HK40118889A - Macrocycle compounds useful as kras inhibitors - Google Patents

Description

Macrocyclic compounds that can be used as KRAS inhibitors

This invention relates to organic compounds that can be used for treatment and/or prevention in mammals, and particularly to inhibitory effects on KRAS mutants that can be used to treat cancer.

Technical Field

RAS is one of the most well-known proto-oncogenes. Approximately 30% of human cancers contain mutations in the three most prominent members (KRAS, HRAS, and NRAS), making them the most common drivers of cancer development. KRAS mutations are often associated with poor prognosis, particularly in colorectal, pancreatic, and lung cancers. As the most frequently mutated RAS subtype, KRAS has been extensively studied in recent 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)). It is noteworthy that KRAS wild-type amplification was also found in approximately 7% of all cancers with KRAS alterations (ovarian, esophageal and gastric, and uterine), ranking among the top alterations.

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

KRAS proteins switch between inactive and active forms 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 SOS1 (Seven Sons homolog 1) or GTPase activators (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 ), thereby recruiting the RAF kinase family from the cytoplasm to the cell membrane, where they dimerize and become active. Activated RAFs then undergo a series of phosphorylation reactions on their downstream mitogen-activated protein kinases (MEK) and extracellular signal-regulated kinases (ERK), and propagate growth signals. Among the RAF protein kinase family (three known isoforms ARAF, BRAF, and CRAF/RAF1), BRAF mutations are most frequent and it remains the most potent activator of MEK. Although individual RAS and RAF family members exhibit different binding preferences, all RAFs possess a conserved RBD for forward transmission of MAPK signaling, frequently used to characterize KRAS inhibition (e.g., the KRAS-BRAF ^RBD in this paper). For KRAS, mutations at positions 12, 13, 61, and 146 lead to conversion to the active KRAS form by weakening nucleotide hydrolysis or activating nucleotide exchange, thereby resulting in overactivation of the tumorigenetic MAPK pathway.

Despite its widely recognized importance in cancerous malignancies, persistent efforts have failed to develop approved therapies for KRAS-mutant cancers until recently, when the first selective drug, AMG510, was rapidly approved as second-line treatment for KRAS G12C-driven non-small cell lung cancer (NSCLC). However, after approximately six months of treatment, clinically acquired resistance to KRAS G12C inhibitors emerges dramatically as the disease progresses. All mutations converge to reactivate RAS-MAPK signaling, with secondary RAS mutations already observed at oncogenic hotspots (e.g., G12/G13/Q61) and within switch II pockets (e.g., H95, R68, and Y96); furthermore, novel agents remain lacking in over 85% of all KRAS-mutant or wild-type amplification-driven cancers. In conclusion, both the numerous escape mechanisms and diverse oncogenic alleles underscore the urgent medical need for additional KRAS therapies. Therefore, we have invented oral compounds that target and inhibit the KRAS allele for the treatment of KRAS mutation-driven cancers.

Summary of the Invention

This invention relates to novel compounds having formula (I),

in

R ¹ is 3-oxabicyclo[3.1.0]hexyl, _C1-6 alkyl _C3-7 cycloalkyl, cyano _C3-7 cycloalkyl, or halo- _C1-6 alkyl _C3-7 cycloalkyl;

^R2 is H or a halogen;

^R3 is H or a halogen;

^R4 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

R ⁵ is a _C1-6 alkoxy- _C1-6 alkyl group;

^R6 is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino;

^A1 is an imidazolyl group;

^A2 is a _C1-6 alkylene group;

The condition is that ^R2 and ^R3 are not both H at the same time;

Or its medicinal salt.

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

Compounds of formula (I) or (Ia) exhibit good KRAS inhibitory activity against G12C and G12V. In another embodiment, the compounds of the present invention exhibit excellent cancer cell inhibitory activity and human hepatocyte stability. Furthermore, compounds of formula (I) or (Ia) also exhibit good or improved cytotoxicity and solubility characteristics. In addition, the compounds of the present invention have favorable pharmacokinetic properties compared to reference compounds.

Detailed Implementation

definition

The term " _C1-6 alkyl" refers to a saturated, straight-chain or branched alkyl group containing 1 to 6, particularly 1 to 4, carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, etc. In particular, the " _C1-6 alkyl" group is methyl, ethyl, or n-propyl.

The term “C _1-6 alkoxy” means C _1-6 alkyl-O-.

The term "( _C1-6 )alkylene" refers to a straight-chain or branched saturated divalent hydrocarbon group with 1 to 6 carbon atoms, or a divalent branched saturated divalent hydrocarbon group with 3 to 6 carbon atoms. Examples of ( _C1-6 )alkylene groups include methylene, ethylene, propylene, 2-methylpropylene, butylene, 2-ethylbutylene, pentylene, and hexylene.

The terms “halogen” and “halogenated” are used interchangeably in this document to refer to fluorine, chlorine, bromine, or iodine.

The term "halogenated _C1-6 alkyl" refers to a _C1-6 alkyl group in which at least one hydrogen atom of the _C1-6 alkyl group has been substituted by the same or different halogen atoms, particularly fluorine atoms. Examples of halogenated alkyl groups include monofluoro-, difluoro-, or trifluoro-methyl, -ethyl, or -propyl, such as 3,3,3-trifluoropropyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, fluoromethyl, or trifluoromethyl.

The term "C _3-7 cycloalkyl" refers to a monovalent saturated monocyclic or bicyclic hydrocarbon group with 3 to 7 ring carbon atoms. Bicyclic means consisting of two saturated carbon rings having one or two common carbon atoms. Examples of monocyclic cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl. Examples of bicyclic cycloalkyl groups are bicyclic [1.1.0]butyl, bicyclic [2.2.1]heptyl, bicyclic [1.1.1]pentyl, or bicyclic [2.2.2]octyl.

The term "thiazolyl" refers to a divalent thiazolyl group.

The term "oxo" refers to a divalent oxygen atom =O.

The term "2,2-propylidene" indicates

The term "protecting group" refers to a group that selectively blocks a reaction site in a multifunctional compound so that a chemical reaction can selectively occur at another unprotected reaction site that is typically associated with it in synthetic chemistry. Protecting groups can be removed at appropriate points in time. Exemplary protecting groups are amino protecting groups, carboxyl protecting groups, or hydroxyl protecting groups.

Those skilled in the art will understand that the following structures of compounds of formula (I) and (I') are identical, especially for the chiral centers:

The term "medicinal salt" refers to a salt that is not biologically or otherwise undesirable. "Medicinal salt" includes both acid addition salts and base addition salts.

"Pharmaceutical acid addition salts" refer to pharmaceutical salts formed with inorganic and organic acids. The inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, and phosphoric acid. The organic acids can be selected from aliphatic, alicyclic, aromatic, arylaliphatic, 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, dihydroxynaphthalic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid.

The term "medicinal base addition salt" refers to those medicinal salts that form 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 medicinal organic non-toxic bases include primary amines, secondary amines, tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and salts from 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, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, and polyamine resins).

The term "pharmaceutical active metabolite" refers to a pharmacologically active product produced through the metabolism of a specific compound or its salt in the body. Once in the human body, most drugs are substrates for chemical reactions that can alter their physical properties and biological effects. These metabolic transformations, which typically affect the polarity of the compounds of this invention, alter the way drugs are distributed and excreted from the body. However, in some cases, drug metabolism is essential for therapeutic efficacy.

The term "therapeutic effective amount" refers to the amount of a compound or molecule of the present invention, when administered to a subject, (i) to treat or prevent a particular disease, condition, or disorder; (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder; or (iii) to prevent or delay the onset of one or more symptoms of a particular disease, condition, or disorder described herein. Therapeutic 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 physician or veterinarian, and other factors.

The term "pharmaceutical composition" refers to a mixture or solution containing a therapeutically effective amount of an active pharmaceutical ingredient and a pharmaceutical excipient to be administered together to a mammal (e.g., a human) in need of such treatment.

The terms “medicinal excipient,” “medicinal carrier,” and “therapeutic inert excipient” are used interchangeably and refer to any of the following pharmaceutical ingredients in a pharmaceutical composition that are not therapeutically active and are non-toxic to the subject to which they are administered, such as disintegrants, binders, fillers, solvents, buffers, tensioning agents, stabilizers, antioxidants, surfactants, carriers, diluents, or lubricants used in the formulation of pharmaceutical products.

KRAS inhibitors

This invention relates to (i) a compound of formula (I),

in

R ¹ is 3-oxabicyclo[3.1.0]hexyl, _C1-6 alkyl _C3-7 cycloalkyl, cyano _C3-7 cycloalkyl, or halo- _C1-6 alkyl _C3-7 cycloalkyl;

^R2 is H or a halogen;

^R3 is H or a halogen;

^R4 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

R ⁵ is a _C1-6 alkoxy- _C1-6 alkyl group;

^R6 is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino;

^A1 is an imidazolyl group;

^A2 is a _C1-6 alkylene group;

The condition is that ^R2 and ^R3 are not both H at the same time;

Or its medicinal salt.

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

in

R ¹ is 3-oxabicyclo[3.1.0]hexyl, _C1-6 alkyl _C3-7 cycloalkyl, cyano _C3-7 cycloalkyl, or halo- _C1-6 alkyl _C3-7 cycloalkyl;

^R2 is H or a halogen;

^R3 is H or a halogen;

^R4 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

R ⁵ is a _C1-6 alkoxy- _C1-6 alkyl group;

^R6 is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino;

^A1 is an imidazolyl group;

^A2 is a _C1-6 alkylene group;

The condition is that ^R2 and ^R3 are not both H at the same time;

Or its medicinal salt.

A further embodiment of the invention is (iii) a compound of formula (I) or (Ia) according to (i) or (ii), or a pharmaceutical salt thereof, wherein ^R1 is a _C1-6 alkyl- _C3-7 cycloalkyl or a halogenated _C1-6 alkyl- _C3-7 cycloalkyl.

A further embodiment of the invention is (iv) a compound of formula (I) or (Ia) according to any one of (i) to (iii) or a pharmaceutical salt thereof, wherein ^R1 is methylcyclopropyl or (difluoromethyl)cyclopropyl.

A further embodiment of the invention is (v) a compound of formula (I) or (Ia) according to any one of (i) to (iv), wherein ^R2 is H or fluorine.

A further embodiment of the invention is (vi) a compound of formula (I) or (Ia) according to any one of (i) to (v), or a pharmaceutical salt thereof, wherein ^R3 is H or fluorine.

A further embodiment of the invention is (vii') a compound of formula (I) or (Ia) according to any one of (i) to (vi), or a pharmaceutical salt thereof, wherein ^R4 is ethyl or 2,2,2-trifluoroethyl.

A further embodiment of the invention is (viii') a compound of formula (I) or (Ia) according to any one of (i) to (vii), or a pharmaceutical salt thereof, wherein ^R5 is 1-methoxyethyl.

A further embodiment of the invention is (ix) a compound of formula (I) or (Ia) according to any one of (i) to (viii), or a pharmaceutical salt thereof, wherein ^R6 is morpholino or _C1-6 alkylpiperazinyl.

A further embodiment of the invention is (x) a compound of formula (I) or (Ia) according to any one of (i) to (ix), or a pharmaceutical salt thereof, wherein ^R6 is morpholino or 4-methylpiperazin-1-yl.

A further embodiment of the invention is (xi) a compound of formula (I) or (Ia) according to any one of (i) to (x), or a pharmaceutical salt thereof, wherein ^A1 is a compound in which bond “a” is attached to an indole ring.

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 pharmaceutical salt thereof, wherein ^A2 is 2,2-propylidene.

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

^R1 is a _C1-6 alkyl- _C3-7 cycloalkyl or a halogenated _C1-6 alkyl- _C3-7 cycloalkyl;

^R2 is H or a halogen;

^R3 is H or a halogen;

^R4 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

R ⁵ is a _C1-6 alkoxy- _C1-6 alkyl group;

^R6 is morpholino or _C1-6 alkylpiperazino;

^A1 is where the "a" bond is attached to the indole ring;

^A2 is a _C1-6 alkylene group;

The condition is that ^R2 and ^R3 are not both H at the same time;

Or its medicinal salt.

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

^R1 is 2-methylcyclopropyl or 2-(difluoromethyl)cyclopropyl;

^R2 is H or fluorine;

^R3 is H or fluorine;

^R4 is ethyl or 2,2,2-trifluoroethyl;

R ⁵ is (1S)-1-methoxyethyl;

^R6 is morpholino or 4-methylpiperazin-1-yl;

^A1 is where the "a" bond is attached to the indole ring;

^A2 is 2,2-propylidene;

The condition is that ^R2 and ^R3 are not both H at the same time;

Or its medicinal salt.

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

(1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexene-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]cyclopropaneformamide;

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

(1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]cyclopropaneformamide;

(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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropanecarboxamide;

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

Or its medicinal salt.

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 steps of:

a) Make compound of formula (II),

It undergoes a coupling reaction with acid (III) in the presence of a coupling agent and a base to form compound (I);

^R1 , ^R2 , ^R3 , ^R4 , ^R5 , ^R6 , ^R7 , ^A1 and ^A2 are as defined in any one of (i) to (xiv); the coupling agent is _T3P , HATU, PyBOP or EDCI/HOBt; the base is TEA, DIEPA or DMAP.

Another embodiment of the present invention is (xvii) a compound or pharmaceutical salt according to any one of (i) to (xv) which is used as a therapeutically active substance.

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

Another embodiment of the invention is (xix) the use of the compound according to any one of (i) to (xv) for the treatment of KRAS G12C protein-related diseases.

Another embodiment of the present invention is (xx) the use of the compound according to any one of (i) to (xv) for the treatment of diseases related to KRAS G12C, G12D and G12V proteins.

Another embodiment of the invention is (xxi) the use of the compound according to any one of (i) to (xv) for suppressing the interaction between RAS and downstream effectors, wherein the downstream effectors are RAF and PI3K.

Another embodiment of the invention is (xxii) the use of the compound according to any one of (i) to (xv) for inhibiting the transmission of carcinogenic MAPK and PI3K signaling.

Another embodiment of the invention is (xxiii) the use of the compound according to any one of (i) to (xv) for the treatment or prevention of 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.

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

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

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

Another embodiment of the present invention is (xxvii) a method for treating or preventing 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).

Another embodiment of the present invention is (xxviii) a compound or pharmaceutical salt according to any one of (i) to (xv), which is manufactured according to the method of (xvi).

Pharmaceutical composition and administration

Another embodiment provides pharmaceutical compositions or medicaments containing the compounds of the present invention and a therapeutically inert carrier, diluent, or excipient, and methods for preparing such compositions and medicaments using the compounds of the present invention. In one example, the compound of formula (I) can be formulated into a galenical form by mixing it with a physiologically acceptable carrier (i.e., a carrier that is non-toxic to the recipient at the dose and concentration used) at ambient temperature, at a suitable pH, and with the 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.

The composition shall be formulated, measured, and administered in accordance with good medical practice. Factors to be considered in this context include the specific disease being treated, the specific mammal being treated, the individual patient's clinical condition, the cause of the disease, the site of delivery, the method of administration, the timing of administration, and other factors known to the practicing physician. The “effective amount” of the compound to be administered will be influenced by these considerations and is the minimum amount necessary to inhibit the interaction between mutant RAS (e.g., KRAS G12C) and RAF, thereby blocking oncogenic MAPK signaling. For example, this amount may be below what would be toxic to normal cells or the mammal as a whole.

In one example, the effective amount of the compound of the invention per dose administered parenterally will be in the range of about 0.1 to 1000 mg/kg of patient body weight per day, alternatively in the range of about 0.1 to 1000 mg/kg of patient body weight, with the initial range of the compound typically being 0.3 to 15 mg/kg/day. 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 invention.

The compounds of this invention can be administered by any suitable manner, including oral, topical (including buccal and sublingual), rectal, vaginal, percutaneous, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal, epidural, and intranasal administration, as well as (if necessary for local treatment) intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

The compounds of this invention can be administered in any convenient form, such as tablets, powders, capsules, solutions, dispersants, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional to pharmaceutical formulations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and other active agents.

Conventional formulations are prepared by mixing the compounds of the present invention with a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail, for example, in Ansel, Howard C. et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R. et al. , Remington: The Science and Practice of Pharmacy . Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients . Chicago, Pharmaceutical Press, 2005. The formulation may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow aids, processing aids, colorants, sweeteners, flavorings, diluents, and other known additives to provide an aesthetically pleasing presentation of the pharmaceutical product (e.g., the compound of the present invention or a pharmaceutical composition thereof) or to facilitate the preparation of a pharmaceutical product (e.g., a drug).

Examples of suitable oral dosage forms are tablets containing about 1 to 1000 mg of the compound of the present invention combined with about 1 to 1000 mg of anhydrous lactose, about 1 to 1000 mg of croscarmellose sodium, about 1 to 1000 mg of polyvinylpyrrolidone (PVP) K30 and about 1 to 1000 mg of magnesium stearate. The powdered ingredients are first mixed together, and then mixed with a PVP solution. The resulting composition can be dried, granulated, mixed with magnesium stearate, and compressed into tablet form using conventional equipment. Examples of aerosol formulations 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), with the addition of a permeation enhancer (e.g., a salt such as sodium chloride) if desired. The solution can be filtered, for example, using a 0.2-micron filter to remove impurities and contaminants.

Therefore, the embodiments include a pharmaceutical composition comprising a compound of formula (I) or its stereoisomer or pharmaceutical salt. Further embodiments include a pharmaceutical composition comprising a compound of formula (I) or its stereoisomer or pharmaceutical salt and a pharmaceutical carrier or excipient.

Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for treating mutant KRAS-driven cancer.

The following compositions A and B illustrate typical compositions of the present invention, but are only representative examples.

Composition A

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

Composition B

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

Indications and treatment methods

The compounds of this invention induce novel binding pockets in KRAS by driving the formation of a high-affinity triple complex between the KRAS protein and the widely expressed cyclic cyclophilin A (CYPA). These compounds inhibit the interaction of KRAS with downstream effectors such as RAF and PI3K. Therefore, the compounds of this invention can be used to inhibit propagating oncogenic MAPK and PI3K signaling, reducing cell (especially cancer cell) proliferation. The compounds of this invention can be used to terminate RAS signaling in cells expressing RAS mutants (especially KRAS mutants) driving RAS signaling in pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma, ovarian cancer, endometrial cancer, etc. Alternatively, the compounds of this invention can be used to terminate RAS signaling in malignant solid tumors where the oncogenic effects of KRAS mutations are enhanced by dysregulation or mutation of effector pathways such as MAPK and PI3K-AKT-mTOR (a mammalian target of rapamycin). These compounds are used in targeted therapies for pancreatic cancer, colorectal cancer, non-small cell lung cancer, etc.

Another embodiment includes a method for treating or preventing cancer in a mammal in need of such treatment, wherein the method comprises administering to the mammal a therapeutically effective amount of a compound of formula (I), its stereoisomers, tautomers, prodrugs, or pharmaceutical salts.

synthesis

The compounds of this invention can 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 stated, all substituents, in particular ^R1 to ^R6 , ^A1 and ^A2 , are as defined above. Furthermore, unless explicitly stated otherwise, all reactions, reaction conditions, abbreviations and symbols have meanings well known to those skilled in the art of organic chemistry.

The general synthetic route for preparing compound (I) is shown below.

Option 1

Compound (II) is synthesized according to the procedures described in intermediates A through I. Compound (I) can be obtained by coupling acid (III) with compound (II) in the presence of bases such as TEA, DIEPA, and DMAP using coupling agents such as _T3P , HATU, PyBOP, and EDCI/HOBt.

The compounds of the present invention can be obtained in diastereomer or mixture of diastereomers, and they can be separated by methods well known in the art, such as (chiral) HPLC or SFC. In another embodiment, the compounds of formula (I) can be obtained using the corresponding chiral starting materials according to the above scheme.

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

a) Make compound of formula (II),

It undergoes a coupling reaction with acid (III) in the presence of a coupling agent and a base to form compound (I);

in

In step a), the coupling agent may be, for example, _T3P , HATU, PyBOP or EDCI/HOBt; the base may be, for example, TEA, DIEPA or DMAP.

Compounds of formula (I) or (Ia) produced according to the above method are also an object of this invention.

Example

The invention will be more fully understood by referring to the following examples. However, they should not be construed as limiting the scope of the invention.

abbreviation

The invention will be more fully understood by referring to the following examples. However, they should not be construed as limiting the scope of the invention.

The abbreviations used in this article are as follows:

ACN Acetonitrile

aq. water-based

Boc-N-Me-Val-OH N-(tert-butoxycarbonyl)-N-methyl-L-valine

(Boc) _₂O ditert-butyl dicarbonate

(R)-binap (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binathyl

CDCl ₃ : Deuterated chloroform

_CD3 OD: Deuterated methanol

COMU (1-cyano-2-ethoxy-2-oxoethyleneaminooxy)dimethylamino-morpholino-carbomony hexafluorophosphate

DIEPA: N,N-Diethylpropylamine

DIBAL-H diisobutylaluminum hydride

DMAP: 4-Dimethylaminopyridine

DMF: Dimethylformamide

DMP 1,1,1-Tris(acetoxy)-1,1-dihydro-1,2-benzoxanol-3-(1H)-one

DMSO: Dimethyl sulfoxide

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

EtOAc or EA: Ethyl acetate

FRET fluorescence resonance energy transfer

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

hr: hours

HPLC: High Performance Liquid Chromatography

HOBt: N-hydroxybenzotriazole

H-VAL-OTBU HCl (S)-2-amino-3-methylbutyrate tert-butyl hydrochloride

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

LDA (Lithium diisopropylamino)

MS (ESI): Mass spectrometry (electrospray ionization)

min minutes

MTBE (methyl tert-butyl ether)

NMM N-methylmorpholine

NMR: Nuclear Magnetic Resonance

NMO 4-methylmorpholine N-oxide

obsd. Observation value

Pd(dppf) _Cl₂ [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride

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

preparative high performance liquid chromatography (prep-HPLC)

PyBOP: Benzotriazole-1-yloxytripyrrolidinephosphine hexafluorophosphate

RT or rt: room temperature

sat. saturated

SFC Supercritical Fluid Chromatography

TEA: Triethylamine

TFA: Trifluoroacetic acid

THF: Tetrahydrofuran

TEA: Trimethylamine

TMEDA Tetramethylethylenediamine

TMSCF ₃ trifluoromethyltrimethylsilane

T ₃ P: Propylphosphonic anhydride

General experimental conditions

Purify intermediates and final compounds using rapid chromatography with 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 Registry Number: 63231-67-4, particle size: 47-60 μm silica gel; iii) ZCX from Qingdao Ocean Chemical Co., Ltd., pore size: 200-300 or 300-400.

Intermediates and final compounds were purified by preparative HPLC on reversed-phase columns using XBridge ^™ Prep-C18 (5 μm, OBD™ 30 × 100 mm), SunFire ^™ Prep-C18 (5 μm, OBD ^™ 30 × 100 mm), Phenomenex Synergi-C18 (10 μm, 25 × 150 mm), or Phenomenex Gemini-C18 (10 μm, 25 × 150 mm). A Waters AutoP purification system (sample manager 2767, pump 2525, detectors: 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) was used. Alternatively, a 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).

For chiral separation of SFC, intermediates were separated using chiral columns (Daicel chiralpak IC, 5 μm, 30 × 250 mm), AS (10 μm, 30 × 250 mm), or AD (10 μm, 30 × 250 mm), employing the Mettler Toledo Multigram III SFC system, Waters 80Q preparative SFC, or Thar 80 preparative SFC. Solvent systems were _CO2 and IPA (0.5% TEA in IPA solution) or _CO2 and MeOH (0.1% _NH3 · _H2O in MeOH solution), with a back pressure of 100 bar. UV detection was performed at 254 or 220 nm.

The LC/MS spectra of the compounds were obtained using LC/MS (Waters ^™ Alliance 2795-Micromass ZQ, Shimadzu Alliance 2020-Micromass ZQ, or Agilent Alliance 6110-Micromass ZQ) under the following conditions (run time 3 or 1.5 minutes):

Acidic conditions I: A: 0.1% TFA solution in _H₂O ; B: 0.1% TFA solution in acetonitrile;

Acidic conditions II: A: 0.0375% TFA solution in _H₂O ; B: 0.01875% TFA solution in acetonitrile;

Alkaline conditions I: A: 0.1% _NH3 · _H2O solution in _H2O ; B: Acetonitrile;

Alkaline conditions II: A: 0.025% _NH3 · _H2O solution in _H2O ; B: Acetonitrile;

Neutral conditions: A: _H₂O ; B: Acetonitrile.

Mass spectrometry (MS): Usually only ions representing the parent mass are reported, unless otherwise specified, the mass ions cited are positive mass ions (MH) ⁺ .

NMR spectra were obtained using a Bruker Avance at 400 MHz or 500 MHz.

Microwave-assisted reactions were performed in a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were carried out under an argon or nitrogen atmosphere. Unless otherwise specified, reagents were purchased as is from commercial suppliers without further purification.

Preparation Examples

Preparation of intermediates

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

Title intermediate A was prepared according to the following scheme:

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

Add 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) to a solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (compound A1, 2.0 g, 9.26 mmol) and bis(pinacol)diboron (3.5 g, 13.9 mmol) in THF (30 mL). Stir the mixture at 75 °C for 16 hours under _N₂ . Filter the mixture and concentrate the filtrate under vacuum. 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-dioxaneborane-2-yl)pyridine (compound A2, 2.4 g), which was a yellow oil. ^¹H NMR (400 MHz, CDCl₃) δppm 8.91 (d, J = 1.4 Hz, 1H), 8.21 (d, J = 1.4 Hz, 1H), 4.95 (q, J = 6.5 Hz, 1H), 3.30 (s, 3H), 1.49 (d, J = 6.5 Hz, 3H), 1.35 (s, 12H).

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

N-iodosuccinimide (4.1 g, 18.27 mmol) was added to a solution of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron-2-yl)pyridine (compound A2, 2.5 g, 7.3 mmol) in ACN (40 mL). The mixture was stirred at 90 °C for 40 h under _N2 protection. The reaction was quenched with saturated _Na2SO3 solution ( ₄₀ mL), and the reaction mixture was extracted with EtOAc (30 mL, twice). The combined organic layers were washed with brine (50 mL), filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (compound A3, 660 mg) as a yellow oil. MS calculated value 342 (MH ⁺ ); measured value 341.8 (MH ⁺ ).

Step 3: Preparation of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound A5)

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) were added 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). The mixture was stirred at 100 °C for 12 hours under _N2 . 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 benzyl 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound A5, 740 mg) as a yellow solid. MS calculated value 434.1 (MH ⁺ ); measured value 434.1 (MH ⁺ ).

Step 4: Preparation of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-3-pyridyl]-4-methyl-piperazine (intermediate A)

KOAc (418.0 mg, 4.26 mmol) and Pd(dppf)Cl₂ (124.7 mg, 0.170 mmol) were added to a solution of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound A5, 740 mg, 1.7 mmol) and bis(pinacol) _diboron (519.2 mg, 2.04 mmol) in toluene (12 mL). The reaction mixture was stirred at 90 °C for 12 hours under _N₂ protection. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography to give 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-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 ⁺ ).

Intermediate B

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

Intermediate B was prepared according to the following scheme:

Step 1: Preparation of (4-bromothiazol-2-yl)methanol (compound B2)

Sodium borohydride (1.7 g, 46.87 mmol) was added to a solution of 4-bromothiazol-2-carboxaldehyde (compound B1, 6.0 g, 31.25 mmol) in methanol (70 mL) at 0 °C. The mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (300 mL) at 0 °C, and the reaction mixture was extracted 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.

Step 2: Preparation of 4-bromo-2-(bromomethyl)thiazole (compound B3)

At 0 °C, _CBr₄ (15.4 g, 46.38 mmol) and triphenylphosphine (12.1 g, 46.38 mmol) were added to a solution of (4-bromothiazol-2-yl)methanol (compound B2, 6.0 g, 30.92 mmol) in DCM (80 mL). 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 chromatography, 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⁺ ).

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

At -78 °C, n-butyllithium (10 mL, 25.22 mmol, 2.5 M) was slowly added 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). After addition, the mixture was stirred at -78 °C for 0.5 h. At -78 °C, 4-bromo-2-(bromomethyl)thiazole (compound B3, 5.4 g, 21.02 mmol) was added to the above mixture, and the mixture was stirred for another 1 h. The reaction was quenched with saturated _NH4Cl solution (100 mL), and the reaction mixture was extracted with EtOAc (100 mL, twice). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by reversed-phase chromatography to give 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (compound B5, 3.6 g), a yellow oil. MS calculated value 360 (MH ⁺ ); measured value 359.9 (MH ⁺ ).

Step 4: Preparation of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound B6)

Hydrochloric acid (66.6 mL, 0.3 M) was added 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). The mixture was stirred at 25 °C for 2 hours. The mixture was alkalized with saturated _NaHCO3 solution 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 methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound B6, 3.1 g) as a yellow oil. MS calculated value 264.9 (MH ⁺ ); measured value 264.9 (MH ⁺ ).

Step 5: Preparation of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound B7)

Triethylamine (2.9 g, 29.23 mmol) and (Boc) _₂O (3.8 g, 17.54 mmol) were added to a solution of (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound B6, 3.1 g, 11.69 mmol) in DCM (40 mL). After stirring at 30 °C for 12 hours, the mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate (0–30%) in petroleum ether to give methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound B7, 3.2 g) as a yellow oil. MS calculated value 387 ( ^MNa⁺ ); measured value 386.9 ( ^MNa⁺ ).

Step 6: Preparation of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionic acid (compound B8)

Lithium hydroxide (0.4 mL, 43.81 mmol) was added to a solution of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound B7, 3.2 g, 8.76 mmol) in THF (30 mL), methanol (2 mL), and water (10 mL). After stirring at 25 °C for 1 hour, the reaction mixture was acidified with 1 M 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)propionate (compound B8, 3.1 g) as a yellow oil. MS calculated value 373 (MNa ⁺ ); measured value 372.9 (MNa ⁺ ).

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

At 0 °C, methyl (3S)-hexahydropyridazine-3-carboxylate, (compound B8, 3.1 g, 8.83 mmol), EDCI (3.4 g, 17.65 mmol), 1-hydroxybenzotriazole (238.5 mg, 1.77 mmol), and NMM (9.92 mL, 88.26 mmol) were added to a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionic acid (compound B8, 3.1 g, 8.83 mmol) in DCM (50 mL). 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 chromatography and eluted with ethyl acetate (10–30%) in petroleum ether to give (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate B, 2.4 g). MS calculated value 477 (MH ⁺ ), measured value 476.9 (MH ⁺ ).

Intermediate C

(7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione

Title intermediate C was prepared according to the following scheme:

Step 1: Preparation of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropane-1-one (compound C3)

_SnCl₄ solution (97.2 mL, 121.5 mmol) was slowly added to a mixture of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropionyl chloride (compound C1, 35.0 g, 116.8 mmol) in DCM (400 mL) at 0 °C. After stirring at -40 °C for 0.5 hours, a DCM solution of 5-bromo-6-fluoro-1H-indole (compound C2, 25.0 g, 116.8 mmol) in 200 mL was added dropwise to the mixture stirred at -40 °C for 15 min. After the reaction was complete, _the mixture was quenched with saturated _NaHCO₃ aqueous solution (800 mL), and the reaction mixture was extracted with EtOAc (900 mL, twice). The combined organic layers were washed with brine (700 mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was ground with solution (100 mL, petroleum ether: ethyl acetate = 8:1) and filtered. The filter cake was dried under vacuum to give a yellow solid, 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropane-1-one (compound C3, 50.0 g). MS calculated value 552.1 (MH ⁺ ); measured value 552.1 (MH ⁺ ).

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

LiBH₄ (48.4 mL, 193.49 mmol, 4 M THF solution) was added dropwise to a mixture of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropane- _1- one (compound C3, 50.0 g, 90.49 mmol) in THF (600 mL) at 0 °C. The mixture was stirred at 70 °C for 24 h under a nitrogen atmosphere. After the reaction was complete, the reaction was quenched at 0 °C by the slow addition of water (600 mL), and the reaction mixture was extracted twice with EtOAc (600 mL). The combined organic layers were washed with brine (600 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography to give [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 ⁺ ).

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)

Silver trifluoromethanesulfonate (20.3 g, 78.88 mmol) was added 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. The mixture was stirred at 0 °C for 10 min. After the reaction was complete, the reaction was quenched with saturated aqueous solution of _Na₂SO₃ (400 mL) and EtOAc (400 mL), and the reaction mixture was filtered. The organic layer was washed with brine (100 _mL ), dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography to give a yellow solid [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound C5, 43.0 g). MS calculated value 664.0 (MH ⁺ ); measured value 664.1 (MH ⁺ ).

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-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C6)

Potassium phosphate (15.7 g, 73.92 mmol) and Pd(dppf)Cl₂ (920 mg, 1.26 mmol) were added 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-dioxane-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). The mixture was stirred at 70 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the mixture was filtered and concentrated under vacuum. The residue was purified by silica gel column chromatography (EtOAc in PE = 20%–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, 19.5 g) as a white solid. MS calculated value 891.3 (MH ⁺ ); measured value 891.3 (MH ⁺ ).

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-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C7)

2,2,2-trifluoroethyl trifluoromethanesulfonate (37.7 g, 162.56 mmol) was added dropwise to a solution 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-pyridyl]piperazine- ₁ -carboxylate (compound C6, 14.5 g, 16.26 mmol) and _Cs₂CO₃ (15.9 g, 48.77 mmol) in DMF (200 mL), and the mixture was stirred at 20 °C for 12 hours. After the reaction was complete, EtOAc (70 mL) and water (100 mL) were added and the layers were separated. The aqueous phase was extracted twice with EtOAc (70 mL). The combined organic layers were washed with brine (100 mL, four times), dried over _{Na₂SO₄ ,} filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography to give a yellow oil, 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-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C7, 8.0 g, PEAK 1, fast elution). MS calculated value 973.3 (MH ⁺ ); measured value 973.2 (MH ⁺ ).

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-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C8)

Cesium fluoride (8.2 g, 53.9 mmol) was added 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-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C7, 10.5 g, 10.78 mmol) in DMF (130 mL), and the mixture was stirred at 60 °C for 24 h. 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), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give the residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 25%–66%) to give a yellow solid, 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-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C8, 6.5 g). MS calculated value 735.2 (MH ⁺ ); measured value 735.1 (MH ⁺ ).

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-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C9)

Pd(dppf)Cl₂ (537.1 mg, 0.73 mmol) was added to a solution 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-pyridyl]piperazine-1-carboxylate (compound _C8 , 5.4 g), bis(pinacol)diboron (2.8 g, 11.01 mmol), and potassium acetate (1.2 mL, 18.35 mmol) in toluene (70 mL). The mixture was degassed and purged three times under a nitrogen atmosphere, and stirred at 90 °C for 12 hours. After the reaction was complete, the mixture was cooled to room temperature. The reaction mixture was filtered, and the filtrate was concentrated under vacuum to give the residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 25%–66%) to give a yellow oily substance, 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C9, 5.2 g). MS calculated value 783.3 (MH ⁺ ); measured value 783.3 (MH ⁺ ).

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]thiazolyl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound C10)

Under a nitrogen atmosphere, K₃PO₄ was added to a mixture of (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate B, 2.7 g, 5.69 mmol) and 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound _C9 , 4.9 g, 6.32 mmol) in toluene (60 mL)/1,4-dioxane (20 mL)/water (20 mL ₎ . (3.4 g, 15.81 mmol) and Pd(dtbpf) _Cl₂ (412.2 mg, 0.63 mmol). The mixture was stirred at 70 °C for 12 hours. After the reaction was complete, the mixture was concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 10%–75%) to give methyl (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]thiazolyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (compound C10, 3.6 g). MS calculated value 1053.4 (MH ⁺ ); measured value 1053.3 (MH ⁺ ).

Step 9: 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]thiazolyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid (compound C11)

To a solution 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]thiazolyl]-2-(tert-butoxycarbonylamino)-propionyl]-hexahydropyridazine-3-carboxylate (compound C10, 3.6 g, 3.42 mmol) in a DCE (50 mL), trimethyltin alcohol (2.4 g, 13.67 mmol) was added and the mixture was stirred at 60 °C for 12 h. After the reaction was complete, EtOAc (80 mL) and water (60 mL) were added and the layers were separated. The aqueous phase was extracted twice with EtOAc (80 mL). The combined organic layers were washed with brine (100 mL), dried over _Na₂SO₄ , 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]thiazolyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid (compound C11, 4.3 g), a brown solid. MS calculated value 1039.4 (MH ⁺ ); measured value 1039.2 (MH ⁺ ).

Step 10: Preparation of 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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C12)

DIEA (14.4 mL, 82.76 mmol), EDCI (11.9 g, 62.07 mmol), and 1-hydroxybenzotriazole (1.4 g, 10.35 mmol) were added 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]thiazolyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid (compound C11, 4.3 g, 4.14 mmol) in DCM (430 mL) at 0 °C. The mixture was stirred at 15 °C for 12 hours. After the reaction was complete, the mixture was concentrated under vacuum, then diluted with water (80 mL), and extracted with EtOAc (80 mL, twice). The combined organic layers were washed with brine (80 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (EtOAc in PE = 25%–66%) to obtain a yellow gel-like substance, 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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C12, 3.1 g). MS calculated value 1021.4 (MH ⁺ ); measured value 1021.2 (MH ⁺ ).

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

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]} A mixture of 1,8-octadecano-1(25),2,5(28),19,22(26),23-hexaden-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C12, 3.1 g, 3.04 mmol) and an aqueous solution of formaldehyde (775.0 mg, 9.55 mmol) in methanol (150 mL) was mixed with Pd(OH) _₂ on activated carbon (2.79 g, 3.97 mmol). The mixture was degassed and purged three times with _H₂ . The mixture was hydrogenated at 30 °C for 18 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated under vacuum to obtain N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound C13, 2.6 g), which was a brown solid. MS calculated value 901.3 (MH ⁺ ); measured value 901.3 (MH ⁺ ).

Step 12: Preparation of (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate C)

TFA (14.0 mL, 181.72 mmol) was added to a mixture of N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound C13, 2.6 g, 2.89 mmol) in DCM (18 mL). The mixture was stirred at 15°C for 0.5 h. After the reaction was complete, the mixture was concentrated under vacuum and diluted with saturated _NaHCO3 (30 mL), and extracted with EtOAc (30 mL, three times). The combined organic layers were washed with brine (50 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a yellow solid (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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} ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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 ⁺ )

Intermediate D

(7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione

Similar to the preparation of intermediate C, the title compound was prepared by using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate.

Intermediate E

(7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione

The compound was prepared according to the following scheme:

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

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), _Cs₂CO₃ ( _4.85 g, 14.88 mmol), (R)-binap (92.6 mg, 0.15 mmol), and Pd(OAc) _₂ (66.8 mg, 0.3 mmol) were added. The reaction mixture was degassed and purged three times with nitrogen, 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 under vacuum to obtain the residue. The residue was purified by column chromatography to a yellow oil, 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (compound E2, 2.0 g). MS: calculated value 382.2 (MH ⁺ ); measured value 382.1 (MH ⁺ ).

Step 2: 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (compound E3).

Pd(dppf)Cl₂ (306.3 mg, 0.42 mmol) was added to a solution of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (compound E₂, 3.2 g, 8.37 mmol), bis(pinacol)diboron (3.19 g, 12.56 mmol), and KOAc (2.1 g, 20.93 mmol) in _toluene (50 mL). The mixture was degassed and purged three times with nitrogen, and then 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 under vacuum to obtain a residue, which was purified by reverse-phase column chromatography to give a yellow gel-like substance, 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (compound E3, _1.9 g). MS calculated value 430.2 (MH ⁺ ); measured value 348.4 (MC _6H10 +H ⁺ ).

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

To a solution of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (compound E3, 1.9 g, 4.41 mmol) and [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), _K₃PO₄ ( _2.1 g, 9.5 mmol) and Pd(dppf) _Cl₂ (231 mg, 0.37 mmol) were added. The mixture was degassed by purging nitrogen for 2 min, and the reaction mixture was stirred at 70 °C for 12 h. After cooling to room temperature, the reaction mixture was filtered. The filtrate was concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (EtOAc in PE: 30%–60%) to give a yellow gel-like substance, [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound E4, 960.0 mg). MS: calculated value 839.3 (MH ⁺ ); measured value 839.3 (MH ⁺ ).

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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound E5).

Cs₂CO₃ (1.1 g, 3.44 mmol) and 2,2,2-trifluoroethyl trifluoromethanesulfonic acid (2.7 g, 11.63 mmol) were added to a solution of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1H-indol- ₃ -yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound _E4 , 1 g, 1.14 mmol) in DMF (35 mL) at 0 °C. 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 organics were washed with brine (50 mL, three times), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc in PE: 30%–40%) to give a white solid [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound E5, 640.0 mg, rapid elution). MS calculated value 921.3 (MH ⁺ ); measured value 921.4 (MH ⁺ ).

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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propane-1-ol (compound E6).

Cesium fluoride (421.8 mg, 2.78 mmol) was added 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-pyridyl]-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). 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 under vacuum to obtain the residue. The residue was purified by column chromatography (EtOAc in PE: 30%–60%) to give a yellow oily substance, 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propane-1-ol (compound E6, 360.0 mg). MS calculated value 683.2 (MH ⁺ ); measured value 683.1 (MH ⁺ ).

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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propane-1-ol (compound E7).

Potassium acetate (0.08 mL, 1.32 mmol) and Pd(dppf)Cl₂ (40 mg, 0.1 mmol) were added 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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propane-1-ol (compound E6, 360.0 mg, 0.53 mmol), bis(pinacol)diboron (200.6 mg, 0.79 mmol), and _toluene (6 mL). The reaction mixture was degassed by purging nitrogen for 5 min and then stirred at 80 °C for 15 h. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (EtOAc in PE: 30%–50%) to give a yellow gelatinous substance, 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propane-1-ol (compound E7, 300.0 mg). MS calculated value 731.4 (MH ⁺ ); measured value 731.4 (MH ⁺ ).

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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound E8).

Add K₃PO₄ (221.3 mg, 1.04 mmol) and Pd(dtbpf)Cl 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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propane-1-ol (compound E7, 0.3 g, 0.41 mmol) and (3S)-1-[(2S) _-3- (4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate B, 196.7 _mg , 0.41 mmol) in toluene (3 mL), 1,4-dioxane (1 mL), and water (1 mL). ₂ (27.05 mg, 0.04 mmol). 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 under vacuum to give the residue. The residue was purified by column chromatography (EtOAc in PE: 60%–80%) to give a yellow gel-like (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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound E8, 200.0 mg). MS calculated value 1001.4 (MH ⁺ ); measured value 1001.4 (MH ⁺ ).

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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazin-3-carboxylic acid (compound E9).

Me 3 SnOH (200.0 mg, 1.11 mmol) was added 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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine- ₃ -carboxylate (compound E8, 200.0 mg, 0.2 mmol) in DCE (5 mL). The mixture was stirred at 60 °C for 12 hours. The reaction mixture was concentrated under vacuum to obtain a residue. EtOAc (10 mL) and water (10 mL) were added to the residue, and the layers were separated. The aqueous phase was extracted twice with EtOAc (15 mL). The combined organic layers were washed with brine (20 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum 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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazin-3-carboxylic acid (compound E9, 188.0 mg) as a brown solid. MS calculated value 987.4 (MH ⁺ ); measured value 987.4 (MH ⁺ ).

Step 9: Preparation of N-[(7S,13S)-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]tert-butyl carbamate (compound E10).

At 0 °C, DIEA (0.7 mL, 3.81 mmol), EDCI (550.0 mg, 2.87 mmol), and HOBt (65.0 mg, 0.48 mmol) were added 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-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazin-3-carboxylic acid (compound E9, 188.0 mg, 0.19 mmol) in DCM (20 mL). 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), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc in PE: 50%-70%) to give a yellow solid N-[(7S,13S)-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-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 [C18-1(25),2,5(28),19,22(26),23-hexene-7-yl]tert-butyl carbamate (compound E10, 110.0 mg). MS calculated value 969.4 (MH ⁺ ); measured value 969.5 (MH ⁺ ).

Step 10: Preparation of (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-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 ]-octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E).

To N-[(7S,13S)-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-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 [C28C-1(25),2,5(28),19,22(26),23-hexene-7-yl]tert-butyl carbamate (compound E10, 110.0 mg, 0.11 mmol) was added to a solution of TFA (1.0 mL, 12.98 mmol) in DCM (1 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was concentrated under vacuum to obtain a residue. A saturated aqueous solution of _NaHCO3 (20 mL) was added, 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 under vacuum to obtain a yellow solid (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E, 98.0 mg). MS calculated value 869.4 (MH ⁺ ); measured value 869.2 (MH ⁺ ).

intermediate F

(7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione

Similar to the preparation of intermediate E, the title compound was prepared by using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate.

intermediate G

(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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione

Similar to the preparation of intermediate E, the title compound was prepared by using iodoethane and morpholine instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate and 1-(2,2,2-trifluoroethyl)piperazine (compound E1).

intermediate H

(7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione

Similar to the preparation of intermediate C, the title compound was prepared by replacing 5-bromo-6-fluoro-1H-indole (compound C2) with 5-bromo-4-fluoro-1H-indole.

Intermediate I

(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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione

Similar to the preparation of intermediate E, the title compound was prepared by using morpholine instead of 1-(2,2,2-trifluoroethyl)piperazine (compound E1).

Example 1

(1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropaneformamide

At 0 °C, 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-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 ] were added to a solution of (1S,2S)-2-methylcyclopropanecarboxylic acid (compound 1a, 64.3 mg, ^{0.64 mmol} ) in DMF (2 mL). [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-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), _dried over _Na₂SO₄ , and concentrated under vacuum to give the residue.] The resulting residue was purified by silica gel chromatography to give (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropanecarboxamide as a white solid (Example 1, 156.41 mg). MS calculated value 829.5 (MH ⁺ ); measured value 829.5 (MH ⁺ ). ¹ H NMR (400MHz, 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.

Example 2

(1R,5S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-oxabicyclo[3.1.0]hexane-6-carboxamide

Similar to the preparation of Example 1, the title compound was prepared by replacing (1S,2S)-2-methylcyclopropanecarboxylic acid (compound 1a) with (1S,5R)-3-oxabicyclo[3.1.0]hexane-6-carboxylic acid. 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 (400MHz, 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.

Example 3

(1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]cyclopropaneformamide

Similar to the preparation in Example 1, the title compound was prepared 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 ⁺ ). ^¹H NMR (400 MHz, chloroform-d) δ＝8.76 (br d, J＝2.6 Hz, 1H), 8.66 (d, J＝7.6 Hz, 1H), 7.63 (d, J＝2.1 Hz, 1H), 7.30 (br d, J＝2.4 Hz, 1H), 7.12 (d, J＝12.3 Hz, 1H), 6.78 (br d, J＝2.4 Hz, 1H), 6.12 (br ...12 (br d, J＝12.3 Hz, 1H), 6.12 (br d, J＝12.3 Hz, 1H), 6.12 ( 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.

Example 4

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

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 The title compound was prepared by replacing (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D) with octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione. Example 4 (15.7 mg) was obtained as a white solid. MS calculated value 883.5 (MH ⁺ ); measured value 883.1 (MH ⁺ ). ^¹H NMR (400MHz, methanol- _d⁴ ) 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), 5.71(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(m, 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

-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).

Examples 5 and 6

(1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexen-7-yl]cyclopropaneformamide and (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]octadecane-1(25),2,5(28),19,22(26),23-hexen-7-yl]cyclopropaneformamide

The compound was prepared according to the following scheme:

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-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 Preparation of cyclopropane formamide and (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-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 ]cyclopropane formamide

At 0 °C, a solution of trans-2-cyanocyclopropanecarboxylic acid (compound 5a, 11.2 mg, 0.1 mmol) in DMF (0.5 mL) was treated with 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-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 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-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, fast elution) and Example 6 (9.9 mg, slow elution) as white solids.]

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.

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.

Example 7

(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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropaneformamide

Similar to the preparation in Example 1, this was achieved 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-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 The title compound was prepared by replacing (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 7 (12.5 mg) was obtained as a yellow solid. MS calculated value 897.4 (MH ⁺ ); measured value 897.6 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )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.

Example 8

(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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropaneformamide

Similar to the preparation in Example 1, this was achieved 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 The title compound was prepared by replacing (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 8 (135.4 mg) was obtained as a pale yellow solid. MS calculated value 816.4 (MH ⁺ ); measured value 816.1 (MH ⁺ ). ¹ H NMR (400MHz, 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.

Example 9

(1S,2S)-N-[(7S,13S)-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropaneformamide

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-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 The title compound was prepared by replacing (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 9 (27.7 mg) was obtained as a grayish-white solid. MS calculated value 951.4 (MH ⁺ ); measured value 951.2 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )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.

Example 10

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

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 The title compound was prepared by replacing (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 10 (11.2 mg) was obtained as a white solid. MS calculated value 883.4 (MH ⁺ ); measured value 883.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ＝8.50 (d, J＝2.8 Hz, 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.

Example 11

(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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]cyclopropaneformamide

Similar to the preparation in Example 1, this was achieved 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 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate G) and (1S,2S)-2-(difluoromethyl)cyclopropanecarboxylic acid replacing (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D) and (1S,2S)-2-methylcyclopropanecarboxylic acid (compound 1a). Example 11 (33.2 mg) was obtained as a yellow solid. MS calculated value 852.4 (MH ⁺ ); measured value 852.2 (MH ⁺ ). ¹ H 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.

Example 12

(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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropaneformamide

Similar to the preparation in Example 1, this was achieved 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 The title compound was prepared by replacing (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-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 ]octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D) with octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione. Example 12 (150 mg) was obtained as a white solid. MS calculated value 870.4 (MH ⁺ ); measured value 870.5 (MH ⁺ ). ^¹H NMR (400MHz, methanol- _d⁴ ) 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,1H), 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,1H). 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.

Biological examples

Compound A122 from WO2022060836 (page 70 of Table 1a) is cited as a reference compound in this invention.

(A122)

Example 13

Cell viability assay

The purpose of this cell assay was to determine the effect of the tested compound on the proliferation of human cancer cell lines NCI-H358 (ATCC-CRL5807), AGS (ATCC-CRL-1739), and 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.

Cells were seeded at 5,000 cells/well (NCI-H358), 2,000 cells/well (AGS), and 2,000 cells/well (SW620) in 96-well assay plates (Corning-3699) and incubated overnight. On the day of assay, diluted compound was added at a final concentration of 0.5% DMSO. After 72 hours of incubation, one-tenth volume of Cell Counting Kit 8 (Dnjindo-CK04) was added to each well. After 2 hours of incubation, the signal (OD450 minus OD650) was read using EnVision. _IC50 was determined by fitting a 4-parameter sigmoid concentration response model.

Table 1. Examples of the present invention and the activity of compounds in KRAS cell viability assay

Example 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

Example 14

Interaction determination of KRAS-BRAF with CYPA (500 nM)

In this example, TR-FRET was also used to measure the compound- or compound-CYPA-dependent disruption of the KRAS G12C-BRAF complex. This protocol was also used to measure the disruption of KRAS G12D or KRAS G12V binding to BRAF by the compounds of this invention, respectively. The solutions were mixed in the wells of a 384-well assay plate containing 25 mM HEPES (pH 7.4, 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid, Thermo, 15630080), 0.002% Tween 20, 0.1% BSA, 100 mM NaCl, ₅ mM MgCl₂, 10 μM GMPPNP (guanosine 5'-[β,γ-imino]trisodium triphosphate hydrate, Sigma, G0635), untagged CYPA, GMPPNP loaded with 6His-KRAS protein, and GST-BRAF ^RBD at final concentrations of 50 nM, 6.25 nM, and 1 nM, respectively. Compounds were introduced into wells at a final concentration of 10 μM in a 16-point, 3-fold dilution series and incubated for 3 hours. Then, mixtures of MAb anti-6His-XL665 (Cisbio, 61HISXLB) and Mab anti-GST-TB well compound (Cisbio, 61GSTTLB) were added at final concentrations of 6.67 nM and 0.21 nM, respectively, and the plates were incubated for another 1.5 hours. TR-FRET signals were read on a PHERstar FSX microplate reader (Ex 320 nm, Em 665/615 nm). Compounds that promoted the disruption of the KRAS-BRAF complex were identified as those causing a decrease in TR-FRET ratios compared to the DMSO control wells.

Table 2. Examples of the present invention and the activity of compounds in the KRAS-BRAF-CYPA (500 nM) interaction assay.

Example <![CDATA[G12C IC₅₀(μM)]]> <![CDATA[G12D IC₅₀(μM)]]> <![CDATA[G12V IC₅₀(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

Example 15

pERK inhibition assay

This assay aimed to measure the ability of the tested compounds to inhibit ERK phosphorylation, KRAS G12C in NCI-H358 cells, KRAS G12D in AGS cells, and downstream signaling of KRAS G12V in SW620 cells. 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 adding the compounds, cells were seeded at densities of 30,000 cells/well, 20,000 cells/well, and 30,000 cells/well (for NCI-H358, AGS, and SW620) in tissue culture-treated 96-well plates (Corning-3699) and allowed to adhere overnight. The diluted compound was then added to a final concentration of 0.5% DMSO. After incubation for 4 hours, the medium was removed, 100 μL of 4% formaldehyde was added, and the plate was incubated at room temperature for 20 minutes. The plate was then washed once with phosphate-buffered saline (PBS) and permeated with 100 μL of frozen methanol for 10 minutes. Non-specific antibodies binding to the plate were blocked for at least 1 hour at room temperature using 50 μL of 1X BSA blocking buffer (Thermo-37520, diluted 10-fold with phosphate-buffered saline (PBST)).

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

IC ₅₀ was determined by fitting a 4-parameter S-type concentration response model.

Table 3. Examples of the present invention and the activity of compounds in KRAS pERK inhibition assays.

Example <![CDATA[G12C IC₅₀(μM)]]> <![CDATA[G12D IC₅₀(μM)]]> <![CDATA[G12V IC₅₀(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 (29)

1. A compound of formula (I), in R ¹ is 3-oxabicyclo[3.1.0]hexyl, _C1-6 alkyl _C3-7 cycloalkyl, cyano _C3-7 cycloalkyl, or halo- _C1-6 alkyl _C3-7 cycloalkyl; ^R2 is H or a halogen; ^R3 is H or a halogen; ^R4 is a _C1-6 alkyl or a halo- _C1-6 alkyl; R ⁵ is a _C1-6 alkoxy- _C1-6 alkyl group; ^R6 is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino; ^A1 is an imidazolyl group; ^A2 is a _C1-6 alkylene group; The condition is that ^R2 and ^R3 are not both H at the same time; Or its medicinal salt. 2. A compound of formula (Ia), in R ¹ is 3-oxabicyclo[3.1.0]hexyl, _C1-6 alkyl _C3-7 cycloalkyl, cyano _C3-7 cycloalkyl, or halo- _C1-6 alkyl _C3-7 cycloalkyl; ^R2 is H or a halogen; ^R3 is H or a halogen; ^R4 is a _C1-6 alkyl or a halo- _C1-6 alkyl; R ⁵ is a _C1-6 alkoxy- _C1-6 alkyl group; ^R6 is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino; ^A1 is an imidazolyl group; ^A2 is a _C1-6 alkylene group; The condition is that ^R2 and ^R3 are not both H at the same time; Or its medicinal salt. 3. The compound according to claim 1 or 2, wherein ^R1 is a _C1-6 alkyl- _C3-7 cycloalkyl or a halogenated _C1-6 alkyl _-C3-7 cycloalkyl. 4. The compound according to any one of claims 1 to 3, wherein ^R1 is methylcyclopropyl or (difluoromethyl)cyclopropyl. 5. The compound according to any one of claims 1 to 4, wherein ^R2 is H or fluorine. 6. The compound according to any one of claims 1 to 5, wherein ^R3 is H or fluorine. 7. The compound according to any one of claims 1 to 6, wherein ^R4 is ethyl or 2,2,2-trifluoroethyl. 8. The compound according to any one of claims 1 to 7, wherein R ⁵ is 1-methoxyethyl. 9. The compound according to any one of claims 1 to 8, wherein ^R6 is morpholino or _C1-6 alkylpiperazinyl. 10. The compound according to any one of claims 1 to 9, wherein R ⁶ is morpholino or 4-methylpiperazin-1-yl. 11. The compound according to any one of claims 1 to 10, wherein ^A1 is a compound in which bond "a" is attached to an indole ring. 12. The compound according to any one of claims 1 to 11, wherein ^A2 is 2,2-propylidene. 13. The compound according to claim 1 or 2, wherein... ^R1 is a _C1-6 alkyl- _C3-7 cycloalkyl or a halogenated _C1-6 alkyl- _C3-7 cycloalkyl; ^R2 is H or a halogen; ^R3 is H or a halogen; ^R4 is a _C1-6 alkyl or a halo- _C1-6 alkyl; R ⁵ is a _C1-6 alkoxy- _C1-6 alkyl group; ^R6 is morpholino or _C1-6 alkylpiperazino; ^A1 is where the "a" bond is attached to the indole ring; ^A2 is a _C1-6 alkylene group; The condition is that ^R2 and ^R3 are not both H at the same time; Or its medicinal salt. 14. The compound according to claim 13, wherein... ^R1 is 2-methylcyclopropyl or 2-(difluoromethyl)cyclopropyl; ^R2 is H or fluorine; ^R3 is H or fluorine; ^R4 is ethyl or 2,2,2-trifluoroethyl; R ⁵ is (1S)-1-methoxyethyl; ^R6 is morpholino or 4-methylpiperazin-1-yl; ^A1 is where the "a" bond is attached to the indole ring; ^A2 is 2,2-propylidene; The condition is that ^R2 and ^R3 are not both H at the same time; Or its medicinal salt. 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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexene-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]cyclopropaneformamide; (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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-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 [Octadecano-1(25),2,5(28),19,22(26),23-hexene-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-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] [Octadecano-1(25),2,5(28),19,22(26),23-hexene-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] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-2-methyl-cyclopropaneformamide; (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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]cyclopropaneformamide; 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 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-2-methyl-cyclopropanecarboxamide; Or its medicinal salt. 16. A method for preparing the compound according to any one of claims 1 to 15, the method comprising any one of the following steps: a) Make compound of formula (II), It undergoes a coupling reaction with acid (III) in the presence of a coupling agent and a base to form the compound of formula (I); ^R1 , ^R2 , ^R3 , ^R4 , ^R5 , ^R6 , ^R7 , ^A1 , and ^A2 are as defined in any one of claims 1 to 14; the coupling agent is _T3P , HATU, PyBOP, or EDCI/HOBt; and the base is TEA, DIEPA, or DMAP. 17. The compound or pharmaceutical salt according to any one of claims 1 to 15, which is used as a therapeutically active substance. 18. A pharmaceutical composition comprising the compound according to any one of claims 1 to 15 and a pharmaceutical excipient. 19. Use of the compound according to any one of claims 1 to 15 for the treatment of KRAS G12C protein-related diseases. 20. Use of the compound according to any one of claims 1 to 15 for the treatment of diseases related to KRASG12C, G12D and G12V proteins. 21. Use of the compound according to any one of claims 1 to 15 for suppressing the interaction between RAS and downstream effectors, wherein the downstream effectors are RAF and PI3K. 22. Use of the compound according to any one of claims 1 to 15 for inhibiting the transmission of carcinogenic MAPK and PI3K signaling. 23. Use of the compound according to any one of claims 1 to 15 for the treatment or prevention of KRAS mutation-driven cancers, wherein said cancers are selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma, ovarian cancer, and endometrial cancer. 24. Use of the compound according to any one of claims 1 to 15 for the treatment or prevention of KRAS mutation-driven cancers, wherein said cancers are selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer. 25. The compound or pharmaceutical salt according to any one of claims 1 to 15, for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer. 26. Use of the compound according to any one of claims 1 to 15 for the preparation of a medicament for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer. 27. A method for treating or preventing 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. The compound or pharmaceutical salt according to any one of claims 1 to 15, prepared according to the method of claim 16. 29. The invention as described in the specification.