CARBONYL LINKED BICYCLIC HETEROARYL ANTIBIOTIC TOLERANCE INHIBITORS

TECHNICAL FIELD

[0001] The present disclosure includes carbonyl linked bicyclic heteroaryl compounds and related compounds and methods of using such compounds. For example, the disclosure provides methods of using the compounds in treating acute and chronic bacterial infections.

BACKGROUND

[0002] Hard-to-eradicate, often unbeatable, infections including chronic wounds and infections of medical devices pose increasing threats to human health worldwide. Such infections are often refractory to antibiotics due to antibiotic resistant bacterial cells, and/or to antibiotic tolerance of a subpopulation of bacterial cells that are not antibiotic resistant mutants but rather "dormant" cells that survive antibiotic killing. Antibiotic tolerance is defined as the ability of a fraction of a susceptible bacterial population to survive exposure to normally lethal concentrations of bactericidal antibiotics. According to the existing paradigm, many chronic infections are therefore unbeatable.

[0003] The ligand activated transcriptional regulator, MvfR, plays a central role in controlling the pathology of acute bacterial infections, and the shift of Gram-negative bacteria from acute to chronic infection. MvfR inhibitors reduce virulence of Pseudomonas aeruginosa, a Gram-negative bacterial species, and reduce the formation of antibiotic resistant Pseudomonas strains in vitro.

SUMMARY

[0004] In a first embodiment, this disclosure includes compounds of Formula (I) and the pharmaceutically acceptable salts, hydrates, solvates and prodrugs thereof.

[0005] Q is selected from:

[0006] Each of Y ¹ , Y ² , Y ³ , Y ⁴ , Y ⁵ , Y ⁶ , Y ⁷ , Y ⁸ , and Y ⁹ is independently selected from N, CH, and

C(R ² ).

[0007] Each of Y ¹⁰ and Y ¹¹ is independently selected from N and C.

[0008] At least one of Y ¹ , Y ² , Y ³ , or Y ⁴ is C(R ² ).

[0009] No more than two of Y ¹ , Y ² , Y ³ , Y ⁴ , Y ¹⁰ and Y ¹¹ is N.

[0010] At least one of Y ⁶ , Y ⁷ , Y ⁸ , or Y ⁹ is C(R ² ).

[0011] No more than two of Y ⁶ , Y ⁷ , Y ⁸ , and Y ⁹ is N.

[0012] W is C and, when X is a bond or -C(R ^la )(R ^lb )-, W is additionally selected from N.

[0013] Z is selected from S, O, N, N(H) and NCd-Cs-alkyl), wherein the alkyl portion of Z, if present, is optionally substituted.

[0014] Each " " represents independently a single or a double bond, wherein at least one of the bonds between Y ¹⁰ and Y ⁵ or W and Y ⁵ is a double bond.

[0015] X is selected from a bond, -S-, -0-, -N(R ⁶ )-, and -C(R ^la )(R ^lb )-.

[0016] Each R ^la and each R ^lb is independently selected from hydrogen, halogen, -OH, -NH ₂ , -CN, -Ci-C6-alkyl, -C ₂ -C6 -alkenyl, -C ₂ -C6-alkynyl, -(Ci-C ₄ -alkylene)-(heterocycle), -(C ₂ -C -alkenylene)- (heterocycle), -(C ₂ -C ₄ -alkynylene)-heterocycle, -(Ci-C ₄ -alkylene)-(carbocycle), -(C ₂ -C -alkenylene)- (carbocycle), and -(C ₂ -C -alkynylene)-(carbocycle), or any R ^la and R ^lb bound to the same carbon atom are optionally taken together with the carbon atom to form an optionally substituted carbocycle or heterocycle.

[0017] Each R ² is independently selected from halogen, -OH, -NH ₂ , -CN, -Ci-C ₆ -alkyl, -C ₂ -C ₆ - alkenyl, -C ₂ -C6-alkynyl, -(C ₀ -C -alkylene)-(heterocycle), -(C ₂ -C -alkenylene) -(heterocycle), -(C ₂ -C - alkynylene) -heterocycle), -(C ₀ -C -alkylene)-(carbocycle), -(C ₂ -C -alkenylene)-(carbocycle), and -(C ₂ -C - alkynylene) -(carbocycle), or any two R ² are optionally taken together with the carbon atoms to which they are bound to form an optionally substituted carbocycle or heterocycle;

[0018] R ³ is selected from hydrogen, -Ci-C ₆ -alkyl, -C ₂ -C ₆ -alkenyl, -C ₂ -C ₆ -alkynyl, -(C ₀ -C ₄ - alkylene) -(heterocycle), -(C ₂ -C ₄ -alkenylene) -(heterocycle), -(C ₂ -C ₄ -alkynylene)-heterocycle), -(C ₀ -C ₄ - alkylene) -(carbocycle), -(C ₂ -C ₄ -alkenylene)-(carbocycle), and -(C ₂ -C ₄ -alkynylene)-(carbocycle). [0019] R ⁴ is selected from hydrogen, -CN, OH, NH ₂ , C0 ₂ H, -C(0)NH ₂ , -d-Cs-alkyl, -C ₂ -C ₆ - alkenyl, -C ₂ -C6-alkynyl, -(C ₀ -C ₄ -alkylene)-(heterocycle), -(C ₂ -C ₄ -alkenylene)-(heterocycle), -(C ₂ -C - alkynylene)-heterocycle), -(C ₀ -C ₄ -alkylene)-(carbocycle), -(C ₂ -C -alkenylene)-(carbocycle), and -(C ₂ -C - alkynylene) -(carbocycle) .

[0020] R ⁵ is selected from hydrogen, -CN, -OH, -NH ₂ , -C(0)NH ₂ , -d-C ₆ -alkyl, -C ₂ -C ₆ -alkenyl, -(C ₀ -C ₄ -alkylene)-(heterocycle), -(C ₂ -C ₄ -alkenylene)-(heterocycle), -(C ₀ -C ₄ -alkylene)-(carbocycle), and - (C ₂ -C ₄ -alkenylene)-(carbocycle).

[0021] Each R ⁶ is independently selected from hydrogen, Ci-C ₃ -alkyl, and Ci-C ₃ -fluoroalkyl.

[0022] Each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, heterocycle or carbocycle portion of any R ^la , R ^lb , R ² , R ³ , R ⁴ and R ⁵ is optionally and independently substituted.

[0023] One or more methylene units in the alkyl, alkenyl, alkynyl, alkylene, alkenylene, or alkynylene portion of any R ^la , R ^lb , R ² , R ³ , and R ⁴ and in the alkyl, alkenyl, alkylene, or alkenylene portion of R ⁵ are optionally and independently replaced with -0-, -S-, -N(R ⁶ )-, -S(=0)- or -S(=0) ₂ -.

[0025] The disclosure further includes a method of treating a bacterial infection in a subject, comprising administering a therapeutically effective amount of a compound, salt, solvate, or hydrate of Formula (I) or a prodrug thereof to a subject in need of such treatment. The compound of Formulae (I) may be administered as the only active agent or may be administered together with one or more additional active agents.

DETAILED DESCRIPTION

CHEMICAL DESCRIPTION AND TERMINOLOGY

[0026] Compounds are described using standard nomenclature. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this disclosure belongs. Unless clearly contraindicated by the context each compound name includes the free acid or free base form of the compound as well as all pharmaceutically acceptable salts, solvates, and hydrates of the compound.

[0027] The term "Formula (I)" encompasses all compounds that satisfy Formula (I), including any enantiomers, racemates and stereoisomers, as well as all pharmaceutically acceptable salts, solvates, and hydrates of such compounds. "Formula (I)" includes all subgeneric groups of Formula (I) unless clearly contraindicated by the context in which this phrase is used.

[0028] The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The term "or" means "and/or". The open-ended transitional phrase "comprising" encompasses the intermediate transitional phrase "consisting essentially of and the close-ended phrase "consisting of." Claims reciting one of these three transitional phrases, or with an alternate transitional phrase such as "containing" or "including" can be written with any other transitional phrase unless clearly precluded by the context or art. Recitation of ranges of values are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. The endpoints of all ranges are included within the range and independently combinable. All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as"), is intended merely for illustration and does not pose a limitation on the scope of the disclosure unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention as used herein. Unless defined otherwise, technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this disclosure belongs.

[0029] Compounds of Formula (I) include all compounds of Formula (I) having isotopic substitutions at any position. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example, and without limitation, isotopes of hydrogen include tritium and deuterium and isotopes of carbon include ⁿ C, ¹³ C, and ¹⁴ C.

[0030] An "active agent" means a compound (including a compound disclosed herein), element, or mixture that when administered to a subject, alone or in combination with another compound, element, or mixture, confers, directly or indirectly, a physiological effect on the subject. The indirect physiological effect may occur via a metabolite or other indirect mechanism.

[0031] A dash ("-") that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -C(0)NH ₂ is attached through carbon of the keto C(O) group.

[0032] An "aliphatic group" is a hydrocarbon group having the indicated number of carbon atoms in which the carbon atoms are covalently bound in single, double or triple covalent bonds in straight chains, branched chains, or non-aromatic rings. Aliphatic groups may be substituted.

[0033] "Alkyl" is a branched or straight chain saturated aliphatic hydrocarbon group, having the specified number of carbon atoms, generally from 1 to about 8 carbon atoms. The term Ci-C ₆ -alkyl as used herein indicates an alkyl group having from 1, 2, 3, 4, 5, or 6 carbon atoms. Other embodiments include alkyl groups having from 1 to 6 carbon atoms, 1 to 4 carbon atoms or 1 or 2 carbon atoms, e.g. Ci-C ₈ -alkyl, Ci-C ₄ -alkyl, and Ci-C ₂ -alkyl. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, t-butyl, n-pentyl, and sec-pentyl.

[0034] "Alkenyl" is a branched or straight chain aliphatic hydrocarbon group having one or more double carbon-carbon bonds that may occur at any stable point along the chain, having the specified number of carbon atoms. Examples of alkenyl include, but are not limited to, ethenyl and propenyl. [0035] "Alkynyl" is a branched or straight chain aliphatic hydrocarbon group having one or more triple carbon-carbon bonds that may occur at any stable point along the chain, having the specified number of carbon atoms. Examples of alkynyl include, but are not limited to, ethynyl and propynyl.

[0036] "Alkoxy" is an alkyl group as defined above with the indicated number of carbon atoms covalently bound to the group it substitutes by an oxygen bridge (-0-). Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, 2-butoxy, t-butoxy, n-pentoxy, 2-pentoxy, 3- pentoxy, isopentoxy, neopentoxy, n-hexoxy, 2-hexoxy, 3-hexoxy, and 3- methylpentoxy.

[0037] "Alkylthio" indicates an alkyl group as defined above attached through a sulfur linkage, i.e. a group of the formula alkyl-S-. Examples include ethylthio and pentylthio.

[0038] "Alkanoyl" is an alkyl group as defined above with the indicated number of carbon atoms covalently bound to the group it substitutes through a carbonyl C(O) bridge. The carbonyl carbon is included in the number of carbons, that is C ₂ alkanoyl is a CH ₃ C(0)- group.

[0039] "Alkylester" is an alkyl group as defined herein covalently bound to the group it substitutes by an ester linkage. The ester linkage may be in either orientation, e.g., a group of the formula -OC(0)-alkyl or a group of the formula -C(0)0-alkyl.

[0040] "Aryl" indicates aromatic groups containing only carbon in the aromatic ring or rings. Typical aryl groups contain 1 to 3 separate, fused, or pendant rings and from 6 to about 18 ring atoms, without heteroatoms as ring members. When indicated, such aryl groups may be further substituted with carbon or non-carbon atoms or groups. Aryl groups include, for example, phenyl, naphthyl, including 1- naphthyl, 2-naphthyl, and bi-phenyl.

[0041] A "carbocycle" is a monocyclic or bicyclic saturated, partially unsaturated, or aromatic ring system in which all ring atoms are carbon. Usually each ring of the carbocycle contains from 4-6 ring atoms and a bicyclic carbocycle contains from 7 to 10 ring atoms, but some other number of ring atoms may be specified. Unless otherwise indicated, the carbocycle may be attached to the group it substitutes at any carbon atom that results in a stable structure. When indicated the carbocyclic rings described herein may be substituted at any carbon atom if the resulting compound is stable. Examples of carbocycles include phenyl, naphthyl, tetrahydronaphthyl, cyclopropyl, cyclohexyl, and cyclohexenyl.

[0042] "Cycloalkyl" is a saturated hydrocarbon ring group, having the specified number of carbon atoms. Monocyclic cycloalkyl groups typically have from 3 to about 8 carbon ring atoms or from 3 to 6 (3, 4, 5, or 6) carbon ring atoms. Cycloalkyl substituents may be pendant from a substituted nitrogen, oxygen, or carbon atom, or a substituted carbon atom that may have two substituents may have a cycloalkyl group, which is attached as a spiro group. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0043] "Halo" or "halogen" indicates any of fluoro, chloro, bromo, and iodo. [0044] "Haloalkyl" indicates both branched and straight-chain alkyl groups having the specified number of carbon atoms, substituted with 1 or more halogen atoms, up to the maximum allowable number of halogen atoms. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and penta-fluoroethyl.

[0045] "Haloalkoxy" indicates a haloalkyl group as defined herein attached through an oxygen bridge (oxygen of an alcohol radical).

[0046] The term "heterocycle" indicates a monocyclic saturated, partially unsaturated, or aromatic ring containing from 1 to 4 heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon, or a bicyclic saturated, partially unsaturated, or aromatic heterocycle containing at least 1 heteroatom chosen from N, O, and S in one of the two rings of the two ring system and containing up to about 4 heteroatoms independently chosen from N, O, and S in each ring of the two ring system. Usually each ring of the heterocycle contains from 4-6 ring atoms but some other number of ring atoms may be specified. Unless otherwise indicated, the heterocycle may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. When indicated the heterocycles described herein may be substituted on carbon, sulfur, or nitrogen atom if the resulting compound is stable. It is preferred that the total number of heteroatoms in a heterocycle is not more than 4 and that the total number of S and O atoms in a heterocycle is not more than 2, more preferably not more than 1. Examples of heterocycles include, pyridyl, indolyl, pyrimidinyl, pyridazinyl, pyrazinyl, imidazolyl, oxazolyl, furanyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl,

benz[b]thiophenyl, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl, isoindolyl, dihydroisoindolyl, 5,6,7, 8-tetrahydroisoquinoline, pyrazolyl, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl. In certain embodiments a heterocycle is chosen from pyridinyl, pyrimidinyl, furanyl, thienyl, and pyrrolyl.

[0047] Additional examples of heterocycles include, but are not limited to, phthalazinyl, indolizinyl, indazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, benzoisoxolyl,

dihydro-benzodioxinyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, oxazolopyridinyl, imidazopyridinyl, isothiazolyl, naphthyridinyl, cinnolinyl, carbazolyl, beta-carbolinyl, isochromanyl, chromanonyl, chromanyl, tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl, pyridopyridinyl, benzotetrahydroiuranyl, benzotetrahydrothienyl, purinyl, benzodioxolyl, triazinyl, phenoxazinyl, phenothiazinyl, 5 pteridinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl, dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl, dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl, coumarinyl, isocoumarinyl, chromanyl, tetrahydroquinolinyl, dihydroquinolinyl, dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl,

dihydroisocoumarinyl, isoindolinonyl, benzodioxanyl, benzoxazolinonyl, pyrrolyl N-oxide, pyrimidinyl N-oxide, pyridazinyl N-oxide, pyrazinyl N-oxide, quinolinyl N-oxide, indolyl N-oxide, indolinyl N oxide, isoquinolyl N-oxide, quinazolinyl N-oxide, quinoxalinyl N-oxide, phthalazinyl N-oxide, imidazolyl N-oxide, isoxazolyl N-oxide, oxazolyl N- oxide, thiazolyl N-oxide, indolizinyl N oxide, indazolyl N-oxide, benzothiazolyl N-oxide, benzimidazolyl N-oxide, pyrrolyl N-oxide, oxadiazolyl N-oxide, thiadiazolyl N-oxide, tetrazolyl N- oxide, benzothiopyranyl S-oxide, and benzothiopyranyl S,S-dioxide.

[0048] "Heteroaryl" is a stable monocyclic aromatic ring having the indicated number of ring atoms which contains from 1 to 4, or in some embodiments from 1 to 2, heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon, or a stable bicyclic or tricyclic system containing at least one 5- to 7-membered aromatic ring which contains from 1 to 3, or in some embodiments from 1 to 2, heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon. Monocyclic heteroaryl groups typically have from 5 to 7 ring atoms. In some embodiments bicyclic heteroaryl groups are 9- to 10-membered heteroaryl groups, that is, groups containing 9 or 10 ring atoms in which one 5- to

7-member aromatic ring is fused to a second aromatic or non-aromatic ring. When the total number of S and O atoms in the heteroaryl group exceeds 1, these heteroatoms are not adjacent to one another. It is preferred that the total number of S and O atoms in the heteroaryl group is not more than 2. It is particularly preferred that the total number of S and O atoms in the aromatic heterocycle is not more than 1. Examples of heteroaryl groups include, but are not limited to, oxazolyl, pyranyl, pyrazinyl, pyrazolopyrimidinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinolinyl, tetrazolyl, thiazolyl, thienylpyrazolyl, thiophenyl, triazolyl, benzo[i ]oxazolyl, benzofuranyl, benzothiazolyl, benzothiophenyl, benzoxadiazolyl, dihydrobenzodioxynyl, furanyl, imidazolyl, indolyl, and isoxazolyl.

[0049] The term "mono- and/ or di-alkylamino" indicates secondary or tertiary alkyl amino groups, wherein the alkyl groups are independently chosen alkyl groups, as defined herein, having the indicated number of carbon atoms. The point of attachment of the alkylamino group is on the nitrogen. Examples of mono- and di-alkylamino groups include ethylamino, dimethylamino, and

methyl-propyl-amino.

[0050] The term "substituted", as used herein, means that any one or more hydrogens on the designated atom or group is replaced with a selection from the indicated group, provided that the designated atom's normal valence is not exceeded. When the substituent is oxo (i.e., =0) then 2 hydrogens on the atom are replaced. When an oxo group substitutes aromatic moieties, the corresponding partially unsaturated ring replaces the aromatic ring. For example a pyridyl group substituted by oxo is a pyridone. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure is meant to imply a compound that is sufficiently robust to survive isolation from a reaction mixture and subsequent formulation into an effective therapeutic agent. Unless otherwise specified substituents are named into the core structure. For example, it is to be understood that when aminoalkyl is means the point of attachment of this substituent to the core structure is in the alkyl portion and when alkylamino means the point of attachment is a bond to the nitrogen of the amino group.

[0051] Suitable groups that may be present on a "substituted" or "optionally substituted" position include, but are not limited to, e.g., halogen; cyano; -OH; -NH ₂ ; nitro; azido; alkanoyl (such as a C ₂ -C ₆ alkanoyl group); C(0)NH ₂ ; alkyl groups (including cycloalkyl groups) having 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms; alkenyl and alkynyl groups including groups having one or more unsaturated linkages and from 2 to about 8, or 2 to about 6 carbon atoms; alkoxy groups having one or more oxygen linkages and from 1 to about 8, or from 1 to about 6 carbon atoms; aryloxy such as phenoxy; alkylthio groups including those having one or more thioether linkages and from 1 to about 8 carbon atoms, or from 1 to about 6 carbon atoms; alkylsulfinyl groups including those having one or more sulfinyl linkages and from 1 to about 8 carbon atoms, or from 1 to about 6 carbon atoms; alkylsulfonyl groups including those having one or more sulfonyl linkages and from 1 to about 8 carbon atoms, or from 1 to about 6 carbon atoms; aminoalkyl groups including groups having one or more N atoms and from 1 to about 8, or from 1 to about 6 carbon atoms; aryl having 6 or more carbons and one or more rings, (e.g., phenyl, biphenyl, naphthyl, or the like, each ring either substituted or unsubstituted aromatic); arylalkyl having 1 to 3 separate or fused rings and from 6 to about 18 ring carbon atoms, with benzyl being an exemplary arylalkyl group; arylalkoxy having 1 to 3 separate or fused rings and from 6 to about 18 ring carbon atoms, with benzyloxy being an exemplary arylalkoxy group; or a saturated, unsaturated, or aromatic heterocycle having 1 to 3 separate or fused rings with 3 to about 8 members per ring and one or more N, O or S atoms, e.g. coumarinyl, quinolinyl, isoquinolinyl, quinazolinyl, pyridyl, pyrazinyl, pyrimidinyl, furanyl, pyrrolyl, thienyl, thiazolyl, triazinyl, oxazolyl, isoxazolyl, imidazolyl, indolyl, benzofuranyl, benzothiazolyl, tetrahydroiuranyl, tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, and pyrrolidinyl. Such heterocycles may be further substituted, e.g. with hydroxy, alkyl, alkoxy, halogen and amino. In certain embodiments "optionally substituted" includes one or more substituents independently chosen from halogen, hydroxyl, amino, cyano, -CHO, -C0 ₂ H, -C(0)NH ₂ , Ci-C ₆ -alkyl, C ₂ -C ₆ -alkenyl, Ci-C ₆ -alkoxy, C ₂ -C ₆ -alkanoyl, Ci-C ₆ -alkylester, (mono- and di-Ci-C ₆ -alkylamino)C ₀ -C ₂ - alkyl, Ci-C ₂ -haloalkyl, and Ci-C ₂ haloalkoxy.

[0052] A "dosage form" means a unit of administration of an active agent. Examples of dosage forms include tablets, capsules, injections, suspensions, liquids, emulsions, creams, ointments, suppositories, inhalable forms, transdermal forms, and the like.

[0053] "Pharmaceutical compositions" are compositions comprising at least one active agent, such as a compound or salt, solvate, or hydrate of Formula (I) or a prodrug thereof, and at least one other substance, such as a carrier. Pharmaceutical compositions optional contain one or more additional active agents. When specified, pharmaceutical compositions meet the U.S. FDA's GMP (good manufacturing practice) standards for human or non-human drugs. "Pharmaceutical combinations" are combinations of at least two active agents which may be combined in a single dosage form or provided together in separate dosage forms with instructions that the active agents are to be used together to treat a disorder, such as a Gram-negative bacterial infection.

[0054] "Pharmaceutically acceptable salts" includes derivatives of the disclosed compounds in which the parent compound is modified by making inorganic and organic, non-toxic, acid or base addition salts thereof. The salts of the present compounds can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or the like), or by reacting free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are typically carried out in water or in an organic solvent, or in a mixture of the two. Generally, non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred, where practicable. Salts of the present compounds further include solvates of the compounds and of the compound salts.

[0055] Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts and the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, conventional non-toxic acid salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, H0 ₂ C-(CH ₂ ) _n -C0 ₂ H where n is 0-4, and the like. Lists of additional suitable salts may be found, e.g., in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985).

[0056] The term "carrier" applied to pharmaceutical compositions/combinations of the disclosure refers to a diluent, excipient, or vehicle with which an active compound is provided.

[0057] A "pharmaceutically acceptable excipient" means an excipient that is useful in preparing a pharmaceutical composition/ combination that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes an excipient that is acceptable for veterinary use as well as human pharmaceutical use. A "pharmaceutically acceptable excipient" as used in the present application includes both one and more than one such excipient. [0058] A "subject" is a human or non-human animal in need of medical treatment. Medical treatment can include treatment of an existing condition, such as a disease or disorder, prophylactic or preventative treatment, or diagnostic treatment. In some embodiments the subject is a human patient.

[0059] "Providing" means giving, administering, selling, distributing, transferring (for profit or not), manufacturing, compounding, or dispensing.

[0060] "Treatment," as used herein includes providing a compound of this disclosure such as a compound of any of Formulae (I), either as the only active agent or together with at least one additional active agent sufficient to: (a) inhibiting the disease, i.e. arresting its development; and (b) relieving the disease, i.e., causing regression of the disease and in the case of a bacterial infection to eliminate the infection in the subject. "Treating" and "treatment" also means providing a therapeutically effective amount of a compound of the disclosure as the only active agent or together with at least one additional active agent to a subject having or susceptible to a bacterial infection. "Prophylactic treatment" includes administering an amount of a compound of the disclosure sufficient to significantly reduce the likelihood of a disease from occurring in a subject who may be predisposed to the disease but who does not have it.

[0061] A "therapeutically effective amount" of a pharmaceutical composition/ combination is an amount effective, when administered to a subject, to provide a therapeutic benefit such as an amelioration of symptoms, e.g., an amount effective to decrease the symptoms of a bacterial infection and/ or effect a cure. In certain circumstances a subject suffering from a microbial infection may not present symptoms of being infected. Thus a therapeutically effective amount of a compound is also an amount sufficient to significantly reduce the detectable level of microorganism in the subject's blood, serum, other bodily fluids, or tissues. The disclosure also includes, in certain embodiments, using compounds of the disclosure in prophylactic treatment and therapeutic treatment. In the context of prophylactic or preventative treatment a "therapeutically effective amount" is an amount sufficient to significantly decrease the treated subject's risk of contracting a bacterial infection. For example, prophylactic treatment may be administered when a subject will knowingly be exposed to infectious microbes. A significant reduction is any detectable negative change that is statistically significant in a standard parametric test of statistical significance such as Student's T-test, where p < 0.05.

[0062] The term "prodrug" refers to compounds that are transformed in vivo to yield a disclosed compound or a pharmaceutically acceptable salt, hydrate or solvate of the compound. The transformation may occur by various mechanisms (such as by esterase, amidase, phosphatase, oxidative and or reductive metabolism) in various locations (such as in the intestinal lumen or upon transit of the intestine, blood or liver). Prodrugs are well known in the art (for example, see Nature Reviews Drug Discovery 2008, 7, 255; Current Topics in Medicinal Chemistry 2011, 11, 2265; and Molecules 2008, 13, 519). [0063] For example, if a compound of the disclosure or its pharmaceutically acceptable salt, hydrate or solvate contains a carboxylic acid functional group, a prodrug can comprise an ester formed by the replacement of the hydrogen atom of the acid group with a group such as Ci-C ₈ -alkyl, (C ₂ -Ci ₂ alkyl)carbonyloxymethyl, l-(alkylcarbonyloxy)ethyl having from 4 to 9 carbon atoms, 1 -methyl- 1- (alkylcarbonyloxy)-ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1 -(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-l- (alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms, l-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms, 3- phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl, di-N,N-(Ci-C ₂ -alkyl)amino(C ₂ -C ₃ -alkyl) (such as beta-dimethylaminoethyl), carbamoyl-(Ci-C ₂ -alkyl), N,N-di(Ci-C ₂ -alkyl)carbamoyl-(Ci-C ₂ -alkyl), and piperidino-, pyrrolidino- or morpholino-(C ₂ -C ₃ -aikyl).

[0064] Similarly, if a compound of the disclosure contains an alcohol functional group, a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a group such as (Ci-C ₆ -alkyl)carbonyloxymethyl, l-((Ci-C ₆ -alkyl)carbonyloxy)ethyl, l-methyl-l-((Ci-C ₆ - alkyl)carbonyloxy)ethyl, (Ci-C6-alkoxy)carbonyloxymethyl, N-(Ci-C6-alkoxy)carbonylaminomethyl, succinoyl, (d-Cs-alkyftcarbonyl, -P(0)(OH) ₂ , -P(0)(OH)(0-C _! -C ₆ -alkyl), _, -P(0)(0-Ci-C ₆ -alkyl) _2> glycosyl (the radical resulting from the removal of a hydroxyl group of the hemiacetal form of a carbohydrate) or alpha-aminoacyl, wherein the alpha-aminoacyl is derived from the natural L-aminoacids.

[0065] If a compound of the disclosure incorporates an amine functional group, a prodrug can be formed, for example, by creation of an amide or carbamate, an as (C ₁ -C6- alkyl)carbonyloxymethylcarbonyl derivative, a l-((Ci-C6-alkyl)carbonyloxyethylcarbonyl) derivative, an (oxodioxolenyl)methyl derivative, a N-Mannich base, imine or enamine. In addition, a secondary amine can be metabolically cleaved to generate a bioactive primary amine, or a tertiary amine can metabolically cleaved to generate a bioactive primary or secondary amine. For examples, see Simplicio, et al,

Molecules 2008, 13, 519 and references therein.

CHEMICAL DESCRIPTION

[0066] Formulae (I) include all subformulae thereof. In certain situations, the compounds of any of Formulae (I) may contain one or more asymmetric elements such as stereogenic centers, stereogenic axes and the like, e.g. asymmetric carbon atoms, so that the compounds can exist in different stereoisomeric forms. These compounds can be, for example, racemates or optically active forms. For compounds with two or more asymmetric elements, these compounds can additionally be mixtures of diastereomers. For compounds having asymmetric centers, it should be understood that all of the optical isomers and mixtures thereof are encompassed. In addition, compounds with carbon-carbon double bonds may occur in Z- and E-forms, with all isomeric forms of the compounds being included in the present disclosure. In these situations, single enantiomers, i.e., optically active forms, can be obtained by asymmetric synthesis, synthesis from optically pure precursors, or by resolution of the racemates.

Resolution of the racemates can also be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example using a chiral HPLC column.

[0067] Where a compound exists in various tautomeric forms, the disclosure is not limited to any one of the specific tautomers, but rather includes all tautomeric forms.

[0068] Certain compounds are described herein using a general formula that includes variables, e.g. Ri- R ₉ . Unless otherwise specified, each variable within such a formula is defined independently of other variables. Thus, if a group is said to be substituted, e.g. with 0-2 R*, then the group may be substituted with up to two R* groups and R* at each occurrence is selected independently from the definition of R*. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

[0069] In addition to compounds and salts of Formula (I) disclosed in the SUMMARY section, the disclosure includes embodiments in which any of the following conditions are met

[0070] (Tt Y^s CH;

Y ² and Y ⁴ are independently selected from CH and C(R ² );

Y ³ is selected from N, CH and C(R ² );

Y ⁵ is selected from N and C(R ² ); and

Z is N(H).

[0071] (ii) Y ² is selected from CH, C(halo) and C(CN);

Y ³ is selected from N, CH and C(halo);

Y ⁴ is selected from CH, C(d-C ₄ alkyl) and C(C ₃ -C ₆ cycloalkyl); and

Y ⁵ is selected from N, C(CN), C(C _! -C ₄ alkyl) and C(C ₃ -C ₆ cycloalkyl), wherein any alkyl or cycloalkyl in Y ⁴ or Y ⁵ is optionally and independently substituted with halo or hydroxyl.

[0072] (iii) Y ² is selected from CH, C(C1), C(F), and C(CN);

Y ³ is selected from N, CH and C(C1);

Y ⁴ is selected from CH, C(CH(CH ₃ ) ₂ OH) and C(l-hydroxycycloprop-l-yl); and

Y ⁵ is selected from N, C(CN), C(CH(CH ₃ ) ₂ OH) and C(l-hydroxycycloprop-l-yl).

[0073] (iv) Y ² is C(C1).

[0074] (v) X is selected from -S-, -NH- and -CH ₂ .

[0075] (vi) Each R ^la is independently selected from hydrogen and hydroxyl; and each R ^lb is hydrogen. [0076] (vii) Each of Y ⁶ , Y ⁷ , Y ⁸ , and and Y ⁹ is independently selected from CH and C(R ² ).

(viii) Y ⁸ is C(R ² ).

[0077] (ix) Y ⁸ is a carbon atom substituted with a substituent selected from halo, -R ⁶ -heteroaryl, and -R ⁶ -aryl, wherein R ⁶ is selected from -0-, -CH ₂ - and -CF ₂ -; and the heteroaryl or aryl moiety in Y ⁸ is optionally substituted with one or more substituents independently selected from halo, cyano and Ci-C ₄ alkyl. In certain embodiments the heteroaryl or aryl is a phenyl, pyridine, pyrimidine, or pyrazine, each of which may be optionally substituted, for example with one or more halo, cyano and C ₁ -C4 alkyl.

[0078] (x) Y ⁸ is a carbon atom substituted with a substituent selected from chloro, pyridin-3- yloxy, pyridin-3-ylmethyl, difluoro(pyridin-3-yl)methyl, pyrimidin-2-yloxy, pyrazin-2-yloxy, and phenoxy, wherein any ring moiety in a substituent on Y ⁸ is optionally substituted with one or two substituents independently selected from fluoro, chloro, cyano, and methyl.

[0079] (xi) Each R ³ , if present, is independently selected from hydrogen, methyl, ethyl and -CH ₂ -C(CH ₃ ) ₂ OH.

[0080] (xii) R ⁴ , if present, is hydrogen.

[0081] (xiii) Q is selected from 6-chloro-lH-indazol-3-yl, 6-chloro-l-methylindazol-3-yl, 6- chloro-lH-indol-3-yl, 6-chloro-l-methylindol-3-yl, 6-(pyridin-3-yloxy)-lH-pyrrolo[2,3-b]pyridin-3-yl, 6- (pyridin-3-ylmethyl)-lH-pyrrolo[2,3-b]pyridin-3-yl, 6-(pyridin-3-yloxy)imidazo[l,5-a]pyridin-l-yl, 7- (pyridin-3-yloxy)imidazo[l,2-a]pyridin-3-yl), 3-(pyridin-3-yloxy)imidazo[l,5-a]pyrimidin-8-yl, 6- (difluoro(pyridin-3 -yl)methyl)- 1 H-indol-3-yl, 6-(pyridin-3 -yloxy)- 1 H-indol-3 -yl, 6-(pyridin-3 -yloxy)- 1 H- indazol-3-yl, 6-(pyridin-3-yloxy)-l -methylindol-3-yl, 6-(pyridin-3-yloxy)-l -methylindazol-3-yl, 6- (pyriazin-2-yloxy)-lH-indol-3-yl, 6-(2-chloropyridin-3-yloxy)-lH-indol-3-yl, 6-(2-methylpyridin-3- yloxy)-lH-indol-3-yl, 6-(2-chloro-6-methylpyridin-3-yloxy)-lH-indol-3-yl, 6-(2,6-dimethylpyridin-3- yloxy)-lH-indol-3-yl, 6-(pyridin-3-yloxy)-5-fluoro-lH-indol-3-yl, 6-(pyridin-3-yloxy)-l-(2-hydroxy-2- methylpropyl)-indazol-3-yl, 6-(pyridin-3-yloxy)-l-ethylindol-3-yl, 6-(5-fluoropyridin-3-yloxy)-lH-indol- 3-yl, 6-(5-fluoro-6-chloropyridin-3-yloxy)-lH-indol-3-yl, 6-(2-cyanophenoxy)-lH-indol-3-yl, 6-(3- cyanophenoxy)-lH-indol-3-yl, 6-(4-cyanophenoxy)-lH-indol-3-yl, 6-(pyridin-3-yloxy)-l-(2-hydroxy-2- methylpropyl)-indol-3-yl, 6-(pryimidin-2-yloxy)-lH-indol-3-yl, and 6-(pyridin-3-yloxy)-l-ethylindazol- 3-yl.

[0082] (xiv) W is C and, when X is a bond or -C(R ^la )(R ^lb )- and Y ⁵ is CH or C(R ² ), W is additionally selected from N.

[0083] (xv) X is NH and R ^la and R ^lb are both hydrogen.

[0084] (xvi) X is CH ₂ and R ^la and R ^lb are both hydrogen.

[0085] (xvii) X is S and R ^la and R ^lb are both hydrogen. [0086] The disclosure includes any and all combinations of conditions (i) to (xvii) that result in a stable compound of Formula (I). In any of the conditions (i) to (xvii) where a variable of Formula (I) is selected from a list of moieties, the disclosure also includes individual and subsets of moieties from that list in both that condition and in combination with any and all combinations of the other of conditions (i) to (xiii). Moreover, the disclosure also includes combination comprising individual or subsets of moieties from one of conditions (i) to (xvii) combined with individual or subsets of moieties from one or more other of conditions (i) to (xvii) and optionally in combination with additional conditions (i) to (xvii). As a non-limiting example of the foregoing, the disclosure includes a compound wherein X is -S- (an individual moiety in condition (v)); Y ⁸ is a carbon atom substituted with a substituent selected from pyridin-3-yloxy, pyridin-3-ylmethyl and pyrimidin-2-yloxy, wherein any ring moiety in a substituent on Y ⁸ is optionally substituted with one or two substituents independently selected from fluoro, chloro, cyano, and methyl (a subset of the moieties set forth in condition (x)); and each R ³ , if present, is independently selected from hydrogen, methyl, ethyl and -CH ₂ -C(CH ₃ ) ₂ OH (the entirety of condition (xi)).

[0087] Formula I includes the following subformulas in which Q, R ^la , R ^lb , and R ² carry any of the defi

[0088] The disclosure includes prodrugs of Formula I. For example the disclosure includes prodrugs of the following formulas:

where each of the variables, e.g. Y^Y ¹¹ , R ^la , R ^lb , and R ⁵ may carry any of the definitions set forth in this disclosure.

[0089] L ¹ is a bond or a Ci-C ₆ -alkyl, C ₂ -C ₆ alkenyl, or C ₂ -C ₆ alkynyl linker, and one or more methylene units in the alkyl, alkenyl, or alkynyl portion are optionally and independently replaced with O , S, N(R ⁶ ) , S(O) or S(0) ₂ ;

L ² is a Ci-C6-alkyl, C ₂ -C ₆ alkenyl, or C ₂ -Cealkynyl linker, and one or more methylene units in the alkyl, alkenyl, or alkynyl portion are optionally and independently replaced with O , S, N(R ⁶ ) , S(O) or S(0) ₂ ; and

PD is a water-solublizing prodrug well known in the art such as but not limited to P(0)(OH) ₂ or an alpha-aminoacyl, wherein the alpha-aminoacyl is derived from the natural L-aminoacids.

[0090] The disclosure includes, but is not restricted to, the following prodrugs:



METHODS OF TREATMENT

[0091] The disclosure includes a method of treating a bacterial infection in a subject by administering an effective amount of one or more compounds of the disclosure to a subject at risk for a bacterial infection or suffering from a microorganism infection. Treatment of human patients is particularly contemplated. However, treatment of non-human subjects is within the scope of the disclosure. The disclosure includes treatment or prevention of microbial infections in fish, amphibians, reptiles or birds, but a preferred embodiment of the disclosure includes treating mammals.

[0092] In some embodiments, the bacterial infection or antibiotic-tolerant infection is caused by a Gram-negative bacterium.

[0093] In an embodiment of any of the methods of this disclosure, the microbial infection is the result of a pathogenic bacterial infection. Examples of pathogenic bacteria include, without limitation, bacteria within the genera Aerobacter, Aeromonas, Acinetobacter, Agrobacterium, Bacillus, Bacteroides, Bartonella, Bordetella, Brucella, Burkholderia Calymmatobacterium, Campylobacter, Citrobacter, Clostridium, Corynebacterium, Enterobacter, Enterococcus, Escherichia, Francisella, Haemophilus, Hafnia, Helicobacter, Klebsiella, Legionella, Listeria, Morganella, Moraxella, Proteus, Providencia, Pseudomonas, Salmonella, Serratia, Shigella, Staphylococcus, Streptococcus, Treponema, Xanthomonas, Vibrio, and Yersinia. Specific examples of such bacteria include Vibrio harveyi, Vibrio cholerae, Vibrio parahemolyticus, Vibrio alginolyticus, Pseudomonas phosphoreum, Pseudomonas aeruginosa, Yersinia enterocolitica, Escherichia coli, Salmonella typhimurium, Haemophilus influenzae, Helicobacter pylori, Bacillus subtilis, Borrelia burgdorferi, Neisseria meningitidis, Neisseria gonorrhoeae, Yersinia pestis, Campylobacter jejuni, Mycobacterium tuberculosis, Enterococcus faecalis, Streptococcus pneumoniae, Streptococcus pyogenes, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus epidermidis, and Staphylococcus aureus.

[0094] In some embodiments, the gram-negative bacterium is a Pseudomonas, e.g., P.

aeruginosa.

[0095] In some embodiments, the gram-negative bacterium is Burkholderia species.

[0096] In some embodiments, the gram-negative bacterium is Acinetobacter, e.g., A. baumannii.

[0097] In some embodiments, the gram-negative bacterium is an Enterobacteriaceae, e.g., Klebsiella pneumonia, e.g., Escherichia coli, e.g., Enterobacter cloacae, e.g., Serratia marcescens, e.g., Salmonella typhimurium, e.g., Shigella dysenteriae, e.g., Proteus mirabilis, e.g., Citrobacter freundii, e.g., Yersinia pestis.

[0098] In some embodiments, the infection is a polymicrobial infection, e.g., an infection comprising more than one organism. In some embodiments, the infection comprises at least one of the organisms listed above, e.g., one or more oi Pseudomonas, e.g., P aeruginosa, Kelbsiella, e.g., Klebsiella pneumoniae, and/or Acinetobacter, e.g., A. baumannii.

[0099] In some embodiments, the methods further include administering an additional active agent in combination with a compound of the disclosure, such as an antibiotic selected from the group consisting of: beta-lactams such as penicillins, cephalosporins, carbacephems, cephamycins,

carbapenems, monobactams, quinolones including fluoroquinolones and similar DNA synthesis inhibitors, tetracyclines, aminoglycosides, macrolides, glycopeptides, chloramphenicols, glycylcyclines, lincosamides, lipopeptides, and oxazolidinones.

[00100] In some embodiments, the bacterial infection is an upper respiratory tract infection, pneumonia, a systemic infection, sepsis and septic shock, a urinary tract infection, a gastrointestinal infection, endocarditis, a bone infection, central nervous system infections such as meningitis, or an infection of the skin and soft tissue.

[00101] In some embodiments, the subject is a mammal, e.g., a human or non-human mammal. In some embodiments, the methods include treating one or more cells, e.g., cells in a culture dish. [00102] In one aspect, the present disclosure features a method of treating a Gram-negative infection in a subject, the method comprising administering to said subject in need of such treatment a therapeutically effective amount of a compound described herein.

[00103] In some embodiments, the Gram-negative infection is caused by Pseudomonas aeruginosa.

[00104] In other embodiments the disclosure includes treating an infection caused by Gram positive bacteria, such as Staphylococcus epidermidis and Staphylococcus aureus.

[00105] In some embodiments, the subject is a trauma patient or a burn patient suffering from a burn or skin wound.

[00106] In a further aspect, the present disclosure features a method of reducing bacterial tolerance in a subject, the method comprising administering to said subject a therapeutically effective amount of a compound described herein.

[00107] In some embodiments, the method further includes identifying said subject suffering from an infection with bacteria tolerant to antimicrobial therapy.

[00108] The disclosure includes methods of treatment in which a compound or composition of the disclosure is administered orally, topically, parenterally, or inhaled.

[00109] A compound of the disclosure may be administered about 1 to about 5 times per day. Daily administration or post-periodic dosing may be employed. Frequency of dosage may also vary depending on the compound used, the particular disease treated and the bacteria causing the disease. It will be understood, however, that the specific dose level for any particular subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease undergoing therapy.

AGRICULTURAL METHODS

[00110] The disclosure also includes methods of treating bacterial infection in plants and fungal crops (e.g. mushrooms) comprising contacting a compound of the Formula I with a plant or fungal organism.

[00111] The plant may be an agricultural crop plant, such as a tobacco plant or a tomato plant .For example Pseudomonas species also can be involved in agricultural damage. Some of Pseudomonas syringae's numerous pathovars can be plant pathogens. For example P. syringae pathovar tabacii, phaseolicola, and tomato are pathogenic to plants. Pseudomonas agarici and Pseudomonas

tolaasii are pathogens of cultivated mushrooms. The compounds of this disclosure can are useful for reducing or eliminating the virulence of such Pseudomonal pathovars to plant and fungal crops. PHARMACEUTICAL COMPOSITIONS

[00112] The disclosure includes the process for making a pharmaceutical composition containing at least one compound of Formula I. An embodiment comprises mixing one or more of the present compounds and an optional pharmaceutically acceptable carrier; and includes those compositions resulting from such a process, which process includes conventional pharmaceutical techniques.

[00113] The compositions of the disclosure include ocular, oral, nasal, transdermal, topical with or without occlusion, intravenous (both bolus and infusion), inhalable, and injection (intraperitoneally, subcutaneously, intramuscularly, intratumorally, or parenterally) formulations. The composition may be in a dosage unit such as a tablet, pill, capsule, powder, granule, liposome, ion exchange resin, sterile ocular solution, or ocular delivery device (such as a contact lens and the like facilitating immediate release, timed release, or sustained release), parenteral solution or suspension, metered aerosol or liquid spray, drop, ampoule, auto-injector device, or suppository; for administration ocularly, orally, intranasally, sublingually, parenterally, or rectally, or by inhalation or insufflation.

[00114] The dosage form containing the composition of the disclosure contains an effective amount of the active ingredient necessary to provide a therapeutic effect by the chosen route of administration. The composition may contain from about 5,000 mg to about 0.5 mg (preferably, from about 1 ,000 mg to about 0,5 mg) of a compound of the disclosure or salt form thereof and may be constituted into any form suitable for the selected mode of administration.

[00115] In one embodiment of the disclosure, the pharmaceutical composition may be a parenteral formulation suitable for parenteral administration via injection or infusion. A parenteral formulation may consist of the active ingredient dissolved in or mixed with an appropriate inert liquid carrier. Acceptable liquid carriers usually comprise aqueous solvents and other optional ingredients for aiding solubility or preservation. Such aqueous solvents include sterile water, Ringer's solution, or an isotonic aqueous saline solution. Other optional ingredients include vegetable oils (such as peanut oil, cottonseed oil, and sesame oil), and organic solvents (such as solketal, glycerol, and formyl). A sterile, non-volatile oil may be employed as a solvent or suspending agent. The parenteral formulation is prepared by dissolving or suspending the active ingredient in the liquid carrier whereby the final dosage unit contains from 0.005 to 10% by weight of the active ingredient. Other additives include preservatives, isotonizers, solubilizers, stabilizers, and smoothing agents. Injectable suspensions may also be prepared, in which case appropriate liquid carriers, suspending agents and the like may be employed.

[00116] In another embodiment, the pharmaceutical composition of this disclosure may be a composition formulated for administration directly to the lungs by inhalation, such as an aerosol formulation. Known aerosol drug delivery systems include, for example, a unit dose dry-powder inhaler, a dry powder pulmonary device, a pressurized metered dose inhaler, a metered-dose inhaler, a nebulizer, and the like.

[00117] In another embodiment, the pharmaceutical compositions of this disclosure may be formulated for oral administration. Compositions of the disclosure suitable for oral administration include solid forms such as pills, tablets, caplets, capsules (each including immediate release, timed release, and sustained release formulations), granules and powders; and, liquid forms such as solutions, syrups, elixirs, emulsions, and suspensions.

[00118] In another embodiment, the pharmaceutical compositions of this disclosure may formulated for topical administration, including topical administration to the eye. For ocular

administration, the composition is preferably in the form of an ophthalmic composition. The ophthalmic compositions are preferably formulated as eye -drop formulations and filled in appropriate containers to facilitate administration to the eye, for example a dropper fitted with a suitable pipette.

EXAMPLES ABBREVIATIONS

TEA Triethylamine TLC Thin layer chromatography

TFA trifluoroacetic acid UPLC Ultra performance liquid

chromatography

TMSOTf Trimethylsilyl Triflate

THF Tetrahydrofuran

BIOLOGICAL METHODS

[00119] Experiments were performed to identify compounds that inhibit the MvfR regulon without altering growth, ultimately attenuating infection. MvfR is a LysR-type transcriptional regulator that directs 4-hydroxy-2-alkylquinolines (HAQs) synthesis, including that of its ligands,

4-hydroxy-2-heptylquinoline (HHQ) and 3,4-dihydroxy-2-heptylquinoline (PQS). MvfR regulates the production of many virulence factors including pyocyanin, one of the many toxins secreted by

Pseudomonas aeruginosa. Both MvfR and PQS have been demonstrated as essential for pathogenesis in several host models.

[00120] MvfR promotes the production of HAQs by binding to and activating the pqs operon, which encodes enzymes for HAQ synthesis. Anthranilic acid (AA), derived from the phnAB, kynABU, and trpEG pathways, is the precursor for HAQs. Pqs A encodes an anthranilate -coenzyme A ligase, which activates anthranilic acid and catalyzes the first committed step to HAQ production. The exact roles of PqsB and PqsC are unknown, though both show homology to acyl-carrier-proteins and both are required for HHQ and PQS production. PqsD is a condensing enzyme that along with PqsA has been shown to be necessary and sufficient for the production of 2,4-dihydroxyquinoline (DHQ), a molecule whose biological role has yet to be determined. The final gene of the operon, PqsE encodes for a putative hydrolase, and while the protein is not required for the synthesis of HAQs, it is necessary for pyocyanin production.

[00121] Inhibition of Pyocyanin production is correlated with reduced P. aeruginosa infectivity. HHQ and PQS inhibition is also correlated with reduced P. aeruginosa infectivity. PQS inhibition is correlated with reduced infectivity of other bacterial pathogens as PQS is known to affect oxygen consumption and cell to cell communication of other Gram-negative and Gram-positive bacteria (Toyofuku, M. et al., Microbes Environ. (2010) 25(1): 1-7). EXAMPLE 1. SYNTHESIS OF 2-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YLTHIO)-1-(6-CHLORO-1H-IND AZOL- 3 - YL)ETH ANONE (Compound 1)

[00122] Step A. 6-Chloro-lH-indazole. To a mixture of 4-chloro-2-fTuorobenzaldehyde (7.9 g, 50 mmol) in DMA (50 mL) at 10 °C was added dropwise N ₂ H ₄ .H ₂ 0 (25 mL, 99% wt). After addition, the mixture was placed in a sealed tube and stirred at r.t. for 1 hr, then heated to 100 °C and stirred at that temperature for 48 hr. The resulting mixture was cooled to r.t., poured into ice water, and extracted with EtOAc. The combined organic layers were washed twice with brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-chloro-lH-indazole (3.2 g, 42% yield) as pale yellow solid. LC-MS: m/z 153 (M+H) ⁺ .

[00123] Step B. 6-Chloro-3-iodo-lH-indazole. To a mixture of 6-chloro-lH-indazole (3.0 g, 20 mmol) in DMF (50 mL) at r.t. was added I ₂ (10 g, 40 mmol), followed by addition of well-powdered KOH (4.5 g, 80 mmol). The reaction mixture was stirred at r.t. for 3 hr, then poured into a pre-cooled solution of aq.Na ₂ S ₂ 0 ₃ and extracted with EtOAc. The combined organic layers were washed with aq. LiCl (10%wt), dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-chloro-3-iodo-lH-indazole (4.9 g, 88.1% yield) as white solid. LC-MS: m/z 279 (M+H) ⁺ .

[00124] Step C. Methyl 6-chloro-lH-indazole-3-carboxylate. A mixture of 6-chloro-3-iodo-lH- indazole (2.8 g, 10 mmol), Pd (dppf)Cl ₂ (245 mg, 0.3 mmol), dppf (220 mg, 0.4 mmol), and Et ₃ N (1.7 g, 17 mmol) in a mixed solvent of toluene (30 mL) and MeOH (30 mL) was stirred at 70 °C under an atmosphere of CO (3 bar pressure) for 12 hr. The resulting mixture was cooled to r.t. and concentrated under reduced pressure. The residue was purified by flash column chromatography to give methyl 6- chloro-lH-indazole-3-carboxylate (1.7 g, 80.9% yield) as gray solid. LC-MS: m/z 211 (M+H) ⁺ . [00125] Step D. 6-Chloro-lH-indazole-3-carboxylic acid. To a mixture of methyl 6-chloro-lH- indazole-3-carboxylate (1.7 g, 8.1 mmol) in MeOH (90 mL) at r.t. were added THF (30 mL) and a solution of NaOH (2.4 g, 60 mmol) in H ₂ 0 (30 mL). The resulting mixture was stirred at reflux for 8 hr, then cooled to r.t. and concentrated to a volume of about 30 mL under reduced pressure. The residue was diluted with water (30 mL) and washed with DCM. The aq. phase was separated and acidified to pH 5-6 with cone. HCl at 0 °C. The resulting precipitates were collected by filtration, washed with H ₂ 0 and dried under high vacuum to give 6-chloro-lH-indazole-3-carboxylic acid (1.2 g, 75.9% yield) as gray solid. LC-MS: m/z 197 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 13.92 (s, 1H), 13.21 (br, 1 H), 8.07 (d, = 8.4 Hz, 1H), 7.75 (s, 1H), 7.31 (d, = 8.4 Hz, 1H).

[00126] Step E. 6-Chloro-N-methoxy-N-methyl-lH-indazole-3-carboxamide.To a mixture of 6- chloro-lH-indazole-3-carboxylic acid (1 g, 5 mmol) in THF (50 mL) at r.t. were added Ν,Ο- dimethylhydroxylamine hydrochloride (0.58 g, 6 mmol), pyridine (2.1 mL, 26 mmol) and EDCI (1.73 g, 9 mmol). The resulting mixture was stirred at r.t. for 20 hr, then poured into water and extracted with EtOAc. The combined organic layers were washed in sequence with water, aq. HCl (0.5 N) and brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give crude 6-chloro-N- methoxy-N-methyl-lH-indazole-3-carboxamide (1.2 g, quant.crude yield) as gray solid which was used directly in the next step without any further purification. LC-MS: m/z 240 (M+H) ⁺ .

[00127] Step F. l-(6-chloro-lH-indazol-3-yl)ethanone. To a mixture of the above crude 6- chloro-N-methoxy-N-methyl-lH-indazole-3-carboxamide (1.2 g, 5 mmol) in dry THF (60 mL) at -5 °C was added dropwise MeMgBr (7 mL, 21 mmol, 3 M in THF) over a period of 10 min. After addition was complete, the mixture was stirred at 0°C for 2 hr, thenslowly poured into a pre-cooled solution of NH ₄ C1 with vigorous stirring and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column

chromatography to give l-(6-chloro-lH-indazol-3-yl)ethanone (600 mg, 60% yield) as white solid.

LC-MS: m/z 195(M+H) ⁺ .

[00128] Step G. 2-bromo-l-(6-chloro-lH-indazol-3-yl)ethanone. To a mixture of l-(6-chloro- lH-indazol-3-yl)ethanone (100 mg, 0.5 mmol) in EtOAc (5 mL) was added CuBr ₂ (279 mg, 1.25 mmol). The reaction mixture was stirred at reflux for 2 hr, then cooled to r.t. and filtered through Celite. The filtrate was washed with a solution of aq. Na ₂ S ₂ 0 ₃ and brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give crude 2-bromo-l-(6-chloro-lH-indazol-3-yl)ethanone (150 mg, quant, crude yield) as pale yellow solid which was used directly in the next step without any further purification. LC-MS: m/z 273(M+H) ⁺ .

[00129] Step H. 2-(5-Chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6-chloro-lH-ind azol-3- yljethanone. To a mixture of the above crude 2-bromo-l-(6-chloro-lH-indazol-3-yl)ethanone (70 mg, 0.25 mmol) in DMF (3 mL) were added 5-chloro-lH-benzo[d]imidazole-2-thiol (70 mg, 0.375 mmol) and K ₂ C0 ₃ (138 mg, 1 mmol). The reaction mixture was stirred at r.t. for 2 hr, then poured into ice water and extracted with DCM. The combined organic layers were washed with aq. LiCl (10 wt), dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 2-(5-chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6-chloro-lH-ind azol-3-yl)ethanone (30 mg, 31.8% yield) as white solid. LC-MS: m/z377 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 13.5 (br, 1H), 8.14 (d, J = 8.8 Hz, 1H), 7.85 (s, 1H), 7.47 (s, 1H), 7.36 - 7.42 (m, 2H), 7.11-7.13 (m, 1H), 5.11 (s, 2H).

EXAMPLE 2. SYNTHESIS OF 2-(2-(6-CHLORO-1H-INDOL-3-YL)-2-OXOETHYLTHIO)-1H- -5-CARBONITRILE (Compound 2)

[00130] Step A. 2-Mercapto-lH-benzo[d]imidazole-5-carbonitrile. To a mixture of 3,4- diaminobenzonitrile (1.6 g, 12 mmol) in EtOH (49 mL) and water (7 mL) was added potassium O-ethyl carbonodithioate (2.3 g, 14.4 mmol). The reaction mixture was stirred at reflux for 4 hr, followed by addition of charcoal (200 mg). The resulting mixture was stirred at reflux for another 10 min, then cooled and filtered. The filtrate was concentrated under reduced pressure. The residue was triturated with water (100 mL) and filtered. The solid was collected and dried under high vacuum to afford 2-mercapto-lH- benzo[d]imidazole-5-carbonitrile (1.2 g, 57% yield) as brown solid.LC-MS: m/zl76(M+H) ⁺ .

[00131] S¾77 B. 2-Chloro-l-(6-chloro-lH-indol-3-yl)ethanone. To a mixture of 6-chloro-lH- indole (304 mg, 2 mmol) in DCM (10 mL) at 0 °C was added A1C1 ₃ (266 mg, 2 mmol), followed by slow addition of 2-chloroacetyl chloride (226 mg, 2 mmol). The reaction mixture was stirred at 0 °C for 15 min, then quenched with aq. HC1 (10%wt, 4 mL). The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ , and concentrated to under reduced pressure to afford 2-chloro-l-(6-chloro-lH-indol-3- yl)ethanone (410 mg) with 50% purity as a yellow solid. LC-MS: m/z 228(M+H) ⁺ .

[00132] Step C. 2-(2-(6-chloro-lH-indol-3-yl)-2-oxoethylthio)-lH-benzo[d]imi dazole-5- carbonitrile. To a mixture of 2-mercapto-lH-benzo[d]imidazole-5-carbonitrile (88 mg, 0.5 mmol) in DMF (8 mL) were added K ₂ C0 ₃ (138 mg, 1 mmol) and 2-chloro-l-(6-chloro-lH-indol-3-yl)ethanone (114 mg, 0.5 mmol). The reaction mixture was stirred at r.t. for 1 hr, then quenched with aq. LiCl (10 wt) and extracted with DCM. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford 2-(2-(6-chloro- lH-indol-3-yl)-2-oxoethylthio)-lH-benzo[d]imidazole-5-carbon itrile (18 mg, 11% yield) as white solid. LC-MS: m/z 367(M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.33 (s, 1H), 8.62 (s, 1H), 8.11 (d, = 8.5 Hz, 1H), 7.92 (s, 1H), 7.55 (d, = 8.1 Hz, 2H), 7.48 (d, = 8.2 Hz, 1H), 7.23 (d, = 8.5 Hz, 1H), 4.92 (s, 2H).

[00133] The procedure set forth above was used to produce the following compounds. 3 and 4 using the appropriate starting materials.

[00134] 2-((5 -chloro- 1 H-benzo[d]imidazol-2-yl)thio)- 1 -(6-chloro- 1 H-indol-3-yl)ethanone

[00135] LC-MS: m/z 376(M+H) H NMR (400 MHz, DMSO-d ₆ ) δ 12.51 (s, 1H), 8.61 (s, 1H), 8.11 (d, = 8.5 Hz, 1H), 7.57 (s, 1H), 7.47 (s, 1H), 7.42 (d, = 8.5 Hz, 1H), 7.23 (d, = 8.6 Hz, 1H), 7.12 (d, = 8.3 Hz, 1H), 4.88 (s, 2H).

[00136] 1 -(6-Chloro-l -methyl- lH-indol-3-yl)-2-(5-chloro- lH-benzo[d]imidazol-2-

[00137] LC-MS: m/z 390 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.84 (s, 1H), 8.63 (s, 1H), 8.11 (d, = 8.5 Hz, 1H), 7.76 (d, = 1.7 Hz, 1H), 7.52 - 7.37 (m, 2H), 7.28 (dd, = 8.5, 1.8 Hz, 1H), 7.13 (d, = 8.5 Hz, 1H), 4.85 (s, 2H), 3.90 (s, 3H).

EXAMPLE 3. SYNTHESIS OF 1-(6-CHLORO-1-METHYL-1H-INDAZOL-3-YL)-2-(5-CHLORO-1H- BENZO [D] IMID AZOL-2- YLTHIO)ETH ANONE (Compound 5)

[00138] Step A. 6-Chloro-lH-indazole. To a solution of 4-chloro-2-fluorobenzaldehyde (7.9 g, 50 mmol) in DMA (50 mL) in a sealed tube at 10 °C was added dropwise N ₂ H ₄ .H ₂ 0 (25 mL, 99 wt). The reaction mixture was stirred at r.t. for 1 hr then at 100 °C for 48 hr. The resulting mixture was cooled to r.t., poured into ice water and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-chloro-lH-indazole (3.2 g, 42% yield) as pale yellow solid. LC-MS: m/z 153 (M+H) ⁺ .

[00139] Step B. 6-Chloro-3-iodo-lH-indazole. To a mixture of 6-chloro-lH-indazole (3.0 g, 20 mmol) in DMF (50 mL) at r.t. was added I ₂ (10 g, 40 mmol), followed by addition of powdered KOH (4.5 g, 80 mmol). The reaction mixture was stirred at r.t for 3 hr, poured into a cooled solution of aq. Na ₂ S ₂ 0 ₃ and extracted with EtOAc. The combined organic layers were washed with aq. LiCl (10%wt), and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-chloro-3-iodo-lH-indazole (4.9 g, 88.1% yield) as white solid. LC-MS: m/z 279 (M+H) ⁺ .

[00140] Step C. Methyl 6-chloro-lH-indazole-3-carboxylate.A mixture of 6-chloro-3-iodo-lH- indazole (2.8 g, 10 mmol), Pd(dppf)Cl ₂ (245 mg, 0.3 mmol), dppf (220 mg, 0.4 mmol), and Et ₃ N (1.7 g, 17 mmol) in a mixed solvent of toluene (30 mL) and MeOH (30 mL) was stirred at 70 °C under an atmosphere of CO (3 bar pressure) for 12 hr. After cooling to r.t, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography to give methyl 6- chloro-lH-indazole-3-carboxylate (1.7 g, 80.9% yield) as gray solid. LC-MS: m/z 211 (M+H) ⁺ . [00141] Step D. 6-Chloro-lH-indazole-3-carboxylicacid. To a solution of methyl 6-chloro-lH- indazole-3-carboxylate (1.7 g, 8.1 mmol) in MeOH (90 mL) at r.t. were added THF (30 mL) and a solution of NaOH (2.4 g, 60 mmol) in H ₂ 0 (30 mL). The reaction mixture was stirred at reflux for 8 hr. After cooling to r.t, the mixture was concentrated under reduced pressure to a volume of about 30 mL. The resulting mixture was diluted with water (30 mL), washed with DCM, and then acidified to pH 5-6 with cone. HC1 at 0 °C. The solid was collected by filtration, washed with H ₂ 0 and dried under high vacuum to give 6-chloro-lH-indazole-3-carboxylic acid (1.2 g, 75.9% yield) as gray solid. LC-MS: m/z 197 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 13.92 (s, 1H), 13.21 (br, 1 H), 8.07 (d, = 8.4 Hz, 1H), 7.75 (s, 1H), 7.31 (d, = 8.4 Hz, 1H).

[00142] Step E. 6-Chloro-N-methoxy-N-methyl-lH-indazole-3-carboxamide. To a mixture of 6- chloro-lH-indazole-3-carboxylic acid (1 g, 5 mmol) in THF (50 mL) at r.t. were added Ν,Ο- dimethylhydroxylamine hydrochloride (0.58 g, 6 mmol), pyridine (2.1 mL, 26 mmol) and EDCI (1.73 g, 9 mmol). The reaction mixture was stirred at r.t. for 20 hr, then poured into water and extracted with EtOAc. The combined organic layers were washed in sequence with water, aq. HC1 (0.5 N) and brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give crude 6-chloro-N- methoxy-N-methyl-lH-indazole-3-carboxamide (1.2 g, quant, crude yield) as gray solid which was used directly in the next step. LC-MS: m/z 240 (M+H) ⁺ .

[00143] Step F. l-(6-Chloro-lH-indazol-3-yl)ethanone. To a solution of the above crude 6- chloro-N-methoxy-N-methyl-lH-indazole-3-carboxamide (1.2 g, 5 mmol) in dry THF (60 mL) at -5 °C was added dropwise MeMgBr (7 mL, 21 mmol, 3 M in THF) over 10 min. After addition, the mixture was stirred at 0°C for 2 hr, then slowly poured into an ice -cooled solution of aq. NH ₄ C1 with vigorous stirring. The resulting mixture was extracted with EtOAc, and the combined organic layers were concentrated under reduced pressure. The residue was purified by flash column chromatography to give l-(6-chloro-lH-indazol-3-yl)ethanone (600 mg, 60% yield) as white solid. LC-MS: m/z 195(M+H) ⁺ .

[00144] Step G. l-(6-Chloro-l-methyl-lH-indazol-3-yl)ethanone. To a solution of l-(6-chloro- lH-indazol-3-yl)ethanone (390 mg, 2 mmol) in DMF (5 mL) at 0 °C were added K ₂ C0 ₃ (552 mg, 4 mmol) and Mel (312 mg, 2.2 mmol). The resulting mixture was stirred at r.t for 4 hr, then poured into ice- water with vigorous stirring and extracted with EtOAc. The combined organic layers were washed with aq. LiCl (10%wt) and concentrated under reduced pressure. The residue was purified by flash column chromatography to give l-(6-chloro-l -methyl- lH-indazol-3-yl)ethanone (320 mg, 78.4% yield) as a white solid. LC-MS: m/z 209(M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 8.29 (d, = 8.4 Hz, 1H), 7.46 (d, = 1.2 Hz, 1H), 7.28-7.31 (m, 1H), 4.13 (s, 3H), 2.71 (s, 3H).

[00145] Step H. 2-bromo-l-(6-chloro-l-methyl-lH-indazol-3-yl)ethanone. To a solution of l-(6- chloro-l-methyl-lH-indazol-3-yl)ethanone (104 mg, 0.5 mmol) in EtOAc (5 mL) was added CuBr ₂ (245 mg, 1.1 mmol). The reaction mixture was stirred at reflux for 2 hr, then cooled to r.t. and filtered through Celite. The filtrate was washed with a solution of aq. Na ₂ S ₂ 0 ₃ and brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give crude2-bromo-l-(6-chloro-l-methyl-lH-indazol-3- yl)ethanone (160 mg, quant, crude yield) as pale yellow solid, which was used directly in the next step without any further purification. LC-MS: m/z287(M+H) ⁺ .

[00146] Step I. l-(6-Chloro-l-methyl-lH-indazol-3-yl)-2-(5-chloro-lH-benzo[d ]imidazol-2- ytihiojethanone (Compound 5) To a solution of the above crude 2-bromo-l-(6-chloro-l -methyl- 1H- indazol-3-yl)ethanone (145 mg, 0.5 mmol) in DMF (5 mL) were added 5-chloro-lH-benzo[d]imidazole- 2-thiol (110 mg, 0.6 mmol) and K ₂ C0 ₃ (138 mg, 1 mmol). The resulting mixture was stirred at r.t. for 2 hr, then poured into ice water and extracted with DCM. Combined organic layers were washed with aq. LiCl (10 wt), dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by prep-HPLC to give l-(6-chloro-l-methyl-lH-indazol-3-yl)-2-(5-chloro-lH-benzo[d ]imidazol- 2-ylthio)ethanone (60 mg, 30.6% yield) as white solid.

[00147] LC-MS: m/z: 391 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.86 (m, 1H), 8.09-8.13 (m, 2H), 7.37-7.50 (m, 3H), 7.09-7.15 (m, 1H), 5.07 (s, 2H), 4.22 (s, 3H).

EXAMPLE 4. SYNTHESIS OF 2-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YLTHIO)-1-(6-(PYRIDIN-3- -A]PYRIDIN-1-YL)ETHANONE (Compound 6)

[00148] Step A. 5-(Pyridin-3-yloxy)picolinonitrile. To a mixture of 5-fluoropicolinonitrile (1.22g, 10 mmol) and pyridin-3-ol (0.95 g, 10 mmol) in DMF (20 mL) was added K ₂ C0 ₃ (2.07 g, 15 mmol). The reaction mixture was stirred at r.t. for 2 hr, then quenched with water (10 mL) and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 5-(pyridin-3- yloxy)picolinonitrile (1.6 g, 84% yield) as light yellow oil. LC-MS: m/z 198(M+H) ⁺ .

[00149] Step B. (5-(Pyridin-3-yloxy)pyridin-2-yl)methanamine. To a solution of 5-(pyridin-3- yloxy)picolinonitrile (1.0 g, 5 mmol) in THF (30 mL) at 0 °C was added LiAlH ₄ (380 mg, 10 mmol). The reaction mixture was stirred at r.t. for 2 hr, then quenched with aq. NaOH (20% wt) and EtOAc. The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give (5-(pyridin-3-yloxy)pyridin-2-yl)methanamine (800 mg, 80% yield) as brown oil which was used in the next step without any further purification. LC-MS: m/z 202(M+H) ⁺ .

[00150] Step C. N-((5-(pyridin-3-yloxy)pyridin-2-yl)methyl)formamide. Amixture of (5-(pyridin- 3-yloxy)pyridin-2-yl)methanamine (600 mg, 3 mmol) in formic acid (6 mL) was heated at 110 °C for 12 hr, then cooled to 0 °C, quenched with aq. NH ₄ OH (25%wt) and extracted with DCM. The combined organic layers were dried over anhydrousNa ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography to give N-((5-(pyridin-3-yloxy)pyridin-2- yl)methyl)formamide (460 mg, 67% yield) as black oil. LC-MS: m/z 230(M+H) ⁺ .

[00\5l] Step D.6-(Pyridin-3-yloxy)imidazo[l,5-a]pyridine. To a mixture of N-((5-(pyridin-3- yloxy)pyridin-2-yl)methyl)formamide (460 mg, 2 mmol) in toluene (10 mL) at 0 °C was added POCl ₃ (612 mg,4 mmol). The reaction mixture was stirred at 100 °C for 12 hr, then cooled to 0 °C, quenched with aq. NH ₄ OH (25%wt) to pH9 and extracted with DCM. The combined organic layers were dried over anhydrous Na ₂ S0 and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-(pyridin-3-yloxy)imidazo[l,5-a]pyridine (210 mg, 50% yield) as black oil. LC-MS: m/z 212(M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 8.46 (d, = 2.9 Hz, 1H), 8.42 - 8.37 (m, 1H), 8.30 (dd, = 8.1, 7.2 Hz, 2H), 7.66 (d, = 9.7 Hz, 1H), 7.53 (m, 1H), 7.51 - 7.40 (m, 2H), 6.73 (m, 1H).

[00152] Step E. 2-chloro-l-(6-(pyridin-3-yloxy)imidazo[l,5-a]pyridin-l-yl)et hanone. To a mixture of 6-(pyridin-3-yloxy)imidazo[l,5-a]pyridine (210 mg, 1 mmol) and A1C1 ₃ (801 mg, 6 mmol) in CS ₂ (10 mL) at 0 °C was added chloroacetyl chloride (678 mg, 6 mmol). The reaction mixture was stirred at 60 °C for 2 hr, then cooled to 0 °C, quenched with aq.NaOAc (1 N) to pH 9, and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ and under reduced pressure. The residue was purified by flash column chromatography to give 2-chloro-l-(6-(pyridin-3-yloxy)imidazo[l,5- a]pyridin-l-yl)ethanone (140 mg, 50% yield) as white solid. LC-MS: m/z 288(M+H) ⁺ .

[00153] Step F. 2-(5-chloro-lh-benzo[d]imMazol-2-ylthio)-l-(6-(pyridin-3-ylo xy)imidazo[l,5- a]pyridin-l-yl)ethanone. A mixture of 2-chloro-l-(6-(pyridin-3-yloxy)imidazo[l,5-a]pyridin-l- yl)ethanone (57 mg, 0.2 mmol), 5-chloro-lH-benzo[d]imidazole-2-thiol (37 mg, 0.2 mmol), and potassium carbonate (55 mg, 0.4 mmol) in DMF was stirred at r.t. for 2 hr, then quenched with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford the desired product (51 mg, 58.6% yield) as white solid. LC-MS: m/z 436(M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.80 (d, = 11.3 Hz, 1H), 8.58 - 8.49 (m, 3H), 8.45 (dd, = 4.6, 1.2 Hz, 1H), 8.17 (d, = 9.7 Hz, 1H), 7.63 (m, 1H), 7.55 - 7.45 (m, 2H), 7.44 - 7.35 (m, 2H), 7.13 (t, = 7.6 Hz, 1H), 5.02 (s, 2H).

EXAMPLE 5. SYNTHESIS OF 2-((5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YL)THIO)-1-(6-(PYRIDIN-3- YLOXY)- -IND AZOL-3 - YL)ETH ANONE (Compound 7)

[00154] Step A. 2-Fluoro-4-(pyridin-3-yloxy)benzaldehyde. To a solution of 2,4- difluorobenzaldehyde (2.8 g, 20 mmol) in DMSO (40 mL) at r.t. were added K ₂ C0 ₃ (3.3 g, 24 mmol) and pyridin-3-ol (1.9 g, 20 mmol). The reaction mixture was stirred at r.t. overnight, then slowly poured into ice -water with vigorous stirring and extracted with EtOAc. The combined organic layers were washed with brine twice, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 2-fluoro-4-(pyridin-3-yloxy)benzaldehyde (3.1 g, 71.4% yield) as pale-yellow solid. LC-MS: m/z 218(M+H) ⁺ .

[00155] Step B. 6-(Pyridin-3-yloxy)-lH-indazole. To a solution of 2-fluoro-4-(pyridin-3- yloxy)benzaldehyde (2.17 g, 10 mmol) in dioxane (5 mL) in a sealed tube at 10 °C was added dropwise N ₂ H ₄ .H ₂ 0 (5 mL, 99%wt). After addition, the reaction mixture was stirred at r.t. for 0.5 hr, then stirred 120 °C for 48 hr. The resulting mixture was cooled to r.t. and concentrated under reduced pressure. The residue was partitioned between water and EtOAc. The organic layer was separated, washed with brine, and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-(pyridin-3-yloxy)-lH-indazole (1.2 g, 56.8% yield) as yellow syrup. LC-MS: m/z 212 (M+H) ⁺ .

[00156] Step C. 3-Iodo-6-(pyridin-3-yloxy)-lH-indazole.To a mixture of 6-(pyridin-3-yloxy)-lH- indazole (1.05 g, 5 mmol) in DMF (10 mL) at r.t. was added I ₂ (1.9 g, 7.5 mmol), followed by addition of well-ground KOH (1.12 g, 20 mmol). The reaction mixture was stirred at r.t., then poured into cooled S. aq. Na ₂ S ₂ 0 ₃ and extracted with EtOAc. The combined organic layers were washed with aq. LiCl (10% wt), dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 3-iodo-6-(pyridin-3-yloxy)-lH-indazole (1.3 g, 77.4% yield) as white solid. LC-MS: m/z 338 (M+H) ⁺ .

[00157] Step D. Methyl 6-(pyridin-3-yloxy)-lH-indazole-3-carboxylate. A mixture of 3-iodo-6- (pyridin-3-yloxy)-lH-indazole (1.1 g, 3.33 mmol), Pd(dppf)Cl ₂ (81.6 mg, 0.1 mmol), dppf (77 mg, 0.14 mmol), and Et ₃ N (0.57 g, 5.67 mmol) in a mixed solvent of toluene (15 mL) and MeOH (15 mL) was stirred at 75 °C under an atmosphere of CO (3 bar pressure) for 12 hr. The resulting mixture was cooled to r.t. and concentrated under reduced pressure. The residue was purified by flash column

chromatography to give methyl 6-(pyridin-3-yloxy)-lH-indazole-3-carboxylate (0.6 g, 66.9% yield) as gray solid. LC-MS: m/z 270 (M+H) ⁺ .

[00158] Step E. 6-(Pyridin-3-yloxy)-lH-indazole-3-carboxylic acid. To a solution of methyl 6- (pyridin-3-yloxy)-lH-indazole-3-carboxylate (540 mg, 2 mmol) in MeOH (30 mL) at r.t. were added THF (10 mL) and a solution of NaOH (400 mg, 10 mmol) in H ₂ 0 (10 mL). The reaction mixture was stirred at reflux for 5 hr, then cooled to r.t. and concentrated under reduced pressure to a volume of about 10 mL. The resulting mixture was acidified to pH 5-6 with cone. HC1 at 0 °C. The solid was collected by filtration, washed with H ₂ 0 and dried in high vacuum to give 6-(pyridin-3-yloxy)-lH-indazole-3- carboxylic acid (500 mg, 97.5% yield) as gray solid. LC-MS: m/z 256 (M+H) ⁺ .

[00159] Step F. N-methoxy-N-methyl-6-(pyridin-3-yloxy)-lH-indazole-3-carboxa mide. To a mixture of 6-(pyridin-3-yloxy)-lH-indazole-3-carboxylic acid (500 mg, 2 mmol) in THF (30 mL) at r.t. was added N,0-dimethylhydroxylamine hydrochloride (233 mg, 2.4 mmol), pyridine (821 mg, 10.4 mmol) and EDCI (691 mg, 3.6 mmol). The reaction mixture was stirred at r.t. for 16 hr, then poured into water and extracted with EtOAc. The combined organic layers were washed with brine twice, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give crude N-methoxy-N-methyl-6- (pyridin-3-yloxy)-lH-indazole-3-carboxamide (600 mg, 100% yield) as gray solid, which was directly used in the next step without any further purification. LC-MS: m/z 299 (M+H) ⁺ . [00160] Step G. l-(6-(Pyridin-3-yloxy)-lH-indazol-3-yl)ethanone. To a solution of the above crude N-methoxy-N-methyl-6-(pyridin-3-yloxy)-lH-indazole-3-carboxa mide (600 mg, 2 mmol) in dry THF (25 mL) at -5°C was dropwise added MeMgBr (3.33 mL, 10 mmol, 3 M in THF) over 10 min. After addition, the mixture was stirred at 0 °C for 1 hr, then slowly poured into an ice -cooled solution of aq. NH ₄ C1 with vigorous stirring and extracted with EtOAc. The combined organic layers were concentrated under reduced pressure, and the residue was purified by flash column chromatography to give l-(6- (pyridin-3-yloxy)-lH-indazol-3-yl)ethanone (320 mg, 64% yield) as white solid. LC-MS: m/z

254(M+H) ⁺ .

[00161] Step H. 2-Bromo-l-(6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone hydrobromide. To a solution of l-(6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone (50 mg, 0.2 mmol) in aq. HBr (2 mL, 48 wt) at r.t. was added a solution of Br ₂ (42 mg, 0.26 mmol) in aq. HBr (1 mL, 48 wt). The reaction mixture was stirred at r.t overnight and then concentrated under reduced pressure to dryness below 60 °C by an oil pump to give crude 2-bromo-l-(6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone hydrobromide salt (80 mg, 100% yield) as yellow solid which was directly used in the next step without any further purification. LC-MS: m/z 332(M+H) ⁺ .

[00162] Step I. 2-((5-chloro-lH-benzo[d]imidazol-2-yl)thio)-l-(6-(pyrMin-3-y loxy)-lH-indazol- 3-yl)ethanone. To a mixture of the above crude 2-bromo-l-(6-(pyridin-3-yloxy)-lH-indazol-3- yl)ethanone hydrobromide salt (66 mg, 0.2 mmol) in DMF (4 mL) were added 5-chloro-lH- benzo[d]imidazole-2-thiol (37 mg, 0.2 mmol) and K ₂ C0 ₃ (276 mg, 2 mmol). The reaction mixture was stirred at r.t. for 2 hr, then poured into ice water and extracted with DCM. The combined organic layers were washed with aq. LiCl (10%wt)and concentrated under reduced pressure. The residue was purified by prep- HPLC to give 2-(5-chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6-(pyridin-3-yl oxy)-lH-indazol-3- yl)ethanone (9 mg, 10.3% yield) as white solid. LC-MS: m/z 436 (M+H) ⁺ . H NMR (400 MHz, DMSO- d ₆ ) δ 8.47 (d, = 2.4 Hz, 1H), 8.42 (d, = 3.6 Hz, 1H), 7.40 - 7.56 (m, 4H), 7.25 (d, = 2.0 Hz, 1H), 7.11 - 7.16 (m, 2H), 5.11 (s, 2H).

EXAMPLE 6. SYNTHESIS OF 2-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YLTHIO)-1-(1-METHYL-6-(PYR IDIN- 3- YLOXY)- 1 H-IND AZOL-3 - YL)ETHANONE (Compound 8)

[00163] Step A. l-(l-Methyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone. To a solution of 1- (6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone (254 mg, 1 mmol) in DMF (5 mL) at r.t. were added K ₂ C0 ₃ (276 mg, 2 mmol) and Mel (156 mg, 1.1 mmol). The reaction mixture was stirred at r.t. overnight, then poured into ice -water with vigorous stirring and extracted with DCM. The combined organic layers were washed with aq. LiCl (10 wt), and concentrated under reduced pressure. The residue was purified by flash column chromatography to give l-(l-methyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone (170 mg, 63.4% yield) as white solid. LC-MS: m/z 268(M+H) ⁺ .

[00164] Step B. 2-Bromo-l-(l-methyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)etha none hydrobromide. To a solution of l-(l-methyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone (54 mg, 0.2 mmol) in aq. HBr (2 mL, 48 wt) at r.t. was added a solution of Br ₂ (42 mg, 0.26 mmol) in aq. HBr (1 mL, 48%). The reaction mixture was stirred at r.t. overnight and concentrated to dryness under reduced pressure below 60 °C by oil pump to give crude 2-bromo-l-(l-methyl-6-(pyridin-3-yloxy)-lH-indazol-3- yl)ethanone hydrobromide salt (80 mg, quant, crude yield) as yellow solid which was used directly in the next step without any further purification. LC-MS: m/z 346(M+H) ⁺ .

[00165] Step C. 2-(5-Chloro-lH-benzo[d]imidazol-2-ylthio)-l-(l-methyl-6-^yri din-3-yloxy)- lH-indazol-3-yl)ethanone . To a mixture of the above crude 2-bromo-l-(l-methyl-6-(pyridin-3-yloxy)- lH-indazol-3-yl)ethanone hydrobromide salt (80 mg, 0.2 mmol) in DMF (4 mL) were added 5-chloro- lH-benzo[d]imidazole-2-thiol (56 mg, 0.3 mmol) and K ₂ C0 ₃ (276 mg, 2 mmol). The reaction mixture was stirred at r.t. for 2 hr, then poured into ice water and extracted with DCM. The combined organic layers were washed with aq. LiCl (10%wt)and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 2-(5-chloro-lH-benzo[d]imidazol-2-ylthio)-l-(l-methyl-6-(pyr idin-3- yloxy)-lH-indazol-3-yl)ethanone (13 mg, 14.4% yield) as white solid. LC-MS: m/z 450 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.86 (br, 1H), 8.41 - 8.47 (m, 2H), 8.14 (d, = 8.4 Hz, 1H), 7.41 - 7.54 (m, 5H), 7.12 - 7.18 (m, 2H), 5.08 (s, 2H), 4.16 (s, 3H). EXAMPLE 7. SYNTHESIS OF OF 2-((5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YL)THIO)-1-(6-(PYRIDIN-3- YLOX Y)- -INDOL-3 - YL)ETHANONE (Compound 9)

[00166] Step A. 2-Chloro-l-(6-(pyridin-3-yloxy)-lH-indol-3-yl)ethanone. To a stirred solution of 6-(pyridin-3-yloxy)-lH-indole (200 mg, 0.95 mmol) in anhydrous DCM (10 mL) at 0 °C was added A1C1 ₃ (126 mg, 0.95 mmol), followed by addition of 2-chloroacetyl chloride (0.06 mL, 0.76 mmol). The reaction mixture was slowly warmed to r.t and stirred for 16 hr. The resulting mixture was quenched with water, and extracted with DCM (20 mL). The combined organic layers were washed with brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by column chromatography (PE/EtOAc = 1/1) to afford 2-chloro-l-(6-(pyridin-3-yloxy)-lH-indol-3-yl)ethanone (50 mg, 18.5% yield) as yellow solid. LC-MS: m z 287 (M+H) ⁺ .

[00167] Step B. 2-(5-Chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6-(pyridin-3-yl oxy)-lH-indol-3- yljethanone. To a solution of 2-chloro-l-(6-(pyridin-3-yloxy)-lH-indol-3-yl)ethanone (45 mg, 0.16 mmol) in DMF (3 mL) at r.t. were added 5-chloro-lH-benzo[d]imidazole-2-thiol (30 mg, 0.16 mmol) and K ₂ C0 ₃ (66 mg, 0.48 mmol). The resulting mixture was stirred at room temperature for 2 h, then quenched with H ₂ 0 and extracted with EtOAc. The combined organic layers were washed with aq. LiCl (10% wt) and brine, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford 2-(5-chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6-(pyridin-3-yl oxy)-lH- indol-3-yl)ethanone (35 mg, 51.5% yield) as white solid.

[00168] LC-MS: m z 435 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.83 (s, 1H), 12.08 (s, 1H), 8.56 (s, 1H), 8.39 (s, 1H), 8.37 - 8.32 (m, 1H), 8.15 (d, = 8.6 Hz, 1H), 7.53 - 7.38 (m, 4H), 7.19 - 7.11 (m, 2H), 7.01 (dd, = 8.6, 2.2 Hz, 1H), 4.88 (s, 2H).

[00169] The procedure set forth above was used to produce the following compounds using the appropriate starting materials.

[()()\l()]2-(5-chloro-lH-benzo[d]imidazol-2-ylthio)-l-(l-met hyl-6-(pyri^

yl)ethanone (Compound 10)

[00171] LC-MS: m/z 449 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.85 (s, 1H), 8.60 (s, 1H), 8.37 (d, = 17.1 Hz, 2H), 8.17 (s, 1H), 7.40 (s, 5H), 7.15-7.05 (m, 2H), 4.86 (s, 2H), 3.86 (s, 3H).

EXAMPLE 8. SYNTHESIS OF 3-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YL)-1-(1-METHYL-6-(PYRIDIN -3- -lH-rNDOL-3-YL)PROPAN-l-ONE (Compound 11)

[00172] Step A. Ethyl 4-(l-methyl-6-(pyridin-3-yloxy)-lH-indol-3-yl)-4-oxobutanoat e. To a mixture of l-methyl-6-(pyridin-3-yloxy)-lH-indole (300 mg, 1.3 mmol) in anhydrous DCM (5 mL) at 0 °C was added A1C1 ₃ (621 mg, 2.6 mmol), followed by addition of a solution of ethyl 4-chloro-4- oxobutanoate (0.37 mL, 2.6 mmol) in anhydrous DCM (1 mL). The mixture was stirred at 0 °C for lhr, then quenched with aq. HCl (0.1 N, 10 drops)and extracted with DCM. The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by column chromatography to afford the desired product (100 mg, 22% yield) as yellow oil. LC-MS: m/z 353 (M+H) ⁺ .

[00173] Step B. 4-(l-Methyl-6-(pyridin-3-yloxy)-lH-indol-3-yl)-4-oxobutanoic acid. To a mixture of ethyl 4-(l-methyl-6-(pyridin-3-yloxy)-lH-indol-3-yl)-4-oxobutanoat e (100 mg, 0.28 mmol) in THF (2 mL) was added a solution of NaOH (34 mg, 0.85 mmol) in H ₂ 0 (2 mL). The reaction mixture was stirred at r.t. for 2 hr, then adjusted to pH 6 with aq. HCl (IN) and extracted with EtOAc. The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by prep-TLC to afford the desired product (35 mg, 39% yield) as yellow solid. LC-MS: m/z 325 (M+H) ⁺ .

[00174] Step C. N-(2-amino-4-chlorophenyl)-4-(l-methyl-6-(pyridin-3-yloxy )-lH-indol-3-yl)-4- oxobutanamide and N-(2-amino-5-chlorophenyl)-4-(l-methyl-6-(pyridin-3-yloxy)-l H-indol-3-yl)-4- oxobutanamide. To a mixture of 4-chlorobenzene-l,2-diamine (16 mg, 0.11 mmol), 4-(l-methyl-6- (pyridin-3-yloxy)-lH-indol-3-yl)-4-oxobutanoic acid (35 mg, 0.1 mmol), EDCI (23 mg, 0.12 mmol), and HOBT (27 mg, 0.2 mmol) in DMF (2 mL) was added DIPEA (0.05 mL, 0.3 mmol). The reaction mixture was stirred at r.t. overnight, then diluted with water and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give the crude product (43 mg, 96% yield) as brown oil. LC-MS: m/z 449(M+H) ⁺ .

[00175] Step D. 3-(5-chloro-lH-benzo[d]imidazol-2-yl)-l-(l-methyl-6-(pyridin -3-yloxy)-lH- indol-3-yl)propan-l-one. A mixture of N-(2-amino-4-chlorophenyl)-4-(l-methyl-6-(pyridin-3-yloxy)-l H- indol-3-yl)-4-oxobutanamide and N-(2-amino-4-chlorophenyl)-5-(l-methyl-6-(pyridin-3-yloxy)-l H- indol-3-yl)-4-oxobutanamide (43 mg, 0.10 mmol) in AcOH (2 mL) was stirred at reflux for 2 hr, then cooled to r.t. and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford 3-(5-chloro-lH-benzo[d]imidazol-2-yl)-l-(l-methyl-6-(pyridin -3-yloxy)-lH-indol-3-yl)propan-l-one (10 mg, 23% yield) as white solid. LC-MS: m/z 431 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.46 (d, = 16.2 Hz, 1H), 8.45 (s, 1H), 8.40 - 8.28 (m, 2H), 8.18 (d, = 8.6 Hz, 1H), 7.57 - 7.32 (m, 5H), 7.13 (s, 1H), 7.00 (d, = 7.8 Hz, 1H), 3.82 (s, 3H), 3.45 (d, = 7.3 Hz, 2H), 3.18 (t, = 6.9 Hz, 2H).

EXAMPLE 9. SYNTHESIS OF 2-((5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YL)THIO)-1-(7-(PYRIDIN-3- -A]PYRIDIN-3-YL)ETHANONE (Compound 12)

[00176] Step A. 4-Chloro-2-(2,5-dimethyl-lH-pyrrol-l-yl)pyridine. To a mixture of 4- chloropyridin-2-amine (2.56 g, 20 mmol) in toluene (50 mL) were added hexane-2,5-dione (2.7 g, 24 mmol) and TPSA (0.1 g). The reaction mixture was stirred at reflux for 12 hr, then cooled to r.t. and diluted with EtOAc. The resulting mixture was washed with water twice, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 4-chloro-2-(2,5 -dimethyl- lH-pyrrol-l-yl)pyridine (2.6 g, 63% yield) as yellow oil. LC-MS: m/z 207(M+H) ⁺ .

[00177] Step B. 2-(2,5-Dimethyl-lH-pyrrol-l-yl)-4-(pyridin-3-yloxy)pyridine. To a solution of 4- chloro-2-(2,5-dimethyl-lH-pyrrol-l-yl)pyridine (2.06 g, 10 mmol) in DMF (30 mL) at 0 °C were added pyridin-3-ol (1.4 g, 15 mmol) and t-BuOK (2.2 g, 20 mmol). After addition, the mixture was slowly heated to 90 °C and stirred at that temperature for 16 hr, then cooled to r.t., poured into ice-water and extracted with EtOAc. The combined organic layers were washed with aq. LiCl (10%wt), dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 2-(2,5-dimethyl-lH-pyrrol-l-yl)-4-(pyridin-3-yloxy)pyridine (1.7 g, 64.1% yield) as yellow oil. LC-MS: m/z 266 (M+H) ⁺ .

[00178] Step C. 4-(Pyridin-3-yloxy)pyridin-2-amine. To a mixture of 2-(2,5-dimethyl-lH-pyrrol- l-yl)-4-(pyridin-3-yloxy)pyridine (1.32 g, 5 mmol) at 0 °C in MeOH (50 mL) was added cone. HC1 (10 mL). The resulting solution was stirred reflux for 16 hr, then concentrated under reduced pressure to give crude 4-(pyridin-3-yloxy)pyridin-2-amine HC1 salt (1.2 g, quant, crude yield) as yellow solid which was used directly in the next step without any further purification. LC-MS: m/z 188 (M+H) ⁺ .

[00179] Step D. 7-(Pyridin-3-yloxy)imidazo[l,2-a]pyridine. To a mixture of the above crude 4- (pyridin-3-yloxy)pyridin-2-amine (1.2 g, 5 mmol) in DCM (30 mL) were added saturated aq. NaHC0 ₃ (30 mL) and 2-chloroacetaldehyde (2.4 g, 12.3 mmol, 40%wt in water). The reaction was stirred vigorously at r.t. for 16 hr. The organic layer was separated, and the aq. layer was extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 7-(pyridin-3- yloxy)imidazo[l,2-a]pyridine (0.7 g, 66.3% yield) as gray solid. LC-MS: m/z 212 (M+H) ⁺ .

[00180] S¾77 E.2-Chloro-l-(7-(pyridin-3-yloxy)imidazo[l,2-a]pyridin-3-yl) ethanone. To a mixture of 7-(pyridin-3-yloxy)imidazo[l,2-a]pyridine (210 mg, 1 mmol), A1C1 ₃ (801 mg, 6 mmol) in CS ₂ (10 mL)in an ice bath was added chloroacetyl chloride (678 mg, 6 mmol). The reaction mixture was stirred at 60 °C for 2 hr, then cooled to 0 °C, basified with aq. sodium acetate (IN) and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 2-chloro-l-(7-(pyridin-3- yloxy)imidazo[l,2-a]pyridin-3-yl)ethanone (140 mg, 50% yield) as white solid. LC-MS: m/z 288(M+H) ⁺ . [00181] Step F. 2-((5-chloro-lH-benzo[d]imidazol-2-yl)thio)-l-(7-(pyrMin-3-y loxy)imid a]pyridin-3-yl)ethanone. A mixture of 2-chloro-l-(7-(pyridin-3-yloxy)imidazo[l,2-a]pyridin-3- yl)ethanone (57 mg, 0.2 mmol), 5-chloro-lH-benzo[d]imidazole-2-thiol (37 mg, 0.2 mmol), and potassium carbonate (55 mg, 0.4 mmol) in DMF was stirred at r.t. for 2 hr, then quenched with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford the desired product (51 mg, 58.6% yield) as white solid. LC-MS: m/z 436(M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.81 (s, 1H), 9.44 (d, = 7.5 Hz, 1H), 8.78 (s, 1H), 8.55 (dd, / = 15.4, 3.6 Hz, 2H), 7.74 (dd, = 8.4, 1.5 Hz, 1H), 7.61 - 7.34 (m, 3H), 7.28 - 7.07 (m, 3H), 4.88 (d, = 13.2 Hz, 2H).

EXAMPLE 10. SYNTHESIS OF PREPARATION OF 2-(5-CHLORO-7-(2-HYDROXYPROPAN-2-YL)-1H- -2-YLTHIO)-l-(6-(PYRIDTN-3-YLOXY)-lH-INDOL-3-YL)ETHANONE (Compound 13)

[00182] Step A: 2-chloro-l-(6-(pyridin-3-yloxy)-lH-indol-3-yl)ethanone. To a mixture of 6- (pyridin-3-yloxy)-lH-indole (200 mg, 0.95 mmol) in anhydrous DCM (3 mL) at 0 °C was added A1C1 ₃ (190 mg, 1.43 mmol), followed by addition of a solution of 2-chloroacetyl chloride (0.11 mL, 1.43 mmol) in anhydrous DCM (1 mL). The mixture was stirred at 0 °C for 3 hr, then quenched with aq. HCl (0.1 N, 10 drops) and partitioned between DCM and H ₂ 0. The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by column chromatography to afford the desired product (75 mg, 28% yield) as yellow solid. LC-MS: m/z 287(M+H) ⁺ .

[00183] Step B. 2-(5-Chloro-7-(2-hydroxypropan-2-yl)-lH-benzo[d]imidazol-2-y lthio)-l-(6- (pyridin-3-yloxy)-lH-indol-3-yl)ethanone. To a mixture of 2-chloro-l-(6-(pyridin-3-yloxy)-lH-indol-3- yl)ethanone (40 mg, 0.14 mmol) in DMF (3 mL) were added 2-(5-chloro-2-mercapto-lH- benzo[d]imidazol-7-yl)propan-2-ol (34 mg, 0.14 mmol) and K ₂ C0 ₃ (58 mg, 0.42 mmol). The reaction mixture was stirred at r.t. for 2 hr, then partitioned between EtOAc and H ₂ 0. The organic layer was washed with aq. LiCl (10 wt) and brine, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by flash column chromatography to afford the desired product (25 mg, 36% yield) as yellow solid. LC-MS: m/z 493 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.75-12.31 (m, 1H), 12.08 (s, 1H), 8.57 (s, 1H), 8.42 - 8.30 (m, 2H), 8.15 (d, = 8.6 Hz, 1H), 7.45 - 7.25 (m, 3H), 7.21 - 6.96 (m, 3H), 5.46-5.14 (m, 1H), 4.75 (s, 2H), 1.54-1.38 (m, 6H).

EXAMPLE 11. SYNTHESIS OF 2-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YLTHIO)-1-(1-ETHYL-6-(PYRI DIN-3- - 1 H-INDOL-3 - YL)ETHANONE (Compound 14)

[00184] Step A: l-ethyl-6-(pyridin-3-yloxy)-lH-indole. To a suspension of 6-(pyridin-3-yloxy)- lH-indole (400 mg, 1.9 mmol) in DMF (20 mL) at 0 °C was added NaH (114 mg, 4.75 mmol). The reaction mixture was stirred at 0 °C for 10 min, followed by slow addition of iodoethane (371 mg, 2.38 mmol). The resulting mixture was slowly warmed to r.t. over 1 hr, then quenched with water, and extracted with EtOAc (20 mL). The organic layer was separated, washed in sequence with water (2x10 mL) and brine (10 mL), dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by column chromatography (PE/EtOAc = 5: 1) to afford the desired product (400 mg, 88.5% yield) as yellow oil. LC-MS: m/z 239 (M+H) ⁺ .

[00185] Ste/7 B: 2-Chloro-l-(l-ethyl-6-(pyridin-3-yloxy)-lH-indol-3-yl)ethano ne. To a stirred solution of l-ethyl-6-(pyridin-3-yloxy)-lH-indole (150 mg, 0.63 mmol) in anhydrous DCM (10 mL) at 0 °C was added A1C1 ₃ (126 mg, 0.95 mmol), followed by dropwise addition of a solution of 2-chloroacetyl chloride (107 mg, 0.95 mmol) in anhydrous DCM (2 mL). The resulting mixture was stirred at r.t. for 16 hr, then quenched with water and extracted with DCM (20 mL).The organic layer was separated, washed with brine (10 mL), dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give the desired product (120 mg, 60.7% yield) as yellow solid. LC-MS: m/z 315 (M+H) ⁺ .

[00186] Step C: 2-(5-Chloro-lH-benzo[d]imMazol-2-ylthio)-l-(l-ethyl-6-(pyrid in-3-yloxy)-lH- indol-3-yl)ethanone . To a mixture of 2-chloro-l-(l-ethyl-6-(pyridin-3-yloxy)-lH-indol-3-yl)ethano ne (40 mg, 0.13 mmol) in DMF (1 mL) at r.t. were added 5-chloro-lH-benzo[d]imidazole-2-thiol (23 mg, 0.13 mmol) and K ₂ C0 ₃ (54 mg, 0.39 mmol). The resulting mixture was stirred at r.t. for 1 hr then filtrated. The filtrate was purified by prep-HPLC to afford the desired product (28.4 mg, 47.3% yield) as white solid. LC-MS: m/z 463 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.83 (s, 1H), 8.56 (s, 1H), 8.38 (d, = 2.4 Hz, 1H), 8.35 - 8.31 (m, 1H), 8.17 (d, = 8.8 Hz, 1H), 7.50 - 7.30 (m, 5H), 7.14 (ddJ = 8.4 Hz, 1H), 7.03 (ddJ = 8.8 Hz, 1H), 4.86 (s, 2H), 4.30 - 4.20 (m, 2H), 1.40 (t, 3H).

[00187] The procedure set forth above was used to produce the following compounds using the appropriate starting materials.

[00188] 2-( 5-chloro-7-( 2-hydroxypropan-2-yl)-lH-benzo[ d]imidazol-2-ylthio)-l-(l-ethyl-6- (pyridi -3-yloxy)-lH-indol-3-yl)ethanone (Compound 15)

[00189] LC-MS: m/z 521 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.36 (s, 1H), 8.66 (s, 1H), 8.37 (d, = 2 Hz, 1H), 8.37 (d, = 3.2 Hz, 1H), 8.17 (d, = 8.8 Hz, 1H), 7.47 (d, = 1.6 Hz, 1H), 7.42- 7.30 (m, 3H), 7.14 (s, 1H), 7.01 (d,J = 7.6 Hz, 1H), 5.19-5.14 (m, 1H), 4.75 (s, 2H), 4.30 - 4.20 (m, 2H), 1.70-1.20 (m, 9H).

EXAMPLE 12. SYNTHESIS OF 2-(6-CHLORO-1H-BENZO[D]IMIDAZOL-2-YLTHIO)-1-(1-ETHYL-6-(PYRI DIN- - YLOXY)-lH-rNDAZOL-3-YL)ETHANONE (Compound 16)

[00190] Step A: l-(l-Ethyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone. A mixture of 1-(1- ethyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone (260 mg, 1.03 mmol), iodoethane (160 mg, 1.03 mmol), and K ₂ C0 ₃ (284 mg, 2.05 mmol) in DMF (5 mL) was stirred at r.t. for 1 hr, then diluted with EtOAc and washed with aq. LiCl (10 wt). The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give l-(l-ethyl-6-(pyridin-3-yloxy)-lH-indazol-3- yl)ethanone (220 mg, 76% crude yield) as yellow semi-solid which was used in the next step without any further purification.LC-MS: m/z 282 (M+H) ⁺ .

[00191] Step B: 2-Chloro-l-(l-ethyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)etha none. A mixture of l-(l-ethyl-6-(pyridin-3-yloxy)-lH-indazol-3-yl)ethanone (220 mg, 0.782 mmol), and TEA (103 mg, 1.02 mmol) in DCM (lOmL) was cooled to -5°C, followed by addition of TMSOTf (348 mg, 1.54 mmol). The mixture was stirred at that temperature for 2 hr, followed by addition of NCS (104 mg, 0.782 mmol) in one portion. The resulting mixture was stirred at that temperature for anther lhr, then diluted with EtOAc and washed with S. aq. NaHC0 ₃ . The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give 2-chloro-l-(l-ethyl-6-(pyridin-3- yloxy)-lH-indazol-3-yl)ethanone (90 mg, 63.45% yield) as yellow semi-solid which was used in the next step without any further purification.LC-MS: m/z 316 (M+H) ⁺ .

[00192] Step C: 2-(6-Chloro-lH-benzo[d]imidazol-2-ylthio)-l-(l-ethyl-6-(pyri din-3-yloxy)- 1H- indazol-3-yl)ethanone . To a mixture of 2-chloro-l-(l-ethyl-6-(pyridin-3-yloxy)-lH-indazol-3- yl)ethanone (45mg, 0.143 mmol), 5-chloro-lH-benzo[d]imidazole-2-thiol (26 mg, 0.143 mmol) in DMF (5 mL) was added K ₂ C0 ₃ (48 mg, 0.286 mmol). The reaction mixture was stirred at r.t. for 1 hr, then diluted with EtOAc and washed with aq. LiCl (10%wt). The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 2-(6 -Chloro- 1 H-benzo [d] imidazol-2-ylthio)- 1 -( 1 -ethyl-6 -(pyridin-3 -yloxy) - 1 H-indazol-3 - yl)ethanone (60 mg, 90.7% yield) as white solid.LC-MS: m/z 464(M+H) ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ )88.44 (d, = 2 Hz, 4H), 8.40 (dd, = 2, 4 Hz, 1H), 8.14 (d, = 8 Hz, 4H), 7.58 (d, = 2 Hz, 1H), 7.36 - 7.54 (m, 4H), 7.14 (d, = 2 Hz, 2H), 5.06 (s, 2H), 4.46 - 4.58 (m, 2H), 1.43 (s, 3H)

[00193] The procedure set forth above was used to produce the following compounds using the appropriate starting materials.

[00194] 2-( ( 6-Chloro-lH-benzo[ d]imidazol-2-yl)thio)-l-(l-( 2-hydroxy-2-methylpropyl)-6- (pyridin-3 d 17)

C [00195] LC-MS: m/z 508 (M+H)VH NMR (400 MHz, DMSO-d ₆ )88.44 (d, / = 2 Hz, 1H), 8.38 - 8.41 (m, 1H), 8.14 (d, =8 Hz, 1H), 7.60 (d, = 2 Hz, 5H), 7.47 (d, = 2 Hz, 4H), 7.11 - 7.18 (m, 2H), 5.06 (s, 2H), 4.70 - 4.82 (m, 1H), 4.41 (s, 2H), 1.17 (s, 6H).

EXAMPLE 13. SYNTHESIS OF 2-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YLTHIO)-1-(6-(PYRAZIN-2-YL OXY)- 1H-TNDOL-3-YL) E (Compound 18)

[00196] Step A: 6-(Pyrazin-2-yloxy)-lH-indole. A mixture of lH-indol-6-ol (300 mg, 2.25 mmol) and 2-chloropyrazine (284 mg, 2.48 mmol) and K ₂ C0 ₃ (1.47 g, 4.51 mmol) in DMF (10 mL) was stirred at 100 °C for 1 hr, then diluted with water and extracted with EtOAc. The combined organic layers were washed with S. aq. LiCl, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by flash column chromatography to afford 6-(pyrazin-2-yloxy)-lH- indole (260 mg, 55% yield) as yellow solid. LC-MS: m/z 212 (M+H)VH NMR (400 MHz, DMSO-d ₆ ) δ 11.15 (s, 1H), 8.49 (d, = 1.1 Hz, 1H), 8.34 (d, = 2.6 Hz, 1H), 8.18 (dd, = 2.5, 1.4 Hz, 1H), 7.57 (d, = 8.5 Hz, 1H), 7.37 (t, = 2.7 Hz, 1H), 7.22 (d, = 1.5 Hz, 1H), 6.84 (dd, = 8.5, 2.1 Hz, 1H), 6.46 (s, 1H).

[00197] Step B: 2-Chloro-l-(6-(pyrazin-2-yloxy)-lH-indol-3-yl)ethanone. A mixture of 6- (pyrazin-2-yloxy)-lH-indole (100 mg, 0.47 mmol) and A1C1 ₃ (315 mg, 2.37 mmol) in anhydrous DCM (5 mL) was stirred at 0 °C for 10 min, followed by slow addition of 2-chloroacetyl chloride (0.2 mL, 2.37 mmol). The reaction mixture was stirred at r.t. for 3 hr, then poured into ice-water and extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure to afford 2-chloro-l-(6-(pyrazin-2-yloxy)-lH-indol-3-yl)ethanone (55 mg, 40% crude yield) as a grey solid which was used in the next step without any further purification. LC-MS: m/z 288 (M+H) ⁺ . [00198] Step C: 2-(5-Chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6-(pyrazin-2-yl oxy)-lH-indol- 3-yl)ethanone. A mixture of 2-chloro-l-(6-(pyrazin-2-yloxy)-lH-indol-3-yl)ethanone (55 mg, 0.19 mmol) and 5-chloro-lH-benzo[d]imidazole-2-thiol (42 mg, 0.23 mmol), and K ₂ C0 ₃ (40 mg, 0.27 mmol) in DMF (5 mL) was stirred at r.t. for 1 hr, then diluted with water and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ , and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford 2-(5-chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6- (pyrazin-2-yloxy)-lH-indol-3-yl)ethanone (12 mg, 14% yield) as white solid. LC-MS: m/z 436(M+H)VH NMR (400 MHz, DMSO-d ₆ ) δ 12.81 (s, 1H), 12.18 (s, 1H), 8.59 (s, 1H), 8.54 (d, = 1.1 Hz, 1H), 8.36 (d, = 2.6 Hz, 1H), 8.19 (dd, / = 2.5, 1.3 Hz, 1H), 8.15 (d, = 8.6 Hz, 1H), 7.48 (s, 1H), 7.42 (d, = 8.5 Hz, 1H), 7.35 (d, = 2.0 Hz, 1H), 7.13 (dd, = 8.5, 1.9 Hz, 1H), 7.05 (dd, = 8.6, 2.1 Hz, 1H), 4.89 (s, 2H).

EXAMPLE 14. SYNTHESIS OF 2-(6-CHLORO-3H-IMIDAZO[4,5-C]PYRIDIN-2-YLTHIO)-1-(5-(PYRIDIN -3- -lH-rNDOL-3-YL)ETHANONE (Compound 19)

[00199] Step A: 6-Chloro-3H-imidazo[4,5-c]pyridine-2-thiol. To a mixture of 6- chloropyridine-3,4-diamine(286 mg, 2 mmol) in THF (10 mL) was added di-(lH-imidazol-l- yl)methanethione (534 mg,3 mmol). The reaction mixture was stirred at 80 °C for 4 hr, then diluted with water and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-chloro-3H-imidazo[4,5-c]pyridine-2-thiol (200 mg, 54 % yield).

[00200] Step B: 2-(6-Chloro-3H midazo[4,5-c]pyrMin-2-ylthio)-l-(5-(pyridin-3-yloxy)-lH- indol-3-yl)ethanone. To a mixture of 6-chloro-3H-imidazo[4,5-c]pyridine-2-thiol (184 mg,l mmol) in DMF (lOmL) were added K ₂ C0 ₃ (276 mg, 2 mmol), and 2-chloro-l-(5-(pyridin-3-yloxy)-lH-indol-3- yl)ethanone (286 mg, 1 mmol). The reaction mixture was stirred at r.t. for 1 hr, then diluted with water, and extracted with EtOAc. The combined organic layers were dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give 2-(6-chloro-3H-irmdazo[4,5-c]pyridin-2-ylthio)-l-(5-(pyridin -3-yloxy)-lH-indol-3- yl)ethanone (30 mg, 7% yield). LC-MS: m/z 436 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.10 (s, 1H), 8.57 (s, 1H), 8.50 (s, 1H), 8.39 (s, 1H), 8.37 - 8.32 (m, 1H), 8.14 (d, = 8.6 Hz, 1H), 7.51 (s, 1H), 7.41 (d, = 2.5 Hz, 2H), 7.17 (d, = 2.1 Hz, 1H), 7.01 (dd, = 8.6, 2.2 Hz, 1H), 4.93 (s, 2H).

[00201] The procedure set forth above was then used to produce 2-(5-Chloro-6-fluoro-lH- 0)

[00202] LC-MS: m/z: 454(M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 8.47 (d, = 2.8 Hz, 1H), 8.43 (dd, = 4.6, 1.3 Hz, 1H), 8.15 (d, = 8.8 Hz, 1H), 7.62 (d, = 6.8 Hz, 1H), 7.58 - 7.52 (m, 1H), 7.48 (dd, / = 8.8, 4.2 Hz, 2H), 7.25 (d, = 1.9 Hz, 1H), 7.15 (dd, / = 8.8, 2.1 Hz, 1H), 5.11 (s, 2H).

EXAMPLE 15. SYNTHESIS OF 3-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YL)-1-(6-(PYRIDIN- 3-YLOXY)-1H- -3-YL)PROPAN-1-ONE (Compound 21)

[00203] Step A. Methyl 4-oxo-4-(l-(phenylsulfonyl)-6-(pyridin-3-yloxy)-lH-indol-3-y l) butanoate. To a mixture of l-(phenylsulfonyl)-6-(pyridin-3-yloxy)-lH-indole (455 mg, 1.3 mmol) in anhydrous DCM (5 mL) at 0 °C was added A1C1 ₃ (621 mg, 2.6 mmol), followed by dropwise addition of a solution of methyl 4-chloro-4-oxobutanoate (0.37 mL, 2.6 mmol) in anhydrous DCM (1 mL). The reaction mixture was stirred at 0 °C for 1 hr, then quenched with aq. HCl (0.1 N, 10 drops) and extracted with DCM. The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by gel column chromatography to afford the desired product (140 mg, 22% yield) as yellow oil. LC-MS: m/z 465 (M+H) ⁺ .

[00204] Step B. 4-Oxo-4-(6-(pyridin-3-yloxy)-lH-indol-3-yl)butanoic acid. To a mixture of methyl 4-oxo-4-(l-(phenylsulfonyl)-6-(pyridin-3-yloxy)-lH-indol-3-y l)butanoate (120 mg, 0.28 mmol) in THF (2 mL) was added a solution of NaOH (34 mg, 0.85 mmol) in H ₂ 0 (2 mL). The reaction mixture was stirred at r.t. for 2 hr, then adjusted to pH 6 with aq. HCl (IN) and extracted with EtOAc. The organic layer was separated, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure. The residue was purified by prep-TLC to afford the desired product (50 mg, 55% yield) as yellow solid. LC-MS: m/z 311 (M+H) ⁺ .

[00205] Step C. N-(2-amino-4-chlorophenyl)-4-oxo-4-(6-(pyridin-3-yloxy)-lH-i ndol-3-yl) butanamide and N-(2-amino-5-chlorophenyl)-4-oxo-4-(6-(pyridin-3-yloxy)-lH-i ndol-3-yl) butanamide.

To a mixture of 4-chlorobenzene-l,2-diamine (30mg, O.l lmmol), 4-oxo-4-(6-(pyridin-3-yloxy)-lH-indol- 3-yl)butanoic acid (33 mg, 0.1 mmol), EDCI (23 mg, 0.12 mmol), and HOBT (27 mg, 0.2 mmol) in DMF (2 mL) was added DIPEA (0.05 mL, 0.3 mmol). The reaction mixture was stirred at r.t. overnight, then diluted with water and extracted with EtOAc. Combined organic layers were dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give the crude product (43 mg, 96% yield) as brown oil. LC-MS: m/z 435(M+H) ⁺ .

[00206] Step D. 3-(5-chloro-lH-benzo[d]imidazol-2-yl)-l-(6-(pyridin-3-yloxy) -lH-indol-3-yl) propan-l-one. A mixture of N-(2-amino-4-chlorophenyl)-4-oxo-4-(6-(pyridin-3- yloxy)-lH-indol-3- yl)butanamide (43 mg, 0.10 mmol) in AcOH (2 mL) was refluxed for 2 hr, then cooled down and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford 3-(5-chloro-lH- benzo[d]imidazol-2-yl)-l-(6- (pyridin-3-yloxy)-lH-indol-3-yl)propan-l-one (10 mg, 23% yield) as white solid. LC-MS: m/z 417 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.34 - 12.21 (m, 2H), 11.94 (s, 1H), 8.49 - 8.27 (m, 3H), 8.18 (d, = 8.6 Hz, 1H), 7.70 - 7.29 (m, 4H), 7.12 (d, = 2.0 Hz, 2H), 6.98 (dd, = 8.6, 2.1 Hz, 1H), 3.46 (d, J = 1.2 Hz, 2H), 3.18 (t, = 7.2 Hz, 2H).

EXAMPLE 16. SYNTHESIS OF 2-(5-CHLORO-1H-BENZO[D]IMIDAZOL-2-YLTHIO)-1-(6- (PYRIMIDIN-2- YLOX Y)- 1 H-INDOL-3 - YL)ETHANONE (Compound 22)

[00207] Step A. 2-(4-Methyl-3-nitrophenoxy)pyrimidine. To a mixture of 4-methyl-3-nitrophenol (3.1 g, 20.3 mmol) and 2-chloropyrimidine (3.46 g, 30.4 mmol) in DMF (20 mL) was added Cs ₂ C0 ₃ (16.5 g, 50.75 mmol). The reaction mixture was stirred at 100°C for 1 hr then concentrated under reduced pressure. The residue was purified by column chromatography (PE:EtOAc = 2: 1) to afford the desired product (3.75 g, 80.0% yield) as white solid. LC-MS: m/z 232 (M+H) ⁺ .

[00208] Step B. (E)-2-(3-nitro-4-(2-(pyrrolidin-l-yl)vinyl)phenoxy)pyrimidin e. To a solution of 2-(4-methyl-3-nitrophenoxy)pyrimidine (3 g, 13 mmol) in DMF (1 mL) were added pyrrolidine (1.29 g, 18.2 mmol) and DMF-DMA (dimethyl acetal) (2.16 g, 18.2 mmol). The reaction mixture was stirred at 65°C for 2 hr then concentrated under reduced pressure to give the crude product (4 g) as black oil which was directly used in the next step without any further purification. LC-MS: m/z 313 (M+H) ⁺ .

[00209] Step C. 6-(Pyrimidin-2-yloxy)-lH-indole. To a mixture of (E)-2-(3-nitro-4-(2- (pyrrolidin-l-yl)vinyl)phenoxy)pyrimidine (4 g) in EtOAc (20 mL) was added Pd/C (10 wt, 0.4 g). The reaction mixture was stirred at r.t. under H ₂ atmosphere overnight then filtered. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography

(PE:EtOAc = 1: 1) to afford the desired product (500 mg, 20.5% yield) as yellow solid. LC-MS: m/z 212 (M+H) ⁺ .

[00210] Step D. 2-Chloro-l-(6-(pyrimidin-2-yloxy)-lH-indol-3-yl)ethanone. To a stirred solution of 6-(pyrimidin-2-yloxy)-lH-indole (100 mg, 0.474 mmol) in anhydrous DCM (10 mL) at 0°C was added A1C1 ₃ (189 mg, 1.442 mmol), followed by dropwise addition of 2-chloroacetyl chloride (160 mg, 1.422 mmol) in anhydrous DCM (2 mL). The reaction mixture was stirred at r.t. for 16 hr, then quenched with water and extracted with DCM (20 mL). The organic layer was separated, washed with brine, dried over anhydrous Na ₂ S0 ₄ and concentrated under reduced pressure to give the crude product (160 mg) as yellow solid which was directly used in the next step without any further purification.

LC-MS: m/z 287 (M+H) ⁺ .

[00211] Step E. 2-(5-chloro-lH-benzo[d]imidazol-2-ylthio)-l-(6-(pyrimidin-2- yloxy)-lH -indol- 3-yl)ethanone2. To a solution of 2-chloro-l-(6-(pyrimidin-2-yloxy)-lH-indol-3-yl)ethanone (60 mg, 0.18 mmol) in DMF (1 mL) at r.t. were added 5-chloro-lH-benzo[d]imidazole-2-thiol (60 mg, 0.33 mmol) and K ₂ C0 ₃ (149 mg, 1.08 mmol). The resulting mixture was stirred at r.t. for 1 hr then filtered. The filtrate was purified by prep-HPLC to afford desired product (5.3 mg, 6.8% yield) as yellow solid. LC-MS: m/z 436 (M+H) ⁺ . H NMR (400 MHz, DMSO-d ₆ ) δ 12.84 (s, 1H), 12.18 (s, 1H), 8.63 (d, = 4.8 Hz, 2H),8.58 (s, 1H), 8.13 (d, = 8.4 Hz, 1H), 7.49 (s, 1H), 7.43 (d, = 8.4 Hz, 1H), 7.33 (s, 1H), 7.25 (t, = 4.8 Hz, 1H), 7.13 (d, = 8.8 Hz, 1H), 7.04 (d, = 8.4 Hz, 1H), 4.89 (s, 2H), 4.30 - 4.20 (m, 2H), 1.40 (t, 3H).

[00212] The procedure set forth above was used to produce the following compounds using the appropriate starting materials.

[00213] 2-( 5-Chloro-7-( 2-hydroxypropan-2-yl)- lH-benzo[ d]imidazol-2-ylthio)-l-( 6-(pyrimidin-2- )

[00214] H NMR (400 MHz, DMSO-d ₆ ) δ 12.35-12.14 (m, 2H), 8.63 (d, = 4.8 Hz, 2H), 8.59 (s, 1H), 8.13 (d, = 8.4 Hz, 1H), 7.33-7.00 (m, 5H), 5.46-5.10 (m, 1H), 4.80 (s, 2H), 1.52-1.44 (m, 6H).

[00215] 2-( 5-Chloro-7-( 2-hydroxypropan-2-yl)-lH-benzo[ d]imidazol-2-ylthio)-l-( 6-(pyrazin-2- )

[00216] LC-MS: m/z 494 (M+H) ⁺ . H NMR (400 MHz, OMSO-d ₆ ) δ 13.00 - 12.25 (m, 1H), 12.15 (s, 1H), 8.59 (s, 1H), 8.53 (s, 1H), 8.36 (d, = 2.7 Hz, 1H), 8.19 (dd, = 2.6, 1.3 Hz, 1H), 8.15 (d, = 8.6 Hz, 1H), 7.47 - 6.90 (m, 4H), 5.74 - 4.94 (m, 1H), 4.90 - 4.60 (m, 2H), 1.60 - 1.25 (m, 6H). EXAMPLE 17. PYOCYANIN INHIBITION ASSAY (TOPSEAL AND BREATHABLE SEAL)

[00217] Test compounds were dissolved in DMSO at 10 mM and stored at -20 °C until needed. Test concentrations from 0.0032 to 31.6 micromolar (topseal) or 0.001 micromolar to 31.6 micromolar (breathable seal) were used for each compound. Seven test concentrations were used for each compound in the topseal assay and eight test concentrations were used for each compound in the breathable seal assay.

[00218J P. aeruginosa, strain PA14, was inoculated into 5 mL of LB broth in a 15-mL sterile glass culture tube and incubated overnight at 37°C under shaking (~ 240 rpm). The overnight culture was then diluted in LB Broth to give an OD ₆₀₀ = 0.04 (T ₀ culture). Colony Forming Units (CFU)/mL of the overnight culture and T ₀ culture were determined by a series of 1 : 10 serial dilutions in sterile saline. 4x20 of each dilution was seeded on LB agar plates and incubated at 35±2 °C for 20 hours. CFU/mL was determined from the dilution that gives 2 to 50 colonies/ 20μh.

[00219] For concentration response curve (CRC) experiments, internal standard, 2-((5-nitro-lH- indol-2-yl)thio)-N-(4-phenoxyphenyl)acetamide, a compound previously identified as an MvfR inhibitor (see Example 1 of US 2014-0066454 (WO 2012/116010)), and test compounds were serially diluted in DMSO to 200 times each final concentration (0.5% DMSO final), and ^L/well is dispensed in duplicate into 2 mL wells of a 96-well deep well plate. 200 μυν/εΙΙ of T ₀ culture were added to each internal standard/ test compound well. Wells containing only T ₀ culture (200 200 μυν/εΙΙ) were used to determine pyocyanin control level and wells containing only LB broth (blank wells) were also included in the plate.

[00220] The topseal assay plate was sealed and incubated at 37°C under shaking (-700 rpm) using a microtiter shaker for 24 hours. In the breathable seal assay the plate was sealed with a breathable seal, covered with a lid, and incubated at 37°C under shaking (-700 rpm) using a microtiter shaker for 24 hours. At the end of the incubation period, the plate was centrifuged at 4,000g for 40 minutes at room temperature, 100 μυν/εΙΙ of the supernatant was transferred to a 96-microtiter PS flat-bottom plate, and absorbance at 690 nm was determined using SPECTROstar® Nano microplate reader. Pyocyanin concentration (M) is determined using the equation C = A ₆₉₀ / (ε xd), where ε is the Pyocyanin extinction coefficient, at A ₆₉₀ nm ε is 4,310 M cm and d is the experimentally derived pathlength. The level of pyocyanin in the presence of test compound was expressed as percentage of inhibition with respect to the control. Curve fitting and IC ₅₀ determination were carried out using a four-parameter logistic model using GraphPad Prism v.5.

[00221] Pyocyanin inhibition data obtained in the topseal assay is provided in Table 1. Pyocyanin inhibition data obtained in the breathable assay is provided in Table 2. Three stars (***) indicates an IC ₅₀ < 0.1 μΜ, two stars (**) indicates 0.1 μΜ < IC ₅₀ <1.0 μΜ, one star (*) indicates 1.0 μΜ < IC ₅₀ <10.0 μΜ, no stars beside a compound number indicates that compound has an IC ₅₀ > 10 μΜ. Table 1. Topseal data

Cmp. No. Pyocyanin IC ₅₀ Cmp. No. Pyocyanin IC ₅₀

(μΜ) (μΜ)

1 ** 8 ***

2 ** 9 ***

3 ** 10 ***

4 ** 11 **

5 ** 12 **

6 ** 13 ***

7 ** 20 ***

Table 2. Breathable seal data

Cmp. No. Pyocyanin IC ₅₀ Cmp. No. Pyocyanin IC50

(μΜ) (μΜ)

9 ** 17 **

11 ** 18 **

12 ** 19

13 ** 20 **

14 ** 21

15 ** 22 **

16 ** 23 **

24 **

EXAMPLE 18. PQS INHIBITION ASSAY (24 HOUR INCUBATION, TOPSEAL)

[00222] Test compounds were dissolved in DMSO at 10 mM and stored at -20 °C until needed. Seven test concentrations from 0.0032 micromolar to 31.6 micromolar were used for each compound. P. aeruginosa PA14 wild-type strain was inoculated into 5 mL LB broth in a 15-mLsteril glass culture tube and incubated overnight at 37°C under shaking (~ 240 rpm). The overnight culture was then diluted in LB Broth to give an OD ₆ oo =0.04 (T ₀ culture). Colony Forming Units (CFU)/mL of the overnight culture and T ₀ culture were determined by a series of 1 :10 serial dilutions in sterile saline. 4x20 \L of each dilution is seeded on LB agar plates and incubated at 35±2 °C for 20 hours. CFU/mL was determined from the dilution that gives 2 to 50 colonies/ 20μί.

[00223] For concentration response curve (CRC) experiments, internal standard (2-((5-nitro-lH- indol-2-yl)thio)-N-(4-phenoxyphenyl)acetamide)) and test compounds were serially diluted in DMSO to give 200 times of each final concentration (0.5% DMSO final), and ^L/well was dispensed in duplicate into 2-mL wells of a 96 well deep well plate. 200 μίΛνεΙΙ of T ₀ culture is added to each internal standard/ test compound well used to determine PQS control level. Wells containing only LB broth (blank wells) were also included in the plate.

[00224] The plate is sealed and incubated at 37°C for 24 hours under shaking (-700 rpm) using a microtiter shaker. At the end of the incubation period, 150 μΙ _^ ΛνεΙΙ of the deep well plate culture were transferred in a new non-sterile plate, and 150 μΙ _^ ΛνεΙΙ of methanol containing tetra-deuterated PQS (D4- PQS) and 2% acetic acid were added to each well. The plate was sealed, shaken at high speed for 5 minutes and centrifuged at 4,000g for 40 minutes at room temperature. 100 μΙ _^ ΛνεΙΙ of the supernatant was then transferred to glass vials (vials crimp 0.2 mL) and kept at 4°C until LC-MS/MS analysis.

[00225] PQSs quantification is carried out by analyzing samples in discrete batches together with spiked standards and blank samples.

[00226] Calibration curves were constructed from PQS standards, and respective deuterated PQS is used as an Internal Standards (IS) to calculate the concentration of analytes in the sample and improve the precision of the assay. The back-calculated concentrations of the calibration standards from the calibration curve shall be within ± 20% of the nominal values, the range of the analytical method is determined, and the lower and upper limit of quantification specified.

[00227] Liquid chromatography separations were performed using an Agilent HP 1100 system (Agilent Technologies) coupled with a CTC PAL Autosampler (CTC Analytics AG). Chromatographic retention is obtained using a monolithic column (Chromolith® RP-18, 50 x 4.6 mm). The solvent system consists of water containing 0.1 % (v/v) formic acid (A) and acetonitrile containing 0.1 % (v/v) formic acid (B). The gradient elution profile is chosen as follows: 0 min: 70% A (1500 μί/ητϊη), 0.3 min: 70% A (1500 μί/ητίη), 1.3 min: 5% A (2500 μί/ητίη), 1.6 min: 5% A (2500 μΙ,Δηίη).

[00228] The MS/MS analysis is performed with an API4000 series mass spectrometer (AB Sciex™) operating in Multiple Reaction Monitoring (MRM) mode and equipped with a TIS ion source. The specific ions monitored were PQS (m z 260 -> m z 175) and D4-PQS (m z 264 -> m z 179).

The computer systems used in this study to acquire and quantify data may be an Analyst™ vl.5.

[00229] PQS in the presence of different concentrations of test compound were expressed as percentage of inhibition of the basal level in control samples. Curve fitting and IC ₅₀ estimations were carried out using a four parameter logistic model using GraphPad Prism v.5 program.

[00230] PQS 24 hour inhibition data is provided in Table 3. Typically compounds that exhibited an IC ₅₀ of less than 10 micromolar in the Pyocyanin inhibition assay were tested in the PQS 24 hour inhibition assay. Three stars (***) indicates an IC ₅₀ < 1.0 μΜ, two stars (**) indicates 1.0 μΜ < IC ₅₀ <5.0 μΜ, one star (*) indicates 5.0 μΜ < IC ₅₀ <10.0 μΜ, no stars beside a compound number indicates that compound has an IC ₅₀ > 10 μΜ. Table 3

Cmp. No. PQS (24 hr)

IC ₅₀ (μΜ)

4

5 ***

6 ***

7 ***

8 ***

9 ***

EXAMPLE 19. ASSAY FOR PQS AND HHQ INHIBITION (6 HOUR INCUBATION)

[00231] A concentration range with 7 test concentrations was chosen for each test compound. Typically concentration ranges for the PQS and HHQ assays were from 0.03 to 31.6 μΜ. Compounds were prepared in solvent, usually DMSO, at either 200 or 500 times the final test concentration.

[00232] P. aeruginosa PA 14 wild-type strain was inoculated into 5 ml LB broth in a 15 ml sterile glass culture tube and incubated overnight at 37°C under shaking (~ 240 rpm). After overnight incubation, the bacterial culture was diluted in fresh LB broth to an OD600nm=0.04, and 5 mL aliquots of the diluted bacterial culture were distributed into 15-mL glass tubes. 10 of test compound at 500 times the final test concentration or 25μί of test compound at 200 times the final test concentration or solvent alone was added to each of the bacterial suspension tubes, and the tubes were incubated at 37°C under shaking (300rpm) for 6 hours. At the end of the incubation period, a 0.5 mL aliquot of bacterial suspension was transferred from each 15 mL glass tube, after shaking, to a 2 mL polypropylene vial. 0.5 mL of methanol-containing D4-PQS (deuterated-PQS), D4-HHQ (deuterated-HHQ) and 2% acetic acid were added to each 2 mL vial and each vial was vigorously shaken. The vials were then centrifuged for 5 minutes at 12,000g, and 200 μί of supernatant from each vial is transferred to a glass vial (vials crimp 0.2mL), and the sealed vials were kept at 4°C until LC-MS/MS analysis was performed. HAQs quantification is carried out analyzing samples in discrete batches together with spiked standards and blank samples. Calibration curves were constructed from HAQs standards, and respective deuterated forms were used as Internal Standards (IS), to calculate the concentration of analytes in the sample and improve the precision of the assay. The back-calculated concentrations of the calibration standards from the calibration curve are within ± 20% of the nominal values, the range of the analytical method was determined, and the lower and upper limit of quantification specified.

[00233] Liquid chromatography separations were performed using Agilent HP1100 system (Agilent Technologies) coupled with a CTC PAL Autos ampler (CTC Analytics AG). Chromatographic retention is obtained using a monolithic column (Chromolith® RP-18, 50 x 4.6 mm). The solvent system consists of water containing 0.1 % (v/v) formic acid (A) and acetonitrile containing 0.1 % (v/v) formic acid (B). The gradient elution profile wass chosen as follows: 0 min: 70% A (1500 μυηή), 0.3 min: 70% A (1500 μΙ7ητίη), 1.3 min: 5% A (2500 μϋιηιη), 1.6 min: 5% A (2500 μΙ7ητίη).

[00234] The MS/MS analysis was performed with a API4000 series mass spectrometer (AB Sciex™) operating in Multiple Reaction Monitoring (MRM) mode and equipped with a TIS ion source. The specific ions monitored were PQS (m/z 260 -> m/z 175), D4-PQS (m/z 264 -> m/z 179), HHQ (m/z 244 -> m/z 159) and D4-HHQ (m/z 248 -> m/z 163). PQS and HHQ in presence of different test compound concentrations were expressed as percentage of inhibition of the basal level in control samples.

[00235] PQS 6 hour inhibition data is provided in Table 4. HHQ 6 hour inhibition data is provided in Table 5. Typically compounds that exhibited an IC ₅₀ of less than 1 micromolar in the Pyocyanin inhibition assay were tested in the PQS and HHQ 6 hour inhibition assays. Three stars (***) indicates an IC ₅₀ < 1.0 μΜ, two stars (**) indicates 1.0 μΜ < IC ₅₀ <5.0 μΜ, one star (*) indicates 5.0 μΜ < IC ₅₀ < 10.0 μΜ, no stars beside a compound number indicates that compound has an IC ₅₀ > 10 μΜ.