Macrocyclic compounds useful as KRAS inhibitors

Abstract

The present invention relates to compounds of formula (I) # imgabs0 # wherein R1 to R6, A1 and A2 are as described herein, and pharmaceutically acceptable salts thereof, as well as compositions comprising the compounds and methods of using the compounds as KRAS inhibitors.

Description

[0001] The present invention relates to organic compounds useful for treatment and / or prevention in mammals, and in particular to the inhibition of KRAS mutants useful for the treatment of cancer. Technical Field

[0002] RAS is one of the best-known oncogenes. About 30% of human cancers contain mutations in the three most significant members (KRAS, HRAS, and NRAS), making them the most common oncogenic drivers. KRAS mutations are often associated with poor prognosis, especially in colorectal cancer, pancreatic cancer, and lung cancer. As the most frequently mutated RAS subtype, KRAS has been extensively studied in the past few years. Among the most common KRAS alleles (including G12D, G12V, G12C, G13D, G12R, G12A, G12S, Q61H, etc.), G12C, G12D, and G12V account for more than half of all K-RAS-driven cancers (including colorectal cancer (CRC), pancreatic ductal adenocarcinoma (PDAC), and lung adenocarcinoma (LUAD)). Notably, among all KRAS-altered cancers (ovarian, esophageal and gastric, uterine), KRAS wild-type amplification was also found in approximately 7%, ranking among the top alterations.

[0003] All RAS proteins belong to the protein family of small GTPases that hydrolyze GTP to GDP. KRAS is structurally divided into an effector-binding lobe, followed by an allosteric lobe and a carboxyl-terminal region responsible for membrane anchoring. The effector lobe includes the P-loop, switch I, and switch II regions. The switch I / II loop plays a key role in KRAS downstream signaling by mediating protein-protein interactions with effector proteins, such as RAF in the mitogen-activated protein kinase (MAPK) pathway or PI3K in the phosphatidylinositol 3-kinase (PI3K) / protein kinase B (AKT) pathway.

[0004] The KRAS protein switches between an inactive form and 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 seven son homolog 1 (SOS1) or GTPase activating proteins (GAPs) involved in catalyzing the exchange of GDP for GTP, thereby enhancing intrinsic GTPase activity or accelerating RAS-mediated GTP hydrolysis. In response to extracellular stimuli, inactive RAS-GDP is converted to active RAS-GTP, which directly binds to the RAF RAS binding domain (RAF RBD) binds to the RAF family of kinases, thereby recruiting the RAF kinase family from the cytoplasm to the cell membrane, where they dimerize and become active. The activated RAF then undergoes a series of phosphorylation reactions on its downstream mitogen-activated protein kinase (MEK) and extracellular signal-regulated kinase (ERK), and propagates growth signals. Among the RAF protein kinase family (three known subtypes ARAF, BRAF, CRAF / RAF1), BRAF is the most frequently mutated and remains the most potent activator of MEK. Although individual RAS and RAF family members exhibit different binding preferences, all RAFs have a conserved RBD for forward transmission of MAPK signaling, which is frequently used to characterize KRAS inhibition (such as KRAS-BRAF in this article). RBD For KRAS, mutations at positions 12, 13, 61, and 146 lead to a shift to the active KRAS form by impairing nucleotide hydrolysis or activating nucleotide exchange, thereby causing overactivation of the MAPK pathway that causes tumorigenesis.

[0005] Despite its widely recognized importance in cancer malignancies, sustained efforts in the past have failed to develop approved therapies for KRAS mutant cancers until recently, when the first selective drug, AMG510, was rapidly approved as a second-line treatment for KRAS G12C-driven non-small cell lung cancer (NSCLC). However, clinically acquired resistance to KRAS G12C inhibitors emerges dramatically as the disease progresses after approximately 6 months of treatment. All mutations converge to reactivate RAS-MAPK signaling, with secondary RAS mutants observed at oncogenic hotspots (e.g., G12 / G13 / Q61) and within the switch II pocket (e.g., H95, R68, and Y96); in addition, more than 85% of all cancers driven by KRAS mutant or wild-type amplification still lack novel agents. In summary, both the myriad escape mechanisms and the variety of oncogenic alleles highlight the urgent medical need for additional KRAS therapies. Therefore, we have developed oral compounds that target and inhibit KRAS alleles for the treatment of KRAS mutant-driven cancers. Summary of the invention

[0006] The present invention relates to novel compounds having formula (I),

[0007]

[0008] in

[0009] R 1 is 3-oxabicyclo[3.1.0]hexane, C 1-6 Alkyl C 3-7 Cycloalkyl, cyano C 3-7 Cycloalkyl or halogenated C1-6 Alkyl C 3-7 Cycloalkyl;

[0010] R 2 is H or halogen;

[0011] R 3 is H or halogen;

[0012] R 4 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0013] R 5 C 1-6 Alkoxy C 1-6 alkyl;

[0014] R 6 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl;

[0015] A 1 is thiazolylidene;

[0016] A 2 C 1-6 Alkylene;

[0017] The condition is R 2 and R 3 Not at the same time H;

[0018] or a pharmaceutically acceptable salt thereof.

[0019] The invention also relates to their manufacture, to medicaments based on the compounds according to the invention and their production and to the use of the compounds of formula (I) or (Ia) as KRAS inhibitors.

[0020] The compounds of formula (I) or (Ia) show good KRAS inhibition on G12C and G12V. In another embodiment, the compounds of the present invention show excellent cancer cell inhibition and human hepatocyte stability. In addition, the compounds of formula (I) or (Ia) also show good or improved cytotoxicity and solubility characteristics. In addition, compared with the reference compound, the compounds of the present invention have good pharmacokinetic properties. DETAILED DESCRIPTION

[0021] definition

[0022] The term "C 1-6 "Alkyl" means a saturated, straight-chain or branched alkyl group containing 1 to 6, especially 1 to 4 carbon atoms, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, etc. In particular, "C 1-6"Alkyl" groups are methyl, ethyl and n-propyl.

[0023] The term "C 1-6 "Alkoxy" means C 1-6 Alkyl-O-.

[0024] The term “(C 1-6 ")Alkylene" means a straight 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. 1-6 ) Examples of the alkylene group include methylene, ethylene, propylene, 2-methylpropylene, butylene, 2-ethylbutylene, pentylene, hexylene.

[0025] The terms "halogen" and "halo" are used interchangeably herein to refer to fluorine, chlorine, bromine or iodine.

[0026] The term "halogenated C 1-6 "Alkyl" means C 1-6 Alkyl groups, where C 1-6 At least one hydrogen atom of the alkyl group has been replaced by identical or different halogen atoms, in particular fluorine atoms. Examples of haloalkyl groups include monofluoro-, difluoro- or trifluoro-methyl, -ethyl or -propyl, for example 3,3,3-trifluoropropyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, fluoromethyl or trifluoromethyl.

[0027] The term "C 3-7 "Cycloalkyl" means a monovalent saturated monocyclic or bicyclic hydrocarbon radical of 3 to 7 ring carbon atoms. Bicyclic means consisting of two saturated carbocyclic rings having one or two common carbon atoms. Examples of monocyclic cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. Examples of bicyclic cycloalkyl are bicyclo[1.1.0]butyl, bicyclo[2.2.1]heptyl, bicyclo[1.1.1]pentanyl or bicyclo[2.2.2]octyl.

[0028] The term "thiazolylene" refers to a divalent thiazolyl group.

[0029] The term "oxo" refers to a divalent oxygen atom =0.

[0030] The term "2,2-propylene" means

[0031] The term "blocking group" means a group that selectively blocks a reactive site in a multifunctional compound so that a chemical reaction occurs selectively on another unprotected reactive site that is usually associated therewith in synthetic chemistry. The blocking group can be removed at an appropriate time point. Exemplary blocking groups are amino-protecting groups, carboxyl-protecting groups or hydroxyl-protecting groups.

[0032] It will be appreciated by those skilled in the art that the following structures of the compounds of formula (I) and (I') are identical, especially with respect to the chiral center:

[0033]

[0034] The term "pharmaceutically acceptable salts" refers to salts that are not biologically or otherwise undesirable. "Pharmaceutically acceptable salts" include both acid addition salts and base addition salts.

[0035] "Pharmaceutically acceptable acid addition salts" refer to those pharmaceutically acceptable salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and the like, and organic acids, which may be selected from aliphatic, alicyclic, aromatic, aromatic aliphatic, heterocyclic, formic and sulfonic acid organic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid, and the like.

[0036] The term "pharmaceutical base addition salt" refers to those pharmaceutical salts formed with organic or inorganic bases. Examples of acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese and aluminum salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include primary amines, secondary amines, 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, tromethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine and polyamine resins).

[0037] The term "pharmaceutically active metabolite" refers to a pharmacologically active product produced by the metabolism of a specific compound or its salt in the body. After entering the human body, most drugs are substrates for chemical reactions that may change their physical properties and biological effects. These metabolic transformations, which usually affect the polarity of the compounds of the present invention, change the way the drug is distributed in the body and excreted from the body. However, in some cases, drug metabolism is necessary for therapeutic effects.

[0038] The term "therapeutically effective amount" refers to an amount of a compound or molecule of the invention that, when administered to a subject, (i) treats or prevents a particular disease, disorder or condition, (ii) reduces, ameliorates or eliminates one or more symptoms of a particular disease, disorder or condition, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, disorder or condition as described herein. The therapeutically effective amount depends on the compound, the disease state being treated, the severity of the disease being treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending medical or veterinary doctor and other factors.

[0039] The term "pharmaceutical composition" means a mixture or solution comprising a therapeutically effective amount of an active drug ingredient together with a pharmaceutically acceptable excipient for administration to a mammal (eg, a human) in need thereof.

[0040] The terms "pharmaceutically acceptable excipient," "pharmaceutically acceptable carrier," and "therapeutically inert excipient" are used interchangeably and refer to any pharmaceutical ingredient in a pharmaceutical composition that is not therapeutically active and is non-toxic to the subject to which it is administered, such as a disintegrant, binder, filler, solvent, buffer, tonicity agent, stabilizer, antioxidant, surfactant, carrier, diluent, or lubricant used to formulate a drug product.

[0041] KRAS inhibitors

[0042] The present invention relates to (i) a compound of formula (I),

[0043]

[0044] in

[0045] R 1 is 3-oxabicyclo[3.1.0]hexane, C 1-6 Alkyl C 3-7 Cycloalkyl, cyano C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl;

[0046] R 2 is H or halogen;

[0047] R 3 is H or halogen;

[0048] R 4 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0049] R 5 C 1-6 Alkoxy C 1-6 alkyl;

[0050] R 6 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl;

[0051] A 1 is thiazolylidene;

[0052] A 2 C 1-6 Alkylene;

[0053] The condition is R 2 and R 3 Not at the same time H;

[0054] or a pharmaceutically acceptable salt thereof.

[0055] Another embodiment of the present invention is (ii) a compound of formula (Ia),

[0056]

[0057] in

[0058] R 1 is 3-oxabicyclo[3.1.0]hexane, C 1-6 Alkyl C 3-7 Cycloalkyl, cyano C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl;

[0059] R 2 is H or halogen;

[0060] R 3 is H or halogen;

[0061] R 4 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0062] R 5 C 1-6 Alkoxy C 1-6 alkyl;

[0063] R 6 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl;

[0064] A 1 is thiazolylidene;

[0065] A 2 C 1-6 Alkylene;

[0066] The condition is R2 and R 3 Not at the same time H;

[0067] or a pharmaceutically acceptable salt thereof.

[0068] A further embodiment of the 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 C 1-6 Alkyl C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl.

[0069] A further embodiment of the present invention is (iv) a compound of formula (I) or (Ia) or a pharmaceutically acceptable salt thereof according to any one of (i) to (iii), wherein R 1 It is methylcyclopropyl or (difluoromethyl)cyclopropyl.

[0070] A further embodiment of the present invention is (v) a compound of formula (I) or (Ia) according to any one of (i) to (iv), wherein R 2 is H or fluorine.

[0071] A further embodiment of the present invention is (vi) a compound of formula (I) or (Ia) according to any one of (i) to (v), or a pharmaceutically acceptable salt thereof, wherein R 3 is H or fluorine.

[0072] A further embodiment of the present invention is (vii') a compound of formula (I) or (Ia) according to any one of (i) to (vi), or a pharmaceutically acceptable salt thereof, wherein R 4 It is ethyl or 2,2,2-trifluoroethyl.

[0073] A further embodiment of the present invention is (viii') a compound of formula (I) or (Ia) according to any one of (i) to (vii), or a pharmaceutically acceptable salt thereof, wherein R 5 It is 1-methoxyethyl.

[0074] A further embodiment of the present invention is (ix) a compound of formula (I) or (Ia) according to any one of (i) to (viii), or a pharmaceutically acceptable salt thereof, wherein R 6 is morpholinyl or C 1-6 Alkylpiperazinyl.

[0075] A further embodiment of the present invention is (x) a compound of formula (I) or (Ia) according to any one of (i) to (ix), or a pharmaceutically acceptable salt thereof, wherein R 6 It is morpholinyl or 4-methylpiperazin-1-yl.

[0076] A further embodiment of the present invention is (xi) a compound of formula (I) or (Ia) according to any one of (i) to (x), or a pharmaceutically acceptable salt thereof, wherein A 1 for Where bond "a" is attached to the indole ring.

[0077] A further embodiment of the present invention is (xii) a compound of formula (I) or (Ia) according to any one of (i) to (xi), or a pharmaceutically acceptable salt thereof, wherein A 2 It is 2,2-propylene.

[0078] Another embodiment of the present invention is (xiii) a compound of formula (I) or formula (Ia) according to (i) or (ii), wherein

[0079] R 1 C 1-6 Alkyl C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl;

[0080] R 2 is H or halogen;

[0081] R 3 is H or halogen;

[0082] R 4 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0083] R 5 C 1-6 Alkoxy C 1-6 alkyl;

[0084] R 6 is morpholinyl or C 1-6 Alkylpiperazinyl;

[0085] A 1 for wherein bond "a" is attached to the indole ring;

[0086] A 2 C 1-6 Alkylene;

[0087] The condition is R 2 and R 3 Not at the same time H;

[0088] or a pharmaceutically acceptable salt thereof.

[0089] Another embodiment of the present invention is (xiv) a compound of formula (I) or formula (Ia) according to (xiii), wherein

[0090] R 1 is 2-methylcyclopropyl or 2-(difluoromethyl)cyclopropyl;

[0091] R 2 is H or fluorine;

[0092] R 3 is H or fluorine;

[0093] R 4 is ethyl or 2,2,2-trifluoroethyl;

[0094] R 5 is (1S)-1-methoxyethyl;

[0095] R 6 is morpholinyl or 4-methylpiperazin-1-yl;

[0096] A 1 for wherein bond "a" is attached to the indole ring;

[0097] A 2 is 2,2-propylene;

[0098] The condition is R 2 and R 3 Not at the same time H;

[0099] or a pharmaceutically acceptable salt thereof.

[0100] Another embodiment of the present invention is (xv) a compound of formula (I) or (Ia) selected from the following compounds:

[0101] (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide;

[0102] (1R,5S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-oxabicyclo[3.1.0]hexane-6-carboxamide;

[0103] (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide;

[0104] (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide;

[0105] (1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide;

[0106] (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide;

[0107] (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide;

[0108] (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide;

[0109] (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide;

[0110] (1S,2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide;

[0111] (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide; and

[0112] (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide;

[0113] or a pharmaceutically acceptable salt thereof.

[0114] Another embodiment of the present invention relates to (xvi) a method for preparing a compound according to any one of (i) to (xv), the method comprising the following steps:

[0115] a) making a compound of formula (II),

[0116] With acid (III), A coupling reaction is carried out in the presence of a coupling reagent and a base to form a compound of formula (I);

[0117] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , A 1 and A 2 As defined in any one of (i) to (xiv); the coupling reagent is T 3 P, HATU, PyBOP or EDCI / HOBt; the base is TEA, DIEPA or DMAP.

[0118] Another embodiment of the present invention is (xvii) a compound or a pharmaceutically acceptable salt according to any one of (i) to (xv) for use as a therapeutically active substance.

[0119] Another embodiment of the present invention is (xviii) a pharmaceutical composition comprising the compound according to any one of (i) to (xv) and a pharmaceutically acceptable excipient.

[0120] Another embodiment of the present invention is (xix) use of a compound according to any one of (i) to (xv) for treating a KRAS G12C protein-related disease.

[0121] Another embodiment of the present invention is (xx) use of a compound according to any one of (i) to (xv) for treating KRAS G12C, G12D and G12V protein related diseases.

[0122] Another embodiment of the present invention is (xxi) use of a compound according to any one of (i) to (xv) for inhibiting the interaction of RAS with a downstream effector, wherein the downstream effector is RAF and PI3K.

[0123] Another embodiment of the present invention is (xxii) use of a compound according to any one of (i) to (xv) for inhibiting propagated oncogenic MAPK and PI3K signaling.

[0124] Another embodiment of the present invention is (xxiii) use of a compound according to any one of (i) to (xv) for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma ovarian cancer and endometrial cancer.

[0125] Another embodiment of the present invention is (xxiv) use of a compound according to any one of (i) to (xv) for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0126] Another embodiment of the present invention is (xxv) a compound or a pharmaceutically acceptable salt according to any one of (i) to (xv), for use in treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0127] Another embodiment of the present invention is (xxvi) use of a compound according to any one of (i) to (xv) for the preparation of a medicament for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0128] Another embodiment of the present invention is (xxvii) a method for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer, the method comprising administering a therapeutically effective amount of a compound as defined in any one of (i) to (xv).

[0129] Another embodiment of the present invention is (xxviii) a compound or a pharmaceutically acceptable salt according to any one of (i) to (xv), which is prepared according to the method of (xvi).

[0130] Pharmaceutical compositions and administration

[0131] Another embodiment provides a pharmaceutical composition or medicine containing the compounds of the invention and a therapeutic inert carrier, diluent or excipient, and methods for preparing such compositions and medicines using the compounds of the invention. In one example, the compound of formula (I) can be formulated as a galenical administration form by mixing with a physiologically acceptable carrier (i.e., a carrier that is nontoxic to the recipient at the dosage and concentration used) at ambient temperature at an appropriate pH and desired purity. The pH of the formulation depends primarily on the specific use and concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, the compound of formula (I) is formulated in an acetate buffer at pH 5. In another embodiment, the compound of formula (I) is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0132] The composition is formulated, metered, and administered in a manner consistent with good medical practice. Factors to be considered in this case include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the delivery site of the agent, the method of administration, the timing of administration, and other factors known to practitioners. The "effective amount" of the compound to be administered will be affected by such considerations and is the minimum amount necessary to inhibit the interaction of mutant RAS (e.g., KRAS G12C) with RAF, thereby blocking oncogenic MAPK signaling. For example, the amount may be lower than an amount that is toxic to normal cells or mammals as a whole.

[0133] In one example, the pharmaceutically effective amount of the compound of the present invention administered parenterally per dose will be in the range of about 0.1 to 1000 mg / kg patient body weight per day, alternatively about 0.1 to 1000 mg / kg patient body weight, typically an initial range of 0.3 to 15 mg / kg / day of the compound used. In another embodiment, oral unit dosage forms such as tablets and capsules preferably contain about 1 to about 1000 mg of the compound of the present invention.

[0134] The compounds of the present invention can 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 necessary for local treatment) intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

[0135] The compounds of the present invention can be administered in any convenient administration form, for example, tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain conventional components of pharmaceutical preparations, for example, diluents, carriers, pH adjusters, sweeteners, fillers and other active agents.

[0136] Typical formulations are prepared by mixing the compounds of the invention with carriers or excipients. Suitable carriers and excipients are well known to those skilled in the art and are described in, for example, 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: Science and Practice of Pharmacy .Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients .Chicago, Pharmaceutical Press, 2005. The formulation may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavoring agents, flavoring agents, diluents and other known additives to provide an aesthetically pleasing display of a drug (e.g., a compound of the present invention or a pharmaceutical composition thereof) or to facilitate the preparation of a pharmaceutical product (e.g., a drug).

[0137] An example of a suitable oral dosage form is a tablet containing about 1 to 1000 mg of a compound of the present invention and about 1 to 1000 mg of anhydrous lactose, about 1 to 1000 mg of cross-linked sodium carboxymethylcellulose, about 1 to 1000 mg of polyvinyl pyrrolidone (PVP) K30 and about 1 to 1000 mg of magnesium stearate. The powdered ingredients are first mixed together and then mixed with the PVP solution. The resulting composition can be dried, granulated, mixed with magnesium stearate and compressed into tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound of the present invention (e.g., 5 mg to 400 mg) in a suitable buffer solution (e.g., phosphate buffer) and adding a penetration enhancer (e.g., a salt such as sodium chloride) if necessary. The solution can be filtered, for example, using a 0.2 micron filter to remove impurities and contaminants.

[0138] Thus, embodiments include a pharmaceutical composition comprising a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof. Further embodiments include a pharmaceutical composition comprising a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0139] Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in treating a mutant KRAS driven cancer. Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in treating a mutant KRAS driven cancer.

[0140] The following compositions A and B illustrate typical compositions of the present invention but are intended to be representative thereof only.

[0141] Composition A

[0142] The compounds of the invention can be used as active ingredients in a manner known per se to produce tablets of the following composition:

[0143]

[0144]

[0145] Composition B

[0146] The compounds of the invention can be used as active ingredients in a manner known per se to produce capsules of the following composition:

[0147]

[0148] Indications and treatments

[0149] The compounds of the present invention induce new binding pockets in KRAS by driving the formation of a high-affinity tri-complex between KRAS protein and widely expressed cyclophilin A (CYPA), and these compounds inhibit the interaction of KRAS with downstream effectors (such as RAF and PI3K). Therefore, the compounds of the present invention can be used to inhibit the propagation of oncogenic MAPK and PI3K signal transduction, reducing cell (especially cancer cell) proliferation. The compounds of the present invention can be used to terminate RAS signal transduction in cells of pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma ovarian cancer, endometrial cancer, etc. driven by RAS mutants (especially KRAS mutations). Alternatively, the compounds of the present invention can be used to terminate RAS signal transduction in malignant solid tumors, wherein the carcinogenic effect of KRAS mutations is enhanced by the dysregulation or mutation of effector pathways such as signal transduction driven by MAPK, PI3K-AKT-mTOR (mammalian rapamycin target), and these compounds are used for targeted therapies of pancreatic cancer, colorectal cancer, non-small cell lung cancer, etc.

[0150] 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.

[0151] synthesis

[0152] The compounds of the present invention may be prepared by any conventional method. Suitable methods for synthesizing these compounds and their starting materials are provided in the following schemes and examples. Unless otherwise indicated, all substituents, especially R 1 To R 6 , A 1 and A 2 As defined above. Furthermore, unless otherwise specifically indicated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to one of ordinary skill in the art of organic chemistry.

[0153] The general synthetic route for preparing compounds of formula (I) is shown below.

[0154] Solution 1

[0155]

[0156] Compounds of formula II are synthesized according to the procedures described in Intermediates A to I. Compounds of formula (I) can be prepared by reacting the mixture with a coupling reagent such as TEA, DIEPA and DMAP in the presence of a base such as TEA, DIEPA and DMAP. 3 The obtained product is obtained by coupling reaction between the acid (III) and the compound of formula (II) using P, HATU, PyBOP and EDCI / HOBt.

[0157] The compounds of the present invention can be obtained as diastereomers or mixtures of enantiomers, which can be separated by methods well known in the art, for example, (chiral) HPLC or SFC. In another embodiment, compounds of formula (I) can be obtained according to the above scheme using the corresponding chiral starting materials.

[0158] The present invention also relates to a method for preparing a compound of formula (I), the method comprising the following steps:

[0159] a) making a compound of formula (II),

[0160] With acid (III), A coupling reaction is carried out in the presence of a coupling reagent and a base to form a compound of formula (I);

[0161] in

[0162] In step a), the coupling reagent may be, for example, T 3P, HATU, PyBOP or EDCI / HOBt; the base can be, for example, TEA, DIEPA or DMAP.

[0163] The compounds of formula (I) or (Ia) produced according to the above-described process are also an object of the present invention.

[0164] Examples

[0165] The present invention will be more fully understood by reference to the following examples. However, they should not be construed as limiting the scope of the present invention.

[0166] abbreviation

[0167] The present invention will be more fully understood by reference to the following examples. However, they should not be construed as limiting the scope of the present invention.

[0168] The abbreviations used in this article are as follows:

[0169] ACN Acetonitrile

[0170] aq. water-based

[0171] Boc-N-Me-Val-OH N-(tert-Butoxycarbonyl)-N-methyl-L-valine

[0172] (Boc) 2 O Di-tert-butyl dicarbonate

[0173] (R)-binap (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl

[0174] CDCl 3 : Deuterated chloroform

[0175] CD 3 OD: Deuterated methanol

[0176] COMU (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbonium hexafluorophosphate

[0177] DIEPA: N,N-diethylpropylamine

[0178] DIBAL-H Diisobutylaluminum hydride

[0179] DMAP: 4-dimethylaminopyridine

[0180] DMF: Dimethylformamide

[0181] DMP 1,1,1-tri(acetoxy)-1,1-dihydro-1,2-benzooxyphenol-3-(1H)-one

[0182] DMSO: dimethyl sulfoxide

[0183] EDCI: N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride

[0184] EtOAc or EA: Ethyl acetate

[0185] FRET Fluorescence Resonance Energy Transfer

[0186] HATU: (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate)

[0187] hr: hours

[0188] HPLC: High Performance Liquid Chromatography

[0189] HOBt: N-Hydroxybenzotriazole

[0190] H-VAL-OTBU HCl (S)-tert-Butyl 2-amino-3-methylbutyrate hydrochloride

[0191] [Ir(OMe)(COD)] 2 (1,5-cyclooctadiene)(methoxy)iridium(I) dimer

[0192] LDA Lithium diisopropylamide

[0193] MS: (ESI): Mass spectrometry (electrospray ionization)

[0194] min

[0195] MTBE Methyl tert-butyl ether

[0196] NMM N-Methylmorpholine

[0197] NMR: Nuclear Magnetic Resonance

[0198] NMO 4-Methylmorpholine N-oxide

[0199] obsd. Observed value

[0200] Pd(dppf)Cl 2 [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II) chloride

[0201] Pd(dtbpf)Cl 2 [1,1'-Bis(di-tert-butylphosphino)ferrocene]palladium(II) dichloride

[0202] prep-HPLC Preparative High Performance Liquid Chromatography

[0203] PyBOP: Benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate

[0204] RT or rt: room temperature

[0205] sat. saturated

[0206] SFC Supercritical Fluid Chromatography

[0207] TEA: Triethylamine

[0208] TFA: trifluoroacetic acid

[0209] THF: Tetrahydrofuran

[0210] TEA: Trimethylamine

[0211] TMEDA Tetramethylethylenediamine

[0212] TMSCF 3 Trifluoromethyltrimethylsilane

[0213] T 3 P: Propylphosphonic anhydride

[0214] General experimental conditions

[0215] Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and Quad 12 / 25 Cartridge module. ii) ISCO combi-flash chromatograph. Silica gel brand and pore size: i) KP-SIL Particle size: 40-60 μm; ii) CAS registration number: silica gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Ocean Chemical Co., Ltd., pore size: 200-300 or 300-400.

[0216] Intermediates and final compounds were purified by preparative HPLC on a reverse phase column using XBridge TM Prep-C18 (5μm, OBDTM 30×100mm) column, SunFire TM Prep-C18(5μm,OBD TM30×100mm) chromatographic column, Phenomenex Synergi-C18 (10μm, 25×150mm) or Phenomenex Gemini-C18 (10μm, 25×150mm). 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% HCl in water; acetonitrile and 0.075% TFA in water; or acetonitrile and water).

[0217] For SFC chiral separation, the intermediates were separated by chiral columns (Daicel chiralpak IC, 5 μm, 30 × 250 mm), AS (10 μm, 30 × 250 mm) or AD (10 μm, 30 × 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 solution) or CO 2 and MeOH (0.1% NH 3 ·H 2 O in MeOH), back pressure 100 bar, UV detection at 254 or 220 nm.

[0218] Using LC / MS (Waters TM The LC / MS spectra of the compounds were obtained using an Agilent Alliance 2795-Micromass ZQ, Shimadzu Alliance 2020-Micromass ZQ, or Agilent Alliance 6110-Micromass ZQ. The LC / MS conditions were as follows (run time 3 or 1.5 min):

[0219] Acidic conditions I: A: 0.1% TFA in HO 2 O solution; B: 0.1% TFA solution in acetonitrile;

[0220] Acidic conditions II: A: 0.0375% TFA in HO 2 O solution; B: 0.01875% TFA solution in acetonitrile;

[0221] Alkaline conditions I: A: 0.1% NH 3 ·H 2 O in H 2 O solution; B: acetonitrile;

[0222] Alkaline conditions II: A: 0.025% NH 3 ·H 2 O in H 2 O solution; B: acetonitrile;

[0223] Neutral conditions: A:H 2 O; B: acetonitrile.

[0224] Mass spectra (MS): Usually only ions representing the parent mass are reported, and unless otherwise stated, the mass ion quoted is the positive mass ion (MH) + .

[0225] NMR spectra were obtained using a Bruker Avance 400 MHz or 500 MHz.

[0226] Microwave-assisted reactions were performed in a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were performed under argon or nitrogen atmosphere. Unless otherwise stated, reagents were purchased from commercial suppliers as received without further purification.

[0227] Preparation Example

[0228] Preparation of intermediates

[0229] Intermediate A1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-methyl-piperazine

[0230]

[0231] The title intermediate A was prepared according to the following scheme:

[0232]

[0233] Step 1: Preparation of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (Compound A2)

[0234] To a solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (Compound A1, 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-tert-butyl-2,2'-bipyridine (372.7 mg, 1.39 mmol) and [Ir(OMe)(COD)] 2 (306.3 mg, 0.460 mmol). The mixture was stirred at N 2 The mixture was stirred at 75 ° C for 16 hours. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (EA / PE: 0-20%) to give 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (Compound A2, 2.4 g) as a yellow oil. 1 H NMR (400MHz, CDCl3) δppm 8.91 (d, J = 1.4Hz, 1H), 8.21 (d, J = 1.4Hz, 1H), 4.95 (q, J = 6.5Hz, 1H), 3.30 (s, 3H), 1.49 (d, J = 6.5Hz, 3H), 1.35 (s, 12H).

[0235] Step 2: Preparation of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3)

[0236] To a solution of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)pyridine (Compound A2, 2.5 g, 7.3 mmol) in ACN (40 mL) was added N-iodosuccinimide (4.1 g, 18.27 mmol). 2 The mixture was stirred at 90°C for 40 hours under protection. 2 SO 3 The solution (40mL) was quenched, and the reaction mixture was extracted with EtOAc (30mL, twice). The combined organic layer was washed with brine (50mL), filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA / PE: 0-20%) to obtain 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3, 660mg) as a yellow oil. MS calculated value 342 (MH + ); measured value 341.8 (MH + ).

[0237] Step 3: Preparation of benzyl 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylate (Compound A5)

[0238] To a solution of 3-bromo-5-iodo-2-[(1S)-1-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) was added cesium carbonate (1.6 g, 4.83 mmol), (R)-BINAP (60.1 mg, 0.1 mmol) and palladium (II) acetate (43.3 mg, 0.19 mmol). The mixture was stirred under N 2 The mixture was stirred at 100 ° C for 12 hours. The mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA / PE: 0-50%) to give 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound A5, 740 mg) as a yellow solid. MS calculated value 434.1 (MH + ); measured value 434.1 (MH + ).

[0239] Step 4: Preparation of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-methyl-piperazine (Intermediate A)

[0240] To a solution of benzyl 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylate (Compound A5, 740 mg, 1.7 mmol) and bis(pinacolato)diboron (519.2 mg, 2.04 mmol) in toluene (12 mL) was added KOAc (418.0 mg, 4.26 mmol) and Pd(dppf)Cl 2 (124.7 mg, 0.170 mmol). The reaction mixture was stirred at N 2 The mixture was stirred at 90 ° C for 12 hours under protection. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by silica gel column to give 1- [6- [(1S) -1-methoxyethyl] -5- (4,4,5,5-tetramethyl -1,3,2- dioxaborolan -2-yl) -3-pyridyl] -4-methyl-piperazine (Intermediate A, 470 mg) as a brown solid. MS calculated value 482.3 (MH + ); measured value 482.2 (MH + ).

[0241] Intermediate B

[0242] (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester

[0243]

[0244] Intermediate B was prepared according to the following scheme:

[0245]

[0246] Step 1: Preparation of (4-bromothiazol-2-yl)methanol (Compound B2)

[0247] At 0 ° C, sodium borohydride (1.7 g, 46.87 mmol) was added to a solution of 4-bromothiazole-2-carboxaldehyde (compound B1, 6.0 g, 31.25 mmol) in methanol (70 mL). 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 with ethyl acetate (200 mL, three times). The combined organic phases were washed with brine (150 mL, twice), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (4-bromothiazol-2-yl)methanol (compound B2, 6 g) as a colorless oil.

[0248] Step 2: Preparation of 4-bromo-2-(bromomethyl)thiazole (Compound B3)

[0249] To a solution of (4-bromothiazol-2-yl)methanol (Compound B2, 6.0 g, 30.92 mmol) in DCM (80 mL) was added CBr at 0 °C. 4 (15.4 g, 46.38 mmol) and triphenylphosphine (12.1 g, 46.38 mmol). After stirring 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 eluting with ethyl acetate (0-10%) in petroleum ether to give (4-bromothiazol-2-yl)methanol (Compound B3, 6.0 g) as a yellow oil. MS calculated value 255.9 (MH + ); measured value 255.9 (MH + ).

[0250] Step 3: Preparation of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5)

[0251] To a mixture of (R)-2,5-dihydro-3,6-dimethoxy-2-isopropylpyrazine (Compound B4, 4.3 g, 23.45 mmol) in THF (60 mL) was slowly added n-butyllithium (10 mL, 25.22 mmol, 2.5 M) at -78°C. After addition, the mixture was stirred at -78°C for 0.5 h. 4-Bromo-2-(bromomethyl)thiazole (Compound B3, 5.4 g, 21.02 mmol) was added to the above mixture at -78°C and the mixture was stirred for another 1 h. The reaction was heated with saturated NH 4 Cl solution (100mL) is quenched, and the reaction mixture is extracted with EtOAc (100mL, twice). The combined organic layer is washed with brine (150mL), dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated under vacuum. The residue is purified by reverse phase chromatography to obtain 4-bromo-2-[[(2S, 5R) -5- isopropyl -3,6- dimethoxy -2,5- dihydropyrazine -2- bases] methyl] thiazole (compound B5, 3.6g) as a yellow oil. MS calculated value 360 ​​(MH + ); measured value 359.9 (MH + ).

[0252] Step 4: Preparation of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (Compound B6)

[0253] To a solution of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5, 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 h. The mixture was washed with saturated NaHCO 3 The solution was basified until pH = 8. The mixture was extracted twice with EtOAc (80 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoic acid methyl ester (Compound B6, 3.1 g) as a yellow oil. MS calculated value 264.9 (MH + ); measured value 264.9 (MH + ).

[0254] Step 5: Preparation of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoate (Compound B7)

[0255] To a solution of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (compound B6, 3.1 g, 11.69 mmol) in DCM (40 mL) was added triethylamine (2.9 g, 29.23 mmol) and (Boc 2 O (3.8 g, 17.54 mmol). After stirring at 30 ° C for 12 hours, the mixture was concentrated under vacuum. The residue was purified by silica gel column eluted with ethyl acetate (0-30%) in petroleum ether to give (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid methyl ester (Compound B7, 3.2 g) as a yellow oil. MS calculated value 387 (MNa + ); measured value 386.9 (MNa + ).

[0256] Step 6: Preparation of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (Compound B8)

[0257] To a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid methyl ester (Compound B7, 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 stirring at 25 ° C for 1 hour, the reaction mixture was acidified with 1M HCl solution until pH = 5. The mixture was extracted twice with EtOAc (40 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (Compound B8, 3.1 g) as a yellow oil. MS calculated value 373 (MNa + ); measured value 372.9 (MNa + ).

[0258] Step 7: Preparation of methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (Intermediate B)

[0259] To a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (compound B8, 3.1 g, 8.83 mmol) in DCM (50 mL) at 0°C was added (3S)-hexahydropyridazine-3-carboxylic acid methyl ester; hydrochloride (compound B9, 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). After stirring 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 layers were 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 (10-30%) in petroleum ether to give (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Intermediate B, 2.4 g). MS calculated value 477 (MH + ), measured value 476.9 (MH + ).

[0260] Intermediate C

[0261] (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0262]

[0263] The title intermediate C was prepared according to the following scheme:

[0264]

[0265]

[0266] Step 1: Preparation of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (Compound C3)

[0267] To a mixture of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (Compound C1, 35.0 g, 116.8 mmol) in DCM (400 mL) was slowly added SnCl 4 Solution (97.2 mL, 121.5 mmol). After stirring at -40 °C for 0.5 h, a solution of 5-bromo-6-fluoro-1H-indole (Compound C2, 25.0 g, 116.8 mmol) in DCM (200 mL) was added dropwise to the mixture which was stirred at -40 °C for 15 min. After the reaction was completed, saturated NaHCO 3 The aqueous solution (800 mL) was quenched and the reaction mixture was extracted with EtOAc (900 mL, twice). The combined organic layers were washed with brine (700 mL) and purified by Na 2 SO 4 The mixture was dried, filtered and concentrated in vacuo. The residue was triturated with a solution (100 mL, petroleum ether: ethyl acetate = 8: 1) and filtered. The filter cake was dried in vacuo to give 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropane-1-one (Compound C3, 50.0 g) as a yellow solid. MS calculated value 552.1 (MH + ); measured value 552.1 (MH + ).

[0268] Step 2: Preparation of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C4)

[0269] To a mixture of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (Compound C3, 50.0 g, 90.49 mmol) in THF (600 mL) was added LiBH dropwise at 0°C. 4 (48.4 mL, 193.49 mmol, 4 M THF solution). The mixture was stirred at 70 ° C for 24 hours under a nitrogen atmosphere. After the reaction was completed, it was quenched by slowly adding water (600 mL) at 0 ° C, and the reaction mixture was extracted with EtOAc (600 mL, twice). The combined organic layer was washed with brine (600 mL) and purified by Na 2 SO 4 Dry, filter and concentrate in vacuo. The residue is purified by silica gel column chromatography Purification gave [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C4, 46.0 g) as a white solid. MS calculated value 538.1 (MH + ); measured value 538.2 (MH + ).

[0270] Step 3: Preparation of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5)

[0271] To a mixture of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C4, 35.4 g, 65.73 mmol) and iodine (18.4 g, 72.3 mmol) in THF (400 mL) at 0°C was added silver trifluoromethanesulfonate (20.3 g, 78.88 mmol). The mixture was stirred at 0°C for 10 min. After the reaction was complete, saturated Na 2 SO 3 The reaction was quenched with aqueous solution (400 mL) and EtOAc (400 mL) and the reaction mixture was filtered. The organic layer was washed with brine (100 mL) and purified by Na 2 SO 4 Dry, filter and concentrate in vacuo. The residue is purified by silica gel column chromatography Purification gave [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 43.0 g) as a yellow solid. MS calculated 664.0 (MH + ); measured value 664.1 (MH + ).

[0272] Step 4: Preparation of 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C6)

[0273] To a mixture of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 16.7 g, 25.13 mmol) and 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (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)Cl 2 (920mg, 1.26mmol). The mixture was stirred at 70 ° C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the mixture was filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 20% to 50%) to give 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C6, 19.5g) as a white solid. MS calculated value 891.3 (MH + ); measured value 891.3 (MH + ).

[0274] Step 5: Preparation of 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C7)

[0275] At 0°C, 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylate (Compound C6, 14.5 g, 16.26 mmol) and Cs 2 CO 3 (15.9 g, 48.77 mmol) was added dropwise to a solution of trifluoromethanesulfonic acid 2,2,2-trifluoroethyl ester (37.7 g, 162.56 mmol) in DMF (200 mL), and the mixture was stirred at 20 ° C for 12 hours. 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). The combined organic layer was washed with brine (100 mL, four times) and washed with Na2 SO 4 The mixture was dried, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica gel column chromatography to give 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C7, 8.0 g, PEAK 1, faster elution) as a yellow oil. MS calculated value 973.3 (MH + ); measured value 973.2 (MH + ).

[0276] Step 6: Preparation of 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C8)

[0277] To a solution of 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C7, 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 hours. After the reaction was complete, 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 layers were washed with brine (80 mL, three times) and purified by Na 2 SO 4 Dry, filter, and concentrate under vacuum to give a residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 25% to 66%) to give 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C8, 6.5 g) as a yellow solid. MS calculated value 735.2 (MH + ); measured value 735.1 (MH + ).

[0278] Step 7: Preparation of 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C9)

[0279] To a solution of benzyl 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylate (compound C8, 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)Cl 2 (537.1mg, 0.73mmol). The mixture was degassed and purged with a nitrogen atmosphere three times, and the mixture was stirred at 90 ° C for 12 hours. 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 obtain a residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 25% to 66%) to obtain 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1-(2,2,2-trifluoroethyl) indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl] piperazine-1-carboxylic acid benzyl ester (Compound C9, 5.2g) as a yellow oil. MS calculated value 783.3 (MH + ); measured value 783.3 (MH + ).

[0280] Step 8: Preparation of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound C10)

[0281] To a mixture of (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Intermediate B, 2.7 g, 5.69 mmol), 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (Compound C9, 4.9 g, 6.32 mmol) in toluene (60 mL) / 1,4-dioxane (20 mL) / water (20 mL) was added K under nitrogen atmosphere. 3 PO 4 (3.4g, 15.81mmol) and Pd(dtbpf)Cl 2 (412.2 mg, 0.63 mmol). The mixture was stirred at 70 ° C for 12 hours. After the reaction was completed, the mixture was concentrated in vacuo to give a residue. The residue was purified by silica gel column (EtOAc in PE = 10% to 75%) to give (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazine-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound C10, 3.6 g) as a brown solid. MS calculated value 1053.4 (MH + ); measured value 1053.3 (MH + ).

[0282] Step 9: Preparation of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid (Compound C11)

[0283] To a solution of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propionyl]-hexahydropyridazine-3-carboxylic acid methyl ester (Compound C10, 3.6 g, 3.42 mmol) in DCE (50 mL) was added trimethyltin alcohol (2.4 g, 13.67 mmol) and the mixture was stirred at 60 ° C for 12 hours. After the reaction was complete, 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 layers were washed with brine (100 mL) and purified by Na 2 SO 4 The mixture was dried, filtered and concentrated under vacuum to give (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid (Compound C11, 4.3 g) as a brown solid. MS calculated value 1039.4 (MH + ); measured value 1039.2 (MH + ).

[0284] Step 10: 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9 ,13 .0 22,26 Preparation of 1-(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl)-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C12)

[0285] To a mixture of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid (Compound C11, 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 hours. After the reaction was complete, the mixture was concentrated in vacuo, then diluted with water (80 mL) and extracted with EtOAc (80 mL, twice). The combined organic layers were washed with brine (80 mL) and purified by Na 2 SO 4 The residue was purified by silica gel column chromatography (EtOAc in PE = 25% to 66%) to give 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-tetraazapentacyclo[17.5.2.1]] as a yellow gum. 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C12, 3.1 g). MS calculated value 1021.4 (MH + ); measured value 1021.2 (MH + ).

[0286] Step 11: N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of tert-butyl 1(25),2,5(28),19,22(26),23-hexaen-7-yl)carbamate (Compound C13)

[0287] To 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22 ,26 To a mixture of octacosyl-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (Compound C12, 3.1 g, 3.04 mmol) and aqueous formaldehyde solution (775.0 mg, 9.55 mmol) in methanol (150 mL) was added Pd(OH) on activated carbon 2 (2.79 g, 3.97 mmol). The mixture was degassed and washed with H 2 The mixture was purged three times. The mixture was hydrogenated at 30°C for 18 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated in vacuo to give N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a brown solid. 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamic acid tert-butyl ester (Compound C13, 2.6 g). MS calculated value 901.3 (MH + ); measured value 901.3 (MH + ).

[0288] Step 12: (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of octadecane-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C)

[0289] To N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 To a mixture of tert-butyl]octadecanoate-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (compound C13, 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 complete, the mixture was concentrated in vacuo and washed with saturated NaHCO 3 (30 mL) and extracted with EtOAc (30 mL, three times). The combined organic layers were washed with brine (50 mL) and purified by Na 2 SO 4 The mixture was dried, filtered and concentrated in vacuo to give (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow solid. 2,5 .1 9,13 .0 22 ,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C, 2.0 g), which was used directly in the next step. MS: calculated value 801.3 (MH + ); measured value 801.2 (MH + )

[0290] Intermediate D

[0291] (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0292]

[0293] The title compound was prepared analogously to the preparation of intermediate C by using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate.

[0294] Intermediate E

[0295] (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0296]

[0297] The compounds were prepared according to the following scheme:

[0298]

[0299]

[0300] Step 1: Preparation of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2).

[0301] To a mixture of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3, 2.03 g, 5.95 mmol) and 1-(2,2,2-trifluoroethyl)piperazine (Compound E1, 1.0 g, 5.95 mmol) in toluene (15 mL) was added Cs 2 CO 3 (4.85 g, 14.88 mmol), (R)-binap (92.6 mg, 0.15 mmol), and Pd(OAc) 2(66.8 mg, 0.3 mmol). The reaction mixture was degassed and purged with nitrogen 3 times, and the mixture was stirred at 100 ° C under a nitrogen atmosphere for 12 hours. After cooling 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 1- [5-bromo-6- [(1S) -1-methoxyethyl] -3-pyridinyl] -4- (2,2,2-trifluoroethyl) piperazine (compound E2, 2.0 g) as a yellow oil. MS: calculated value 382.2 (MH + ); measured value 382.1 (MH + )

[0302] Step 2: 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3).

[0303] To a solution of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2, 3.2 g, 8.37 mmol), bis(pinacolato)diboron (3.19 g, 12.56 mmol) and KOAc (2.1 g, 20.93 mmol) in toluene (50 mL) was added Pd(dppf)Cl 2 (306.3 mg, 0.42 mmol). The mixture was degassed and purged with nitrogen 3 times, and the mixture was stirred at 90 ° C under a nitrogen atmosphere for 12 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue, which was purified by a reverse phase column to give 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3, 1.9 g) as a yellow gum. MS calculated value 430.2 (MH + ); measured value 348.4 (MC 6 H 10 +H + ).

[0304] Step 3: Preparation of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E4).

[0305] To a solution of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3, 1.9 g, 4.41 mmol), [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 2.1 g, 3.15 mmol) in 1,4-dioxane (24 mL), water (8 mL) and toluene (8 mL) was added K 3 PO 4 (2.1 g, 9.5 mmol) and Pd(dppf)Cl 2 (231 mg, 0.37 mmol). The mixture was degassed by bubbling nitrogen for 2 min, and the reaction mixture was stirred at 70 ° C for 12 hours. After cooling to room temperature, the reaction mixture was filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE: 30%-60%) to give [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazine-1-yl]-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E4, 960.0 mg) as a yellow gum. MS: Calculated value 839.3 (MH + ); measured value 839.3 (MH + )

[0306] Step 4: Preparation of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E5).

[0307] To a solution of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E4, 1 g, 1.14 mmol) in DMF (35 mL) was added Cs 2 CO 3(1.1 g, 3.44 mmol) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (2.7 g, 11.63 mmol). After stirring at 20 °C for 15 hours, the reaction mixture was poured into water (100 mL) and extracted with EtOAc (50 mL, three times). The combined organic phase was washed with brine (50 mL, three times) and purified by Na 2 SO 4 Dry, filter and concentrate under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE: 30%-40%) to give [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E5, 640.0 mg, faster elution) as a white solid. MS calculated value 921.3 (MH + ); measured value 921.4 (MH + ).

[0308] Step 5: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E6).

[0309] To a solution of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E5, 640.0 mg, 0.69 mmol) in DMF (7 mL) was added cesium fluoride (421.8 mg, 2.78 mmol). The mixture was stirred at 60 ° C for 16 hours. After cooling 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 (EtOAc in PE: 30%-60%) to give 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E6, 360.0 mg) as a yellow oil. MS calculated 683.2 (MH + ); measured value 683.1 (MH+ ).

[0310] Step 6: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E7).

[0311] To a solution of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E6, 360.0 mg, 0.53 mmol), bis(pinacolato)diboron (200.6 mg, 0.79 mmol) in toluene (6 mL) were added potassium acetate (0.08 mL, 1.32 mmol) and Pd(dppf)Cl 2 (40mg, 0.1mmol). The reaction mixture was degassed by bubbling nitrogen for 5min and then stirred at 80°C for 15 hours. After cooling 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 (EtOAc in PE: 30%-50%) to give 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazine-1-yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propane-1-ol (Compound E7, 300.0mg) as a yellow gum. MS calculated value 731.4 (MH + ); measured value 731.4 (MH + ).

[0312] Step 7: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound E8).

[0313] To a mixture of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E7, 0.3 g, 0.41 mmol) and (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Intermediate B, 196.7 mg, 0.41 mmol) in toluene (3 mL), 1,4-dioxane (1 mL) and water (1 mL) was added K 3 PO 4 (221.3 mg, 1.04 mmol) and Pd(dtbpf)Cl 2 (27.05mg, 0.04mmol). The mixture was stirred at 70 ° C for 12 hours under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (EtOAc in PE: 60%-80%) to obtain (3S) -1- [(2S) -2- (tert-butoxycarbonylamino) -3- [4- [6- fluoro-3- (3-hydroxy-2,2-dimethyl-propyl) - (2M) -2- [2- [(1S) -1- methoxyethyl] -5- [4- (2,2,2- trifluoroethyl) piperazine -1- base] -3- pyridyl] -1- (2,2,2- trifluoroethyl) indol -5- base] thiazol-2- base] propionyl] hexahydropyridazine -3- carboxylic acid methyl ester (compound E8, 200.0mg). MS calculated value 1001.4 (MH + ); measured value 1001.4 (MH + ).

[0314] Step 8: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound E9).

[0315] To a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound E8, 200.0 mg, 0.2 mmol) in DCE (5 mL) was added MeOH. 3 SnOH (200.0 mg, 1.11 mmol). The mixture was stirred at 60 °C for 12 hours. The reaction mixture was concentrated under vacuum to give a residue. EtOAc (10 mL) and water (10 mL) were added to the residue and the layers were separated. The aqueous phase was extracted with EtOAc (15 mL, twice). The combined organic layers were washed with brine (20 mL) and purified by Na 2 SO 4 The mixture was dried, filtered, and concentrated in vacuo to give (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound E9, 188.0 mg) as a brown solid. MS calculated value 987.4 (MH + ); measured value 987.4 (MH + ).

[0316] Step 9: N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of tert-butyl]octacosahedral-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound E10).

[0317] To a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound E9, 188.0 mg, 0.19 mmol) in DCM (20 mL) at 0°C were added DIEA (0.7 mL, 3.81 mmol), EDCI (550.0 mg, 2.87 mmol) and HOBt (65.0 mg, 0.48 mmol). After stirring at 20°C for 12 hours, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (20 mL, three times). The combined organic layers were washed with brine (30 mL) and purified by Na 2 SO 4 The mixture was dried, filtered and concentrated under vacuum to give a residue which was purified by column chromatography (EtOAc in PE: 50%-70%) to give N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow solid. 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamic acid tert-butyl ester (Compound E10, 110.0 mg). MS calculated value 969.4 (MH + ); measured value 969.5 (MH + ).

[0318] Step 10: (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]- Preparation of octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E).

[0319] To N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 To a solution of tert-butyl]octadecanoyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (compound E10, 110.0 mg, 0.11 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 complete, the reaction mixture was concentrated under vacuum to give a residue. Saturated NaHCO 3 The mixture was added with 4-nitro-2-nitropropene (20 mL) and the mixture was extracted with EtOAc (15 mL, twice). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford (7S, 13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow solid. 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E, 98.0 mg). MS calculated value 869.4 (MH + ); measured value 869.2 (MH + ).

[0320] Intermediate F

[0321] (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0322]

[0323] The title compound was prepared analogously to the preparation of intermediate E by using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate.

[0324] Intermediate G

[0325] (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9 ,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0326]

[0327] The title compound was prepared analogously to the preparation of intermediate E by using iodoethane and morpholine instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate and 1-(2,2,2-trifluoroethyl)piperazine (Compound E1).

[0328] Intermediate H

[0329] (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0330]

[0331] The title compound was prepared in analogy to the preparation of intermediate C by using 5-bromo-4-fluoro-1H-indole instead of 5-bromo-6-fluoro-1H-indole (compound C2).

[0332] Intermediate I

[0333] (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0334]

[0335] The title compound was prepared in analogy to the preparation of intermediate E by using morpholine instead of 1-(2,2,2-trifluoroethyl)piperazine (compound E1).

[0336] Example 1

[0337] (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide

[0338]

[0339] To a solution of (1S,2S)-2-methylcyclopropanecarboxylic acid (Compound 1a, 64.3 mg, 0.64 mmol) in DMF (2 mL) at 0 °C were added DIEA (0.3 mL, 1.61 mmol), HATU (366.5 mg, 0.96 mmol) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[175.2.1 2,5 .1 9,13 .0 22,26] octaconic acid-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D, 240.0 mg, 0.32 mmol). After stirring at 20°C for 16 hours, the reaction mixture was poured into ice water (10 mL) and extracted with EA (20 mL, three times). The combined organic layers were washed with brine (20 mL) and purified by Na 2 SO 4 The resulting residue was purified by silica gel chromatography to afford (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a white solid. 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide (Example 1, 156.41 mg). MS calculated value 829.5 (MH + ); measured value 829.5 (MH + ). 1 H NMR (400 MHz, methanol-d 4)δ=8.64(d,J=7.6Hz,1H),8.58(d,J=8.4Hz,1H),8.42(d,J=2.9Hz,1H),7.63(d,J=2 .4Hz,1H),7.34(d,J=2.8Hz,1H),7.31(d,J=12.7Hz,1H),5.79-5.73(m,1H),4.44(br d,J=11.4Hz,1H),4.28-4.13(m,4H),3.77-3.68(m,2H),3.41-3.37(m,4H),3.28-3.13(m,2H),3.02(br d,J=13.6Hz,1H),2.77-2.74(m,4H),2.63(br d,J=14.3Hz,1H),2.44(s,3H),2.21-2.13(m,1H),1.97(s,2H),1.68-1.58(m,1H),1.51- 1.47(m,1H),1.42(d,J=6.2Hz,2H),1.37(d,J=6.6Hz,4H),1.13(d,J=6.0Hz,3H),1.08(br dd,J=4.2,8.6Hz,1H),0.98(t,J=7.0Hz,3H),0.95-0.89(m,4H),0.67-0.62(m,1H),0.50(s,3H)ppm.

[0340] Example 2

[0341] (1R,5S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-oxabicyclo[3.1.0]hexane-6-carboxamide

[0342]

[0343] The title compound was prepared by using (1S,5R)-3-oxabicyclo[3.1.0]hexane-6-carboxylic acid instead of (1S,2S)-2-methylcyclopropanecarboxylic acid (Compound 1a) in analogy to the preparation of Example 1. Example 2 (10.2 mg) was obtained as a white solid. MS calculated value 857.4 (MH + ); measured value 857.1 (MH+ ). 1 H NMR (400 MHz, acetonitrile-d 3 )δ=8.61(d,J=7.6Hz,1H),8.41(d,J=2.8Hz,1H),7.62(d,J=2.5Hz,1H),7.33-7.28(m,1H),7.18(d,J=2.8Hz,1H),7.04(br d,J=9.5Hz,1H),5.68(br t,J=8.8Hz,1H),4.41-4.30(m,2H),4.20-4.05(m,4H),3.88-3.83(m,2H),3.71-3.64(m,4H),3.37(br d,J=14.9Hz,1H),3.28-3.24(m,4H),3.22-3.17(m,3H),3.13(br dd,J=9.1,15.0Hz,1H),2.91(br d,J=14.9Hz,1H),2.70(dt,J=2.7,12.9Hz,1H),2.58(br d,J=14.1Hz,1H),2.50(br t,J=4.8Hz,4H),2.30-2.22(m,4H),1.78-1.72(m,1H),1.57-1.50(m,2H),1.36(d,J=6.1Hz,3H),1.27(brs,1H),0.96(br t,J=7.1Hz,3H),0.88(s,3H),0.47(s,3H)ppm.

[0344] Example 3

[0345] (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide

[0346]

[0347] The title compound was prepared similarly to the preparation of Example 1 by using (1S,2S)-2-(difluoromethyl)cyclopropanecarboxylic acid instead of (1S,2S)-2-methylcyclopropanecarboxylic acid (Compound 1a). Example 3 (31.4 mg) was obtained as a yellow solid. MS calculated value 865.4 (MH + ); measured value 865.1 (MH + ). 1 H NMR (400MHz, chloroform-d) δ = 8.76 (br d, J = 2.6Hz, 1H), 8.66 (d, J = 7.6Hz, 1H), 7.63 (d, J = 2.1Hz, 1H), 7.30 (br d, J = 2.4Hz, 1H), 7.12 (d, J = 12.3Hz, 1H), 6.78 (br d,J=9.9Hz,1H),6.00-5.92(m,1H),4.66-4.53(m,1H),4.36(q,J=6.2Hz,1H),4.31-4.23(m,1H),4.19 -4.04(m,2H),3.89-3.86(m,1H),3.78-3.66(m,4H),3.47-3.44(m,1H),3.41(s,3H),3.24-3.05(m,6H ),2.93(s,3H),2.89-2.80(m,1H),2.78-2.70(m,1H),2.53-2.39(m,1H),2.29-2.18(m,1H),2.08-1.6 6(m,5H),1.53-1.45(m,3H),1.19-1.14(m,2H),1.37-1.08(m,2H),1.02-0.87(m,6H),0.50(s,3H)ppm.

[0348] Example 4

[0349] (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide

[0350]

[0351] Similar to the preparation of Example 1, by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C) was substituted with (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) to prepare the title compound. Example 4 (15.7 mg) was obtained as a white solid. MS calculated value 883.5 (MH + ); measured value 883.1 (MH + ). 1 H NMR (400 MHz, methanol-d 4 )δ=8.67(d,J=7.6Hz,1H),8.50(d,J=2.8Hz,1H),7.70(d,J=2.0Hz,1H),7.54-7.49(m,1H),7.49-7.45(m,1H),5.7 1(d,J=9.2Hz,1H),5.20-5.14(m,1H),4.47-4.41(m,1H),4.24-4.18(m,2H),4.09-3.94(m,2H),3.80-3.61(,5H), 3.49-3.43(m,2H),3.35(s,3H),3.17-3.12(m,2H),3.00(s,3H),2.83-2.76(m,1H),2.57(d,J=14.4Hz,1H),2.23- 2.16(m,1H),1.99-1.93(m,1H),1.84-1.77(m,1H),1.68-1.61(m,1H),1.52-1.48(m,1H),1.47-1.41(m,4H),1.27

[0352] -1.22(m,1H),1.13(d,J=6.0Hz,3H),1.11-1.05(m,2H),0.96(s,3H),0.90-0.77(m,1H),0.68-0.62(m,1H),0.44(s,3H).

[0353] Example 5 and Example 6

[0354] (1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide and (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide

[0355]

[0356] The compounds were prepared according to the following scheme:

[0357]

[0358] Step 1: (1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide and (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of 1-(25),2,5(28),19,22(26),23-hexaen-7-yl)cyclopropanecarboxamide

[0359] To a solution of trans-2-cyanocyclopropanecarboxylic acid (compound 5a, 11.2 mg, 0.1 mmol) in DMF (0.5 mL) was added DIEA (0.1 mL, 0.33 mmol), HATU (76.4 mg, 0.2 mmol) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[175.2.1] at 0 °C. 2,5 .1 9,13 .0 22,26 ] 28-hexadecane-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D, 50.0 mg, 0.07 mmol). After stirring at 20°C for 16 hours, the reaction mixture was prepared by preparative HPLC and SFC to give Example 5 (13.4 mg, faster elution) and Example 6 (9.9 mg, slower elution) as a white solid.

[0360] Example 5: MS calculated value 840.4 (MH + ); measured value 840.1 (MH + ). 1H NMR (400MHz, chloroform-d) δ = 8.63 (d, J = 7.3Hz, 1H), 8.49 (d, J = 2.8Hz, 1H), 7.59 (d, J = 2.0Hz, 1H), 7.12-7.07 (m, 2H), 6.88 (br d,J=8.4Hz,1H),5.92(t,J=8.9Hz,1H),4.60(br dd,J=2.4,12.5Hz,1H),4.30-4.19(m,3H),4.08-3.99(m,2H),3.85(br d,J=11.1Hz,1H),3.74-3.70(m,1H),3.49(br d,J=3.4Hz,4H),3.36(s,3H),3.20-3.13(m,2H),2.76-2.68(m,4H),2.63(s,1H),2.46(br d,J=14.2Hz,1H),2.25-2.20(m,2H),2.01(s,1H),1.95-1.90(m,2H),1.58(td,J=5.0,9.8Hz,2 H),1.47-1.42(m,5H),1.30-1.22(m,4H),0.97(t,J=7.1Hz,3H),0.91(s,2H),0.47(s,3H)ppm.

[0361] Example 6: MS calculated value 840.4 (MH + ); measured value 840.1 (MH + ). 1H NMR (400MHz, chloroform-d) δ = 8.64 (d, J = 7.6Hz, 1H), 8.51 (d, J = 2.9Hz, 1H), 7.63 (d, J = 2.2Hz, 1H), 7.13-7.07 (m, 2H), 6.88 (d, J = 9.5Hz, 1H), 5. 95(t,J=8.9Hz,1H),4.65-4.57(m,1H),4.32-4.21(m,3H),4.12-3.96(m,2H),3.86(d,J=10.8Hz,1H),3.75(d,J=11.1Hz,1H),3.48(br d,J=14.7Hz,1H),3.37(s,3H),3.34-3.29(m,4H),3.19(dd,J=8.8,15.0Hz,1H),3.05(br d,J=14.2Hz,1H),2.76-2.62(m,5H),2.52(br d,J=14.3Hz,1H),2.41(s,3H),2.27-2.19(m,2H),2.04-2.00(m,2H),1.48-1.43 (m,4H),1.32-1.23(m,2H),1.00(t,J=7.1Hz,3H),0.92(s,4H),0.49(s,3H)ppm.

[0362] Example 7

[0363] (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide

[0364]

[0365] Similar to the preparation of Example 1, by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate F) was substituted with (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) to prepare the title compound. Example 7 (12.5 mg) was obtained as a yellow solid. MS calculated value 897.4 (MH + ); measured value 897.6 (MH + ). 1 H NMR (400 MHz, methanol-d 4 )δ=8.68(d,J=7.6Hz,1H),8.38(d,J=2.8Hz,1H),7.78(d,J=2.4Hz,1H),7.6 5(d,J=2.4Hz,1H),7.35(d,J=12.8Hz,1H),5.81(d,J=8.8Hz,1H),4.48-4.40 (m,1H),4.38-4.31(m,1H),4.30-4.19(m,1H),4.16-4.01(m,2H),3.81-3.6 8(m,2H),3.49-3.44(m,4H),3.43-3.40(m,1H),3.37(s,3H),3.21-3.12(m,3 H),3.04-3.95(m,1H),2.92-2.86(m,4H),2.81-2.70(m,2H),2.21-2.11(m, 1H),1.99-1.89(m,1H),1.84-1.70(m,1H),1.67-1.57(m,1H),1.51-1.46(m, 1H),1.44(d,J=6.4Hz,3H),1.25-1.19(m,1H),1.15-1.10(m,3H),1.09-1.05 (m,1H),1.04-1.00(m,3H),0.94(s,3H),0.66-0.61(m,1H),0.59(s,3H)ppm.

[0366] Example 8

[0367] (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide

[0368]

[0369] Similar to the preparation of Example 1, by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was substituted with (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) to prepare the title compound. Example 8 (135.4 mg) was obtained as a light yellow solid. MS calculated value 816.4 (MH + ); measured value 816.1 (MH + ). 1 H NMR (400 MHz, methanol-d 4)δ=8.70(d,J=7.5Hz,1H),8.37(d,J=2.7Hz,1H),7.95(br s,1H),7.65(d,J=2.3Hz,1H),7.37(d,J=12.6Hz,1H),5.82(br d,J=8.9Hz,1H),4.47-4.38(m,2H),4.31-4.23(m,1H),4.15-4.10(m,1H),4.05-3.98(m,1H),3.88(br t,J=4.7Hz,4H),3.79(br d,J=11.0Hz,1H),3.71(d,J=10.6Hz,1H),3.47-3.38(m,10H),3.00(br d,J=15.0Hz,1H),2.84-2.72(m,2H),2.19-2.12(m,1H),1.97-1.90(m,1H),1.64(br d,J=2.7Hz,1H),1.50-1.45(m,4H),1.23(br dd,J=4.2,9.5Hz,2H),1.12(d,J=5.9Hz,3H),1.05(q,J=6.8Hz,4H),0.96(s,3H),0.62(s,3H)ppm.

[0370] Example 9

[0371] (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide

[0372]

[0373] Similar to the preparation of Example 1, by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E) was substituted with (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) to prepare the title compound. Example 9 (27.7 mg) was obtained as an off-white solid. MS calculated value 951.4 (MH + ); measured value 951.2 (MH + ). 1 H NMR (400 MHz, methanol-d 4 )δ=8.69(d,J=7.6Hz,1H),8.42(d,J=2.8Hz,1H),7.69(d,J=2.4Hz,1H),7.56-7.43(m,2H),5.73(d,J=8.8Hz,1H),5.24-5.08(m,1H),4.83-4. 78(m,1H),4.49-4.39(m,1H),4.26-4.15(m,2H),3.82-3.65(m,2H),3. 48-3.38(m,5H),3.36(s,3H),3.30-3.25(m,1H),3.20-3.09(m,3H),2. 88(t,J=4.8Hz,4H),2.84-2.75(m,1H),2.67-2.59(m,1H),2.26-2.15( m,1H),1.99-1.91(m,1H),1.87-1.74(m,1H),1.70-1.56(m,1H),1.52- 1.47(m,1H),1.45(d,J=6.0Hz,3H),1.28-1.19(m,1H),1.15-1.10(m,3 H),1.10-1.05(m,1H),0.97(s,3H),0.71-0.59(m,1H),0.49(s,3H)ppm.

[0374] Example 10

[0375] (1S,2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide

[0376]

[0377] Similar to the preparation of Example 1, by using (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate H) was substituted with (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) to prepare the title compound. Example 10 (11.2 mg) was obtained as a white solid. MS calculated value 883.4 (MH + ); measured value 883.5 (MH + ). 1 H NMR (400 MHz, methanol-d 4)δ=8.50(d,J=2.8Hz,1H),7.52-7.37(m,4H),6.03-5.88(m,1H),5.19-5.07(m,1H),4.80-4.76(m,2H ),4.44-4.34(m,2H),4.12-3.96(m,3H),3.72-3.47(m,6H),3.41-3.33(m,2H),3.28-3.25(m,2H),3.1 9-3.05(m,5H),2.99(s,4H),2.59-2.52(m,1H),1.66-1.53(m,1H),1.44(d,J=6.0Hz,3H),1.39-1.29 (m,2H),1.26-1.17(m,2H),1.10-1.05(m,3H),1.05-1.01(m,1H),0.80(s,3H),0.65-0.52(m,3H)ppm.

[0378] Example 11

[0379] (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide

[0380]

[0381] Similar to the preparation of Example 1, by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) and (1S,2S)-2-(difluoromethyl)cyclopropanecarboxylic acid were used instead of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octadecane-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) and (1S,2S)-2-methylcyclopropanecarboxylic acid (Compound 1a) were used to prepare the title compound. Example 11 (33.2 mg) was obtained as a yellow solid. MS calculated value 852.4 (MH + ); measured value 852.2 (MH + ). 1H NMR (400MHz, chloroform-d) δ = 8.91 (br s, 1H), 8.66 (d, J = 7.6Hz, 1H), 7.62 (d, J = 2.2Hz, 1H), 7.41 (s, 1H), 7.13 (s, 1H), 7.10 (s, 1H), 6.75 (br d,J=8.9Hz,1H),5.99-5.93(m,1H),5.83(d,J=3.1Hz,1H),4.60(br d,J=11.9Hz,1H),4.38(br d,J=6.4Hz,1H),4.21(br d,J=9.2Hz,1H),4.13-4.06(m,2H),3.87(br d,J=11.4Hz,2H),3.79-3.65(m,2H),3.45(br d,J=15.2Hz,1H),3.41(s,2H),3.39-3.35(m,4H),3.17(brdd,J=8.6,15.0Hz,2H),3.10(br d,J=11.6Hz,1H),2.80-2.63(m,2H),2.50(br d,J=13.9Hz,1H),2.20(br d,J=10.9Hz,1H),1.99(br d,J=17.9Hz,1H),1.87-1.78(m,2H),1.67-1.59(m,1H),1.53(d,J=6.4Hz,3H),1.33-1 .21(m,3H),1.18-1.12(m,1H),1.01(brt,J=7.0Hz,3H),0.94(s,3H),0.53(s,3H)ppm.

[0382] Example 12

[0383] (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide

[0384]

[0385] Similar to the preparation of Example 1, by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate I) was substituted with (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) to prepare the title compound. Example 12 (150 mg) was obtained as a white solid. MS calculated value 870.4 (MH + ); measured value 870.5 (MH + ). 1 H NMR (400 MHz, methanol-d 4)δ=8.65-8.63(d,J=7.6Hz,1H),8.41-8.40(d,J=2.8Hz,1H),7.70-7.65(d,J=2.4Hz,1 H),7.50-7.40(d,J=12.4Hz,1H),7.31-7.28(d,J=2.4Hz,1H),5.76-5.66(d,J=8.0Hz,1 H),5.18-5.05(m,1H),4.96-4.88(m,1H),4.47-4.39(m,1H),4.23-4.10(m,2H),3.90- 3.83(t,J=4.8Hz,J=9.6Hz,4H),3.80-3.75(d,J=10.8Hz,1H),3.71-3.63(m,1H),3.49- 3.42(m,1H),3.30-3.29(m,3H),3.28-3.21(m,4H),3.14-3.07(d,J=14.4Hz,1H),2.86 -2.77(m,1H),2.63-2.55(d,J=14.4Hz,1H),2.23-2.14(m,1H),1.98-1.89(m,1H),1.84 -1.72(m,1H),1.65-1.56(m,1H),1.52-1.46(m,1H),1.45-1.40(d,J=6.4Hz,3H),1.30- 1.19(m,2H),1.13-1.06(m,4H),0.95(s,3H),0.69-0.58(m,1H),0.48-0.39(s,3H)ppm.

[0386] Biological Examples

[0387] Compound A122 from WO2022060836 (page 70 of Table 1a) is cited as a reference compound of the present invention.

[0388]

[0389] (A122)

[0390] Example 13

[0391] Cell viability assay

[0392] The purpose of this cellular assay is to determine the effect of the tested 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 the endpoint using the Cell Counting Kit-8.

[0393] Cells were seeded in 96-well assay plates (Corning-3699) at 5,000 cells / well (NCI-H358), 2,000 cells / well (AGS), and 2,000 cells / well (SW620) and incubated overnight. On the day of the assay, diluted compounds were added at a final concentration of 0.5% DMSO. After 72 hours of incubation, one-tenth the volume of Cell Counting Kit 8 (Dnjindo-CK04) was added to each well. After 2 hours of incubation, the signal was read using EnVision (OD450 minus OD650). IC 50 Determined by fitting a 4-parameter sigmoidal concentration response model.

[0394] Table 1. Activity of Examples and Compounds of the Invention in KRAS Cell Viability Assay

[0395] Examples G12C IC50(μM) G12D IC50(μM) G12V IC50(μM) A122 <0.001 0.001 <0.001 Example 1 0.018 4.630 0.009 Example 2 0.011 0.059 0.005 Example 3 0.0116 0.197 0.008 Example 4 0.019 1.265 0.013 Example 5 0.023 0.089 0.012 Example 6 0.079 0.316 0.035 Example 7 0.074 0.389 0.027 Example 8 0.016 1.493 0.013 Example 9 0.224 0.704 0.040 Example 10 0.012 0.013 0.003 Example 11 0.005 0.111 0.007 Example 12 0.011 0.244 0.016

[0396] Example 14

[0397] Interaction assay of KRAS-BRAF and CYPA (500 nM)

[0398] In this example, TR-FRET was also used to measure compound or compound-CYPA-dependent disruption of the KRAS G12C-BRAF complex. This protocol was also used to measure the disruption of KRAS G12D or KRAS G12V binding to BRAF by the compounds of the invention, respectively. In a 25 mM HEPES PH = 7.4 (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, Thermo, 15630080), 0.002% Tween20, 0.1% BSA, 100 mM NaCl, 5 mM MgCl 2 , 10 μM GMPPNP (guanosine 5'-[β,γ-imino] triphosphate trisodium salt hydrate, Sigma, G0635), untagged CYPA, GMPPNP loaded with 6His-KRAS protein, and GST-BRAF RBDMixed in the wells of a 384-well assay plate at final concentrations of 50 nM, 6.25 nM, and 1 nM, respectively. The compounds were present in the wells of the plate in a 16-point 3-fold dilution series starting from a final concentration of 10 μM 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 final concentrations of 6.67 nM and 0.21 nM, respectively, and the plate was incubated for another 1.5 hours. The TR-FRET signal was read on a PHERstar FSX microplate reader (Ex 320 nm, Em 665 / 615 nm). Compounds that promote the destruction of the KRAS-BRAF complex were identified as compounds that cause a decrease in the TR-FRET ratio compared to the DMSO control wells.

[0399] Table 2. Activity of Examples and Compounds of the Invention in the KRAS-BRAF and CYPA (500 nM) Interaction Assay

[0400] Examples <![CDATA[G12C IC 50 (μM)]]> <![CDATA[G12D IC 50 (μM)]]> <![CDATA[G12V IC 50 (μM)]]> A122 0.013 0.124 0.025 Example 1 0.108 0.915 0.211 Example 2 0.134 0.957 0.174 Example 3 0.196 1.715 0.243 Example 4 0.036 0.544 0.095 Example 5 0.194 0.862 0.258 Example 6 0.154 0.701 0.273 Example 7 0.192 >10 0.697 Example 8 0.160 1.674 0.143 Example 9 0.090 0.660 0.171 Example 10 0.009 0.020 0.013

[0401] Example 15

[0402] pERK inhibition assay

[0403] The assay is designed to measure the ability of the tested compounds to inhibit ERK phosphorylation, KRAS G12C in NCI-H358 cells, KRAS G12D in AGS cells, and KRAS G12V downstream signaling in SW620. NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL-1739) cells, and SW620 (ATCC-CCL-227) cells were grown and maintained in RPMI-1640 medium (Thermo Fisher Scientific) containing 10% fetal bovine serum and 1% penicillin / streptomycin. The day before the addition of the compound, the cells were plated at a density of 30,000 cells / well, 20,000 cells / well, and 30,000 cells / well (for NCI-H358, AGS, and SW620) in a tissue culture treated 96-well plate (Corning-3699) and allowed to attach overnight. The diluted compounds were then added at a final concentration of 0.5% DMSO. After 4 hours of incubation, the culture medium was removed, 100 μL 4% formaldehyde was added, and the assay plate was incubated at room temperature for 20 minutes. The plate was then washed once with phosphate buffered saline (PBS) and permeabilized with 100 μL frozen methanol for 10 minutes. Nonspecific antibodies bound to the plate were blocked at room temperature for at least 1 hour using 50 μL 1X BSA blocking buffer (Thermo-37520, diluted 10 times with phosphate buffered saline (PBST)).

[0404] The amount of phosphor-ERK was determined using an antibody specific for phosphorylated ERK. The primary antibody (pERK, CST-4370, Cell Signaling Technology) was diluted 1:300 in blocking buffer, with 50 μL aliquoted to each well and incubated overnight at 4 ° C. The cells were washed five times with PBST for 5 minutes. The secondary antibody (HRP-linked anti-rabbit IgG, CST-7074, Cell Signaling Technology) was diluted 1:1000 in blocking buffer, and 50 μL was added to each well, and incubated at room temperature for 1-2 hours. The cells were washed 5 times with PBST for 5 minutes, 100 μL TMB ELISA substrate (abcam-ab171523) was added, and gently shaken for 20 minutes. 50 μL stop solution (abcam-ab171529) was added, and then the signal (OD450) was read with EnVision.

[0405] IC 50 Determined by fitting a 4-parameter sigmoid concentration response model.

[0406] Table 3. Activity of Examples and Compounds of the Invention in the KRAS pERK Inhibition Assay

[0407] Examples <![CDATA[G12C IC 50 (μM)]]> <![CDATA[G12D IC 50 (μM)]]> <![CDATA[G12V IC 50 (μM)]]> A122 0.002 0.001 <0.001 Example 1 0.011 0.034 0.004 Example 2 0.009 0.018 0.003 Example 3 0.011 0.029 0.005 Example 4 0.02 0.047 0.007 Example 5 0.057 0.056 0.013 Example 6 0.079 0.116 0.025 Example 7 0.071 0.115 0.02 Example 8 0.023 0.056 0.008 Example 9 0.047 0.161 0.022 Example 10 0.017 0.011 0.002 Example 11 0.021 0.028 0.007 Example 12 0.033 0.053 0.009

Claims

1. A compound of formula (I), in R 1 is 3-oxabicyclo[3.1.0]hexane, C 1-6 Alkyl C 3-7 Cycloalkyl, cyano C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl; R 2 is H or halogen; R 3 is H or halogen; R 4 C 1-6 Alkyl or halogenated C 1-6 alkyl; R 5 C 1-6 Alkoxy C 1-6 alkyl; R 6 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl; A 1 is thiazolylidene; A 2 C 1-6 Alkylene; The condition is R 2 and R 3 Not at the same time H; or a pharmaceutically acceptable salt thereof.

2. A compound of formula (Ia), in R 1 is 3-oxabicyclo[3.1.0]hexane, C 1-6 Alkyl C 3-7 Cycloalkyl, cyano C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl; R 2 is H or halogen; R 3 is H or halogen; R 4 C 1-6 Alkyl or halogenated C 1-6 alkyl; R 5 C 1-6 Alkoxy C 1-6 alkyl; R 6 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl; A 1 is thiazolylidene; A 2 C 1-6 Alkylene; The condition is R 2 and R 3 Not at the same time H; or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 1 or 2, wherein R 1 C 1-6 Alkyl C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl.

4. A compound according to any one of claims 1 to 3, wherein R 1 It is methylcyclopropyl or (difluoromethyl)cyclopropyl.

5. A compound according to any one of claims 1 to 4, wherein R 2 is H or fluorine.

6. A compound according to any one of claims 1 to 5, wherein R 3 is H or fluorine.

7. A compound according to any one of claims 1 to 6, wherein R 4 It is ethyl or 2,2,2-trifluoroethyl.

8. A compound according to any one of claims 1 to 7, wherein R 5 It is 1-methoxyethyl.

9. A compound according to any one of claims 1 to 8, wherein R 6 is morpholinyl or C 1-6 Alkylpiperazinyl.

10. A compound according to any one of claims 1 to 9, wherein R 6 It is morpholinyl or 4-methylpiperazin-1-yl.

11. A compound according to any one of claims 1 to 10, wherein A 1 for Where bond "a" is attached to the indole ring.

12. A compound according to any one of claims 1 to 11, wherein A 2 It is 2,2-propylene.

13. The compound according to claim 1 or 2, wherein R 1 C 1-6 Alkyl C 3-7 Cycloalkyl or halogenated C 1-6 Alkyl C 3-7 Cycloalkyl; R 2 is H or halogen; R 3 is H or halogen; R 4 C 1-6 Alkyl or halogenated C 1-6 alkyl; R 5 C 1-6 Alkoxy C 1-6 alkyl; R 6 is morpholinyl or C 1-6 Alkylpiperazinyl; A 1 for wherein bond "a" is attached to the indole ring; A 2 C 1-6 Alkylene; The condition is R 2 and R 3 Not at the same time H; or a pharmaceutically acceptable salt thereof.

14. The compound according to claim 13, wherein R 1 is 2-methylcyclopropyl or 2-(difluoromethyl)cyclopropyl; R 2 is H or fluorine; R 3 is H or fluorine; R 4 is ethyl or 2,2,2-trifluoroethyl; R 5 is (1S)-1-methoxyethyl; R 6 is morpholinyl or 4-methylpiperazin-1-yl; A 1 for wherein bond "a" is attached to the indole ring; A 2 is 2,2-propylene; The condition is R 2 and R 3 Not at the same time H; or a pharmaceutically acceptable salt thereof.

15. A compound selected from: (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; (1R,5S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-oxabicyclo[3.1.0]hexane-6-carboxamide; (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; (1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide; (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octadecanoic acid-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9, 13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxamide; and (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; or a pharmaceutically acceptable salt thereof.

16. A method for preparing a compound according to any one of claims 1 to 15, comprising any one of the following steps: a) making a compound of formula (II), With acid (III), A coupling reaction is carried out in the presence of a coupling reagent and a base to form the compound of formula (I); Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , A 1 and A 2 As defined in any one of claims 1 to 14; the coupling reagent is T3P, HATU, PyBOP or EDCI / HOBt; the base is TEA, DIEPA or DMAP.

17. A compound or a pharmaceutically acceptable salt according to any one of claims 1 to 15 for use as a therapeutically active substance.

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15 and a pharmaceutically acceptable excipient.

19. Use of a compound according to any one of claims 1 to 15 for treating a KRAS G12C protein-related disease.

20. Use of a compound according to any one of claims 1 to 15 for treating KRASG12C, G12D and G12V protein related diseases.

21. Use of a compound according to any one of claims 1 to 15 for inhibiting the interaction of RAS with downstream effectors, wherein the downstream effectors are RAF and PI3K.

22. Use of a compound according to any one of claims 1 to 15 for inhibiting propagated oncogenic MAPK and PI3K signalling.

23. Use of a compound according to any one of claims 1 to 15 for treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma ovarian cancer and endometrial cancer.

24. Use of a compound according to any one of claims 1 to 15 for treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

25. A compound or pharmaceutically acceptable salt according to any one of claims 1 to 15 for use in treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

26. Use of a compound according to any one of claims 1 to 15 for the preparation of a medicament for treating or preventing a cancer driven by a KRAS mutation, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

27. A method of treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer, the method comprising administering a therapeutically effective amount of a compound as defined in any one of claims 1 to 15.

28. A compound or pharmaceutically acceptable salt according to any one of claims 1 to 15, prepared according to the process of claim 16.

29. The invention as herein described.

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