Haloindole macrocyclic compounds for the treatment of cancer

The present invention relates to organic compounds useful for therapy and/or prophylaxis in a mammal, and in particular to inhibition of KRAS G12C useful for treating cancers.

FIELD OF THE INVENTION

RAS is one of the most well-known proto-oncogenes. Approximately 30% of human cancers contain mutations in three most notable members, KRAS, HRAS, and NRAS, making them the most prevalent oncogenic drivers. KRAS mutations are generally associated with poor prognosis especially in colorectal cancer, pancreatic cancer, lung cancers. As the most frequently mutated RAS isoform, KRAS has been intensively studied in the past years. Among the most commonly occurring KRAS alleles (including G12D, G12V, G12C, G13D, G12R, G12A, G12S, Q61H, etc), G12C, G12D, G12V represent more than half of all K-RAS-driven cancers across colorectal cancer (CRC), pancreatic ductal adenocarcinoma (PDAC), lung adenocarcinoma (LUAD). Of note, KRAS wild-type amplifications are also found in around 7% of all KRAS- altered cancers (ovarian, esophagogastric, uterine), ranking among the top alterations.

All RAS proteins belong to a protein family of small GTPases that hydrolyze GTP to GDP. KRAS is structurally divided into an effector binding lobe followed by the allosteric lobe and a carbo xy-terminal region that is responsible for membrane anchoring. The effector lobe comprises the P-loop, switch I, and switch II regions. The switch I/II loops play a critical role in KRAS downstream signaling through mediating protein-protein interactions with effector proteins that include RAF in the mitogen-activated protein kinase (MAPK) pathway or PI3K in the phosphatidylinositol 3 -kinase (PI3K)/protein kinase B (AKT) pathway.

KRAS protein switches between an inactive to an active form via binding to GTP and GDP, respectively. Under physiological conditions, the transition between these two states is regulated by guanine nucleotide exchange factors (GEFs), such as Son Of Sevenless Homolog 1 (S0S1), or GTPase- activating proteins (GAPs) that involve catalyzing the exchange of GDP for GTP, potentiating intrinsic GTPase activity or accelerating RAS -mediated GTP hydrolysis. In response to extracellular stimuli, the inactive RAS -GDP is converted to active RAS -GTP which directly binds to RAF RAS binding domains (RAF ^RBD ), recruiting RAF kinase family from cytoplasm to membranes, where they dimerize and become active. The activated RAF subsequently carries out a chain of phosphorylation reactions to its downstream Mitogen- activated protein kinase (MEK) and extracellular signal-regulated kinase (ERK), and propagates the growth signal. Of the RAF family of protein kinases (three known isoforms ARAF, BRAF, CRAF/RAF1), BRAF is most frequently mutated and remains the most potent activator of MEK. Despite that individual RAS and RAF family members revealed distinct binding preferences, all RAFs possess the conserved RBD for forward transmission of MAPK singnaling, frequently used for characterize KRAS inhibition (e.g. KRAS-BRAF ^RBD herein). For KRAS, mutations at positions 12, 13, 61, and 146 lead to a shift toward the active KRAS form through impairing nucleotide hydrolysis or activating nucleotide exchange, leading to hyper-activation of the MAPK pathway that results in tumorigenesis.

Despite its well-recognized importance in cancer malignancy, continuous efforts in the past failed to develop approved therapies for KRAS mutant cancer until recently, the first selective drug AMG510 has fast approval as second line treatment in KRAS G12C driven non-small cell lung cancer (NSCEC). Nevertheless, the clinical acquired resistance to KRAS G12C inhibitors emerge rigorously with disease progresses after around 6 month of treatment. All of the mutations converge to reactivate RAS-MAPK signaling, with secondary RAS mutants at oncogenic hotspots (e.g. G12/G13/Q61) and within the switch II pocket (e.g. H95, R68, and Y96) have been observed; moreover, over 85% of all KRAS-mutated or wild-type amplified driven cancers still lack novel agents. Altogether, both the myriad of escape mechanism and various oncogenic alleles, highlight the urgent medical need for additional KRAS therapies. As such, we invented oral compounds that target and inhibit KRAS alleles for the treatment of KRAS mutant driven cancers.

SUMMARY OF THE INVENTION

The present invention relates to novel compounds of formula (I), wherein 1 R is , 1-oxo-2,7-diazaspiro[4.4]nonanyl substituted by (dihaloC _1-6 alkyl)carbonyl, 1-oxo-2,8-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl, 1-oxo-2,9-diazaspiro[5.5]undecanyl substituted by (dihaloC1-6alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.4]octanyl substituted by (dihaloC _1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonanyl substituted by (dihaloC1-6alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl or a ^{zetidinyloxyC} 1-6 ^{alkyl substituted by (dihaloC} 1-6 ^{alkyl)carbonyl;} wherein R ⁸ is C _1-6 alkyl; R ⁹ is piperidinyl twice substituted by halogen and (dihaloC1-6alkyl)carbonyl, or pyrrolidinyl substituted by (dihaloC _1-6 alkyl)carbonyl; R ² is C _1-6 alkyl; R ³ is H, C1-6alkyl or halogen; R ⁴ is H or halogen; R ⁵ is C _1-6 alkyl or haloC _1-6 alkyl; R ⁶ is C _1-6 alkoxyC _1-6 alkyl; R ⁷ is morpholinyl, (haloC1-6alkyl)piperazinyl or C1-6alkylpiperazinyl; A ¹ is thiazolylene or phenylene substituted by hydroxy; A ² is C _1-6 alkylene; with the proviso that R ³ and R ⁴ are not H simultaneously; or a pharmaceutically acceptable salt thereof. The invention also relates to their manufacture, medicaments based on a compound in accordance with the invention and their production as well as the use of compounds of formula (I) or (Ia) thereof as inhibitor of KRAS. The compounds of formula (I) or (Ia) show good KRAS inhibition for G12C, G12D and G12V. In another embodiment, the compounds of this invention showed superior cancer cell inhibition and human hepatocyte stability. In addition, the compounds of formula (I) or (Ia) also show good or improved cytotoxicity, solubility profiles. Furthermore, the compound of current invention addressed GSH toxicity issue comparing with the reference compounds. BRIEF DESCRIPTION OF THE FIGURE Figure 1. X-ray crystallographic analysis of compound 4f 1. Figure 2. X-ray crystallographic analysis of compound 4f 2. Figure 3. X-ray crystallographic analysis of compound M2. DETAILED DESCRIPTION OF THE INVENTION DEFINITIONS The term “C1-6alkyl” denotes a saturated, linear or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms, for example methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl and the like. Particular “C _1-6 alkyl” groups are methyl, ethyl and n-propyl. The term “C1-6alkylene” denotes a linear or branched saturated divalent hydrocarbon group of 1 to 6 carbon atoms or a divalent branched saturated divalent hydrocarbon group of 3 to 6 carbon atoms. Examples of C _1-6 alkylene groups include methylene, ethylene, propylene, 2- methylpropylene, butylene, 2-ethylbutylene, pentylene, hexylene. The term “halogen” and “halo” are used interchangeably herein and denote fluoro, chloro, bromo, or iodo. The term “haloindole” denotes an indole wherein one or more of the hydrogen atoms have been replaced by same or different halogen atoms. T ^{he term “dihaloC} 1-6 ^{alkyl” denotes a C} 1-6 ^{alkyl group wherein two of the hydrogen atoms} of the C _1-6 alkyl group have been replaced by same or different halogen atoms. Examples of dihaloC1-6alkyl include, difluoro- or chloro(fluoro)-methyl, -ethyl or -propyl, particularly difluoropropyl, difluoromethyl, difluoroethyl or chloro(fluoro)methyl. The term “haloC _1-6 alkyl” denotes a C _1-6 alkyl group wherein at least one of the hydrogen atoms of the C1-6alkyl group have been replaced by same or different halogen atoms. Examples of haloC1-6alkyl include fluoro, difluoro- or chloro(fluoro)-methyl, -ethyl or -propyl, for example fluoromethyl, difluoropropyl, difluoromethyl, difluoroethyl, chloro(fluoro)methyl, trifluoroethyl, or trifluoromethyl. The term “phenylene” denotes a divalent phenyl group. The term “piperidinylene” denotes a divalent piperidinyl group. The term “pyrrolidinylene” denotes a divalent pyrrolidinyl group. The term “thiazolylene” denotes a divalent thiazolyl group. The term “oxo” denotes a divalent oxygen atom =O. The term “azetidinyloxy” denotes azetidinyl-O-.

The term “dimethylmethylene” denotes

The term “protecting group” denotes the group which selectively blocks a reactive site in a multifunctional compound such that a chemical reaction can be carried out selectively at another unprotected reactive site in the meaning conventionally associated with it in synthetic chemistry.

Protecting groups can be removed at the appropriate point. Exemplary protecting groups are amino -protecting groups, carboxy-protecting groups or hydroxy-protecting groups.

The skilled of the art would understand that the following structures of compounds of formula (la) and (la’) are equal especially for the chiral centers:

The term “pharmaceutically acceptable salts” denotes salts which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid and base addition salts.

The term “pharmaceutically acceptable acid addition salt” denotes those pharmaceutically acceptable salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-tolucncsulfonic acid, and salicyclic acid.

The term “pharmaceutically acceptable base addition salt” denotes those pharmaceutically acceptable salts formed with an organic or inorganic base. Examples of acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Salts derived from pharmaceutically acceptable organic nontoxic bases includes salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperizine, piperidine, A-ethylpiperidine, and polyamine resins.

The term “A pharmaceutically active metabolite” denotes a pharmacologically active product produced through metabolism in the body of a specified compound or salt thereof. After entry into the body, most drugs are substrates for chemical reactions that may change their physical properties and biologic effects. These metabolic conversions, which usually affect the polarity of the compounds of the invention, alter the way in which drugs are distributed in and excreted from the body. However, in some cases, metabolism of a drug is required for therapeutic effect.

The term “therapeutically effective amount” denotes an amount of a compound or molecule of the present invention that, when administered to a subject, (i) treats or prevents the particular disease, condition or disorder, (ii) attenuates, ameliorates or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition or disorder described herein. The therapeutically effective amount will vary depending on the compound, the disease state being treated, the severity of the disease treated, the age and relative health of the subject, the route and form of administration, the judgement of the attending medical or veterinary practitioner, and other factors.

The term “pharmaceutical composition” denotes a mixture or solution comprising a therapeutically effective amount of an active pharmaceutical ingredient together with pharmaceutically acceptable excipients to be administered to a mammal, e.g., a human in need thereof.

The terms “pharmaceutically acceptable excipient”, “pharmaceutically acceptable carrier” and “therapeutically inert excipient” can be used interchangeably and denote any pharmaceutically acceptable ingredient in a pharmaceutical composition having no therapeutic activity and being non-toxic to the subject administered, such as disintegrators, binders, fillers, solvents, buffers, tonicity agents, stabilizers, antioxidants, surfactants, carriers, diluents or lubricants used in formulating pharmaceutical products. INHIBITOR OF KRAS The present invention relates to (i) a compound of formula (I), wherein R ¹ is , 1-oxo-2,7-diazaspiro[4.4]nonanyl substituted by (dihaloC1-6alkyl)carbonyl, 1-oxo-2,8-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl, 1-oxo-2,9-diazaspiro[5.5]undecanyl substituted by (dihaloC _1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.4]octanyl substituted by (dihaloC _1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonanyl substituted by (dihaloC1-6alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl or azetidinyloxyC _1-6 alkyl substituted by (dihaloC _1-6 alkyl)carbonyl; wherein R ⁸ is C1-6alkyl; R ⁹ is piperidinyl twice substituted by halogen and (dihaloC1-6alkyl)carbonyl, or pyrrolidinyl substituted by (dihaloC _1-6 alkyl)carbonyl; R ² is C _1-6 alkyl; R ³ is H, C1-6alkyl or halogen; R ⁴ is H or halogen; R ⁵ is C1-6alkyl or haloC1-6alkyl; R ⁶ is C _1-6 alkoxyC _1-6 alkyl; R ⁷ is morpholinyl, (haloC1-6alkyl)piperazinyl or C1-6alkylpiperazinyl; A ¹ is thiazolylene or phenylene substituted by hydroxy; A ² is C _1-6 alkylene; with the proviso that R ³ and R ⁴ are not H simultaneously; or a pharmaceutically acceptable salt thereof. Another embodiment of present invention is (ii) a compound of formula (Ia), wherein R ¹ is , 1-oxo-2,7-diazaspiro[4.4]nonanyl substituted by (dihaloC _1-6 alkyl)carbonyl, 1-oxo-2,8-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl, 1-oxo-2,9-diazaspiro[5.5]undecanyl substituted by (dihaloC1-6alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.4]octanyl substituted by (dihaloC _1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonanyl substituted by (dihaloC1-6alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl or azetidinyloxyC _1-6 alkyl substituted by (dihaloC _1-6 alkyl)carbonyl; wherein R ⁸ is C _1-6 alkyl; R ⁹ is piperidinyl twice substituted by halogen and (dihaloC1-6alkyl)carbonyl, or pyrrolidinyl substituted by (dihaloC _1-6 alkyl)carbonyl; R ² is C1-6alkyl; R ³ is H, C _1-6 alkyl or halogen; R ⁴ is H or halogen; R ⁵ is C _1-6 alkyl or haloC _1-6 alkyl; R ⁶ is C _1-6 alkoxyC _1-6 alkyl; R ⁷ is morpholinyl, (haloC1-6alkyl)piperazinyl or C1-6alkylpiperazinyl; A ¹ is thiazolylene or phenylene substituted by hydroxy; A ² is C _1-6 alkylene; with the proviso that R ³ and R ⁴ are not H simultaneously; or a pharmaceutically acceptable salt thereof. A further embodiment of present invention is (iii) a compound of formula (I) or (Ia) according to (i) or (ii), or a pharmaceutically acceptable salt thereof, wherein R ¹ is , 1-oxo-2,7-diazaspiro[4.4]nonanyl substituted by (dihaloC1-6alkyl)carbonyl, 1-oxo-2,8-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl, 1-oxo-2,9-diazaspiro[5.5]undecanyl substituted by (dihaloC _1-6 alkyl)carbonyl, or 5-oxo-2,6-diazaspiro[3.4]octanyl substituted by (dihaloC _1-6 alkyl)carbonyl; wherein R ⁸ is C1-6alkyl; R ⁹ is piperidinyl twice substituted by halogen and (dihaloC _1-6 alkyl)carbonyl, or pyrrolidinyl substituted by (dihaloC _1-6 alkyl)carbonyl. A further embodiment of present invention is (iv) a compound of formula (I) or (Ia), according to any one of (i) to (iii), or a pharmaceutically acceptable salt thereof, wherein R ¹ is , 1-oxo-2,7-diazaspiro[4.4]nonan-2-yl substituted by chloro(fluoro)acetyl, 1-oxo-2,8-diazaspiro[4.5]decan-2-yl substituted by chloro(fluoro)acetyl, 1-oxo-2,9-diazaspiro[5.5]undecan-2-yl substituted by chloro(fluoro)acetyl, or 5-oxo-2,6-diazaspiro[3.4]octan-6-yl substituted by chloro(fluoro)acetyl; wherein R ⁸ is methyl;

R ⁹ is piperidinyl twice substituted by fluoro and chloro (fluoro) acetyl, or pyrrolidinyl substituted by chloro(fluoro)acetyl.

A further embodiment of present invention is (v) a compound of formula (I) or (la) according to any one of (i) to (iv), wherein R ¹ is (5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo- 2,7-diazaspiro[4.4]nonan-2-yl, (5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl, 8-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diazaspiro[4.5]d ecan-2- yl, 9-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diazaspiro[5.5]u ndecan-2-yl, 2-[(2R)-2-chloro-2- fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octan-6-yl, l-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-4- piperidinyl or l-[(2R)-2-chloro-2-fluoro-acetyl]pyrrolidin-3-yl.

A further embodiment of present invention is (vi) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (v), wherein R ² is isopropyl.

A further embodiment of present invention is (vii) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (vi), wherein R ³ is H, methyl or fluoro.

A further embodiment of present invention is (viii) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (vii), wherein R ³ is fluoro.

A further embodiment of present invention is (ix) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (viii), wherein R ⁴ is H or fluoro.

A further embodiment of present invention is (x) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (ix), wherein R ⁴ is H.

A further embodiment of present invention is (xi) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (x), wherein R ⁵ is ethyl or 2,2,2-trifluoroethyl.

A further embodiment of present invention is (xii) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xi), wherein 1- methoxyethyl.

A further embodiment of present invention is (xiii) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xii), wherein R ⁷ is morpholinyl, 4-(2,2,2-trifluoroethyl)piperazin-l-yl or 4-methylpiperazin-l-yl. A further embodiment of present invention is (xiv) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xiii), wherein A ¹ is

, wherein bond “a” connects to indole ring.

A further embodiment of present invention is (xv) a compound of formula (I) or (la), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xiv), wherein A ² is dimethylmethylene .

Another embodiment of present invention is (xvi) a compound of formula (I) or (la), , according to (i) or (ii), wherein

R ¹ is l-oxo-2,7-diazaspiro[4.4]nonanyl substituted by (dihaloC1-6alkyl)carbonyl, l-oxo-2,8-diazaspiro[4.5]decanyl substituted by (dihaloC1-6alkyl)carbonyl, l-oxo-2,9-diazaspiro[5.5]undecanyl substituted by (dihaloC1-6alky l)carbonyl, or 5-oxo-2,6-diazaspiro[3.4]octanyl substituted by (dihaloCi-ealkyl)carbonyl; wherein R ⁸ is C1-6alkyl;

R ⁹ is piperidinyl twice substituted by halogen and (dihaloC1-6alky l)carbonyl, or pyrrolidinyl substituted by (dihaloC1-6alky l)carbonyl;

R ² is Ci-ealkyl;

R ³ is halogen;

R ⁴ is H;

R ⁵ is Ci-ealkyl or haloCi-ealkyl;

R ⁶ is Ci-ealkoxyCi-ealkyl;

R ⁷ is morpholinyl, (haloCi-ealkyl)piperazinyl or Ci-ealkylpiperazinyl;

A ¹ is , wherein bond “a” connects to indole ring;

A ² is C1-6alkylene; or a pharmaceutically acceptable salt thereof. Another embodiment of present invention is (xvii) a compound of formula (I) or (la), , according to (xvi), wherein

R ¹ is (5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7-diazaspiro[ 4.4]nonan-2-yl, (5S)-7- [(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7-diazaspiro[4.4]non an-2-yl, 8-[(2R)-2-chloro- 2-fluoro-acetyl]-l-oxo-2,8-diazaspiro[4.5]decan-2-yl, 9-[(2R)-2-chloro-2-fluoro-acetyl]- l-oxo-2,9-diazaspiro[5.5]undecan-2-yl, 2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6- diazaspiro[3.4]octan-6-yl, l-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-4-piperidinyl or 1- [(2R)-2-chloro-2-fluoro-acetyl]pyrrolidin-3-yl;

R ² is isopropyl;

R ³ is fluoro;

R ⁴ is H;

R ⁵ is ethyl or 2,2,2-trifluoroethyl;

R ⁶ is (IS)-l-methoxyethyl;

R ⁷ is morpholinyl, 4-(2,2,2-trifluoroethyl)piperazin-l-yl or 4-methylpiperazin-l-yl;

A ¹ is , wherein bond “a” connects to indole ring;

A ² is dimethylmethylene; or a pharmaceutically acceptable salt thereof.

Another embodiment of present invention is (xviii) a compound of formula (I) or (la) selected from the following:

(35)-l-[(2R)-2-chloro-2-fluoro-acetyl]-A-[(15)-l-[[(75,13 5)-25-fluoro-(20Af)-20-[2-[(15)- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-

25 9 13 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-A-methyl-pyrrolidine-3- carboxamide;

(35)-l-[(2R)-2-chloro-2-fluoro-acetyl]-A-[(15)-l-[[(75,13 5)-24-fluoro-(20Af)-20-[2-[(15)- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-A-methyl-pyrrolidine-3- carboxamide;

(35)-l-[(2R)-2-chloro-2-fluoro-acetyl]-A-[(15)-l-[[(75,13 5) -(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17,24 -trimethyl-8,14-dioxo-21-(2,2,2- 25 9 13 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamo yl]-2-methyl-propyl]-Af-methyl-pyrrolidine-3- carboxamide;

(2S)-2-[(57?)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2, 7-diazaspiro[4.4]nonan-2-yl]-/V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(5S)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-/V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[9-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diaz aspiro[5.5]undecan-2-yl]-/V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(35)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(15)-l-[[(85,1 45)-25-fluoro-4-hydroxy-(21Af)-

21-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3- pyridyl]-18,18-dimethyl-9,15-dioxo-

2 10 14 25 27

22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacycl o[18.5.2.1 .1 ’ .0 ]nonacosa- l(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methy l-propyl]-JV-methyl-pyrrolidine- 3 -carboxamide; l-[(2/?)-2-chloro-2-fluoro-acetyl]-4-fluoro-^-[(15)-l-[[(75, 135) -(20Af)-20-[2-[(15)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17,24 -trimethyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-/V-methyl-piperid ine-4- carboxamide; l-[(2/?)-2-chloro-2-fluoro-acetyl]-4-fluoro-^-[(15)-l-[[(75, 135)-24-fluoro-(20Af)-20-[2- [(15)- l-methoxyethyl]-5-(4-methylpiperazin- 1 -yl) - 3 -pyridyl] - 17, 17-dimethyl-8, 14-dioxo-21-

25 9 13 2226 (2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapenta cyclo[17.5.2.1 ’ .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamo yl]-2-methyl-propyl]-Af-methyl-piperidine-4- carboxamide;

(2S)-2-[2-[(27?)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.5]nonan-6-yl]-2V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(27?)-2-[[l-[(27?)-2-chloro-2-fluoro-acetyl]azetidin-3-yl ]oxymethyl]-A^-[(7S,135')-24-fluoro- (20A7)-20-[2-[(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl )-3-pyridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tet razapentacyclo-

[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-me thyl-butanamide;

(25 ^, )-2-[[l-[(27?)-2-chloro-2-fluoro-acetyl]azetidin-3-yl] oxymethyl]-A^-[(7S,135')-24-fluoro- (20A7)-20-[2-[(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl )-3-pyridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[8-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diaz aspiro[4.5]decan-2-yl]-/V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(5S)-2-[(27?)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7 -diazaspiro[4.5]decan-7-yl]-A^- [(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(57?)-2-[(27?)-2-chloro-2-fluoro-acetyl]-6-oxo-2, 7-diazaspiro[4.5]decan-7-yl]-/V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[9-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diaz aspiro[5.5]undecan-2-yl]-A^- [(7S,13S)-25-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl- butanamide ;

(2S)-2-[(57?)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7 -diazaspiro[4.5]decan-7-yl]-/V- [(7S, 13S)-21 -cthyl-24-fluoro-(20A7)-20-[2-[( l S)- l -incthoxycthyl]-5-(4-mcthylpipcrazin- l -yl)-3- pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl- butanamide ;

(25 ^, )-2-[(5S)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7- diazaspiro[4.5]decan-7-yl]-A^- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethy l]-5-(4-methylpiperazin-l-yl)-3- pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl- butanamide ;

(35)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(15)-l-[[(75,1 35)-24-fluoro-(20Af)-20-[2-[(15)- l-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-

2 5 9 13 222 26

15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamo yl]-2-methyl-propyl]-N-methyl-pyrrolidine-3- carboxamide;

(2S)-2-[2-[(27?)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-A^- [(7S,13S)-24-fluoro-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylp iperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl- butanamide ;

(35)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(15)-l-[[(75,1 35)-24-fluoro-(20Af)-20-[2-[(15)- l-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-l-yl]-3 -pyridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-Af-methyl-pyrrolidine-3-carbo xamide;

(3S)-l-[(27?)-2-chloro-2-fluoro-acetyl]-AH(159-l-[[(8S,14 S)-26-fluoro-4-hydroxy-(21A7)-

21-[2-[(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3- pyridyl]-18,18-dimethyl-9,15-dioxo-

2 10 14 25 27

22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacycl o[18.5.2.1 .1 ’ .0 ]nonacosa- l(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methy l-propyl]-A^-methyl-pyrrolidine- 3 -carboxamide;

(2S)-2-[8-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diaz aspiro[4.5]decan-2-yl]-iV- [(7S,13S)-21-ethyl-24-fluoro-(20A7)-20-[2-[(lS)-l-methoxyeth yl]-5-(4-methylpiperazin-l-yl)-3- pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[2-[(27?)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-A^- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethy l]-5-(4-methylpiperazin-l-yl)-3- pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(57?)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2, 7-diazaspiro[4.4]nonan-2-yl]-iV- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethy l]-5-(4-methylpiperazin-l-yl)-3- pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(5S)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-iV- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethy l]-5-(4-methylpiperazin-l-yl)-3- pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 , 22(26), 23-hexaen-7 -yl]-3- methyl-butanamide ;

(35)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(15)-l-[[(75,1 35)-21-ethyl-24-fluoro-(20Af)-20- [2-[( IS)- l-methoxyethyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-

2 5 9 13 222 2^ oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-/V-methyl-pyrroli dine-3- carboxamide;

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,1 3S)-21-ethyl-24-fluoro-(20Af)-20- [2-[(lS)-l-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazi n-l-yl]-3-pyridyl]-17,17-dimethyl- 2 5 9 13 22 2& 8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2 .1 ’ .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-/V-methyl-pyrroli dine-3- carboxamide; l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,13S)-21-e thyl-24-fluoro-(20Af)-20-[2-

[( IS)- l-methoxyethyl]-5-(4-methylpiperazin- 1 -yl) - 3 -pyridyl] - 17, 17-dimethyl-8, 14-dioxo- 15-

25 9 13 22226 oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-/V-methy l- piperidine-4-carboxamide;

(2S)-2-[(57?)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2, 7-diazaspiro[4.4]nonan-2-yl]-/V- [(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyeth yl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(5S)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-/V- [(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyeth yl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[9-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diaz aspiro[5.5]undecan-2-yl]-/V- [(7S,13S)-21-ethyl-24-fluoro-(20A7)-20-[2-[(lS)-l-methoxyeth yl]-5-(4-methylpiperazin-l-yl)-3- pyridyl]- 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[2-[(27?)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-/V- [(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyeth yl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[8-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diaz aspiro[4.5]decan-2-yl]-iV- [(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyeth yl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(27?)-2-[[l-[(27?)-2-chloro-2-fluoro-acetyl]azetidin-3-yl ]oxymethyl]-A^-[(7S,13S)-21-ethyl- 24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiper azin-l-yl)-3-pyridyl]-17,17- 25 9 13 22226 dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyc lo[17.5.2.1 ’ .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-butanamide;

(2S)-2-[[l-[(2R)-2-chloro-2-fluoro-acetyl]azetidin-3-yl]o xymethyl]-A^-[(75',135')-21-ethyl- 24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiper azin-l-yl)-3-pyridyl]-17,17- 25 9 13 2226 dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyc lo[17.5.2.1 ’ .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-butanamide;

(2S)-2-[9-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diaz aspiro[5.5]undecan-2-yl]-/V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-[4-( 2,2,2-trifluoroethyl)piperazin-l- yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoro ethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-N- [(7S,13S)-24-fluoro-20-[2-[(lS)-l-methoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-/V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-[4-( 2,2,2-trifluoroethyl)piperazin-l- yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoro ethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ; l-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-^-[(15)-l-[[(75,1 35)-24-fluoro-(20Af)-20-[2- [(1S)- l-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin- l-yl]-3-pyridyl]- 17, 17-dimethyl-8, 14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamoyl]-2-mcthyl-propyl]-A ^/ -mcthyl-pipcridinc-4-carboxainidc;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaza spiro[3.4]octan-6-yl]-2V- [(7S,13S)-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-[4-( 2,2,2-trifluoroethyl)piperazin-l- yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoro ethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ; (2S)-2-[8-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diazasp iro[4.5]decan-2-yl]-iV- [(7S,13S)-24-fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl]-5-[4- (2,2,2-trifluoroethyl)piperazin-l- yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoro ethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[9-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diaz aspiro[5.5]undecan-2-yl]-/V- [(75,135)-21-ethyl-24-fluoro-(20Af)-20-[2-[(15)-l-methoxyeth yl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ; l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(15)-l-[[(75,135)-21-e thyl-24-fluoro-(20Af)-20-[2- [(15)-l-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-l -yl]-3-pyridyl]-17,17-dimethyl-8,14- 25 9 13 22226 dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-/V-methy l- piperidine-4-carboxamide;

(2S)-2-[8-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diaz aspiro[4.5]decan-2-yl]-/V- [(75, 13S)-21 -ethyl-24-fluoro-(20A7)-20- [2- [( 15)- 1 -methoxyethyl] -5-morpho lino-3 -pyridyl] -

17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9,21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(5S)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-iV- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethy l]-5-morpholino-3-pyridyl]-

17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[(57?)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2, 7-diazaspiro[4.4]nonan-2-yl]-iV- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethy l]-5-morpholino-3-pyridyl]-

17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ;

(2S)-2-[2-[(27?)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-A^- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(15)-l-methoxyethy l]-5-morpholino-3-pyridyl]-

17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia-9, 21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3- methyl-butanamide ; l-[(2/?)-2-chloro-2-fluoro-acetyl]-A-[(lS)-l-[[(7S,13S)-21-e thyl-24-fluoro-(20Af)-20-[2- [( IS)- l-methoxyethyl]-5-morpholino-3-pyridyl] - 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen- 7-yl]carbamoyl]-2-methyl-propyl]-4-fhioro-A-methyl-piperidin e-4-carboxamide;

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-A-[(lS)-l-[[(7S,1 3S)-21-ethyl-24-fluoro-(20Af)-20- [ 2- [ ( 1 S)- l-methoxyethyl]-5-morpholino-3-pyridyl] - 17, 17-dimethyl-8, 14-dioxo- 15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen- 7-yl]carbamoyl]-2-methyl-propyl]-A-methyl-pyrrolidine-3-carb oxamide;

(2S)-2-[(5S)-7-[(27?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-A- [(7S,13S)-24-fhioro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-morp holino-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ ,0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide; and

(2S)-2-[2-[(27?)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-A- [(7S,13S)-24-fhioro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-morp holino-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ ,0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide ; or a pharmaceutically acceptable salt thereof.

Another embodiment of present invention is related to (xix) a compound which has the following structure:

(RM44-A).

Another embodiment of present invention is related to (xx) a process for the preparation of a compound according to any one of (i) to (xix) comprising any of the following steps: a) coupling reaction between compound of formula (II), in the presence of a coupling reagent and a base to form the compound of formula (I); b) coupling reaction between compound of formula (VI),

and acid (VII), (VII), in the presence of a coupling reagent and a base to form the compound of formula (VIII),

5 c) coupling reaction between compound of formula (XI),

and acid (XII), (XII), in the presence of a coupling reagent and a base to form the compound of formula (XIII), wherein Q is unsubstituted or substituted piperidinylene or pyrrolidinylene; T is dihaloCi- ealkyl; R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , A ¹ and A ² are defined as in any one of (i) to (xvii); the coupling reagent in step a) to c) is T3P, HATU, PyBOP or EDCI/HOBt; the base in step a) to c) is TEA, DIEPA or DMAP.

Another embodiment of present invention is (xxi) a compound or pharmaceutically acceptable salt according to any one of (i) to (xix) for use as therapeutically active substance. Another embodiment of present invention is (xxii) a pharmaceutical composition comprising a compound in accordance with any one of (i) to (xix) and a pharmaceutically acceptable excipient. Another embodiment of present invention is (xxiii) the use of a compound according to any one of (i) to (xix) for treating a KRAS G12C protein-related disease.

Another embodiment of present invention is (xxiv) the use of a compound according to any one of (i) to (xix) for treating a KRAS G12C, G12D and G12V protein-related disease.

Another embodiment of present invention is (xxv) the use of a compound according to any one of (i) to (xix) for inhibiting RAS interaction with downstream effectors, wherein the downstream effectors are RAF and PI3K.

Another embodiment of present invention is (xxvi) the use of a compound according to any one of (i) to (xix) for inhibiting the propagating oncogenic MAPK and PI3K signaling.

Another embodiment of present invention is (xxvii) the use of a compound according to any one of (i) to (xix) for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma ovarian cancer and endometrial cancer.

Another embodiment of present invention is (xxviii) the use of a compound according to any one of (i) to (xix) for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

Another embodiment of present invention is (xxix) a compound or pharmaceutically acceptable salt according to any one of (i) to (xix) for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

Another embodiment of present invention is (xxx) the use of a compound according to any one of (i) to (xix) for the preparation of a medicament for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

Another embodiment of present invention is (xxxi) a method for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer, which method comprises administering a therapeutically effective amount of a compound as defined in any one of (i) to

(xix).

Another embodiment of present invention is (xxxii) a compound or pharmaceutically acceptable salt according to any one of (i) to (xix), when manufactured according to a process of

(xx). PHARMACEUTICAL COMPOSITIONS AND ADMINISTRATION

Another embodiment provides pharmaceutical compositions or medicaments containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments. In one example, compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) is formulated in an acetate buffer, at pH 5. In another embodiment, the compounds of formula (I) are sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.

Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The “effective amount” of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to inhibit mutant RAS (e.g. KRAS G12C) interaction with RAF, blocking the oncogenic MAPK signaling. For example, such amount may be below the amount that is toxic to normal cells, or the mammal as a whole.

In one example, the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.1 to 1000 mg/kg, alternatively about 0.1 to 1000 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day. In another embodiment, oral unit dosage forms, such as tablets and capsules, preferably contain from about 1 to about 1000 mg of the compound of the invention.

The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.

A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).

An example of a suitable oral dosage form is a tablet containing about 1 to 1000 mg of the compound of the invention compounded with about 1 to 1000 mg anhydrous lactose, about 1 to 1000 mg sodium croscarmellose, about 1 to 1000 mg polyvinylpyrrolidone (PVP) K30, and about 1 to 1000 mg magnesium stearate. The powdered ingredients are first mixed together and then mixed with a solution of the PVP. The resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound, for example 5 to 400mg, of the invention in a suitable buffer solution, e.g. a phosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride, if desired. The solution may be filtered, e.g., using a 0.2 micron filter, to remove impurities and contaminants.

An embodiment, therefore, includes a pharmaceutical composition comprising a compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof. In a further embodiment includes a pharmaceutical composition comprising a compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier or excipient.

Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in the treatment of mutant KRAS-driven cancers. Another embodiment includes a pharmaceutical composition comprising a compound of Formula (I) for use in the treatment of mutant KRAS-driven cancers.

The following example A and B illustrate typical compositions of the present invention, but serve merely as representative thereof.

Example A

A compound of the present invention can be used in a manner known per se as the active ingredient for the production of tablets of the following composition:

Per tablet

Active ingredient 200 mg

Microcrystalline cellulose 155 mg

Corn starch 25 mg

Talc 25 mg

Hydroxypropylmethylcellulose 20 mg

425 mg

Example B

A compound of the present invention can be used in a manner known per se as the active ingredient for the production of capsules of the following composition:

Per capsule

Active ingredient 100.0 mg

Corn starch 20.0 mg

Lactose 95.0 mg

Talc 4.5 mg

Magnesium stearate 0.5 mg

220.0 mg

INDICATIONS AND METHODS OF TREATMENT The compounds of the invention induce a new binding pocket in KRAS by driving formation of a high affinity tri-complex between KRAS protein and the widely expressed cyclophilin A (CYPA), which inhibit KRAS interaction with downstream effectors, such as RAF and PI3K. Accordingly, the compounds of the invention are useful for inhibiting the propagating oncogenic MAPK and PI3K signaling, reducing cell proliferation, in particular cancer cells. Compounds of the invention are useful for termination of RAS signaling in cells that express RAS mutant, e.g. KRAS mutation driven pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma ovarian cancer, endometrial cancer, etc. Alternatively, compounds of the invention are useful for termination of RAS signaling in malignant solid tumor where the oncogenic role of KRAS mutation is reinforced by dysregulation or mutation of effector pathways as MAPK, PI3K-AKT-mT0R (Mammalian target of rapamycin) driven signaling, for targeted therapy in pancreatic adenocarcinoma, colorectal cancer, non- small cell lung cancer, etc.

Another embodiment includes a method of treating or preventing cancer in a mammal in need of such treatment, wherein the method comprises administering to said mammal a therapeutically effective amount of a compound of formula (I), a stereoisomer, tautomer, prodrug or pharmaceutically acceptable salt thereof.

SYNTHESIS

The compounds of the present invention can be prepared by any conventional means. Suitable processes for synthesizing these compounds as well as their starting materials are provided in the schemes below and in the examples. All substituents, in particular, R ¹ to R ⁷ , A ¹ and A ² are as defined above unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to a person of ordinary skill in organic chemistry.

General synthetic routes for preparing the compound of formula (I) are shown below.

Scheme 1

Compound of formula II was synthesized according to the procedure described in Intermediate A to L. Compound of formula (I) can be obtained by a coupling reaction between acid (III) and compound of formula (II) with coupling reagent(s), such as T3P, HATU, PyBOP and EDCI/HOBt, in the presence of a base, such as TEA, DIEPA and DMAP.

Scheme 2 wherein PG is a protecting group, such as Boc and Cbz. Alternatively compound of formula (V) can be obtained by a coupling reaction between acid (IV) and compound of formula (II) with coupling reagent(s), such as T3P, HATU, PyBOP or EDCI/HOBt, in the presence of a base, such TEA, DIEPA or DMAP. Deprotection of compound of formula (V) can afford compound of formula (VI) in the presence of an acid, such as TFA, or under hydrogenation condition with a catalyst, such as Pd/C and Pd(OH)2/C.

Compound of formula (VIII) can be obtained by a coupling reaction between acid (VII) and compound of formula (VI) with coupling reagent(s), such as T3P, HATU, PyBOP and EDCI/HOBt, in the presence of a base, such as TEA, DIEPA and DMAP.

Scheme 3 wherein PG is a protecting group, such as Boc and Cbz; T is dihaloC1-6alkyl; Q is unsubstituted or substituted piperidinylene or pyrrolidinylene.

Alternatively compound of formula (X) can be obtained by a coupling reaction using acid (IX), compound of formula (VI) and coupling reagent(s), such as T3P, HATU, PyBOP or EDCI/HOBt, in the presence of a base, such TEA, DIEPA or DMAP. Deprotection of compound of formula (X) can afford compound of formula (XI) in the presence of an acid, such as TFA, or under hydrogenation condition with a catalyst, such as Pd/C and Pd(OH)2/C. Compound of formula (XIII) can be obtained by a coupling reaction between acid (XII) and compound of formula (XI) with coupling reagent(s), such as T3P, HATU, PyBOP and EDCI/HOBt, in the presence of a base, such as TEA, DIEPA and DMAP.

Compounds of this invention can be obtained as mixtures of diastereomers or enantiomers, which can be separated by methods well known in the art, e.g. (chiral) HPLC or SFC. In another embodiment, compound of formula (I) can be obtained according to above scheme by using corresponding chiral starting materials.

This invention also relates to a process for the preparation of a compound of formula (I) comprising any of the following steps: a) coupling reaction between compound of formula (II), in the presence of a coupling reagent and a base to form the compound of formula (I); b) coupling reaction between compound of formula (VI), and acid (VII), (VII), in the presence of a coupling reagent and a base to form the compound of formula (VIII), c) coupling reaction between compound of formula (XI), and acid (XII), (XII), in the presence of a coupling reagent and a base to form the compound of formula (XIII), wherein in step a),b) and c) the coupling reagent can be, for example, T3P, HATU, PyBOP or EDCI/HOBt; the base can be, for example, TEA, DIEPA or DMAP; T is dihaloC1-6alkyl; Q is unsubstituted or substituted piperidinylene or pyrrolidinylene.

A compound of formula (I) or (la) when manufactured according to the above process is also an object of the invention.

EXAMPLES

The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.

ABBREVIATIONS

The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.

Abbreviations used herein are as follows:

ACN acetonitrile aq. Aqueous

Boc-A-Me-Val-OH A-(tert-Butoxycarbonyl)-A-methyl-L- valine

(BOC) ₂ O Di-terZ-butyldicarbonate

(7?)-binap (7?)-(+)-2,2'-Bis(diphenylphosphino)-l,r-binaphthyl CDC1 ₃ : deuterated chloroform

CD3OD: deuterated methanol

COMU ( 1 -Cyano -2-ethoxy-2-oxoethylidenaminooxy)dimethylamino- morpholino -carbenium hexafluoropho sphate

DIEPA: N, A-dicthylpropylaminc

DIBAL-H Diisobutylaluminium hydride

DMAP: 4-Dimethylaminopyridine

DMF: dimethyl formamide

DMSO: dimethyl sulfoxide

EDCI: N-Ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride

EtOAc or EA: ethyl acetate

FRET fluorescence resonance energy transfer

HATU: ( 1 - [B is(dimethy lamino ) methylene] - 1 H- 1 ,2 , 3 -triazolo [4,5- b]pyridinium 3-oxid hexafluoropho sphate) hr(s): hour(s)

HPLC: high performance liquid chromatography

HOBt: A-hydroxybenzotriazole

H-VAL-OTBU HC1 (S)-tert-Butyl 2-amino-3-methylbutanoate hydrochloride

[Ir(OMe)(COD)] ₂ ( 1 ,5-Cyclooctadiene)(methoxy)iridium(I) dimer

LDA Lithium diisopropylamide

MS: (ESI): mass spectroscopy (electron spray ionization) min(s) minute(s)

NMM N- Methylmorpholine

NMR: nuclear magnetic resonance

NMO 4-Methylmorpholine A-oxidc obsd. Observed

Pd(dppf)CI ₂ [1,1 '-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)

Pd(dtbpf)Cl ₂ [l,r-Bis(di-tert-butylphosphino)ferrocene]dichloropalladium( II) prep-HPLC preparative high performance liquid chromatography PyBOP: benzotriazol- 1-yloxytripyrrolidinophosphonium hexafluoropho sphate

RT or rt: room temperature sat. saturated SFC supercritical fluid chromatography

TEA: triethylamine

TFA: trifluoro acetic acid

THF: tetrahydro furan

TEA: triethylamine

T3P: propylphospho nic anhydride

GENERAL EXPERIMENTAL CONDITIONS

Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and the Quad 12/25 Cartridge module, ii) ISCO combi-flash chromatography instrument. Silica gel brand and pore size: i) KP-SIL 60 A, particle size: 40-60 pm; ii) CAS registry NO: Silica Gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Haiyang Chemical Co., Ltd, pore: 200-300 or 300-400.

Intermediates and final compounds were purified by preparative HPLC on reversed phase column using XBridge™ Prep-C18 (5 pm, OBDTM 30 x 100 mm) column, SunFire™ Prep-C18 (5 pm, OBD™ 30 x 100 mm) column, Phenomenex Synergi-C18 (10 pm, 25 x 150 mm) or Phenomenex Gemini-C18 (10 pm, 25 x 150 mm). Waters AutoP purification System (Sample Manager 2767, Pump 2525, Detector: Micromass ZQ and UV 2487, solvent system: acetonitrile and 0.1% ammonium hydroxide in water; acetonitrile and 0.1% FA in water or acetonitrile and 0.1% TFA in water). Or Gilson-281 purification System (Pump 322, Detector: UV 156, solvent system: acetonitrile and 0.05% ammonium hydroxide in water; acetonitrile and 0.225% FA in water; acetonitrile and 0.05% HC1 in water; acetonitrile and 0.075% TFA in water; or acetonitrile and water).

For SFC chiral separation, intermediates were separated by chiral column (Daicel chiralpak IC, 5 pm, 30 x 250 mm), AS (10 pm, 30 x 250 mm) or AD (10 pm, 30 x 250 mm) using Mettler Toledo Multigram III system SFC, Waters 80Q preparative SFC or Thar 80 preparative SFC, solvent system: CO ₂ and IPA (0.5% TEA in IPA) or CO ₂ and MeOH (0.1% NH3 H2O in MeOH), back pressure lOObar, detection UV @ 254 or 220 nm.

LC/MS spectra of compounds were obtained using a LC/MS (Waters™ Alliance 2795- Micromass ZQ, Shimadzu Alliance 2020-Micromass ZQ or Agilent Alliance 6110-Micromass ZQ), LC/MS conditions were as follows (running time 3 or 1.5 mins):

Acidic condition I: A: 0.1% TFA in H2O; B: 0.1% TFA in acetonitrile;

Acidic condition II: A: 0.0375% TFA in H2O; B: 0.01875% TFA in acetonitrile; Basic condition I: A: 0.1% NH3 H2O in H2O; B: acetonitrile;

Basic condition II: A: 0.025% NH3 H2O in H2O; B: acetonitrile;

Neutral condition: A: H2O; B: acetonitrile.

Mass spectra (MS): generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (MH) ⁺ .

NMR Spectra were obtained using Bruker Avance 400 MHz.

The microwave assisted reactions were carried out in a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were performed under an argon or nitrogen atmosphere. Reagents were used as received from commercial suppliers without further purification unless otherwise noted.

PREPARATIVE EXAMPLES

Preparation of Intermediate

Intermediate A l-[6-[(lS)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2-diox aborolan-2-yl)-3-pyridyl]-4- methyl-piperazine

The title intermediate A was prepared according to the following scheme:

Step 1: Preparation of 3-bromo-2-[(lS)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2 - dioxaborolan-2-yl)pyridine (compound A2)

To a solution of 3-bromo-2-[(15)-l-methoxyethyl]pyridine (compound Al, 2.0 g, 9.26 mmol) and bis(pinacolato)diboron (3.5 g, 13.9 mmol) in THF (30 mL) were added 4,4'-di-terZ- butyl-2,2'-bipyridin (372.7 mg, 1.39 mmol) and [Ir(OMe)(COD)]2 (306.3 mg, 0.460 mmol). The mixture was stirred at 75 °C for 16 hours under N2 protection. The mixture was filtrated and the filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (EA/PE: 0-20%) to afford 3-bromo-2-[(15)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2 - dioxaborolan-2-yl)pyridine (compound A2, 2.4 g) as yellow oil. ^J H NMR (400 MHz, CDC13) 6 ppm 8.91 (d, J = 1.4 Hz, 1 H), 8.21 (d, J = 1.4 Hz, 1 H), 4.95 (q, J = 6.5 Hz, 1 H), 3.30 (s, 3 H), 1.49 (d, J = 6.5 Hz, 3 H), 1.35 (s, 12 H).

Step 2: Preparation of 3-bromo-5-iodo-2-[(lS)-l-methoxyethyl]pyridine (compound A3) To a solution of 3-bromo-2-[(15)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2 - dioxaborolan-2-yl)pyridine (compound A2, 2.5 g, 7.3 mmol) in ACN (40 mL) was added N- iodo succinimide (4.1 g, 18.27 mmol). The mixture was stirred at 90 °C for 40 hrs under N2 protection. The reaction was quenched with saturated solution of Na2SO3 (40 mL) and the reaction mixture was extracted with EtOAc (30 mL, twice). The combined organic layer was washed with brine (50 mL), filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel chromatography (EA/PE: 0-20%) to afford 3-bromo-5-iodo-2-[(15)-l- methoxyethyl]pyridine (compound A3, 660 mg) as yellow oil. MS calc’d 342 (MH ⁺ ), measured 341.8 (MH ⁺ ).

Step 3: Preparation of benzyl 4-[5-bromo-6-[(lS)-l-methoxyethyl]-3- pyridyl]piperazine-l-carboxylate (Intermediate A5)

To a solution of 3-bromo-5-iodo-2-[(15)-l-methoxyethyl]pyridine (compound A3, 660 mg, 1.9 mmol) and 1-Cbz-piperazine (compound A4, 425.1 mg, 1.9 mmol) in toluene (10 mL) were added cesium carbonate (1.6 g, 4.83 mmol), (7?)-BINAP (60.1 mg, 0.1 mmol) and palladium (II) acetate (43.3 mg, 0.19 mmol). The mixture was stirred at 100 °C for 12 hours under N2 protection. The mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel chromatography (EA/PE: 0-50%) to afford benzyl 4-[5-bromo-6-[(lS)- l-methoxyethyl]-3-pyridyl]piperazine-l-carboxylate (Intermediate A5, 740 mg) as a yellow solid. MS calc’d 434.1 (MH ⁺ ), measured 434.1 (MH ⁺ ).

Step 4: Preparation of l-[6-[(lS)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-3-pyridyl]-4-methyl-piperazine (Intermediate A)

To a solution of benzyl 4-[5-bromo-6-[(15)-l-methoxyethyl]-3-pyridyl]piperazine-l- carboxylate (Intermediate A5, 740 mg, 1.7 mmol) and bis(pinacolato)diboron (519.2 mg, 2.04 mmol) in toluene (12 mL) were added KOAc (418.0 mg, 4.26 mmol) and Pd(dppf)Ch (124.7 mg, 0.170 mmol). The reaction mixture was stirred at 90 °C for 12 hrs under N2 protection. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by silica gel column to afford l-[6-[(15)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-3-pyridyl]-4-methyl-piperazine (Intermediate A, 470 mg) as a brown solid. MS calc’d 482.3 (MH ⁺ ), measured 482.2 (MH ⁺ ).

Intermediate B

Methyl (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetr amethyl-l,3,2- dioxaborolan-2-yl)-5-triisopropylsilyloxy-phenyl]propanoyl]h exahydropyridazine-3- carboxylate

The intermediate B was prepared according to the following scheme:

Step 1: Preparation of methyl (2S)-2-amino-3-(3-hydroxyphenyl)propanoate (compound B2)

To a solution of L-M-tyrosine (compound Bl, 5.0 g, 27.6 mmol) in methanol (80 mL) was added thionyl chloride (10 mL, 137.9 mmol). The mixture was stirred at 60 °C for 12 hrs. The reaction mixture was cooled to 20 °C and concentrated in vacuo to afford methyl (2S)-2-amino- 3-(3-hydroxyphenyl)propanoate (compound B2, 6.2 g) as a yellow solid. ^J H NMR (400 MHz, CD3OD) 8 ppm 7.18 (t, J = 8.0 Hz, 1H), 6.78 - 6.66 (m, 3H), 4.29 (t, J = 6.4 Hz, 1H), 3.82 (s, 3H), 3.23 - 3.05 (m, 2H).

Step 2: Preparation of methyl (2S)-2-(tert-butoxycarbonylamino)-3-(3- hydroxyphenyl)propanoate (compound B3)

To a solution of methyl (2S)-2-amino-3-(3-hydroxyphenyl)propanoate (compound B2, 32.0 g, 138.1 mmol) in THF (80 mL) and water (20 mL) was added sodium bicarbonate (40.6 g, 483.4 mmol) followed by di-Z-butyldicarbonate (33.1 g, 151.9 mmol) at 20 °C. The mixture was stirred at 20 °C for 12 hours. The mixture was diluted with water (100 mL) and acidified by 1 M aq. solution of HC1 until the pH=5. The mixture was extracted with ethyl acetate (100 mL, 3 times). The combined organic phase was washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum to afford methyl (25)-2- (tert-butoxycarbonylamino)-3-(3-hydroxyphenyl)propanoate (compound B3, 40 g) as colorless gum. MS: calc’d 318 (MNa ⁺ ), measured 318.3 (MNa ⁺ ).

Step 3: Preparation of methyl (2S)-2-(tert-butoxycarbonylamino)-3-(3- triisopropylsilyloxyphenyl)propanoate (compound B4)

To a solution of methyl (2S)-2-(tert-butoxycarbonylamino)-3-(3- hydroxyphenyl)propanoate (compound B3, 40.0 g, 135.4 mmol) and 1 H- imidazole (27.6 g, 406.3 mmol) in DMF (400 mL) was added triisopropylsilyl chloride (39.1 g, 203.1 mmol) dropwise at 0 °C. After being stirred for 12 hrs at 25 °C, the mixture was diluted with water (250 mL) at 0 °C and extracted with ethyl acetate (200 mL, 3 times). The combined organic phase was washed with brine (80 mL, four times), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum to give a residue. The residue was purified by silica gel chromatography (EA/PE: 0-20%) to afford methyl (2S)-2-(/c/7-butoxycarbonylamino)-3-(3- triisopropylsilyloxyphenyl)propanoate (compound B4, 60 g) as yellow oil. MS: calc’d 474 (MNa ⁺ ), measured 474.2 (MNa ⁺ ).

Step 4: Preparation of methyl (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-5-triisopropylsilyloxy- phenyl]propanoate (compound B5)

To a solution of methyl (2S)-2-(tert-butoxycarbonylamino)-3-(3- triisopropylsilyloxyphenyl)propanoate (compound B4, 15.0 g, 33.2 mmol), 4,4'-di-terZ-butyl- 2,2'-bipyridine (2.6 g, 9.9 mmol) and bis(pinacolato)diboron (12.6 g, 49.8 mmol) in hexane (200 mL) was added [Ir(OMe)(COD)]2 (2.2 g, 3.3 mmol). The mixture was degassed and purged with N2 for 3 times. The resulting mixture was stirred at 70 °C for 12 hrs. Then the reaction mixture was cooled to 20 °C, diluted with petroleum ether (100 mL) and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified by silica gel chromatography (EA/PE: 0-20%) to afford methyl (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-

1.3.2-dioxaborolan-2-yl)-5-triisopropylsilyloxy-phenyl]pr opanoate (compound B5, 21 g) as yellow oil. MS: calc’d 600 (MNa ⁺ ), measured 600.3 (MNa ⁺ ).

Step 5: Preparation of (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-

1.3.2-dioxaborolan-2-yl)-5-triisopropylsilyloxy-phenyl]pr opanoic add (compound B6)

To a solution of methyl (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl- l,3,2- dioxaborolan-2-yl)-5-triisopropylsilyloxy-phenyl]propanoate (compound B5, 40.0 g, 69.2 mmol) in methanol (300 mL) was added a solution of lithium hydroxide (3.2 mL, 346.2 mmol) in water (100 mL). After being stirred at 20 °C for 1 hour, the reaction mixture was diluted with water (200 mL) and MeOH was removed under vacuum. The resulting mixture was acidified by 1 M aq. solution of HC1 until the pH=5. The resulting mixture was extracted with EtOAc (250 mL, 3 times). The organic phase was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated in vacuo to afford (2S)-2-(/c/7-butoxycarbonylamino)-3- [3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-5-triisopro pylsilyloxy-phenyl]propanoic acid (compound B6, 33 g) as a white solid. MS: calc’d 586 (MNa ⁺ ), measured 586.3 (MNa ⁺ ).

Step 6: Preparation of methyl (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-5-triisopropylsilyloxy- phenyl]propanoyl]hexahydropyridazine-3-carboxylate (Intermediate B)

To a solution of (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl- l,3,2- dioxaborolan-2-yl)-5-triisopropylsilyloxy-phenyl]propanoic acid (compound B6, 8.0 g, 14.1 mmol) and O-(7-azabenzotriazol-l-yl)-A,A,A',A'-tetramethyluronium hexafluorophosphate (5.6 g, 14.9 mmol) in DMF (100 mL) was added A,A-diisopropylethylamine (6.4 g, 49.6 mmol). The mixture was stirred at 0 °C for 10 min. Then methyl (3S)-hexahydropyridazine-3-carboxylate hydrochloride salt (compound B7, 2.6 g, 14.9 mmol) was added. The resulting mixture was stirred at 20 °C for 1.5 hrs. The mixture was diluted with water (200 mL) and extracted with EtOAc (100 mL, twice). The combined organic phase was washed with brine, dried over anhydrous sodium sulfate, filtrated and the filtrate was concentrated under vacuum. The residue was purified by silica gel chromatography to afford methyl (3S)-l-[(2S)-2-(tert- butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-l,3,2-dioxabo rolan-2-yl)-5- triisopropylsilyloxy-phenyl]propanoyl]hexahydropyridazine-3- carboxylate (intermediate B, 7.8 g) as yellow oil. MS: calc’d 690 (MH ⁺ ), measured 690.4 (MH ⁺ ).

Intermediate C

Methyl (3S)-l-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonyla mino)- propanoyl]hexahydropyridazine-3-carboxylate

The intermediate C was prepared according to the following scheme:

Step 1: Preparation of (4-bromothiazol-2-yl)methanol (compound C2)

To a solution of 4-bromothiazole-2-carboxaldehyde (6.0 g, 31.25 mmol) in methanol (70 mL) was added sodium borohydride (1.7 g, 46.87 mmol) at 0 °C. The mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (300 mL) at 0 °C and the reaction mixture was extracted by ethyl acetate (200 mL, three times). The combined organic phase was washed with brine (150 mL, twice), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum to afford (4-bromothiazol-2-yl)methanol (compound C2, 6g) as colorless oil. Step 2: Preparation of 4-bromo-2-(bromomethyl)thiazole (compound C3)

To a solution of (4-bromothiazol-2-yl)methanol (compound C2, 6.0 g, 30.92 mmol) in DCM (80 mL) was added CBr4 (15.4 g, 46.38 mmol) and triphenylphosphine (12.1 g, 46.38 mmol) at 0 °C. After being stirred at 25 °C for 1 hour, the mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column, eluted with ethyl acetate in petroleum ether = 0-10% to afford (4-bromothiazol-2-yl)methanol (compound C3, 6.0 g) as yellow oil. MS calc’d 255.9 (MH ⁺ ), measured 255.9 (MH ⁺ ).

Step 3: Preparation of 4-bromo-2-[[(2S,5/f)-5-isopropyl-3,6-dimethoxy-2,5- dihydropyrazin-2-yl]methyl]thiazole (compound C5)

To a mixture of (7?)-2,5-dihydro-3,6-dimethoxy-2-isopropylpyrazine (compound C4, 4.3 g, 23.45 mmol) in THF (60 mL) was added n-butyllithium (10 mL, 25.22 mmol, 2.5 M) at -78 °C slowly. After addition, the mixture was stirred for 0.5 hour at -78 °C. 4-bromo-2- (bromomethyl)thiazole (compound C3, 5.4 g, 21.02 mmol) was added into above mixture at - 78 °C which was stirred for another 1 hour. The reaction was quenched with saturated solution of NH4CI (100 mL) and the reaction mixture was extracted with EtOAc (100 mL, twice). The combined organic layer was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum. The residue was purified by reversed- phase chromatography to afford 4-bromo-2-[[(2S,57?)-5-isopropyl-3,6-dimethoxy-2,5- dihydropyrazin-2-yl]methyl]thiazole (compound C5, 3.6 g) as yellow oil. MS calc’d 360 (MH ⁺ ), measured 359.9 (MH ⁺ ).

Step 4: Preparation of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (compound C6)

To a solution of 4-bromo-2-[[(2S,57?)-5-isopropyl-3,6-dimethoxy-2,5-dihydropy razin-2- yl] methyl] thiazole (compound C5, 3.6 g, 10 mmol) in ACN (20 mL) was added hydrochloric acid (66.6 mL, 0.3 M). The mixture was stirred at 25 °C for 2 hours. The mixture was basified by saturated solution of NaHCCL until pH=8. The mixture was extracted with EtOAc (80 mL, six times). The combined organic layer was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum to afford methyl (2S)-2-amino-3-(4-bromothiazol-2- yl)propanoate (compound C6, 3.1 g) as yellow oil. MS calc’d 264.9 (MH ⁺ ), measured 264.9 (MH ⁺ ).

Step 5: Preparation of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert- butoxycarbonylamino)propanoate (compound C7)

To a solution of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (compound C6, 3.1 g, 11.69 mmol) in DCM (40 mL) were added triethylamine (2.9 g, 29.23 mmol) and (BochO (3.8 g, 17.54 mmol). After being stirred at 30 °C for 12 hours, the mixture was concentrated under vacuum. The residue was purified by silica gel column, eluted with ethyl acetate in petroleum ether = 0-30% to afford methyl (2S)-3-(4-bromothiazol-2-yl)-2-(/c/7- butoxycarbonylamino)propanoate (compound C7, 3.2 g) as yellow oil. MS calc’d 387(MNa ⁺ ), measured 386.9 (MNa ⁺ ).

Step 6: Preparation of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)- propanoic add (compound C8)

To a solution of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(terZ- butoxycarbonylamino)propanoate (compound C7, 3.2 g, 8.76 mmol) in THF (30 mL), methanol (2 mL) and water (10 mL) was added lithium hydroxide (0.4 mL, 43.81 mmol). After being stirred at 25 °C for 1 hour, the reaction mixture was acidified by 1 M solution of HC1 until pH=5. The mixture was extracted with EtOAc (40 mL, twice). The combined organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum to afford (2S)-3-(4-bromothiazol-2-yl)-2-(/c/7- butoxycarbonylamino)propanoic acid (compound C8, 3.1 g) as yellow oil. MS calc’d 373(MNa ⁺ ), measured 372.9 (MNa ⁺ ).

Step 7: Preparation of methyl (3S)-l-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ate (Intermediate C)

To a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(terZ-butoxycarbonylamino)pro panoic acid (compound C8, 3.1 g, 8.83 mmol) in DCM (50 mL) was added methyl (3S)- hexahydropyridazine-3-carboxylate;hydrochloride (compound C9, 2.4 g, 13.24 mmol), EDCI (3.4 g, 17.65 mmol), 1 -Hydroxybenzotriazole (238.5 mg, 1.77 mmol) and NMM (9.92 mL, 88.26 mmol) at 0 °C. After being stirred at 25 °C for 1 hour, the reaction mixture was diluted with water (60 mL) and extracted with EtOAc (60 mL, three times). The combined organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column and eluted with ethyl acetate in petroleum ether = 10-30% to afford methyl (3S)- l -[(2S)-3-(4-bromothiazol-2-yl)-2- (tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-ca rboxylate (intermediate C, 2.4 g). MS calc’d 477(MH ⁺ ), measured 476.9 (MH ⁺ ).

Intermediate D (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl ]-5-(4-methylpiperazin-l-yl)- 3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14- dione

The title intermediate D was prepared according to the following scheme:

D13 intermediate D

Step 1: Preparation of l-(5-bromo-6-fluoro-lH-indol-3-yl)-3-((tert-butyldiphenylsil yl) oxy)-2,2-dimethylpropan-l-one (compound D3) To a mixture of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (compound DI, 35.0 g, 116.8 mmol) in DCM (400 mL) at 0 °C was added a solution of SnCU (97.2 mL, 121.5 mmol) slowly. After being stirred at - 40 °C for 0.5 hour, 5-bromo-6-fluoro-l/Z- indole (compound D2, 25.0 g, 116.8 mmol) in DCM (200 mL) was added dropwise and the mixture was stirred at - 40 °C for 15 min. After the reaction was completed, it was quenched with sat.NaHCCh aq. (800 mL), and the reaction mixture was extracted with EtOAc (900 mL, twice). The combined organic layer was washed with brine (700 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was triturated with the solution (100 mL, Petroleum ether: Ethyl acetate = 8:1) and filtered. The filter cake was dried in vacuo to afford 1- (5-bromo-6-fluoro-l/Z-indol-3-yl)-3-((tertbutyldiphenylsilyl )oxy)-2,2-dimethylpropan-l-one (compound D3, 50.0 g) as a yellow solid. MS calc’d 552.1 (MH ⁺ ), measured 552.1 (MH ⁺ ).

Step 2: Preparation of [3-(5-bromo-6-fluoro- lH-indol-3-yl)-2,2-dimethyl-propoxy ]- /cr/-butyl-diphenyl-silane (compound D4)

To a mixture of l-(5-bromo-6-fluoro-l/Z-indol-3-yl)-3-((tertbutyldiphenylsil yl)oxy)-2,2- dimethylpropan-l-one (compound D3, 50.0 g, 90.49 mmol) in THF (600 mL) was added LiBPL (48.4 mL, 193.49 mmol, 4 M in THF) dropwise at 0 °C. The mixture was stirred at 70 °C for 24 hrs under nitrogen atmosphere. After the reaction was completed, it was quenched by addition of water (600 mL) at 0 °C slowly and the reaction mixture was extracted with EtOAc (600 mL, twice). The combined organic layer was washed with brine (600 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 20% ~ 33%) to afford [3-(5-bromo-6-fluoro-l/Z-indol-3-yl)-2,2-dimethyl- propoxy]-terZ-butyl-diphenyl-silane (compound D4, 46.0 g) as a white solid. MS calc’d 538.1 (MH ⁺ ), measured 538.2 (MH ⁺ ).

Step 3: Preparation of [3-(5-bromo-6-fluoro-2-iodo-lH-indol-3-yl)-2,2-dimethyl- propoxy ]-/cr/-butyl-diphenyl-silane (compound D5)

To a mixture of [3-(5-bromo-6-fluoro-l/Z-indol-3-yl)-2,2-dimethyl-propoxy]-t ert-butyl- diphenyl- silane (compound D4, 35.4 g, 65.73 mmol) and iodine (18.4 g, 72.3 mmol) in THF (400 mL) was added silver trifluoromethanesulfonate (20.3 g, 78.88 mmol) at 0 °C. The mixture was stirred at 0 °C for 10 min. After the reaction was completed, it was quenched by sat. Na2SO3 aq. (400 mL) and EtOAc (400 mL) and the reaction mixture was filtered. The organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 0% ~ 2.5%) to afford [3-(5- bromo-6-fluoro-2-iodo- 1/7- indo l-3-yl)-2,2-dimethyl-propoxy]-rert-butyl-diphenyl-silane (compound D5, 43.0 g) as a yellow solid. MS calc’d 664.0 (MH ⁺ ), measured 664.1 (MH ⁺ ).

Step 4: Preparation of benzyl 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2- dimethyl-propyl]-6-fluoro-lH-indol-2-yl]-6-[(lS)-l-methoxyet hyl]-3-pyridyl]piperazine-l- carboxylate (compound D6)

To a mixture of [3-(5-bromo-6-fluoro-2-iodo-l//-indol-3-yl)-2,2-dimethyl-pro poxy]-rert- butyl-diphenyl- silane (compound D5, 16.7 g, 25.13 mmol) and benzyl 4-[6-[(lS)-l- methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl )-3-pyridyl]piperazine-l- carboxylate (Intermediate A, 16.7 g, 34.69 mmol) in a mixed solution of 1,4-dioxane (270 mL)/Toluene (90 mL) /water (90 mL) were added potassium phosphate (15.7 g, 73.92 mmol) and Pd(dppf)Ch (920 mg, 1.26 mmol). The mixture was stirred at 70 °C for 12 hrs under nitrogen atmosphere. After the reaction was completed, the mixture was filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 20% ~ 50%) to afford 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimeth yl-propyl]- 6-fluoro-l//-indol-2-yl]-6-[(lS)-l-methoxyethyl]-3-pyridyl]p iperazine-l-carboxylate (compound D6, 19.5 g) as a white solid. MS calc’d 891.3 (MH ⁺ ), measured 891.3 (MH ⁺ ).

Step 5: Preparation of benzyl 4-|(5 V7)-5-|5-bronio-3-|3-|/cr/-butyl(diphenyl)silyl|oxy- 2,2-dimethyl-propyl]-6-fluoro-l-(2,2,2-trifluoroethyl)indol- 2-yl]-6-[(lS)-l-methoxyethyl]-3- pyridyl]piperazine-l-carboxylate(compound D7)

To a solution of 4-[5-[5-bromo-3-[3-[terZ-butyl(diphenyl)silyl]oxy-2,2-dimeth yl-propyl]-6- fluoro- 1H- indo l-2-yl]-6-[( IS)- l-methoxyethyl]-3-pyridyl]piperazine-l-carboxylate (compound D6, 14.5 g, 16.26 mmol) and CS2CO3 (15. 9 g, 48.77 mmol) in DMF (200 mL) was added 2,2,2- trifluoroethyl trifluoromethanesulfonate (37.7 g, 162.56 mmol) dropwise at 0 °C, and the mixture was stirred at 20 °C for 12 hrs. After the reaction was completed, EtOAc (70 mL) and water (100 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (70 mL, twice). Combined organic layer was washed with brine (100 mL, four times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica column chromatography to afford benzyl 4-[(5A7)-5-[5-bromo-3-[3-[tert- butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-l-(2, 2,2-trifluoroethyl)indol-2-yl]-6- [(lS)-l-methoxyethyl]-3-pyridyl]piperazine-l-carboxylate (compound D7, 8.0 g, PEAK 1, faster eluted) as yellow oil. MS calc’d 973.3 (MH ⁺ ), measured 973.2 (MH ⁺ ). Step 6: Preparation of benzyl 4-[(5Af)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2- dimethyl-propyl)-l-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(lS) -l-methoxyethyl]-3- pyridyl]piperazine-l-carboxylate (compound D8)

To a solution of benzyl 4-[(5M)-5-[5-bromo-3-[3-[terZ-butyl(diphenyl)silyl]oxy-2,2- dimethyl-propyl]-6-fluoro-l-(2,2,2-trifluoroethyl)indol-2-yl ]-6-[(lS)-l-methoxyethyl]-3- pyridyl]piperazine-l -carboxylate (compound D7, 10.5 g, 10.78 mmol) in DMF (130 mL) was added cesium fluoride (8.2 g, 53.9 mmol) and the mixture was stirred at 60 °C for 24 hrs. After the reaction was completed, EtOAc (100 mL) and water (100 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (100 mL, twice). The combined organic layer was washed with brine (80 mL, three times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica column chromatography (EtOAc in PE = 25% ~ 66%) to afford benzyl 4-[(5A7)-5-[5-bromo-6-fluoro-3- (3-hydroxy-2,2-dimethyl-propyl)-l-(2,2,2-trifluoroethyl)indo l-2-yl]-6-[(15)-l-methoxyethyl]-3- pyridyl]piperazine-l -carboxylate (compound D8, 6.5 g) as a yellow solid. MS calc’d 735.2 (MH ⁺ ), measured 735.1 (MH ⁺ ).

Step 7: Preparation of benzyl 4-[(5Af)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)- 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l-(2,2,2-tri fluoroethyl)indol-2-yl]-6-[(lS)-l- methoxyethyl]-3-pyridyl]piperazine-l-carboxylate (compound D9)

To a solution of benzyl 4-[(5A7)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propy l)- l-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(lS)-l-methoxyethyl]- 3-pyridyl]piperazine-l-carboxylate (compound D8, 5.4 g) , bis(pinacolato)diboron (2.8 g, 11.01 mmol) and potassium acetate (1.2 mL, 18.35 mmol) in Toluene (70 mL) was added Pd(dppf)Ch (537.1 mg, 0.73 mmol). The mixture was degassed and purged with nitrogen atmosphere for three times and the mixture was stirred at 90 °C for 12 hrs. After the reaction was completed, the mixture was cooled to room temperature. The reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by silica column chromatography (EtOAc in PE = 25% ~ 66%) to afford benzyl 4-[(5A7)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-l-(2,2,2-trifluoroethyl )indol-2-yl]-6-[(15)-l- methoxyethyl]-3-pyridyl]piperazine-l-carboxylate (compound D9, 5.2 g) as yellow oil. MS calc’d 783.3 (MH ⁺ ), measured 783.3 (MH ⁺ ).

Step 8: Preparation of methyl (3S)-l-[(2S)-3-[4-[(2M)-2-[5-(4- benzyloxycarbonylpiperazin-l-yl)-2-[(lS)-l-methoxyethyl]-3-p yridyl]-6-fluoro-3-(3- hydroxy-2,2-dimethyl-propyl)-l-(2,2,2-trifluoroethyl)indol-5 -yl]thiazol-2-yl]-2-(tert- butoxycarbonylamino)-propanoyl]hexahydropyridazine-3-carboxy late (compound DIO)

To a mixture of methyl (3S)-l-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ate (intermediate C, 2.7 g, 5.69 mmol), benzyl 4-[(5A7)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-l-(2,2,2-trifluoroethyl )indol-2-yl]-6-[(15)-l- methoxyethyl]-3-pyridyl]piperazine-l-carboxylate (compound D9, 4.9 g, 6.32 mmol) in toluene (60 mL)/l,4-dioxane (20 mL) / water (20 mL) were added K3PO4 (3.4 g, 15.81 mmol) and Pd(dtbpf)Ch (412.2 mg, 0.63 mmol) under nitrogen atmosphere. The mixture was stirred at 70 °C for 12 hrs. After the reaction was completed, the mixture was concentrated in vacuo to give a residue. The residue was purified by silica column (EtOAc in PE = 10% ~ 75%) to afford methyl (3S)-l-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-l -yl)-2-[(15)-l-methoxyethyl]-3- pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-l-(2,2,2 -trifluoroethyl)indol-5-yl]thiazol-2- yl]-2-(tert-butoxycarbonylamino)-propanoyl]hexahydropyridazi ne-3-carboxylate (compound D10, 3.6 g) as a brown solid. MS calc’d 1053.4 (MEE), measured 1053.3 (MH ⁺ ).

Step 9: Preparation of (3S)-l-[(2S)-3-[4-[(2Af)-2-[5-(4-benzyloxycarbonylpiperazin- l- yl)-2-[(lS)-l-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy -2,2-dimethyl-propyl)-l-(2,2,2- trifhioroethyl)indol-5-yl]thiazol-2-yl]-2-(/c/7-butoxycarbon ylamino)propanoyl]hexahy- dropyridazine-3-carboxylic add (compound Dll)

To a solution methyl (3S)-l-[(2S)-3-[4-[(2Af)-2-[5-(4-benzyloxycarbonylpiperazin- l-yl)-2- [(15)-l-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-d imethyl-propyl)-l-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbon ylamino)-propanoyl]- hexahydropyridazine-3-carboxylate (compound D10, 3.6 g, 3.42 mmol) in DCE (50 mL) was added trimethylstannanol (2.4 g, 13.67 mmol) and the mixture was stirred at 60 °C for 12 hrs. After the reaction was completed, EtOAc (80 mL) and water (60 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (80 mL, twice). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give (3S)-l-[(2S)-3-[4-[(2A7)-2-[5-(4-benzyloxycarbonylpiperazin- l-yl)-2- [(15)-l-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-d imethyl-propyl)-l-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbon ylamino)propanoyl]hexahy- dropyridazine-3-carboxylic acid (compound Dll, 4.3 g) as a brown solid. MS calc’d 1039.4 (MH ⁺ ), measured 1039.2 (MH ⁺ ). Step 10: Preparation of benzyl 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24- fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-1 5-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-(20Af)- 20-yl]-6-[(lS)-l-methoxyethyl]-3-pyridyl]piperazine-l-carbox ylate (compound D12)

To a mixture of (3S)- l -[(2S)-3-[4-[(2A7)-2-[5-(4-bcnzyloxycarbonylpipcrazin- l -yl)-2- [(15)-l-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-d imethyl-propyl)-l-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbon ylamino)propanoyl]hexahy- dropyridazine-3-carboxylic acid (compound Dll, 4.3 g, 4.14 mmol) in DCM (430 mL) was added DIEA (14.4 mL, 82.76 mmol), EDCI (11.9 g, 62.07 mmol) and 1 -hydroxybenzotriazole (1.4 g, 10.35 mmol) at 0 °C. The mixture was stirred at 15 °C for 12 hrs. After the reaction was completed, the mixture was concentrated in vacuo, then diluted with water (80 mL), extracted with EtOAc (80 mL, twice). The combined organic layer was washed with brine (80 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 25% ~ 66%) to afford benzyl 4-[5-[(7S,13S)-7-(tert- butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21- (2,2,2-trifluoroethyl)-15-oxa-4- thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-(20A7)-20-yl]-6-[(lS)-l-methoxyethyl]-3-pyridyl]piper azine-l-carboxylate (compound D12, 3.1 g) as yellow gum. MS calc’d 1021.4 (MH ⁺ ), measured 1021.2 (MH ⁺ ).

Step 11: Preparation of tett-butyl A^-[(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamate (compound D13)

To a mixture of benzyl 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-(20A7)-20- yl]-6-[(15)-l-methoxyethyl]-3-pyridyl]piperazine-l-carboxyla te (compound D12, 3.1 g, 3.04 mmol) and formaldehyde aqueous (775.0 mg, 9.55 mmol) in methanol (150 mL) was added Pd(OH)2 on activated carbon (2.79 g, 3.97 mmol). The mixture was degassed and purged with H2 three times. The mixture was hydrogenated at 30 °C for 18 hrs. After the reaction was completed, the mixture was filtered and the filtrate was concentrated in vacuo to afford tert-butyl N- [(7S, L3S)-24-fluoro-(20A7)-20-[2-[( lS)- l-mcthoxycthyl]-5-(4-mcthylpipcrazin- l -yl)-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamate (compound D13, 2.6 g) as a brown solid. MS calc’d 901.3 (MH ⁺ ), measured 901.3 (MH ⁺ ).

Step 12: Preparation of (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D)

To a mixture of /c/7-butyl A ^/ -[(7SJ 3S)-24-fluoro-(20A7)-20-[2-[( lS)- l -mcthoxycthyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-2 1-(2,2,2-trifluoroethyl)-15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl] carbamate (compound D13, 2.6 g, 2.89 mmol) in DCM (18 mL) was added TFA (14.0 mL, 181.72 mmol). The mixture was stirred at 15 °C for 0.5 h. After the reaction was completed, the mixture was concentrated in vacuo and diluted with sat. NaHCO? (30 mL), extracted with EtOAc (30 mL, three times). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford (7S,13S)-7-amino-24- fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D, 2.0 g) as a yellow solid, which was used directly in the next step. MS calc’d 801.3 (MH ⁺ ), measured 801.2 (MH ⁺ )

Intermediate E

(7S,13S)-7-amino-25-fluoro-(20Af)-20-[2-[(lS)-l-methoxyet hyl]-5-(4-methylpiperazin-l-yl)- 3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14- dione

The title compound was prepared in analogy to the preparation of Intermediate D by using

5-bromo-4-fluoro- 177- indole instead of 5-bromo-6-fluoro-177-indole (compound D2).

Intermediate F

(7S,13S)-7-amino-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]- 17,17,24-trimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9 ,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14- dione

The title compound was prepared in analogy to the preparation of Intermediate D by using 5-bromo-6-methyl- 177- indole instead of 5-bromo-6-fluoro-177-indole (compound D2).

Intermediate G (8S,14S)-8-amino-25-fluoro-4-hydroxy-(21Af)-21-[2-[(lS)-l-me thoxyethyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl|- 18, 18-dimethyl-22-(2,2, 2-trifluoroethyl)- 16-oxa- 10.22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ¹⁰ ’ ¹⁴ .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaene-9,15- dione

The compound Intermediate G was prepared according to the following scheme:

Step 1: Preparation of methyl (3S)-l-[(2S)-3-[3-[(2M)-2-[5-(4- benzyloxy carbonylpiperazin-l-yl)-2-[(lS)-l-methoxyethyl]-3-pyridyl]-6 -fluoro-3-(3- hydroxy-2,2-dimethyl-propyl)-l-(2,2,2-trifluoroethyl)indol-5 -yl]-5-triisopropylsilyloxy- phenyl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyrid azine-3-carboxylate (compound Gl)

To a mixture of benzyl 4-[(5A7)-5-[5-bromo-l-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimeth yl- propyl)indol-2-yl]-6-[(15)-l-methoxyethyl]-3-pyridyl]piperaz ine-l-carboxylate (compound D8, 950.0 mg, 1.29 mmol) and methyl (35)-l-[(25)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5- tetramethyl- 1,3, 2-dioxaboro lan-2-yl)-5-triisopropylsilylo xy- phenyl]propanoyl]hexahydropyridazine-3-carboxylate (intermediate B, 1.07 g, 1.55 mmol) in 1,4-dioxane (25 mL) and water (5 mL) were added sodium carbonate (342.2 mg, 3.23 mmol) and Pd(dtbpf)Ch (84.2 mg, 0.13 mmol). The mixture was stirred at 85 °C for 12 hrs under nitrogen atmosphere. After the reaction was completed, the mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by reversed phase flash column and concentrated in vacuo to afford methyl (3S)-l-[(2S)-3-[3-[(2Af)-2-[5-(4- benzyloxycarbonylpiperazin-l-yl)-2-[(lS)-l-methoxyethyl]-3-p yridyl]-6-fluoro-3-(3-hydroxy- 2,2-dimethyl-propyl)-l-(2,2,2-trifluoroethyl)indol-5-yl]-5-t riisopropylsilyloxy-phenyl]-2-(tert- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ate (compound Gl, 1.3 g) as a brown solid. MS calc’d 1218.6 (MH ⁺ ), measured 1218.5 (MH ⁺ ).

Step 2: Preparation of (3S)-l-[(2S)-3-[3-[(2Af)-2-[5-(4-benzyloxycarbonylpiperazin- l- yl)-2-[(lS)-l-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy -2,2-dimethyl-propyl)-l-(2,2,2- trifhioroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2- (/('/7- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ic add (compound G2)

To a mixture of methyl (3S)-l-[(2S)-3-[3-[(2Af)-2-[5-(4-benzyloxycarbonylpiperazin- l-yl)- 2-[(15)-l-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2 -dimethyl-propyl)-l-(2,2,2- trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2- (tert-butoxycarbonylamino)- propanoyl]hexahydropyridazine-3-carboxylate (compound Gl, 1.1 g, 0.9 mmol) in DCE (20 mL) was added MesSnOH (652.9 mg, 3.61 mmol). The mixture was stirred at 60 °C for 12 hrs. After the reaction was completed, the mixture was diluted with water (100 mL), extracted with EtOAc (100 mL, twice). The combined organic layer was washed with brine (150 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford (3S)-l-[(2S)-3-[3-[(2Af)-2-[5-(4- benzyloxycarbonylpiperazin-l-yl)-2-[(lS)-l-methoxyethyl]-3-p yridyl]-6-fluoro-3-(3-hydroxy- 2,2-dimethyl-propyl)-l-(2,2,2-trifluoroethyl)indol-5-yl]-5-t riisopropylsilyloxy-phenyl]-2-(tert- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ic acid (compound G2, 1.5 g) as a yellow solid, which was used directly in the next step. MS calc’d 1204.6 (MH ⁺ ), measured 1204.5 (MH ⁺ ).

Step 3: Preparation of benzyl 4-[5-[(8S,14S)-8-(tert-butoxycarbonylamino)-25-fluoro- 18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triiso propylsilyloxy-16-oxa-10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ¹⁰ ’ ¹⁴ .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaen- (21 17)-21 -yl|-6-| ( LS)- l-methoxyethyl|-3-pyridyl|piperazine- 1 -carboxylate (compound G3)

To a mixture of (3S)-l-[(2S)-3-[3-[(2A7)-2-[5-(4-benzyloxycarbonylpiperazin- l-yl)-2- [(15)-l-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-d imethyl-propyl)-l-(2,2,2- trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2- (tert-butoxycarbonylamino)- propanoyl]hexahydropyridazine-3-carboxylic acid (compound G2, 1.5 g, 1.25 mmol) in DCM (150 mL) were added DIEA (4.4 mL, 24.95 mmol), EDCI (3.6 g, 18.71 mmol) and HOBt (421.4 mg, 3.12 mmol) at 0 °C. After being stirred at 15 °C for 12 hrs, the reaction mixture was concentrated in vacuo and diluted with water (50 mL), extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (EtOAc in PE = 25% ~ 50%) to afford benzyl 4-[5-[(8S,14S)-8-(tert-butoxycarbonylamino)-25-fluoro- 18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triiso propylsilyloxy-16-oxa-10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaen-(21A7)- 21-yl]-6-[(15)-l-methoxyethyl]-3-pyridyl]piperazine-l-carbox ylate (compound G3, 1.1 g) as a yellow solid. MS calc’d 1186.4 (MH ⁺ ), measured 1186.5 (MH ⁺ ).

Step 4: Preparation oftert-butyl A^-[(8S,14S)-25-fluoro-(21Af)-21-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-18,18-di methyl-9,15-dioxo-22-(2,2,2- trifluoroethyl)-4-triisopropylsilyloxy-16-oxa-10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ¹⁰ ’ ¹⁴ .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaen-8- yl] carbamate (compound G4)

To a mixture of benzyl 4-[5-[(8S,14S)-8-(tert-butoxycarbonylamino)-25-fluoro-18,18- dimethyl-9, 15-dioxo-22-(2, 2, 2-trifluoroethyl)-4-triisopropylsilylo xy- 16-oxa- 10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaen-(21A7)- 21-yl]-6-[(15)-l-methoxyethyl]-3-pyridyl]piperazine-l-carbox ylate (compound G3, 890.0 mg, 0.75 mmol) and formaldehyde (182.6 mg, 2.25 mmol) in methanol (15 mL) was added Pd(OH)2 on activated carbon (700.0 mg, 1.0 mmol). The mixture was degassed and purged with H2 three times. The mixture was hydrogenated at 30 °C for 18 hrs. The mixture was filtered, the filtrate was concentrated in vacuo to afford tert-butyl A-[(8S,14S)-25-fluoro-(21Af)-21-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-18,18-di methyl-9,15-dioxo-22-(2,2,2- trifluoroethyl)-4-triisopropylsilyloxy- 16-oxa- 10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaen-8- yl]carbamate (compound G4, 780.0 mg) as colorless gum, which was used directly in the next step. MS calc’d 1066.5 (MH ⁺ ), measured 1066.7 (MH ⁺ ).

Step 5: Preparation oftert-butyl A^-[(8S,14S)-25-fluoro-4-hydroxy-(21Af)-21-[2-[(lS)- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-18,18- dimethyl-9,15-dioxo-22-(2,2,2-

Z 6 10 14 2327 trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ’ .1 ’ .0 ’ ]nonacosa- l(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (compound G5) To a mixture of afford tert-butyl A-[(8S,14S)-25-fluoro-(21Af)-21-[2-[(lS')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-18,18-di methyl-9,15-dioxo-22-(2,2,2- trifluoroethyl)-4-triisopropylsilyloxy- 16-oxa- 10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaen-8- yl]carbamate (compound G4, 780.0 mg, 0.73 mmol) in THF (10 mL) was added tetrabutylammonium fluoride (0.8 mL, 0.8 mmol, 1 M in THF) at 0 °C. After being stirred at 0 °C for 0.5 h, the reaction mixture was diluted with water (20 mL), extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (30 mL, twice), dried over Na2SO4, filtered and concentrated in vacuo to afford /c/7-butyl A-[(8S, 14S)-25-fluoro-4-hydroxy- (21A7)-21-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl )-3-pyridyl]-18,18-dimethyl-9,15- dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa-l(26),2,4,6(29),20,23(27),24-heptaen-8- yl]carbamate (compound G5, 700.0 mg, 0.77 mmol) as a yellow solid, which was directly used in the next step. MS calc’d 910.4 (MH ⁺ ), measured 910.7 (MH ⁺ ).

Step 6: Preparation of (8S,14S)-8-amino-25-fluoro-4-hydroxy-(21Af)-21-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-18,18-di methyl-22-(2,2,2-

Z 6 10 14 2327 trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ’ .1 ’ .0 ’ ]nonacosa- 1(26), 2, 4, 6(29), 20, 23(27), 24-heptaene-9, 15-dione (intermediate G)

To a mixture of tert-butyl A-[(8S,14S)-25-fluoro-4-hydroxy-(21A7)-21-[2-[(lS')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-18,18-di methyl-9,15-dioxo-22-(2,2,2- 2 10 14 23 27 trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 .1 ’ .0 ]nonacosa- l(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (compound G5, 700.0 mg, 0.77 mmol) in DCM (10 mL) was added TFA (6.0 mL, 77.88 mmol). After being stirred at 15 °C for 0.5 h, the reaction mixture was concentrated in vacuo and diluted with sat. NaHCCh (15 mL), extracted with EtOAc (50 mL, twice). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford (8S, 14S)-8-amino-25-fluoro-4- hydroxy-(21A7)-21-[2-[(lS)-l-methoxyethyl]-5-(4-methylpipera zin-l-yl)-3-pyridyl]-18,18- dimethyl-22-(2,2,2-trifluoroethyl)-16-oxa- 10,22,28- triazapentacyc lo[ 18.5.2.1 ² ’ ⁶ . l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa- 1(26), 2, 4, 6(29), 20, 23(27), 24-heptaene-9, 15- dione (intermediate G, 600.0 mg) as a yellow solid, which was used directly in the next step. MS calc’d 810.3 (MH ⁺ ), measured 810.4 (MH ⁺ ).

Intermediate H (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-met hoxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thi a-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaene-8,14- dione

The title compound was prepared in analogy to the preparation of Intermediate D by using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate.

Intermediate I (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl ]-5-morpholino-3-pyridyl]- 17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21, 27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14- dione

The compound was prepared according to the following scheme:

Intermediate I no Step 1: Preparation of 4-[5-bromo-6-[(lS)-l-methoxyethyl]-3-pyridyl]morpholine (compound 12)

To a mixture of 3-bromo-5-iodo-2-[(15)-l-methoxyethyl]pyridine (compound II, 3.4 g, 9.94 mmol) and morpholine (0.9 g, 9.94 mmol) in Toluene (50 mL) were added cesium carbonate (8.1 g, 24.87 mmol), (7?)-binap (0.3 g, 0.5 mmol) and palladium (II) acetate (0.1 g, 0.5 mmol). The mixture was stirred at 100 °C for 12 hrs under nitrogen atmosphere. After the reaction was completed, the mixture was filtered and concentrated in vacuo. The residue was purified by column chromatography (EtOAc in PE = 10% ~ 25%) to afford 4-[5-bromo-6-[(15)- l-methoxyethyl]-3-pyridyl]morpholine (compound 12, 2100.0 mg) as colorless oil. MS calc’d

301.1 (MH ⁺ ), measured 301.1 (MH ⁺ ).

Step 2: Preparation of 4-[6-[(lS)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-3-pyridyl]morpholine (compound 13)

To a mixture of 4-[5-bromo-6-[(15)-l-methoxyethyl]-3-pyridyl]morpholine (compound 12,

2.1 g, 6.97 mmol) and bis(pinacolato)diboron (2.1 g, 8.37 mmol) in 1,4-dioxane (25 mL) were added KOAc (1.7 g, 17.43 mmol) and Pd(dppf)C12 (0.2 g, 0.35 mmol). The mixture was stirred at 90 °C for 12 hrs under nitrogen atmosphere. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by column chromatography (EtOAc in PE = 10% ~ 25%) to afford 4-[6-[(15)-l-methoxyethyl]-5- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3-pyridyl]morp holine (compound 13, 1100.0 mg) as a brown solid. MS calc’d 349.2 (MH ⁺ ), measured 349.2 (MH ⁺ )

Step 3: Preparation of [3-[5-bromo-6-fluoro-2-[2-[(lS)-l-methoxyethyl]-5- morpholino-3-pyridyl]-lH-indol-3-yl]-2,2-dimethyl-propoxy]-t ert-butyl-diphenyl-silane (compound 14)

To a mixture of [3-(5-bromo-6-fhioro-2-iodo-l/Z-indol-3-yl)-2,2-dimethyl-pro poxy]-Z€rZ- butyl-diphenyl- silane (compound D6, 1.9 g, 2.94 mmol) and 4-[6-[(15)-l-methoxyethyl]-5- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3-pyridyl]morp holine (compound 13, 1.0 g, 2.94 mmol) in 1,4-dioxane (25 mL) and water (5 mL) were added potassium phosphate (1.8 g, 8.64 mmol) and Pd(dppf)Ch (0.22 g, 0.29 mmol). The mixture was stirred at 70 °C for 12 hrs under nitrogen atmosphere. After the reaction was completed, the reaction mixture was filtered and the filtrated was diluted with water (50 mL) and extracted with EtOAc (100 mL, three times). The combined organic phase was washed with brine (200 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography to afford [3-[5- bromo-6-fluoro-2-[2-[(lS)-l-methoxyethyl]-5-morpholino-3-pyr idyl]-l/Z-indol-3-yl]-2,2- dimethyl-propoxy]-?er?-butyl-diphenyl- silane (compound 14, 1.4 g) as yellow gum.MS calc’d 758.2 (MH ⁺ ), measured 758.2 (MH ⁺ ).

Step 4: Preparation of [3-[5-bromo-6-fluoro-(2Af)-2-[2-[(lS)-l-methoxyethyl]-5- morpholino-3-pyridyl]-l-(2,2,2-trifluoroethyl)indol-3-yl]-2, 2-dimethyl-propoxy]-tert-butyl- diphenyl-silane (compound 15)

To a solution of [3-[5-bromo-6-fluoro-2-[2-[(15)-l-methoxyethyl]-5-morpholino -3- pyridyl]-l/7-indol-3-yl]-2,2-dimethyl-propoxy]-terZ-butyl-di phenyl-silane (compound 14, 1.4 g, 1.89 mmol) and cesium carbonate (1.8 g, 5.67 mmol) in DMF (30 mL) was added 2,2,2- trifluoroethyl trifluoromethanesulfonate (4.4 g, 18.9 mmol) dropwise at 0 °C, and the mixture was stirred at 30 °C for 16 hrs. After the reaction was completed, EtOAc (200 mL) and water (200 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (100 mL, twice). The combined organic layer was washed with brine (100 mL, three times), dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by column chromatography (EtOAc in PE = 25% ~ 66%) to afford [3-[5-bromo-6-fluoro-(2A7)-2-[2- [(lS)-l-methoxyethyl]-5-morpholino-3-pyridyl]-l-(2,2,2-trifl uoroethyl)indol-3-yl]-2,2-dimethyl- propoxy]-Z€rZ-butyl-diphenyl-silane (compound 15, 800.0 mg, PEAK 1, faster eluted) as a brown solid. MS calc’d 840.1 (MH ⁺ ), measured 840.1 (MH ⁺ ).

Step 5: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(lS)-l-methoxyethyl]-5- morpholino-3-pyridyl]-l-(2,2,2-trifluoroethyl)indol-3-yl]-2, 2-dimethyl-propan-l-ol (compound 16)

To a solution of [3-[5-bromo-6-fluoro-(2A7)-2-[2-[(lS)-l-methoxyethyl]-5-morp holino-3- pyridyl]-l-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-pr opoxy]-tert-butyl-diphenyl-silane (compound 15, 0.8 g, 0.95 mmol) in DMF (10 mL) was added cesium fluoride (0.7 g, 4.93 mmol) and the mixture was stirred at 60 °C for 15 hrs. After the reaction was completed, the reaction mixture was cooled to room temperature. EtOAc (70 mL) and water (100 mL) were added and layers were separated. The aqueous phase was extracted with EtOAc (70 mL, twice). The combined organic layer was washed with brine (80 mL, four times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by column chromatography (EtOAc in PE = 10% ~ 66%) to afford 3-[5-bromo-6-fluoro-(2A7)-2-[2-[(15)-l- methoxyethyl]-5-morpholino-3-pyridyl]-l-(2,2,2-trifluoroethy l)indol-3-yl]-2,2-dimethyl-propan- l-ol (compound 16, 430.0 mg) as yellow gum. MS calc’d 602.1 (MH ⁺ ), measured 602.2 (MH ⁺ ) Step 6: Preparation of 3-[6-fluoro-(2Af)-2-[2-[(lS)-l-methoxyethyl]-5-morpholino-3- pyridyl]-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l-( 2,2,2-trifluoroethyl)indol-3-yl]- 2,2-dimethyl-propan-l-ol (compound 17)

To a solution of 3-[5-bromo-6-fluoro-(2A7)-2-[2-[(lS)-l-methoxyethyl]-5-morph olino-3- pyridyl]-l-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-pr opan-l-ol (compound 16, 430.0 mg, 0.71 mmol) and bis(pinacolato)diboron (199.3 mg, 0.79 mmol) in toluene (5 mL) was added potassium acetate (0.1 mL, 1.78 mmol) and Pd(dppf)Ch (52.2 mg, 0.07 mmol) and the mixture was degassed by bubbling nitrogen for 2 min and then the mixture was stirred at 90 °C for 15 hrs under nitrogen atmosphere. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE = 10% ~ 66%) to afford 3-[6-fluoro-(2A7)-2-[2-[(15)-l- methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl -l,3,2-dioxaborolan-2-yl)-l- (2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-l-ol (compound 17, 390.0 mg) as colorless gum. MS calc’d 650.3 (MH ⁺ ), measured 650.3 (MH ⁺ )

Step 7: Preparation of methyl (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6- fhioro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2Af)-2-[2-[(lS)-l- methoxyethyl]-5-morpholino- 3-pyridyl]-l-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]p ropanoyl]hexahydropyridazine-3- carboxylate(compound 18)

To a mixture of 3-[6-fluoro-(2A7)-2-[2-[(lS)-l-methoxyethyl]-5-morpholino-3- pyridyl]-5- (4, 4, 5, 5 -tetramethyl- 1,3, 2-dioxaboro lan-2-yl)-l-(2, 2, 2-trifluoroethyl)indo 1-3-yl] -2,2-dimethyl- propan-l-ol (compound 17, 390.0 mg, 0.6 mmol) and methyl (3S)-l-[(2S)-3-(4-bromothiazol-2- yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazin e-3-carboxylate (intermediate C, 291.6 mg, 0.61 mmol) in toluene (4 mL), 1,4-dioxane (1.3 mL) and water (1.3 mL) were added K3PO4 (320.06 mg, 1.51 mmol) and Pd(dtbpf)C12 (39.1 mg, 0.06 mmol). The mixture was stirred at 70 °C for 12 hrs under nitrogen atmosphere. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by column chromatography (EtOAc in PE = 10% ~ 75%) to afford (3S)-l-[(2S)-2-(tert- butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethy l-propyl)-(2A7)-2-[2-[(lS)-l- methoxyethyl]-5-morpholino-3-pyridyl]-l-(2,2,2-trifluoroethy l)indol-5-yl]thiazol-2- yl]propanoyl]hexahydropyridazine-3-carboxylate (compound 18, 200.0 mg) as a brown solid. MS calc’d 920.3 (MH ⁺ ), measured 920.3 (MH ⁺ ).

Step 8: Preparation of (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-( 3- hydroxy-2,2-dimethyl-propyl)-(2Af)-2-[2-[(lS)-l-methoxyethyl ]-5-morpholino-3-pyridyl]-l- (2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexa hydropyridazine-3-carboxylic add (compound 19)

To a solution of (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-( 3-hydroxy- 2,2-dimethyl-propyl)-(2A7)-2-[2-[(lS)-l-methoxyethyl]-5-morp holino-3-pyridyl]-l-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropy ridazine-3-carboxylate (compound 18, 170.0 mg, 0.18 mmol) in DCE (4 mL) was added trimethylstannanol (133.6 mg, 0.74 mmol). The mixture was stirred at 60 °C for 15 hrs. After the reaction was completed, the reaction mixture was cooled to room temperature. The reaction mixture was concentrated, then EtOAc (40 mL) and water (50 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (50 mL, twice). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under vacuum to afford (3S)- l-[(2S)-2-(/e/7- butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethy l-propyl)-(2A7)-2-[2-[(lS)-l- methoxyethyl]-5-morpholino-3-pyridyl]-l-(2,2,2-trifluoroethy l)indol-5-yl]thiazol-2- yl]propanoyl]hexahydropyridazine-3-carboxylic acid (compound 19, 160.0 mg) as a yellow solid. MS calc’d 906.3 (MH ⁺ ), measured 906.3 (MH ⁺ ).

Step 9: Preparation of tert-butyl A^-[(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-di oxo-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamate (compound 110)

To a solution of (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-( 3-hydroxy- 2,2-dimethyl-propyl)-(2A7)-2-[2-[(lS)-l-methoxyethyl]-5-morp holino-3-pyridyl]-l-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropy ridazine-3-carboxylic acid (compound 19, 160.0 mg, 0.18 mmol) in DCM (15 mL) was added EDCI (507.8 mg, 2.65 mmol) and DIEA (0.6 mL, 3.53 mmol), then followed by addition of HOBt (59.6 mg, 0.44 mmol) at 0 °C, and then the mixture was stirred at 20 °C for 15 hrs. After the reaction was completed, EtOAc (50 mL) and water (80 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (50 mL, twice). The combined organic layer was washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by column chromatography (EtOAc in PE = 25% -66%) to afford tertbutyl A-[(7S,13S)-24-fhioro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-mo rpholino-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamate (compound 110, 130.0 mg) as an off-white solid. MS calc’d 888.3 (MH ⁺ ), measured 888.3 (MH ⁺ )

Step 10: Preparation of (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(lS)-l- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2 ,2-trifluoroethyl)-15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ^2,5 .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaene-8, 14-dione (Intermediate I)

To a solution of tert-butyl A-[(75, 13S)-24-fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5- morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen- 7 -yl] carbamate (compound 110, 130.0 mg, 0.15 mmol) in DCM (1.5 mL) was added TFA (0.6 mL, 8.27 mmol) and the mixture was stirred at 20 °C for 1 h. After the reaction was completed, sat. NaHCCh solution (40 mL) was added and the mixture was extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford (7S,13S)-7-amino-24-fluoro- (20A7)-20-[2-[(15)-l-methoxyethyl]-5-morpholino-3-pyridyl]-1 7,17-dimethyl-21-(2,2,2-

25 913 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate I, 105.0 mg) as a yellow solid. MS calc’d 788.3 (MH ⁺ ), measured 788.3 (MH ⁺ ).

Intermediate J

(7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyeth yl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-21- (2,2,2-trifluoroethyl)-15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ^2,5 .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaene-8, 14-dione The title compound was prepared in analogy to the preparation of Intermediate I by using l-(2,2,2-trifluoroethyl)piperazine instead of morpholine.

Intermediate K

(8S,14S)-8-amino-26-fluoro-4-hydroxy-(21M)-21-[2-[(lS)-l- methoxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-18,18-dimethyl-22-(2,2,2-tr ifluoroethyl)-16-oxa-10,22,28- triazapentacyclo[18.5.2.1 ² ’ ⁶ .l ¹⁰ ’ ¹⁴ .0 ²³ ’ ²⁷ ]nonacosa-l(26), 2, 4, 6(29), 20, 23(27), 24-heptaene-9, 15- dione

The title compound was prepared in analogy to the preparation of Intermediate G by using benzyl 4-[(5Af)-5-[5-bromo-4-fluoro-3-(3-hydroxy-2,2-dimethyl-propy l)-l-(2,2,2- trifluoroethyl)indol-2-yl]-6-[(lS)-l-methoxyethyl]-3-pyridyl ]piperazine-l-carboxylate (compound K8) instead of benzyl 4-[(5Af)-5-[5-bromo-l-ethyl-6-fluoro-3-(3-hydroxy-2,2- dimethyl-propyl)indol-2-yl]-6-[(lS)-l-methoxyethyl]-3-pyridy l]piperazine-l-carboxylate (compound D8). benzyl 4-[(5Af)-5-[5-bromo-4-fluoro-3-(3-hydroxy-2,2-dimethyl-propy l)-l-(2,2,2- trifluoroethyl)indol-2-yl]-6-[(lS)-l-methoxyethyl]-3-pyridyl ]piperazine-l-carboxylate (compound K8) was prepared in analogy to the preparation of benzyl 4-[(5Af)-5-[5-bromo-l- ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)indol-2-yl]- 6-[(15)-l-methoxyethyl]-3- pyridyl]piperazine-l -carboxylate (compound D8) using 5-bromo-4-fluoro-l//-indole instead of 5-bromo-6-fluoro- l /7- indole (compound D2).

Intermediate L

(7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-m ethoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin- 1-yl] -3-pyridyl] - 17,17-dimethyl- 15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14- dione

The title compound was prepared in analogy to the preparation of Intermediate I by using l-(2,2,2-trifluoroethyl)piperazine and iodoethane instead of morpholine and 2,2,2-trifluoroethyl trifluoromethanesulfonate.

Intermediate M

(7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l- methoxyethyl]-5-morpholino-3- pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapent acyclo-

[17.5.2.1 ^2,5 .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione

M7 intermediate M

Step 1: Preparation of [3-[5-bromo-l-ethyl-6-fluoro-2-[2-[(lS)-l-methoxyethyl]-5- inorpholino-3-pyri(lyl|in(lol-3-yl|-2.2-dimethyl-propoxy ]-/fr/-butyl-(liphenyl-silane (compound Ml)

To a solution of [3-[5-bromo-6-fluoro-2-[2-[(lS)-l-methoxyethyl]-5-morpholino -3- pyridyl]-l//-indol-3-yl]-2,2-dimethyl-propoxy]-terZ-butyl-di phenyl-silane (compound 14, 15 g, 19.77 mmol) in DMF (300 mL) was added CS2CO3 (19.3 g, 59.3 mmol) and iodoethane (6.16 g, 39.53 mmol) at 0 °C. After being stirred at 20 °C for 16 hrs, the reaction mixture was poured into water (200 mL), and extracted with EtOAc (200 mL, three times). The combined organic layer was washed with brine (10 mL, three times), dried over Na2SO4, filtered and concentrated under vacuum to give a residue. The residue was purified by column chromatography to afford [3-[5- bromo-l-ethyl-6-fluoro-2-[2-[(lS)-l-methoxyethyl]-5-morpholi no-3-pyridyl]indol-3-yl]-2,2- dimethyl-propoxy]-terZ-butyl-diphenyl- silane (compound Ml, 14.7 g) as yellow oil. MS calc’d 786.3 (MH ⁺ ), measured 786.4 (MH ⁺ ).

Step 2: Preparation of 3-[5-bromo-l-ethyl-6-fluoro-(2Af)-2-[2-[(lS)-l-methoxyethyl] - 5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propan-l-ol (compound M2) and 3-[5- bromo-l-ethyl-6-fluoro-(2P)-2-[2-[(lS)-l-methoxyethyl]-5-mor pholino-3-pyridyl]indol-3- yl]-2,2-dimethyl-propan-l-ol (compound M3)

To a solution of [3-[5-bromo-l-ethyl-6-fluoro-2-[2-[(lS)-l-methoxyethyl]-5-mo rpholino- 3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphe nyl-silane (compound Ml, 14.7 g, 18.68 mmol) in DMF (160 mL) was added cesium fluoride (14.2 g, 93.41 mmol). The mixture was stirred at 60 °C for 48 hrs. After being cooled to room temperature, the reaction mixture were added with EtOAc (300 mL) and water (300 mL) and the layers were separated. The aqueous phase was extracted with EtOAc (200 mL, three times). The combined organic layer was washed with brine (200 mL, four times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by column chromatography to afford 3-[5- bromo- l-ethyl-6-fhioro-(2M)-2-[2-[( IS)- l-methoxyethyl]-5-morpholino-3-pyridyl]indo 1-3-yl]- 2,2-dimethyl-propan-l-ol (compound M2, 6 g, faster eluted) as colorless foam and 3-[5-bromo- l-ethyl-6-fluoro-(2P)-2-[2-[(lS)-l-methoxyethyl]-5-morpholin o-3-pyridyl]indol-3-yl]-2,2- dimethyl-propan-l-ol (compound M3, 4.5 g, slower eluted) as colorless foam. Compound M2: MS calc’d 548.2 (MH ⁺ ), measured 548.2 (MH ⁺ ). ^J H NMR (400MHz, Methanol-^) d = 8.41 (d, J = 2.4 Hz, 1H), 7.92 (d, J = 6.8 Hz, 1H), 7.37 - 7.33 (m, 2H), 4.58 (s, 1H), 4.05 - 3.98 (m, 2H), 3.87-3.82 (m, 5H), 3.27 - 3.23 (m, 4H), 3.15 - 3.13 (m, 1H), 3.00 (s, 3H), 2.75-2.71 (m, 1H), 2.24 - 2.22 (m, 1H), 1.42 (d, J = 6.4 Hz, 3H), 1.22 (t, J = 7.2 Hz, 3H), 0.76 (s, 3H), 0.76 (s, 3H).

X-ray crystallographic analysis of compound M2

Absolute configuration structure of compound M2 was confirmed by X-ray crystallographic analysis of its single crystal. (Figure 3). Step 3: Preparation of 3-[l-ethyl-6-fluoro-(2Af)-2-[2-[(lS)-l-methoxyethyl]-5- morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-l,3,2-dioxaboro lan-2-yl)indol-3-yl]-2,2- dimethyl-propan-l-ol (compound M4)

To a solution of 3-[5-bromo-l-ethyl-6-fluoro-(2Af)-2-[2-[(lS)-l-methoxyethyl] -5- morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propan-l-ol (compound M2, 6 g, 10.94 mmol), bis(pinacolato)diboron (4.2 g, 16.41 mmol) in toluene (60 mL) was added potassium acetate (2.7 g, 27.35 mmol) and Pd(dppf)Ch (0.8 g, 1.09 mmol). The reaction mixture was degassed by bubbling nitrogen for 5 min then stirred at 90 °C for 15 hrs. After being cooled to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography to afford 3-[l-ethyl-6-fluoro-(2Af)- 2-[2-[(15)-l-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5, 5-tetramethyl-l,3,2-dioxaborolan-

2-yl)indol-3-yl]-2,2-dimethyl-propan-l-ol (compound M4, 4.5 g) as colorless gum. MS calc’d 596.4 (MH ⁺ ), measured 596.4 (MH ⁺ ).

Step 4: Preparation of methyl (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[l- ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2Af)-2-[2- [(lS)-l-methoxyethyl]-5- morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexah ydropyridazine-3- carboxylate (compound M5)

To a mixture of 3-[l-ethyl-6-fluoro-(2A7)-2-[2-[(lS)-l-methoxyethyl]-5-morph olino-3- pyridyl]-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)indo l-3-yl]-2,2-dimethyl-propan-l-ol (compound M4, 4.5 g, 7.56 mmol) and methyl (3S)-l-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ate (intermediate C, 3.6 g, 7.56 mmol) in toluene (45 mL), 1,4-dioxane (15 mL) and water (15 mL) were added K3PO4 (4.0 g, 18.89 mmol) and Pd(dtbpf)C12 (492.5 mg, 0.75 mmol). The mixture was stirred at 70 °C for 12 hrs under nitrogen atmosphere. After being cooled to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography to afford methyl (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[l- ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2A7)-2-[2- [(15)-l-methoxyethyl]-5- morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexah ydropyridazine-3-carboxylate (compound M5, 3.8 g) as colorless gum. MS calc’d 866.4 (MH ⁺ ), measured 866.4 (MH ⁺ ).

Step 5: Preparation of (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[l-ethyl-6- fhioro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2Af)-2-[2-[(lS)-l- methoxyethyl]-5-morpholino-

3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyri dazine-3-carboxylic acid (compound M6) To a mixture of methyl (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[l-ethyl-6-fl uoro-3- (3-hydroxy-2,2-dimethyl-propyl)-(2A7)-2-[2-[(15)-l-methoxyet hyl]-5-morpholino-3- pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazin e-3-carboxylate (compound M5, 3.8 g, 4.39 mmol) in DCE (76 mL) was added MesSnOH (3.2 g, 17.55 mmol). The mixture was stirred at 60 °C for 48 hrs. The reaction mixture was concentrated under vacuum to give a residue. EtOAc (200 mL) and water (100 mL) were added to the residue and the layers were separated. The aqueous phase was extracted with EtOAc (150 mL, twice). The combined organic layer was washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under vacuum to afford (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[l-ethyl-6-fl uoro-3-(3- hydroxy-2,2-dimethyl-propyl)-(2A7)-2-[2-[(lS)-l-methoxyethyl ]-5-morpholino-3-pyridyl]indol- 5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (compound M6, 3.7 g) as a brown solid. MS calc’d 852.4 (MH ⁺ ), measured 852.4 (MH ⁺ ).

Step 6: Preparation of tert-butyl A-[(7S, 13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-di oxo-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]carbamate (compound M7)

To a mixture of (3S)-l-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[l-ethyl-6-fl uoro-3-(3- hydroxy-2,2-dimethyl-propyl)-(2A7)-2-[2-[(lS)-l-methoxyethyl ]-5-morpholino-3-pyridyl]indol- 5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (compound M6, 2.5 g, 2.93 mmol) in DCM (250 mL) were added DIEA (7.58 mL, 58.68 mmol), EDCI (8.4 g, 44.01 mmol) and HOBt (991.2 mg, 0.91 mmol) at 0 °C. After being stirred at 20 °C for 12 hrs, the reaction mixture was poured into water (100 mL) and extracted with EtOAc (100 mL, three times). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under vacuum to give a residue which was purified by column chromatography to afford tert-butyl A-[(7S,13S)-21-ethyl-24-fluoro-(20A7)-20-[2-[(15')-l-methoxy ethyl]-5- morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thi a-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamate (compound M7, 1.2 g) as a yellow oil. MS calc’d 834.4 (MH ⁺ ), measured 834.4 (MH ⁺ ).

Step 7: Preparation of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa- 4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14- dione (Intermediate M) To a solution of ferLbutyl A-[(75,135)-21-ethyl-24-fluoro-(20Af)-20-[2-[(15)-l- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-di oxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamate (compound M7, 1.2 g, 1.44 mmol) in DCM (12 mL) was added TFA (6.0 mL). The mixture was stirred at 20 °C for 3 hrs. After the reaction was completed, the reaction mixture was concentrated under vacuum to give a residue. Sat. NaHCCL aq. (60 mL) was added and the mixture was extracted with EtOAc (80 mL, three times). The combined organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford (7S,13S)-7-amino-21-ethyl-24-fhioro-(20Af)-20-[2-[(lS)-l-met hoxyethyl]-5- morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,2 8- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate M, 1 g) as a yellow solid. MS calc’d 734.3 (MH ⁺ ), measured 734.3 (MH ⁺ ).

Intermediate N

(7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-m ethoxyethyl]-3-pyridyl]-17,17-

25 9 13 2226 dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione

The title compound was prepared in analogy to the preparation of Intermediate I by using by using 2-[(15)-l-methoxyethyl]-3-(4,4,5,5-tetramethyl-l,3,2-dioxabo rolan-2-yl)pyridine (compound Nl) instead of 4-[6-[(15)-l-methoxyethyl]-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-3-pyridyl]morpholine (compound 13).

The compound Nl was prepared in analogy to the preparation of Intermediate A by using 3-bromo-2-[(15)-l-methoxyethyl]pyridine (compound Al) instead of benzyl 4-[5-bromo-6- [( lS)- l-mcthoxycthyl]-3-pyridyl]pipcrazinc- l -carboxylate (Intermediate A5). Example 1

(3S)-l-[(2/f)-2-chloro-2-fluoro-acetyl]-A^-[(lS)-l-[[(7S, 13S)-25-fluoro-(20Af)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-2V-methyl-pyrroli dine-

3-carboxamide

The compound was prepared according to the following scheme:

EXAMPLE 1

Step 1: Preparation of te^-butyl ^-[(15)-l-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(15)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-2V-methyl-carbama te

(compound IB).

To a solution of (7S,13S)-7-amino-25-fluoro-(20Af)-20-[2-[(15')-l-methoxyethy l]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate E, 150.0 mg, 0.19 mmol) in DMF (1.5 mL) was added a solution of BOC-N-ME-VAL-OH (compound 1A, 75.0 mg, 0.32 mmol), DIEA (0.1 mL, 0.58 mmol) and HATU (120.0 mg, 0.32 mmol) in DMF (1.5 mL) at 0 °C and the mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was poured into water (30 mL), and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (30 mL, three times), dried over Na2SO4, filtered and concentrated under vacuum to give a residue which was purified by prep-HPLC. The eluent was concentrated in vacuo to give tertbutyl A-[(15)-l-[[(7S,13S)-25-fhioro-(20M)-20-[2-[(lS)-l-methoxyet hyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluo roethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-A-methyl-carbamate (compound IB, 150.0 mg) as a yellow solid. MS calc’d 1014.5 (MH ⁺ ), measured 1014.5 (MH ⁺ ).

Step 2: Preparation of (2S)-jV-[(7S,13S)-25-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl ]- 5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-di oxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-2-(methylamino)butanamide (compound 1C).

To a solution of tert-butyl A-[(15)-l-[[(75,135)-25-fluoro-(20Af)-20-[2-[(15)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 913 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-A-methyl-carbamate (compound IB, 100.0 mg, 0.1 mmol) in DCM (1 mL) was added TFA (1.0 mL) and the mixture was stirred at 20 °C for 2 hrs. After the reaction was completed, the reaction mixture was concentrated under vacuum to give a residue. Sat. NaHCCh aqueous solution (20 mL) was added and the mixture was extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford (2S)-A-[(7S,13S)-25-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl] -5-(4- methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]-3-methyl-2-(methylamino)butanamide (compound 1C, 90.0 mg) as a yellow solid. MS calc’d 914.4 (MH ⁺ ), measured 914.4 (MH ⁺ ). Step 3: Preparation of tert-butyl (3S)-3-[[(lS)-l-[[(7S,13S)-25-fluoro-(20M)-20-[2- [(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-21- (2,2,2-trifhioroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine- l-carboxylate (compound IE).

To a solution of (25)-A-[(75,135)-25-fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5-(4- methylpiperazin- 1 -yl)-3 -pyridyl] - 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]-3-methyl-2-(methylamino)butanamide (compound 1C, 90.0 mg, 0.1 mmol) in DMF (1.5 mL) was added a solution of (S)-l-Boc-pyrrolidine-3-carboxylic acid (compound ID, 31.9 mg, 0.15 mmol), DIEA (0.1 mL, 0.33 mmol), HATU (57.2 mg, 0.15 mmol) at 0 °C and then the mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was poured into water (20 mL), and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered and concentrated under vacuum to give a residue which was purified by prep-HPLC to give tertbutyl (35)-3-[[(15)-l-[[(75,135)-25-fluoro-(20M)-20-[2-[(15)-l-met hoxyethyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl] carbamoyl] -2-methyl-propyl] -methyl-carbamoyl]pyrrolidine- 1 -carboxylate (compound IE, 40.0 mg) as a yellow solid. MS calc’d 1111.5 (MH ⁺ ), measured 1111.6 (MH ⁺ ).

Step 4: Preparation of (3S)-2V-[( lS)-l-[[(7S,13S)-25-fluoro-(20M)-20-[2-[( 1S)-1- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2- 25 9 13 2226 trifhioroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacydo[17 .5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbanioyl|-2-niethyl-propyl|-\-methyl-pyrrol idine- 3-carboxamide (compound IF).

To a solution of tert-butyl (3S)-3-[[(15)-l-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 9 13 22 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-methyl- carbamoyl]pyrrolidine-l -carboxylate (compound IE, 40.0 mg, 0.04 mmol) in DCM (1 mL) was added TFA (0.3 mL), and the mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated under vacuum to give a residue. Sat. NaHCCL aqueous solution (20 mL) was added and the mixture was extracted with EtOAc (15 mL, three times). The combined organic layer was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford (35 ^, )-A-[(lS)-l-[[(7S,135')-25- fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-A-methyl-pyrrolidine-3-carbox amide (compound IF, 25.0 mg) as a yellow solid. MS calc’d 1011. 4 (MH ⁺ ), measured 1011.4 (MH ⁺ ).

Step 5: Preparation of (3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,13S) -25- fhioro-(20V7)-20-|2-|( LS)- l-inethoxyethyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl|- 17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo- [17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]carba moyl]-2-methyl- propyl]-A-methyl-pyrrolidine-3-carboxamide (Example 1).

To a solution of (27?)-2-chloro-2-fluoro-acetic acid (23.0 mg, 0.2 mmol), (3S)-A-[(1S)-1- [[(7S,13S)-25-fhioro-(20M)-20-[2-[(15)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-A-methyl-pyrrolidine-3-carbox amide (compound IF, 25.0 mg, 0.02 mmol) and DIEA (0.1 mL, 0.77 mmol) in DMF (2 mL) was added T3P (130.0 mg, 0.2 mmol) at 0 °C, and the mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was poured into water (20 mL), and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (30 mL, three times), dried over Na2SO4, filtered and concentrated under vacuum to give a residue which was purified by prep- HPLC to afford (35)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-A-[(15)-l-[[(75,135) -25-fluoro-(20Af)-

20-[2-[(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-

21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraz apentacyclo-

[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]carba moyl]-2-methyl- propyl]-A-methyl-pyrrolidine-3-carboxamide (Example 1, 9.4 mg) as a white solid. MS calc’d 1105.4 (MH ⁺ ), measured 1105.8 (MH ⁺ ).' H NMR (400 MHz, METHANOL-^) 6 = 8.52 (d, J = 2.8 Hz, 1H), 7.59 - 7.41 (m, 4H), 7.01 - 6.70 (m, 1H), 5.97 - 5.85 (m, 1H), 5.22 - 5.04 (m, 1H), 4.68 - 4.63 (m, 1H), 4.43 - 4.35 (m, 1H), 4.11 - 3.91 (m, 3H), 3.81 - 3.48 (m, 13H), 3.44 - 3.39 (m, 1H), 3.28 - 3.24 (m, 1H), 3.23 - 3.11 (m, 6H), 3.05 - 2.97 (m, 4H), 2.92 - 2.80 (m, 1H), 2.66 - 2.55 (m, 1H), 2.39 - 1.97 (m, 4H), 1.71 - 1.60 (m, 1H), 1.46 (d, J = 6.4 Hz, 3H), 1.35 - 1.27 (m, 2H), 1.18 (t, J = 7.2 Hz, 3H), 0.98 - 0.79 (m, 9H), 0.67 - 0.57 (m, 2H) ppm.

Example 2

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,1 3S)-24-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-2V-methyl-pyrroli dine- 3-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l-methoxyethy l]-5-(4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-t hia-9,21,27,28-tetrazapentacyclo- [17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate D) instead of (7S, 13S)-7-ainino-25-fhioro-(20A4)-20-[2-[( lS)- l -mcthoxycthyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate E). Example 2 (39.0 mg) was obtained as a white solid. MS calc’d 1105.4 (MH ⁺ ), measured 1105.6 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.68 (d, J = 7.6 Hz, 1H), 8.57 - 8.49 (m, 1H), 7.79 - 7.66 (m, 1H), 7.56 - 7.43 (m, 2H), 6.95 - 6.72 (m, 1H), 5.83 - 5.61 (m, 1H), 5.30 - 5.10 (m, 1H), 4.96 - 4.88 (m, 3H), 4.82 (dd, J = 11.2, 3.6 Hz, 1H), 4.52 - 4.36 (m, 1H), 4.29 - 4.17 (m, 2H), 4.15 - 3.93 (m, 2H), 3.90 - 3.67 (m, 6H), 3.66 - 3.55 (m, 2H), 3.55 - 3.41 (m, 2H), 3.36 (d, J = 2.8 Hz, 3H), 3.34 - 3.32 (m, 1H), 3.30 - 3.23 (m, 2H), 3.19 - 3.14 (m, 1H), 3.14 - 3.09 (m, 3H), 3.03 - 2.96 (m, 3H), 2.89 - 2.74 (m, 1H), 2.63 - 2.50 (m, 1H), 2.45 - 2.21 (m, 3H), 2.20 - 1.91 (m, 2H), 1.90 - 1.75 (m, 1H), 1.72 - 1.57 (m, 1H), 1.45 (d, J = 6.4 Hz, 3H), 1.11 - 0.98 (m, 3H), 0.98 - 0.94 (m, 3H), 0.94 - 0.84 (m, 3H), 0.54 - 0.34 (m, 3H) ppm.

Example 3

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,1 3S) -(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17,24 -trimethyl-8,14-dioxo-21- (2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-A^-methyl-pyrrolidine-3-carbo xamide

The title compound was prepared in analogy to the preparation of Example 1 by using (75 ^, ,13S)-7-amino-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]- 17,17,24-trimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9 ,21,27,28-tetrazapentacyclo- [17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate F) instead of (7S, 13S)-7-ainino-25-fhioro-(20A4)-20-[2-[( lS)- l -mcthoxycthyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate E). Example 3 (4.7 mg) was obtained as a white solid. MS calc’d 1101.5 (MH ⁺ ), measured 1101.6 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.52 - 8.47 (m, 1H), 8.15 - 7.98 (m, 1H), 7.54 - 7.37 (m, 3H), 6.95 - 6.70 (m, 1H), 5.85 - 5.75 (m, 1H), 5.35 - 5.07 (m, 2H), 4.78 - 4.67 (m, 1H), 4.50 - 4.38 (m, 1H), 4.25 - 4.14 (m, 1H), 4.12 - 3.95 (m, 2H), 3.91 - 3.55 (m, 10H), 3.50 - 3.07 (m, 5H), 3.00 (s, 3H), 2.79 - 2.64 (m, 1H), 2.62 - 2.43 (m, 4H),

2.40 - 2.12 (m, 4H), 2.10 - 1.84 (m, 3H), 1.72 - 1.49 (m, 3H), 1.47 - 1.41 (m, 3H), 1.39 - 1.32 (m, 2H), 1.29 - 1.23 (m, 3H), 0.98 - 0.95 (m, 3H), 0.95 - 0.91 (m, 3H), 0.88 - 0.84 (m, 3H), 0.43 - 0.36 (m, 3H) ppm. Example 4 and Example 5

(2S)-2-[(5/?)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2, 7-diazaspiro[4.4]nonan-2-yl]-2V-

[(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5- (4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 4)

(Example 4) (2S)-2-[(5S)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7-di azaspiro[4.4]nonan-2-yl]-2V- [(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 5)

(Example 5)

The Example 4 was prepared according to the following scheme:

4B EXAMPLE 4 Step 1: Preparation of tert-butyl (5S)-2-[(lS)-l-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifhioroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-l-oxo-2,7- diazaspiro[4.4]nonane-7-carboxylate (compound 4A) To a solution of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4] nonan-2-yl]- 3-methyl-butanoic acid (compound 4f 2, 85.0 mg, 0.25 mmol) in DMF (3 mL) was added HATU (94.9 mg, 0.25 mmol), DIEA (0.1 mL, 0.37 mmol). After being stirred at 20 °C for 0.2 hour, (7S,13S)-7-amino-24-fhioro-(20M)-20-[2-[(15)-l-methoxyethyl] -5-(4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-t hia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate D, 100.0 mg, 0.12 mmol) was added. The reaction mixture was stirred at 20 °C for another 2 hrs. After the reaction was completed, the reaction mixture was purified directly by reversed phase flash column and the eluent was concentrated under vacuum to remove ACN. The aqueous was neutralized with NaHCCE and extracted with EtOAc (10 mL, three times). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under vacuum to afford tert-butyl (55 ^, )-2-[(lS)-l-[[(7S,135')-24-fluoro-(20Af)-20-[2- [(15)- l-methoxyethyl]-5-(4-methylpiperazin- 1 -yl) - 3 -pyridyl] - 17, 17-dimethyl-8, 14-dioxo-21-

25 9 13 22226 (2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapenta cyclo[17.5.2.1 ’ .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-l-oxo-2,7- diazaspiro[4.4]nonane-7-carboxylate (compound 4A, 120.0 mg) as yellow solid. MS calc’d 1123.6 (MH ⁺ ), measured 1123.6 (MH ⁺ ).

Step 2: Preparation of (2S)-jV-[(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl ]- 5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-di oxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-2-[(5/f)-l-oxo-2,7-diazaspiro[4.4]n onan-2- yl]butanamide (compound 4B)

To a solution of tert-butyl (55)-2-[(15)-l-[[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-l-oxo-2,7- diazaspiro[4.4]nonane-7-carboxylate (compound 4A, 120.0 mg, 0.11 mmol) in DCM (3 mL) was added TFA (1.0 mL, 12.98 mmol). The reaction mixture was stirred at 20 °C for 0.5 h. After the reaction was completed, the reaction mixture was concentrated under vacuum. The residue was diluted with EtOAc (10 mL) and neutralized with NaHCO? solution. The organic phase was separated and the aqueous phase was then extracted with EtOAc (10 mL, twice). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under vacuum to afford (2S)-A ^/ -[(7S, L3S)-24-fluoro-(20A7)-20-[2-[( lS)- l-mcthoxycthyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9, 21, 27, 28-tctrazapcntacyclo[ 17.5.2.1 ² \ l ⁹ ' ³ .0 ²² ' ²⁶ ]octacosa- 1 (25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]-3-methyl-2-[(5R)-l-oxo-2,7-diazaspiro[4.4]nonan -2-yl]butanamide (compound 4B, 120.0 mg) as yellow solid. MS calc’d 1023.5 (MH ⁺ ), measured 1023.5 (MH ⁺ ).

Step 3: Preparation of (2S)-2-[(5/f)-7-[(2/f)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-A^-[(7S,13S)-24-fluoro-(20Af)-20- [2-[(lS)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-2 1-(2,2,2-trifluoroethyl)-15- oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-butanamide (Example 4)

To a solution of (25)-A-[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-2 1-(2,2,2-trifluoroethyl)-15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]-3-methyl-2-[(5R)-l-oxo-2,7-diazaspiro[4.4]nonan -2-yl]butanamide (compound 4B, 110.0 mg, 0.11 mmol), (27?)-2-chloro-2-fluoro-acetic acid (55.0 mg, 0.49 mmol), DIEA (0.1 mL, 0.55 mmol) in DMF (2 mL) was added T3P (275.0 mg, 0.43 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was added with H2O (30 mL) and then extracted with EA (10 mL, twice). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated under vacuum to give the residue. The residue was purified by prep-HPLC to afford (2S)-2-[(5R)-7-[(2R)-2-chloro-2- fluoro-acetyl]-l-oxo-2,7-diazaspiro[4.4]nonan-2-yl]-A-[(7S,1 3S)-24-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 913 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-butanamide (Example 4, 54.4 mg) as a yellow solid. MS calc’d 1117.4 (MH ⁺ ), measured 1117.4 (MH ⁺ ). 1H NMR (400 MHz, METHANOL-^) 8: 8.68 (d, J = 7.2 Hz, 1 H), 8.50 (d, J = 2.8 Hz, 1 H), 7.82 - 7.69 (m, 1 H), 7.52 - 7.45 (m, 2 H), 6.95 - 6.72 (m, 1 H), 5.72 - 5.65 (m, 1 H), 5.21 - 5.14 (m, 1 H), 4.46 - 4.38 (m, 1 H), 4.37 - 4.31 (m, 1 H), 4.27 - 4.20 (m, 2 H), 4.16 - 3.97 (m, 2 H), 3.97- 3.88 (m, 1 H), 3.88 - 3.72 (m, 4 H), 3.72 - 3.54 (m, 6 H), 3.53 - 3.44 (m, 3 H), 3.35 (d, 7=4.2 Hz, 2 H), 3.19 - 3.13 (m, 2 H), 3.00 (s, 3 H), 2.86 - 2.78 (m, 1 H), 2.62 - 2.51 (m, 1 H), 2.34 - 2.19 (m, 3 H), 2.18 - 2.07 (m, 3 H), 2.06 - 1.93 (m, 2 H), 1.89 - 1.77 (m, 1 H), 1.69 - 1.60 (m, 1 H), 1.45 (d, 7=6.0 Hz, 3 H), 1.37 - 1.26 (m, 2 H), 1.03 - 0.96 (m, 6 H), 0.93 - 0.89 (m, 3 H), 0.46 - 0.40 (m, 3 H) ppm.

Example 5 was prepared in analogy to the preparation of Example 4 by using (2S)-2-[(5/?J- 7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4]nonan-2-yl]-3 -methyl-butanoic acid (compound 4f 1) instead of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4] nonan-2-yl]-3- methyl- butanoic acid (compound 4f 2). Example 5 (44 mg) was obtained as a white solid. MS calc’d 1117.4 (MH ⁺ ), measured 1117.4 (MH ⁺ ). ' HNMR (400 MHz, METHANOL-^) 6 = 8.74 - 8.68 (m, 1H), 8.56 - 8.50 (m, 1H), 7.76 - 7.69 (m, 1H), 7.62 - 7.56 (m, 1H), 7.54 - 7.47 (m, 1H), 6.96 - 6.73 (m, 1H), 5.75 - 5.64 (m, 1H), 5.20 (dd, J = 8.4, 16.4 Hz, 1H), 4.46 - 4.40 (m, 1H), 4.35 (dd, J = 4.8, 11.1 Hz, 1H), 4.31 - 4.19 (m, 2H), 4.18 - 4.02 (m, 2H), 3.95 (dd, J = 4.9, 10.1 Hz, 1H), 3.85 - 3.72 (m, 4H), 3.71 - 3.61 (m, 3H), 3.56 - 3.42 (m, 4H), 3.38 (s, 4H), 3.27 (dd, J = 5.1, 10.0 Hz, 2H), 3.21 - 3.14 (m, 1H), 3.21 - 3.12 (m, 1H), 3.01 (s, 3H), 2.90 - 2.79 (m, 1H), 2.60 (d, J = 14.5 Hz, 1H), 2.33 - 2.20 (m, 3H), 2.17 - 2.05 (m, 3H), 2.02 - 1.92 (m, 1H), 1.90 - 1.78 (m, 1H), 1.76 - 1.60 (m, 1H), 1.48 (d, J = 6.1 Hz, 3H), 1.37 - 1.30 (m, 1H), 1.06 - 0.99 (m, 6H), 0.95 - 0.87 (m, 3H), 0.47 (s, 3H) ppm. The compound 4f 1 and 4f 2 were prepared according to the following scheme:

Step 1: Preparation of l-O('/7-butyl) 3-methyl 3-allylpyrrolidine-l,3-dicarboxylate (compound 4b). To a solution of 1 -(tert-butyl) 3-methyl pyrrolidine- 1,3-dicarboxylate (compound 4a, 5.0 g, 21.8 mmol) in THF (60 mL) was added LDA (12 mL, 24 mmol) dropwise at -70 °C under nitrogen atmosphere. After being stirred for 0.5 h, allyl bromide (2.9 g, 23.99 mmol) was added slowly. After the reaction was completed, the reaction mixture was poured into saturated NH4CI solution (100 mL) and extracted with EtOAc (70 mL, twice). The combined organic layer was washed with brine (70 mL ), dried over Na2SO4, filtered and concentrated under vacuum to give a residue, which was purified by silica gel column to afford 1 -(tert-butyl) 3-methyl 3- allylpyrrolidine- 1,3-dicarboxylate (compound 4b, 2.91 g) as colorless oil.

Step 2: Preparation of 1 -(tert-butyl) 3-methyl 3-(2-oxoethyl)pyrrolidine-l,3- dicarboxylate (compound 4c).

To the mixture of 1 -(tert-butyl) 3-methyl 3-allylpyrrolidine-l,3-dicarboxylate (compound 4b, 2.1 g, 7.8 mmol) in 1,4-dioxane (60 mL) and water (6 mL) was added 2,6-Lutidine (1.8 mL, 15.6 mmol) and K2OSO4 (0.1 g, 0.39 mmol) in one portion at 0 °C. After being stirred at 0 °C for 15 min, sodium metaperiodate (6.6 g, 31.19 mmol) was added portion-wise at 0 °C. The resulting mixture was warmed to 20 °C and stirred for another 6 hrs. After the reaction was completed, it was quenched with saturated aqueous solution (100 mL) and the reaction mixture was extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (50 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to afford 1 -(tert-butyl) 3-methyl 3-(2-oxoethyl)pyrrolidine-l,3-dicarboxylate (compound 4c, 2.1 g) as yellow oil, which was used in the next step directly.

Step 3: Preparation of 1 -(tert-butyl) 3-methyl 3-(2-(((S)-l-(benzyloxy)-3-methyl-l- oxobutan-2-yl)amino)ethyl)pyrrolidine-l,3-dicarboxylate (compound 4d).

To the mixture of 1 -(tert-butyl) 3-methyl 3-(2-oxoethyl)pyrrolidine-l,3-dicarboxylate (compound 4c, 2.1 g, 7.74 mmol) and benzyl (2S)-2-amino-3-methyl-butanoate (1.6 g, 7.74 mmol) in methanol (20 mL) was added zinc chloride (1.05 g, 7.74 mmol) in one portion at 0 °C. After being stirred at 0 °C for 1 h, to the mixture was added sodium cyanoborohydride (0.97 g, 15.48 mmol) at 0 °C. The resulting mixture was stirred at 0 °C for another 2 hrs. After the reaction was completed, the mixture was poured into saturated NH4CI aqueous solution (40 mL) at 0 °C and extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (30 mL, four times), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by silica gel column to afford 1 -(tert-butyl) 3-methyl 3-(2-(((S)-l-(benzyloxy)-3-methyl-l-oxobutan-2-yl)amino)ethy l)pyrrolidine-l,3- dicarboxylate (compound 4d, 2.2 g) as yellow oil. MS calc’d 463.3 (MH ⁺ ), measured 463.2 (MH ⁺ ).

Step 4: Preparation of tert-butyl 7-((S)-l-(benzyloxy)-3-methyl-l-oxobutan-2-yl)-6- oxo-2, 7-diazaspiro[4.4]nonane-2-carboxylate (compound 4e 1& 4e 2).

To the mixture of 1 -(tert-butyl) 3-methyl 3-(2-(((S)-l-(benzyloxy)-3-methyl-l-oxobutan-2- yl)amino)ethyl)pyrrolidine-l,3-dicarboxylate (compound 4d, 2.1 g, 4.54 mmol) in toluene (20 mL) was added DIEA (7.9 mL, 45.4 mmol) and DMAP (0.6 g, 4.54 mmol) in one portion. The mixture was heated to 80 °C and stirred for 16 hrs. After the reaction was completed, the mixture was poured into water (30 mL) and extracted with EtOAc (30 mL, three times). The combined organic layer was washed with brine (30 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to give a residue, which was purified by reversed phase flash column and the eluent was concentrated under vacuum. The residue was further separated by prep-SFC to afford tert-butyl 7-((S)-l-(benzyloxy)-3-methyl-l-oxobutan-2-yl)-6-oxo-2,7- diazaspiro[4.4]nonane-2-carboxylate (compound 4e 1, faster eluted, 521 mg, compound 4e 2, slower eluted, 525 mg). MS calc’d 453.3 (MNa ⁺ ), measured 453.2 (MNa ⁺ ).

Step 5: Preparation of (2.S )-2-|(5/?)-7-/er/-butoxycarbonyl-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-3-methyl-butanoic acid (compound 4f 1).

To a solution of tert-butyl (7?)-7-((S)-l-(benzyloxy)-3-methyl-l-oxobutan-2-yl)-6-oxo-2, 7- diazaspiro[4.4]nonane-2-carboxylate (compound 4e 1, 120 mg, 0.28mmol) in toluene (2 mL) was added wet palladium (12mg, 10% wt. on activated carbon). The mixture was degassed and purged with hydrogen for 3 times. The reaction mixture was heated to 35 °C and stirred for 3 hrs under hydrogen atmosphere. After the reaction was completed, the solution was filtered and the filtrate was concentrated in vacuo to give (2S)-2-[(57?)-7-tert-butoxycarbonyl-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-3-methyl-butanoic acid (compound 4f 1, 94 mg) as a white solid. MS calc’d 363.2 (MNa ⁺ ), measured 363.1 (MNa ⁺ ).

X-ray crystallographic analysis of compound 4f 1

Absolute configuration structure of compound 4f 1 was confirmed by X-ray crystallographic analysis of its single crystal. (Figure 1)

Step 6: Preparation of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-3-methyl-butanoic acid (compound 4f 2).

To a solution of tert-butyl (S)-7-((S)-l-(benzyloxy)-3-methyl-l-oxobutan-2-yl)-6-oxo-2,7 - diazaspiro[4.4]nonane-2-carboxylate (compound 4e 2, 120 mg, 0.28 mmol) in toluene (2 mL) was added wet palladium (12 mg, 10% wt. on activated carbon). The mixture was degassed and purged with hydrogen for 3 times. The mixture was heated to 35 °C and stirred for 3 h under hydrogen atmosphere. After the reaction was completed, the solution was filtered and the filtrate was concentrated in vacuo to afford (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-3-methyl-butanoic acid (compound 4f 2, 79 mg) as a white solid. MS calc’d 363.2 (MNa ⁺ ), measured 363.1 (MNa ⁺ ).

X-ray crystallographic analysis of compound 4f 2

Absolute configuration structure of compound 4f 2 was confirmed by X-ray crystallographic analysis of its single crystal. (Figure 2)

Example 6 (2.S)-2-|9-|(2/?)-2-chloro-2-fluoro-acetyl|-l -oxo-2, 9-diazaspiro|5.5]undecan-2-yl|-V- [(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The Example 6 was prepared according to the following scheme:

Step 1: Preparation of tert-butyl 2-[(lS)-l-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2- 25 9 13 2226 trifhioroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-l-oxo-2,9- diazaspiro[5.5]undecane-9-carboxylate (compound 6K)

To a solution of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2-yl)-3- methyl-butanoic acid (compound 6J, 138.0 mg, 0.37 mmol) in DMF (2 mL) was added DIEA (0.2 mL, 0.94 mmol), HATU (106.8 mg, 0.28 mmol). After being stirred at 20 °C for 10 min, the reaction mixture was added with a solution of (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)-

2 5 9 13 22 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D, 150.0 mg, 0.19 mmol) in DMF (2 mL) and the reaction mixture was stirred at 20 °C for 50 min. After the reaction was completed, the reaction mixture was added into water (20 mL) and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (20 mL, three times), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by reversed phase flash column to afford tert-butyl 2-[(15 ^, )-l-[[(7S,13S)-24-fluoro- (20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl )-3-pyridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-l-oxo-2,9-diazaspiro[5.5]unde cane-9-carboxylate (compound 6K, 125.0 mg) as a yellow solid. MS calc’d 1151.6 (MH ⁺ ), measured 1151.8 (MH ⁺ ).

Step 2: Preparation of (2S)-jV-[(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethy l]- 5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-di oxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-2-(l-oxo-2,9-diazaspiro[5.5]undecan -2- yl)butanamide (compound 6L)

To a solution of tert-butyl 2-[(15)-l-[[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 913 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-p ropyl]-l-oxo-2,9- diazaspiro[5.5]undecane-9-carboxylate (compound 6K, 125.0 mg, 0.11 mmol) in DCM (1.5 mL) was added TFA (0.6 mL, 7.73 mmol). The mixture was stirred at 20 °C for 0.5 h. After the reaction was completed, the reaction mixture was added into sat. NaHCCL aq. (20 mL) and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (20 mL, three times), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford (2S)-A ^/ -[(7S, L3S)-24-fhioro-(20A7)-20-[2-[( lS)- l -mcthoxycthyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]-3-methyl-2-(l-oxo-2,9-diazaspiro[5.5]undecan-2- yl)butanamide (compound 6L, 110.0 mg) as a yellow solid. MS calc’d 1051.5 (MH ⁺ ), measured 1051.7 (MH ⁺ ).

Step 3: Preparation of (2S)-2-[9-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9- diazaspiro|5.5]undecan-2-yl]-V-|(7.S, L3.S)-24-fhioro-(20V7)-20-|2-|( LS)-l-methoxyethyl]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-diox o-21-(2,2,2-trifluoroethyl)-15- oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-butanamide (Example 6) To a solution of (25)-A-[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5-(4- methylpiperazin- 1 -yl)-3 -pyridyl] - 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]-3-methyl-2-(l-oxo-2,9-diazaspiro[5.5]undecan-2- yl)butanamide (compound 6L, 110.0 mg, 0.1 mmol), (7?)-2-chloro-2-fluoroacetic acid (58.8 mg, 0.52 mmol) in DMF (3 mL) was added DIEA (0.1 mL, 0.52 mmol) and T3P (99.9 mg, 0.16 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was poured into water (40 mL) and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (20 mL, three times), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by prep-HPLC to afford (25)- 2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diazaspiro[5. 5]undecan-2-yl]-A-[(75,135)-24- fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ ,0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19 ,22(26), 23 -hexaen-7 -yl]-3- methyl-butanamide (Example 6, 17.7 mg) as a white solid. MS calc’d 1145.5 (MH ⁺ ), measured 1145.3 (MH ⁺ ). ^J H NMR (400MHz, Methanol-^) d = 8.69 (d, J = 6.4 Hz, 1H), 8.50 (d, J = 2.8 Hz, 1H), 7.74 - 7.70 (m, 1H), 7.53 - 7.45 (m, 2H), 7.09 - 6.91 (m, 1H), 5.70 - 5.63 (m, 1H), 5.22 - 5.13 (m, 1H), 4.97 - 4.89 (m, 3H), 4.83 - 4.77 (m, 1H), 4.44 - 4.36 (m, 1H), 4.27 - 4.20 (m, 2H), 4.17 - 3.89 (m, 4H), 3.80 - 3.69 (m, 3H), 3.65 - 3.59 (m, 1H), 3.52 - 3.41 (m, 4H), 3.36 (s, 5H), 3.24 - 3.11 (m, 3H), 2.99 (s, 3H), 2.87 - 2.78 (m, 1H), 2.61 - 2.54 (m, 1H), 2.31 - 2.08 (m, 4H), 1.97 - 1.80 (m, 6H), 1.69 - 1.59 (m, 3H), 1.45 (d, J = 6.0 Hz, 3H), 1.02 - 0.94 (m, 6H), 0.87 (d, J = 6.4 Hz, 3H), 0.44 (s, 3H) ppm.

The compound 6J was prepared according to the following scheme:

Step 1: Preparation of Oi-/('/7- butyl Oa-methyl 4-but-3-enylpiperidine-l,4- dicarboxylate (compound 6C)

To a solution of Oi-tert-butyl CU-methyl piperidine- 1,4-dicarboxylate (compound 6A, 5.0 g, 20.55 mmol) in THF (80 mL) was added LDA (11.3 mL, 22.61 mmol) at -70 °C under nitrogen atmosphere. After being stirred for 0.5 h, the reaction mixture was added with 4-bromobut-l-ene (compound 6B, 3.0 g, 22.61 mmol) and the resulting mixture was warmed up to 20 °C and stirred for another 2.5 hrs. After the reaction was completed, it was quenched by addition of sat. NH4CI aq. at 0 °C and the reaction mixture was extracted with EtOAc (50 mL, twice). The combined organic layer was washed with brine, dried over Na2SO4, filtered and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE = 5 % to 30 %) to afford Oi-tert-butyl O4- methyl 4-but-3-enylpiperidine- 1,4-dicarboxylate (compound 6C, 4.13 g) as yellow oil^H NMR (400 MHz, CHLOROFORM-<7) 8 5.69 - 5.81 (m, 1 H), 4.91 - 5.06 (m, 2 H), 3.87 (d, J = 14.0 Hz, 2 H), 3.71 (s, 3 H), 2.92 - 2.82 (m, 2 H), 2.11 (d, J = 13.6 Hz, 2 H), 2.00 - 1.92 (m, 2 H), 1.63 - 1.58 (m, 2 H), 1.45 (s, 9 H), 1.40 - 1.32 (m, 2 H) ppm.

Step 2: Preparation of Oi-/('/7-butyl 04-methyl 4-(3-oxopropyl)piperidine-l,4- dicarboxylate (compound 6D) To a mixture of Oi-/c/7-butyl O4- methyl 4-but-3-enylpiperidine-l,4-dicarboxylate (compound 6C, 1.9 g, 6.49 mmol) in 1,4-dioxane (40 mL) and water (4 mL) was added 2,6- Lutidine (1.5 mL, 12.98 mmol) and potassium osmate (VI) dihydrate (119.5 mg, 0.32 mmol) at 0 °C. After being stirred at 0 °C for 15 min, the reaction mixture was added with sodium metaperiodate (5.6 g, 25.96 mmol) at 0 °C and the resulting mixture was warmed to 20 °C and stirred for 3 hrs. After the reaction was completed, the reaction mixture was extracted with EtOAc (20 mL, three times), and washed with saturated Na2SOs aqueous solution (20 mL). The combined organic layer was washed with brine (20 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to afford Oi-/e/7-butyl CL- methyl 4-(3-oxopropyl)piperidine-l,4- dicarboxylate (compound 6D, 2.0 g) as yellow oil. MS calc’d 200.2 (M-Boc+H ⁺ ), measured 200.1 (M-Boc+H ⁺ ).

Step 3: Preparation of Oi-/c/7- butyl (h-methyl 4-[3-[[( IS)- l-tert-butoxycarbonyl-2- methyl-propyl]amino]propyl]piperidine-l,4-dicarboxylate (compound 6F)

To a mixture of Oi-/e/7-butyl O4- methyl 4-(3-oxopropyl)piperidine-l,4-dicarboxylate (compound 6D,1.0 g, 3.34 mmol) and Boc-V-Me-Val-OH (770.5 mg, 3.67 mmol) in methanol (20 mL) was added zinc chloride (500.7 mg, 3.67 mmol) at 0 °C. After being was stirred at 0 °C for 0.5 h, the reaction mixture was added with sodium cyanoborohydride (272.8 mg, 4.34 mmol) at 0 °C and the resulting mixture was warmed to 20 °C and stirred for another 1 h. After the reaction was completed, it was quenched with sat. NH4CI aq. (20 mL) and the reaction mixture was extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (20 mL), dried with anhydrous Na2SO4 , filtered and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE = 10 % to 30 %) to afford 01 -/c/7- butyl O4-methyl 4-[3-[ [(IS)- l-/c/7-butoxycarbony 1-2- methylpropyl] amino] propyl] piperidine- 1,4-dicarboxy late (compound 6F, 1.1 g) as yellow oil. MS calc’d 457.3 (MH ⁺ ), measured 457.2 (MH ⁺ ).

Step 4: Preparation of lithium; 1 -/c/7-butoxycarbonyl-4-|3-||( IS)- butoxycarbonyl-2-methyl-propyl]amino]propyl]piperidine-4-car boxylate (compound 6G)

To a mixture of Oi-tert-butyl O4-methyl 4-[3-[[(lS)-l-tert-butoxycarbonyl-2-methyl- propyl]amino]propyl]piperidine- 1,4-dicarboxy late (compound 6F, 400.0 mg, 0.88 mmol) in methanol (6 mL), THF (0.6 mL) and water (0.6 mL) was added lithium hydroxide monohydrate (367.6 mg, 8.76 mmol). The resulting mixture was heated to 60 °C and stirred for 12 hrs. After the reaction was completed, the reaction mixture was cooled to 20 °C and concentrated under vacuum to afford lithium;l-tert-butoxycarbonyl-4-[3-[[(lS)-l-tert-butoxycarbo nyl-2-methyl- propyl]amino]propyl]piperidine-4-carboxylate (compound 6G, 392.0 mg) as a white solid. MS calc’d 443.3 (M-Li+H ⁺ ), measured 443.2 (M-Li+H ⁺ ).

Step 5: Preparation of tert-butyl 2-[(lS)-l-tert-butoxycarbonyl-2-methyl-propyl]-l- oxo-2, 9-diazaspiro[5.5]undecane-9-carboxylate (compound 6H)

To the mixture of lithium;l-tert-butoxycarbonyl-4-[3-[[(lS)-l-tert-butoxycarbo nyl-2- methyl-propyl]amino]propyl]piperidine-4-carboxylate (392.0 mg, 0.87 mmol) in DMF (8 mL) was added DIEA (0.5 mL, 2.62 mmol) and COMU (673.7 mg, 1.57 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was poured into water (80 mL) and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (20 mL, three times), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE = 5% to 25%) to afford tert-butyl 2-[(15)-l-tert-butoxycarbonyl-2-methyl-propyl]- l-oxo-2,9-diazaspiro[5.5]undecane-9-carboxylate (compound 6H, 200.0 mg) as yellow oil. MS calc’d 425.3 (MH ⁺ ), measured 425.3 (MH ⁺ ).

Step 6: Preparation of (2S)-3-methyl-2-(l-oxo-2,9-diazaspiro[5.5]undecan-2- yl)butanoic acid (compound 61)

To a solution of tert-butyl 2-[(15)-l-tert-butoxycarbonyl-2-methyl-propyl]-l-oxo-2,9- diazaspiro[5.5]undecane-9-carboxylate (compound 6H, 200.0 mg, 0.47 mmol) in DCM (1.5 mL) was added trifluoro acetic acid (1.0 mL, 12.98 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the mixture was concentrated under vacuum to give (25)-3- methyl-2-(l-oxo-2,9-diazaspiro[5.5]undecan-2-yl)butanoic acid (compound 61, 180.0 mg, TFA salt) as colorless oil. MS calc’d 269.2 (MH ⁺ ), measured 269.2 (MH ⁺ ).

Step 7: Preparation of (2.S )-2-(9-/er/-butoxycarbonyl-l-oxo-2,9- diazaspiro[5.5]undecan-2-yl)-3-methyl-butanoic acid (compound 6J)

To a solution of (2S)-3-methyl-2-(l-oxo-2,9-diazaspiro[5.5]undecan-2-yl)butan oic acid (compound 61, 180.0 mg, 0.47 mmol, TFA salt) in THF (3 mL) and water (3 mL) was added sodium carbonate (99.8 mg, 0.94 mmol) and di-tert-butyl dicarbonate (123.3 mg, 0.56 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the mixture was concentrated under vacuum to give a residue, which was purified by reversed phase flash column to afford (2S)-2-(9-tert-butoxycarbonyl- l-oxo-2,9-diazaspiro[5.5]undecan-2-yl)-3-methyl- butanoic acid (compound 6J, 400.0 mg) as an off-white solid. MS calc’d 367.2 (MH ), measured 367.2 (MH’).

Example 8 (3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(8S,14S) -25-fluoro-4-hydroxy-(21M)-21- [2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridy l]-18,18-dimethyl-9,15-dioxo- 22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacydo[18 .5.2.1 ² ’ ⁶ .l ¹⁰ ’ ¹⁴ .0 ²³ ’ ²⁷ ]nonacosa- l(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methy l-propyl]-2V-methyl- pyrrolidine-3-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (8S,14S)-8-amino-25-fluoro-4-hydroxy-(21A7)-21-[2-[(15')-l-m ethoxyethyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl]- 18, 18-dimethyl-22-(2, 2, 2-trifluoroethyl)- 16-oxa- 10,22,28- triazapentacyc lo[ 18.5.2.1 ² ’ ⁶ . l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa- 1(26), 2, 4, 6(29), 20, 23(27), 24-heptaene-9, 15- dione (Intermediate G) instead of (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate E). Example 8 (21.6 mg) was obtained as a white solid. MS calc’d 1114.5 (MEE), measured 1114.8 (MIE). ^J H NMR (400 MHz, Methanol-^) 6 = 8.43 (d, J = 2.4 Hz, 1H), 7.62 (dd, J = 10.8, 7.2 Hz, 1H), 7.55 - 7.53 (m, 2H), 7.29 (s, 1H), 7.26 - 7.20 (m, 1H), 6.96 - 6.90 (m, 1H), 6.65 - 6.55 (m, 1H), 5.65 - 5.49 (m, 1H), 4.97 - 4.68 (m, 2H), 4.67 - 4.62 (m, 1H), 4.46 - 4.38 (m, 1H), 4.20 - 4.12 (m, 1H), 4.00 - 3.87 (m, 1H), 3.82 - 3.61 (m, 6H), 3.59 - 3.43 (m, 5H), 3.38 - 3.32 (m, 2H), 3.25 - 3.11 (m, 1H), 3.04 (s, 2H), 2.99 - 2.96 (m, 1H), 2.93 (s, 4H), 2.90 - 2.65 (m, 3H), 2.63 - 2.47 (m, 2H), 2.39 - 2.11 (m, 3H), 2.10 - 1.95 (m, 2H), 1.89 - 1.79 (m, 1H), 1.58 - 1.27 (m, 6H), 1.03 - 0.92 (m, 6H), 0.91 - 0.84 (m, 1H), 0.83 - 0.75 (m, 3H), 0.47 - 0.32 (m, 3H) ppm. Example 9 l-[(2/?)-2-chloro-2-fluoro-acetyl]-4-fluoro-^-[(lS)-l-[[(7S, 13S) -(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17,24 -trimethyl-8,14-dioxo-21- (2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-A^-methyl-piperidine-4-carbox amide

The title compound was prepared in analogy to the preparation of Example 1 by using 1- tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-(20Af)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17,24-trimethyl-21-(2,2,2- 25 9 13 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate F) instead of (S)-l-Boc- pyrrolidine-3-carboxylic acid (compound ID) and (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E). Example 9 (2.7 mg) was obtained as a white solid. MS calc’d 1133.5 (MEE), measured 1133.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.49 - 8.36 (m, 1H), 8.03 (d, J = 13.6 Hz, 1H), 7.58 - 7.39 (m, 3H), 7.12 - 6.94 (m, 1H), 5.90 - 5.77 (m, 1H), 5.26 - 5.11 (m, 1H), 4.73 - 4.61 (m, 1H), 4.52 - 4.31 (m, 2H), 4.21 (m, 1H), 4.14 - 3.84 (m, 4H), 3.75 - 3.49 (m, 3H), 3.26 - 3.07 (m, 10H), 3.00 (s, 4H), 2.52 (m, 6H), 2.15 (m, 8H), 1.97 - 1.82 (m, 1H), 1.71 - 1.50 (m, 2H), 1.45 (m, 3H), 1.36 - 1.24 (m, 3H), 1.02 - 0.87 (m, 9H), 0.39 (s, 3H) ppm.

Example 10 l-[(2/?)-2-chloro-2-fluoro-acetyl]-4-fluoro-^-[(lS)-l-[[(7S, 13S)-24-fluoro-(20M)-20-[2-[(lS)- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-A^-methyl-piperid ine-

4-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using 1- tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-24-fluoro- (20A7)-20-[2-[(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl )-3-pyridyl]-17,17-dimethyl-21- 25 9 13 22226 (2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapenta cyclo[17.5.2.1 ’ .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D) instead of (S)-l-Boc- pyrrolidine-3-carboxylic acid (compound ID) and (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E). Example 10 (17.5 mg) was obtained as a yellow solid. MS calc’d 1137.5 (MEE), measured 1137.5 (MH ⁺ ), ^J H NMR (400 MHz, Methanol-^) 6 = 8.74 - 8.68 (m, 1H), 8.54 - 8.51 (m, 1H), 7.74 - 7.70 (m, 1H), 7.57 - 7.47 (m, 2H), 7.15 - 6.97 (m, 1H), 5.75 - 5.64 (m, 1H), 5.26 - 5.12 (m, 1H), 4.83 - 4.73 (m, 1H), 4.51 - 4.32 (m, 2H), 4.29 - 4.21 (m, 2H), 4.20 - 3.89 (m, 3H), 3.88 - 3.62 (m, 5H), 3.60 - 3.43 (m, 4H), 3.40 - 3.36 (m, 4H), 3.26 - 3.22 (m, 4H), 3.04 - 3.00 (m, 4H), 2.89 - 2.81 (m, 1H), 2.64 - 2.56 (m, 1H), 2.39 - 2.15 (m, 6H), 1.98 - 1.95 (m, 1H), 1.89 - 1.78 (m, 1H), 1.73 - 1.59 (m, 1H), 1.50 - 1.43 (m, 4H), 1.38 - 1.28 (m, 1H), 1.04 - 0.97 (m, 6H), 0.95 - 0.88 (m, 3H), 0.51 - 0.41 (m, 3H) ppm.

Example 11 (2S)-2-[2-[(2/f)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazasp iro[3.5]nonan-6-yl]-2V-[(7S,13S)- 24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpipe razin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.5]nonan -6-yl)-3-methyl-butanoic acid (compound 11J) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J). Example 11 (6.6 mg) was obtained as a white solid. MS calc’d 1117.4 (MH ⁺ ), measured 1117.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.68 (d, J = 7.3 Hz, 1H), 8.50 (d, J = 2.8 Hz, 1H), 7.72 (dd, J = 2.4, 15.6 Hz, 1H), 7.50 - 7.45 (m, 2H), 6.79 - 6.60 (m, 1H), 5.72 - 5.64 (m, 1H), 5.21 - 5.10 (m, 3H), 4.44 (br s, 2H), 4.21 (br d, J = 5.6 Hz, 3H), 4.05 - 3.86 (m, 3H), 3.79 (br d, J = 11.2 Hz, 1H), 3.74 - 3.61 (m, 4H), 3.57 - 3.40 (m, 8H), 3.35 (br s, 2H), 3.15 (br d, J = 14.8 Hz, 3H), 3.00 (s, 3H), 2.83 (br t, J = 11.6 Hz, 1H), 2.62

- 2.50 (m, 1H), 2.20 (br s, 3H), 1.99 - 1.94 (m, 1H), 1.83 (br s, 3H), 1.64 (br dd, J = 4.0, 12.0 Hz, 1H), 1.44 (d, J = 6.0 Hz, 3H), 1.00 (br dd, J = 2.8, 6.4 Hz, 3H), 0.97 (s, 3H), 0.89 (br d, J = 6.4 Hz, 3H), 0.44 (s, 3H) ppm.

The compound 11J was prepared according to the following scheme:

111 11J

Step 1: Preparation of tert-butyl 3-but-3-enyl-3-cyano-azetidine-l-carboxylate (compound 11B)

To a solution of tert-butyl 3-cyanoazetidine-l-carboxylate (compound 11A, 5.0 g, 27.44 mmol) in THF (50 mL) cooled to -70 °C was added lithium bis(trimethylsilyl) amide (30.1 mL, 30.18 mmol) dropwise under nitrogen atmosphere. After being stirred for 0.5 h, the reaction mixture was added with 4-bro mo- 1 -butene (4.4 g, 32.93 mmol) slowly at -70 °C. The mixture was warmed up to 20 °C and stirred for 3 hrs. After the reaction was completed, it was quenched with saturated NH4CI (200 mL) and the reaction mixture was extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE = 5 % to 30 %) to afford tert-butyl 3-but-3-enyl-3-cyano- azetidine-1 -carboxylate (compound 11B, 5.4 g) as yellow oil. MS calc’d 237.2 (MH ⁺ ), measured 181.6 (M-C ₄ H ₈ +H ⁺ ).

Step 2: Preparation of 3-but-3-enyl- l-/c/7-butoxycarbonyl-azetidine-3-carboxylic add (compound 11C)

To the mixture of tert-butyl 3-but-3-enyl-3-cyano-azetidine-l-carboxylate (compound 11B, 5.4 g, 22.85 mmol) in ethanol (60 mL) and water (60 mL) was added potassium hydroxide (5.1 g, 91.41 mmol) in one portion. The mixture was heated at 80 °C for 16 hrs. After the reaction was completed, the pH of the reaction mixture was adjust to pH=3 with IN HC1 aqueous solution and it was extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (100 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to give 3- but-3-enyl-l-tert-butoxycarbonyl-azetidine-3-carboxylic acid (compound 11C, 6.31 g) as yellow oil. MS calc’d 256.2 (MH ⁺ ), measured 200.3 (M-C ₄ H ₈ +H ⁺ ).

Step 3: Preparation of Oi-/c/7- butyl (L-methyl 3-but-3-enylazetidine-l,3- dicarboxylate (compound 11D)

To the mixture of 3-but-3-enyl-l-tert-butoxycarbonyl-azetidine-3-carboxylic acid (compound 11C, 6.3 g, 24.72 mmol) in DMF (60 mL) was added potassium carbonate (10.2 g, 74.15 mmol) and iodomethane (7.0 g, 49.43 mmol) in one portion. The mixture was stirred at 20 °C for 2 hrs. After the reaction was completed, the mixture was poured into water (150 mL) and extracted with EtOAc (70 mL, three times). The combined organic layer was washed with brine (70 mL, three times), dried with anhydrous Na2SO4, filtered and concentrated under vacuum. The residue was purified by column chromatography (EtOAc in PE = 5 % to 30 %) to afford Oi- tert-butyl Os-methyl 3-but-3-enylazetidine-l,3-dicarboxylate (compound 11D, 3.76 g) as yellow oil. MS calc’d 270.2 (MH ⁺ ), measured 214.3 (M-C ₄ H ₈ +H ⁺ ).

Step 4: Preparation of Oi-tert-butyl O3-methyl 3-(3-oxopropyl)azetidine-l,3- dicarboxylate (compound HE)

To the mixture of Oi-tert-butyl Ovincthyl 3-but-3-enylazetidine-l,3-dicarboxylate (compound HD, 3.7 g, 13.96 mmol) in 1,4-dioxane (100 mL) and water (10 mL) was added 2,6- Lutidine (3.2 mL, 27.92 mmol) and potassium osmate(VI) dihydrate (0.3 g, 0.7 mmol) in one portion at 0 °C. After being stirred at 0 °C for 15 minutes, the reaction mixture was added with sodium metaperiodate (11.9 g, 55.84 mmol) and the resulting mixture was warmed to 20 °C and stirred for another 3 hrs. After the reaction was completed, the reaction was quenched by addition with saturated Na2SOs aqueous solution (200 mL) and the reaction mixture was extracted with EtOAc (100 mL, three times). The combined organic layer was washed with brine (100 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to afford Oi- tert-butyl CL-methyl 3-(3-oxopropyl)azetidine-l,3-dicarboxylate (compound HE, 3.8 g) as yellow oil. MS calc’d 272.2 (MH ⁺ ), measured 216.3 (M-C4H ₈ +H ⁺ ).

Step 5: Preparation of Oi-/c/7- butyl Ch-methyl 3-[3-[[(lS)-l-tert-butoxycarbonyl-2- methyl-propyl]amino]propyl]azetidine-l,3-dicarboxylate (compound HF)

To the mixture of tert-butyl- L-valinate hydrogen chloride (1.8 g, 10.5 mmol) and Oi-tert- butyl Ch-methyl 3-(3-oxopropyl)azetidine-l,3-dicarboxylate (compound HE, 1.9 g, 7.0 mmol) in Methanol (20 mL) was added zinc chloride (1.4 g, 10.5 mmol) in one portion at 0 °C. After being stirred at 20 °C for 0.5 h, the reaction mixture was added with sodium cyanoborohydride (0.9 g, 14.01 mmol) at 0 °C and the resulting mixture was stirred at 20 °C for another 1 h. After the reaction was completed, it was quenched by addition of saturated NH4CI aqueous solution (100 mL) and the reaction mixture was extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (50 mL), dried with anhydrous Na2SO4 , filtered and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE = 10 % to 30 %) to afford Oi-tert-butyl Os-methyl 3-[3-[[( IS)- 1- tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]azetidine-l ,3-dicarboxylate (compound HE, 1.2 g) as yellow oil. MS calc’d 429.3 (MH ⁺ ), measured 429.4 (MH ⁺ ).

Step 6: Preparation of lithium; 1 -/c/7-butoxycarbonyl-3-|3-||( l.S)-l-/(T/- butoxycarbonyl-2-methyl-propyl]amino]propyl]azetidine-3-carb oxylate (compound HG)

To the mixture of Oi-tert-butyl Os-methyl 3-[3-[[(lS)-l-tert-butoxycarbonyl-2-methyl- propyl]amino]propyl]azetidine-l,3-dicarboxylate (compound HE, 1.2 g, 2.8 mmol) in methanol (30 mL), water (3 mL) and THF (3 mL) was added lithium hydroxide monohydrate (1.2 g, 28.0 mmol). The mixture was then heated to 60 °C and stirred for 12 hrs. After the reaction was completed, the reaction mixture was concentrated under vacuum to afford lithium;l -/<?/7- butoxycarbonyl-3-[3-[[(lS)-l-tert-butoxycarbonyl-2-methyl-pr opyl]amino]propyl]azetidine-3- carboxylate (compound 11G, 2.3 g) as a yellow solid. MS calc’d 421.8 (MH ⁺ ), measured 415.2 (M-Li+H ⁺ ).

Step 7: Preparation of tert-butyl 6-[(lS)-l-tert-butoxycarbonyl-2-methyl-propyl]-5- oxo-2, 6-diazaspiro[3.5]nonane-2-carboxylate (compound 11H)

To a solution of lithium;l-tert-butoxycarbonyl-3-[3-[[(lS)-l-tert-butoxycarbo nyl-2- methyl-propyl]amino]propyl]azetidine-3 -carboxylate (compound 11G, 2.0 g, 2.41 mmol) in DMF (30 mL) was added DIEA (2.1 mL, 12.06 mmol) and COMU (2.0 g, 4.82 mmol) at 0 °C in one portion. The reaction mixture was then warmed up to 20 °C and stirred for 1 h. After the reaction was completed, the mixture was poured into water (100 mL) and extracted with EtOAc (50mL, three times). The combined organic layer was washed with brine (50 mL, three times), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE = 20 % to 30 %) to afford tert-butyl 6- [(lS)-l-tert-butoxycarbonyl-2-methyl-propyl]-5-oxo-2,6-diaza spiro[3.5]nonane-2-carboxylate (compound 11H, 512.0 mg) as a yellow solid. MS calc’d 397.3 (MH ⁺ ), measured 419.4 (MNa ⁺ ).

Step 8: Preparation of (2S)-3-methyl-2-(5-oxo-2,6-diazaspiro[3.5]nonan-6-yl)butanoi c add (compound 111)

To a solution of tert-butyl 6-[(15)-l-tert-butoxycarbonyl-2-methyl-propyl]-5-oxo-2,6- diazaspiro[3.5]nonane-2-carboxylate (compound 11H, 512.0 mg, 1.29 mmol) in DCM (3 mL) was added TFA (3.0 mL, 38.94 mmol). The mixture was stirred at 20 °C for 2 hrs. After the reaction was completed, the mixture was concentrated under vacuum to afford (2S)-3-methyl-2- (5-oxo-2,6-diazaspiro[3.5]nonan-6-yl)butanoic acid (compound 111, 301.0 mg) as yellow oil as TFA salt. MS calc’d 241.2 (MH ⁺ ), measured 241.5 (MH ⁺ ).

Step 9: Preparation of (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.5]nonan - 6-yl)-3-methyl-butanoic acid (compound 11J)

To the mixture of (2S)-3-methyl-2-(5-oxo-2,6-diazaspiro[3.5]nonan-6-yl)butanoi c acid (compound 111, 301.0 mg, 1.25 mmol) in THF (12 mL) was added di-t-butyldicarbonate (326.9 mg, 1.5 mmol) and sodium carbonate (661.6 mg, 6.24 mmol) in one portion, the mixture was stirred at 20 °C for 2 hrs. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified by prep-HPLC to afford (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.5]nonan -6-yl)-3-methyl-butanoic acid (compound 11J, 303.0 mg) as colorless gum. MS calc’d 341.2 (MH ⁺ ), measured 363.2 (MNa ⁺ ).

Example 13

(2/?)-2-[[l-[(2/?)-2-chloro-2-fluoro-acetyl]azetidin-3-yl ]oxymethyl]-A^-[(7S,13S)-24-fluoro- (20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl )-3-pyridyl]-17,17-dimethyl- 8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,2 8-tetrazapentacyclo- [17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-me thyl-butanamide

The compound was prepared according to the following scheme:

Step 1: Preparation of ethyl 3-hydroxy-3-methyl-2-methylene-butanoate (compound

13C)

DIBAL-H (152.9 mL, 152.91 mmol) was added dropwise at 0 °C to a solution of NMO (23.8 g, 203.87 mmol) in THF (500 mL). After being stirred at 0 °C for 30 min, the reaction mixture was added with ethyl prop-2-ynoate (compound 13A, 10 g, 101.94 mmol) dropwise at 0 °C. After being stirred at 0 °C for another 1 h, the reaction mixture were added with acetone (compound 13B, 5.9 g, 101.94 mmol) and boron trifluoride etherate (12.9 mL, 101.94 mmol) and the resulting mixture was stirred at room temperature for 18 hrs. The reaction was worked up by addition of EA (100 mL). The organic layer was washed with HC1 (IN, 37.5 mL), sat. NaHCCh (10 mL) and water (10 mL), and then dried over Na2SO4 and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (PE: EA=10:l) to afford ethyl 3- hydroxy-3-methyl-2-methylene-butanoate (compound 13C, 4.8 g) as colorless oil. ^J H NMR (400 MHz, CHLOROFORM-<7) 5 = 6.14 (s, 1H), 5.83 - 5.70 (m, 1H), 4.23 (q, 7 = 7.3 Hz, 2H), 3.93 (br s, 1H), 1.49 - 1.37 (m, 6H), 1.31 (t, 7 = 7.1 Hz, 3H) ppm.

Step 2: Preparation of ethyl 2-(bromomethyl)-3-methyl-but-2-enoate (compound 13D)

To a solution of ethyl 3-hydroxy-3-methyl-2-methylene-butanoate (compound 13C, 4.8 g, 30.34 mmol) in THF (100 mL) was added dropwise a solution of Phosphorus tribromide (4.9 g, 18.21 mmol) at 0 °C. After being stirred at 15 °C for 2.5 hrs, the reaction mixture was poured into ice water at 0 °C, and the aqueous layer was extracted with EA (10 mL). The organic layer was washed with brine, dried over Na2SO4, and concentrated to give the crude product, which was purified by silica gel chromatography (petroleum/EtOAc=20/l~10/l) to afford ethyl 2- (bromomethyl)-3-methyl-but-2-enoate (compound 13D, 5.3 g) as colorless oil. ¹ H NMR (400 MHz, CHLOROFORM-<7) 8 = 4.30 (s, 2H), 4.24 (d, J = 7.2 Hz, 2H), 2.15 (s, 3H), 1.97 (s, 3H), 1.32 (t, 7 = 7.2 Hz, 3H) ppm.

Step 3: Preparation of benzyl 3-(2-ethoxycarbonyl-3-methyl-but-2-enoxy)azetidine-l- carboxylate (compound 13F)

To a mixture of benzyl 3 -hydroxyazetidine- 1 -carboxylate (compound 13E, 625.0 mg, 3.02 mmol), tetrabutylammonium iodide (116.0 mg, 0.31 mmol) and sodium hydroxide (267.8 mg, 6.7 mmol) in Water (2 mL) and THF (4 mL) was added ethyl 2-(bromomethyl)-3-methyl-but-2- enoate (compound 13D, 1.0 g, 4.52 mmol) at 0 °C. After being stirred at 10 °C for 16 hrs, the reaction mixture was dilute with water (5 mL) and the resulting mixture was extracted with EA (10 mL, twice). The combined organic layer was washed with brine (10 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum/EtOAc=20/l~10/l) to afford benzyl 3-(2-ethoxycarbonyl-3-methyl-but-2- enoxy)azetidine-l -carboxylate (compound 13F, 590.0 mg) as colorless oil. MS calc’d 348.1 (MH ⁺ ), measured 348.1 (MH ⁺ ).

Step 4: Preparation of tert-butyl 3-(2-ethoxycarbonyl-3-methyl-butoxy)azetidine-l- carboxylate (compound 13G)

To a solution benzyl 3-(2-ethoxycarbonyl-3-methyl-but-2-enoxy)azetidine-l-carboxy late (compound 13F, 950.0 mg, 2.73 mmol) in Methanol (25 mL) was added Pd/C (240.0 mg, 0.26 mmol) at 15 °C under N2. The reaction was purged with H2 and stirred at 50 °C for 2 hrs under H2 atmosphere. The reaction mixture was filtered and the filter cake was washed with methanol (30 mL). The combined filtrate was concentrated under vacuum and the resulting residue was purified by silica gel chromatography (petroleum/EtOAc=10/l~5/l) to afford tert-butyl 3-(2- ethoxycarbonyl-3-methyl-butoxy)azetidine-l-carboxylate (compound 13G, 700.0 mg) as colorless oil. ^J H NMR (400 MHz, CHLOROFORM-<7) 8 = 4.25 - 4.11 (m, 3H), 4.08 - 3.99 (m, 2H), 3.80 (dd, J = 4.3, 9.4 Hz, 2H), 3.61 - 3.46 (m, 2H), 2.45 (ddd, J = 4.9, 7.3, 9.5 Hz, 1H), 1.97 - 1.86 (m, 1H), 1.43 (s, 9H), 1.31 - 1.24 (m, 3H), 0.99 - 0.90 (m, 6H) ppm.

Step 5: Preparation of 2-|( l-/er/-butoxycarbonylazetidin-3-yl)oxyniethyl]-3-methyl- butanoic acid (compound 13H)

To a mixture of tert-butyl 3-(2-ethoxycarbonyl-3-methyl-butoxy)azetidine-l-carboxylate (compound 13G, 350.0 mg, 1.11 mmol) in methanol (6 mL) was added NaOH aq. (I N, 6.0 mL, 6.0 mmol). After being stirred at 50 °C for 16 hrs, the reaction mixture was concentrated under vacuum. The resulting residue was diluted with water (10 mL), and the pH of the reaction mixture was adjusted pH=5 by IN HC1 aq., then the reaction mixture was extracted with EA (30 mL, twice). The combined organic layer was washed with brine (50 mL), dried (Na2SO4) and concentrated under vacuum to give crude product. The crude was purified by silica gel chromatography (petroleum/EtOAc=5/l~3/l) to afford 2-[( l-/c/7-butoxycarbonylazctidin-3- yl)oxymethyl]-3-methyl-butanoic acid (compound 13H, 200.0 mg) as colorless oil. ¹ H NMR (400 MHz, CHLOROFORM-d) 8 = 4.22 (tt, J = 4.3, 6.4 Hz, 1H), 4.10 - 4.03 (m, 2H), 3.83 (dd, J = 4.3, 9.7 Hz, 2H), 3.63 - 3.48 (m, 2H), 2.50 (ddd, J = 4.8, 7.1, 9.2 Hz, 1H), 1.99 (qd, J = 6.8, 13.7 Hz, 1H), 1.44 (s, 9H), 0.99 (dd, J = 2.9, 6.8 Hz, 6H) ppm.

Step 6: Preparation of tert-butyl 3-[(2/?)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2- 25 9 13 2226 trifhioroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ Joctacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carbox ylate (compound 131) & tert-butyl 3-[(2S)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carbox ylate

(compound 13J)

To a solution of (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')-l-methoxyethy l]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate D, 225.0 mg, 0.28 mmol) and 2-[(l-tert-butoxycarbonylazetidin-3- yl)oxymethyl]-3-methyl-butanoic acid (compound 13H, 104.9 mg, 0.37 mmol) in DMF (2.5 mL) were added DIEA (0.2 mL, 0.84 mmol) and HATU (213.6 mg, 0.56 mmol) at 0 °C. After being stirred at 20 °C for 1 h, the reaction mixture was diluted with water (3 mL), extracted with EtOAc (8 mL, twice). The combined organic layer was washed with brine (10 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by prep-HPLC to afford tert-butyl 3-[(2/?)-2-[[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l-methoxy ethyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carboxyla te (compound 131, faster eluted, 45.0 mg) as a light yellow solid and tert-butyl 3-[(25 ^, )-2-[[(7S,13S)-24-fluoro-(20A7)-20-[2-[(15')- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-

25 9 13 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carbox ylate (compound 13J, slower eluted, 100.0 mg) as a light yellow solid. MS calc’d 1070.6 (MH ⁺ ), measured 1070.6 (MH ⁺ ).

Step 7: Preparation of (2/f)-2-(azetidin-3-yloxymethyl)-A^-[(7S,13S)-24-fluoro-(20A f)- 20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyr idyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (compound 13K)

To a solution of tert-butyl 3-[(2/?)-2-[[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carbox ylate (compound 131, 45.0 mg, 0.04 mmol) in DCM (0.5 mL) was added TFA (0.2 mL) at 0 °C, then the mixture was stirred at 15 °C for 0.5 h. The reaction mixture was concentrated under vacuum to afford (2R)-2-(azetidin-3-yloxymethyl)-A-[(7S,13S)-24-fluoro-(20A7) -20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2-

25 9 13 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-butanamide; 2,2,2-trifluoroacetic acid (compound 13K, 40.0 mg) as yellow oil. MS calc’d 970.4 (MH ⁺ ), measured 970.4 (MH ⁺ ).

Step 8: Preparation of (2/f)-2-[[l-[(2/f)-2-chloro-2-fluoro-acetyl]azetidin-3- yl]oxymethyl]-^-[(7S,13S)-24-fluoro-(20M)-20-[2-[(lS)-l-meth oxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-2 1-(2,2,2-trifluoroethyl)-15- oxa-4-thia-9,21,27,28-tetrazapentacyclo-[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-butanamide (Example 13)

To a solution of (2R)-2-(azetidin-3-yloxymethyl)-A-[(7S,13S)-24-fluoro-(20Af) -20-[2- [(15)- l-methoxyethyl]-5-(4-methylpiperazin- 1 -yl) - 3 -pyridyl] - 17, 17-dimethyl-8, 14-dioxo-21- 25 9 13 2226 (2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapenta cyclo[17.5.2.1 ’ .1 .0 ’ ]octacosa- l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-butanamide; 2,2,2-trifluoroacetic acid (compound 13K, 40.0 mg, 0.04 mmol) in DMF (0.2 mL) were added DIEA (0.1 mL, 0.74 mmol), T3P (469.6 mg, 0.74 mmol) and (2R)-2-chloro-2-fluoro-acetic acid (compound 13L, 83.0 mg, 0.74 mmol) at 0 °C. After being stirred at 20 °C for 1 h, the reaction mixture was concentrated under vacuum and the resulting residue was purified by prep-HPLC to afford (2R)-2-[[l-[(2R)-2- chloro-2-fluoro-acetyl]azetidin-3-yl]oxymethyl]-A-[(7S,13S)- 24-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo-[ 17.5.2.1 ’ .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-butanamide (Example 13, 22.6 mg) as a white solid. MS calc’d 1064.5 (MH ⁺ ), measured 1064.5 (MH*). ¹ !! NMR (400 MHz, CHLOROFORM- <7) 6 = 8.65 (br d, 7 = 6.8 Hz, 2H), 7.64 (dd, J = 1.9, 4.9 Hz, 1H), 7.26 - 7.20 (m, 1H), 7.14 (br d, J= 11.9 Hz, 1H), 6.77 - 6.67 (m, 1H), 6.41 - 6.22 (m, 1H), 5.89 (br t, J = 8.1 Hz, 1H), 4.95 (br dd, J = 9.0, 16.5 Hz, 1H), 4.65 - 4.48 (m, 3H), 4.46 - 4.21 (m, 6H), 4.17 - 4.09 (m, 1H), 3.86 (br d, J = 11.1 Hz, 2H), 3.74 - 3.69 (m, 3H), 3.60 - 3.55 (m, 2H), 3.52 - 3.45 (m, 2H), 3.36 (s, 3H), 3.15 - 3.09 (m, 2H), 2.94 - 2.87 (m, 3H), 2.75 - 2.67 (m, 1H), 2.47 - 2.34 (m, 1H), 2.30 - 2.21 (m, 2H), 2.04 - 1.94 (m, 4H), 1.88 - 1.79 (m, 1H), 1.70 - 1.54 (m, 2H), 1.50 (d, 7 = 6.1 Hz, 3H), 1.30 - 1.20 (m, 1H), 1.02 (dd, 7 = 4.6, 6.3 Hz, 6H), 0.97 (s, 3H), 0.45 (s, 3H) ppm. Example 14

(2S)-2-[[l-[(2/f)-2-chloro-2-fluoro-acetyl]azetidin-3-yl] oxymethyl]-AH(7S,13S)-24- fluoro- (20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl )-3-pyridyl]-17,17-dimethyl- 8,14-dioxo-21-(2,2,2-trifhioroethyl)-15-oxa-4-thia-9,21,27,2 8- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 13 by using fert-butyl 3-[(25)-2-[[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l-methoxye thyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-2 1-(2,2,2-trifluoroethyl)-15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carboxyla te (compound 13J) instead of fert-butyl 3-[(2/?)-2-[[(75,135)-24-fluoro-(20Af)-20-[2-[(15)-l-methoxy ethyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl-8, 14-dioxo-21-(2,2,2-trifluoroethyl)- 15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carboxyla te (compound 131). Example 14 (38.0 mg) was obtained as a white solid. MS calc’d 1064.5 (MEE), measured 1064.5 (MH ⁺ ). ^J H NMR (400 MHz, CHLOROFORM-<7) 8 = 8.72 (br s, 1H), 8.66 (d, J = 7.4 Hz, 1H), 7.65 (s, 1H), 7.14 (d, J = 12.0 Hz, 1H), 6.80 - 6.72 (m, 1H), 6.39 - 6.23 (m, 1H), 5.97 - 5.86 (m, 1H), 5.01 - 4.83 (m, 2H), 4.60 - 4.49 (m, 5H), 4.36 - 4.22 (m, 6H), 4.05 - 3.93 (m, 2H), 3.87 (br d, J = 10.4 Hz, 1H), 3.73 - 3.68 (m, 2H), 3.64 - 3.53 (m, 4H), 3.45 (s, 1H), 3.36 (s, 3H), 3.20 - 3.00 (m, 4H), 2.93 (br s, 3H), 2.73 (br t, J = 13.2 Hz, 1H), 2.43 (br d, J = 14.1 Hz, 1H), 2.30 - 2.20 (m, 2H), 2.04 - 1.96 (m, 2H), 1.85 - 1.78 (m, 1H), 1.69 - 1.62 (m, 1H), 1.51 (d, J = 6.1 Hz, 3H), 1.06 (br dd, J = 3.0, 6.6 Hz, 3H), 1.03 - 1.00 (m, 3H), 0.96 (br s, 3H), 0.46 (s, 3H) ppm.

Example 15

(2S)-2-[8-[(2/f)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diaz aspiro[4.5]decan-2-yl]-A^-[(7S,13S)-

24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylp iperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(8-tert-butoxycarbonyl-l-oxo-2,8-diazaspiro[4.5]decan -2-yl)-3-methyl-butanoic acid (compound 15F) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J). Example 15 (56.8 mg) was obtained as a white solid. MS calc’d 1131.5 (MH ⁺ ), measured 1131.5 (MH ⁺ ). ' H NMR (400 MHz, Methanol-^) 6 = 8.86 (d, 7 = 7.2 Hz, 1H), 8.68 (d, 7 = 7.6 Hz, 1H), 8.50 (d, 7 = 2.8 Hz, 1H), 7.71 (s, 1H), 7.50 - 7.46 (m, 1H), 7.13 - 6.92 (m, 1H), 5.68 (br s, 1H), 5.20 - 5.13 (m, 1H), 4.41 (d, 7 = 14.1 Hz, 1H), 4.36 - 4.21 (m, 4H), 4.20 - 3.85 (m, 4H), 3.82 - 3.68 (m, 3H), 3.61 (d, 7 = 7.3 Hz, 4H), 3.46 (d, 7 = 14.4 Hz, 4H), 3.36 - 3.35 (m, 2H), 3.24 - 3.08 (m, 4H), 3.00 (s, 3H), 2.90 - 2.78 (m, 1H), 2.57 (d, 7 = 14.4 Hz, 1H), 2.28 - 2.17 (m, 2H), 2.08 (s, 2H), 1.96 ( d, 7 = 12.8 Hz, 2H), 1.88 - 1.77 (m, 2H), 1.76 - 1.52 (m, 4H), 1.45 (d, 7 = 6.0 Hz, 3H), 1.00 (d, 7 = 6.4 Hz, 3H), 0.97 (s, 3H), 0.92 - 0.84 (m, 3H), 0.44 (s, 3H) ppm.

The compound 15F was prepared according to the following scheme:

Step 1: Preparation of butyl Oa-methyl 4-allylpiperidine-l,4-dicarboxylate (compound 15B)

To a solution of Oi-tert-butyl O4- methyl piperidine- 1,4-dicarboxy late (compound 15A, 10.0 g, 41.1 mmol) in THF (100 mL) was added LDA (22.6 mL, 45.21 mmol) dropwise at -70 °C under nitrogen atmosphere. After being stirred for 0.5 h, the reaction mixture was added with allyl bromide (5.5 g, 45.21 mmol) at -70 °C, and then the reaction mixture was allowed to warm up to 15 °C and stirred for 2 hrs. After the reaction was completed, it was quenched with saturated NH4CI aqueous solution (300 mL) and the reaction mixture was extracted with EtOAc (100 mL, three times). The combined organic layer was washed with brine (200 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum. The residue was purified by silica gel chromatography (EtOAc in PE = 5 % to 30 %) to afford Oi-tert-butyl O4-methyl 4- allylpiperidine- 1,4-dicarboxy late (compound 15B, 10.8 g) as yellow oil. MS calc’d 284.2 (MH ⁺ ), measured 228.2 (M-C4Hs+H ⁺ ).

Step 2: Preparation of Oi-/('/7-butyl 04-methyl 4-(2-oxoethyl)piperidine-l,4- dicarboxylate (compound 15C)

To a mixture of Oi-terZ-butyl O4-methyl 4-allylpiperidine- 1,4-dicarboxy late (compound 15B, 6.0 g, 21.17 mmol) in 1,4-dioxane (200 mL) and water (20 mL) were added 2,6-Lutidine (4.9 mL, 42.35 mmol) and potassium osmate(VI)dihydrate (0.4 g, 1.06 mmol) in one portion at 0 °C. After being stirred at 0 °C for 15 minutes, the reaction mixture was added with sodium metaperiodate (18.1 g, 84.7 mmol) portion-wise at 0 °C and the resulting mixture was warmed to 20 °C and stirred for 2 hrs. The reaction was quenched with saturated Na2SOs aqueous solution (200 mL) and the reaction mixture was extracted with EtOAc (100 mL, three times). The combined organic layer was washed with brine (100 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to afford Oi-tert-butyl CL- methyl 4-(2-oxoethyl)piperidine-l,4- dicarboxylate (compound 15C, 6.0 g) as yellow oil. MS calc’d 286.2 (MH ⁺ ), measured 186.2 (M-C ₅ H ₈ O2+H ⁺ ).

Step 3: Preparation of Oi-/c/7- butyl (h-methyl 4-[2-[[(lS)-l-benzyloxycarbonyl-2- methyl-propyl]amino]ethyl]piperidine-l,4-dicarboxylate (compound 15D)

To a mixture of benzyl (2S)-2-amino-3-methyl-butanoate hydrogen chloride (2.2 g, 10.51 mmol) and Oi-tert-butyl CL-methyl 4-(2-oxoethyl)piperidine-l,4-dicarboxylate (compound 15C, 2.0 g, 7.01 mmol) in methanol (30 mL) was added zinc chloride (1.4 g, 10.51 mmol) at 0 °C. After being stirred at 0 °C for 0.5 h, the reaction mixture was added with sodium cyanoborohydride (0.9 g, 14.02 mmol). After being stirred at 20 °C for another 1 h, the reaction was quenched with saturated NH4CI aqueous solution (100 mL) and the reaction mixture was extracted with EtOAc (50 mL, three times). The combined organic layer was washed with brine (50 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE = 10 % to 30 %) to afford Oi-tert- butyl O4-methyl 4-[2-[[(lS)-l-benzyloxycarbonyl-2-methyl-propyl]amino]ethyl] piperidine-l ,4- dicarboxylate (compound 15D, 2.46 g) as yellow oil. MS calc’d 477.3 (MH ⁺ ), measured 477.3 (MH ⁺ ).

Step 4: Preparation of tert-butyl 2-[(lS)-l-benzyloxycarbonyl-2-methyl-propyl]-l-oxo- 2,8-diazaspiro[4.5]decane-8-carboxylate (compound 15E)

To a mixture of 1 -(tert-butyl) 4-methyl (S)-4-(2-((l-(benzyloxy)-3-methyl-l-oxobutan-2- yl)amino)ethyl)piperidine-l,4-dicarboxylate (compound 15D, 1.2 g, 2.52 mmol) in toluene (20 mL) were added DIEA (4.4 mL, 25.18 mmol) and DMAP (0.3 g, 2.52 mmol). The reaction mixture was heated to 100 °C and stirred for 30 hrs. After the reaction was completed, the reaction mixture was poured into water (30 mL) and extracted with EtOAc (30 mL, three times). The combined organic layer was washed with brine (30 mL), dried with anhydrous Na2SO4, filtered and concentrated under vacuum. The residue was purified by reversed phase column to afford tert-butyl 2-[(15)-l-benzyloxycarbonyl-2-methyl-propyl]-l-oxo-2,8- diazaspiro[4.5]decane-8-carboxylate (compound 15E, 586.0 mg) as yellow oil. MS calc’d 445.3 (MH ⁺ ), measured 467.4 (MNa ⁺ ).

Step 5: Preparation of (2.S)-2-(8-/er/-butoxycarbonyl-l-oxo-2,8-diazaspiro|4.5]deca n- 2-yl)-3-methyl-butanoic acid (compound 15F) To a solution of tert-butyl 2-[(15)-l-benzyloxycarbonyl-2-methyl-propyl]-l-oxo-2,8- diazaspiro[4.5]decane-8-carboxylate (compound 15E, 500.0 mg) in toluene (20 mL) was added wet palladium on activated carbon (50.0 mg, 10%wt). The mixture was degassed and purged with H2 for 3 times. The mixture was hydrogenated at 35 °C. The reaction solution was filtered and the filter cake was washed with DMF (100 mL). The combined filtrate was concentrated in vacuo to afford (2S)-2-(8-tert-butoxycarbonyl-l-oxo-2,8-diazaspiro[4.5]decan -2-yl)-3-methyl- butanoic acid (compound 15F, 144.0 mg) as a white solid. MS calc’d 355.2 (MH ⁺ ), measured 377.1 (MNa ⁺ ).

Example 16 and Example 18 (2S)-2-[(5S)-2-[(2/f)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-di azaspiro[4.5]decan-7-yl]-2V- [(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 16) and (2S)-2-[(5/f)-2-[(2/f)-2-chloro-2-fluoro-acetyl]-6-oxo- 2,7-diazaspiro[4.5]decan-7-yl]-^-[(7S,13S)-24-fluoro-(20M)-2 0-[2-[(lS)-l-methoxyethyl]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-diox o-21-(2,2,2-trifluoroethyl)-15- oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]-3-methyl-butanamide (Example 18)

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5] decan-7-yl]-3-methyl-butanoic acid (compound 16o 1) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J). Example 16 (14.0 mg) was obtained as a white solid. MS calc’d 1131.5 (MH ⁺ ), measured 1131.4 (MH ⁺ ). ^J H NMR (400MHz, Methanol-^4) 3 = 8.69 (dd, J = 2.0, 7.6 Hz, 1H), 8.50 (d, J = 2.8 Hz, 1H), 7.80 - 7.70 (m, 1H), 7.51 - 7.45 (m, 2H), 6.91

- 6.73 (m, 1H), 5.72 - 5.64 (m, 1H), 5.23 - 5.13 (m, 2H), 4.42 (d, J = 12.0 Hz, 1H), 4.27 - 4.20 (m, 2H), 4.10 - 4.00 (m, 3H), 3.93 - 3.82 (m, 2H), 3.79 - 3.69 (m, 3H), 3.66 - 3.56 (m, 3H), 3.52

- 3.43 (m, 4H), 3.36 - 3.34 (m, 4H), 3.19 - 3.12 (m, 2H), 3.00 (s, 3H), 2.87 - 2.79 (m, 1H), 2.60 - 2.53 (m, 1H), 2.52 - 2.41 (m, 1H), 2.39 - 2.18 (m, 3H), 2.07 - 1.81 (m, 8H), 1.68 - 1.61 (m, 1H), 1.45 (d, J = 6.0 Hz, 3H), 1.00 - 0.96 (m, 6H), 0.90 (d, J = 6.4 Hz, 3H), 0.47 - 0.41 (m, 3H) ppm.

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-[(5R)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5] decan-7-yl]-3-methyl-butanoic acid (compound 16o 2) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J). Example 18 (25.6 mg) was obtained as a white solid. MS calc’d 1131.4 (MH ⁺ ), measured 1131.5(MH ⁺ ). ' H NMR (400MHz, Methanol-^) 3 = 8.68 (d, J = 7.6 Hz, 1H), 8.50 (d, J = 2.8 Hz, 1H), 7.74 - 7.71 (m, 1H), 7.50 - 7.45 (m, 2H), 6.91 - 6.76 (m, 1H), 5.72 - 5.65 (m, 1H), 5.24 - 5.11 (m, 2H), 4.83 - 4.79 (m, 3H), 4.47 - 4.18 (m, 2H), 4.12

- 3.87 (m, 3H), 3.81 - 3.60 (m, 6H), 3.56 - 3.44 (m, 3H), 3.41 - 3.33 (m, 6H), 3.26 - 3.12 (m, 3H), 3.00 (s, 3H), 2.87 - 2.78 (m, 1H), 2.26 - 2.46 (m, 2H), 2.39 - 2.19 (m, 2H), 2.05 - 1.79 (m, 7H), 1.71 - 1.60 (m, 1H), 1.44 (d, J = 6.4 Hz, 3H), 1.02 - 0.95 (m, 6H), 0.90 (d, J = 6.4Hz, 3H), 0.44 (s, 3H) ppm.

The compound 16o 1 and compound 16o 2 was prepared according to the following scheme:

Step 1: Preparation of l-(tert-butyl) 3-methyl 3-(but-3-en-l-yl)pyrrolidine-l,3- dicarboxylate (compound 16c) To a solution of 1 -(tert-butyl) 3-methyl pyrrolidine- 1,3-dicarboxylate (compound 16a, 25.0 g, 109.04 mmol) in THF (300 mL) was added LDA (59.9 mL, 119.95 mmol) at -70 °C under nitrogen atmosphere. After being stirred for 0.5 h, the reaction mixture was added with 4-bromo- 1-butene (compound 16b, 16.1 g, 119.95 mmol). After being stirred at 20 °C for 2.5 hrs, the reaction was quenched with saturated NH4CI (100 mL) and the reaction mixture was extracted with EtOAc (150 mL, twice). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by silica gel column (EtOAc in PE = 0% ~ 5%) to afford 1 -(tert-butyl) 3-methyl 3-(but-3-en-l- yl)pyrrolidine-l,3-dicarboxylate (compound 16c, 13.2 g) as yellow oil. ^J H NMR (400MHz, Methanol-^) 6 = 5.83 - 5.73 (m, 1H), 5.05 - 4.93 (m, 2H), 3.84 (d, J = 10.8 Hz, 1H), 3.71 (s, 3H), 3.46 - 3.37 (m, 1H), 3.29 - 3.11 (m, 2H), 2.40 - 2.32 (m, 1H), 2.01 - 1.91 (m, 2H), 1.90 - 1.74 (m, 3H), 1.46 (d, J = 3.2 Hz, 9H) ppm.

Step 2: Preparation of methyl 3-(but-3-en-l-yl)pyrrolidine-3-carboxylate (compound 16d)

To a solution of 1 -(tert-butyl) 3-methyl 3-(but-3-en-l-yl)pyrrolidine-l,3-dicarboxylate (compound 16c ,13.2 g, 46.58 mmol) in 1,4-dioxane (50 mL) was added HC1 solution (50.0 mL, 4 M in dioxane). After being stirred at 20 °C for 0.5 h, the reaction mixture was concentrated under vacuum to afford methyl 3-(but-3-en-l-yl)pyrrolidine-3-carboxylate hydrochloride salt (compound 16d, 10.2 g) as yellow oil.

Step 3: Preparation of methyl 3-(but-3-en-l-yl)-l-tritylpyrrolidine-3-carboxylate (compound 16e)

To a solution of methyl 3-(but-3-en-l-yl)pyrrolidine-3-carboxylate hydrochloride salt (compound 16d, 3.0 g, 13.65 mmol) in ACN (70 mL) were added triphenylmethyl chloride (3.81 g, 13.65 mmol) and potassium carbonate (4.72 g, 34.14 mmol). The mixture was stirred at 20 °C for 12 hrs. After the reaction was completed, the mixture was poured into water (100 mL) and extracted with EtOAc (30 mL, three times). The combined organic layer was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by silica gel column (EtOAc in PE = 0% ~ 5%) to afford methyl 3- (but-3-en-l-yl)-l-tritylpyrrolidine-3-carboxylate (compound 16e, 1.3 g) as yellow oil. ' H NMR (400MHz, CDCh) d = 7.48 - 7.45 (m, 6H), 7.26 - 7.23 (m, 6H), 7.15 (t, J = 7.2Hz, 3H), 5.78 - 5.68 (m, 1H), 5.00 - 4.89 (m, 2H), 3.74 (s, 3H), 3.00 (d, J = 9.6Hz, 1H), 2.61 - 2.55 (m, 1H), 2.43 - 2.35 (m, 1H), 2.20 - 2.14 (m, 1H), 2.03 (s, 1H), 1.98 - 1.83 (m, 2H), 1.82 - 1.61 (m, 2H), 1.52 - 1.45 (m, 1H) ppm.

Step 4: Preparation of methyl 3-(3-oxopropyl)-l-tritylpyrrolidine-3-carboxylate (compound 16f)

To a solution of methyl 3-(but-3-en-l-yl)-l-tritylpyrrolidine-3-carboxylate (compound 16e, 660 mg, 1.55 mmol ) in THF (20 mL) and water (10 mL) were added potassium osmate (VI) (5 mg, 0.02 mmol) and sodium periodate (663 mg, 3.1 mmol). The reaction mixture was stirred at 20 °C for 2 hrs. After the reaction was completed, it was quenched with sat. NH4CI aqueous solution and the reaction mixture was extracted with EtOAc (50 mL, twice). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by silica gel column (EtOAc in PE = 0% ~ 5%) to afford methyl 3-(3-oxopropyl)-l-tritylpyrrolidine-3-carboxylate (compound 16f, 300 mg) as yellow oil. ^J H NMR (400MHz, CDCh) d = 9.75 - 9.69 (m, 1H), 7.52 - 7.48 (m, 6H), 7.29 (t, J = 4.0 Hz, 6H), 7.19 (t, J = 7.2Hz, 3H), 3.77 (s, 3H), 3.05 (d, J = 9.2 Hz, 1H), 2.67 - 2.57 (m, 1H), 2.47 - 2.31 (m, 3H), 2.29 - 2.21 (m, 1H), 2.04 - 1.87 (m, 2H), 1.62 - 1.43 (m, 2H) ppm.

Step 5: Preparation of methyl 3-(3-(((S)-l-(tert-butoxy)-3-methyl-l-oxobutan-2- yl)amino)propyl)-l-tritylpyrrolidine-3-carboxylate (compound 16h)

To a mixture of methyl 3-(3-oxopropyl)-l-tritylpyrrolidine-3-carboxylate (compound 16f ,550.0 mg, 1.29 mmol), H-VAL-OTBU HC1 (compound 16g, 296.7 mg, 1.42 mmol) and zinc chloride (192.8 mg, 1.42 mmol) in methanol (6 mL) was added sodium cyanoborohydride (88.9 mg, 1.42 mmol) at 0 °C. The mixture was stirred at 25 °C for 2 hrs. After the reaction was completed, water (30 mL) was added to the reaction mixture. The resulting mixture was concentrated under vacuum. The resulting suspension was diluted with water (40 mL) and extracted with EtOAc (30mL, twice). The combined organic layer was washed with brined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue which was purified by column chromatography (EtOAc in PE = 0% ~ 10%) to afford methyl 3-(3-(((5)- l-(tert-butoxy)-3-methyl-l-oxobutan-2-yl)amino)propyl)-l-tri tylpyrrolidine-3-carboxylate (compound 16h, 510 mg) as yellow oil. ' H NMR (400MHz, CDCh) d = 7.47 (d, J = 8.0 Hz, 6H), 7.28 - 7.27 (m, 1H), 7.27 - 7.23 (m, 5H), 7.18 - 7.13 (m, 3H), 3.78 - 3.73 (m, 3H), 3.05 (t, J = 8.4Hz, 1H), 2.79 (dd, J = 2.4, 6.0 Hz, 1H), 2.67 - 2.59 (m, 1H), 2.55 - 2.49 (m, 1H), 2.43 - 2.31 (m, 2H), 2.14 - 2.09 (m, 1H), 1.99 - 1.94 (m, 1H), 1.88 - 1.82 (m, 1H), 1.68 - 1.64 (m, 3H), 1.60 - 1.55 (m, 2H), 1.48 - 1.45 (m, 9H), 1.37 - 1.33 (m, 1H), 0.94 - 0.90 (m, 6H) ppm.

Step 6: Preparation of 3-(3-(((S)-l-(tert-butoxy)-3-methyl-l-oxobutan-2- yl)amino)propyl)-l-tritylpyrrolidine-3-carboxylic add (compound 16i)

To a solution of methyl 3-(3-(((S)-l-(tert-butoxy)-3-methyl-l-oxobutan-2- yl)amino)propyl)-l-tritylpyrrolidine-3-carboxylate (compound 16h, 510 mg, 0.8 mmol) in methanol (6 mL), water (0.6 mL) and THF (0.6 mL) was added lithium hydroxide monohydrate (336 mg, 8 mmol). The mixture was stirred at 60 °C for 12 hrs. After the reaction was completed, the mixture was diluted with water (20 mL), neutralized with 1 M HC1 aqueous and extracted with EtOAc (20 mL, twice). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford 3-(3-(((S)-l-(tert- butoxy)-3-methyl- l-oxobutan-2-yl)amino)propyl)- l-tritylpyrrolidine-3-carboxylic acid (compound 16i, 440 mg) as white gum. ^J H NMR (400MHz, CDCE) 8 = 7.46 (d, J = 8.0 Hz, 6H), 7.26 - 7.19 (m, 6H), 7.16 - 7.10 (m, 3H), 3.10 - 2.86 (m, 4H), 2.54 - 2.45 (m, 2H), 2.37 - 2.22 (m, 2H), 2.18 - 2.10 (m, 1H), 2.04 - 1.97 (m, 1H), 1.68 - 1.54 (m, 2H), 1.49 - 1.44 (m, 9H), 1.41 - 1.35 (m, 2H), 0.99 - 0.89 (m, 6H) ppm.

Step 7: Preparation of tert-butyl (2S)-3-methyl-2-(6-oxo-2-trityl-2,7- diazaspiro[4.5]decan-7-yl)butanoate (compound 16j)

To a solution of 3-(3-(((S)-l-(tert-butoxy)-3-methyl-l-oxobutan-2-yl)amino)pr opyl)-l- tritylpyrrolidine-3-carboxylic acid (compound 16i, 440 mg, 0.77 mmol) in DMF (5 mL) was added DIEA (0.67 mL, 3.85 mmol) and COMU (594 mg, 1.39 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (20 mL, twice). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by silica gel column (EtOAc in PE = 0% ~ 10%) to afford tert-butyl (2S)-3-methyl-2-(6-oxo-2-trityl-2,7-diazaspiro[4.5]decan-7-y l)butanoate (compound 16j, 420 mg) as yellow oil. MS calc’d 553.3 (MH ⁺ ), measured 553.2 (MH ⁺ ).

Step 8: Preparation of (2S)-3-methyl-2-(6-oxo-2,7-diazaspiro[4.5]decan-7-yl)butanoi c add (compound 16k)

To a solution of tert-butyl (2S)-3-methyl-2-(6-oxo-2-trityl-2,7-diazaspiro[4.5]decan-7- yl)butanoate (compound 16j, 550 mg, 1 mmol) in DCM (3 mL) was added TFA (3.0 mL). The mixture was stirred at 20 °C for 12 hrs. After the reaction was completed, the mixture was concentrated under vacuum to give a residue, which was dissolved in water (10 mL) and the aqueous phase was washed with EtOAc (20 mL, twice). The aqueous phase was concentrated under vacuum to afford (2S)-3-methyl-2-(6-oxo-2,7-diazaspiro[4.5]decan-7-yl)butanoi c acid in TFA salt form(compound 16k, 365 mg) as colorless oil. MS calc’d 255.2 (MH ⁺ ), measured 255.2 (MH ⁺ )

Step 9: Preparation of (2.S')-2-(2-(/cr/-butoxycarbonyl)-6-oxo-2,7- diazaspiro[4.5]decan-7-yl)-3-methylbutanoic acid (compound 161).

To a solution of (2S)-3-methyl-2-(6-oxo-2,7-diazaspiro[4.5]decan-7-yl)butanoi c acid in TFA salt form (compound 16k, 365 mg, 0.99 mmol) in THF (10 mL) and water (10 mL) were added sodium carbonate (210 mg, 1.98 mmol) and di-t-butyldicarbonate (259 mg, 1.19 mmol). The mixture was stirred at 20 °C for 2 hrs. After the reaction was completed, the mixture was neutralized with 1 M HC1 aq. and the reaction mixture was extracted with EtOAc (20 mL, twice). The combined organic layer was washed with brine, dried over sodium sulfate and concentrated under vacuum to afford (2S)-2-(2-(tert-butoxycarbonyl)-6-oxo-2,7-diazaspiro[4.5]dec an-7-yl)-3- methylbutanoic acid (compound 161, 350 mg) as a white solid.

Step 10: Preparation of tert-butyl (5S)-7-[(lS)-l-tert-butoxycarbonyl-2-methyl- propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylate (compound 16m 1) and tert-butyl (5/f)-7-[(lS)-l-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2 ,7-diazaspiro[4.5]decane-2- carboxylate (compound 16m 2)

Tert-butyl 7-[(15)-l-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7- diazaspiro[4.5]decane-2-carboxylate (compound 161, 680.0 mg, 1.66 mmol) was separated by SFC (regis (S,S)whelk-Ol(250 mm, twice5mm x 10 um), mobile phase: (0.1%NH3H2O) IPA, B%: 15% -15%, 4.4 min) to afford tert-butyl (5S)-7-[(lS)-l-tert-butoxycarbonyl-2-methyl- propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylate (compound 16m 1, 245.0 mg, faster eluted) as yellow oil and tert-butyl (57?)-7-[(15)-l-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo- 2,7-diazaspiro[4.5]decane-2-carboxylate (compound 16m 2, 223.0 mg, slower eluted) as yellow oil. MS calc’d 411.3 (MH ⁺ ), measured 411.3 (MH ⁺ ).

Step 11: Preparation of (2S)-3-methyl-2-[(5S)-6-oxo-2,7-diazaspiro[4.5]decan-7- yl]butanoic acid (compound 16n 1)

To a solution of tert-butyl (5S)-7-[(lS)-l-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2, 7- diazaspiro[4.5]decane-2-carboxylate (compound 16m 1, 245.0 mg, 0.6 mmol) in DCM (1 mL) was added TFA (1.0 mL, 12.98 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the mixture was concentrated under vacuum to afford (2S)-3-methyl-2- [(5S)-6-oxo-2,7-diazaspiro[4.5]decan-7-yl]butanoic acid (compound 16n 1, 219.0 mg, TFA salt) as colorless liquid, which was used directly in the next step. MS calc’d 255.2 (MH ⁺ ), measured 255.2 (MH ⁺ ).

Step 12: Preparation of (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7- diazaspiro[4.5]decan-7-yl]-3-methyl-butanoic acid (compound 16 ol)

To a solution of (2S)-3-methyl-2-[(5S)-6-oxo-2,7-diazaspiro[4.5]decan-7-yl]bu tanoic acid (compound 16n 1, 219.0 mg, 0.59 mmol) in THF (2 mL) and water (2 mL) was added sodium carbonate (189.05 mg, 1.78 mmol) and di-t-butyldicarbonate (155.71 mg, 0.71 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the mixture was filtered and the filtrate was concentrated under vacuum to give a residue, which was purified by reversed phase flash column to afford (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5] decan- 7-yl]-3-methyl-butanoic acid (compound 16o 1, 200.0 mg) as a white solid. MS calc’d 353.2 (MH’), measured 353.0 (MH’).

Step 13: Preparation of (2S)-3-methyl-2-[(5/f)-6-oxo-2,7-diazaspiro[4.5]decan-7- yl]butanoic add (compound 16n 2)

To a solution of tert-butyl (57?)-7-[(15)-l-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2 ,7- diazaspiro[4.5]decane-2-carboxylate (compound 16m 2, 223.0 mg, 0.54 mmol) in DCM (1.5 mL) was added TFA (1.5 mL, 19.47 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the mixture was concentrated under vacuum to afford (2S)-3-methyl-2- [(57?)-6-oxo-2,7-diazaspiro[4.5]decan-7-yl]butanoic acid (compound 16n 2, 200.0 mg, TFA salt) as colorless liquid, which was used directly in the next step. MS calc’d 255.2 (MH ⁺ ), measured 255.1 (MH ⁺ ).

Step 14: Preparation of (2.S )-2-|(5/?)-2-/er/-butoxycarbonyl-6-oxo-2,7- diazaspiro[4.5]decan-7-yl]-3-methyl-butanoic acid (compound 16o 2)

To a solution of (2S)-3-methyl-2-[(57?)-6-oxo-2,7-diazaspiro[4.5]decan-7-yl]b utanoic acid (compound 16n 2, 200.0 mg, 0.54 mmol) in THF (2 mL) and water (2 mL) was added sodium carbonate (115.1 mg, 1.09 mmol) and di-t-butyldicarbonate (142.2 mg, 0.65 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the mixture was filtered and filtrate was concentrated under vacuum to give a residue, which was purified by reversed phase flash column to afford (2S)-2-[(57?)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5 ]decan-7-yl]- 3-methyl-butanoic acid (compound 16 o2, 150.0 mg) as a white solid. MS calc’d 355.2 (MH ⁺ ), measured 377.2 (MNa ⁺ ).

Example 20

(2.S)-2-|9-|(2/?)-2-chloro-2-fluoro-acetyl|-l -oxo-2, 9-diazaspiro|5.5]undecan-2-yl|-V- [(7S,13S)-25-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4- methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2-[(15')-l-methoxyethy l]-5-(4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-t hia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E) instead of (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')-l-methoxyethy l]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D). Example 20 (6.1 mg) was obtained as a white solid. MS calc’d 1145.5 (MH ⁺ ), measured 1145.5 (MH ⁺ ). ^J H NMR (400MHz, Methanol-^) d = 8.52 (s, 1H), 7.61 - 7.45 (m, 4H), 7.09 - 6.91 (m, 1H), 5.97 - 5.84 (m, 1H), 5.24 - 5.06 (m, 3H), 4.69 - 4.56 (m, 0.5H), 4.44 - 4.33 (m, 0.5H), 4.20 - 3.96 (m, 3H), 3.90 - 3.40 (m, 10H), 3.27 - 3.10 (m, 12H), 3.00 (s, 3H), 2.95 - 2.75 (m, 1H), 2.71 - 2.53 (m, 1H), 2.25 - 2.04 (m, 2H), 2.00 - 1.79 (m, 4H), 1.72 - 1.53 (m, 3H), 1.44 (d, J = 6.0 Hz, 3H), 1.37 - 1.19 (m, 2H), 0.94 (d, J = 6.0 Hz, 3H), 0.89 - 0.73(m, 7H), 0.71 - 0.53 (m, 3H) ppm.

Example 22 and Example 23

(2S)-2-[(5/f)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7 -diazaspiro[4.5]decan-7-yl]-2V-

[(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxy ethyl]-5-(4-methylpiperazin-l-yl)- 3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,2 8- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 22) and (2S)-2-[(5S)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo- 2,7-diazaspiro[4.5]decan-7-yl]-^-[(7S,13S)-21-ethyl-24-fluor o-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ^2,5 .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]-3-methyl-butanamide (Example 23) The title compounds were prepared in analogy to the preparation of Example 6 by using (2S)-2-chloro-2-fluoro-acetic acid, (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7- diazaspiro[4.5]decan-7-yl]-3-methyl-butanoic acid (compound 16o 1) and (7S,13S)-7-amino-21- ethyl-24-fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-(4-meth ylpiperazin-l-yl)-3-pyridyl]-

25 913 22226 17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17 .5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (27?)-2-chloro-2- fluoro-acetic acid, (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2-yl)-3- methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 2 5 9 13 222 26 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 22 (4.1 mg) was obtained as a white solid. MS calc’d 1077 (MH ⁺ ), measured 1077 (MH ⁺ ). ¹ HNMR (400 MHz, DMSO-<7 ₆ ) 6 9.98 - 10.20 (m, 1 H), 8.42 - 8.62 (m, 2 H), 8.10 - 8.34 (m, 1 H), 7.71 (d, J = 2.50 Hz, 1 H), 7.57 (d, 7=13.01 Hz, 1 H), 7.39 (br s, 1 H), 7.15 (d, J = 10.01 Hz, 1 H), 7.00 - 7.07 (m, 1 H), 5.41 - 5.54 (m, 1 H) , 5.18 (br dd, J = 19.51, 11.88 Hz, 1 H), 4.81 (br dd, J = 12.88, 11.38 Hz, 1 H), 4.23 - 4.28 (m, 2 H), 4.13 (br s, 5 H), 3.57 (br s, 3 H), 3.34 - 3.34 (m, 3 H), 2.82 - 2.94 (m, 3 H), 2.66 - 2.81 (m, 7 H), 2.41 (br s, 1 H), 2.31 - 2.34 (m, 3 H), 2.21 (br d, 7 = 6.88 Hz, 1 H), 1.69 - 1.88 (m, 7 H), 1.50 (br s, 1 H), 1.33 (d, 7 = 6.13 Hz, 3 H), 1.18 - 1.26 (m, 3 H), 0.84 - 0.98 (m, 11 H), 0.80 (br t, 7 = 7.32 Hz, 3 H), 0.35 (s, 3 H) ppm.

Example 23 was prepared in analogy to the preparation of Example 6 by using (25)-2- chloro-2-fluoro-acetic acid, (2S)-2-[(5R)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5] decan- 7-yl]-3-methyl-butanoic acid (compound 16o 2) and (7S,13S)-7-amino-21-ethyl-24-fluoro- (20A7)-20-[2-[(15)- l-methoxyethyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl]- 17, 17-dimethyl- 15- 2 5 9 13 22 26 oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (2R)-2-chloro-2- fluoro-acetic acid, (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2-yl)-3- methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 2 5 9 13 22 26 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 23 (2.2 mg) was obtained as a white solid. MS calc’d 1077 (MH ⁺ ), measured 1077 (MH ⁺ ). ¹ H NMR (400 MHz, METHANOL-7 ₄ ) 8 ppm 8.66 (d, 7 = 7.50 Hz, 1 H), 8.47 (d, 7 = 2.88 Hz, 1 H), 7.66 (br d, 7 = 7.13 Hz, 1 H), 7.45 (d, 7 = 2.75 Hz, 1 H), 7.33 - 7.36 (m, 1 H), 6.73 - 6.94 (m, 1 H), 5.32 - 5.38 (m, 2 H), 4.37 - 4.53 (m, 2 H), 4.09 - 4.30 (m, 5 H), 3.68 - 3.81 (m, 4 H), 3.61 (br d, 7 = 7.00 Hz, 4 H), 3.04 (q, 7 = 7.75 Hz, 4 H), 2.94 (br s, 3 H), 2.77 - 2.87 (m, 2 H), 2.54 - 2.66 (m, 2 H), 2.16 - 2.22 (m, 3 H), 2.01 - 2.05 (m, 3 H), 1.88 (br s, 2 H), 1.63 (dt, 7 = 15.26, 4.31 Hz, 8 H), 1.43 (d, 7 = 6.25 Hz, 3 H), 0.89 - 1.00 (m, 15 H), 0.49 (br d, 7 = 2.63 Hz, 3 H).

Example 24

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,1 3S)-24-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-di oxo-21-(2,2,2-

25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolid ine- 3-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l-methoxyethy l]-5-morpholino-3-pyridyl]- 17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21, 27,28-tetrazapentacyclo- [17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate I) instead of (75,13S)-7-amino-25-fhioro-(20M)-20-[2-[(15)-l-methoxyethyl] -5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate E). Example 24 (18.9 mg) was obtained as a white solid. MS calc’d 1092.4 (MH ⁺ ), measured 1092.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.79 - 8.66 (m, 1H), 8.60 - 8.35 (m, 1H), 7.81 - 7.67 (m, 2H), 7.57 - 7.42 (m, 1H), 6.94 - 6.72 (m, 1H), 5.76 - 5.61 (m, 1H), 5.33 - 5.10 (m, 1H), 4.80 - 4.60 (m, 2H), 4.43 - 4.10 (m, 3H), 3.88 - 3.52 (m, 12H), 3.43 - 3.36 (m, 8H), 3.11 - 2.99 (m, 4H), 2.85 - 2.81 (m, 1H), 2.68 - 2.64 (m, 1H), 2.34 - 2.21 (m, 4H), 1.95 - 1.61 (m, 3H), 1.48 - 1.47 (m, 3H), 1.07 - 0.81 (m, 9H), 0.60 - 0.42 (m, 3H) ppm.

Example 25 (2S)-2-[2-[(2/f)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazasp iro[3.4]octan-6-yl]-A^-[(7S,13S)- 24-fluoro-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l -yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(8-tert-butoxycarbonyl-l-oxo-2,8-diazaspiro[4.5]decan -2-yl)-3-methyl-butanoic acid (compound 25H) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J). Example 25 (20.8 mg) was obtained as a white solid. MS calc’d 1103.4 (MH ⁺ ), measured 1103.7 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.96 (d, J = 8.4 Hz, 1H), 8.68 (d, J = 7.6 Hz, 1H), 8.50 (d, J = 2.8 Hz, 1H), 7.72 (dd, J = 2.4, 17.6 Hz, 1H), 7.54 - 7.41 (m, 2H), 6.80 - 6.61 (m, 1H), 5.72 - 5.60 (m, 1H), 5.21 - 5.14 (m, 1H), 4.65 - 4.55 (m, 1H), 4.45 - 4.38 (m, 2H), 4.38 - 4.28 (m, 2H), 4.26 - 4.19 (m, 2H), 4.16 - 4.00 (m, 3H), 3.81 - 3.67 (m, 3H), 3.66 - 3.53 (m, 3H), 3.51 - 3.39 (m, 4H), 3.35 (s, 3H), 3.25 - 3.10 (m, 3H), 3.00 (s, 3H), 2.89 - 2.78 (m, 1H), 2.60 - 2.52 (m, 1H), 2.47 - 2.36 (m, 2H), 2.28 - 2.19 (m, 2H), 1.99 - 1.92 (m, 1H), 1.89 - 1.78 (m, 1H), 1.70 - 1.59 (m, 1H), 1.44 (d, 7 = 6.0 Hz, 3H), 1.01 (dd, 7 = 3.6, 6.4 Hz, 3H), 0.97 (s, 3H), 0.92 - 0.86 (m, 3H), 0.43 (s, 3H) ppm.

The compound 25H was prepared according to the following scheme:

25G 25H

Step 1: tert-butyl 3-allyl-3-cyano-azetidine-l-carboxylate (compound 25B)

To a solution of tert-butyl 3-cyanoazetidine-l-carboxylate (compound 25A, 10.0 g, 54.88 mmol) in THF (100 mL) was added LDA (60.3 mL, 60.37 mmol) dropwise at -70 °C under nitrogen atmosphere. After being stirred at -70 °C for 0.5 h, the reaction mixture was added with allyl bromide (7.9 g, 65.85 mmol). The mixture was stirred at 16 °C for another 1 h, then concentrated in vacuo and the residue was purified by column chromatography (EtOAc in PE: 3%- 10%) to afford tert-butyl 3-allyl-3-cyano-azetidine-l-carboxylate (compound 25B, 12.5 g) as light yellow oil. ^J H NMR (400 MHz, CHLOROFORM-<7) 8 5.90 - 5.70 (m, 1 H), 5.32 (s, 1 H), 5.28 (d, J = 5.2 Hz, 1 H), 4.23 (d, J = 8.8 Hz, 2 H), 3.86 (d, J = 8.8 Hz, 2 H), 2.63 (d, J = 7.2 Hz, 2 H), 1.45 (s, 9 H) ppm.

Step 2: 3-allyl- l-tert-butoxycarbonyl-azetidine-3-carboxylic add (compound 25C)

A mixture of tert-butyl 3-allyl-3-cyano-azetidine-l -carboxylate (compound 25B, 12.5 g, 56.24 mmol) and potassium hydroxide (12.6 g, 224.94 mmol) in ethanol (60 mL) and water (60 mL) was stirred at 100 °C for 16 hrs. After the reaction was completed, the pH of the reaction mixture was acidified by HC1 (I M) aq. to pH=3. The reaction mixture was extracted with EtOAc (120 mL, three times). The combined organic layer was washed with brine (300 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford 3-allyl- 1 -tert- butoxycarbonyl- azetidine-3-carboxylic acid (compound 25C, 14.0 g) as a white solid. MS calc’d 241.1 (MH ⁺ ), measured 186.2 (M-C4H ₈ +H ⁺ ).

Step 3: Oi-tert-butyl Ov methyl 3-allylazetidine-l,3-dicarboxylate (compound 25D)

To a mixture of 3-allyl-l-tert-butoxycarbonyl-azetidine-3-carboxylic acid (compound 25C, 8.0 g, 33.16 mmol) in DMF (80 mL) was added potassium carbonate (13.7 g, 99.47 mmol) followed by iodomethane (9.4 g, 66.31 mmol). After being stirred at 20 °C for 2 hrs, the reaction mixture was poured into water (400 mL) and the resulting mixture was extracted with EtOAc (100 mL, three times). The combined organic layer was washed with brine (300 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford Oi-tert-butyl CL-mcthyl 3-allylazetidine- 1,3-dicarboxylate (compound 25D, 7.3 g) as yellow oil. ^J H NMR (400 MHz, CHLOROFORM-<7) 8 5.80 - 5.60 (m, 1 H), 5.19 - 5.14 (m, 1 H), 5.12 (s, 1 H), 4.16 (d, J = 8.8 Hz, 2 H), 3.82 - 3.69 (m, 5 H), 2.63 (d, J = 7.2 Hz, 2 H), 1.44 (s, 9 H) ppm.

Step 4: Oi-/(77-butyl (h- methyl 3-(2-oxoethyl)azetidine-l,3-dicarboxylate (compound 25E)

To a mixture of Oi-tert-butyl Ch-mcthyl 3-allylazetidine-l,3-dicarboxylate (compound 25D, 2 g, 7.83 mmol) in 1,4-dioxane (20 mL) and water (20 mL) was added 2,6-Lutidine (1.8 mL, 15.67 mmol) and KaOsCL (144.3 mg, 0.39 mmol). After being stirred at 16 °C for 15 minutes, the reaction mixture was added with sodium metaperiodate (6702.2 mg, 31.33 mmol) and stirred for another 1 h, then poured into water (120 mL) and the resulting mixture was extracted with EtOAc (40 mL, three times). The combined organic layer was washed with sat. Na2SOs (60 mL), brine (60 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford Oi-tert-butyl Os-methyl 3-(2-oxoethyl)azetidine-l,3-dicarboxylate (compound 25E, 2.8 g) as yellow oil. MS calc’d 257.1 (MH ⁺ ), measured 202.1, (M-C ₄ H ₈ +H ⁺ ).

Step 5: Oi-tert-butyl O3-methyl 3-[2-[[(lS)-l-benzyloxycarbonyl-2-methyl- propyl]amino]ethyl]azetidine-l,3-dicarboxylate (compound 25F)

To a mixture of L-valine benzyl ester hydrochloride (3.2 g, 13.06 mmol) and Oi-tert-butyl O3-methyl 3-(2-oxoethyl)azetidine-l,3-dicarboxylate (compound 25E, 2.8 g, 10.88 mmol) in methanol (50 mL) was added zinc chloride (1.8 g, 13.06 mmol). After being stirred at 16 °C for 0.5 h, the reaction mixture was added with sodium cyanoborohydride (1.4 g, 21.77 mmol) and the resulting mixture was stirred for another 1 h. The reaction mixture was concentrated in vacuo and the residue was purified by column chromatography (EtOAc in PE: 30% - 50%) to afford Oi-tert-butyl Os-methyl 3-[2-[[(lS)-l-benzyloxycarbonyl-2-methyl- propyl]amino]ethyl]azetidine-l,3-dicarboxylate (compound 25F, 2.2 g,) as colorless oil. MS calc’d 449.3 (MH ⁺ ), measured 449.3 (MH ⁺ ).

Step 6: tert-butyl 6-[(lS)-l-benzyloxycarbonyl-2-methyl-propyl]-5-oxo-2,6- diazaspiro[3.4]octane-2-carboxylate (compound 25G)

To a mixture of 4-dimethylaminopyridine (544.7 mg, 4.46 mmol), DIEA (7.7 mL, 44.59 mmol) and Oi -/er/- butyl Ch- methyl 3-[2-[[(15)-l-benzyloxycarbonyl-2-methyl- propyl]amino]ethyl]azetidine-l,3-dicarboxylate (compound 25F, 2.0 g, 4.46 mmol) in toluene (2 mL) was stirred at 100 °C for 20 hrs. After the reaction was completed, the reaction mixture was concentrated in vacuo and the residue was dissolved in EtOAc (60 mL). The resulting solution was washed with HC1 aq. (1 M, 60 mL), then brine (60 mL), dried over Na2SO4, filtered. The filtrate was concentrated in vacuo to afford tert-butyl 6-[(15)-l-benzyloxycarbonyl-2-methyl- propyl]-5-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (compound 25G, 1.5 g) as a white solid. MS calc’d 417.2 (MH ⁺ ), measured 361.2 (M-C ₄ H ₈ +H ⁺ ).

Step 7: (2S)-2-(2-tert- butoxy carbonyl-5-oxo-2,6-diazaspiro[3.4]octan-6-yl)-3-methyl- butanoic acid (compound 25H)

A mixture of tert-butyl 6-[(15)-l-benzyloxycarbonyl-2-methyl-propyl]-5-oxo-2,6- diazaspiro[3.4]octane-2-carboxylate (compound 25G, 1.5 g, 3.6 mmol) and Pd on activated carbon (150.0 mg) in methanol (20 mL) was stirred under H2 balloon at 16 °C for 2 hrs. The reaction mixture was filtered and the filtrate was concentrated in vacuo to afford (25)-2-(2-tert- butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octan-6-yl)-3-methyl -butanoic acid (compound 25H, 920.0 mg) as a white solid. MS calc’d 327.2 (MH ⁺ ), measured 271.2 (M-C4H ₈ +H ⁺ ).

Example 26

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,1 3S)-24-fluoro-(20M)-20-[2-[(lS)-l- methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-l-yl]-3-p yridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbanioyl]-2-niethyl-propyl]-\-niethyl-pyrrolidine-3-car boxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (75,135)-7-amino-24-fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-21- (2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate J) instead of (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E). Example 26 (14.6 mg) was obtained as a white solid. MS calc’d 1173.4 (MH ⁺ ), measured 1173.7 (MH ⁺ ). ^J H NMR (400 MHz, METHANOL-^) 6 = 8.72 - 8.68 (m, 1H), 8.42 (d, J = 2.4 Hz, 1H), 7.77 - 7.68 m, 1H), 7.57 - 7.45 (m, 2H), 6.93 - 6.76 (m, 1H), 5.81- 5.63 (m, 1H), 5.27 - 5.11 (m, 1H), 4.96 - 4.93 (m, 1H), 4.88 - 4.79 (m, 2H), 4.47 - 4.38 (m, 1H), 4.28 - 4.18 (m, 2H), 3.88 - 3.53 (m, 7H), 3.47 -

3.34 (m, 8H), 3.30 - 3.24 (m, 1H), 3.23 - 3.07 (m, 6H), 2.92 - 2.80 (m, 5H), 2.70 - 2.56 (m, 1H),

2.40 - 2.06 (m, 4H), 2.00 - 1.91 (m, 1H), 1.88 - 1.74 (m, 1H), 1.69 - 1.57 (m, 1H), 1.49 - 1.41 (m,

3H), 1.01 - 0.97 (m, 5H), 0.92 - 0.82 (m, 3H), 0.53 - 0.42 (m, 3H) ppm.

Example 27 (3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(8S,14S) -26-fluoro-4-hydroxy-(21M)-21- [2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridy l]-18,18-dimethyl-9,15-dioxo- 22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacydo[18 .5.2.1 ² ’ ⁶ .l ¹⁰ ’ ¹⁴ .0 ²³ ’ ²⁷ ]nonacosa- l(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methy l-propyl]-2V-methyl- pyrrolidine-3-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (8S,14S)-8-amino-26-fluoro-4-hydroxy-(21A7)-21-[2-[(lS)-l-me thoxyethyl]-5-(4- methylpiperazin- l-yl)-3-pyridyl]- 18, 18-dimethyl-22-(2, 2, 2-trifluoroethyl)- 16-oxa- 10,22,28- triazapentacyc lo[ 18.5.2.1 ² ’ ⁶ . l ^{10 14} .0 ²³ ’ ²⁷ ]nonacosa- 1(26), 2, 4, 6(29), 20, 23(27), 24-heptaene-9, 15- dione (Intermediate K) instead of (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E). Example 27 (5.2 mg) was obtained as a white solid. MS calc’d 1114.5 (MEE), measured 1114.6 (MH ⁺ ), ^J H NMR (400 MHz, METHANOL-^) 6: 8.50 (d, J = 2.8 Hz, 1 H), 8.21- 8.10 (m, 1H), 7.51 (s, 1H), 7.47 (d, J = 8.0 Hz, 1H), 7.42 - 7.36 (m, 1H), 7.14 - 7.06 (m, 1H), 6.89 - 6.86 (m, 1H), 5.34 (t, J = 4.4 Hz, 1H), 4.69 (d, J = 10.8 Hz, 1H), 4.43 - 4.34 (m, 0.5H), 4.27 - 4.17 (m, 0.5H), 4.14 - 3.95 (m, 1H), 3.79 - 3.56 (m, 6H), 3.23 (s, 2H), 3.02 (d, J = 14 Hz, 6H), 2.91 - 2.78 (m, 2H), 2.78 - 2.56 (m, 2H), 2.19 (t, 7=7.6 Hz, 2H), 2.06 - 2.01 (m, 3H), 1.88 - 1.77 (m, 1H), 1.64 - 1.56 (m, 1H), 1.50 - 1.46 (m, 3H), 1.44 (d, 7 = 1.6 Hz, 1H), 1.34 - 1.29 (m, 10H), 1.18 (t, 7 = 6.8 Hz, 2H), 0.97 - 0.87 (m, 8H), 0.86 - 0.82 (m, 3H), 0.41 (s, 3H) ppm.

Example 28 (2S)-2-[8-[(2/f)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diazasp iro[4.5]decan-2-yl]-A^-[(7S,13S)- 21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-m ethylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(8-tert-butoxycarbonyl-l-oxo-2,8-diazaspiro[4.5]decan -2-yl)-3-methyl-butanoic acid (compound 15F) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]- 5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa- 4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 28 (11.7 mg) was obtained as a white solid. MS calc’d 1077.5 (MEE), measured 1077.5 (MH ⁺ ). ^J H NMR (400 MHz, METHANOL-^) 6 8.64 (d, J = 7.7 Hz, 1H), 8.43 (d, J = 1.7 Hz, 1H), 7.66 (s, 1H), 7.43 (d, J = 2.2 Hz, 1H), 7.30 (d, J = 12.3 Hz, 1H), 7.13 - 6.92 (m, 1H), 5.69 - 5.53 (m, 2H), 5.51 - 5.36 (m, 1H), 4.58 (s, 11H), 4.54 - 4.23 (m, 1H), 4.22 - 3.80 (m, 1H), 3.78 - 3.46 (m, 3H), 2.75 - 2.59 (m, 5H), 2.44 - 2.30 (m, 5H), 2.29 - 2.19 (m, 2H), 2.18 - 2.05 (m, 3H), 2.03 - 1.74 (m, 6H), 1.70 - 1.55 (m, 3H), 1.30 - 1.15 (m, 7H), 1.03 - 0.94 (m, 7H), 0.90 (dd, 7 = 2.6, 6.5 Hz, 3H), 0.61 (s, 3H) ppm.

Example 29

(2S)-2-[2-[(2/f)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-2V-[(7S,13S)-

21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-( 4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octan -6-yl)-3-methyl-butanoic acid (compound 25H) and (7S,13S)-7-amino-21-ethyl-24-fhioro-(20Af)-20-[2-[(lS)-l-met hoxyethyl]- 5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa- 4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 29 (63.2 mg) was obtained as a yellow solid. MS calc’d 1049.5 (MEE), measured 1049.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.76 - 8.63 (m, 1H), 8.58 - 8.45 (m, 1H), 7.78 - 7.56 (m, 2H), 7.44 - 7.30 (m, 1H), 6.82 - 6.59 (m, 1H), 5.83 - 5.67 (m, 1H), 4.65 - 4.53 (m, 1H), 4.50 - 3.99 (m, 11H), 3.88 - 3.67 (m, 3H), 3.61 - 3.47 (m, 3H), 3.47 - 3.42 (m, 3H), 3.41 - 3.35 (m, 4H), 3.32 - 3.22 (m, 2H), 3.11 - 3.04 (m, 1H), 3.01 (s, 3H), 2.90 - 2.78 (m, 1H), 2.69 - 2.60 (m, 1H), 2.45 - 2.35 (m, 2H), 2.31 - 2.16 (m, 2H), 2.03 - 1.92 (m, 1H), 1.90 - 1.75 (m, 1H), 1.72 - 1.58 (m, 1H), 1.52 - 1.41 (m, 3H), 1.08 - 0.98 (m, 6H), 0.96 (s, 3H), 0.89 (dd, J = 2.8, 6.4 Hz, 3H), 0.62 - 0.43 (m, 3H) ppm.

Example 30 and Example 31

(2S)-2-[(5/?)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2, 7-diazaspiro[4.4]nonan-2-yl]-2V-

[(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxy ethyl]-5-(4-methylpiperazin-l-yl)- 3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,2 8- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 30)

(Example 30)

(2S)-2-[(5S)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-2V-

[(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxy ethyl]-5-(4-methylpiperazin-l-yl)- 3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,2 8- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 31) (Example 31)

Example 30 was prepared in analogy to the preparation of Example 4 by using (75,135)-7- armno-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]- 5-(4-methylpiperazin-l-yl)-3- pyridyl]- 17, 17-dimethyl-15-oxa-4-thia-9, 21,27, 28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (7S,13S)-7-amino-24-fhioro-(20M)-20-[2-[(15)-l-methoxyethyl] -5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D). Example 30 (42.4 mg) was obtained as a yellow solid. MS calc’d 1063.5 (MH ⁺ ), measured 1063.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 8.72 - 8.65 (m, 1H), 8.56 - 8.45 (m, 1H), 7.78 - 7.73 (m, 0.5H), 7.70 - 7.64 (m, 0.5H), 7.63 - 7.57 (m, 1H), 7.41 - 7.31 (m, 1H), 6.95 - 6.71 (m, 1H), 5.82 - 5.67 (m, 1H), 4.51 - 4.02 (m, 8H), 3.99 - 3.90 (m, 1H), 3.88 - 3.70 (m, 5H), 3.69 - 3.56 (m, 4H), 3.52 - 3.43 (m, 3H), 3.37 (d, J = 6.4 Hz, 4H), 3.30 - 3.23 (m, 2H), 3.13 - 3.05 (m, 1H), 3.01 (s, 3H), 2.88 - 2.79 (m, 1H), 2.69 - 2.57 (m, 1H), 2.36 - 2.18 (m, 3H), 2.18 - 2.04 (m, 3H), 2.01 - 1.96 (m, 1H), 1.88 - 1.76 (m, 1H), 1.71 - 1.59 (m, 1H), 1.46 (d, J = 6.0 Hz, 3H), 1.05 - 0.94 (m, 9H), 0.94 - 0.89 (d, J = 6.6 Hz, 3H), 0.58 - 0.41 (m, 3H) ppm.

Example 31 was prepared in analogy to the preparation of Example 4 by using (25)-2- [(5R)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4]nonan-2 -yl]-3-methyl-butanoic acid (compound 4f 1) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-meth oxyethyl]- 5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa- 4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaene-8,14-dion e (intermediate H) instead of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4] nonan- 2-yl]-3-methyl-butanoic acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-21-(2,2,2- 25 913 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 31 (56.7 mg) was obtained as a yellow solid. MS calc’d 1063.5 (MH ⁺ ), measured 1063.5 (MH ⁺ ). 'HNMR (400 MHz, METHANOL-^) 6 = 8.67 (d, J = 7.6Hz, 1H), 8.48 (d, J = 2.8Hz, 1H), 7.65 (s, 1H), 7.57 (s, 1H), 7.34 (d, J =12.8Hz, 1H), 6.94 - 6.73 (m, 1H), 5.75 (t, J = 8.8Hz, 1H), 4.45 - 4.37 (m, 1H), 4.36 - 4.29 (m, 2H), 4.28 - 4.08 (m, 4H), 4.07 - 3.97 (m, 1H), 3.97 - 3.87 (m, 1H), 3.85 - 3.72 (m, 4H), 3.71 - 3.63 (m, 2H), 3.63 - 3.41 (m, 7H), 3.34 (s, 3H), 3.30 - 3.23 (m, 2H), 3.07 - 3.02 (m, 1H), 3.00 (s, 3H), 2.87 - 2.77 (m, 1H), 2.69 - 2.57 (m, 1H), 2.33 - 2.15 (m, 3H), 2.14 - 2.03 (m, 3H), 1.97 - 1.91 (m, 1H), 1.84 - 1.72 (m, 1H), 1.69 - 1.57 (m, 1H), 1.44 (d, J = 6.0Hz, 3H), 1.03 - 0.92 (m, 9H), 0.89 (d, J = 6.8Hz, 3H), 0.51 (s, 3H) ppm.

Example 32

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,1 3S)-21-ethyl-24-fluoro-(20M)-20-[2- [(lS)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-15- oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-2V-methyl-pyrroli dine- 3-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate H) instead of (7S, 13S)-7-ainino-25-fluoro-(20A4)-20-[2-[( lS)- l -mcthoxycthyl]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate E). Example 32 (36.7 mg) was obtained as a yellow solid. MS calc’d 1051.4 (MH ⁺ ), measured 1051.6 (MH ⁺ ). ^J H NMR (400 MHz, MeOD) 8 8.67 (d, J = 7.6 Hz, 1H), 8.49 (d, J = 2.8 Hz, 1H), 7.73 - 7.59 (m, 2H), 7.34 (d, J = 12.8 Hz, 1H), 6.96 - 6.70 (m, 1H), 5.82 - 5.61 (m, 1H), 5.00 - 4.90 (m, 2H), 4.83 - 4.77 (m, 1H), 4.49 - 4.38 (m, 1H), 4.37 - 4.05 (m, 5H), 4.00-3.89 (m, 1H), 3.89 - 3.47 (m, 10H), 3.47 - 3.39 (m, 2H), 3.36 (d, J = 4.8 Hz, 3H), 3.28 - 3.22 (m, 1H), 3.15 - 3.07 (m, 3H), 3.06 - 3.02 (m, 1H), 3.00 (s, 3H), 2.89 - 2.76 (m, 1H), 2.69 - 2.56 (m, 1H), 2.42 - 2.01 (m, 4H), 2.00 - 1.90 (m, 1H), 1.89 - 1.73 (m, 1H), 1.71 - 1.56 (m, 1H), 1.44 (d, J = 6.4 Hz, 3H), 1.25 - 0.96 (m, 6H), 0.96 - 0.92 (m, 3H), 0.92 - 0.82 (m, 3H), 0.61 - 0.39 (m, 3H) ppm.

Example 33

(3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,1 3S)-21-ethyl-24-fluoro-(20M)-20-[2-

[(lS)-l-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazi n-l-yl]-3-pyridyl]-17,17-dimethyl-

25 9 13 2226

8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17. 5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolid ine- 3-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (75,135)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15)-l-met hoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin- 1 -yl] -3 -pyridyl] - 17, 17-dimethyl- 15-oxa-4-thia-9,21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate L) instead of (7S,13S)-7-amino-25-fhioro-(20M)-20-[2-[(15)-l-methoxyethyl] -5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate E). Example 33 (45.2 mg) was obtained as a yellow solid. MS calc’d 1119.5 (MH ⁺ ), measured 1119.7 (MH ⁺ ). ^J H NMR (400 MHz, METHANOL-^) 6 = 8.73 - 8.65 (m, 1H), 8.38 (s, 1H), 7.94 - 7.86 (m, 1H), 7.72 - 7.62 (m, 1H), 7.37 (d, J = 12.6 Hz, 1H), 6.94 - 6.72 (m, 1H), 5.83 - 5.69 (m, 1H), 4.84 - 4.76 (m, 1H), 4.47 - 4.19 (m, 3H), 4.17 - 3.92 (m, 2H),

3.89 - 3.62 (m, 6H), 3.61 - 3.53 (m, 1H), 3.51 - 3.46 (m, 4H), 3.43 - 3.37 (m, 3H), 3.28 - 3.13 (m, 3H), 3.10 - 3.03 (m, 3H), 2.93 - 2.87 (m, 4H), 2.86 - 2.66 (m, 2H), 2.42 - 2.11 (m, 4H), 2.00 -

1.90 (m, 1H), 1.86 - 1.73 (m, 1H), 1.71 - 1.57 (m, 1H), 1.46 (d, J = 6.4 Hz, 3H), 1.36 - 1.27 (m, 1H), 1.14 - 0.78 (m, 13H), 0.69 - 0.54 (m, 3H) ppm.

Example 34 l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,13S)-21-e thyl-24-fluoro-(20M)-20-[2-[(lS)- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-15-oxa-4- thia-9, 21,27, 28-tetrazapentacyclo[17.5.2.1 ^2,5 .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25), 2, 5(28), 19, 22(26), 23- hexaen-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-A-methyl-pi peridine-4-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using 1- tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-21-ethyl-24- fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- 25 9 13 22226 dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ’ .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (S)-l-Boc- pyrrolidine-3-carboxylic acid (compound ID) and (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E). Example 34 (12.9 mg) was obtained as a yellow solid. MS calc’d 1083.5 (MEE), measured 1083.6 (MH ⁺ ), ^J H NMR (400 MHz, Methanol-^) 6 = 8.65 (d, J = 7.6Hz, 1H), 8.48 (d, J = 2.4Hz, 1H), 7.64 (s, 1H), 7.54 (s, 1H), 7.34 (d, J =12.4Hz, 1H), 7.11 - 6.95 (m, 1H), 5.78 - 5.71 (m, 1H), 4.47 - 4.37 (m, 2H), 4.35 - 4.23 (m, 2H), 4.22 - 4.07 (m, 4H), 4.00 - 3.88 (m, 2H), 3.80 - 3.69 (m, 3H), 3.68 - 3.57 (m, 2H), 3.54 - 3.41 (m, 4H), 3.34 (s, 3H), 3.20 (t, J = 4.8Hz, 4H), 3.14 - 3.05 (m, 2H), 3.03 - 2.95 (m, 5H), 2.86 - 2.79 (m, 1H), 2.65 - 2.57 (m, 1H), 2.32 - 2.13 (m, 5H), 2.03 - 1.90 (m, 2H), 1.84 - 1.77 (m, 1H), 1.68 - 1.59 (m, 1H), 1.44 (d, J = 6.0Hz, 3H), 1.29 - 1.09 (m, 2H), 1.00 - 0.97 (m, 3H), 0.93 (s, 3H), 0.91 - 0.86 (m, 3H), 0.50 (d, J = 5.2Hz, 3H).

Example 35 and Example 38 (2S)-2-[(5/?)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7-d iazaspiro[4.4]nonan-2-yl]-2V- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethy l]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]-3-methyl-butanamide (Example 35)

(2S)-2-[(5S)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-2V- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethy l]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]-3-methyl-butanamide (Example 38) Example 35 was prepared in analogy to the preparation of Example 4 by using (75,135)-7- armno-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]- 5-[4-(2,2,2- trifluoroethyl)piperazin- 1 -yl] -3 -pyridyl] - 17, 17-dimethyl- 15-oxa-4-thia-9,21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate L) instead of (7S,13S)-7-amino-24-fhioro-(20M)-20-[2-[(15)-l-methoxyethyl] -5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D). Example 35 (21.1 mg) was obtained as a yellow solid. MS calc’d 1131.5 (MH ⁺ ), measured 1131.6 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 8.71 (dd, J = 2.4,

7.6 Hz, 1H), 8.38 (t, J = 2.4 Hz, 1H), 7.87 - 7.81 (m, 1H), 7.76 - 7.62 (m, 1H), 7.37 (dd, J = 2.8,

12.6 Hz, 1H), 6.92 - 6.69 (m, 1H), 5.84 - 5.71 (m, 1H), 4.45 - 4.12 (m, 5H), 4.10 - 4.00 (m, 1H),

3.96 - 3.69 (m, 5H), 3.60 - 3.54 (m, 2H), 3.51 - 3.42 (m, 6H), 3.41 - 3.37 (m, 3H), 3.26 - 3.12 (m, 3H), 3.09 - 2.96 (m, 1H), 2.93 - 2.87 (m, 4H), 2.85 - 2.66 (m, 2H), 2.34 - 2.15 (m, 3H), 2.12 - 1.93 (m, 4H), 1.86 - 1.73 (m, 1H), 1.71 - 1.57 (m, 1H), 1.45 (d, J = 6.4 Hz, 3H), 1.04 - 0.95 (m, 9H), 0.89 (d, J = 6.8 Hz, 3H), 0.58 (d, J = 12.8 Hz, 3H).

Example 38 was prepared in analogy to the preparation of Example 4 by using (25)-2- [(5R)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4]nonan-2 -yl]-3-methyl-butanoic acid (compound 4f 1) and (7S,13S)-7-amino-21-ethyl-24-fhioro-(20Af)-20-[2-[(15)-l-met hoxyethyl]- 5-[4-(2,2,2-trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17- dimethyl-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate L) instead of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4] nonan-

2-yl]-3-methyl-butanoic acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-21-(2,2,2-

25 913 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 38 (26.1 mg) was obtained as a yellow solid. MS calc’d 1131.5 (MH ⁺ ), measured 1131.7 (MH ⁺ ). ' HNMR (400 MHz, METHANOL-^) 6 = 8.88 - 8.81 (m, 1H), 8.71 (d, J = 7.6Hz, 1H), 8.38 (d, J = 2.0Hz, 1H),

7.96 - 7.87 (m, 1H), 7.67 - 7.64 (m, 1H), 7.37 (d, J = 12.4Hz, 1H), 6.91 - 6.75 (m, 1H), 5.84 -

5.79 (m, 1H), 4.46 - 4.36 (m, 2H), 4.33 - 4.25 (m, 2H), 4.18 - 4.12 (m, 1H), 4.08 - 3.84 (m, 2H),

3.81 - 3.43 (m, 4H), 3.69 - 3.61 (m, 2H), 3.52 - 3.48 (m, 5H), 3.40 (s, 3H), 3.23 - 3.15 (m, 3H),

3.03 - 2.97 (m, 1H), 2.91 - 2.88 (m, 4H), 2.83 - 2.73 (m, 2H), 2.27 - 2.15 (m, 3H), 2.11 - 2.04 (m,

3H), 2.00 - 1.91 (m, 2H), 1.85 - 1.76 (m, 1H), 1.70 - 1.61 (m, 1H), 1.46 (d, J = 6.4Hz, 3H), 1.04 -

0.99 (m, 5H), 0.96 (s, 3H), 0.89 (d, J = 6.8Hz, 3H), 0.61(s, 3H) ppm.

Example 36

(2.S)-2-|9-|(2/?)-2-chloro-2-fluoro-acetyl|-l -oxo-2, 9-diazaspiro|5.5]undecan-2-yl|-V- [(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyeth yl]-5-(4-methylpiperazin-l-yl)-

3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,2 7,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (7S,13S)-7-amino-24-fhioro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D). Example 36 (27.5 mg) was obtained as a yellow solid. MS calc’d 1091.5 (MH ⁺ ), measured 1091.5 (MH ⁺ ). ^J H NMR (400MHz, Methanol-^) d = 8.67 (d, J = 7.6 Hz, 1H), 8.48 (d, 7 = 2.8 Hz, 1H), 7.67 (d, J = 2.4 Hz, 1H), 7.61 - 7.51 (m, 1H), 7.34 (d, J = 12.8 Hz, 1H), 7.15 - 6.85 (m, 1H), 5.80 - 5.68 (m, 1H), 4.86 - 4.76 (m, 1H), 4.47 - 4.36 (m, 1H), 4.35 - 4.05 (m, 6H), 3.97 - 3.86 (m, 1H), 3.83 - 3.67 (m, 3H), 3.66 - 3.51 (m, 2H), 3.50 - 3.40 (m, 4H), 3.34 (s, 3H), 3.30 - 3.09 (m, 4H), 3.07 - 3.01 (m, 1H), 3.00 (s, 3H), 2.88 - 2.76 (m, 1H), 2.68 - 2.56 (m, 1H), 2.33 - 2.08 (m, 4H), 1.97 - 1.75 (m, 6H), 1.68 - 1.57 (m, 3H), 1.44 (d, 7 = 6.0 Hz, 3H),1.37 - 1.24 (m, 2H), 1.01 - 0.90 (m, 9H), 0.87 (d, 7 = 6.4 Hz, 3H), 0.51 (s, 3H) ppm.

Example 37

(2S)-2-[2-[(2/f)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-2V-[(7S,13S)-

21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-[ 4-(2,2,2-trifluoroethyl)piperazin- l-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,2 1,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octan -6-yl)-3-methyl-butanoic acid (compound 25H) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]- 5-[4-(2,2,2-trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17- dimethyl-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate L) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 37 (32.1 mg) was obtained as a yellow solid. MS calc’d 1117.5 (MH ⁺ ), measured 1117.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.69 (d, J = 7.6 Hz, 1H), 8.38 (d, J = 2.4 Hz, 1H), 7.74 (s, 1H), 7.66 (dd, J = 4.0 Hz, 24.4 Hz, 1H), 7.35 (d, J = 12.8 Hz, 1H), 6.68 (d, J = 57.6 Hz, 1H), 5.78 (s, 1H), 4.57 (t, J = 5.6 Hz, 1H), 4.45 - 4.22 (m, 6H), 4.19 - 4.11 (m, 1H), 4.11 - 4.01 (m, 2H), 3.82 - 3.68 (m, 2H), 3.45 (s, 6H), 3.37 (d, J = 6.8 Hz, 4H), 3.22 - 3.12 (m, 2H), 3.07 - 2.97 (m, 1H), 2.88 (s, 4H), 2.76 - 2.66 (m, 2H), 2.38 (s, 2H), 2.26 - 2.13 (m, 2H), 1.99 - 1.92 (m, 1H), 1.86 - 1.74 (m, 1H), 1.69 - 1.61 (m, 1H), 1.45 (d, J = 6.4 Hz, 3H), 1.34 - 1.23 (m, 1H), 1.05 - 0.98 (m, 6H), 0.95 (s, 3H), 0.87 (dd, J = 2.4 Hz, 6.4 Hz, 3H), 0.57 (s, 3H) ppm.

Example 39 (2S)-2-[8-[(2/f)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diazasp iro[4.5]decan-2-yl]-A^-[(7S,13S)- 21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-[4-( 2,2,2-trifluoroethyl)piperazin- l-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,2 1,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(8-tert-butoxycarbonyl-l-oxo-2,8-diazaspiro[4.5]decan -2-yl)-3-methyl-butanoic acid (compound 15F) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]- 5-[4-(2,2,2-trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17- dimethyl-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate L) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 39 (22.9 mg) was obtained as a yellow solid. MS calc’d 1145.5 (MH ⁺ ), measured 1145.5 (MH ⁺ ). ^J H NMR (400 MHz, METHANOL-^) 6 = 8.85 - 8.77(m, 1H), 8.69 (d, J = 7.6Hz, 1H), 8.39 (d, J = 2.8Hz, 1H), 7.72 - 7.64 (m, 2H), 7.37 - 7.33(m, 1H), 7.09 - 6.93 (m, 1H), 5.83 - 5.74 (m, 1H), 4.46 - 4.37 (m, 1H), 4.34 - 4.24 (m, 4H), 4.18 - 4.14 (m, 1H), 4.11 - 4.04 (m, 1H), 3.96 - 3.89 (m, 1H), 3.80 - 3.71 (m, 2H), 3.64 - 3.50 (m, 2H), 3.45 - 3.41 (m, 5H), 3.36 (s, 3H), 3.20 - 3.13 (m, 4H), 3.04 - 2.98 (m, 1H), 2.90 - 2.86 (m, 4H), 2.83 - 2.78 (m, 1H), 2.73 - 2.68 (m, 1H), 2.25 - 2.16 (m, 2H), 2.06 - 2.01 (m, 2H), 1.97 - 1.76 (m, 5H), 1.68 - 1.53 (m, 3H), 1.44 (d, J = 6.4Hz, 3H), 1.02 - 0.98 (m, 5H), 0.95 (s, 3H), 0.89 - 0.86 (m, 3H), 0.57 (s, 3H) ppm.

Example 40 (2/f)-2-[[l-[(2/f)-2-chloro-2-fluoro-acetyl]azetidin-3-yl]ox ymethyl]-A^-[(7S,13S)-21-ethyl-24- fluoro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperazi n-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 13 by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (7S,13S)-7-amino-24-fhioro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D). Example 40 (26.2 mg) was obtained as a yellow solid. MS calc’d 1010.5 (MH ⁺ ), measured 1010.4 (MH ⁺ ). ^J H NMR (400 MHz, METHANOL-^) 6 = 8.66 (t, J = 7.8 Hz, 1H), 8.50 (d, J = 2.4 Hz, 1H), 7.72 (br s, 1H), 7.63 (t, J = 2.9 Hz, 1H), 7.34 (dd, J = 2.9, 12.6 Hz, 1H), 6.69 - 6.44 (m, 1H), 5.83 (br t, J = 7.5 Hz, 1H), 5.01 - 4.92 (m, 1H), 4.63 - 3.86 (m, 11H), 3.84 - 3.65 (m, 3H), 3.64 - 3.53 (m, 3H), 3.45 (br d, J = 15.2 Hz, 3H), 3.36 (br d, J = 3.4 Hz, 3H), 3.31 - 3.22 (m, 3H), 3.10 - 2.94 (m, 4H), 2.81 (br t, J = 12.4 Hz, 1H), 2.65 (br t, J = 13.4 Hz, 1H), 2.52 - 2.37 (m, 1H), 2.19 (br d, J = 12.2 Hz, 1H), 1.96 (br d, J = 12.8 Hz, 1H), 1.90 - 1.71 (m, 2H), 1.70 - 1.54 (m, 1H), 1.45 (d, J = 6.2 Hz, 3H), 1.17 - 0.77 (m, 12H), 0.54 (br d, J = 9.3 Hz, 3H) ppm.

Example 41

(2S)-2-[[l-[(2/f)-2-chloro-2-fluoro-acetyl]azetidin-3-yl] oxymethyl]-2V-[(7S,13S)-21-ethyl-24- fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 13 by using tert-butyl 3-[(25)-2-[[(75,135)-21-ethyl-24-fluoro-(20Af)-20-[2-[(15)-l -methoxyethyl]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-1 5-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carboxylate, compound 40B) instead of tert-butyl 3- [(27?)-2-[[(75,13S)-24-fhioro-(20M)-20-[2-[(15)-l-methoxyeth yl]-5-(4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]-octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7- yl]carbamoyl]-3-methyl-butoxy]azetidine-l-carboxylate (compound 131). Example 41 (32.9 mg) was obtained as a yellow solid. MS calc’d 1010.5 (MH ⁺ ), measured 1010.4 (MH ⁺ ). ^J H NMR (400 MHz, METHANOL-^) 6 = 8.68 (d, J = 7.4 Hz, 1H), 8.51 (d, J = 2.8 Hz, 1H), 7.80 (d, J = 2.8 Hz, 1H), 7.65 (d, J = 2.4 Hz, 1H), 7.35 (d, J = 12.6 Hz, 1H), 6.75 - 6.53 (m, 1H), 5.86 (br d, J = 7.1 Hz, 1H), 5.03 - 4.92 (m, 1H), 4.54 - 4.01 (m, 11H), 4.01 - 3.83 (m, 2H), 3.81 - 3.66 (m, 3H), 3.66 - 3.50 (m, 4H), 3.50 - 3.38 (m, 3H), 3.36 (s, 3H), 3.02 (br s, 1H), 2.99 (s, 3H), 2.86 - 2.74 (m, 1H), 2.74 - 2.62 (m, 1H), 2.48 - 2.34 (m, 1H), 2.17 (br d, J = 12.1 Hz, 1H), 2.00 - 1.85 (m, 2H), 1.83 - 1.70 (m, 1H), 1.68 - 1.55 (m, 1H), 1.45 (d, J = 6.3 Hz, 3H), 1.09 - 0.85 (m, 12H), 0.57 (s, 3H) ppm.

Compound 40B was prepared in analogy to the preparation of compound 13J by using (7S,13S)-7-amino-21-ethyl-24-fhioro-(20Af)-20-[2-[(15)-l-met hoxyethyl]-5-(4-methylpiperazin- l-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate H) instead of (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')-l-methoxyethy l]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia-

9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D).

Example 42

(2S)-2-[9-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,9-diaz aspiro[5.5]undecan-2-yl]-2V- [(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-[4- (2,2,2-trifluoroethyl)piperazin- l-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluo roethyl)-15-oxa-4-thia-

9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]-3-methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (75,135)-7-amino-24-fluoro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-21- (2,2,2-trifluoroethyl)-15-oxa-4-thia-

9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate J) instead of (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 42 (7.8 mg) was obtained as a yellow solid. MS calc’d 1213.5 (MEE), measured 1213.4 (MH ⁺ ). ^J H NMR (400MHz, Methanol-^) 3 = 8.70 (d, J = 7.6 Hz, 1H), 8.42 (d, J = 2.8 Hz, 1H), 7.74 - 7.67 (m, 1H), 7.52 - 7.41 (m, 2H), 7.13 - 6.87 (m, 1H), 5.75 - 5.65 (m, 1H), 5.20 - 5.11 (m, 1H), 4.81 - 4.78 (m, 1H), 4.48 - 4.34 (m, 1H), 4.25 - 4.05 (m, 3H), 3.98 - 3.65 (m, 3H), 3.52 - 3.33 (m, 11H), 3.30 - 3.09 (m, 6H), 2.91 - 2.78 (m, 5H), 2.68 - 2.57 (m, 1H), 2.32 - 2.06 (m, 4H), 2.00 - 1.77 (m, 6H), 1.70 - 1.57 (m, 3H), 1.44 (d, J = 6.0 Hz, 3H), 1.03 - 0.92 (m, 6H), 0.90 - 0.84 (m, 3H), 0.47 (s, 3H) ppm.

Example 43 and Example 45

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-N- [(7S,13S)-24-fluoro-20-[2-[(lS)-l-methoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 43) (Example 43)

(2S)-2-[(5S)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-2V- [(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-[4- (2,2,2-trifluoroethyl)piperazin- l-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluo roethyl)-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]-3-methyl-butanamide (Example 45)

(Example 45)

Example 43 prepared in analogy to the preparation of Example 4 by using (75,135)-7- amino-24-fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-[4-(2,2 ,2-trifluoroethyl)piperazin-l-yl]-

3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-ox a-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate J) instead of (7S,13S)-7-amino-24-fhioro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D). Example 43 (43.9 mg) was obtained as a yellow solid. MS calc’d 1185.4 (MH ⁺ ), measured 1185.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 8.82 - 8.71 (m, 1H), 8.44 (br s, 1H), 7.84 - 7.66 (m, 1H), 7.56 - 7.45 (m, 2H), 6.96 - 6.71 (m, 1H), 5.77 - 5.65 (m, 1H), 5.26 - 5.16 (m, 1H), 4.49 - 4.33 (m, 2H), 4.31 - 4.19 (m, 2H), 4.07 - 3.69 (m, 5H), 3.66 - 3.55 (m, 2H), 3.54 - 3.34 (m, 10H), 3.25 - 3.12 (m, 3H), 2.96 - 2.78 (m, 5H), 2.72 - 2.57 (m, 1H), 2.40 - 2.17 (m, 3H), 2.16 - 1.92 (m, 4H), 1.91 - 1.76 (m, 1H), 1.75 - 1.57 (m, 1H), 1.53 - 1.41 (m, 3H), 1.38 - 1.14 (m, 1H), 1.06 - 0.97 (m, 6H), 0.94 - 0.86 (m, 3H), 0.59 - 0.42 (m, 3H) ppm.

Example 45 was prepared in analogy to the preparation of Example 4 by using (25)-2- [(5R)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4]nonan-2 -yl]-3-methyl-butanoic acid (compound 4f 1) and (7S, 13S)-7-ainino-24-fluoro-(20A7)-20-[2-[( lS)- l -mcthoxycthyl]-5-[4- (2,2,2-trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimet hyl-21-(2,2,2-trifluoroethyl)-15-oxa-

4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaene- 8, 14-dione (Intermediate J) instead of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7- diazaspiro[4.4]nonan-2-yl]-3-methyl-butanoic acid (compound 4f 2) and (7S,13S)-7-amino-24- fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyc lo[ 17.5.2^ ² ’^ I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28) J9, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 45 (24.2 mg) was obtained as a white solid. MS calc’d 1185.4 (MH ⁺ ), measured 1185.5 (MH ⁺ ). ' HNMR (400 MHz, METHANOL-^) 6 = 8.70 (d, J = 7.6 Hz, 1H), 8.42 (d, 7 = 2.8 Hz, 1H), 7.71 (t, 7 = 2.4 Hz, 1H), 7.57 (s, 1H), 7.48 (d, 7 = 12.8 Hz, 1H), 6.84 (dd, 7 = 10.8 Hz, 48.8 Hz, 1H), 5.72 (s, 1H), 5.24 - 5.21 (m, 2H), 4.48 - 4.18 (m, 3H), 3.82-3.68 (m, 4H), 3.68-3.55 (m, 2H), 3.55 - 3.44 (m, 3H), 3.42 (d, 7 = 3.6 Hz, 4H), 3.37 (s, 3H), 3.30 - 3.08 (m, 5H), 2.89 (t, 7 = 4.8 Hz, 4H), 2.86 - 2.77 (m, 1H), 2.64 (d, 7 = 14.4 Hz, 1H), 2.32 - 2.15 (m, 3H), 2.14 - 2.01 (m, 3H), 1.98 - 1.90 (m, 1H), 1.88 - 1.74 (m, 1H), 1.69 - 1.57 (m, 1H), 1.45 (d, 7 = 6.4 Hz, 3H), 1.01 (d, 7 = 6.4 Hz, 3H), 0.99 (s, 3H), 0.90 (d, 7 = 6.4 Hz, 3H), 0.49 (s, 3H) ppm.

Example 44 l-[(2/?)-2-chloro-2-fluoro-acetyl]-4-fluoro-^-[(lS)-l-[[(7S, 13S)-24-fluoro-(20M)-20-[2-[(lS)- l-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-l-yl]-3 -pyridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamoyl]-2-methyl-propyl]-2V-methyl-piperidine-4-carbox amide

The title compound was prepared in analogy to the preparation of Example 1 by using 1- tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-24-fluoro- (20A7)-20-[2-[(15)-l-methoxyethyl]-5-[4-(2,2,2-trifluoroethy l)piperazin-l-yl]-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate J) instead of (5)- l-Boc-pyrrolidine-3 -carboxylic acid (compound ID) and (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2-[(15')-l-methoxyethy l]-5-(4-methylpiperazin-l-yl)-3- pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-t hia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E). Example 44 (18.4 mg) was obtained as an off-white solid. MS calc’d 1205.4 (MH ⁺ ), measured 1205.6 (MH ⁺ ), ^J H NMR (400 MHz, Methanol-^) 6 = 8.69 (d, J = 7.2Hz, 1H), 8.42 (d, J = 2.8Hz, 1H), 7.69 (s, 1H), 7.58 - 7.51 (m, 1H), 7.48 (d, J =12.4Hz, 1H), 7.11 - 6.97 (m, 1H), 5.74 - 5.68 (m, 1H), 5.24 - 5.10 (m, 2H), 4.75 (d, J = 11.2Hz, 1H), 4.46 - 4.34 (m, 2H), 4.27 - 4.16 (m, 2H), 4.00 - 3.88 (m, 1H), 3.81 - 3.69 (m, 2H), 3.57 - 3.49 (m, 1H), 3.47 - 3.38(m, 5H), 3.37 (s, 3H), 3.22 - 3.17 (m, 4H), 3.16 - 3.12 (m, 2H), 3.10 - 3.01 (m, 1H), 2.88 (t, J = 4.4Hz, 5H), 2.84 - 2.77 (m, 1H), 2.68 - 2.59 (m, 1H), 2.35 - 2.15 (m, 6H), 1.99 - 1.93 (m, 1H). 1.88 - 1.77 (m, 1H), 1.69 - 1.59 (m, 1H), 1.45 (d, J = 6.0Hz, 3H), 1.00 - 0.95 (m, 6H), 0.89 (d, J = 6.4Hz, 3H), 0.48 (d, J = 5.6Hz, 3H).

Example 46

(2S)-2-[2-[(2/f)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-A^-[(7S,13S)- 24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-[4-(2,2,2-tri fluoroethyl)piperazin-l-yl]-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octan -6-yl)-3-methyl-butanoic acid (compound 25H) and (7S, 13S)-7-ainino-24-fluoro-(20A4)-20-[2-[( lS)- l -mcthoxycthyl]-5-[4- (2,2,2-trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimet hyl-21-(2,2,2-trifluoroethyl)-15-oxa- 4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaene- 8, 14-dione (Intermediate J) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9- diazaspiro[5.5]undecan-2-yl)-3-methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24- fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 46 (17.0 mg) was obtained as an off-white solid. MS calc’d 1171.4 (MH ⁺ ), measured 1171.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.72 (d, J = 7.6 Hz, 1H), 8.42 (d, J = 2.4 Hz, 1H), 7.74 (d, J = 2.4 Hz, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.50 (d, J = 12.4Hz, 1H), 6.69 (dd, J = 1.6 Hz, 48.8 Hz, 1H), 5.74 - 5.69 (m, 1H), 4.44 - 4.21 (m, 5H), 3.84 - 3.67 (m, 2H), 3.59-3.52 (m, 1H), 3.46 (s, 6H), 3.40 (d, J = 1.2 Hz, 3H), 3.24-3.14 (m, 3H), 3.14 - 3.08 (m, 1H), 2.90 (t, J = 5.2 Hz, 4H), 2.85 - 2.77 (m, 1H), 2.71 - 2.61 (m, 1H), 2.44 - 2.36 (m, 2H), 2.27 - 2.14 (m, 2H), 2.01 - 1.90 (m, 1H), 1.90 - 1.71 (m, 1H), 1.70 - 1.55 (m, 1H), 1.47 (d, J = 6.4 Hz, 3H), 1.25 (s, 2H), 1.20 (s, 3H), 1.02 - 0.96 (m, 6H), 0.88 (dd, J = 3.2 Hz, 6.4 Hz, 3H), 0.51 (s, 3H) ppm.

Example 47

(2S)-2-[8-[(2/f)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diaz aspiro[4.5]decan-2-yl]-A^-[(7S,13S)- 24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-[4-(2,2,2-tri fluoroethyl)piperazin-l-yl]-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(8-tert-butoxycarbonyl-l-oxo-2,8-diazaspiro[4.5]decan -2-yl)-3-methyl-butanoic acid (compound 15F) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')-l-methoxyethy l]-5-[4- (2,2,2-trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimet hyl-21-(2,2,2-trifluoroethyl)-15-oxa- 4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaene- 8, 14-dione (Intermediate J) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9- diazaspiro[5.5]undecan-2-yl)-3-methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24- fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiperaz in-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 47 (21.1 mg) was obtained as a white solid. MS calc’d 1199.5 (MEE), measured 1199.4 (MH ⁺ ), ^J H NMR (400 MHz, Methanol-^) 6 = 8.86 - 8.79 (m, 1H), 8.69 (d, J = 7.2 Hz, 1 H), 8.44 - 8.40 ( m, 1H), 7.69 (s, 1H), 7.51 - 7.43 (m, 2H), 7.12 - 6.90 (m, 1H), 5.69 (s, 1H), 5.22 - 5.11 (m, 1H), 4.44 - 4.37 (m, 1H), 4.34 - 4.19 (m, 4H), 3.99 - 3.87 (m, 1H), 3.81 - 3.68 (m, 2H), 3.62 - 3.58 (m, 1H), 3.49 - 3.42 (m, 2H), 3.41 - 3.37 ( m, 4H), 3.35 (s, 3H), 3.28 - 3.23 (m, 1H), 3.20 - 3.09 (m, 4H), 2.90 - 2.86 (m, 4H), 2.84 - 2.78 (m, 1H), 2.62 (d, J = 14.4 Hz, 1H), 2.26 - 2.19 (m, 2H), 2.10 - 2.02 (m, 2H), 2.00 - 1.90 (m, 2H), 1.87 - 1.76 (m, 2H), 1.70 - 1.53 (m, 3H), 1.44 (d, 7 = 6.0 Hz, 3H), 1.18 (t, 7 = 7.2 Hz, 1H), 1.03 - 0.96 (m, 6H), 0.88 ( d, 7 = 5.6 Hz, 3H), 0.47 ( s, 3H) ppm.

Example 48

(2.S)-2-|9-|(2/?)-2-chloro-2-fhioro-acetyl|-l -oxo-2, 9-diazaspiro|5.5]undecan-2-yl|-V- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(lS)-l-methoxyethy l]-5-[4-(2,2,2- trifluoroethyl)piperazin-l-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]-3-methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (75,135)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15)-l-met hoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin- 1 -yl] -3 -pyridyl] - 17, 17-dimethyl- 15-oxa-4-thia-9,21 ,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate L) instead of (7S,13S)-7-amino-24-fhioro-(20Af)-20-[2-[(15)-l-methoxyethyl ]-5- (4-methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2 -trifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate D). Example 48 (10 mg) was obtained as a yellow solid. MS calc’d 1159.5 (MH ⁺ ), measured 1159.5 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.69 (d, J = 7.6 Hz, 1H), 8.40 (d, J = 2.8 Hz, 1H), 7.68 - 7.64 (m, 1H), 7.63 - 7.57 (m, 1H), 7.35 (d, J = 12.8 Hz, 1H), 7.11 - 6.87 (m, 1H), 5.82 - 5.71 (m, 1H), 4.82 - 4.75 (m, 2H), 4.45 - 4.38 (m, 1H), 4.36 - 4.29 (m, 1H), 4.27 - 4.03 (m, 4H), 3.97 - 3.81 (m, 1H), 3.79 - 3.70 (m, 2H), 3.47 - 3.42 (m, 6H), 3.36 (s, 3H), 3.30 - 3.12 (m, 5H), 3.08 - 2.96 (m, 1H), 2.92 - 2.79 (m, 5H), 2.73 - 2.64 (m, 1H), 2.32 - 2.06 (m, 4H), 2.00 - 1.90 (m, 1H), 1.90 - 1.72 (m, 5H), 1.68 - 1.55 (m, 3H), 1.44 (d, J = 6.4 Hz, 3H), 1.05 - 0.94 (m, 9H), 0.89 - 0.82 (m, 3H), 0.55 (s, 3H) ppm.

Example 49 l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,13S)-21-e thyl-24-fluoro-(20M)-20-[2-[(lS)- l-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-l-yl]-3 -pyridyl]-17,17-dimethyl-8,14- 25 9 13 2226 dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ’ .1 ’ .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-4-fhioro-2V-methy l- piperidine-4-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using 1- tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-21-ethyl-24- fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-[4-(2,2,2-triflu oroethyl)piperazin-l-yl]-3-pyridyl]- 25 9 13 22226 17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17 .5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate L) instead of (S)-l-Boc- pyrrolidine-3-carboxylic acid (compound ID) and (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-21-(2,2,2- 25 9 13 2226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate E). Example 49 (25.1 mg) was obtained as a yellow solid. MS calc’d 1151.5 (MEE), measured 1151.5 (MH ⁺ ), ^J H NMR (400 MHz, Methanol-^) 6 = 8.70 (d, J = 7.6 Hz, 1H), 8.39 (d, J = 2.8 Hz, 1H), 7.85 - 7.78 (m, 1H), 7.67 - 7.60 (m, 1H), 7.36 (d, J = 12.4 Hz, 1H), 7.11 - 6.94 (d, J = 49.2 Hz, 1H), 5.89 - 5.69 (m, 1H), 4.77 - 4.67 (m, 1H), 4.46 - 4.32 (m, 3H), 4.30 - 4.21 (m, 1H), 4.18 - 4.12 (m, 1H), 4.10 - 4.00 (m, 1H), 3.96 - 3.86 (m, 1H), 3.81 - 3.71 (m, 2H), 3.51 - 3.43 (m, 5H), 3.43 - 3.34 (m, 4H), 3.28 - 3.13 (m, 7H), 3.03 - 2.98 (m, 1H), 2.92 - 2.87 (m, 4H), 2.86 - 2.78 (m, 1H), 2.76 - 2.66 (m, 1H), 2.36 - 2.08 (m, 6H), 1.99 - 1.90 (m, 1H), 1.85 - 1.71 (m, 1H), 1.70 - 1.58 (m, 1H), 1.45 (d, J = 6.4 Hz, 3H), 1.04 - 0.93 (m, 9H), 0.91 - 0.80 (m, 3H), 0.65 - 1.53 m, 3H) ppm.

Example 50

(2S)-2-[8-[(2/f)-2-chloro-2-fluoro-acetyl]-l-oxo-2,8-diaz aspiro[4.5]decan-2-yl]-A^-[(7S,13S)-

21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-m orpholino-3-pyridyl]-17,17- dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(8-tert-butoxycarbonyl-l-oxo-2,8-diazaspiro[4.5]decan -2-yl)-3-methyl-butanoic acid (compound 15F) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]- 5-morpho lino-3-pyridyl]- 17, 17-dimethyl-15-oxa-4-thia-9, 21,27, 28-tetrazapentacyc loin.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate M) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2-yl)-3-methyl- butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 50 (51.8 mg) was obtained as a yellow solid. MS calc’d 1064.5 (MH ⁺ ), measured 1064.3 (MH ⁺ ), ^J H NMR (400 MHz, CHLOROFORM-7) 8 = 8.99 (br s, 1H), 8.66 - 8.64 (d, J = 7.5 Hz, 1H), 7.60 - 7.58 (dd, J = 1.8, 8.4 Hz, 1H), 7.45 - 7.41 (dd, 7 = 2.3, 16.1 Hz, 1H), 7.32 - 7.30 (br d, J = 8.9 Hz, 0.5H), 7.19 - 7.18 (br d, 7 = 8.8 Hz, 0.5H), 7.14 - 7.11 (d, 7 = 12.3 Hz, 1H), 6.57 - 6.54 (m, 0.5H), 6.40 - 6.28 (m, 0.5H), 5.81 (td, 7 = 9.2, 13.9 Hz, 1H), 4.66 - 4.52 (m, 1H), 4.40 (q, 7 = 6.4 Hz, 1H), 4.31 - 4.22 (m, 1H), 4.19 - 4.03 (m, 5H), 3.92 (br t, 7 = 4.5 Hz, 4H), 3.83 (br d, 7 = 11.2 Hz, 1H), 3.69 (br d, 7 = 11.1 Hz, 1H), 3.62 - 3.54 (m, 8H), 3.45 - 3.36 (m, 9H), 3.21 - 3.07 (m, 2H), 2.78 - 2.65 (m, 1H), 2.50 - 2.30 (m, 2H), 2.22 (br d, 7 = 12.0 Hz, 1H), 2.00 - 1.96 (m, 2H), 1.92 - 1.76 (m, 1H), 1.61 (br d, 7 = 10.1 Hz, 2H), 1.55-1.54 (d, 7 = 6.4 Hz, 3H), 1.06 - 0.93 (m, 9H), 0.89- 0.87 (dd, 7 = 2.4, 6.4 Hz, 3H), 0.49 (s, 3H) ppm.

Example 51 and Example 53

(2S)-2-[(5S)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2,7 -diazaspiro[4.4]nonan-2-yl]-2V-

[(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxy ethyl]-5-morpholino-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 51)

(Example 51)

(2S)-2-[(5/?)-7-[(2/?)-2-chloro-2-fluoro-acetyl]-l-oxo-2, 7-diazaspiro[4.4]nonan-2-yl]-2V-

[(7S,13S)-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxy ethyl]-5-morpholino-3-pyridyl]-

17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide (Example 53)

(Example 53)

Example 51 was prepared in analogy to the preparation of Example 4 by using (25)-2- [(57?)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4]nonan- 2-yl]-3-methyl-butanoic acid (compound 4f 1) and (7SJ3S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-meth oxyethyl]-

5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21 ,27,28-tetrazapentacyclo- [17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate M) instead of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4] nonan-2-yl]-3-methyl- butanoic acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 25 9 13 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 51 (21.4 mg) was obtained as a yellow solid. MS calc’d 1050.4 (MH ⁺ ), measured 1050.4 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.70 (d, J = 7.6 Hz, 1H), 8.39 (d, J = 2.8 Hz, 1H), 7.89 (dd, J = 2.8, 9.6 Hz, 1H), 7.65 (s, 1H), 7.38 (d, J = 12.8 Hz, 1H), 8.90 - 6.74 (m, 1H), 5.79 (d, J = 9.2 Hz, 1H),

4.44 - 4.37 (m, 2H), 4.33 - 4.24 (m, 2H), 4.18 - 4.12 (m, 1H), 4.08 - 4.00 (m, 1H), 3.87 (t, J = 4.4 Hz, 4H), 3.81 - 3.70 (m, 4H), 3.68 - 3.63 (m, 1H), 3.58 - 3.54 (m, 1H), 3.51 - 3.47 (m, 1H),

3.45 - 3.41 (m, 5H), 3.40 (s, 3H), 3.29 - 3.23 (m, 1H), 3.02 (d, J = 14.4 Hz, 1H), 2.86 - 2.71 (m, 2H), 2.28 - 2.15 (m, 3H), 2.09 - 2.00 (m, 3H), 1.98 - 1.93 (m, 1H), 1.84 - 1.75 (m, 1H), 1.68 - 1.60 (m, 1H), 1.46 (d, J = 6.4 Hz, 3H), 1.34 - 1.28 (m, 1H), 1.04 - 0.99 (m, 6H), 0.97 (s, 3H), 0.88 (d, J = 6.4 Hz, 3H), 0.60 (s, 3H).

Example 53 prepared in analogy to the preparation of Example 4 by using (75,135)-7- amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-methoxyethyl] -5-morpholino-3-pyridyl]-17,17- 25 9 13 2226 dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo-[17.5.2. 1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate M) instead of (7S,13S)-7-amino- 24-fhioro-(20M)-20-[2-[(lS)-l-methoxyethyl]-5-(4-methylpiper azin-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 53 (28.8 mg) was obtained as a yellow solid. MS calc’d 1050.4 (MH ⁺ ), measured 1050.5 (MH ⁺ ). ¹ H NMR (400 MHz, Methanol-^) 6 = 8.88 - 8.87 (m, 1H), 8.72

- 8.68 (m, 1H), 8.38 (t, J = 2.4Hz, 1H), 7.80 (t, J = 2.8Hz, 1H), 7.74 - 7.64 (m, 1H), 7.39 - 7.34 (m, 1H), 6.91 - 6.69 (m, 1H), 5.81 - 5.73 (m, 1H), 4.45 - 4.36 (m, 2H), 4.34 - 4.30 (m, 1H), 4.29

- 4.23 (m, 1H), 4.21 - 4.13 (m, 1H), 4.11 - 4.02 (m, 1H), 3.93 - 3.85 (m, 5H), 3.81 - 3.72 (m, 4H), 3.63 - 3.51 (m, 3H), 3.49 - 3.43 (m, 2H), 3.42 - 3.38 (m, 7H), 3.09 - 2.99 (m 1H), 2.87 - 2.78 (m, 1H), 2.70 (t, J = 14.0 Hz, 1H), 2.33 - 2.25 (m, 1H), 2.24 - 2.16 (m, 2H), 2.12 - 2.06 (m, 2H), 2.02

- 1.91 (m, 2H), 1.85 - 1.76 (m, 1H), 1.70 - 1.61 (m, 1H), 1.46 (d, 7 = 6.4 Hz, 3H), 1.04 - 0.98 (m, 6H), 0.97 - 0.96 (m, 2H), 0.89 (d, J = 6.8 Hz, 3H), 0.61 - 0.55 (m, 3H).

Example 52 (2S)-2-[2-[(2/f)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazasp iro[3.4]octan-6-yl]-A^-[(7S,13S)-

21-ethyl-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-m orpholino-3-pyridyl]-17,17- dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octan -6-yl)-3-methyl-butanoic acid (compound 25H) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]- 5-morpho lino-3-pyridyl]- 17, 17-dimethyl-15-oxa-4-thia-9, 21,27, 28-tetrazapentacyc loin.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate M) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2-yl)-3-methyl- butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l- methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2-trifluoroethyl)- 25 913 22226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 52 (16.7 mg) was obtained as a yellow solid. MS calc’d 1036.4 (MH ⁺ ), measured 1036.4 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.69 (d, J = 7.2 Hz, 1H), 8.38 (d, J = 2.4 Hz, 1H), 7.85 - 7.82 (m, 1H), 7.69 - 7.63 (m, 1H), 7.37 (d, J = 12.4 Hz, 1H), 6.75 - 6.62 (m, 1H), 5.80 - 5.74 (m, 1H), 4.57 (t, J = 10.0 Hz, 1H), 4.41 - 4.34 (m, 3H), 4.32 - 4.26 (m, 2H), 4.19 - 4.13 (m, 1H), 4.10 - 4.03 (m, 2H), 3.87 (t, J = 4.8 Hz, 4H), 3.80 - 3.71 (m, 2H), 3.53 - 3.46 (m, 2H), 3.42 - 3.41 (m, 3H), 3.39 - 3.38 (m, 3H), 3.07 - 2.98 (m, 1H), 2.87 - 2.79 (m, 1H), 2.75 - 2.68 (m, 1H), 2.41 - 2.35 (m, 2H), 2.28 - 2.14 (m, 3H), 2.07 - 1.90 (m, 2H), 1.85 - 1.76 (m, 1H), 1.66 - 1.60 (m, 1H), 1.46 (d, J = 6.4 Hz, 3H), 1.33 - 1.29 (m, 2H), 1.03 - 0.98 (m, 6H), 0.98 - 0.94 (m, 3H), 0.89 - 0.85 (m, 3H), 0.58 (s, 3H). Example 54 l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,13S)-21-e thyl-24-fluoro-(20M)-20-[2-[(lS)- l-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14- dioxo-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-2V-methyl-p iperidine-4-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using 1- tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-21-ethyl-24- fluoro-(20A7)-20-[2-[(lS)-l-methoxyethyl]-5-morpholino-3-pyr idyl]-17,17-dimethyl-15-oxa-4- thia-9,21,27,28-tetrazapentacyclo-[17.5.2.1 ² ’ ⁵ .l ^{9 13} .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaene- 8, 14-dione (Intermediate M) instead of (S)-l-Boc-pyrrolidine-3-carboxylic acid (compound ID) and (7S,13S)-7-amino-25-fluoro-(20A7)-20-[2-[(15')-l-methoxyethy l]-5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (intermediate E). Example 54 (44.2 mg) was obtained as a yellow solid. MS calc’d 1070.5 (MH ⁺ ), measured 1070.3 (MH ⁺ ), ^J H NMR (400 MHz, Methanol-^) 6 = 8.70 (d, J = 7.2 Hz, 1H), 8.39 (d, J = 2.8 Hz, 1H), 7.90 - 7.83 (m, 1H), 7.64 (d, J = 2.4 Hz, 1H), 7.37 (d, J = 12.8 Hz, 1H), 7.12 - 6.95 (d, J = 49.2 Hz, 1H), 5.87 - 5.72 (m, 1H), 4.85 - 4.69 (m, 1H), 4.46 - 4.22 (m, 4H), 4.19 - 4.12 (m, 1H), 4.10 - 4.00 (m, 1H), 3.98 - 3.90 (m, 1H), 3.88 (t, J = 4.8 Hz, 4H), 3.81 - 3.71 (m, 2H), 3.55 - 3.47 (m, 1H), 3.45 - 3.38 (m, 8H), 3.30 - 3.20 (m, 2H), 3.20 - 3.13 (m, 3H), 3.06 - 2.98 (m, 1H), 2.89 - 2.67 (m, 2H), 2.35 - 2.07 (m, 6H), 1.99 - 1.90 (m, 1H), 1.86 - 1.73 (m, 1H), 1.72 - 1.57 (m, 1H), 1.46 (d, J = 6.4 Hz, 3H), 1.05 - 0.94 (m, 9H), 0.88 (d, J = 6.4 Hz, 3H), 0.66 - 0.55 (m, 3H).

Example 55 (3S)-l-[(2/?)-2-chloro-2-fluoro-acetyl]-^-[(lS)-l-[[(7S,13S) -21-ethyl-24-fluoro-(20M)-20-[2- [(lS)-l-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl -8,14-dioxo-15-oxa-4-thia- 9,21,27,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23- hexaen-7-yl (carbamoyl |-2-inethyl-propyl|- \-inethyl-pyrrolidine-3-carboxamide

The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20Af)-20-[2-[(15')-l-me thoxyethyl]-5-morpholino-3- pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapent acyclo-

[17.5.2.1 ² ’ ⁵ .1 ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate M) instead of (75,13S)-7-amino-25-fhioro-(20M)-20-[2-[(15)-l-methoxyethyl] -5-(4- methylpiperazin-l-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-tr ifluoroethyl)-15-oxa-4-thia- 9,21,27 ,28-tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25),2,5(28), 19, 22(26), 23-hexaene- 8, 14-dione (Intermediate E). Example 55 (24.7 mg) was obtained as a yellow solid. MS calc’d 1038.4 (MH ⁺ ), measured 1038.6 (MH ⁺ ). ^J H NMR (400 MHz, METHANOL-^) 6 = 8.73 - 8.68 (m, 1H), 8.41 - 8.36 (m, 1H), 7.91 - 7.82 (m, 1H), 7.70 - 7.67 (m, 1H), 7.37 (d, J =16.0 Hz, 1H), 6.92 - 6.72 (m, 1H), 5.81 - 5.70 (m, 1H), 4.81 - 4.71 (dd, J = 4.0 , 12.0 Hz, 1H), 4.47 - 4.33 (m,

2H), 4.32 - 4.22 (m, 1H), 4.21 - 4.11 (m, 1H), 4.09 - 4.00 (m, 1H), 3.97 - 3.85 (m, 5H), 3.82 -

3.72 (m, 4H), 3.69 - 3.54 (m, 2H), 3.46 - 3.37 (m, 8H), 3.28 - 3.21 (m, 1H), 3.15 -3.05 (m, 3H),

3.04 - 2.95 (m, 1H), 2.88 - 2.67 (m, 2H), 2.41 - 2.06 (m, 4H), 1.97 - 1.57 (m, 3H), 1.46 (d, J =

8.0 Hz, 3H), 1.08 - 0.93 (m, 9H), 0.86 (d, J = 4.0 Hz, 3H), 0.73 - 0.56 (m, 3H).

Example 57

(2.S)-2-|(5.S)-7-|(2/?)-2-chloro-2-fluoro-acetyl|-l -oxo-2, 7-diazaspiro|4.4]nonan-2-yl|- V- [(7S,13S)-24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-mor pholino-3-pyridyl]-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

Example 57 was prepared in analogy to the preparation of Example 4 by using (25)-2- [(5R)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4]nonan-2 -yl]-3-methyl-butanoic acid (compound 4f 1) and (7SJ 3S)-7-ainino-24-fluoro-(2(W)-20-[2-[( lS)- l -mcthoxycthyl]-5- morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethy l)-15-oxa-4-thia-9,21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .I ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (Intermediate I) instead of (2S)-2-[(5S)-7-tert-butoxycarbonyl-l-oxo-2,7-diazaspiro[4.4] nonan- 2-yl]-3-methyl-butanoic acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2- [(15)-l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]- 17,17-dimethyl-21-(2,2,2- 25 913 22226 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ’ ]octacosa- 1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 57 (26 mg) was obtained as a white solid. MS calc’d 1104.4 (MH ⁺ ), measured 1104.4 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.86 - 8.98 (d, J =7.6 Hz,l H) , 8.66 - 8.75 (d, 7=7.2 Hz,l H), 8.32 - 8.49 (d, 7=2.8 Hz,l H), 7.66 - 7.75 (t, 7=2.8 Hz, 1 H), 7.56 - 7.63 (m, 1 H), 7.40 - 7.54 (d, 7=12.4 Hz, 1 H), 6.73-6.94 (dd, 7i=l 1.2 Hz, 7 ₂ =49.2 Hz, 1 H), 5.60 - 5.81 (t, 7=8.4 Hz, 1 H), 5.11 - 5.31 (m, 1 H), 4.38 - 4.47 (d, 7=12.4 Hz, 1 H), 4.30 - 4.37 (dd, 7;=3.2 Hz, 7 ₂ =11.2 Hz, 1 H), 4.16 - 4.30 (m, 2 H), 3.82 - 4.00 (m, 4 H), 3.65 - 3.82 (m, 5 H), 3.55 - 3.64 (m, 2 H), 3.42 - 3.54 (m, 3 H), 3.33 - 3.40 (m, 7 H), 3.07 - 3.18 (m, 1 H), 2.76 - 2.90 (m, 1 H), 2.58 - 2.74 (m, 1 H), 2.16 - 2.33 (m, 3 H), 2.02 - 2.14 (m, 3 H), 1.89 - 2.00 (m, 2 H), 1.75 - 1.88 (m, 1 H), 1.56 - 1.71 (m, 1 H), 1.41 - 1.52 (d, 7=6.4 Hz, 3 H), 0.94 - 1.07 (m, 6 H), 0.86 - 0.92 (d, 7=6.4 Hz, 3 H), 0.43 - 0.54 (s, 3 H).

Example 58

(2S)-2-[2-[(2/f)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diaz aspiro[3.4]octan-6-yl]-A^-[(7S,13S)-

24-fluoro-(20Af)-20-[2-[(lS)-l-methoxyethyl]-5-morpholino -3-pyridyl]-17,17-dimethyl-8,14- dioxo-21-(2, 2, 2-trifluoroethyl)-15-oxa-4-thia-9, 21,27,28- tetrazapentacyclo[17.5.2.1 ² ’ ⁵ .l ⁹ ’ ¹³ .0 ²² ’ ²⁶ ]octacosa-l(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- methyl-butanamide

The title compound was prepared in analogy to the preparation of Example 6 by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octan -6-yl)-3-methyl-butanoic acid (compound 25H) and (7S,13S)-7-amino-24-fluoro-(20A7)-20-[2-[(15')-l-methoxyethy l]-5- morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethy l)-15-oxa-4-thia-9,21,27,28- 25 9 13 22226 tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-l(25),2,5(28), 19, 22(26), 23-hexaene-8, 14-dione

(Intermediate I) instead of (2S)-2-(9-tert-butoxycarbonyl-l-oxo-2,9-diazaspiro[5.5]undec an-2- yl)-3-methyl-butanoic acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20Af)-20-[2-[(15')- l-methoxyethyl]-5-(4-methylpiperazin-l-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)- 25 9 13 2226 15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ’ ]octacosa-

1(25), 2, 5(28), 19, 22(26), 23-hexaene-8, 14-dione (intermediate D). Example 58 (24.0 mg) was obtained as a white solid. MS calc’d 1090.4 (MH ⁺ ), measured 1090.4 (MH ⁺ ). ^J H NMR (400 MHz, Methanol-^) 6 = 8.76 - 8.91 (d, J = 8.4 Hz, 1 H), 8.56 - 8.64 (d, J = 7.6 Hz, 1 H), 8.28 - 8.36 (d, J = 2.4 Hz, 1 H), 7.57 - 7.67 (dd, J = 2.4 Hz, 17.6 Hz, 1 H), 7.44 - 7.52 (m, 1 H), 7.35 - 7.42 (d, J = 12.8 Hz, 1 H), 6.51 - 6.68 (dd, 7 = 3.2 Hz, 52.4 Hz, 1 H), 5.55 - 5.70 (t, J = 8.4 Hz,l H), 4.99 - 5.21 (m, 1H), 4.43 - 4.56 (m, 1 H), 4.08 - 4.38 (m, 6 H), 3.91 - 4.04 (m, 1 H), 3.73 - 3.84 (t, J = 4.8 Hz, 4 H), 3.66 - 3.72 (m, 1 H), 3.56 - 3.64 (m, 1 H), 3.42 - 3.50 (m, 1 H), 3.32 - 3.41 (m, 2 H), 3.23 - 3.30 (m, 8 H), 2.97 - 3.07 (m, 1 H), 2.65 - 2.78 (m, 1 H), 2.49 - 2.58 (m, 1 H), 2.24 - 2.35 (m, 2 H), 2.06 - 2.19 (m, 2 H), 1.80 - 1.91 (m, 1 H), 1.64 - 1.79 (m, 1 H), 1.45 - 1.61 (m, 1 H), 1.31 - 1.39 (d, J = 8 Hz, 3 H), 0.85 - 0.97 (m, 6 H), 0.73 - 0.81 (m, 3 H), 0.34 - 0.44 (s, 3 H).

BIOLOGICAL EXAMPLE

Compound RM461 (page 115 of FIG. 1) from WO2020132597, and compound RM351 (page 88 of FIG. 1) from WO2020132597 and compound RM44 (page 11 of FIG. 1) from WO2020132597 were cited as reference compounds for this invention.

The applicant further synthesized RM461-A and RM44-A with 2-chloro-2-fluoro- acetamide instead of chloro acetamide as the analogue of RM461 and the analogue of RM44 according to the procedure described in WO2020132597, and provided the comparison data to demonstrate the improvement and illustrate the technical problem solved herein.

Example 59

GSH Reaction Rates Glutathione (GSH) is a tripeptide found in most of the tissues, especially in high concentrations in the liver, and plays critical roles in protecting cells from oxidative damage and the toxicity of xenobiotic electrophiles, and maintaining redox homeostasis. More specifically, glutathione conjugation helps contribute to detoxification by binding electrophiles that could otherwise bind to proteins or nucleic acids, resulting in cellular damage and genetic mutations. Many potentially toxic electrophilic xenobiotics and some endogenous compounds are detoxified by conversion to the corresponding glutathione S -conjugate, which consumes inherent GSH and then diminishes detoxification effects. Some drugs and halogenated workplace/environmental contaminants are bioactivated by this mechanism.

On the other hand, conjugation between glutathione and drug molecule in extrahepatic organs as well as in the liver typically leads to the poor PK properties (notably high clearance) of the molecule, and increases its possibility for off-target reactivity (potential liability for various toxicity). Therefore Strategy to minimize the GSH metabolism is very critical. Short T 1/2 in inherent GSH reaction indicated high GSH reaction rate. Thus T 1/2 in inherent GSH reaction assay was determined for the screening of candidates.

Reference compounds and compounds of this invention potentially could form conjugation with GSH either through halogenated moieties substitution reaction or direct Michael addition reaction. This test was therefore performed to check GSH reactivity of listed compounds.

For inherent GSH reactivity determination, compounds at 1 pM were incubated at 37 °C with and without 5 mM GSH for 0, 0.5, 1, 2, 4 and 6 h in 100 mM potassium phosphate buffer at pH 7.4. At the end of the designated time points, samples were quenched with acetonitrile containing lOmM N-ethylmaleimide and an internal standard. Quenched samples were centrifuged, and supernatants were analyzed by LC-MS/MS for compound quantification. If %depletion after 6 hour incubation is less than 20%, compounds were reported as stable; if %depletion is greater than 20%, half-life values are reported.

Table 1. GSH Reaction Rates of Examples and Compounds of present invention

Above result clearly shows that reference compounds (RM461, RM351and RM44) formed conjugation with GSH causing its depletion over 6 hours while compounds of current invention maintained the stability with much less or no conjugation with GSH. Particularly, comparing reference compound RM461 and RM44 with their analogues RM461-A and RM44-A, the only difference is that the chloro acetamide was replaced by 2-chloro-2-fluoro-acetamide like the compounds of this invention, surprisingly such small change could address GSH toxicity issue as found in reference compounds.

Example 60

Human hepatocyte stability Assay

The hepatocyte stability assay measures the rate of disappearance of a compound from incubations with cryopreserved suspension hepatocytes from human. Positive controls, including Midazolam, Raloxifene and Dextromethorphan, are included in every experiment. Incubations consist of 1 pM tested compound and suspension of human hepatocytes (1 x 10 ⁶ cells/mL) in supplemented Williams’ E Medium with 10% FBS and 0.5% Penicillinstreptomycin. The hepatocyte suspension was incubated with intermittent shaking 900 rpm at 37°C, in a 5% CO2 incubator. The reaction was stopped by adding methanol containing internal standard (2 pM Tolbutamide) at 2, 10, 20, 40, 60 and 120 minutes after compound addition, depletion of the parent compound was monitored by LC-MS/MS analysis. For human data, CL_hep (mL/min/kg) >16.24 is high clearance, CL_hep (mL/min/kg) < 6.96 is low clearance. 16.24 > CL_hep (mL/min/kg) > 6.96 is medium clearance.

Table 2. Human hepatocytes stability of Examples and Compounds of present invention

Above result clearly shows that reference compounds (RM44) showed medium clearance while Example 37 of current invention maintained the low clearance in human hepatocytes stability assay. Particularly, comparing reference compound RM44 with its analogue RM44-A, the only difference is that the chloro acetamide was replaced by 2-chloro-2-fluoro-acetamide like the compounds of this invention, surprisingly such small change could achieve the clearance reduction in human hepatocytes stability assay.

Achieving low clearance is advantageous to improve in vivo performance of the compound, such as dose reduction, exposure enhancement, and half-life prolongation.

Example 61

Cell viability assay

The purpose of this cellular assay was to determine the effects of test compounds on the proliferation of human cancer cell lines NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL- 1739) cells, SW620 (ATCC-CCL-227) over a 3-day treatment period by quantifying the amount of NADPH present at endpoint using Cell Counting Kit-8.

Cells were seeded at 5,000 cells/well (NCI-H358), 2,000 cells/well (AGS) 2,000 cells/well (SW620) in 96-well assay plates (Corning-3699) and incubated overnight. On the day of the assay, diluted compounds were then added in a final concentration of 0.5% DMSO. After 72 hrs incubation, a tenth of the volume of cell counting kit 8(Dnjindo-CK04) was added into each well. Read the signal (OD450 minus OD650) using EnVision after 2 hrs incubation. IC50 was determined by fitting a 4-parameter sigmoidal concentration response model.

Compounds of this invention clearly showed overall much improved cell potency in G12C, G12D and G12V compared with reference compounds RM461 and RM351.

Table 3. Activity of Examples and Compounds of present invention in KRAS Cell viability assay

Example 62

KRAS G12C-BRAF NanoBit assay

This assay is to measure the ability of tested compounds in disruption of the KRAS G12C- BRAF complex at the cellular level, we established the NanoBit cellular assay in mammalian HEK293 (ATCC) cells.

HEK293 cells were grown and maintained using DMEM medium (Thermo Fisher Scientific) with 10% fetal bovine serum and 1% penicillin/streptomycin. Both KRAS G12C and BRAF RBD were cloned into the NanoBit vectors (BiBiT vectors system, Promega) with the orientations SmBit-KRAS G12C and BRAF RBD-LgBit, respectively, and co -transfected into HEK293 cells. Cells were then selected with 100 pg/mL Hygromycin B (10687010, Thermo Fisher) and Blasticidin (5 pg/mL) for 4 weeks to get the stable cell pool.

On the day of the assay, 75 nL of compound solution was presented in a 384-well assay plate as a 16-point 3-fold dilution starting from a final concentration of 30 pM in DMSO. Then cells were seeded at 10,000 cells/25pL/well in a 384-well plate. After 3 hours of incubation, 6pL of volume of Nano-Gio ® Live Cell Substrate (Promega) was added into each well. Monitor luminescence using ultra384 model in Envision at 20 minutes. Compounds that facilitate disruption of the KRAS G12C-BRAF RBD complex were identified as those eliciting a decrease of luminescence relative to DMSO control wells. Table 4. Activity of Examples and Compounds of present invention in KRAS G12C-BRAF

NanoBit assay

Example 63

KRAS-BRAF with CYPA (500 nM) interaction assay

In this example, TR-FRET was also used to measure the compound or compound-CYPA dependent disruption of the KRAS G12C-BRAF complex. This protocol was also used to measure disruption of KRAS G12D or KRAS G12V binding to BRAF by a compound of the invention, respectively. In assay buffer containing 25mM HEPES PH=7.4 (4-(2-hydroxyethyl)- 1 -piperazineethanesulfonic acid, Thermo, 15630080), 0.002% Tween20, 0.1% BSA, lOOmM NaCl, 5mM MgCF, 10 pM GMPPNP (Guanosine 5'-[P,y-imido]triphosphate trisodium salt hydrate, Sigma, G0635), tagless CYPA, GMPPNP loaded 6His-KRAS proteins, and GST- BRAF ^RBD were mixed in a well of a 384-well assay plate at final concentrations of 50 nM, 6.25 nM and InM, respectively. Compound was present in plate wells as a 16-point 3-fold dilution series starting at a final concentration of 10 pM and incubated for 3 hours. A mixture of MAb Anti-6His-XL665 (Cisbio, 61HISXLB) and Mab anti-GST-TB cryptate (Cisbio, 61GSTTLB)was then added at a final concentration of 6.67 nM and 0.21 nM, respectively, and the plate was incubated for an additional 1.5 hours. TR-FRET signal was read on a PHERstar FSX microplate reader (Ex320 nm, Em 665/615 nm). Compounds that facilitate disruption of the KRAS-BRAF complex were identified as those eliciting a decrease in the TR-FRET ratio relative to DMSO control wells.

Table 5. Activity of Examples and Compounds of present invention in KRAS-BRAF with CYPA (500 nM) interaction assay

Example 64 pERK inhibition assay

This assay is to measure the ability of test compounds in inhibiting the phosphorylation of ERK, the downstream signaling of KRAS G12C in NCI-H358 cells, KRAS G12D in AGS cells, and KRAS G12V in SW620. NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL-1739) cells, SW620 (ATCC-CCL-227) cells were all grown and maintained using RPMI-1640 medium (Thermo Fisher Scientific) with 10% fetal bovine serum and 1% penicillin/streptomycin. On the day prior to compound addition, cells were plated in tissue culture -treated 96 well plates (Corning-3699) at a density of 30,000 cell/well, 20,000 cell/well, 30,000 cell/well for NCI-H358, AGS and SW620 respectively, and allowed for attachment overnight. Diluted compounds were then added in a final concentration of 0.5% DMSO. After 4 hours of incubation, the medium was removed, 100 pL of 4% formaldehyde was added, and the assay plates were incubated at room temperature for 20 minutes. The plates were then washed once with phosphate buffered saline (PBS), and permeabilized with 100 pL of chilled methanol for 10 minutes. Non-specific antibody binding to the plates was blocked using 50 pL IX BSA blocking buffer (Thermo -37520, 10-fold dilution by Phosphate-Buffered Saline Tween (PBST) for at least 1 hour at room temperature.

The amount of phosphor-ERK was determined using an antibody specific for phosphorylated form of ERK. Primary antibody (pERK, CST-4370, Cell Signaling Technology) was diluted 1:300 in blocking buffer, with 50 pL aliquoted to each well, and incubated overnight at 4 °C. Cells was washed five times for 5 minutes with PBST. Secondary antibody (HRP-linked anti-rabbit IgG, CST-7074, Cell Signaling Technology) was diluted 1:1000 in blocking buffer, and 50 pL was added to each well and incubated 1-2 hrs at room temperature. Cells was washed 5 times for 5 minutes with PBST, lOOpL TMB ELISA substrate (abcam-abl71523) were added and gently shake for 20 minutes. 50pL stop solution (abcam-abl71529) were added, and then read the signal (OD450) by EnVision.

IC50 was determined by fitting a 4-parameter sigmoidal concentration response model.

Table 6. Activity of Examples and Compounds of present invention in KRAS pERK inhibition assay