg, 2.22 mmol, 1 eq) in DCM (10 mL) was added TEA (449 mg, 4.43 mmol, 617 μL, 2 eq) at 0 °C. Then MsCl (381 mg, 3.32 mmol, 257 μL, 1.5 eq) was added dropwise o the mixture at 0 °C over 10 mins. After addition, the mixture was stirred at 20 °C for 1 hour. LC-MS showed [l-(2-chloro-6- nitro-phenyl)-2-piperidyl] methanol was consumed completely and one main peak with desired mass was detected. The reaction mixture was quenched by addition Saturated sodium bicarbonate aqueous solution at 0 °C and extracted with DCM 60 mL (20 mL * 3). The combined organic layers were washed with brine 60 mL (20 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give desired [l-(2-chloro-6-nitro-phenyl)-2-piperidyl]methyl methanesulfonate (700 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C13H17CIN2O5S 348.05 m/z found 349.0 [M+H] ⁺ .

[00603] Step 3: To a solution of [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] methyl methanesuLfonate (300 mg, 860 pmol, 1 eq) in DMF (3 mL) was added TEA (261 mg, 2.58 mmol, 359.14 μL, 3 eq) and NaCN (63.2 mg, 1.29 mmol, 1.5 eq). The mixture was stirred at 80 °C for 1 hour. LC-MS showed [l-(2-chloro-6-nitro-phenyl)-2-piperidyl] methyl methanesuLfonate was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (5 mL) and extracted with EtOAc (5 mL * 2). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20g SepaFlash® Silica Flash Column, Eluent of 0~20 % EtOAc /petroleum ether gradient @ 40 mL/min) to give desired 2-[l-(2-chloro-6-nitro-phenyl)-2-piperidyl] acetonitrile (100 mg, 357 pmol, 41.6% yield) as ayellow oil. MS (ESI): mass calcd. For C13H14CIN3O2 279.08 m/z found 280.0 [M+H] ⁺ .

[00604] Step 4. To a solution of 2-[l-(2-chloro-6-nitro-phenyl)-2-piperidyl] acetonitrile (100 mg, 358 pmol, 1 eq) in EtOH (3 mL) and H ₂ O (0.6 mL) was added Fe (200 mg, 3.57 mmol, 10 eq), AcOH (85.9 mg, 1.43 mmol, 81.78μL, 4 eq) and NH4CI (191 mg, 3.57 mmol, 10 eq). The mixture was stirred at 80 °C for 2 hours. LC-MS showed 2-[l-(2-chloro-6-nitro-phenyl)-2-piperidyl] acetonitrile was consumed completely and one main peak with desired mass was detected. The reaction mixture was adjusted to pH = 9 with Saturated sodium hydroxide aqueous solution at 20 °C, and then diluted with H ₂ O 10 mL and extracted with EtOAc 30 mL (10 mL * 3). The combined organic layers were washed with brine 10 mL (5 mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0-20% EtOAc /petroleum ether gradient @ 30 mL/min) to give desired 2-[l-(2-amino-6-chloro-phenyl)-2- piperidyl] acetonitrile (60 mg, 240 pmol, 67.2% yield) as ayellow oil. mass calcd. For C13H16CIN3 249.10 m/z found 250.0 [M+H] ⁺ . [00605] Step 5: To a solution of 2-[l-(2-amino-6-chloro-phenyl)-2-piperidyl] acetonitrile (15 mg, 60.1 pmol, 1 eq) and 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (22.6 mg, 78.1 pmol, 1.3 eq) in pyridine (0.5 mL). The mixture was stirred at -10 °C for 1 hour. LC-MS showed 2-[l-(2- amino-6-chloro-phenyl)-2-piperidyl] acetonitrile was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H ₂ O (3 mL) and extracted with EtOAc (2 mL * 3). The combined organic layers were washed with brine 4 (2 mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (FA condition: column: Phenomenex Luna Cl 8 150*30mm*5um;mobile phase: [water(0.2%FA)-ACN];B%: 55%-85%,8min) to give desired N5- [3 -chloro-2-[2-(cy anomethyl)- 1- piperidyl]phenyl]-N2, N2-dimethyl-thiophene-2, 5-disulfonamide (2 mg, 3.92 pmol, 6.52% yield, 98.5% purity) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.22 (s, 1H), 7.78 (d, J= 4.0 Hz, 1H), 7.67 (d, J= 4.0 Hz, 1H), 7.53 (dd, J= 1.3, 8.2 Hz, 1H), 7.34 (t, J= 8.2 Hz, 1H), 7.24 (dd, J = 1.4, 8.1 Hz, 1H), 3.79 - 3.70 (m, 1H), 3.10 - 3.00 (m, 1H), 2.72 (s, 1H), 2.66 - 2.62 (m, 6H), 2.34 - 2.28 (m, 2H), 1.81 - 1.77 (m, 2H), 1.64 (br d, J= 10.8 Hz, 1H), 1.47 (br s, 2H), 1.41 - 1.29 (m, 1H). HPLC: 98.58% (220 nm), 96.85% (215 nm), 99.53% (254 nm). MS (ESI): mass calcd. For C19H23CIN4O4S3 502.06 m/z found 503.1 [M+H] ⁺ .

Example 130: N2, N2-dimethyl-N5-[2-[2-(morpholinomethyl)-l-piperidyl] phenyl] thiophene-2, 5-disulfonamide

[00606] Step 1. To a solution of l-fluoro-2-nitro-benzene (4 g, 28.4 mmol, 2.99 mL, 1 eq) and 2- piperidylmethanol (3.92 g, 34.0 mmol, 1.2 eq) in dioxane (40 mL) was added K ₂ CO ₃ (7.84 g, 56.7 mmol, 2 eq) and DIEA (7.33 g, 56.7 mmol, 9.88 mL, 2 eq). The mixture was stirred at 120 °C for 12 hours. LCMS showed 1 -fluoro-2-nitro-benzene was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude oil. The crude was added H ₂ O (50 mL), and extracted with EtOAc 150 mL (50 mL * 3). The combined organic layers were dried over ISfeSCE, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 10/1 to 5/1) to give desired [l-(2-nitrophenyl)-2-piperidyl] methanol (3.65 g, 15.5 mmol, 54.5% yield) as a red oil. MS (ESI): mass calcd. For C12H16N2O3 236.12 m/z found 237.2 [M+H] ⁺ .

[00607] Step 2\ To a solution of [l-(2-nitrophenyl)-2-piperidyl] methanol (3.65 g, 15.5 mmol, 1 eq) in DCM (40 mL) was added dropwise TEA (3.13 g, 30.9 mmol, 4.30 mL, 2 eq) at 0 °C. After addition, the mixture was stirred at this temperature for 0.5 hour, and then MsCl (2.65 g, 23.2 mmol, 1.79 mL, 1.5 eq) was added drop wise at 0 °C. The resulting mixture was stirred at 20 °C for 11.5 hours. LCMS showed [l-(2-nitrophenyl)-2-piperidyl] methanol was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired [l-(2-nitrophenyl)-2-piperidyl] methyl methanesulfonate (5 g, crude) as a brown oil. MS (ESI): mass calcd. For C13H18N2O5S 314.09 m/z found 315.2 [M+H] ⁺ . [00608] Step 3: To a solution of [l-(2-nitrophenyl)-2-piperidyl]methyl methanesulfonate (1 g,

3.18 mmol, 1 eq) in THF (10 mL) was added TEA (966 mg, 9.54 mmol, 1.33 mL, 3 eq) and morpholine (1.39 g, 15.9 mmol, 1.40 mL, 5 eq). The mixture was stirred at 70 °C for 12 hours. LCMS showed [l-(2-nitrophenyl)-2-piperidyl] methyl methanesulfonate was consumed completely and desired mass was detected. The crude was added H ₂ O (10 mL), and extracted with EtOAc 30 mL (10 mL * 3). The combined organic layers were washed with brine 5 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/EtOAc = 5/1 to 4/1) to give desired 4-[[l-(2-nitrophenyl)-2- piperidyl] methyl] morpholine (360 mg, 1.18 mmol, 37.1% yield) as a brown oil. MS (ESI): mass calcd. For C16H23N3O3 305.17 m/z found 306.3 [M+H] ⁺ .

[00609] Step 4. To a solution of 4-[[l-(2-nitrophenyl)-2-piperidyl] methyl] morpholine (360 mg,

1.18 mmol, 1 eq) in EtOH (4 mL) and H ₂ O (1 mL) was added Fe (329 mg, 5.89 mmol, 5 eq) and NH4CI (504 mg, 9.43 mmol, 8 eq). The mixture was stirred at 80 °C for 2 hours. LC-MS showed 4- [[l-(2-nitrophenyl)-2-piperidyl] methyl] morpholine was consumed and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give desired 2- [2-(morpholinomethyl)-l -piperidyl] aniline (350 mg, crude) as a brown oil. MS (ESI): mass calcd. For C16H25N3O 275.2 m/z found 276.3 [M+H] ⁺ . [00610] Step 5: To a solution of 2- [2-(morpholino methyl)-! -piperidyl] aniline (280 mg, 1.02 mmol, 1 eq) in Py (2 mL) was added 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (294 mg, 1.02 mmol, 1 eq). The mixture was stirred at 15 °C for 2 hours. LC-MS showed 2-[2- (morpholinomethyl)-l -piperidyl] aniline was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (neutral condition, LCMS, column: Phenomenex Gemini-NX C18 75*30mm*3um; mobile phase: [water (10mM NH4HCOs)-ACN]; B%: 25%-55%, lOmin) to give desired N2, N2-dimethyl-N5- [2- [2-(morpholinomethyl)-l -piperidyl] phenyl] thiophene-2, 5-disulfonamide (85.1 mg, 155 pmol, 15.3% yield, 96.5% purity) as a yellow solid. ^J H NMR (400 MHz, DMSO-d ₆ ,) δ 7.63 (q, J= 3.8 Hz, 2H), 7.43 (br d, J= 7.5 Hz, 1H), 7.30 (br d, J = 6.4 Hz, 1H), 7.12 - 7.06 (m, 2H), 3.55 (br d, J= 3.7 Hz, 4H), 2.70 - 2.68 (m, 1H), 2.66 (s, 6H), 2.60 - 2.57 (m, 2H), 2.54 (br s, 2H), 2.43 (br s, 1H), 2.44 - 2.40 (m, 1H), 2.36 - 2.34 (m, 1H), 2.36 - 2.34 (m, 1H), 2.31 - 2.26 (m, 2H), 2.18 - 2.12 (m, 1H), 2.10 (s, 1H), 1.85 - 1.78 (m, 1H), 1.72 - 1.64 (m, 1H), 1.60 - 1.51 (m, 2H), 1.46 - 1.36 (m, 2H). HPLC: 99.97% (220 nm), 99.97% (215 nm), 99.97% (254 nm).MS (ESI): mass calcd. For C22H32N4O5S3 528.15 m/z found 529.2 [M+H] ⁺ .

Example 131: N5-[3-chloro-2-[2-(methoxymethyl)-4, 4-dimethyl-l-piperidyl] phenyl] -N2, N2- dimethyl-thiophene-2, 5-disulfonamide

[00611] Step 1. To a solution of 4, 4-dimethylpiperidine (5 g, 33.4 mmol, 1 eq, HC1) in DCM (50 mL) was added DMAP (408.15 mg, 3.34 mmol, 0.1 eq) and TEA (6.76 g, 66.8 mmol, 9.30 mL, 2 eq) at 0 °C. The mixture was stirred at 0 °C for 10 mins. Then BOC2O (8.02 g, 36.8 mmol, 8.44 mL, 1.1 eq) was added to the above mixture. The mixture was stirred at 20 °C for 1 hour. LC-MS showed 4, 4-dimethylpiperidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O 30 mL and extracted with DCM 150 mL (50 mL * 3). The combined organic layers were washed with brine 60 mL (30 mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0-15% EtOAc /petroleum ether gradient @ 80 mL/min) to give desired tert-butyl 4, 4- dimethylpiperidine-1 -carboxylate (7 g, 32.8 mmol, 98.2% yield) as a colorless oil. MS (ESI): mass calcd. For C12H23NO2 213.17 m/z found 158.1 [M-56H] ⁺ .

[00612] Step 2: A mixture of tert-butyl 4,4-dimethylpiperidine-l -carboxylate (7 g, 32.8 mmol, 1 eq) in THF (70 mL) was cooled to -78 °C and treated with TMEDA (3.81 g, 32.8 mmol, 4.95 mL, 1 eq) followed by slow addition of H2CO3 (14.0 g, 226 mmol, 6.88 eq) over a 30 minute period. The mixture was stirred at -78 °C for 0.5 hour. TLC (petroleum ether/EtOAc = 1/1) indicated tert-butyl 4, 4-dimethylpiperidine- 1 -carboxylate was consumed completely and one new spot formed. The reaction was clean according to TLC. The reaction mixture was diluted with H ₂ O 50 mL and extracted with EtOAc 300 mL (100 mL * 3). The combined organic layers were washed with brine 150 mL (50 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give desired l-tert-butoxycarbonyl-4, 4-dimethyl-piperidine-2-carboxylic acid (4.88 g, 19.0 mmol, 58.0% yield) as a yellow oil.

[00613] Step 3. A mixture of 1 -tert-butoxy carbonyl-4, 4-dimethyl-piperidine-2-carboxylic acid (4.88 g, 19.0 mmol, 1 eq) in THF (50 mL) was degassed and purged with N2 for 3 times, and then borane /tetrahydrofuran (1 M, 56.9 mL, 3 eq) was added dropwise at 0 °C and the mixture was stirred at 20 °C for 12 hours under N2 atmosphere. LC-MS showed l-tert-butoxycarbonyl-4, 4-dimethyl- piperidine-2-carboxylic acid was consumed completely and one main peak with desired mass was detected. The reaction mixture was quenched by addition H ₂ O 20 mL at 0°C, and then diluted with brine 20 mL and extracted with EtOAc 150 mL (50 mL * 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0-20% EtOAc /petroleum ether gradient @ 80 mL/min) to give desired tert-butyl 2-(hydroxymethyl)-4, 4-dimethyl-piperidine-l -carboxylate (1.5 g, 6.16 mmol, 32.5% yield) as a colorless oil. MS (ESI): mass calcd. For C13H25NO3 243.18 m/z found 188.1 [M-56H] ⁺ .

[00614] Step 4. To a solution of tert-butyl 2-(hydroxymethyl)-4,4-dimethyl-piperidine-l- carboxylate (1.5 g, 6.16 mmol, 1 eq) in THF (30 mL) was added NaH (986 mg, 24.7 mmol, 60% purity, 4 eq) at 0 °C, the mixture was stirred at 0 °C for 0.5 hour. Then Mel (3.50 g, 24.7 mmol, 1.53 mL, 4 eq) was added drop wise to the above mixture at 0 °C over 10 mins. The mixture was stirred at 20 °C for 12 hours. LC-MS showed tert-butyl 2-(hydroxymethyl)-4, 4-dimethyl-piperidine- 1- carboxylate was consumed completely and one main peak with desired mass was detected. The reaction mixture was quenched by addition H ₂ O 20 mL at 0 °C, and extracted with EtOAc 90 mL (30 mL * 3). The combined organic layers were washed with brine 30 mL (10 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-10% EtOAc /petroleum ether gradient @ 60 mL/min) to give desired tert-butyl 2-(methoxymethyl)-4, 4-dimethyl-piperidine-l -carboxylate (440 mg, 1.71 mmol, 27.7% yield) as a yellow oil. MS (ESI): mass calcd. For C14H27NO3 257.2 m/z found 202.1 [M- 56H] ⁺ .

[00615] Step 5: To a solution of tert-butyl 2-(methoxymethyl)-4, 4-dimethyl-piperidine- 1- carboxylate (440 mg, 1.71 mmol, 1 eq) was added HCl/MeOH (4 M, 10.2 mL, 23.8 eq). The mixture was stirred at 20 °C for 0.5 hour. TLC (petroleum ether/EtOAc = 4/1) indicated tert-butyl 2- (methoxymethyl)-4, 4-dimethyl-piperidine-l -carboxylate was consumed completely and one new spot formed. The reaction was clean according to TLC. The reaction mixture was concentrated under reduced pressure to give desired 2-(methoxymethyl)-4, 4-dimethyl-piperidine (330 mg, crude, HC1) as a white solid was used directly for the next step without further purification.

[00616] Step 6: A mixture of 2-bromo-l-chloro-3 -nitro-benzene (120 mg, 508 pmol, 1 eq), 2- (methoxymethyl)-4, 4-dimethyl-piperidine (118 mg, 609 pmol, 1.2 eq, HC1), CS ₂ CO ₃ (496 mg, 1.52 mmol, 3 eq), CuiBuCx2 (20.2 mg, 50.8 pmol, 0.1 eq) in DMSO (1 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 120 °C for 12 hours under N2 atmosphere. LC-MS showed -60% of 2-(methoxymethyl)-4, 4-dimethyl-piperidine remained. Several new peaks were shown on LC-MS and -10% of desired compound was detected. The reaction mixture was diluted with H ₂ O 5 mL and extracted with EtOAc 15 mL (5 mL * 3). The combined organic layers were washed with brine 6 mL (3 mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 25 g SepaFlash® Silica Flash Column, Eluent of 0-10% EtOAc /petroleum ether gradient @ 35 mL/min) to give desired 1 -(2-chloro-6-nitro-phenyl)-2- (methoxymethyl)-4, 4-dimethyl-piperidine (38 mg, 121 pmol, 12.0% yield) as a yellow oil. MS (ESI): mass calcd. For C15H21CIN2O3 312.12 m/z found 313.1 [M+H] ⁺ .

[00617] Step 7: To a solution of l-(2-chloro-6-nitro-phenyl)-2-(methoxymethyl)-4, 4-dimethyl- piperidine (38 mg, 121 pmol, 1 eq) in EtOH (0.6 mL) and H ₂ O (0.15 mL) was added Fe (67.8 mg, 1.21 mmol, 10 eq) and NH4CI (65.0 mg, 1.21 mmol, 10 eq). The mixture was stirred at 80 °C for 1 hour. LC-MS showed l-(2-chloro-6-nitro-phenyl)-2-(methoxymethyl)-4, 4-dimethyl-piperidine was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O 2 mL and extracted with EtOAc (3 mL * 3). The combined organic layers were washed with brine (3 mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0-15% Ethyl acetate/petroleum ether gradient @ 20 mL/min) to give desired 3-chloro-2-[2-(methoxymethyl)-4, 4-dimethyl-l -piperidyl] aniline (26 mg, 91.9 pmol, 75.7% yield) as a yellow oil. MS (ESI): mass calcd. For C15H21CIN2O3 312.12 m/z found 313.1 [M+H] ⁺ .

[00618] Step 8: To a solution of 3-chloro-2-[2-(methoxymethyl)-4, 4-dimethyl-l -piperidyl] aniline (26 mg, 92.0 pmol, 1 eq) in pyridine (0.3 mL) was added 5 -(dimethylsulfamoyl) thiophene-2- sulfonyl chloride (40.0 mg, 138 pmol, 1.5 eq). The mixture was stirred at 20 °C for 1 hour. LC-MS showed 3-chloro-2-[2-(methoxymethyl)-4, 4-dimethyl-l -piperidyl] aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H ₂ O 1 mL and extracted with EtOAc 3 mL (1 mL * 3). The combined organic layers were washed with brine 2 mL (1 mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (FA condition: column: Phenomen ex Luna Cl 8 75*30mm*3um;mobile phase: [water(0.2%FA)-ACN];B%: 90%-98%,8min) to give desired N5-[3- chloro-2-[2-(methoxymethyl)-4,4-dimethyl-l-piperidyl]phenyl] -N2, N2-dimethyl-thiophene-2, 5- disulfonamide (13.6 mg, 25.3 pmol, 27.5% yield, 99.6% purity) as a yellow solid ¹ H NMR (400MHz, DMSO-r/e) δ 9.01 (s, 1H), 7.79 (d, J= 4.0 Hz, 1H), 7.67 (d, J= 4.0 Hz, 1H), 7.56 (dd, J = 1.4, 8.1 Hz, 1H), 7.39 - 7.31 (m, 1H), 7.29 - 7.23 (m, 1H), 3.68 (br dd, J= 2.9, 11.7 Hz, 1H), 3.37 (br d, J= 3.2 Hz, 1H), 2.98 - 2.94 (m, 1H), 2.93 (s, 3H), 2.73 (dd, J= 4.0, 10.0 Hz, 1H), 2.62 (s, 6H), 1.59 - 1.51 (m, 1H), 1.42 - 1.28 (m, 3H), 1.14 (br d, J= 10.8 Hz, 1H), 0.96 (d, J= 3.7 Hz, 6H). HPLC: 99.58% (220 nm), 99.79% (215 nm), 100.0% (254 nm). MS (ESI): mass calcd. For C21H30CIN3O5S3 535.1 m/z found 536.0 [M+H] ⁺ .

Example 132: N-[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] -5-(3-methoxyoxetan-3-yl) thiophene-2-sulfonamide

[00619] Step 1. To a solution of 2, 5 -dibromo thiophene (3 g, 12.4 mmol, 1.40 mL, 1 eq) in THF (30 mL) was added w-BuLi (2.5 M, 4.96 mL, 1 eq). The mixture was stirred at -78 °C for 30 mins. oxetan-3-one (983 mg, 13.6 mmol, 1.1 eq) was added to the mixture under N2. The mixture was stirred at -78 °C for 1 hour. TLC (petroleum ether/EtOAc = 3/1) indicated 2, 5 -dibromothiophene was consumed completely and one new spot formed. The reaction was clean according to TLC. The reaction mixture was quenched by addition sat. NH4CI aq. 40 mL, and extracted with EtOAc 40mL (10 mL *4). The combined organic layers were washed with brine 20 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 3-(5-bromo-2-thienyl)oxetan-3-o (3.7 g, crude) as a pink solid.

[00620] Step 2\ To a solution of 3-(5-bromo-2-thienyl) oxetan-3-ol (3.7 g, 15.74 mmol, 1 eq) in THF (50 mL) was added NaH (1.89 g, 47.21 mmol, 60% purity, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 0.5 hour. Mel (6.70 g, 47.2 mmol, 2.94 mL, 3 eq) was added to the mixture at 0 °C. The mixture was stirred at 15 °C for 12 hours. TLC (petroleum ether/EtOAc = 3/1) indicated 3- (5-bromo-2-thienyl) oxetan-3-ol 2 was consumed completely and one new spot formed. The reaction was clean according to TLC. The crude was added H ₂ O (200 mL), and extracted with EtOAc 300 mL (100 mL * 3). The combined organic layers were washed with brine 100 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 3-(5-bromo-2-thienyl)-3-methoxy- oxetane (2.8 g, crude) as an orange oil. [00621] Step 3: A mixture of phenylmethanethiol (548 mg, 4.42 mmol, 517 μL, 1.1 eq), 3-(5- bromo-2-thienyl)-3-methoxy-oxetane (1 g, 4.01 mmol, 1 eq), DIEA (1.04 g, 8.03 mmol, 1.40 mL, 2 eq), Xantphos (232 mg, 401 pmol, 0.1 eq) and Pd(dppf)C12 (73.4 mg, 100 pmol, 0.025 eq) in Tol. (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 110 °C for 12 hours under N2 atmosphere. TLC (petroleum ether/EtOAc = 3/1) indicated 3-(5-bromo-2-thienyl)-3- methoxy-oxetane was consumed completely and one new spot formed. The reaction mixture was concentrated under reduced pressure to remove Tol. The mixture was cooled at 20 °C and added H ₂ O (100 mL), and extracted with EtOAc 300 mL (100 mL * 3). The combined organic layers were washed with brine 50 mL, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/EtOAc = 10/1 to 1/1) to give desired 3-(5-benzylsulfanyl-2-thienyl)-3-methoxy-oxetane (880 mg, crude) was obtained as a yellow oil.

[00622] Step 4. To a solution of 3-(5-benzylsulfanyl-2-thienyl)-3-methoxy-oxetane (440 mg, 1.50 mmol, 1 eq) in AcOH (4 mL) and H ₂ O (1 mL) was added NCS (602.77 mg, 4.51 mmol, 3 eq). The mixture was stirred at 20 °C for 12 hours. TLC (petroleum ether/EtOAc = 3/1) showed 3-(5- benzylsulfanyl-2-thienyl)-3-methoxy-oxetane was consumed completely and one major new spot with larger polarity was detected. The reaction mixture was diluted with water 30 mL and extracted with EtOAc 90 mL (30 mL * 3). The combined organic layers were washed with brine 20 mL, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 5-(3- methoxyoxetan-3-yl)thiophene-2-sulfonyl chloride (400 mg, crude) as a yellow oil.

[00623] Step 5: To a solution of 5-(3-methoxyoxetan-3-yl)thiophene-2-sulfonyl chloride (169 mg, 628 pmol, 1.5 eq) in Py (1 mL) was added 3-chloro-2-(4,4-dimethyl-l-piperidyl)aniline (100 mg, 419 pmol, 1 eq). The mixture was stirred at 15 °C for 2 hours. LC-MS showed 5-(3-methoxyoxetan- 3-yl) thiophene-2-sulfonyl chloride was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (basic condition, column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water(0.05%NH ₃ H ₂ 0+10mM NH4HCO ₃ )-ACN];B%: 50%- 80%,8min) to give desired N-[3-chloro-2-(4,4-dimethyl-l-piperidyl)phenyl]-5-(3-methoxy oxetan-3- yl)thiophene-2-sulfonamide (46.1 mg, 96.7 pmol, 23.1% yield, 98.8% purity) as ayellow solid. ^J H NMR (400 MHz, DMSO-d6 ) δ 9.12 (s, 1 H), 7.54 (d, J= 3.91 Hz, 1 H), 7.43- 7.36 (m, 2 H), 7.26 - 7.21 (m, 2 H), 4.79 (d, J= 7.46 Hz, 2 H), 4.65 (d, J= 7.34 Hz, 2 H), 3.36 - 3.26 (m, 2 H), 3.12 (s, 3 H), 2.25 (br s, 2 H), 1.51 (br s, 2 H), 1.40 - 1.23 (m, 2 H), 0.98 (br s, 6 H). HPLC: 98.78% (220 nm), 98.83% (215 nm), 99.76% (254 nm). MS (ESI): mass calcd. For C21H27CIN2O4S2 470.11 m/z found 471.1 [M+H] ⁺ .

Example 133: N5-[3-chloro-2-[2-(2-methoxy ethyl)- 1-piperidyl] phenyl] -N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00624] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (45 mg, 256 pmol, 1 eq) in dioxane (0.5 mL) was added K ₂ CO ₃ (70.9 mg, 513 pmol, 2 eq) DIEA (66.3 mg, 513 pmol, 89.3 μL, 2 eq) and 2-(2-methoxyethyl) piperidine (50.7 mg, 282 pmol, 1.1 eq, HC1). The mixture was stirred at 120 °C for 12 hours. LC-MS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed completely and desired mass was detected. The reaction mixture was added to H ₂ O (30 mL), the aqueous phase was extracted with EtOAc (20 mL*3). The organic layer was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiC>2, petroleum ether/Ethyl acetate=l/O to 1/1) to give desired l-(2-chloro-6-nitro-phenyl)-2-(2- methoxyethyl) piperidine (110 mg, 368 pmol, 14.4% yield) as a yellow oil. MS (ESI): mass calcd. For C14H19CIN2O3 298.11 m/z found 299.2 [[M+H] ⁺ .

[00625] Step 2'. To a solution of l-(2-chloro-6-nitro-phenyl)-2-(2-methoxyethyl) piperidine (110 mg, 368 pmol, 1 eq) in EtOH (2 mL) and H ₂ O (0.2 mL) was added Fe (103 mg, 1.84 mmol, 5 eq) and NH4CI (197 mg, 3.68 mmol, 10 eq). The mixture was stirred at 80 °C for 0.5 hour. LC-MS showed l-(2-chloro-6-nitro-phenyl)-2-(2-methoxy ethyl) piperidine was consumed completely and desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The reaction mixture was added to H ₂ O (20 mL), the aqueous phase was extracted with EtOAc (10 mL*3). The organic layer was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated to give desired 3 -chloro-2-[2-(2-methoxy ethyl)- 1 -piperidyl] aniline (90 mg, crude) as an orange oil. MS (ESI): mass calcd. For C14H21CIN2O 268.13 m/z found 269.3 [M+H] ⁺ .

[00626] Step 3. To a solution of 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (129 mg, 446 pmol, 1.5 eq) in Py (1 mL) was added 3 -chloro-2-[2-(2-methoxy ethyl)- 1 -piperidyl] aniline (80 mg, 298 pmol, 1 eq). The mixture was stirred at 15 °C for 2 hours. LC-MS showed 3-chloro-2-[2- (2-methoxy ethyl)- 1 -piperidyl] aniline was consumed completely and desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-WLC (neutral condition, column: Phenomenex Gemini - NX C18 75*30mm*3um; mobile phase: [water (10mM NH4HCOs)-ACN]; B%: 45%-65%,8min) to give desired N5-[3-chloro-2-[2-(2-methoxyethyl)-l-piperidyl]phenyl]-N2,N2 -dimethyl-thiophene- 2, 5 -disulfonamide (42.3 mg, 78.9 pmol, 26.5% yield, 97.4% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.31 (s, 1 H), 7.69 (d, J= 4.03 Hz, 1 H), 7.58 (dd, J= 8.19, 1.34 Hz, 1 H), 7.33 (t, J= 8.19 Hz, 1 H), 7.23 (dd, J= 8.07, 1.34 Hz, 1 H), 3.53 - 3.43 (m, 1 H), 3.22 - 3.10 (m, 3 H), 3.09 (s, 3 H), 2.65 (s, 6 H), 1.93 (br d, J= 11.37 Hz, 1 H), 1.84 - 1.72 (m, 2 H), 1.53 (br s, 2 H), 1.42 - 1.26 (m, 2 H), 1.17 - 1.06 (m, 1 H), 1.02 - 0.91 (m, 1 H). HPLC: 97.44% (220 nm), 97.5% (215 nm), 97.53% (254 nm). MS (ESI): mass calcd. For C20H28CIN3O5S3521.09 m/z found 522.1 [M+H] ⁺ .

Example 134: N5-[3-chloro-2-[4-(methoxymethyl)-4-methyl-l-piperidyl] phenyl] -N2, N2- dimethyl-thiophene-2, 5-disulfonamide

[00627] Step 1. To a solution of tert-butyl 4-(hydroxymethyl)-4-methyl-piperidine-l -carboxylate (1 g, 4.36 mmol, 1 eq) in THF (10 mL) was added dropwise NaH (349 mg, 8.72 mmol, 60% purity, 2 eq) at 0 °C. After addition, the mixture was stirred at this temperature for 0.5 hour, and then Mel (1.86 g, 13.0 mmol, 814 μL, 3 eq) was added dropwise at 0 °C. The resulting mixture was stirred at 20 °C for 2 hours. LC-MS showed tert-butyl 4-(hydroxymethyl)-4-methyl-piperidine-l -carboxylate was consumed completely and one main peak with desired mass was detected. Then it was separated between H ₂ O (20 mL) and 4 EtOAc (40 mL). The organic phase was separated, washed with 30 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired tertbutyl 4-(methoxymethyl)-4-methyl-piperidine-l -carboxylate (1.2 g, crude) as ayellow oil. MS (ESI): mass calcd. ForCisEEsNOs 243.18 m/z found 188.2 [M+H] ⁺ . [00628] Step 2.' A mixture of tert-butyl 4-(methoxymethyl)-4-methyl-piperidine-l -carboxylate (1.2 g, 4.93 mmol, 1 eq) in HCl/EtOAc (4 M, 12 mL, 9.73 eq) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 1 hour under N2 atmosphere. TLC (petroleum ether: EtOAc = 3:1) indicated tert-butyl 4-(methoxymethyl)-4-methyl-piperidine-l -carboxylate was consumed completely and one new spot formed. The reaction mixture was concentrated in vacuum to give desired 4-(methoxymethyl)-4-methyl-piperidine (1 g, crude, HC1) as a yellow solid.

[00629] Step 3. A mixture of 4-(methoxymethyl)-4-methyl-piperidine (0.5 g, 3.49 mmol, 1 eq), 1 -chloro-2-fluoro-3 -nitro-benzene (613 mg, 3.49 mmol, 1 eq), DIEA (902 mg, 6.98 mmol, 1.22 mL,

2 eq) and K ₂ CO ₃ (965 mg, 6.98 mmol, 2 eq) in dioxane (5 mL) was degassed and purged with N2 for

3 times, and then the mixture was stirred at 120 °C for 12 hours under N2 atmosphere. LC-MS showed 4-(methoxymethyl)-4-methyl-piperidine (0.5 g, 3.49 mmol, 1 eq), 1 -chloro-2-fluoro-3 -nitrobenzene (613 mg, 3.49 mmol, 1 eq), DIEA (902 mg, 6.98 mmol, 1.22 mL, 2 eq) and K ₂ CO ₃ (965 mg, 6.98 mmol, 2 eq) in dioxane (5 mL) was consumed completely and one main peak with desired mass was detected. Then it was separated between H ₂ O (20 mL) and 40 mL of EtOAc. The organic phase was separated, washed with 30 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0~8% EtOAc /petroleum ether gradient @ 60 mL/min) to give desired l-(2-chloro-6-nitro-phenyl)-4-(methoxymethyl)-4- methyl-piperidine (960 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C14H19CIN2O3 298.11 m/z found 299.2 [M+H] ⁺ .

[00630] Step 4. To a solution of 1 -(2-chloro-6-nitro-phenyl)-4-(methoxymethy l)-4-methyl- piperidine (960 mg, 3.21 mmol, 1 eq) in EtOH (10 mL) and H ₂ O (2 mL) was added AcOH (772 mg, 12.9 mmol, 735 μL, 4 eq) and Fe (897 mg, 16.0 mmol, 5 eq). The mixture was stirred at 80 °C for 0.5 hour. LC-MS showed l-(2-chloro-6-nitro-phenyl)-4-(methoxymethyl)-4-methyl-piper idine was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove EtOH, then it was separated between H ₂ O (20 mL) and EtOAc (40 mL). The organic phase was separated, washed with 30 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 3-chloro-2-[4- (methoxymethyl)-4-methyl-l -piperidyl] aniline (1 g, crude) as a yellow solid. MS (ESI): mass calcd. For C14H21CIN2O 268.13 m/z found 269.2 [M+H] ⁺ .

[00631] Step 5: A mixture of 3 -chloro-2-[4-(methoxymethyl)-4-methyl-l -piperidyl] aniline (0.2 g, 690 pmol, 1 eq) and compound core 1 (371.01 mg, 1.38 mmol, 2 eq) in Py (3 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20 °C for 1 hour under N2 atmosphere. LC-MS showed 3 -chloro-2-[4-(methoxymethyl)-4-methyl-l -piperidyl] aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated in vacuum to give the crude product. The residue was purified by prep-WY,C (basic condition; according to LCMS; Method: column: Waters Xbridge Prep OBD C18 150*40mm*10um; mobile phase: [water (0.05%NH3H20+10mM NEEHCO3)-ACN]; B%: 40%-80%,8 min) to give desired N5-[3-chloro-2-[4-(methoxymethyl)-4-methyl-l-piperidyl]pheny l]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide (77.4 mg, 141 pmol, 20.4% yield, 95.0% purity) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.77 - 9.32 (m, 1H), 7.67 (s, 2H), 7.44 - 7.04 (m, 3H), 3.40 - 3.32 (m, 4H), 3.28 (br s, 3H), 3.21 - 3.07 (m, 2H), 2.64 (s, 6H), 1.57 (br d, J=10.3 Hz, 2H), 1.44 - 1.19 (m, 2H), 0.96 (s, 3H). HPLC: 94.96% (220 nm), 92.8% (215 nm), 96.68% (254 nm). MS (ESI): mass calcd. For C20H28CIN3O5S3 521.09 m/z found 522.0 [M+H] ⁺ .

Example 135: N5-[2-(3-azabicyclo [3.1.0] hexan-3-yl)-3-chloro-phenyl]-N2, N2-dimethyl- thiophene-2, 5-disulfonamide

[00632] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (300 mg, 1.71 mmol, 1 eq) and 3-azabicyclo [3.1.0] hexane hydrochloride (245 mg, 2.05 mmol, 1.2 eq) in dioxane (3 mL) was added K ₂ CO ₃ (709 mg, 5.13 mmol, 3 eq) and DIEA (663 mg, 5.13 mmol, 893 μL, 3 eq). The mixture was stirred at 120 °C for 12 hours. LC-MS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (10 mL) and extracted with EtOAc (5 mL * 3). The combined organic layers were washed with brine (5mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0-15% EtOAc/petroleum ether gradient @ 35 mL/min) to give desired 3 -(2-chloro-6-nitro-phenyl)-3 -azabicyclo [3.1.0] hexane (250 mg, 1.05 mmol, 61.3% yield) as ayellow oil. MS (ESI): mass calcd. For C11H11CIN2O2 238.05 m/z found 239.0 [M+l] ⁺ .

[00633] Step 2: To a solution of 3 -(2-chloro-6-nitro-phenyl)-3 -azabicyclo [3.1.0] hexane (250 mg, 1.05 mmol, 1 eq) in EtOH (3 mL) and H ₂ O (0.6 mL) was added Fe (585 mg, 10.5 mmol, 10 eq) and NH4CI (560 mg, 10.5 mmol, 10 eq). The mixture was stirred at 80 °C for 1 hour. LC-MS showed 3 -(2-chloro-6-nitro-phenyl)-3 -azabicyclo [3.1.0] hexane was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with H ₂ O (5 mL) and extracted with EtOAc (5 mL * 2). The combined organic layers were washed with brine (5 mL * 2), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 3/1) to give 2-(3 -azabicyclo [3.1.0] hexan-3-yl)-3 -chloro-aniline (140 mg, 670 umol, 64.1% yield) as ayellow oil. MS (ESI): mass calcd. For C11H13CIN2 208.08 m/z found 209.0 [M+l] ⁺ .

[00634] Step 3. To a solution of 2-(3 -azabicyclo [3.1.0] hexan-3-yl)-3 -chloro-aniline (140 mg, 671 pmol, 1 eq) and 5 -(dimethylsulfamoyl) thiophene-2-sulfonyl chloride (253mg, 872 pmol, 1.3 eq) in Py (2 mL). The mixture was stirred at 20 °C for 2 hours. LC-MS showed 2-(3 -azabicyclo [3.1.0] hexan-3-yl)-3 -chloro-aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove pyridine and get a residue. The residue was purified by prep-W ,C (neutral condition: column: Waters Xbridge BEH C18 100*25mm*5um; mobile phase: [water (10mM NHIHCO3)-ACN]; B%: 40%-75%,10min) to give desired N5-[2-(3-azabicyclo [3.1.0] hexan-3-yl)-3-chloro-phenyl]-N2, N2-dimethyl-thiophene- 2, 5 -disulfonamide (80 mg, 167 pmol, 24.9% yield, 96.6% purity) as a white solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 9.70 (s, 1H), 7.67 (d, J= 4.0 Hz, 1H), 7.59 (d, J= 4.0 Hz, 1H), 7.33 - 7.28 (m, 1H), 7.21 - 7.15 (m, 2H), 3.43 (br d, J= 7.6 Hz, 2H), 2.72 - 2.66 (m, 8H), 1.52 - 1.45 (m, 2H), 0.77 (q, J= 4.0 Hz, 1H), 0.50 (dt, J= 4.1, 7.7 Hz, 1H). HPLC: 96.59% (220 nm), 97.16% (215 nm), 97.35% (254 nm). MS (ESI): mass calcd. For C17H20CIN3O4S3 461.03 m/z found 462.1[[M+1] ⁺ .

Example 136: N-[3-chloro-2-(4, 4-dimethyl-l-piperidyl) phenyl] -5-cyclopro pylsulfonyl- thiophene-2-sulfonamide [00635] Step 1. To a solution of thiophene-2-thiol (2 g, 17.2 mmol, 1 eq) and KI (3.43 g, 20.7 mmol, 1.2 cc/)in DMSO (20 mL) was added dropwise t-BuOK (2.32 g, 20.7 mmol, 1.2 eq) at 0 °C. After addition, the mixture was stirred at this temperature for 0.5 hour, and then iodocyclopropane (2.89 g, 17.2 mmol, 1 eq) was added dropwise at 20 °C. The resulting mixture was stirred at 80 °C for 11.5 hours. TLC (petroleum ether/EtOAc = 10/1, Rf = 0.6) indicated thiophene-2-thiol was consumed and desired mass was detected. The crude was added H ₂ O (40 mL), and extracted with EtOAc (30 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 2-cyclopropylsulfanylthiophene (3.74 g, crude) as a brown oil.

[00636] Step 2: To a solution of 2-cyclopropylsulfanylthiophene (3.74 g, 24.0 mmol, 1 eq) in DCM (50 mL) was added dropwise m-CPBA (17.0 g, 83.8 mmol, 85% purity, 3.5 eq) at 0 °C. The resulting mixture was stirred at 15 °C for 12 hours. TLC (petroleum ether/EtOAc = 3/1, Rf = 0.3) indicated 2-cyclopropylsulfanylthiophene was consumed and desired mass was detected. The crude was added H ₂ O (100 mL), and extracted with EtOAc (50 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by chromatography (SiO2, petroleum ether/EtOAc = 3/1 to 4/1) to give desired 2-cyclopropylsulfonylthiophene (1.82 g, 9.67 mmol, 40.4% yield) as a brown oil.

[00637] Step 3. To a solution of 2-cyclopropylsulfonylthiophene (1 g, 5.31 mmol, 1 eq) in Br2 (849 mg, 5.31 mmol, 274 μL, 1 eq) was added AcOH (319 mg, 5.31 mmol, 304 μL, 1 eq) and NBS (945 mg, 5.31 mmol, 1 eq) and AC2O (2.17 g, 21.3 mmol, 1.99 mL, 4 eq). The mixture was stirred at 15 °C for 3 hours. TLC (petroleum ether/EtOAc = 3/1, Rf = 0.5) indicated 2- cyclopropylsulfonylthiophene was consumed and desired mass was detected. The crude was added H ₂ O (20 mL), and extracted with EtOAc (15 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by chromatography (SiO2, petroleum ether/EtOAc = 10/1 to 5/1) to give desired 2-bromo-5- cyclopropylsulfonyl-thiophene (0.9 g, 3.37 mmol, 63.4% yield) as a ping solid.

[00638] Step 4. A mixture of 2-bromo-5-cyclopropylsulfonyl-thiophene (500 mg, 1.87 mmol, 1 eq), phenylmethanethiol (279 mg, 2.25 mmol, 263 μL, 1.2 eq), Xantphos (108 mg, 187 pmol, 0.1 eq), DIEA (484 mg, 3.74 mmol, 652 μL, 2 eq) and Pd (dppf) CI2 (34.2 mg, 46.8 pmol, 0.025 eq) in Tol. (5 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 110 °C for 12 hours under N2 atmosphere. TLC (petroleum ether/EtOAc = 3/1, Rf = 0.5) indicated 2-bromo-5- cyclopropylsulfonyl-thiophene was consumed and desired mass was detected. The crude was added H ₂ O (40 mL), and extracted with EtOAc (30 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by chromatography (SiCL, petroleum ether/EtOAc = 5/1 to 4/1) to give desired 2-benzylsulfanyl-5- cyclopropylsulfonyl-thiophene (125 mg, 403 pmol, 21.5% yield) as a brown oil.

[00639] Step 5: To a solution of 2-benzylsulfanyl-5-cyclopropylsulfonyl-thiophene (125 mg, 403 pmol, 1 eq) in AcOH (1 mL) and H ₂ O (0.25 mL) was added NCS (161 mg, 1.21 mmol, 3 eq). The mixture was stirred at 20 °C for 2 hours. TLC (petroleum ether/EtOAc = 3/1, Rf = 0.3) indicated2- benzylsulfanyl-5-cyclopropylsulfonyl-thiophene was consumed and desired mass was detected. The crude was added H ₂ O (10 mL), and extracted with EtOAc 30 mL (10 mL * 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 5-cyclopropylsulfonylthiophene-2-sulfonyl chloride (115 mg, crude) as a brown oil.

[00640] Step 6: To a solution of 3-chloro-2-(4, 4-dimethyl-l -piperidyl) aniline (95.7 mg, 401 pmol, 1 eq) in Py (1 mL) was added 5-cyclopropylsulfonylthiophene-2-sulfonyl chloride (115 mg, 401 pmol, 1 eq). The mixture was stirred at 20 °C for 2 hours. LCMS showed 3-chloro-2-(4, 4- dimethyl-1 -piperidyl) aniline was consumed completely and desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 150*30mm*5um; mobile phase: [water (0.2%FA)- ACN]; B%: 45%-85%, 8min). The purified product was dissolved in H2O( 10 mL), and then the solution was lyophilized to give desired N-[3-chloro-2-(4,4-dimethyl-l-piperidyl)phenyl]-5- cyclopropylsulfonyl-thiophene-2-sulfonamide (15.1 mg, 30.6 pmol, 7.64% yield, 99.2% purity) as a faint yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 9.64 - 9.60 (m, 1 H), 7.89 - 7.85 (m, 1 H), 7.74 - 7.70 (m, 1 H), 7.37 - 7.32 (m, 2 H), 7.23 - 7.29 (m, 1 H), 3.37 - 3.29 (m, 4 H), 3.15 - 3.10 (m, 1 H), 1.58 - 1.47 (m, 2 H), 1.40 - 1.30 (m, 2 H), 1.23 - 1.18 (m, 4 H), 1.04 - 1.00 (m, 6 H). HPLC: 99.23% (220 nm), 99.25% (215 nm), 99.59% (254 nm). MS (ESI): mass calcd. For C20H25CIN2O4S3 488.07 m/z found 489.0 [M+H] ⁺ .

Example 137: N5-[2-(l, 3, 3a, 4, 6, 6a-hexahydrofuro [3, 4-c] pyrrol-5-yl)-3-chloro-phenyl]-N2,

N2-dimethyl-thiophene-2, 5-disulfonamide [00641] Step 1. To a solution of 1 -chloro-2-fluoro-3 -nitro-benzene (235 mg, 1.34 mmol, 1 eq) in dioxane (4 mL) was added K ₂ CO ₃ (370 mg, 2.68 mmol, 2 eq), 3, 3a, 4, 5, 6, 6a-hexahydro-lH- furo[3, 4-c]pyrrole (240 mg, 1.61 mmol, 1.2 eq, HC1) and DIEA (346 mg, 2.68 mmol, 466 μL, 2 eq). The mixture was stirred at 120 °C for 12 hours. LC-MS showed 1 -chloro-2-fluoro-3 -nitro-benzene was consumed completely and one main peak with desired mass was detected. The reaction mixture was diluted with water 30 mL and extracted with EtOAc 90 mL (30 mL * 3). The combined organic layers were washed with brine 90 mL (30 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give desired 5-(2-chloro-6-nitro-phenyl)-l, 3, 3a, 4, 6, 6a- hexahydrofuro[3, 4-c]pyrrole (450 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C12H13CIN2O3 268.06 m/z found 269.2 [M+H] ⁺ .

[00642] Step 2: To a solution of 5-(2-chloro-6-nitro-phenyl)-l, 3, 3a, 4, 6, 6a-hexahydrofuro [3, 4-c] pyrrole (200 mg, 744 pmol, 1 eq) in H ₂ O (0.5 mL) and EtOH (2 mL) was added Fe (208 mg, 3.72 mmol, 5 eq) and NH ₄ CI (398 mg, 7.44 mmol, 10 eq). The mixture was stirred at 80 °C for 0.5 hour. LC-MS showed 5-(2-chloro-6-nitro-phenyl)-l, 3, 3a, 4, 6, 6a-hexahydrofuro [3, 4-c] pyrrole was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was diluted with water 30 mL and extracted with EtOAc 90 mL (30 mL * 3). The combined organic layers were washed with brine 90 mL (30 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether/EtOAc = 3/1) to give desired 2-(l, 3, 3a, 4, 6, 6a-hexahydrofuro [3, 4-c] pyrrol-5-yl)-3-chloro- aniline (90 mg, 377 pmol, 50.7% yield) as a faint yellow oil. MS (ESI): mass calcd. For C12H15CIN2O 238.09 m/z found 239.2 [M+H] ⁺ .

[00643] Step 3. To a solution of 2-(l, 3, 3a, 4, 6, 6a-hexahydrofuro [3, 4-c] pyrrol-5-yl)-3-chloro- aniline (90 mg, 377 pmol, 1 eq) in Py (1 mL) was added 5-(dimethylsulfamoyl)thiophene-2-sulfonyl chloride (109 mg, 377 pmol, 1 eq). The mixture was stirred at 15 °C for 1 hour. LC-MS showed 2- (1, 3, 3a, 4, 6, 6a-hexahydrofuro [3, 4-c] pyrrol-5-yl)-3-chloro-aniline was consumed completely and one main peak with desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WY,C (basic condition; column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water (0.05%NH3H20+10mM NEEHCO3)-ACN]; B%: 35%-65%, 8min) to give desired N5-[2-(l, 3, 3a, 4, 6, 6a-hexahydrofuro[3, 4-c]pyrrol-5-yl)-3-chloro-phenyl]-N2, N2-dimethyl-thiophene-2, 5 -disulfonamide (83.8 mg, 170 pmol, 45.0% yield, 99.6% purity) as a yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ ,) δ 9.60 (s, 1 H), 7.71 - 7.67 (m, 1 H), 7.35 - 7.34 (m, 1 H), 7.34 (d, J= 2.13 Hz, 1 H), 7.28 - 7.20 (m, 2 H), 3.76 (br dd, J= 8.76, 6.38 Hz, 2 H), 3.62 (dd, J= 8.88, 2.63 Hz, 2 H), 3.28 (br dd, J= 8.44, 6.82 Hz, 2 H), 2.88 (br d, J= 2.50 Hz, 2 H), 2.74 (dd, J= 8.76, 1.88 Hz, 2 H), 2.67 (s, 6 H). HPLC: 99.60% (220 nm), 99.45% (215 nm), 99.86% (254 nm). MS (ESI): mass calcd. For C18H22CIN3O5S3491.04 m/z found 492.1 [M+H] ⁺ .

Example 138: [3-chloro-2-(4, 4-dimethyl-l-piperidyl)-N-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonyl] anilino] methyl dihydrogen phosphate

[00644] Step 1. To a solution of N5-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl]-N2, N2- dimethyl-thiophene-2, 5 -disulfonamide (300 mg, 610 pmol, 1 eq) in THF (2 mL) was added NaH (122 mg, 3.05 mmol, 60% purity, 5 eq) and H (7.74 mg, 30.5 pmol, 6.14 μL, 0.05 eq) at 0 °C. chloro(methylsulfanyl)methane (118 mg, 1.22 mmol, 102 μL, 2 eq) was added to the mixture at 0 °C. The mixture was stirred at 20 °C for 2 hours. LC-MS showed N5-[3-chloro-2-(4, 4-dimethyl-l- piperidyl) phenyl] -N2, N2-dimethyl-thiophene-2, 5 -disulfonamide remained and desired compound was detected. The crude was added H ₂ O (100 mL), and extracted with EtOAc (100 mL * 3). The combined organic layers were washed with brine (100 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 2/1) to give desired N2-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl]- N5, N5-dimethyl-N2-(methylsulfanylmethyl) thiophene-2, 5 -disulfonamide (330 mg, crude) as a yellow oil. MS (ESI): mass calcd. For C21H30CIN3O4S4551.08 m/z found 552.0 [M+H] ⁺ .

[00645] Step 2.' To a solution of N2-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) phenyl]-N5, N5- dimethyl-N2-(methylsulfanylmethyl) thiophene-2, 5-disulfonamide (330 mg, 598 pmol, 1 eq) in THF (5 mL) was added H3PO4 (586 mg, 5.98 mmol, 349 μL, 10 eq) and NIS (202 mg, 896 pmol, 1.5 eq). The mixture was stirred at 20 °C for 1 hour. LC-MS showed N2-[3-chloro-2-(4, 4-dimethyl-l- piperidyl) phenyl] -N5, N5-dimethyl-N2-(methylsulfanylmethyl) thiophene-2, 5-disulfonamide was consumed completely and desired mass was detected. The reaction mixture was added to sat. aq. Na ₂ SO ₃ (10 mL), then the mixture was concentrated under reduced pressure to remove THF. The reaction mixture was added to sat. aq. NaHCO3(20 mL), extracted with EtOAc (10 mL*3). The combined organic layers were washed with brine (20 mL) and dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-WY,C (neutral condition; column: Waters Xbridge Prep OBD C18 150*40mm* l Oum; mobile phase: [water (10mM NH4HCOS)-ACN]; B%: 30%-60%, 8min) to give desired [3-chloro-2-(4, 4-dimethyl-l- piperidyl)-N-[[5-(dimethylsulfamoyl)-2-thienyl]sulfonyl]anil ino]methyl dihydrogen phosphate (15 mg, 24.0 pmol, 4.01% yield, 93.7% purity) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ,) δ 8.09 (d, J= 4.0 Hz, 1H), 7.74 (d, J= 4.1 Hz, 1H), 7.46 (dd, J= 1.4, 7.9 Hz, 1H), 7.02 - 6.88 (m, 2H), 5.72 (dd, J= 4.8, 10.3 Hz, 1H), 4.82 (dd, J= 4.1, 10.3 Hz, 1H), 3.13 (br d, J= 12.5 Hz, 4H), 2.72 (s, 6H), 1.41 (br s, 4H), 0.99 (s, 6H). HPLC: 88.60% (220 nm), 83.38% (215 nm), 93.69% (254 nm). MS (ESI): mass calcd. For C20H29CIN3O8PS3601.05 m/z found 602.1 [M+H] ⁺ .

Example 139: 3-[3-chloro-2-(4, 4-dimethyl-l-piperidyl)-N-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonyl] anilino] propanoic acid

[00646] Step 1. To a solution of 3-chloro-2-(4, 4-dimethyl-l-piperidyl)aniline (1 g, 4.19 mmol, 1 eq) in tert-butyl prop-2-enoate (1.07 g, 8.38 mmol, 1.22 mL, 2 eq) was added imidazole (131 mg, 1.26 mmol, 0.3 eq, HC1). The mixture was stirred at 110 °C for 12 hours. LC-MS showed 3-chloro-

2-(4, 4-dimethyl-l -piperidyl) aniline remained and one main peak with desired mass was detected. The reaction mixture was diluted with water 50 mL and extracted with EtOAc 150 mL (50 mL * 3). The combined organic layers were washed with brine 150 mL (50 mL * 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiCL, petroleum ether/Ethyl acetate=l/O to 2/1) to give desired tert-butyl

3-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) anilino] propanoate (1 g, 2.73 mmol, 65.1% yield) as a yellow oil. MS (ESI): mass calcd. For C20H31CIN2O2 366.21 m/z found 367.3 [M+H] ⁺ .

[00647] Step 2\ To a solution of tert-butyl 3-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) anilino] propanoate (150 mg, 409 pmol, 1 eq) in Py (3 mL) was added 5 -(dimethylsulfamoyl) thiophene-2- sulfonyl chloride (261 mg, 899 pmol, 2.2 eq). The mixture was stirred at 40 °C for 12 hours. LC-MS showed tert-butyl 3-[3-chloro-2-(4, 4-dimethyl-l -piperidyl) anilino] propanoate remained and desired compound was detected. The reaction mixture was concentrated to give the crude product. The residue was purified by prep-TLC (SiCL, petroleum ether/EtOAc = 3/1) to give desired tert-butyl 3- [3-chloro-2-(4, 4-dimethyl-l-piperidyl)-N-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonyl] anilino] propanoate (210 mg, 339 pmol, 82.8% yield) as an orange oil. MS (ESI): mass calcd. For C26H38CIN3O6S3 619.16 m/z found 620.3 [M+H] ⁺ .

[00648] Step 3: To a solution of tert-butyl 3-[3-chloro-2-(4, 4-dimethyl-l-piperidyl)-N-[[5- (dimethylsulfamoyl)-2-thienyl] sulfonyl] anilino] propanoate (200 mg, 322 pmol, 1 eq) in TFA (5 mL). The mixture was stirred at 25 °C for 1 hour. LC-MS showed tert-butyl 3-[3-chloro-2-(4, 4- dimethyl-l-piperidyl)-N-[[5-(dimethylsulfamoyl)-2-thienyl] sulfonyl] anilino] propanoate was consumed completely and one main peak with desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove THF to give a residue. The residue was purified by prep-HPLC (neutral condition) to give desired 3-[3-chloro-2-(4, 4-dimethyl-l-piperidyl)-N-[[5- (dimethylsulfamoyl)-2-thienyl] sulfonyl] anilino] propanoic acid (70.1 mg, 124 pmol, 38.3% yield, 99.4% purity) as a white solid. ¹ H NMR (400MHz, DMSO-d6 ) δ 12.34 (br s, 1H), 7.84 (d, ./ =4.0 Hz, 1H), 7.76 (d, J= 4.0 Hz, 1H), 7.49 (dd, J= 1.4, 8.0 Hz, 1H), 7.17 (t, J = 8.0 Hz, 1H), 6.98 (dd, J = 1.4, 7.9 Hz, 1H), 4.07 - 3.76 (m, 2H), 3.56 - 3.39 (m, 2H), 2.91 (br d, J= 5.6 Hz, 2H), 2.74 (s, 6H), 2.47 - 2.29 (m, 2H), 1.42 (br s, 4H), 0.99 (s, 6H). HPLC: 99.40% (220 nm), 99.33% (215 nm), 99.85% (254 nm). MS (ESI): mass calcd. For C22H30CIN3O6S3 563.10 m/z found 564.1 [M+H] ⁺ .

Biological Assays

Mammalian whole-cell electrophysiology.

[00649] HEK293T, Cos-1 and CHO cells were used for the heterologous expression experiments and were transfected using Lipofectamine 2000 (Invitrogen). The pipette solution contained 147 mM Cs, 120 mM methane-sulphonate, 4 mM NaCl, 10 mM EGTA, 2 mM Na2-ATP, 2 mM MgCh and 20 mM HEPES (pH 7.2; free [Ca2 + ]i < 10 nM). The standard extracellular bath solution (modified Tyrode’s solution) contained 153 mM NaCl, 5 mM KC1, 2 mM CaCh, 1 mM MgCh, 20 mM HEPES and 10 mM glucose (pH 7.4). The ‘Low pH Tyrode’ solution contained 150 mM Na-gluconate, 5 mM KC1, 2 mM CaCl ₂ , 1 mM MgCh, 10 mM glucose, 10 mM HEPES and 10 mM MES (pH 4.6). All bath solutions were applied via a perfusion system to achieve a complete solution exchange within a few seconds. Data were collected using an Axopatch 2A patch clamp amplifier, Digidata 1440 and pClamp 10.0 software (Axon Instruments). Whole-cell currents were digitized at 10 kHz and filtered at 2 kHz. All experiments were conducted at room temperature (21-23 °C), and all recordings were analysed with pClamp 10.0 and Origin 8.0 (OriginLab, Northampton, MA, USA).

Endolysosomal electrophysiology. [00650] Endolysosomal electrophysiology was performed in isolated endolysosomes using a modified patch-clamp method. Briefly, cells were treated with 1 μM vacuolin-1 for 2-5 h to increase the size of endosomes and lysosomes. Whole-endolysosome recordings were performed on isolated enlarged LELs. The bath (intemal/cytoplasmic) solution contained 140 mM K-gluconate, 4 mM NaCl, 1 mM EGTA, 2 mM Na2-ATP, 2 mM MgCl ₂ , 0.39 mM CaCl ₂ , 0.2 mM GTP and 10 mM HEPES (pH adjusted with KOH to 7.2; free [Ca ²⁺ ]i -100 nM based on the Maxchelator software (http://maxchelator.stanford.edu/)). The pipette (luminal) solution consisted of a ‘Low pH Tyrode’s solution with 145 mM NaCl, 5 mM KC1, 2 mM CaCl ₂ , 1 mM MgCl ₂ , 10 mM HEPES, 10 mM MES and 10 mM glucose (pH 4.6).

Lysosomal Ca ²⁺ imaging

[00651] GCaMP3-MLl expression was induced in Tet-On HEK-GCaMP3-MLl cells 20-24 h prior to experiments using 0.01 pg/mL doxycycline. GCaMP3-MLl fluorescence was monitored at an excitation wavelength of 470 nm (F470) using an EasyRatio Pro system (PTI). Cells were bathed in Tyrode’s solution containing 145 mM NaCl, 5 mM KC1, 2 mM CaCl ₂ , 1 mM MgCl ₂ , 10 mM Glucose, and 20 mM Hepes (pH 7.4). Lysosomal Ca ²⁺ release was measured in a zero Ca ²⁺ solution containing 145 mM NaCl, 5 mM KC1, 3 mM MgCl ₂ , 10 mM glucose, 1 mM EGTA, and 20 mM HEPES (pH 7.4). Ca ²⁺ concentration in the nominally free Ca ²⁺ solution is estimated to be 1-10 μM. With 1 mM EGTA, the free Ca ²⁺ concentration is estimated to be < 10 nM based on the Maxchelator software (http://maxchelator.stanford.edu/). Experiments were carried out 0.5 to 6 hrs after plating.

[00652] Exemplary EC50 data are provided in Table 1.

TFEB nuclear translocation assay

[00653] TFEB is a transcription factor and master regulator of lysosome biogenesis and autophagy. TFEB activation is shown to induce cellular clearance in a variety of LSDs and common neurodegenerative diseases. Hence TFEB activity can be used to evaluate the cellular efficacy of TRPML agonists.

[00654] Intracellular TFEB localization is determined either by immunofluorescence in Hela cells or by fluorescent microscopy in Hela cells stably expressing TFEB-GFP. Detailed procedures are as follows.

[00655] Cells grown on cover slips were treated with TRPML 1 agonist(s) or antagonist(s) for indicated time period and then fixed with 4% paraformaldehyde for 15 minutes at room temperature. For immunofluorescent detection of endogenous TFEB, cells were permeabilized with 0.3% Triton X-100, blocked with 1% bovine serum albumin (BSA) in phosphate buffered saline (PBS) and then immunostained with anti-TFEB antibody (1:200; Cell Signaling Technology) at 4°C for overnight. The stained cells were then incubated with secondary antibodies conjugated to Alexa Fluor 568 or 488 (ThermoFisher) for 1 h, then 4',6-diamidino-2-phenylindole (DAPI) for 15 min (to stain the cell nucleus) and mounted on glass slides with Fluoromount-G (Southern Biotech) for observation. Images were acquired with a Spinning-Disk Confocal microscope (Olympus) and Metamorph software (Molecular Devices).

[00656] Average ratios of nuclear versus cytosolic TFEB fluorescence intensity (>50 randomly - selected cells per experiment) were determined with ImageJ software (NIH).

[00657] Exemplary data of TFEB nuclear translocation assay are provided in Table 1.

UV treatment and ROS staining

[00658] HaCat cells (a human keratinocyte cell line) at a confluency of around 50% were incubated with 20 μM H2DCFDA (Sigma) in the culture media without FBS at 37 °C for 30 min, and then washed twice with phosphate buffered saline (PBS, pH 7.4) before being exposed to UV. A CL- 1000 UV Crosslinker (UVP) is used to shed 600 J/m ² of UV light (254 nm) on the HaCat cells in all experiments. Fluorescent images were obtained right after UV exposure, via an Olympus 1X73 microscopy (Olympus) and Metamorph software (Molecular Devices).

[00659] Applicant’s disclosure is described herein in preferred embodiments with reference to the Figures, in which like numbers represent the same or similar elements. Reference throughout this specification to “one embodiment,” “an embodiment,” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases “in one embodiment,” “in an embodiment,” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

[00660] The described features, structures, or characteristics of Applicant’s disclosure may be combined in any suitable manner in one or more embodiments. In the description, herein, numerous specific details are recited to provide a thorough understanding of embodiments of the invention.

One skilled in the relevant art will recognize, however, that Applicant’s composition and/or method may be practiced without one or more of the specific details, or with other methods, components, materials, and so forth. In other instances, well-known structures, materials, or operations are not shown or described in detail to avoid obscuring aspects of the disclosure.

[00661] Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Methods recited herein may be carried out in any order that is logically possible, in addition to a particular order disclosed.

Incorporation by Reference

[00662] References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made in this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes. Any material, or portion thereof, that is said to be incorporated by reference herein, but which conflicts with existing definitions, statements, or other disclosure material explicitly set forth herein is only incorporated to the extent that no conflict arises between that incorporated material and the present disclosure material. In the event of a conflict, the conflict is to be resolved in favor of the present disclosure as the preferred disclosure.

Equivalents

[00663] The representative examples are intended to help illustrate the invention, and are not intended to, nor should they be construed to, limit the scope of the invention. Indeed, various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including the examples and the references to the scientific and patent literature included herein. The examples contain important additional information, exemplification and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.