Macrocyclic inhibitors of KRAS for the treatment of cancer

Abstract

The present invention relates to a compound of formula (I) TIFF2025522629000100.tif77170(wherein R 1 to R 7 , A 1 and A 2 are as described herein) and their pharmaceutically acceptable salts, as well as compositions containing the compounds and methods of using the compounds.

Description

Technical Field

[0001] The present invention relates to organic compounds useful for treatment and / or prevention in mammals, and particularly relates to the inhibition of KRAS G12C useful for treating cancer.

[0002] RAS is one of the most well-known cancer proto-genes. Approximately 30% of human cancers contain mutations in three of the most notable members, KRAS, HRAS, and NRAS, making them the most common oncogenic drivers. Generally, KRAS mutations are associated with poor prognosis, particularly in colorectal cancer, pancreatic cancer, and lung cancer. As the most frequently mutated RAS isoform, KRAS has been intensively studied in the past few years. Among the most commonly occurring 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 across colorectal cancer (CRC), pancreatic ductal adenocarcinoma (PDAC), and lung adenocarcinoma (LUAD). Notably, KRAS wild-type amplification is found in approximately 7% of all KRAS-altered cancers (ovary, esophagogastric, uterus) and is ranked among the top mutations.

[0003] All RAS proteins belong to the protein family of small GTPases that hydrolyze GTP to GDP. Structurally, KRAS is divided into an effector-binding lobe, followed by an allosteric lobe, and a carboxy-terminal region responsible for membrane anchoring. The effector lobe contains the P-loop, switch I, and switch II regions. The switch I / II loop plays an important role in KRAS downstream 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.

[0004] The KRAS protein switches from an inactive form to an active form via binding to GTP and GDP, respectively. Under physiological conditions, the transition between these two states is regulated by guanine nucleotide exchange factors (GEFs) such as Son Of Sevenless Homolog 1 (SOS1), which catalyze the exchange of GDP for GTP, or GTPase-activating proteins (GAPs) that enhance endogenous GTPase activity or accelerate RAS-mediated GTP hydrolysis. Inactive RAS-GDP is converted to active RAS-GTP that directly binds to the RAF RAS-binding domain (RAF RBD ), recruits the RAF kinase family from the cytoplasm to the membrane, where it dimerizes and becomes active. Activated RAF then performs a series of phosphorylation reactions on its downstream mitogen-activated protein kinases (MEK) and extracellular signal-regulated kinases (ERK) to propagate the growth signal. Among the RAF family of protein kinases (three known isoforms ARAF, BRAF, CRAF / RAF1), BRAF is the most frequently mutated and remains the most potent activator of MEK. Despite the different binding preferences shown by individual RAS and RAF family members, all RAFs have a conserved RBD for the forward transmission of MAPK signaling, which is frequently used to characterize KRAS inhibition (e.g., KRAS-BRAF in this specification RBD ). For KRAS, mutations at positions 12, 13, 61, and 146 result in a shift to the active KRAS form by impairing nucleotide hydrolysis or activating nucleotide exchange, leading to overactivation of the MAPK pathway that results in tumorigenesis.

[0005] Despite its well-recognized importance in cancer malignancy, past continuous efforts have not been able to develop an approved therapy for KRAS mutant cancers until recently, and the first-in-class drug AMG510 has been rapidly approved as a second-line treatment in KRAS G12C-driven non-small cell lung cancer (NSCLC). Nevertheless, clinically acquired resistance to KRAS G12C inhibitors strictly emerges with disease progression after approximately 6 months of treatment. Secondary RAS mutants in oncogenic hotspots (e.g., G12 / G13 / Q61) and within the switch II pocket (e.g., H95, R68, and Y96) have been observed where all mutations converge to reactivate RAS-MAPK signaling. Furthermore, over 85% of all KRAS mutant or wild-type amplified driver cancers still lack novel agents. In summary, both numerous escape mechanisms and diverse oncogenic alleles highlight the urgent medical need for further KRAS therapies. Therefore, the inventors have invented an oral compound that targets and inhibits KRAS alleles for the treatment of KRAS mutant-driven cancers.

[0006] First-generation KRAS G12C inhibitors such as sotorasib and adagrasib, which target the "GDP-bound off-form" (RAS OFF) of KRAS G12C, have demonstrated promising efficacy. This therapy has benefited many patients with activated KRAS gene mutations, but almost all patients who initially benefited ultimately acquire resistance through various mechanisms. Increases in cases of second mutations of KRAS G12C such as Y96D, R68S, H95D, H95Q, H95R, V8L have been confirmed from patient samples (Tanaka et al., Cancer Discovery (2021), Awad et al., NEJM (2021), Ho et al., EJC (2021), Zhao et al., Nature (2021), Tsai et al., JCI (2022)), or discovered from saturation mutagenesis (Siyu et al, PNAS (2022)) and ENU mutagenesis (Takamasa et al, J Thorac Oncol (2021)) that showed resistance to KRAS(OFF)G12C inhibitors. Therefore, there is an unmet need to prevent the acquisition of one or more mutations in RAS that confer resistance to RAS(OFF) inhibitors. SUMMARY OF THE INVENTION

[0007] The present invention relates to a novel compound of formula (I) TIFF2025522629000002.tif77170[wherein, R 1 is TIFF2025522629000003.tif35170; {In the above formula, R 8 is C 1-6 alkyl, R 9 is C 3-7 cycloalkyl, azetidinyl or phenyl, and the C 3-7 cycloalkyl, azetidinyl and phenyl are halo C 3-6 alkynyl, (halo C 3-6(Alkylpyrimidinyl)C 2-6 Alkynyl or pyrimidinyl C 2-6 Substituted with alkynyl} R 2 is C 1-6 alkyl; R 3 is H or halogen; R 4 is H or halogen; R 5 is C 1-6 alkyl or halo C 1-6 alkyl; R 6 is C 1-6 alkoxy C 1-6 alkyl; R 7 is morpholinyl, (halo C 1-6 alkyl)piperazinyl or C 1-6 alkylpiperazinyl; A 1 is thiazolylene; A 2 is C 1-6 alkylene; Provided that R 3 and R 4 are not both H at the same time] or a pharmaceutically acceptable salt thereof.

[0008] The present invention also relates to their production, medicaments based on the compounds according to the invention and their production, and the use of the compounds of formula (I) or (Ia) thereof as inhibitors of KRAS.

[0009] The compounds of the present invention address the issue of GSH toxicity compared to reference compounds. The compounds of formula (I) or (Ia) exhibit good KRAS inhibition against G12C, G12D, and G12V. In another embodiment, the compounds of the present invention demonstrated excellent cancer cell inhibition and human stem cell stability. Further, the compounds of formula (I) or (Ia) also exhibit good or improved cytotoxicity, solubility, and single-dose pharmacokinetic (SDPK) profiles. Additionally, the compounds of the present invention have demonstrated good efficacy against a second mutation as referred to in this application.

Brief Description of the Drawings

[0010]

Figure 1

Modes for Carrying Out the Invention

[0011] Definitions The term "C 1-6 alkyl" means a saturated straight-chain or branched-chain 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. Specific "C 1-6 alkyl" groups are methyl, ethyl, and n-propyl.

[0012] The term "C 1-6 alkoxy" means C 1-6 alkyl-O-.

[0013] The term "C 1-6 alkylene" means a straight-chain or branched saturated divalent hydrocarbon group of 1 to 6 carbon atoms or a bicyclic saturated divalent hydrocarbon group of 3 to 6 carbon atoms. Examples of C 1-6 alkylene groups include methylene, ethylene, propylene, 2-methylpropylene, butylene, 2-ethylbutylene, pentylene, hexylene.

[0014] The terms "halogen" and "halo" are used interchangeably herein and refer to fluoro, chloro, bromo, or iodo.

[0015] "C 2~6 The term "alkynyl" refers to a monovalent straight-chain or branched hydrocarbon group of 2 to 6 carbon atoms having at least one triple bond. In certain embodiments, alkynyl has 2 to 4 carbon atoms having at least one triple bond. C 2-6 Examples of alkynyl include ethynyl (-C≡CH), prop-1-ynyl (-C≡CCH3), prop-2-ynyl (propargyl, -CH2C≡CH), but-1-ynyl, but-2-ynyl, and but-3-ynyl.

[0016] The terms "halogen" and "halo" are used interchangeably herein and mean fluoro, chloro, bromo, or iodo.

[0017] "C 3-6 The term "alkynyl" means a monovalent straight-chain or branched hydrocarbon group of 3 to 6 carbon atoms having at least one triple bond. In certain embodiments, alkynyl has 3 to 4 carbon atoms having at least one triple bond. C 3-6 Examples of alkynyl include prop-1-ynyl (-C≡CCH3), prop-2-ynyl (propargyl, -CH2C≡CH), but-1-ynyl, but-2-ynyl, and but-3-ynyl.

[0018] "HaloC 1-6 The term "alkyl" means a C 1-6 alkyl group in which at least one of the hydrogen atoms of the alkyl group is replaced by the same or different halogen atoms, particularly fluorine atoms. Examples of haloalkyl include mono-fluoro-, di-fluoro-, or tri-fluoro-methyl, -ethyl, or -propyl, such as 3,3,3-trifluoropropyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, fluoromethyl, or trifluoromethyl. 1-6 ​

[0019] "HaloC 3-6 alkynyl" means a C 3-6 alkynyl group in which at least one of the hydrogen atoms of the alkynyl group is replaced by the same or different halogen atoms. HaloC 3-6 alkynyl. Examples of haloC 3-6 alkynyl include 3,3,3-trifluoroprop-1-ynyl.

[0020] "C 3-7 cycloalkyl" means a monovalent saturated monocyclic or bicyclic hydrocarbon group having 3 to 7 ring carbon atoms. Bicyclic means consisting of two saturated carbon rings having one or more carbon atoms in common. Examples of monocyclic cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. Examples of bicyclic cycloalkyl are bicyclo[1.1.0]butyl, bicyclo[2.2.1]heptanyl, bicyclo[1.1.1]pentanyl or bicyclo[2.2.2]octanyl.

[0021] The term "thiazolylene" means a divalent thiazolyl group.

[0022] The term "oxo" means a divalent oxygen atom =O.

[0023] The term "dimethylmethylene" means TIFF2025522629000004.tif14170.

[0024] The term "protecting group" means a group that selectively blocks a reaction site in a polyfunctional compound so that a chemical reaction is selectively carried out at another unprotected reaction site in the context generally relevant in the field of synthetic chemistry. The protecting group can be removed at an appropriate point. Exemplary protecting groups are amino protecting groups, carboxy protecting groups or hydroxy protecting groups.

[0025] One of ordinary skill in the art will understand that the following structures of the compounds of formula (Ia) and (Ia') are identical, particularly with respect to the chiral centers. TIFF2025522629000005.tif64170 The term "pharmaceutically acceptable salt" means a salt that is not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid addition salts and base addition salts.

[0026] The term "pharmaceutically acceptable acid addition salt" refers to salts formed from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from the classes of aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic acids, for example, 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, embonic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid and salicylic acid, which are pharmaceutically acceptable.

[0027] The term "pharmaceutically acceptable basic addition salt" refers to a pharmaceutically acceptable salt formed with an organic or inorganic base. Examples of acceptable inorganic bases include sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, and aluminum salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include primary, secondary, and tertiary amines, substituted amines including natural substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, and salts of polyamine resins.

[0028] The term "pharmaceutically active metabolite" means a pharmacologically active product produced through the metabolism in vivo of a specific compound or its salt. After entering the body, most drugs serve as substrates for chemical reactions and may change their physical properties and biological effects. These metabolic conversions usually affect the polarity of the compounds of the present invention and change the way drugs are distributed in the body and excreted from the body. However, in some cases, drug metabolism is required for therapeutic effects.

[0029] The term "therapeutically effective amount" means an amount of a compound or molecule of the invention that, when administered to a subject, (i) treats or prevents a particular disease, condition or disorder described herein, (ii) reduces, ameliorates or eliminates one or more symptoms of a particular disease, condition or disorder, or (iii) prevents or delays the occurrence of one or more symptoms of a particular disease, condition or disorder. The therapeutically effective amount will vary depending on the compound, the medical condition 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.

[0030] The term "pharmaceutical composition" means a mixture or solution containing a therapeutically effective amount of an active ingredient together with pharmaceutically acceptable additives, which is administered to a mammal in need thereof, such as a human.

[0031] The terms "pharmaceutically acceptable additive", "pharmaceutically acceptable carrier", and "therapeutically inert additive" are used interchangeably and mean any pharmaceutically acceptable component in a pharmaceutical composition that has no therapeutic activity and is non-toxic to the subject to which it is administered, for example, disintegrants, binders, fillers, solvents, buffers, tonicity agents, stabilizers, antioxidants, surfactants, carriers, diluents or lubricants used in formulating pharmaceutical products.

[0032] Inhibitor of KRAS The present invention relates to (i') a compound of formula (I) TIFF2025522629000006.tif75170[In the above formula, R 1 is TIFF2025522629000007.tif33170; {In the above formula, R 8 is C 1-6 alkyl, R 9 is C 3-7 cycloalkyl, azetidinyl or phenyl, said C 3-7Cycloalkyl, azetidinyl and phenyl are halo C 3-6 alkynyl, (halo C 3-6 alkylpyrimidinyl)C 2-6 alkynyl or pyrimidinyl C 2-6 substituted with alkynyl}{ R 2 is C 1-6 alkyl; R 3 is H or halogen; R 4 is H or halogen; R 5 is C 1-6 alkyl or halo C 1-6 alkyl; R 6 is C 1-6 alkoxy C 1-6 alkyl; R 7 is morpholinyl, (halo C 1-6 alkyl)piperazinyl or C 1-6 alkylpiperazinyl; A 1 is thiazolylene; A 2 is C 1-6 alkylene; However, R 3 and R 4 are not H at the same time] or a pharmaceutically acceptable salt thereof.

[0033] Another embodiment of the present invention is the compound of formula (Ia) (ii’) TIFF2025522629000008.tif75170[In the above formula, R 1 is TIFF2025522629000009.tif33170; {In the above formula, R 8 is C 1-6 alkyl, R 9 is C 3-7 cycloalkyl, azetidinyl or phenyl, and the C3-7 Cycloalkyl, azetidinyl and phenyl are halo C 3-6 Alkynyl, (halo C 3-6 Alkylpyrimidinyl)C 2-6 Alkynyl or pyrimidinyl C 2-6 Substituted with alkynyl} R 2 is C 1~6 alkyl; R 3 is H or halogen; R 4 is H or halogen; R 5 is C 1-6 alkyl or halo C 1-6 alkyl; R 6 is C 1-6 alkoxy C 1-6 alkyl; R 7 is morpholinyl, (halo C 1-6 alkyl)piperazinyl or C 1-6 alkylpiperazinyl; A 1 is thiazolylene; A 2 is C 1-6 alkylene; Provided that R 3 and R 4 are not both H at the same time] or a pharmaceutically acceptable salt thereof.

[0034] The present invention relates to (i) a compound of formula (I) TIFF2025522629000010.tif75170[In the above formula, R 1 is TIFF2025522629000011.tif29170; {In the above formula, R 8 is C 1-6 alkyl, R 9 is C 3-7 cycloalkyl, azetidinyl or phenyl, said C3-7 Cycloalkyl, azetidinyl and phenyl are halo C 3-6 Alkynyl or pyrimidinyl C 2-6 Substituted with alkynyl} R 2 is C 1-6 alkyl; R 3 is H or halogen; R 4 is H or halogen; R 5 is C 1-6 alkyl or halo C 1-6 alkyl; R 6 is C 1-6 alkoxy C 1-6 alkyl; R 7 is morpholinyl, (halo C 1-6 alkyl) piperazinyl or C 1-6 alkyl piperazinyl; A 1 is thiazolylene; A 2 is C 1-6 alkylene; Provided that R 3 and R 4 are not both H at the same time] or a pharmaceutically acceptable salt thereof.

[0035] Another embodiment of the present invention is (ii) a compound of formula (Ia) TIFF2025522629000012.tif75170[In the above formula, R 1 is TIFF2025522629000013.tif29170; {In the above formula, R 8 is C 1-6 alkyl, R 9 is C 3-7 cycloalkyl, azetidinyl or phenyl, and the C 3-7 cycloalkyl, azetidinyl and phenyl are halo C3-6 Alkynyl or pyrimidinyl C 2-6 substituted with alkynyl} R 2 is C 1-6 alkyl; R 3 is H or halogen; R 4 is H or halogen; R 5 is C 1-6 alkyl or halo C 1-6 alkyl; R 6 is C 1-6 alkoxy C 1-6 alkyl; R 7 is morpholinyl, (halo C 1-6 alkyl)piperazinyl or C 1-6 alkylpiperazinyl; A 1 is thiazolylene; A 2 is C 1-6 alkylene; provided that R 3 and R 4 are not both H at the same time] or a pharmaceutically acceptable salt thereof.

[0036] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in (iii)(i), (ii), (i') or (ii') or a pharmaceutically acceptable salt thereof, wherein R 1 is TIFF2025522629000014.tif29170, where R 8 is halo C 3-6 alkyl-substituted C 3-7 cycloalkyl.

[0037] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (iv)(i) to (iii), (i') and (ii') or a pharmaceutically acceptable salt thereof, wherein R 1 is TIFF2025522629000015.tif is 29170, where R 8 is methyl and R 9 is cyclobutyl substituted with 3,3,3-trifluoroprop-1-ynyl.

[0038] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (v) (i) to (iv), (i') and (ii'), wherein R 9 is 3-(3,3,3-trifluoroprop-1-ynyl)cyclobutyl.

[0039] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (vi) (i) to (v), (i') and (ii') or a pharmaceutically acceptable salt thereof, wherein R 2 is isopropyl.

[0040] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (vii) (i) to (vi), (i') and (ii') or a pharmaceutically acceptable salt thereof, wherein R 3 is halogen.

[0041] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (viii) (i) to (vii), (i') and (ii') or a pharmaceutically acceptable salt thereof, wherein R 3 is fluoro.

[0042] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (ix) (i) to (viii), (i') and (ii') or a pharmaceutically acceptable salt thereof, wherein R 4 is H or fluoro.

[0043] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (x)(i)-(ix), (i') and (ii'), or a pharmaceutically acceptable salt thereof, wherein R 4 is H.

[0044] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (xi)(i)-(x), (i') and (ii'), or a pharmaceutically acceptable salt thereof, wherein R 5 is ethyl or 2,2,2-trifluoroethyl.

[0045] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (xii)(i)-(xi), (i') and (ii'), or a pharmaceutically acceptable salt thereof, wherein R 6 is 1-methoxyethyl.

[0046] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (xiii)(i)-(xii), (i') and (ii'), or a pharmaceutically acceptable salt thereof, wherein R 7 is morpholinyl, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or 4-methylpiperazin-1-yl.

[0047] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (xiv)(i)-(xiii), (i') and (ii'), or a pharmaceutically acceptable salt thereof, wherein A 1 is TIFF2025522629000016.tif22170, where the bond "a" is attached to the indole ring.

[0048] A further embodiment of the present invention is a compound of formula (I) or (Ia) as described in any one of (xv)(i)-(xiv), (i') and (ii'), or a pharmaceutically acceptable salt thereof, wherein A 2 is dimethylmethylene.

[0049] Another embodiment of the present invention is a compound of formula (I) or (Ia) as described in (xvi) (i) or (ii), (i') or (ii') [wherein, R 1 is TIFF2025522629000017.tif29170 (wherein R 8 is C1-6 alkyl; R 9 is halo C 3-6 cycloalkyl substituted with alkynyl); 3-7 R 2 is C 1-6 alkyl; R 3 is halogen; R 4 is H; R 5 is C 1-6 alkyl or halo C 1-6 alkyl; R 6 is C 1-6 alkoxy C 1-6 alkyl; R 7 is morpholinyl, (halo C 1-6 alkyl) piperazinyl or C 1-6 alkyl piperazinyl; A 1 is TIFF2025522629000018.tif22170 (wherein the bond "a" is attached to the indole ring); A 2 is C 1-6 alkylene] or a pharmaceutically acceptable salt thereof.

[0050] A further embodiment of the present invention is a compound of formula (I) or formula (Ia) as described in (xvii) (xvi) [wherein, R 1 is TIFF2025522629000019.tif29170 (wherein R 8 ​is methyl; R 9 is 3-(3,3,3-trifluoroprop-1-ynyl)cyclobutyl; R 2 is isopropyl; R 3 is fluoro; R 4 is H; R 5 is ethyl or 2,2,2-trifluoroethyl; R 6 is (1S)-1-methoxyethyl; R 7 is morpholinyl, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or 4-methylpiperazin-1-yl; A 1 is TIFF2025522629000020.tif22170 (wherein the bond "a" is bonded to the indole ring); A 2 is dimethylmethylene] or a pharmaceutically acceptable salt thereof.

[0051] Another embodiment of the present invention is a compound of formula (I) or (Ia) selected from (xviii) the following: trans-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide; N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide; N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)benzamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(2-pyrimidin-2-ylethynyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide; trans-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[5-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutanecarboxamide; N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide; N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.12,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[4-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutanecarboxamide; N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide; (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-isopropyl-4-oxo-4-[3-(3,3,3-trifluoroprop-1-ynyl)azetidin-1-yl]butanamide; N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.12,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[4-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutanecarboxamide; and cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 [Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide or a pharmaceutically acceptable salt thereof.

[0052] Another embodiment of the present invention is (xix) the following steps: a) In the presence of a coupling reagent and a base, a compound of formula (II) TIFF2025522629000021.tif64170 and an acid (III) A step of subjecting TIFF2025522629000022.tif25170 to a coupling reaction to form a compound of formula (I), wherein In the above formula, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , A 1 and A 2 are defined as being in any one of (i) to (xvii); the coupling reagent is T3P, HATU, PyBOP or EDCI / HOBt; and the base is TEA, DIEPA or DMAP, the step relates to a process for preparing a compound according to any one of (i) to (xvii) comprising

[0053] Another embodiment of the present invention is a compound or a pharmaceutically acceptable salt according to any one of (i) to (xviii), (i') and (ii') for use as a (xx) therapeutic active substance.

[0054] Another embodiment of the present invention is a pharmaceutical composition comprising a compound according to any one of (i) to (xviii), (i') and (ii') and a pharmaceutically acceptable additive (xxi).

[0055] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for treating a (xxii) KRAS G12C protein-related disease.

[0056] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii) for treating a (xxiii) KRAS G12C, G12D and G12V protein-related disease.

[0057] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for inhibiting the RAS interaction with downstream effectors, wherein the downstream effectors are RAF and PI3K.

[0058] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for inhibiting proliferative oncogenic MAPK and PI3K signaling.

[0059] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for the treatment or prevention of KRAS mutant-driven cancer, wherein the cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma, ovarian cancer and endometrial cancer.

[0060] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for the treatment or prevention of KRAS mutant-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0061] Another embodiment of the present invention is a compound or a pharmaceutically acceptable salt thereof according to any one of (i) to (xviii), (i') and (ii') for the treatment or prevention of KRAS mutant-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0062] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for the treatment or prevention of KRAS mutant-driven cancer, wherein the cancer comprises a first mutation that is G12C and a second mutation at a position selected from V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L.

[0063] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for the preparation of a medicament for the treatment or prevention of KRAS mutant-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0064] Another embodiment of the present invention is the use of a compound according to any one of (i) to (xviii), (i') and (ii') for the preparation of a medicament for the treatment or prevention of KRAS mutant-driven cancer, wherein the cancer comprises a first mutation that is G12C and a second mutation at a position selected from V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L.

[0065] Another embodiment of the present invention is a method for the treatment or prevention of KRAS mutant-driven cancer, which comprises administering a therapeutically effective amount of a compound as defined in any one of (i) to (xviii), (i') and (ii'), wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0066] Another embodiment of the present invention is a method for the treatment or prevention of KRAS mutant-driven cancer, wherein the cancer comprises a first mutation that is G12C and a second mutation at a position selected from V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L.

[0067] Another embodiment of the present invention is a compound or a pharmaceutically acceptable salt according to any one of (i)-(xviii), (i') and (ii'), when manufactured according to the method described in (xxxiv)(xix).

[0068] Pharmaceutical Compositions and Administration In another embodiment, there is provided a pharmaceutical composition or medicament containing a compound of the present invention and a therapeutically inert carrier, diluent or additive, and a method of using a compound of the present invention for preparing such composition and medicament. In one example, a compound of formula I can be formulated into a galenical dosage form by mixing, at ambient temperature, with a physiologically acceptable carrier, i.e., a carrier that is non-toxic to the recipient at the dosage and concentration used, at an appropriate pH and with a desired purity. The pH of the formulation depends mainly on the particular use and the concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, a compound of formula (I) is formulated in an acetate buffer at pH 5. In another embodiment, a compound of formula (I) is sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0069] The composition is formulated, administered, and managed in a manner consistent with medical practice guidelines. Factors to be considered in this context include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the dosing schedule, and other factors known to the medical practitioner. The "effective amount" of the compound to be administered is determined by such considerations and is the minimum amount necessary to inhibit the interaction between mutant RAS (e.g., KRAS G12C) and RAF and block oncogenic MAPK signaling. For example, such an amount may be less than an amount that is toxic to normal cells or the entire mammal.

[0070] In one example, the pharmaceutically effective amount of the compound of the invention administered parenterally per dose is in the range of about 0.1 to 1000 mg / kg, or about 0.1 to 1000 mg per kg of patient body weight per day, and a typical initial range of the compound used is 0.3 to 15 mg / kg / day. In another embodiment, an oral unit dosage form such as a tablet or capsule preferably contains about 1 to about 1000 mg of the compound of the invention.

[0071] The compounds of the present invention can be administered by any suitable means including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intralung, intradermal, intrathecal, epidural, and intranasal, and, if desired in local treatment, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

[0072] The compounds of the present invention can be administered in any convenient dosage form such as tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions can include ingredients conventional in pharmaceutical preparations such as diluents, carriers, pH adjusters, sweeteners, bulking agents, and additional active agents.

[0073] Typical formulations are prepared by mixing the compounds of the present invention with carriers or additives. Suitable carriers and additives 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 formulations may also include one or more buffers, stabilizers, surfactants, wetting agents, smoothing agents, emulsifiers, suspending agents, preservatives, antioxidants, opacifying agents, flow promoters, processing aids, colorants, sweeteners, fragrances, flavoring agents, diluents, and other known additives for presenting the drug (i.e., the compound of the present invention or its pharmaceutical composition) well or for assisting in the manufacture of a pharmaceutical product (i.e., a medicine).

[0074] Examples of suitable oral dosage forms include tablets containing about 1 to 1000 mg of the compound of the present invention, 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 powder components are first mixed together and then mixed with a solution of PVP. 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, for example, 5 to 400 mg of the compound of the present invention in a suitable buffer solution, such as phosphate buffer, and adding an isotonic agent (such as a salt like sodium chloride) if desired. The solution may be filtered using, for example, a 0.2 micron filter to remove impurities and contaminants.

[0075] Accordingly, certain embodiments include a pharmaceutical composition comprising a compound of formula I or a stereoisomer or pharmaceutically acceptable salt thereof. In further embodiments, there is provided a pharmaceutical composition comprising a compound of formula I or a stereoisomer or pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier or additive.

[0076] Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in the treatment of mutant KRAS-driven cancer. Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in the treatment of mutant KRAS-driven cancer.

[0077] The following Compositions A and B illustrate typical compositions of the present invention and are presented merely as representatives thereof.

[0078] Composition A The compound of the present invention can be used in a method known per se as an active ingredient for producing tablets of the following composition. TIFF2025522629000023.tif48170Composition B The compounds of the present invention can be used in a manner known per se as active ingredients for the production of capsules of the following compositions. TIFF2025522629000024.tif55170

[0079] Indications and Therapeutic Methods By promoting the formation of a high-affinity ternary complex between the compounds of the present invention and cyclophilin A (CYPA) which is widely expressed with the KRAS protein, the compounds of the present invention induce a new binding pocket in KRAS, which inhibits the KRAS interaction with downstream effectors such as RAF and PI3K. Therefore, the compounds of the present invention are useful for inhibiting the proliferative oncogenic MAPK and PI3K signal transduction and reducing cell proliferation, particularly cancer cells. The compounds of the present invention are useful for terminating RAS signal transduction in cells expressing RAS variants, such as KRAS mutation-driven pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma ovarian cancer, endometrial cancer, etc. Alternatively, the compounds of the present invention are useful for terminating RAS signal transduction in malignant solid tumors in which the oncogenic role of KRAS mutations is enhanced by dysregulation or mutation of effector pathways such as MAPK, PI3K-AKT-mTOR (mammalian target of rapamycin) driven signal transduction, and for targeted therapy of pancreatic adenocarcinoma, colorectal cancer, non-small cell lung cancer, etc.

[0080] Another embodiment includes a method for treating or preventing cancer in a mammal in need thereof, the method comprising administering to the mammal a therapeutically effective amount of a compound of formula (I), a stereoisomer, a tautomer, a prodrug or a pharmaceutically acceptable salt thereof.

[0081] Synthesis The compounds of the present invention can be prepared by any conventional means. Suitable methods for synthesizing these compounds and their starting materials are shown in the following schemes and examples. All substituents, especially R 1 ~R 7 and A 1 ~A 2Unless otherwise indicated, it is as defined above. Further, unless otherwise specified, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to those skilled in the art of organic chemistry.

[0082] A general synthetic route for preparing the compound of formula (I) is shown below.

[0083] Scheme 1 The compound of formula II was synthesized according to the procedure described for intermediates A to K in TIFF2025522629000025.tif53170. The compound of formula (I) can be obtained by a coupling reaction between acid (III) and the compound of formula (II) using a coupling reagent such as T3P, HATU, PyBOP and EDCI / HOBt in the presence of a base such as TEA, DIEPA and DMAP.

[0084] The compounds of the present invention can be obtained as mixtures of diastereomers or enantiomers which can be separated by methods well known in the art, for example, by (chiral) HPLC or SFC. In another embodiment, the compounds of formula (I) can be obtained according to the above scheme by using the corresponding chiral starting materials.

[0085] The present invention also relates to a method for preparing a compound of formula (I) comprising the following steps: a) reacting a compound of formula (II) TIFF2025522629000026.tif64170 with an acid (III) TIFF2025522629000027.tif25170 in a coupling reaction in the presence of a coupling reagent and a base to form a compound of formula (I), wherein in step a), the coupling reagent can be, for example, T3P, HATU, PyBOP or EDCI / HOBt; and the base can be, for example, TEA, DIEPA or DMAP.

[0086] Compounds of formula (I) or (Ia) when produced according to the above method are also an object of the present invention.

Examples

[0087] The present invention will be more fully understood by reference to the following examples. However, they are not to be construed as limiting the scope of the present invention.

[0088] Abbreviations The present invention will be more fully understood by reference to the following examples. However, they are not to be construed as limiting the scope of the present invention.

[0089] The abbreviations used in this specification are as follows. TIFF2025522629000028.tif252170TIFF2025522629000029.tif150170

[0090] General experimental conditions Intermediates and final compounds were purified by flash chromatography using one of the following apparatuses: i) Biotage SP1 system and Quad 12 / 25 Cartridge module, ii) ISCO Combiflash chromatography apparatus. Silica gel brand and pore size: i) KP-SIL 60Å, particle size: 40 - 60μm; ii) CAS registration number: silica gel: 63231-67-4, particle size: 47 - 60 microns of silica gel; iii) ZCX of Qingdao Ocean Chemical Co., Ltd., pore size: 200 - 300 or 300 - 400.

[0091] The intermediates and final compounds were purified by preparative HPLC on a reversed-phase column using an XBridge™ Prep-C18 (5 μm, OBD™ 30×100 mm) column, a SunFire™ Prep-C18 (5 μm, OBD™ 30×100 mm) column, a Phenomenex Synergi-C18 (10 μm, 25×150 mm) or a Phenomenex Gemini-C18 (10 μm, 25×150 mm). Waters AutoP purification system (sample manager 2767, pump 2525, detector: Micromass ZQ and UV 2487, solvent system: acetonitrile and 0.1% aqueous ammonium hydroxide solution; acetonitrile and 0.1% FA aqueous solution or acetonitrile and 0.1% TFA aqueous solution). Or, a Gilson-2811 purification system (pump 322, detector: UV 156, solvent system: acetonitrile and 0.05% aqueous ammonium hydroxide solution; acetonitrile and 0.225% FA aqueous solution; acetonitrile and 0.05% HCl aqueous solution; acetonitrile and 0.075% TFA aqueous solution; or acetonitrile and water).

[0092] For SFC chiral separation, the intermediate was separated on a chiral column (Daicel chiralpak IC, 5 μm, 30×250 mm), AS (10 μm, 30×250 mm) or AD (10 μm, 30×250 mm) using a Mettler Toledo Multigram III system SFC, Waters 80Q preparative SFC or Thar 80 preparative SFC, solvent system: CO2 and IPA (0.5% TEA in IPA) or CO2 and MeOH (0.1% NH3·H2O in MeOH), back pressure 100 bar, detection UV@254 or 220 nm.

[0093] The LC / MS spectrum of the compound was obtained using LC / MS (Waters™ Alliance 2795 - Micromass ZQ, Shimadzu Alliance 2020 - Micromass ZQ or Agilent Alliance 6110 - Micromass ZQ). The LC / MS conditions were as follows (runtime 3 minutes or 1.5 minutes): Acidic condition I: A: 0.1% TFA in H2O; B: 0.1% TFA in acetonitrile; Acidic condition II: A: 0.0375% TFA in H2O; B: 0.01875% TFA in acetonitrile; Basic condition I: A: 0.1% NH3·H2O in H2O; B: acetonitrile; Basic condition II: A: 0.025% NH3·H2O in H2O; B: acetonitrile; Neutral condition: A: H2O; B: acetonitrile.

[0094] Mass spectra (MS): Generally, only the ions indicating the parent mass are reported, and unless otherwise stated, the cited mass ions are positive mass ions (MH) + is.

[0095] The NMR spectrum was obtained using a Bruker Avance 400 MHz.

[0096] The microwave-assisted reactions were carried out using a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were conducted under an argon or nitrogen atmosphere. The reagents were used as received from commercial suppliers without further purification, unless otherwise stated.

[0097] Preparative Examples Preparation of Intermediates Intermediate A 1 - [6 - [(1S) - 1 - methoxyethyl - 5 - (4,4,5,5 - tetramethyl - 1,3,2 - dioxaborolan - 2 - yl) - 3 - pyridyl] - 4 - methyl - piperazine TIFF2025522629000030.tif Intermediate A in the title was prepared according to the following scheme. TIFF2025522629000031.tif112170

[0098] Step 1: Preparation of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (Compound A2) To a solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (Compound A1, 2.0 g, 9.26 mmol) and bis(pinacolato)diboron (3.5 g, 13.9 mmol) in THF (30 mL) were added 4,4'-di-tert-butyl-2,2'-bipyridine (372.7 mg, 1.39 mmol) and [Ir(OMe)(COD)]2 (306.3 mg, 0.460 mmol). The mixture was stirred at 75 °C for 16 h under N2 protection. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (EA / PE: 0 - 20%) to give 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (Compound A2, 2.4 g) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ ppm 8.91 (d, J = 1.4 Hz, 1 H), 8.21 (d, J = 1.4 Hz, 1 H), 4.95 (q, J = 6.5 Hz, 1 H), 3.30 (s, 3 H), 1.49 (d, J = 6.5 Hz, 3 H), 1.35 (s, 12 H).

[0099] Step 2: Preparation of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3) To a solution of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (Compound A2, 2.5 g, 7.3 mmol) in ACN (40 mL) was added N-iodosuccinimide (4.1 g, 18.27 mmol). The mixture was stirred at 90 °C for 40 h under N2 protection. The reaction was quenched with a saturated solution of Na2SO3 (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 chromatography (EA / PE: 0 - 20%) to obtain 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 + ).

[0100] Step 3: Preparation of benzyl 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound A5) 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) were added cesium carbonate (1.6 g, 4.83 mmol), (R)-BINAP (60.1 mg, 0.1 mmol), and palladium(II) acetate (43.3 mg, 0.19 mmol). The mixture was stirred at 100 °C for 12 h under N2 protection. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel chromatography (EA / PE: 0 - 50%) to obtain 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 + ).

[0101] Step 4: Preparation of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]-4-methyl-piperazine (Intermediate A) To a solution of benzyl 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound A5, 740 mg, 1.7 mmol) and bis(pinacolato)diborane (519.2 mg, 2.04 mmol) in toluene (12 mL) were added KOAc (418.0 mg, 4.26 mmol) and Pd(dppf)Cl2 (124.7 mg, 0.170 mmol). The reaction mixture was stirred at 90 °C for 12 h under a N2 atmosphere. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by silica gel column to give 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]-4-methyl-piperazine (Intermediate A, 470 mg) as a brown solid. MS calculated value 482.3 (MH + ), measured value 482.2 (MH + ).

[0102] Intermediate B Methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylate Intermediate B was prepared according to the following scheme of TIFF2025522629000032.tif56170 or less. TIFF2025522629000033.tif134170

[0103] Step 1: Preparation of (4-bromothiazol-2-yl)methanol (Compound B2) To a solution of 4-bromothiazole-2-carboxaldehyde (Compound B1, 6.0 g, 31.25 mmol) in methanol (70 mL) was added sodium borohydride (1.7 g, 46.87 mmol) at 0 °C. The mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (300 mL) at 0 °C, and the reaction mixture was extracted with ethyl acetate (200 mL, 3 times). The combined organic phases were washed with brine (150 mL, 2 times), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain (4-bromothiazol-2-yl)methanol (Compound B2, 6 g) as a colorless oil.

[0104] Step 2: Preparation of 4-bromo-2-(bromomethyl)thiazole (Compound B3) To a solution of (4-bromothiazol-2-yl)methanol (Compound B2, 6.0 g, 30.92 mmol) in DCM (80 mL) were added CBr4 (15.4 g, 46.38 mmol) and triphenylphosphine (12.1 g, 46.38 mmol) at 0 °C. After stirring at 25 °C for 1 hour, the mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column eluting with ethyl acetate (0 - 10%) in petroleum ether to obtain (4-bromothiazol-2-yl)methyl bromide (Compound B3, 6.0 g) as a yellow oil. MS calculated value 255.9 (MH + ), measured value 255.9 (MH + ).

[0105] Step 3: Preparation of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5) (R)-2,5-Dihydro-3,6-dimethoxy-2-isopropylpyrazine (Compound B4, 4.3 g, 23.45 mmol) in a mixture of THF (60 mL) was slowly added with n-butyllithium (10 mL, 25.22 mmol, 2.5 M) at -78 °C. After the addition, the mixture was stirred at -78 °C for 0.5 h. 4-Bromo-2-(bromomethyl)thiazole (Compound B3, 5.4 g, 21.02 mmol) was added to the above mixture at -78 °C and further stirred for 1 h. The reaction was quenched with a saturated solution of NH4Cl (100 mL), and the reaction mixture was extracted with EtOAc (100 mL, 2 times). 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 reverse-phase chromatography to obtain 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5, 3.6 g) as a yellow oil. MS calculated value 360 (MH + ), measured value 359.9 (MH + ).

[0106] Step 4: Preparation of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (Compound B6) To a solution of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5, 3.6 g, 10 mmol) in ACN (20 mL) was added hydrochloric acid (66.62 mL, 0.3 M). The mixture was stirred at 25 °C for 2 h. The mixture was basified to pH = 8 with a saturated aqueous solution of NaHCO3. The mixture was extracted with EtOAc (80 mL, 6 times). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (Compound B6, 3.1 g) as a yellow oil. MS calculated value 264.9 (MH + ), measured value 264.9 (MH + ).

[0107] Step 5: Preparation of Methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoate (Compound B7) To a solution of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (B6, 3.1 g, 11.69 mmol) in DCM (40 mL) were added triethylamine (2.9 g, 29.23 mmol) and (Boc)2O (3.8 g, 17.54 mmol). After stirring at 30 °C for 12 h, the mixture was concentrated under vacuum. The residue was purified by silica gel column, eluting with ethyl acetate in petroleum ether (0 - 30%) to give methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoate (Compound B7, 3.2 g) as a yellow oil. MS calculated value 387 (MNa + ), measured value 386.9 (MNa + ).

[0108] Step 6: Preparation of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (Compound B8) To a solution of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoate (Compound B7, 3.2 g, 8.76 mmol) in THF (30 mL) / methanol (2 mL) / water (10 mL) was added lithium hydroxide (0.41 mL, 43.81 mmol). After stirring at 25 °C for 1 h, the reaction mixture was acidified with 1 M HCl until pH = 5. This mixture was extracted with EtOAc (40 mL, 2 times). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (Compound B8, 3.1 g) as a yellow oil. MS calculated value 373 (MNa + ), measured value 372.9 (MNa + ).

[0109] Step 7: Methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylate (Intermediate B) To a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (Compound B8, 3.1 g, 8.83 mmol) in DCM (50 mL) were added methyl (3S)-hexahydropyridazine-3-carboxylate; hydrochloride (Compound B9, 2.4 g, 13.24 mmol), EDCI (3.4 g, 17.65 mmol), 1-hydroxybenzotriazole (238.5 mg, 1.77 mmol) and NMM (9.92 mL, 88.26 mmol) at 0 °C. After stirring at 25 °C for 1 h, the reaction mixture was diluted with water (60 mL) and extracted with EtOAc (60 mL, 3 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, eluting with ethyl acetate in petroleum ether (10~30%) to give methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylate (Intermediate E, 2.4 g). MS calculated value 477 (MH + ), measured value 476.9 (MH + ).

[0110] 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione Intermediate C of the title was prepared according to the following scheme for TIFF2025522629000034.tif with 59170 below. TIFF2025522629000035.tif with 196170, TIFF2025522629000036.tif with 206170

[0111] Step 1: Preparation of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (Compound C3) A solution of SnCl4 (97.2 mL, 121.5 mmol) was slowly added to a mixture of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (Compound C1, 35.0 g, 116.8 mmol) in DCM (400 mL) at 0 °C. After stirring at -40 °C for 0.5 h, a solution of 5-bromo-6-fluoro-1H-indole (Compound C2, 25.0 g, 116.8 mmol) in DCM (200 mL) was added dropwise to this mixture, and it was stirred at -40 °C for 15 min. After completion of the reaction, it was quenched with saturated aqueous NaHCO3 (800 mL), and the reaction mixture was extracted with EtOAc (900 mL, 2 times). The combined organic layers were washed with brine (700 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was triturated with a solution (100 mL, petroleum ether:ethyl acetate = 8:1) and filtered. The filter cake was dried in vacuo to obtain 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (Compound C3, 50.0 g) as a yellow solid. MS calculated value 552.1 (MH + ) and measured value 552.1 (MH + ).

[0112] Step 2: Preparation of [3-(5-bromo-6-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C4) To a mixture of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (Compound C3, 50.0 g, 90.49 mmol) in THF (600 mL) was added dropwise LiBH4 (48.4 mL, 193.49 mmol, 4 M in THF) at 0 °C. The mixture was stirred at 70 °C for 24 h under a nitrogen atmosphere. After completion of the reaction, the reaction was quenched by slowly adding water (600 mL) at 0 °C, and the reaction mixture was extracted with EtOAc (600 mL, 2 times). The combined organic layers were washed with brine (600 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 20 - 33%) 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 + ).

[0113] 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) [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) were added with silver trifluoromethanesulfonate (20.3 g, 78.88 mmol) at 0 °C. The mixture was stirred at 0 °C for 10 minutes. After completion of the reaction, it was quenched with saturated aqueous 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 in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 0% - 2.5%) to give [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 43.0 g) as a yellow solid. MS calculated value 664.0 (MH + ), measured value 664.1 (MH + ).

[0114] Step 4: Preparation of Benzyl 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) [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 benzyl 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]piperazine-1-carboxylate (Intermediate A, 16.7 g, 34.69 mmol) were added to a mixed solution of 1,4-dioxane (270 mL) / toluene (90 mL) / water (90 mL), and potassium phosphate (15.7 g, 73.92 mmol) and Pd(dppf)Cl2 (920 mg, 1.26 mmol) were added thereto. This mixture was stirred at 70 °C for 12 hours under a nitrogen atmosphere. After completion of the reaction, the mixture was filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 20 - 50%) to obtain 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 + ).

[0115] Step 5: Preparation of benzyl 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-carboxylate (Compound C7) 4-[(5-Bromo-3-(3-(tert-butyl(diphenyl)silyl)oxy-2,2-dimethylpropyl)-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) were added dropwise with 2,2,2-trifluoroethyl trifluoromethanesulfonate (37.7 g, 162.56 mmol) at 0 °C, and the mixture was stirred at 20 °C for 12 hours. After completion of the reaction, EtOAc (70 mL) and water (100 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (70 mL, 2 times). The combined organic layers were washed with brine (100 mL, 4 times), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by silica gel column chromatography to give benzyl 4-[(5M)-5-[5-bromo-3-(3-(tert-butyl(diphenyl)silyl)oxy-2,2-dimethylpropyl)-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-((1S)-1-methoxyethyl)-3-pyridyl]piperazine-1-carboxylate (Compound C7, 8.0 g, peak 1, eluted first) as a yellow oil. MS calculated value 973.3 (MH + ), measured value 973.2 (MH + ).

[0116] Step 6: Preparation of benzyl 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-((1S)-1-methoxyethyl)-3-pyridyl]piperazine-1-carboxylate (Compound C8) Benzyl 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C7, 10.5 g, 10.78 mmol) in DMF (130 mL) was added cesium fluoride (8.2 g, 53.9 mmol), and the mixture was stirred at 60 °C for 24 h. After completion of the reaction, EtOAc (100 mL) and water (100 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (100 mL, 2 times). The combined organic layers were washed with brine (80 mL, 3 times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 25 - 66%) to give benzyl 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C8, 6.5 g) as a yellow solid. MS calculated value 735.2 (MH + ), measured value 735.1 (MH + ).

[0117] Step 7: Preparation of benzyl 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C9) Benzyl 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C8, 5.4 g), bis(pinacolato)diboron (2.8 g, 11.01 mmol) and potassium acetate (1.2 mL, 18.35 mmol) in toluene (70 mL) solution, Pd(dppf)Cl2 (537.1 mg, 0.73 mmol) was added. The mixture was degassed and purged three times with nitrogen atmosphere, and the mixture was stirred at 90 °C for 12 hours. After completion of the reaction, the mixture was cooled to room temperature. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 25 - 66%) to give benzyl 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C9, 5.2 g) as a yellow oil. MS calculated value 783.3 (MH + ), measured value 783.3 (MH + ).

[0118] Step 8: Preparation of methyl (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propanoyl]hexahydropyridazine-3-carboxylate (Compound C10) Methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylate (Intermediate B, 2.7 g, 5.69 mmol), benzyl 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C9, 4.9 g, 6.32 mmol) in toluene (60 mL) / 1,4-dioxane (20 mL) / water (20 mL) solution, under a nitrogen atmosphere, K3PO4 (3.4 g, 15.81 mmol) and Pd(dtbpf)Cl2 (412.2 mg, 0.63 mmol) were added. The mixture was stirred at 70 °C for 12 h. After completion of the reaction, the mixture was concentrated in vacuo to give a residue. The residue was purified by silica column (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-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propanoyl]hexahydropyridazine-3-carboxylate (Compound C10, 3.6 g) as a brown solid. MS calculated value 1053.4 (MH + ), measured value 1053.3 (MH + ).

[0119] Step 9: Preparation of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-Benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid (Compound C11) To a solution of methyl (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propanoyl]-hexahydropyridazine-3-carboxylate (Compound C10, 3.6 g, 3.42 mmol) in DCE (50 mL) was added trimethylstannanol (2.4 g, 13.67 mmol), and the mixture was stirred at 60 °C for 12 h. After completion of the reaction, EtOAc (80 mL) and water (60 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (80 mL, 2 times). 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-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid (Compound C11, 4.3 g) as a brown solid. MS calculated 1039.4 (MH + +), found 1039.2 (MH + +).

[0120] Step 10: Preparation of Benzyl 4-[5-[(7S,13S)-7-(tert-Butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C12) (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-Benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid (Compound C11, 4.3 g, 4.14 mmol) in DCM (430 mL) was added with DIEA (14.4 mL, 82.76 mmol), EDCI (11.9 g, 62.07 mmol) and 1-hydroxybenzotriazole (1.4 g, 10.35 mmol) at 0 °C. The mixture was stirred at 15 °C for 12 h. After completion of the reaction, the mixture was concentrated in vacuo, diluted with water (80 mL), and extracted with EtOAc (80 mL, 2 times). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 25% - 66%) to give Benzyl 4-[5-[(7S,13S)-7-(tert-Butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C12, 3.1 g) was obtained as a yellow gum. MS calculated value 1021.4 (MH + ), measured value 1021.2 (MH + ).

[0121] Step 11: Preparation of tert-butyl 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 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound C13) Benzyl 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (Compound C12, 3.1 g, 3.04 mmol) and aqueous formaldehyde solution (775.0 mg, 9.55 mmol) in methanol (150 mL) were mixed, and Pd(OH)2 (2.79 g, 3.97 mmol) on activated carbon was added. The mixture was degassed and purged three times with H2. The mixture was hydrogenated at 30 °C for 18 hours. After completion of the reaction, the mixture was filtered and the filtrate was concentrated in vacuo to give tert-butyl 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound C13, 2.6 g) was obtained as a brown solid. MS calculated value 901.3 (MH + ), measured value 901.3 (MH + ).

[0122] Step 12: (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C) tert-Butyl 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound C13, 2.6 g, 2.89 mmol) in DCM (18 mL) was added TFA (14.0 mL, 181.72 mmol). The mixture was stirred at 15 °C for 0.5 h. After completion of the reaction, the mixture was concentrated in vacuo, diluted with saturated NaHCO3 (30 mL), and extracted with EtOAc (30 mL, 3 times). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, concentrated in vacuo, and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C, 2.0 g) was obtained as a yellow solid and used as such in the next step. MS calculated value 801.3 (MH + ), measured value 801.2 (MH + ).

[0123] 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-tetrazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione TIFF2025522629000037.tif661702,2,2-Trifluoroethyl trifluoromethanesulfonate was replaced with iodoethane, and the title compound was prepared in the same manner as the preparation of Intermediate C.

[0124] 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione TIFF2025522629000038.tif59170This compound was prepared according to the following scheme. TIFF2025522629000039.tif194170TIFF2025522629000040.tif132170

[0125] Step 1: Preparation of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2) A mixture of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3, 2.03 g, 5.95 mmol) and 1-(2,2,2-trifluoroethyl)piperazine (Compound E1, 1.0 g, 5.95 mmol) in toluene (15 mL) was added with 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). The reaction mixture was degassed and purged with nitrogen three times, and the mixture was stirred at 100 °C for 12 h under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography to give 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2, 2.0 g) as a yellow oil. MS calculated value 382.2 (MH + ), measured value 382.1 (MH + ).

[0126] Step 2: 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3) 1-[5-Bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2, 3.2 g, 8.37 mmol), bis(pinacolato)diboron (3.19 g, 12.56 mmol) and KOAc (2.1 g, 20.93 mmol) in toluene (50 mL) were added Pd(dppf)Cl2 (306.3 mg, 0.42 mmol). The mixture was degassed and purged three times with nitrogen, and the mixture was stirred at 90 °C for 12 hours under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo to give a residue, which was purified by reverse-phase column to give 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3, 1.9 g) as a yellow gum. MS calculated value 430.2 (MH + ), measured value 348.4 (M-C6H 10 +H + ).

[0127] 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-dioxaborolan-2-yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3, 1.9 g, 4.41 mmol), [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 2.1 g, 3.15 mmol) in 1,4-dioxane (24 mL) / water (8 mL) / toluene (8 mL) were added K3PO4 (2.1 g, 9.5 mmol) and Pd(dppf)C l2(231 mg, 0.37 mmol) was added. The mixture was degassed by nitrogen bubbling for 2 minutes, and the reaction mixture was stirred at 70 °C for 12 hours. After cooling to room temperature, the reaction mixture was filtered. The filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (EtOAc in PE: 30% - 60%) to give [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E4, 960.0 mg) as a yellow gum. MS calculated value 839.3 (MH + ), measured value 839.3 (MH + ).

[0128] 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) [3-[5-Bromo-6-fluoro-(2M)-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) was added with Cs2CO3 (1.1 g, 3.44 mmol) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (2.7 g, 11.63 mmol) at 0 °C. After stirring at 20 °C for 15 h, the reaction mixture was poured into water (100 mL) and extracted with EtOAc (50 mL, 3 times). The combined organics were washed with brine (50 mL, 3 times), dried over Na2SO4, filtered, concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE: 30% - 40%) to give [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E5, 640.0 mg, eluted first) as a white solid. MS calculated value 921.3 (MH + ), measured value 921.4 (MH + ).

[0129] 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-propan-1-ol (Compound E6) [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) was added cesium fluoride (421.8 mg, 2.78 mmol). The mixture was stirred at 60 °C for 16 h. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE: 30% - 60%) to give 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E6, 360.0 mg) as a yellow oil. MS calculated 683.2 (MH + ), found 683.1 (MH + ).

[0130] 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-propan-1-ol (Compound E7) 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-propan-1-ol (Compound El6, 360.0 mg, 0.53 mmol), bis(pinacolato)diboron (200.6 mg, 0.79 mmol) in toluene (6 mL) was added potassium acetate (0.08 mg, 1.32 mmol) and Pd(dppf)Cl2 (40 mg, 0.1 mmol). The reaction mixture was degassed by nitrogen bubbling for 5 minutes and then stirred at 80 °C for 15 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE: 30% - 50%) to give 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E7, 300.0 mg) as a yellow gum. MS calculated value 731.4 (MH + ), measured value 731.4 (MH + ).

[0131] Step 7: Preparation of methyl (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]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylate (Compound E8) 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-propan-1-ol (Compound E7, 0.3 g, 0.41 mmol) and methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylate (Intermediate B, 196.7 mg, 0.41 mmol) in a mixture of toluene (3 mL) / 1,4-dioxane (1 mL) / water (1 mL) were added K3PO4 (221.3 mg, 1.04 mmol) and Pd(dtbpf)Cl2 (27.05 mg, 0.04 mmol). The mixture was stirred at 70 °C for 12 h under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE: 60% - 80%) to give methyl (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]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylate (Compound E8, 200.0 mg) as a yellow gum. MS calculated value 1001.4 (MH + ), measured value 1001.4 (MH + ).

[0132] 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]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound E9) To a mixture of methyl (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]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylate (Compound E8, 200.0 mg, 0.2 mmol) in DCE (5 mL) was added Me3SnOH (200.0 mg, 1.11 mmol). The mixture was stirred at 60 °C for 12 h. The reaction mixture was concentrated under vacuum to give a residue. EtOAc (10 mL) and water (10 mL) were added to the residue and the layers were separated. The aqueous phase was extracted with EtOAc (15 mL, 2 times). The combined organic phases 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]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound E9, 188.0 mg) as a brown solid. MS calculated 987.4 (MH + +), found 987.4 (MH + +).

[0133] Step 9: Preparation of tert-Butyl 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-Tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-Hexaen-7-yl]Carbamate (Compound E10) (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]Thiazol-2-yl]Propanoyl]Hexahydropyridazine-3-Carboxylic Acid (Compound E9, 188.0 mg, 0.19 mmol) in DCM (20 mL) was added with DIEA (0.7 mL, 3.81 mmol), EDCI (550.0 mg, 2.87 mmol) and HOBt (65.0 mg, 0.48 mmol) at 0 °C. After stirring at 20 °C for 12 h, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (20 mL, 3 times). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc in PE: 50% - 70%) to give tert-Butyl 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-Tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound E10, 110.0 mg) was obtained as a yellow solid. MS calculated value 969.4 (MH + ), measured value 969.5 (MH + ).

[0134] Step 10: (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate E) tert-Butyl 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26To a solution of [[ID=]], 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound E10, 110.0 mg, 0.11 mmol) in DCM (1 mL) was added TFA (1.0 mL, 12.98 mmol). The mixture was stirred at 20 °C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under vacuum to obtain a residue. Saturated aqueous NaHCO3 (20 mL) was added and the mixture was extracted with EtOAc (15 mL, 3 times). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and (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-tetrazapentacyclo[17.5.2.1 + ) was obtained as a yellow solid. MS calculated value 869.4 (MH + ), measured value 869.2 (MH

[0135] 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione Using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate, the title compound was prepared in the same manner as Intermediate E.

[0136] 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-tetrazapentacyclo-[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione TIFF2025522629000042.tif50170 This compound was prepared according to the following scheme. TIFF2025522629000043.tif159170TIFF2025522629000044.tif206170

[0137] Step 1: Preparation of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (Compound G1) To a mixture of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3, 30 g, 87.73 mmol) and morpholine (7.6 g, 87.73 mmol) in toluene (450 mL) were added Cs2CO3 (57.2 g, 175.45 mmol), (R)-binap (2.7 g, 4.39 mmol), and Pd(OAc)2 (0.98 g, 4.39 mmol). The reaction mixture was degassed and purged with nitrogen three times, and the mixture was stirred at 90 °C for 12 h under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography to give 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (Compound G1, 21 g) as a yellow oil. MS calculated value 301.1 (MH +) Measured value 301.1 (MH + )

[0138] Step 2: Preparation of 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]morpholine (Compound G2) To a solution of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (Compound G1, 21 g, 63.3 mmol), bis(pinacolato)diboron (24.0 g, 94.63 mmol) and KOAc (13.6 g, 138.79 mmol) in toluene (500 mL) was added Pd(dppf)Cl2 (4.4 g, 6.31 mmol). The mixture was degassed and purged with nitrogen three times, and the mixture was stirred at 90 °C for 12 hours under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo to give the crude product 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]morpholine (Compound G2, 45 g) as a yellow gum, which was used in the next step. MS calculated value 349.2 (MH + ) Measured value 349.2 (MH + )

[0139] Step 3: Preparation of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound G3) To a solution of 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]morpholine (Compound G2, 40.6 g, 46.65 mmol), [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 31 g, 46.65 mmol) in 1,4-dioxane (420 mL) / water (80 mL) was added K3PO4 (29.7 g, 2.33 mmol) and Pd(dppf)Cl2 (1.7 g, 0.29 mmol) was added. The mixture was degassed by nitrogen bubbling for 2 minutes, and the reaction mixture was stirred at 90 °C for 18 hours. After cooling to room temperature, the reaction mixture was extracted with EA (200 mL, 3 times). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography to give [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound G3, 17.2 g) as a yellow oil. MS calculated value 758.3 (MH + ) and measured value 758.3 (MH + ).

[0140] Step 4: Preparation of [3-[5-bromo-1-ethyl-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound G4) [3-[5-Bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound G3, 15 g, 19.77 mmol) in DMF (300 mL) was added with Cs2CO3 (19.3 g, 59.3 mmol) and iodomethane (6.16 g, 39.53 mmol) at 0 °C. After stirring at 20 °C for 16 h, the reaction mixture was poured into water (200 mL) and extracted with EtOAc (200 mL, 3 times). The combined organic layers were washed with brine (10 mL, 3 times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by column chromatography to give [3-[5-bromo-1-ethyl-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound G4, 14.7 g) as a yellow oil. MS calculated value 786.3 (MH + ), measured value 786.4 (MH + ).

[0141] Step 5: Preparation of 3-[5-bromo-1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G5) and 3-[5-bromo-1-ethyl-6-fluoro-(2P)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G6). [3-[5-Bromo-1-ethyl-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound G4, 14.7 g, 18.68 mmol) in DMF (160 mL) was added cesium fluoride (14.2 g, 93.41 mmol). The mixture was stirred at 60 °C for 48 h. After cooling to room temperature, EtOAc (300 mL) and water (300 mL) were added to the reaction mixture and the layers were separated. The aqueous phase was extracted with EtOAc (200 mL, 3 times). The combined organic layers were washed with brine (200 mL, 4 times), dried over Na2SO4, filtered and concentrated under vacuum to give a residue. This residue was purified by column chromatography to give 3-[5-bromo-1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G5, 6 g, eluted first) as a colorless foam and 3-[5-bromo-1-ethyl-6-fluoro-(2P)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G6, 4.5 g, eluted later) as a colorless foam. Compound G5: MS calculated 548.2 (MH + ), found 548.2 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.41 (d, J = 2.4 Hz, 1H), 7.92 (d, J = 6.8 Hz, 1H), 7.37 - 7.33 (m, 2H), 4.58 (s, 1H), 4.05 - 3.98 (m, 2H), 3.87-3.82 (m, 5H), 3.27 - 3.23 (m, 4H), 3.15 - 3.13 (m, 1H), 3.00 (s, 3H), 2.75-2.71 (m, 1H), 2.24 - 2.22 (m, 1H), 1.42 (d, J = 6.4 Hz, 3H), 1.22 (t, J = 7.2 Hz, 3H), 0.76 (s, 3H), 0.76 (s, 3H).

[0142] X-ray crystal analysis of Compound G5 The absolute configuration structure of Compound G5 was confirmed by X-ray crystal analysis of a single crystal. (Figure 1)

[0143] Step 6: Preparation of 3-[1-Ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G7) To a toluene (60 mL) solution of 3-[5-Bromo-1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G5, 6 g, 10.94 mmol) and bis(pinacolato)diboron (4.2 g, 16.41 mmol), potassium acetate (2.7 g, 27.35 mmol) and Pd(dppf)Cl2 (0.8 g, 1.09 mmol) were added. The reaction mixture was degassed by nitrogen bubbling for 5 minutes and then stirred at 90 °C for 15 hours. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column flash chromatography to give 3-[1-Ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G7, 4.5 g) as a colorless gum. MS calculated value 596.4 (MH + )、measured value 596.4 (MH + ).

[0144] Step 7: Preparation of Methyl (3S)-1-[(2S)-2-(tert-Butoxycarbonylamino)-3-[4-[1-Ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylate (Compound G8) To a mixture of 3-[1-Ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G7, 4.5 g, 7.56 mmol) and Methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylate (Intermediate B, 3.6 g, 7.56 mmol) in Toluene (45 mL) / 1,4-Dioxane (15 mL) / Water (15 mL) was added K3PO4 (4.0 g, 18.89 mmol) and Pd(dtbpf)Cl2 (492.5 mg, 0.75 mmol). The mixture was stirred at 70 °C for 12 h under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column flash chromatography to give Methyl (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylate (Compound G8, 3.8 g) as a colorless gum. MS Calcd 866.4 (MH + +), Found 866.4 (MH + +).

[0145] Step 8: Preparation of (3S)-1-[(2S)-2-(tert-Butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound G9) To a mixture of methyl (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylate (Compound G8, 3.8 g, 4.39 mmol) in DCE (76 mL) was added Me3SnOH (3.2 g, 17.55 mmol). The mixture was stirred at 60 °C for 48 h. The reaction mixture was concentrated under vacuum to give a residue. To this residue were added EtOAc (200 mL) and water (10 mL), and the layers were separated. The aqueous phase was extracted with EtOAc (200 mL, 2 times). The combined organic phases were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound G9, 3.7 g) as a brown solid. MS calculated 852.4 (MH + +), found 852.4 (MH + +).

[0146] Project 9: Preparation of tert-butyl 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,2 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound G10) (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound G9, 2.5 g, 2.93 mmol) in DCM (250 mL) was added with DIEA (7.58 mL, 58.68 mmol), EDCI (8.4 g, 44.01 mmol) and HOBt (991.2 mg, 0.91 mmol) at 0 °C. After stirring at 20 °C for 12 h, the reaction mixture was poured into water (100 mL) and extracted with EtOAc (100 mL, 3 times). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, concentrated under vacuum to obtain a residue, which was purified by column chromatography to give tert-butyl 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound G10, 1.2 g) was obtained as a yellow oil. MS calculated value 834.4 (MH + ), measured value 834.4 (MH + ).

[0147] Project 10: (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-tetrazapentacyclo-[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate G) tert-Butyl 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 To a solution of tert-butyl 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound G10, 1.2 g, 1.44 mmol) in DCM (12 mL) was added TFA (6.0 mL, 181.72 mmol). The mixture was stirred at 20 °C for 3 hours. After completion of the reaction, the reaction mixture was concentrated under vacuum to give a residue. Saturated aqueous NaHCO3 (60 mL) was added and the mixture was extracted with EtOAc (80 mL, 3 times). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and (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-tetrazapentacyclo[17.5.2.1 +) Measurement value 734.3 (MH + )。

[0148] 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione TIFF2025522629000045.tif561705-Bromo-6-fluoro-1H-indole (Compound C2) was replaced with 5-bromo-4-fluoro-1H-indole, and the title compound was prepared in the same manner as the preparation of Intermediate C.

[0149] Intermediate I cis-tert-Butyl 3-ethynylcyclobutanecarboxylate TIFF2025522629000046.tif34170This compound was prepared according to the following scheme. TIFF2025522629000047.tif77170

[0150] Step 1: Preparation of cis-O1-tert-butyl O3-methylcyclobutane-1,3-dicarboxylate (Compound I2) A solution of Boc2O (37.9 g, 173.89 mmol) in tert-butanol (50 mL) was added dropwise to a mixture of cis-3-methoxycarbonylcyclobutanecarboxylic acid (Compound I1, 25.0 g, 158.08 mmol) and DMAP (38.6 g, 316.16 mmol) in tert-butanol (450 mL). After stirring at 25 °C for 0.5 h, the reaction mixture was diluted with water (150 mL) and extracted with EtOAc (150 mL, 3 times). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The resulting residue was purified by silica column (EtOAc in PE = 5% - 10%) to obtain cis-O1-tert-butyl O3-methylcyclobutane-1,3-dicarboxylate (Compound I2, 29.0 g) as a colorless liquid. MS calculated value 215.1 (MH + ), measured value 215.1 (MH + ).

[0151] Step 2: Preparation of cis-tert-butyl 3-(hydroxymethyl)-cyclobutanecarboxylate (Compound I3) Lithium borohydride (9.2 g, 420.05 mmol) was added to a mixture of cis-O1-tert-butyl O3-methylcyclobutane-1,3-dicarboxylate (Compound I2, 29.0 g, 140.02 mmol) in THF (290 mL) at 0 °C under a nitrogen atmosphere. After stirring at 25 °C for 2 h, EtOAc (800 mL) and water (200 mL) were added to the reaction mixture and the layers were separated. The aqueous phase was extracted with EtOAc (300 mL, 3 times). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by silica column (EtOAc in PE = 10% - 30%) to obtain cis-tert-butyl 3-(hydroxymethyl)-cyclobutanecarboxylate (Compound I3, 19 g) as a colorless liquid. MS calculated value 186.1 (MH + ), measured value 186.1 (MH + ).

[0152] Step 3: Preparation of cis-tert-butyl 3-formylcyclobutanecarboxylate (Compound I4) To a mixture of cis-tert-butyl 3-(hydroxymethyl)-cyclobutanecarboxylate (Compound I3, 19.0 g, 102.01 mmol) in DCM (230 mL) was added DMAP (51.9 g, 122.42 mmol) at 0 °C. After stirring at 25 °C for 1 h, the reaction mixture was filtered. The filtrate was concentrated in vacuo and purified by silica column (EtOAc in PE = 5% - 20%) to give cis-tert-butyl 3-formylcyclobutanecarboxylate (Compound I4, 15.0 g) as a colorless oil. MS calculated value 184.1 (MH + ), measured value 184.1 (MH + ).

[0153] Step 4: Preparation of cis-tert-butyl 3-ethynylcyclobutanecarboxylate (Intermediate I) To a mixture of cis-tert-butyl 3-formylcyclobutanecarboxylate (Compound I4, 15.0 g, 81.42 mmol) and potassium carbonate (22.5 g, 162.84 mmol) in methanol (200 mL) was added dimethyl (1-diazo-2-oxopropyl)-phosphonate (Compound I5, 23.5 g, 122.13 mmol) at 0 °C. The mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated in vacuo to remove the solvent, diluted with water (100 mL), and extracted with ethyl acetate (100 mL, 2 times). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The obtained residue was purified by silica gel chromatography to give cis-tert-butyl 3-ethynylcyclobutanecarboxylate (Intermediate I, 7 g) as a colorless oil. MS calculated value 180.1 (MH + ), measured value 180.1 (MH + ).

[0154] Intermediate J (7S,13S)-7-Amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione TIFF2025522629000048.tif491704 - The title compound was prepared in the same manner as for the preparation of Intermediate G, using 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (Compound A1) instead of 491704-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (Compound G1).

[0155] Intermediate K (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione TIFF2025522629000049.tif49170 The title compound was prepared in the same manner as for the preparation of Intermediate G, using 2,2,2-trifluoroethyl trifluoromethanesulfonate instead of iodoethane.

[0156] Example 1 trans-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.12,5 .1 9,13 .0 22,26 [[Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide Compound TIFF2025522629000050 was prepared according to the following scheme: TIFF2025522629000051.tif62170

[0157] Step 1: trans-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Preparation of [[Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I, 70.0 mg, 0.23 mmol), (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-tetrazapentacyclo[17.5.2. 12,5 .1 9 . 13 .0 22,26To a solution of octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (intermediate C, 100.0 mg, 0.12 mmol), EDCI (50.0 mg, 0.26 mmol) and DIEA (0.1 mL, 0.62 mmol) in DMF (2 mL) was added HOBT (34.0 mg, 0.25 mmol) at 0 °C. After stirring at 16 °C for 1 hour, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (20 mL, 3 times). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under vacuum to give a residue. The obtained residue was purified by preparative HPLC to give N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide (Example 1, 34.8 mg) was obtained as an off-white solid. MS calculated value 1088.5 (MH + ), measured value 1088.5 (MH + ). 11H NMR (400 MHz, METHANOL-d4) δ = 8.68 (d, J = 7.2 Hz, 1H), 8.50 (d, J = 3.2 Hz, 1H), 7.69 (d, J = 2.4 Hz, 1H), 7.54 - 7.43 (m, 2H), 5.72 - 5.65 (m, 1H), 5.26 - 5.10 (m, 1H), 4.82 - 4.75 (m, 1H), 4.46 - 4.38 (m, 1H), 4.27 - 4.18 (m, 2H), 4.12 - 3.86 (m, 2H), 3.82 - 3.56 (m, 5H), 3.54 - 3.41 (m, 2H), 3.37 - 3.33 (m, 4H), 3.30 - 3.23 (m, 2H), 3.21 - 3.06 (m, 2H), 3.02 - 2.91 (m, 6H), 2.87 - 2.68 (m, 3H), 2.65 - 2.16 (m, 6H), 2.05 - 1.55 (m, 4H), 1.45 (d, J = 6.0 Hz, 3H), 1.35 - 1.25 (m, 1H), 1.03 - 0.94 (m, 5H), 0.91 - 0.82 (m, 3H), 0.44 (s, 3H) ppm.

[0158] Compound 1I was prepared according to the following scheme. TIFF2025522629000052.tif145170

[0159] Step 1: Preparation of methyl 3-(methoxymethylene)cyclobutanecarboxylate (Compound 1B) A solution of (methoxymethyl)triphenylphosphonium chloride (267.5 g, 780.46 mmol) in THF (1.6 L) was slowly added with potassium tert-butoxide (87.6 g, 780.46 mmol) at 0 °C and warmed to 20 °C. After 1.5 h, methyl 3-oxocyclobutanecarboxylate (Compound 1A, 50.0 g, 390.23 mmol) was added to the reaction mixture. After stirring at 70 °C for 3 h, the reaction mixture was concentrated under vacuum to obtain a residue. A mixed solution of PE in EtOAc (10:1, 1.1 L) was added to the residue. After stirring at 20 °C for 0.5 h, the suspension was filtered and the filtrate was concentrated under vacuum to obtain a residue. The residue was purified by column chromatography (EtOAc in PE = 0% - 10%) to obtain methyl 3-(methoxymethylene)cyclobutanecarboxylate (Compound 1B, 18.0 g) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d) δ : 5.85 - 5.79 (m, 1 H), 3.70 (s, 3 H), 3.60 (s, 3 H), 3.29 - 3.09 (m, 1 H), 3.01 - 2.90 (m, 2H), 2.89 - 2.71 (m, 2 H) ppm.

[0160] Step 2: Preparation of methyl 3-formylcyclobutanecarboxylate (Compound 1C) TFA (26.0 mL) was added to a solution of methyl 3-(methoxymethylene)cyclobutanecarboxylate (Compound 1B, 26.0 g, 166.47 mmol) in DCM (300 mL) / water (30 mL). The reaction mixture was stirred at 20 °C for 3 h. After completion of the reaction, H2O (600 mL) was added to the reaction mixture, followed by extraction with DCM (100 mL, 3 times). The organic layer was washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain methyl 3-formylcyclobutanecarboxylate (Compound 1C, 18.0 g, 126.63 mmol) as a yellow oil. 11H NMR (400 MHz, CHLOROFORM-d) δ: 9.84 - 9.52 (m, 1 H), 3.75 - 3.63 (m, 3 H), 3.32 - 3.20 (m, 1 H), 3.18 - 3.07 (m, 1 H), 2.67 - 2.38 (m, 4 H) ppm.

[0161] Step 3: Preparation of methyl 3-ethynylcyclobutanecarboxylate (Compound 1D) A solution of methyl 3-formylcyclobutanecarboxylate (Compound 1C, 10.0 g, 70.35 mmol) in methanol (120 mL) was cooled to 0 °C, and dimethyl (1-diazo-2-oxopropyl)phosphonate (21.0 g, 109.31 mmol) and potassium carbonate (20.0 g, 144.71 mmol) were added to the reaction mixture. After stirring at 20 °C for 3 h, H2O (150 mL) was added to the reaction mixture, and then it was extracted with PE (60 mL, 2 times). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by column chromatography (EtOAc in PE = 0% - 25%) to obtain methyl 3-ethynylcyclobutanecarboxylate (Compound 1D, 6.0 g) as a colorless oil. 1 1H NMR (400 MHz, CHLOROFORM-d) δ: 3.75 - 3.62 (m, 3 H), 3.42 - 3.21 (m, 1 H), 3.07 - 2.89 (m, 1 H), 2.65 - 2.33 (m, 4 H), 2.23 - 2.17 (m, 1 H) ppm.

[0162] Step 4: Preparation of 3-ethynylcyclobutanecarboxylic acid (Compound 1E) To a solution of methyl 3-ethynylcyclobutanecarboxylate (Compound 1D, 6.0 g, 43.43 mmol) in THF (10 mL) / water (30 mL) was added lithium hydroxide (3.6 g, 86.86 mmol) at 0 °C, and then the solution was stirred at 20 °C for 3 h. After completion of the reaction, the reaction mixture was concentrated under vacuum to remove THF, to which H2O (60 mL) was added, and the mixture was extracted with MTBE (30 mL). The MTBE phase was discarded, the pH of the aqueous phase was acidified to pH = 5 with HCl aqueous solution (1 N, 60 mL), and it was extracted with EtOAc (60 mL, 3 times). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain 3-ethynylcyclobutanecarboxylic acid (Compound 1E, 3.8 g) as a colorless oil. 1 H NMR (400 MHz, CHLOROFORM-d) δ: 12.14 - 9.87 (m, 1 H), 3.36 - 3.15 (m, 1 H), 3.10 - 2.95 (m, 1 H), 2.68 - 2.53 (m, 2 H), 2.51 - 2.35 (m, 2 H), 2.22 (dd, J = 15.2, 2.4 Hz, 1 H) ppm.

[0163] Step 5: Preparation of trans-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1F) and cis-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1G) To a solution of 3-ethynylcyclobutanecarboxylic acid (Compound 1E, 3.8 g, 30.61 mmol) in DMF (50 mL) were added DIEA (19.0 mL, 114.96 mmol) and HATU (14.3 g, 37.48 mmol). After stirring at 0 °C for 10 minutes, tert-butyl (2S)-3-methyl-2-(methylamino)butanoate (5.7 g, 30.44 mmol) was added to the reaction mixture. The reaction mixture was stirred at 0 °C for an additional 1 hour. After completion of the reaction, H2O (120 mL) was added to the reaction mixture, and the mixture was extracted with EtOAc (40 mL, 3 times). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by column chromatography (EtOAc in PE: 9% - 16%) and preparative HPLC (column: Welch Ultimate XB-CN 250×50×10 μm; mobile phase: hexane - EtOH (0.1% FA); B%: 1% - 20%, 15 minutes) to give cis-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1G, eluted first, 3 g) as a yellow oil and trans-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1F, eluted later, 2 g) as a yellow oil.

[0164] cis-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1G, peak 1). MS calculated value 294.2 (MH + ), measured value 294.1 (MH + ). 11H NMR (400 MHz, CHLOROFORM-d) δ = 4.80 (d, J = 10.4 Hz, 0.5 H), 3.58 (d, J = 10.8 Hz, 0.5 H), 3.27 - 3.11 (m, 1 H), 3.01 - 2.91 (m, 1 H), 2.87 (d, J = 6.8 Hz, 3 H), 2.59 - 2.40 (m, 4 H), 2.25 - 2.12 (m, 2 H), 1.45 (s, 9 H), 1.00 (dd, J = 14.4, 6.4 Hz, 3 H), 0.84 (dd, J = 6.8, 1.2 Hz, 3 H) ppm. The stereochemistry of Compound 1G was confirmed by 2D-NMR.

[0165] trans-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1F, Peak 2). MS calculated value 294.2 (MH + ), measured value 294.1 (MH + ). 1 1H NMR (400 MHz, CHLOROFORM-d) δ = 4.80 (d, J = 10.4 Hz, 0.5 H), 3.69 - 3.46 (m, 1.5 H), 3.11 - 3.01 (m, 1 H), 2.88 (d, J = 3.2 Hz, 3 H), 2.73 - 2.60 (m, 2 H), 2.40 - 2.27 (m, 2 H), 2.25 - 2.11 (m, 2 H), 1.45 (d, J = 2.8 Hz, 9 H), 1.01 (dd, J = 12.0, 6.8 Hz, 3 H), 0.84 (dd, J = 6.8, 1.6 Hz, 3 H) ppm. The stereochemistry of Compound 1F was confirmed by 2D-NMR.

[0166] Step 6: Preparation of trans-tert-butyl (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoate (Compound 1H) A suspension of CuI (408.9 mg, 2.15 mmol), K2CO3 (593.5 mg, 4.29 mmol) and TMEDA (249.5 mg, 2.15 mmol) in DMF (10 mL) was stirred at 25 °C for 20 min under an argon atmosphere. TMSCF3 (407.1 mg, 2.86 mmol) was added to the reaction, and the reaction mixture was stirred for 10 min under an argon atmosphere. A solution of TMSCF3 (407.09 mg, 2.86 mmol) and trans-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1F, 420.0 mg, 1.43 mmol) in DMF (10 mL) was added to the reaction. The reaction mixture was stirred at 0 °C for 30 min and warmed to 25 °C. After stirring at 25 °C for an additional 12 h, H2O (30 mL) was added to the reaction mixture, which was then extracted with EtOAc (10 mL, 3 times). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by reverse-phase chromatography and preparative HPLC to give trans-tert-butyl (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoate (Compound 1H, 80.0 mg) as a yellow oil. MS calculated 362.2 (MH + ), found 362.1 (MH + ). 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.80 (d, J =10.0 Hz, 0.5 H), 3.62 - 3.46 (m, 1.5 H), 3.28 - 3.13 (m, 1 H), 2.89 (d, J = 4.4 Hz, 3 H), 2.82 - 2.67 (m, 2 H), 2.48 - 2.38 (m, 2 H), 2.29 - 2.15 (m, 1 H), 1.46 (d, J = 2.8 Hz, 9 H), 1.05 - 0.98 (m, 3 H), 0.85 (d, J = 6.8 Hz, 3 H) ppm.

[0167] Step 7: Preparation of trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) To a solution of trans-tert-butyl (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoate (Compound 1H, 80.0 mg, 0.22 mmol) in DCM (1 mL) was added TFA (1.0 mL), and the mixture was stirred at 20 °C for 1 h. After completion of the reaction, the reaction mixture was concentrated under vacuum to give trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I, 80.0 mg) as a yellow oil, which was used directly in the next step. MS calculated value 306.0 (MH + ), measured value 306.0 (MH + ).

[0168] Example 2 N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide Instead of TIFF2025522629000053.tif63170trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I), (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E) was used to prepare the title compound in the same manner as in the preparation of Example 1. Example 2 (78.9 mg) was obtained as a yellow solid. MS calculated value 1089.5 (MH + ), measured value 1089.7 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.68 (d, J = 7.2 Hz, 1 H), 8.50 (d, J = 2.8 Hz, 1 H), 7.70 (d, J = 2.0 Hz, 1 H), 7.51 (d, J = 2.4 Hz, 1 H), 7.48 (d, J = 12.8 Hz, 1 H), 5.71 (t, J = 8.8 Hz, 1 H), 5.23 - 5.13 (m, 1 H), 4.47 - 4.38 (m, 2 H), 4.37 - 4.11 (m, 6 H), 4.10 - 3.93 (m, 3 H), 3.81 - 3.77 (m, 1 H), 3.76 - 3.59 (m, 4 H), 3.47 (d, J = 14.8 Hz, 1 H), 3.35 (s, 3 H), 3.29 - 3.23 (m, 2 H), 3.19 - 3.10 (m, 2 H), 2.99 (s, 3 H), 2.87 (s, 3 H), 2.84 - 2.79 (m, 1 H), 2.57 (d, J = 14.4 Hz, 1 H), 2.25 - 2.16 (m, 2 H), 2.01 - 1.92 (m, 1 H), 1.87 - 1.77 (m, 1 H), 1.70 - 1.58 (m, 1 H), 1.45 (d, J = 6.0 Hz, 3 H), 1.39 - 1.25 (m, 1 H), 1.00 - 0.85 (m, 10 H), 0.44 (s, 3 H) ppm.

[0169] Compound 2E was prepared according to the following scheme. TIFF2025522629000054.tif119170

[0170] Step 1: Preparation of 3-ethynylazetidine (Compound 2B) To a solution of tert-butyl 3-ethynylazetidine-1-carboxylate (Compound 2A, 3.5 g, 19.31 mmol) in DCM (36 mL) was added TFA (17.7 g, 155.76 mmol). The reaction mixture was stirred at 20 °C for 1 h. After completion of the reaction, the reaction mixture was evaporated and co-evaporated three times with DCM (20 mL) to give 3-ethynylazetidine (Compound 2B, 3.5 g, crude, TFA salt) as a yellow oil, which was used directly in the next step without purification.

[0171] Step 2: Preparation of tert-butyl (2S)-2-[(3-ethynylazetidine-1-carbonyl)-methyl-amino]-3-methyl-butanoate (Compound 2C) To a mixture of tert-butyl (2S)-3-methyl-2-(methylamino)butanoate (3.7 g, 19.76 mmol) in DCM (50 mL) were added DIEA (8.5 mL, 48.8 mmol) and triphosgene (2.1 g, 7.08 mmol). After stirring at 0 °C for 10 min, a mixture of 3-ethynylazetidine;2,2,2-trifluoroacetic acid (Compound 2B, 3.5 g, 17.94 mmol) and DIEA (13.0 mL, 74.64 mmol) in DCM (50 mL) was added to the reaction. The resulting mixture was stirred at 20 °C for an additional 1 h. After completion of the reaction, saturated aqueous NaHCO3 solution (500 mL) was added to the reaction mixture, and the mixture was extracted with EtOAc (100 mL, 2 times). The combined organic layers were washed with brine (600 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by column chromatography (EtOAc in PE = 11% - 25%) to give tert-butyl (2S)-2-[(3-ethynylazetidine-1-carbonyl)-methyl-amino]-3-methyl-butanoate (Compound 2C, 3.2 g) as a yellow oil. MS calculated value 239.2 (M - C4H9 + H+ ) Measured value 239.0 (M-C4H9+H + )。

[0172] Step 3: Preparation of tert-butyl (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoate (Compound 2D) A mixture of CuI (1.5 g, 8.15 mmol), potassium carbonate (2.3 g, 16.34 mmol) and TMEDA (947.3 mg, 8.15 mmol) in DMF (10 mL) was stirred at 20 °C for 20 minutes under an argon atmosphere. TMSCF3 (1.5 g, 10.87 mmol) was added to the reaction mixture and stirred at 20 °C for 20 minutes. A mixture of tert-butyl (2S)-2-[(3-ethynylazetidine-1-carbonyl)-methyl-amino]-3-methyl-butanoate (Compound 2C, 1.6 g, 5.43 mmol) and TMSCF3 (1.5 g, 10.87 mmol) in DMF (30 mL) was added to the reaction mixture. After stirring at 20 °C for an additional 12 hours under an argon atmosphere, H2O (100 mL) was added to the reaction mixture, and then extracted with EtOAc (30 mL, 3 times). The combined organic layers were washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by column chromatography (EtOAc in PE: 11% - 25%) and concentrated under vacuum to obtain a residue. The residue was re-purified by reverse-phase HPLC to obtain tert-butyl (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoate (Compound 2D, 600.0 mg) as a yellow oil. MS calculated value 307.2 (M-C4H9+H + ) Measured value 307.1 (M-C4H9+H + )。

[0173] Step 4: Preparation of (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E) To a solution of tert-butyl (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoate (Compound 2D, 300.0 mg, 0.83 mmol) in DCM (2 mL) was added TFA (2.6 g, 23.36 mmol). The reaction mixture was stirred at 20 °C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under vacuum to obtain a residue. The residue was co-evaporated with DCM (6 mL, 3 times) to give (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E, 250.0 mg) as a yellow oil, which was used directly in the next step without purification. MS calculated value 307.1 (MH + ), measured value 307.0 (MH + ).

[0174] Example 3 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide Instead of TIFF2025522629000055.tif68170trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I), cis-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 3B) was used to prepare the title compound in the same manner as in the preparation of Example 1. Example 3 (202.7 mg) was obtained as a yellow solid. MS calculated value 1088.5 (MH + ), measured value 1088.5 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.68 (d, J = 7.6 Hz, 1H), 8.50 (d, J = 2.8 Hz 1H), 8.67 (d, J = 2.4 Hz 1H), 7.52 - 7.43 (m, 2H), 5.74 - 5.61 (m, 1H), 5.21 - 5.12 (m, 1H), 4.94 - 4.86 (m, 2H), 4.83 - 4.81 (m, 1H), 4.78 (d, J = 11.2 Hz, 1H), 4.46 - 4.39 (m, 1H), 4.26 - 4.19 (m, 2H), 4.11 - 3.90 (m, 2H), 3.84 - 3.64 (m, 4H), 3.62 - 3.43 (m, 4H), 3.38 - 3.33 (m, 4H), 3.18 - 3.08 (m, 2H), 3.00 (s, 3H), 2.98 - 2.93 (m, 3H), 2.84 - 2.75 (m, 1H), 2.74 - 2.63 (m, 2H), 2.61 - 2.44 (m, 3H), 2.27 - 2.16 (m, 2H), 2.01 - 1.93 (m, 1H), 1.86 - 1.76 (m, 1H), 1.70 - 1.59 (m, 1H), 1.45 (d, J = 6.0 Hz, 3H), 1.10-1.00 (m, 1H), 1.00 - 0.95 (m, 5H), 0.90 - 0.83 (m, 3H), 0.44 (s, 3H) ppm.

[0175] Instead of trans-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1F), cis-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1G) was used to prepare Compound 3B in the same manner as the preparation of Compound 1I.

[0176] Example 4 N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)benzamide The TIFF2025522629000056.tif52170 compound was prepared according to the following scheme: TIFF2025522629000057.tif203170

[0177] Step 1: tert-Butyl N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Preparation of [[(7S,13S)-7-amino-24-fluoro-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[17.5.2.111,15.02,6.114,18]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate (Compound 4A) To a mixture of BOC-N-ME-VAL-OH (93.8 mg, 0.41 mmol) and DIEA (0.2 mL, 0.94 mmol) in DMF (1 mL) was added HATU (154.3 mg, 0.41 mmol) and [[(7S,13S)-7-amino-24-fluoro-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[17.5.2.111,15.02,6.114,18]octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C, 250.0 mg, 0.31 mmol). After stirring at 15 °C for 1 h, the reaction mixture was purified by reverse phase chromatography to give tert-butyl N-[(1S)-1-[[(7S,13S)-24-fluoro-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[17.5.2.111,15.02,6.114,18]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate (Compound 4A, 300.0 mg) as a yellow solid. MS calculated 1014.4 (MH 2,5 .1 9,13 .0 22,26 ), and [[(7S,13S)-7-amino-24-fluoro-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[17.5.2.111,15.02,6.114,18]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate (Compound 4A, 300.0 mg) was obtained as a yellow solid. MS calculated 1014.4 (MH 2,5 .1 9,13 .0 22,26 ), measured 1014.3 (MH + ). + )

[0178] Process 2: (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(methylamino)butanamide (Compound 4B) Preparation tert-Butyl N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate (Compound 4A, 300.0 mg, 0.3 mmol) in DCM (6 mL) mixture, TFA (4.0 mL) was added. After stirring at 15 °C for 0.5 h, the reaction mixture was concentrated in vacuo to give a residue. The obtained residue was diluted with saturated aqueous NaHCO3 (30 mL) and extracted with EtOAc (20 mL, 3 times). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, concentrated in vacuo, and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26[[Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(methylamino)butanamide (Compound 4B, 270.0 mg) was obtained as a yellow oil and used as such in the next step. MS calculated value 914.4 (MH + ), measured value 914.3 (MH + ).

[0179] Step 3: Preparation of N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)benzamide (Example 4) (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26A solution of [[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)benzamide (Example 4, 9.8 mg) was obtained as an off-white solid. MS calculated value 1110.4 (MH + +), measured value 1110.6 (MH + +). 11H NMR (400 MHz, Methanol-d4) δ = 8.69 (d, J = 9.2 Hz, 1H), 8.47 - 8.43 (m, 1H), 7.79 (d, J = 8.4 Hz, 2H), 7.73 - 7.69 (m, 1H), 7.58 (d, J = 8.0 Hz, 2H), 7.49 - 7.44 (m, 1H), 7.39 - 7.33 (m, 1H), 5.78 - 5.71 (m, 1H), 5.19 - 5.10 (m, 1H), 4.60 - 4.41 (m, 1H), 4.32 - 4.15 (m, 2H), 3.80 - 3.70 (m, 1H), 3.53 - 3.40 (m, 5H), 3.23 - 3.08 (m, 4H), 2.97 (s, 3H), 2.95 - 2.90 (m, 3H), 2.84 - 2.71 (m, 2H), 2.62 - 2.52 (m, 4H), 2.46 - 2.14 (m, 3H), 2.08 - 1.53 (m, 4H), 1.43 (d, J = 6.0 Hz, 3H), 1.29 (s, 3H), 1.06 (t, J = 6.4 Hz, 5H), 0.97 (s, 3H), 0.48 - 0.41 (m, 3H) ppm.

[0180] Compound 4B was prepared according to the following scheme. TIFF2025522629000058.tif29170

[0181] Step 1: Preparation of methyl 4-(3,3,3-trifluoroprop-1-ynyl)benzoate (Compound 8B) A suspension of copper(I) iodide (1.8 g, 9.37 mmol), potassium carbonate (2.6 g, 18.77 mmol), and TMEDA (1.1 g, 9.37 mmol) in DMF (10 mL) was stirred at 20 °C under an argon atmosphere. After stirring for 20 minutes, TMSCF3 (1.8 g, 12.49 mmol) was added to the reaction mixture. A mixture of methyl 4-ethynylbenzoate (Compound 4D, 1.0 g, 6.24 mmol) and TMSCF3 (1.8 g, 12.49 mmol) in DMF (10 mL) was slowly added to the reaction mixture, and then the mixture was stirred at 20 °C for an additional 12 hours under an argon atmosphere. After completion of the reaction, H2O (200 mL) was added to the reaction mixture, and the mixture was extracted with EtOAc (80 mL, 3 times). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The obtained residue was purified by column chromatography (SiO2, EtOAc in PE = 0% - 95%) to obtain methyl 4-(3,3,3-trifluoroprop-1-ynyl)benzoate (Compound 4D, 233.0 mg) as a colorless solid. 1 H NMR (400 MHz, CHLOROFORM-d) δ = 8.08 (d, J = 8.4Hz, 2H), 7.64 (d, J = 8.4Hz, 2H), 3.95 (s, 3H) ppm.

[0182] Step 2: Preparation of 4-(3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C) Lithium hydroxide monohydrate (69.5 mg, 1.66 mmol) was added to a solution of methyl 4-(3,3,3-trifluoroprop-1-ynyl)benzoate (Compound 4D, 180.0 mg, 0.79 mmol) in THF (1 mL) / water (1 mL). The mixture was stirred at 20 °C for 1 hour. After completion of the reaction, the pH of the reaction mixture was adjusted to 6 with 1M aqueous HCl, and the mixture was extracted with EtOAc (10 mL, 3 times). The combined organic layers were concentrated under vacuum to obtain 4-(3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C, 168.0 mg) as a white solid, which was used as is in the next step.

[0183] Example 5 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(2-pyrimidin-2-ylethynyl)cyclobutanecarboxamide TIFF2025522629000059.tif63170Instead of trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I), (2S)-3-methyl-2-[methyl-[cis-3-(2-pyrimidin-2-ylethynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 5B) was used to prepare the title compound in the same manner as in the preparation of Example 1. Example 4 (1.3 mg) was obtained as a yellow solid. MS calculated value 1098.5 (MH + ), measured value 1098.5 (MH + ). 11H NMR (400 MHz, CHLOROFORM-d) δ = 8.74 - 8.64 (m, 2H), 8.52 (d, J = 2.8 Hz, 1H), 7.72 - 7.61 (m, 1H), 7.55 - 7.46 (m, 1H), 7.25 - 7.20 (m, 1H), 7.18 - 7.09 (m, 2H), 5.74 - 5.62 (m, 1H), 5.13 - 4.88 (m, 1H), 4.79 - 4.68 (m, 1H), 4.63 - 4.41 (m, 2H), 4.35 - 4.16 (m, 2H), 4.13 - 3.93 (m, 1H), 3.89 - 3.77 (m, 2H), 3.77 - 3.69 (m, 2H), 3.66 - 3.53 (m, 3H), 3.52 - 3.46 (m, 1H), 3.39 - 3.33 (m, 3H), 3.32 - 3.21 (m, 2H), 3.20 - 3.07 (m, 3H), 2.92 - 2.87 (m, 3H), 2.87 - 2.82 (m, 2H), 2.80 (s, 1H), 2.73 - 2.61 (m, 3H), 2.43 - 2.37 (m, 1H), 2.36 - 2.29 (m, 1H), 2.26 - 2.20 (m, 1H), 2.14 (br s, 1H), 2.05 - 2.00 (m, 1H), 1.98 - 1.89 (m, 4H), 1.60 - 1.57 (m, 1H), 1.49 - 1.45 (m, 3H), 1.38 - 1.35 (m, 1H), 1.28 - 1.25 (m, 3H), 1.17 (br dd, J = 6.9, 19.4 Hz, 2H), 1.06 (br d, J = 6.5 Hz, 2H), 1.11 - 0.76 (m, 5H) ppm.

[0184] Compound 5B was prepared according to the following scheme. TIFF2025522629000060.tif41170

[0185] Step 1: Preparation of tert-butyl (2S)-3-methyl-2-[methyl-[cis 3-(2-pyrimidin-2-ylethynyl)cyclobutanecarbonyl]amino]butanoate (Compound 5A) To a solution of cis-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1G, 100.0 mg, 0.34 mmol) in THF (1 mL) were added triethylamine (0.2 mL, 1.02 mmol), 2-iodopyrimidine (70.2 mg, 0.34 mmol), tetrakis(triphenylphosphine)palladium(0) (39.4 mg, 0.03 mmol), and CuI (6.5 mg, 0.03 mmol). The reaction mixture was degassed and purged with nitrogen three times, and then stirred at 50 °C for 1 hour. After completion of the reaction, H2O (60 mL) was added to the reaction mixture, and the mixture was extracted with EtOAc (40 mL, twice). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by preparative HPLC to give tert-butyl (2S)-3-methyl-2-[methyl-[cis 3-(2-pyrimidin-2-ylethynyl)cyclobutanecarbonyl]amino]butanoate (Compound 5A, 60 mg) as a yellow oil. MS calculated value 372 (MH + ), measured value 372 (MH + ).

[0186] Step 2: Preparation of (2S)-3-methyl-2-[methyl-[cis-3-(2-pyrimidin-2-ylethynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 5B) A solution of tert-butyl (2S)-3-methyl-2-[methyl-[cis 3-(2-pyrimidin-2-ylethynyl)cyclobutanecarbonyl]amino]butanoate (Compound 5A, 60.0 mg, 0.16 mmol) in DCM (1 mL) was treated with TFA (0.2 mL). After stirring at 20 °C for 1 h, the reaction mixture was concentrated in vacuo to afford (2S)-3-methyl-2-[methyl-[cis-3-(2-pyrimidin-2-ylethynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 5B, 50 mg). The crude product was used in the next step. MS calculated 316 (MH + ), found 316 (MH + ).

[0187] Example 6 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide TIFF2025522629000061.tif60170trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Instead of octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C), cis-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 3B) 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) was used to prepare the title compound in the same manner as in the preparation of Example 1. Example 6 (32.2 mg) was obtained as a yellow solid. MS calculated value 1034.5 (MH + ), measured value 1034.5 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 8.66 (d, J = 7.6 Hz, 1 H), 8.48 (d, J = 2.8 Hz, 1 H), 7.62 (d, J = 2.4 Hz, 1 H), 7.52 (d, J = 2.8 Hz, 1 H), 7.33 (d, J = 12.8 Hz, 1 H), 5.78 - 5.67 (m, 1 H), 4.78 - 4.74 (m, 2 H), 4.48 - 4.36 (m, 1 H), 4.33 - 4.26 (m, 1 H), 4.24 - 3.96 (m, 4 H), 3.81 - 3.64 (m, 3 H), 3.56 - 3.42 (m, 3 H), 3.34 (s, 4 H), 3.29 - 3.24 (m, 3 H), 3.12 - 2.89 (m, 8 H), 2.89 - 2.80 (m, 1 H), 2.80 - 2.31 (m, 6 H), 2.31 - 2.12 (m, 2 H), 1.98 - 1.87 (m, 1 H), 1.87 - 1.75 (m, 1 H), 1.75 - 1.54 (m, 1 H), 1.43 (d, J = 6.0 Hz, 3 H), 1.08 - 0.88 (m, 9 H), 0.85 (d, J = 6.4 Hz, 3 H), 0.58 - 0.41 (m, 3 H) ppm.

[0188] Example 7 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide TIFF2025522629000062.tif66170trans-(2S)-3-Methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Instead of octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C), cis-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 3B) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Using octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate E), the title compound was prepared in the same manner as in the preparation of Example 1. Example 7 (30.8 mg) was obtained as a white solid. MS calculated value 1156.5 (MH + ), measured value 1157.1 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 8.71 (d, J = 7.6 Hz, 1 H), 8.41 (d, J = 2.8 Hz, 1 H), 7.67 (d, J = 2.4 Hz, 1 H), 7.58 (s, 1 H), 7.47 (d, J = 12.4 Hz, 1 H), 5.70 (t, J = 8.8 Hz, 1 H), 5.24 - 5.06 (m, 2 H), 4.80 - 4.74 (m, 2 H), 4.48 - 4.35 (m, 1 H), 4.30 - 4.15 (m, 2 H), 3.80 - 3.69 (m, 2 H), 3.62 - 3.47 (m, 2 H), 3.47 - 3.39 (m, 5 H), 3.38 - 3.35 (m, 3 H), 3.22 - 3.11 (m, 3 H), 2.98 - 2.82 (m, 8 H), 2.72 - 2.60 (m, 3 H), 2.51 - 2.46 (m, 1 H), 2.28 - 2.16 (m, 2 H), 2.01 - 1.91(m, 1 H), 1.89 - 1.74 (m, 1 H), 1.69 - 1.58 (m, 1 H), 1.46 (d, J = 6.0 Hz, 3 H), 1.10 - 1.01 (m, 1 H), 1.01 - 0.83 (m, 9 H), 0.49 (s, 3 H) ppm.

[0189] Example 8 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide TIFF2025522629000063.tif63170trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Instead of octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C), cis-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 3B) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Using octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate F), the title compound was prepared in the same manner as in the preparation of Example 1. Example 8 (23.3 mg) was obtained as a white solid. MS calculated value 1102.5 (MH + ), measured value 1102.7 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 8.65 (d, J = 7.2 Hz, 1 H), 8.40 (d, J = 2.8 Hz, 1 H), 7.61 (d, J = 2.4 Hz, 1 H), 7.33 - 7.27 (m, 2 H), 5.90 - 5.68 (m, 1 H), 4.89 (s, 1 H), 4.83 - 4.82 (m, 1 H), 4.77 (d, J = 11.2 Hz, 1 H), 4.47 - 4.37 (m, 1 H), 4.26 - 4.12 (m, 4 H), 3.82 - 3.42 (m, 5 H), 3.36 - 3.33 (m, 4 H), 3.27 (s, 1 H), 3.17 - 3.11 (m, 2 H), 3.02 (d, J = 14.4 Hz, 1 H), 2.97 - 2.91 (m, 3 H), 2.90 - 2.81 (m, 5 H), 2.74 - 2.60 (m, 3 H), 2.60 - 2.36 (m, 2 H), 2.29 - 2.12 (m, 2 H), 1.99 - 1.91 (m, 1 H), 1.88 - 1.75 (m, 1 H), 1.69 - 1.57 (m, 1 H), 1.42 (d, J = 6.0 Hz, 3 H), 1.17 - 0.88 (m, 10 H), 0.85 (d, J = 6.4 Hz, 3 H), 0.50 (s, 3 H) ppm.

[0190] Example 9 trans-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxamide TIFF2025522629000064.tif631704-The title compound was prepared in the same manner as in the preparation of Example 4, using trans 4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxylic acid (Compound 9g) instead of 4-(3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C). Example 9 (30.8 mg) was obtained as a white solid. MS calculated value 1116.5 (MH + ), measured value 1116.7 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.68 (d, J = 7.2 Hz, 1 H), 8.53 - 8.48 (m, 1 H), 7.70 (d, J = 2.4 Hz, 1 H), 7.51 - 7.45 (m, 2 H), 5.68 (t, J = 8.4 Hz, 1 H), 5.21 - 5.14 (m, 1 H), 4.80 (d, J = 10.8 Hz, 1 H), 4.46 - 4.37 (m, 1 H), 4.25 - 4.18 (m, 2 H), 4.15 - 3.87 (m, 2 H), 3.81 - 3.76 (m, 1 H), 3.73 - 3.64 (m, 2 H), 3.45 (d, J = 14.8 Hz, 1 H), 3.35 (s, 3 H), 3.27 - 3.23 (m, 1 H), 3.16 (s, 1 H), 3.09 (s, 3 H), 3.00 (s, 3 H), 2.88 - 2.76 (m, 2 H), 2.61 - 2.53 (m, 2 H), 2.26 - 1.84 (m, 12 H), 1.66 - 1.56 (m, 4 H), 1.50 - 1.43 (m, 6 H), 1.02 - 0.83 (m, 9 H), 0.44 (s, 3 H) ppm.

[0191] 9 g of this compound was prepared according to the following scheme. TIFF2025522629000065.tif102170

[0192] Step 1: Preparation of trans 4-tert-butyl 1-methylcyclohexane-1,4-dicarboxylate (Compound 9b) To a solution of trans 4-methoxycarbonylcyclohexanecarboxylic acid (Compound 9a, 5.0 g, 26.85 mmol) in tert-butanol (100 mL) was added 4-dimethylaminopyridine (6.6 g, 53.7 mmol), and then di-t-butyl dicarbonate (6.5 g, 29.54 mmol) was slowly added to the reaction mixture at 20 °C. The reaction mixture was stirred at 20 °C for 2 hours. After completion of the reaction, H2O (500 mL) was added to the reaction mixture, and the mixture was extracted with EtOAc (80 mL, 3 times). The combined organic layers were washed with brine (400 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by column chromatography (EtOAc in PE = 0% - 10%) to obtain trans 4-tert-butyl 1-methylcyclohexane-1,4-dicarboxylate (Compound 9b, 6.5 g) as a white solid. 1 H NMR (400 MHz, CHLOROFORM-d) δ = 3.67 (s, 3 H), 2.32 - 2.23 (m, 1 H), 2.21 - 2.12 (m, 1 H), 2.05 - 1.96 (m, 4 H), 1.44 - 1.42 (m, 13 H) ppm.

[0193] Step 2: Preparation of trans tert-butyl 4-(hydroxymethyl)cyclohexanecarboxylate (Compound 9c) To a solution of trans 4-tert-butyl 1-methylcyclohexane-1,4-dicarboxylate (Compound 9b, 5.0 g, 20.63 mmol) in THF (100 mL) was slowly added LiBH4 (1.4 g, 64.28 mmol) at 0 °C. The reaction mixture was stirred at 20 °C for an additional 12 h. After the reaction was completed, it was slowly quenched with H2O (500 mL) at 0 °C, and the reaction mixture was extracted with EtOAc (100 mL, 3 times). The combined organic layers were washed with brine (600 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by column chromatography (EtOAc in PE = 10% - 30%) to afford trans tert-butyl 4-(hydroxymethyl)cyclohexanecarboxylate (Compound 9c, 3.8 g) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d) δ: 3.47 (d, J = 6.0 Hz, 2 H), 2.18 - 2.08 (m, 1 H), 2.03 - 1.94 (m, 2 H), 1.91 - 1.79 (m, 2 H), 1.58 - 1.26 (m, 14 H) ppm.

[0194] Step 3: Preparation of trans tert-butyl 4-formylcyclohexanecarboxylate (Compound 9d) To a solution of trans tert-butyl 4-(hydroxymethyl)cyclohexanecarboxylate (Compound 9c, 3.8 g, 17.73 mmol) in DCM (100 mL) was slowly added DMP (11.28 g, 26.6 mmol) at 0 °C. The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated under vacuum to directly remove DCM, giving a residue. The residue was purified by column chromatography (EtOAc in PE = 10% - 30%) and concentrated under vacuum to afford trans tert-butyl 4-formylcyclohexanecarboxylate (Compound 9d, 2.5 g) as a yellow oil. 11H NMR (400 MHz, CHLOROFORM-d) δ = 9.63 (d, J = 1.2 Hz, 1 H), 2.33 - 1.93 (m, 6 H), 1.59 - 1.20 (m, 13 H) ppm.

[0195] Step 4: Preparation of trans tert-butyl 4-ethynylcyclohexanecarboxylate (Compound 9e) To a solution of trans tert-butyl 4-formylcyclohexanecarboxylate (Compound 9d, 2.5 g, 11.78 mmol) in methanol (50 mL) was added potassium carbonate (3.5 g, 25.32 mmol). The reaction mixture was cooled to 0 °C, and dimethyl (1-diazo-2-oxopropyl)phosphonate (3.5 g, 18.22 mmol) was slowly added. The reaction mixture was stirred at 20 °C for an additional 3 h. After completion of the reaction, H2O (200 mL) was added to the reaction mixture, which was then extracted with PE (60 mL, 2 times). The combined organic layers were washed with brine (800 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by column chromatography (EtOAc in PE = 0% - 16%) to afford trans tert-butyl 4-ethynylcyclohexanecarboxylate (Compound 9e, 1.5 g) as a white solid. 1 1H NMR (400 MHz, CHLOROFORM-d) δ = 2.29 - 1.94 (m, 7 H), 1.49 - 1.34 (s, 13 H) ppm.

[0196] Step 5: Preparation of trans tert-butyl 4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxylate (Compound 9f) To a solution of TMEDA (450 mg, 3.87 mmol) in DMF (15 mL) were added CuI (740 mg, 3.6 mmol) and potassium carbonate (1 g, 7.24 mmol) at 25 °C. After stirring vigorously for 20 minutes at 25 °C, TMSCF3 (700 mg, 4.92 mmol) was added to the reaction mixture under an argon atmosphere, and the mixture was stirred for an additional 20 minutes at 25 °C. The reaction mixture was cooled to 0 °C, and a mixture of trans tert-butyl 4-ethynylcyclohexanecarboxylate (Compound 9e, 500.0 mg, 2.4 mmol) and TMSCF3 (700 mg, 4.92 mmol) in DMF (10 mL) was added. The reaction mixture was stirred at 0 °C for 30 minutes and warmed to 25 °C for 12 hours. After completion of the reaction, the reaction mixture was poured into water (60 mL) and extracted with ethyl acetate (50 mL, 3 times). The combined organic layers were washed with brine (80 mL, 3 times), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue, which was purified by column chromatography (EtOAc in PE: 0% - 10%) to give trans tert-butyl 4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxylate (Compound 9f, 500 mg, 1.48 mmol) as a white solid. 1 H NMR (400 MHz, CHLOROFORM-d) δ = 2.17 - 1.94 (m, 6 H), 1.48 - 1.34 (m, 13 H) ppm.

[0197] Step 6: Preparation of trans 4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxylic acid (Compound 9g) To a solution of trans tert-butyl 4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxylate (Compound 9f, 100.0 mg, 0.36 mmol) in DCM (1 mL) was added TFA (1.0 mL). The reaction mixture was stirred at 20 °C for 0.5 hour. After completion of the reaction, the reaction mixture was concentrated under vacuum to obtain a residue. The residue was co-evaporated with DCM (5 mL) three times to give trans 4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxylic acid (Compound 9g, 50.0 mg) as a white solid, which was used directly in the next step without purification.

[0198] Example 10 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxamide TIFF2025522629000066.tif631704-(3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C) was replaced with cis 4-(3,3,3-trifluoroprop-1-ynyl)cyclohexanecarboxylic acid (Compound 10g), and the title compound was prepared in the same manner as in the preparation of Example 4. Example 10 (39.7 mg) was obtained as a white solid. MS calculated value 1116.5 (MH + ), measured value 1116.4 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 8.74 - 8.65 (m, 1H), 8.52 - 8.49 (m, 1H), 7.72 - 7.64 (m, 1H), 7.53 - 7.42 (m, 2H), 5.75 - 5.61 (m, 1H), 5.26 - 5.12 (m, 1H), 4.99 - 4.92 (m, 1H), 4.50 - 4.35 (m, 1H), 4.28 - 4.17 (m, 2H), 4.17 - 3.88 (m, 2H), 3.82 - 3.74 (m, 1H), 3.73 - 3.51 (m, 3H), 3.48 - 3.43 (m, 1H), 3.37 - 3.34 (m, 3H), 3.29 - 3.25 (m, 1H), 3.17 - 3.11 (m, 1H), 3.09 - 3.03 (m, 3H), 3.01 - 2.97 (m, 3H), 2.90 - 2.78 (m, 2H), 2.57 (d, J = 14.8 Hz, 1H), 2.45 - 2.31 (m, 1H), 2.29 - 2.11 (m, 2H), 2.05 - 1.89 (m, 4H), 1.88 - 1.71 (m, 10H), 1.70 - 1.57 (m, 2H), 1.45 (d, J = 6.0 Hz, 3H), 1.14 - 0.91 (m, 6H), 0.86 (d, J = 6.8 Hz, 3H), 0.44 (s, 3H) ppm.

[0199] Compound 10g was prepared in the same manner as Compound 9g, using cis 4-methoxycarbonylcyclohexanecarboxylic acid (Compound 10a) instead of trans 4-methoxycarbonylcyclohexanecarboxylic acid (Compound 9a).

[0200] Example 11 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide TIFF2025522629000067.tif62170trans-(2S)-3-Methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C) instead of cis-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 3B) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Using octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate G), the title compound was prepared in the same manner as in the preparation of Example 1. Example 11 (39.6 mg) was obtained as a yellow solid. MS calculated value 1021.5 (MH + ), measured value 1021.5 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.71 (d, J = 7.6 Hz, 1H), 8.39 (d, J = 2.8 Hz, 1H), 7.88 - 7.82 (m, 1H), 7.62 (d, J = 2.4 Hz, 1H), 7.40 - 7.32 (m, 1H), 5.84 - 5.72 (m, 1H), 4.84 - 4.72 (m, 1H), 4.47 - 4.34 (m, 2H), 4.32 - 4.22 (m, 1H), 4.20 - 4.12 (m, 1H), 4.10 - 3.98 (m, 1H), 3.87 (t, J = 4.8 Hz, 4H), 3.81 - 3.69 (m, 2H), 3.53 - 3.36 (m, 9H), 3.29 - 3.18 (m, 2H), 3.08 - 2.97 (m, 1H), 2.95 - 2.87 (m, 3H), 2.83 - 2.58 (m, 4H), 2.55 - 2.36 (m, 2H), 2.27 - 2.12 (m, 2H), 2.00 - 1.91 (m, 1H), 1.86 - 1.74 (m, 1H), 1.71 - 1.56 (m, 1H), 1.46 (d, J = 6.4 Hz, 3H), 1.07 - 0.94 (m, 9H), 0.89 - 0.82 (m, 3H), 0.68 - 0.49 (m, 3H) ppm.

[0201] Example 12 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide TIFF2025522629000068.tif65170trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C) was replaced by cis-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 3B) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Using octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate H), the title compound was prepared in the same manner as in the preparation of Example 1. Example 12 (7.7 mg) was obtained as a white solid. MS calculated value 1088.5 (MH + ), measured value 1088.5 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.54 - 8.50 (m, 1H), 7.54 - 7.42 (m, 4H), 5.84 - 5.96 (m, 1H), 5.20 - 5.10 (m, 1H), 4.68 - 4.60 (m, 1H), 4.39 (d, J = 12 Hz, 1H), 4.12 - 3.96 (m, 2H), 3.75 - 3.60 (m, 3H), 3.57 - 3.46 (m, 2H), 3.45 - 3.38 (m, 1H), 3.29 - 3.23 (m, 5H), 3.22 - 3.12 (m, 5H), 3.03 - 3.07 (m, 1H), 3.00 (s, 3H), 2.98 - 2.95 (m, 3H), 2.93 - 2.75 (m, 2H), 2.74 - 2.53 (m, 4H), 2.50 - 2.34 (m, 2H), 2.25 - 2.16 (m, 1H), 1.71 - 1.59 (m, 1H), 1.47 - 1.43 (m, 3H), 1.38 - 1.14 (m, 3H), 0.98 - 0.92 (m, 3H), 0.85 - 0.78 (m, 6H), 0.76 - 0.68 (m, 1H), 0.65 - 0.55 (m, 2H) ppm.

[0202] Example 13 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 [[Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[5-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutanecarboxamide TIFF2025522629000069.tif581704 - The title compound was prepared in the same manner as in the preparation of Example 4, using cis-3-[2-[5-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutane carboxylic acid (Compound 13B) instead of (3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C). Example 13 (10.8 mg) was obtained as a white solid. MS calculated value 1166.5 (MH + ), measured value 1166.4 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 9.07 (s, 2H), 8.66 (d, J = 7.3 Hz, 1H), 8.49 (d, J = 2.9 Hz, 1H), 7.67 (s, 1H), 7.46 - 7.38 (m, 2H), 5.65 - 5.61 (m, 1H), 5.15 (br d, J = 7.8 Hz, 1H), 4.68 - 4.56 (m, 1H), 4.46 - 4.37 (m, 1H), 4.26 - 4.19 (m, 2H), 3.80 - 3.67 (m, 3H), 3.61 (q, J = 7.2 Hz, 5H), 3.48 - 3.39 (m, 6H), 3.15 (br d, J = 14.7 Hz, 1H), 2.96 (d, J = 7.3 Hz, 6H), 2.83 - 2.76 (m, 2H), 2.70 - 2.66 (m, 1H), 2.60 - 2.52 (m, 2H), 2.22 (dt, J = 3.2, 7.5 Hz, 2H), 1.99 - 1.93 (m, 1H), 1.81 (br s, 1H), 1.68 - 1.60 (m, 1H), 1.45 - 1.41 (m, 3H), 1.32 - 1.27 (m, 1H), 1.18 (t, J = 6.8 Hz, 4H), 1.01 - 0.93 (m, 6H), 0.87 (d, J = 6.4 Hz, 3H), 0.45 - 0.40 (s, 3H) ppm.

[0203] Compound 13B was prepared in the same manner as Compound 5B using cis-tert-butyl 3-ethynylcyclobutanecarboxy (Intermediate I) and 2-iodo-5-(trifluoromethyl)-pyrimidine instead of cis-tert-butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1G) and 2-iodopyrimidine.

[0204] Example 14 N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide TIFF2025522629000070.tif58170trans-(2S)-3-Methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C) instead of (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E) 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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Using octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate D), the title compound was prepared in the same manner as in the preparation of Example 1. Example 14 (41.2 mg) was obtained as a yellow solid. MS calculated value 1035.5 (MH + ), measured value 1035.5 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.66 (d, J = 7.6 Hz, 1H), 8.48 (d, J = 2.8 Hz, 1H), 7.65 (d, J = 2.4 Hz, 1H), 7.50 (d, J = 2.8 Hz, 1H), 7.33 (d, J = 12.8 Hz, 1H), 5.79 - 5.74 (m, 1H), 4.46 - 4.13 (m, 12H), 4.08 - 4.03 (m, 2H), 3.77 - 3.63 (m, 6H), 3.49 - 3.46 (m, 1H), 3.45 - 3.43 (m, 1H), 3.09 - 3.01 (m, 2H), 3.00 (s, 3H), 2.95 (d, J = 2.4 Hz, 2H), 2.85 (s, 3H), 2.81 - 2.74 (m, 1H), 2.64 - 2.57 (m, 1H), 2.32 - 2.14 (m, 3H), 1.99 - 1.92 (m, 1H), 1.86 - 1.76 (m, 1H), 1.68 - 1.60 (m, 1H), 1.43 (d, J = 6.4 Hz, 3H), 0.98 - 0.92 (m, 12H), 0.50 (s, 3H).

[0205] Example 15 N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide TIFF2025522629000071.tif58170trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Instead of octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C), (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Using octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate F), the title compound was prepared in the same manner as in the preparation of Example 1. Example 15 (43.2 mg) was obtained as a yellow solid. MS calculated value 1103.5 (MH + ), measured value 1103.5 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 8.70 (d, J = 7.2 Hz, 1H), 8.39 - 8.36 (m, 1H), 7.80 - 7.76 (m, 1H), 7.68 - 7.62 (m, 1H), 7.38 - 7.32 (m, 1H), 5.85 - 5.78 (m, 1H), 4.40 - 4.14 (m, 8H), 4.10 - 4.02 (m, 2H), 3.78 - 3.68 (m, 3H), 3.49 - 3.45 (m, 4H), 3.45 - 3.42 (s, 1H), 3.19 - 3.14 (m, 2H), 2.94 - 2.85 (m, 6H), 2.83 (s, 4H), 2.75 - 2.69 (m, 1H), 2.22 - 2.14 (m, 2H), 2.01 - 1.92 (m, 1H), 1.86 - 1.75 (m, 1H), 1.69 - 1.57 (m, 1H), 1.45 (d, J = 6.4 Hz, 3H), 1.34 - 1.23 (m, 1H), 1.05 - 0.99 (m, 3H), 0.97 - 0.90 (m, 11H), 0.58 (s, 3H).

[0206] Example 16 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[4-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutanecarboxamide Instead of TIFF2025522629000072.tif551704-(3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C), cis-3-[2-[4-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutanecarboxylic acid (Compound 16B) was used, and the title compound was prepared in the same manner as in the preparation of Example 4. Example 16 (98.2 mg) was obtained as a white solid. MS calculated value 1166.5 (MH + ), measured value 1166.4 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 9.06 - 8.99 (m, 1H), 8.71 - 8.63 (m, 1H), 8.47 - 8.39 (m, 1H), 7.82 - 7.73 (m, 1H), 7.71 - 7.64 (m, 1H), 7.46 - 7.26 (m, 2H), 5.77 - 5.60 (m, 1H), 5.20 - 5.02 (m, 3H), 4.48 - 4.09 (m, 2H), 3.82 - 3.74 (m, 1H), 3.72 - 3.66 (m, 1H), 3.60 - 3.53 (m, 1H), 3.52 - 3.46 (m, 1H), 3.46 - 3.42 (m, 1H), 3.41 - 3.38 (m, 1H), 3.38 - 3.34 (m, 4H), 3.17 - 3.08 (m, 1H), 3.03 - 2.92 (m, 3H), 2.91 - 2.84 (m, 1H), 2.83 - 2.77 (m, 1H), 2.76 - 2.69 (m, 2H), 2.66 - 2.60 (m, 5H), 2.60 - 2.54 (m, 2H), 2.36 (s, 3H), 2.29 - 2.15 (m, 2H), 2.01 - 1.90 (m, 1H), 1.86 - 1.70 (m, 1H), 1.69 - 1.53 (m, 1H), 1.46 - 1.38 (m, 3H), 1.32 - 1.25 (m, 1H), 1.20 - 1.02 (m, 1H), 1.00 - 0.94 (m, 5H), 0.92 - 0.82(m, 4H), 0.81 - 0.69 (m, 1H), 0.40 (s, 3H).

[0207] cis-tert-Butyl (2S)-2-[(3-ethynylcyclobutanecarbonyl)-methyl-amino]-3-methyl-butanoate (Compound 1G) and 2-bromopyrimidine were replaced with cis-tert-butyl 3-ethynylcyclobutanecarboxy (Intermediate I) and 2-bromo-4-(trifluoromethyl)pyrimidine, and Compound 16B was prepared in the same manner as Compound 5B.

[0208] Example 18 N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide TIFF2025522629000073.tif64170trans-(2S)-3-Methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26(2S)-3-Methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 (Octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E)) was used to prepare the title compound in the same manner as in the preparation of Example 1. Example 18 (24.8 mg) was obtained as a yellow solid. MS calculated value 1157.5 (MH + ), measured value 1157.5 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 8.71 (d, J = 7.2 Hz, 1H), 8.41 (d, J = 2.8 Hz, 1H), 7.73 - 7.64 (m, 2H), 7.48 (d, J = 12.4 Hz, 1H), 5.79 - 5.72 (m, 1H), 5.23 - 5.16 (m, 1H), 4.44 - 4.28 (m, 4H), 4.25 - 3.99 (m, 4H), 3.83 - 3.73 (m, 1H), 3.72 - 3.69 (m, 1H), 3.50 - 3.42 (m, 5H), 3.40 - 3.36 (s, 3H), 3.25 - 3.14 (m, 3H), 2.91 - 2.82 (m, 9H), 2.66 (d, J = 14.4 Hz, 1H), 2.24 - 2.16 (m, 2H), 1.99 - 1.91 (m, 1H), 1.91 - 1.68 (m, 1H), 1.68 - 1.55 (m, 1H), 1.48 - 1.45 (m, 3H), 0.99 - 0.91 (m, 11H), 0.51 (s, 3H).

[0209] Example 19 (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-Tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-Hexaene-7-yl]-2-Isopropyl-4-Oxo-4-[3-(3,3,3-Trifluoroprop-1-ynyl)azetidin-1-yl]butanamide Instead of TIFF2025522629000074.tif54170BOC-N-ME-VAL-OH and 4-(3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C), (2S)-4-tert-butoxy-2-isopropyl-4-oxo-butanoic acid and 3-(3,3,3-trifluoroprop-1-ynyl)azetidine (Compound 19B) were used to prepare the title compound in the same manner as the preparation of Example 4. Example 19 (30.6 mg) was obtained as a white solid. MS calculated value 1074.4 (MH + ), measured value 1074.4 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.67 (d, J = 7.6 Hz, 1H), 8.50 (d, J = 2.8 Hz 1H), 7.76 - 7.68 (m, 1H), 7.55 (s, 1H), 7.50 - 7.44 (m, 1H), 5.78 - 5.66 (m, 1H), 5.22 - 5.13 (m, 1H), 4.98 - 4.89 (m, 2H), 4.57 - 4.48 (m, 1H), 4.46 - 4.39 (m, 1H), 4.35 - 4.27 (m, 1H), 4.26 - 4.16 (m, 3H), 4.14 - 4.04 (m, 1H), 4.04 - 3.99 (m, 1H), 3.98 - 3.92 (m, 1H), 3.82 - 3.76 (m, 1H), 3.74 - 3.62 (m, 3H), 3.59 - 3.52 (m, 1H), 3.50 - 3.39 (m, 2H), 3.38 - 3.33 (m, 4H), 3.21 - 3.09 (m, 2H), 3.00 (s, 3H), 2.86 - 2.70 (m, 2H), 2.64 - 2.44 (m, 2H), 2.26 - 2.14 (m, 2H), 2.00 - 1.88 (m, 2H), 1.86 - 1.73 (m, 1H), 1.70 - 1.58 (m, 1H), 1.45 (d, J = 6.0 Hz, 3H), 1.26 - 1.06 (m, 1H), 1.03 - 0.92 (m, 9H), 0.53 - 0.41 (m, 3H).

[0210] Compound 19B was prepared in the same manner as Compound 9g, using tert-butyl 3-ethynylazetidine-1-carboxylate (Compound 2A) instead of trans tert-butyl 4-ethynylcyclohexanecarboxylate (Compound 9e).

[0211] Example 20 N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carboxamide TIFF2025522629000075.tif57170trans-(2S)-3-Methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26(2S)-3-Methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was used to prepare the title compound in the same manner as in the preparation of Example 1. Example 20 (8.8 mg) was obtained as a yellow solid. MS calculated value 1022.5 (MH + ), measured value 1022.5 (MH + ). 11H NMR (400 MHz, DMSO-d6) δ = 8.55 (d, J = 7.6 Hz, 1H), 8.50 - 8.39 (m, 2H), 7.67 (d, J = 2.4 Hz, 1H), 7.56 (d, J = 12.8 Hz, 1H), 7.30 (d, J = 1.6 Hz, 1H), 5.53 - 5.41 (m, 1H), 5.14 (d, J = 12.0 Hz, 1H), 4.31 - 4.21 (m, 4H), 4.21 - 4.05 (m, 5H), 4.05 - 3.98 (m, 2H), 3.81 - 3.70 (m, 6H), 3.57 (s, 2H), 3.26 - 3.23 (m, 3H), 3.20 (s, 3H), 2.93 - 2.84 (m, 1H), 2.78 - 2.68 (m, 4H), 2.11 - 2.03 (m, 2H), 1.88 - 1.67 (m, 2H), 1.58 - 1.45 (m, 1H), 1.33 (d, J = 6.0 Hz, 3H), 1.23 (s, 1H), 0.95 - 0.83 (m, 10H), 0.79 (d, J = 6.8 Hz, 3H), 0.37 (s, 3H).

[0212] Example 21 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[4-(trifluoromethyl)pyrimidin-2-yl]ethynyl]cyclobutanecarboxamide TIFF2025522629000076.tif551704 - (3,3,3-trifluoroprop-1-ynyl)benzoic acid (Compound 4C) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C) was replaced with cis-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 16B) and (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-tetrazapentacyclo-[17.5.2.1 2,5 .1 9,13 .0 22,26 octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was used to prepare the title compound in the same manner as in the preparation of Example 4. Example 21 (22.8 mg) was obtained as a white solid. MS calculated value 1099.5 (MH + ), measured value 1099.5 (MH + ). 11H NMR (400 MHz, Methanol-d4) δ = 9.03 (d, J = 5.2 Hz, 1H), 8.66 - 8.61 (m, 1H), 8.40 (d, J = 2.8 Hz, 1H), 7.78 (d, J = 5.0 Hz, 1H), 7.62 (d, J = 2.4 Hz, 1H), 7.32 (d, J = 2.8 Hz, 1H), 7.25 (d, J = 12.8 Hz, 1H), 5.71 (br d, J = 9.0 Hz, 1H), 4.92 - 4.88 (m, 1H), 4.80 - 4.76 (m, 1H), 4.47 - 4.38 (m, 1H), 4.27 - 4.22 (m, 1H), 4.21 - 4.16 (m, 2H), 3.88 - 3.84 (m, 4H), 3.78 - 3.69 (m, 2H), 3.56 - 3.48 (m, 1H), 3.46 - 3.36 (m, 2H), 3.28 - 3.26 (m, 3H), 3.05 - 2.92 (m, 5H), 2.86 - 2.52 (m, 8H), 2.25 - 2.17 (m, 2H), 1.99 - 1.92 (m, 1H), 1.88 - 1.74 (m, 1H), 1.68 - 1.57 (m, 1H), 1.42 (d, J = 6.4 Hz, 3H), 1.34 - 1.28 (m, 1H), 1.12 - 1.02 (m, 1H), 0.99 - 0.84 (m, 12H), 0.53 - 0.45 (m, 3H).

[0213] Example 22 cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarboxamide TIFF2025522629000077.tif55170trans-(2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)cyclobutanecarbonyl]amino]butanoic acid (Compound 1I) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Instead of octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate C), (2S)-3-methyl-2-[methyl-[3-(3,3,3-trifluoroprop-1-ynyl)azetidine-1-carbonyl]amino]butanoic acid (Compound 2E) and (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-tetrazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 Using octacosa-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (Intermediate K), the title compound was prepared in the same manner as in the preparation of Example 1. Example 22 (41.5 mg) was obtained as a white solid. MS calculated value 1075.4 (MH + ), measured value 1075.4 (MH + ). 11H NMR (400 MHz, methanol-d4) δ = 8.80 - 8.63 (d, J = 7.2 Hz, 1H), 8.44 - 8.36 (d, J = 2.8 Hz, 1H), 7.71 - 7.61 (m, 2H), 7.53 - 7.42 (d, J = 12.8 Hz, 1H), 5.76 - 5.63 (m, 1H), 5.27 - 5.11 (m, 2H), 4.80 - 4.72 (m, 1H), 4.49 - 4.36 (m, 1H), 4.34 - 4.13 (m, 2H), 3.94 - 3.84 (m, 4H), 3.83 - 3.76 (m, 1H), 3.75 - 3.67 (m, 1H), 3.58 - 3.43 (m, 2H), 3.39 - 3.35 (m, 6H), 3.29 - 3.23 (m, 1H), 3.17 - 3.09 (m, 1H), 2.98 - 2.90 (m, 3H), 2.88 - 2.77 (m, 1H), 2.75 - 2.60 (m, 3H), 2.56 - 2.43 (m, 2H), 2.30 - 2.15 (m, 2H), 2.04 - 1.91 (m, 1H), 1.90 - 1.74 (m, 1H), 1.72 - 1.56 (m, 1H), 1.51 - 1.43 (d, J = 6.0 Hz, 3H), 1.41 - 1.34 (d, J = 6.4 Hz, 1H), 1.22 - 1.14 (d, J = 6.4 Hz, 1H), 1.09 - 1.03 (d, J = 6.4 Hz, 1H), 1.01 - 0.95 (m, 5H), 0.92 - 0.80 (m, 3H), 0.55 - 0.45 (m, 3H).

[0214] Biological Examples Compound A168 on page 81 of Table 1 in International Publication No. 2021091982 was cited as the reference compound of the present invention. TIFF2025522629000078.tif69170

[0215] Example 23 GSH reaction rate Glutathione (GSH) is a tripeptide found in most tissues, particularly at high concentrations in the liver, and plays an important role in protecting cells from oxidative damage and toxicity by xenobiotic electrophiles and in maintaining redox homeostasis. More specifically, glutathione conjugation contributes to detoxification by binding to electrophiles that would otherwise bind to proteins or nucleic acids and cause cell damage and genetic mutations.

[0216] Many potentially toxic electrophilic xenobiotics and some endogenous compounds are detoxified by conversion to their corresponding glutathione S-conjugates, which consumes the endogenous GSH and subsequently reduces the detoxification capacity. Some drugs and halogenated workplace / environmental pollutants are bioactivated by this mechanism.

[0217] On the other hand, conjugation between glutathione and drug molecules in extrahepatic organs as well as in the liver typically results in poor PK properties of the molecule (especially high clearance) and increases the potential for its off-target reactivity (a potential cause of various toxicities). Therefore, strategies to minimize GSH metabolism are highly important. The short T in the native GSH reaction 1 / 2 showed a high GSH reaction rate. Therefore, the T in the native GSH reaction assay 1 / 2 was determined for candidate screening.

[0218] The reference compound and the compounds of the present invention may form conjugates with GSH by either a halogenated moiety substitution reaction or a direct Michael addition reaction. Therefore, this test was performed to confirm the GSH reactivity of the listed compounds.

[0219] For the determination of the inherent GSH reactivity, 1 μM of the compound was incubated at 37 °C for 0, 0.5, 1, 2, 4, and 6 h in 100 mM potassium phosphate buffer at pH 7.4 with and without 5 mM GSH. At the end of the specified time points, the samples were quenched with acetonitrile containing 10 mM N-ethylmaleimide and an internal standard. The quenched samples were centrifuged and the supernatant was analyzed by LC-MS / MS for compound quantification. If the depletion % after 6 h of incubation was less than 20%, the compound was reported to be stable, and if the depletion % was greater than 20%, the half-life value was reported. TIFF2025522629000079.tif60170

[0220] The above results clearly show that the reference compound (A168) formed a conjugate with GSH and caused its depletion over 6 h, while the compounds of the present invention maintained stability with much less or no conjugation with GSH.

[0221] Example 24 Single-dose pharmacokinetics (SDPK) study in female BALB / c mice The purpose of this study was to determine the pharmacokinetics of the selected compounds after single intravenous bolus or oral gavage administration in female BALB / c mice. Briefly, two groups of female BALB / c mice (available from Zhejiang Vital River Laboratory Animal Technology Co., Ltd. or Shanghai Lingchang Biotechnology Co., Ltd.) (N = 3 / group) were administered 3 mg / kg of the compound intravenously (IV) or 30 mg / kg of the compound orally (PO). Blood samples were collected at 5 min (only for IV), 15 min, 30 min, 1 h, 2 h, 4 h, 7 h, and 24 h after administration. The blood samples were placed on ice until centrifuged to obtain plasma samples. The compound concentration in the plasma samples was measured using the LC-MS / MS method. Pharmacokinetic parameters were calculated by non-compartmental analysis. TIFF2025522629000080.tif51170

[0222] From Table 2, it can be seen that Example 11 has good pharmacokinetic properties in the mouse model. In particular, compared with A168, Example 11 has C max almost doubled 、 AUC 0-last is 1.5 times, and the clearance is very low. Therefore, Example 11 is more suitable for the treatment of cancers with KRAS mutations as an orally administered therapeutic active ingredient in clinical practice.

[0223] Example 25 Human hepatocyte stability assay The hepatocyte stability assay measures the disappearance rate of a compound from incubation with cryopreserved suspension hepatocytes derived from humans. Positive controls including midazolam, raloxifene, and dextromethorphan are included in all experiments. The incubation consists of a 1 μM test compound and human hepatocytes (1×10 6 cells / mL) suspension in William’s E medium supplemented with 10% FBS and 0.5% penicillin-streptomycin. The hepatocyte suspension was incubated in a 5% CO2 incubator at 37 °C with intermittent shaking at 900 rpm. The reaction was stopped by adding methanol containing an internal standard (2 μM tolbutamide) at 2, 10, 20, 40, 60, and 120 minutes after compound addition, and the depletion of the parent compound was monitored by LC-MS / MS analysis. For human data, CL_hep (mL / min / kg) > 16.24 represents high clearance, CL_hep (mL / min / kg) < 6.96 represents low clearance, and 16.24 < CL_hep (mL / min / kg) > 6.96 represents moderate clearance. TIFF2025522629000081.tif67170

[0224] Achieving low clearance is advantageous for improving the in vivo performance of the present compound, such as reducing dosage, increasing exposure, and extending the half-life. The above results clearly show that in the human hepatocyte stability assay, the reference compound (A168) showed moderate clearance, while the compounds of the present invention maintained low clearance.

[0225] Example 26 Cell viability assay The purpose of this cell assay was to determine the effect of test compounds 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 Cell Counting Kit-8.

[0226] Cells were seeded at 5,000 cells / well (NCI-H358), 2,000 cells / well (AGS), and 2,000 cells / well (SW620) in a 96-well assay plate (Corning-3699) and incubated overnight. Then, on the day of the assay, the 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 (value obtained by subtracting OD650 from OD450) was read using EnVision. IC 50 was determined by fitting a 4-parameter sigmoid concentration-response model. TIFF2025522629000082.tif212170

[0227] Example 27 KRAS G12C - BRAF NanoBit assay This assay is for measuring the ability of test compounds to disrupt the KRAS G12C - BRAF complex at the cellular level, and the inventors established a NanoBit cell assay in mammalian HEK293 (ATCC) cells.

[0228] HEK293 cells were grown and maintained using DMEM medium (Thermo Fisher Scientific) containing 10% fetal bovine serum and 1% penicillin / streptomycin. Both KRAS G12C and BRAF RBD were cloned into the NanoBit vector (BiBiT vector system, Promega) in the directions of SmBit-KRAS G12C and BRAF RBD-LgBit respectively, and co-transfected into HEK293 cells. Then, the cells were selected for 4 weeks using 100 μg / mL hygromycin B (10687010, Thermo Fisher) and blasticidin (5 μg / mL) to obtain a stable cell pool.

[0229] On the day of the assay, 75 nL of the compound solution was present in the 384-well assay plate as a 16-point 3-fold dilution starting from a final concentration of 30 μM in DMSO. Then, the cells were seeded into the 384-well plate at 10,000 cells / 25 μL / well. After 3 hours of incubation, 6 μL of volume of Nano-Glo® Live Cell Substrate (Promega) was added to each well. Luminescence was monitored using the Envision ultra384 model in 20 minutes. Compounds that promoted the disruption of the KRAS G12C-BRAF RBD complex were identified as compounds that induced a decrease in luminescence compared to the DMSO control wells. TIFF2025522629000083.tif193170

[0230] Example 28 Interaction assay of CYPA (50 nM) with KRAS-BRAF 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 the binding of KRAS G12D or KRAS G12V to BRAF by the compounds of the present invention, respectively. In an assay buffer containing 25 mM HEPES pH = 7.4 (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, Thermo, 15630080), 0.002% Tween 20, 0.1% BSA, 100 mM NaCl, 5 mM MgCl2, 10 μM GMPPNP (guanosine 5'-[β,γ-imido]triphosphate trisodium salt hydrate, Sigma, G0635), tagless CYPA, GMPPNP-loaded 6His-KRAS protein, and GST-BRAF RBD were mixed in the wells of a 384-well assay plate at final concentrations of 50 nM, 6.25 nM, and 1 nM, respectively. The compounds were present in the plate wells as a 16-point 3-fold dilution series starting at a final concentration of 10 μM and incubated for 3 hours. Then, a mixture of MAb anti-6His-XL665 (Cisbio, 61HISXLB) and Mab anti-GST-TB cryptate (Cisbio, 61GSTTLB) was added at final concentrations of 6.67 nM and 0.21 nM, respectively, and the plate was incubated for an additional 1.5 hours. The TR-FRET signal was 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 compounds that induced a decrease in the TR-FRET ratio compared to DMSO control wells. TIFF2025522629000084.tif173170

[0231] Example 29 pERK inhibition assay This assay is for measuring the ability of test compounds to inhibit the phosphorylation of ERK; downstream signaling of KRAS G12C in NCI-H358 cells, KRAS G12D in AGS cells, and KRAS G12V in SW620 cells. All of NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL-1739) cells, and SW620 (ATCC-CCL-227) cells were grown and maintained using RPMI-1640 medium (Thermo Fisher Scientific) containing 10% fetal bovine serum and 1% penicillin / streptomycin. One day before compound addition, the cells were plated in tissue culture-treated 96-well plates (Corning-3699) at a density of 30,000 cells / well, 20,000 cells / well, and 30,000 cells / well for NCI-H358, AGS, and SW620, respectively, and allowed to adhere overnight. Then, the diluted compounds were added at a final concentration of 0.5% DMSO. Four hours after incubation, the medium was removed, 100 μL of 4% formaldehyde was added, and the assay plates were incubated at room temperature for 20 minutes. Next, the plates were washed once with phosphate-buffered saline (PBS) and permeabilized with 100 μL of cold methanol for 10 minutes. Nonspecific antibody binding to the plates was blocked with 50 μL of 1× BSA blocking buffer (10-fold dilution with phosphate-buffered saline Tween (PBST), Thermo-37520) at room temperature for at least 1 hour.

[0232] The amount of phospho-ERK was determined using an antibody specific for the phosphorylated form of ERK. The primary antibody (pERK, CST-4370, Cell Signaling Technology) was diluted 1:300 in blocking buffer, and 50 μL aliquots were added to each well and incubated overnight at 4 °C. Cells were washed five times for 5 minutes each with PBST. The secondary antibody (HRP-conjugated anti-rabbit IgG, CST-7074, Cell Signaling Technology) was diluted 1:1000 in blocking buffer, 50 μL was added to each well, and incubated at room temperature for 1-2 hours. Cells were washed five times for 5 minutes each with PBST, 100 μL of TMB ELISA substrate (abcam-ab171523) was added, and gently shaken for 20 minutes. 50 μL of stop solution (abcam-ab171529) was added, and then signals (OD450) were read by EnVision.

[0233] IC 50 was determined by fitting a 4-parameter sigmoid concentration-response model. TIFF2025522629000085.tif221170

[0234] Example 30 Stable KRAS mutant cell lines and cell survival assays. The purpose of this study was to determine the potency and efficacy of compounds against cell proliferation using the CellTiter-Glo® (CTG) Luminescent Cell Viability Assay (Promega, Madison, WI). The inventors identified 14 KRAS mutants with secondary mutations (V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L). G12CThe variant array was cloned into Miapaca-2. A total of 14 stable Miapaca-2 mutant cell lines were established by lentiviral infection. In the cell viability assay, a 4-fold dilution series was used with a maximum dose of 10 μM, and cells were dosed with the compound in a 9-point dose response. KRAS mutant cells were maintained in DMEM + 10% FBS + 2.5% HI horse serum + 1% PS + 1 μg / mL puromycin and seeded into 96-well plates at 800 - 1,500 cells per well 24 hours prior to compound addition and then incubated with the compound for 3 days before assaying viability (CellTiter-Glo, Promega). The assay was performed biologically in duplicate. A non-linear regression curve was fitted using Xfit. The IC50 (absolute IC50) is the dose at which the estimated viability is 50% relative to untreated wells. The inhibition rate of the compound is calculated according to the following formula: % inhibition = 100 - 100 × (luminescence value - HPE) / (ZPE - HPE). HPE: Luminescence value of wells with only medium added ZPE: Luminescence value of wells with DMSO added TIFF2025522629000086.tif46170

Claims

1. Compound of formula (I) [In the above formula, R １ teeth, And; {In the above formula, R ８ C １－６ It is alkyl, R ９ is cycloalkyl, azetidinyl or phenyl, and the cycloalkyl, azetidinyl and phenyl are substituted with halo C ３－７ alkynyl, (halo C ３－７ alkylpyrimidinyl)C ３－６ alkynyl or pyrimidinyl C ３－６ alkynyl} ２－６ ２－６ ​​ R ２ C １－６ It is alkyl; R ３ is H or a halogen; R ４ is H or a halogen; R ５ C １－６ Alkyl or Halo C １－６ It is alkyl; R ６ C １－６ Alkoxy C １－６ It is alkyl; R ７ is morpholinyl, (halo C １－６ Alkyl)piperazinyl or C １－６ It is alkylpiperazinyl; A １ It is thiazoylene; A ２ C １－６ It is alkylene; However, R ３ and R ４ [Not H at the same time] or a pharmaceutically acceptable salt thereof.

2. Compound of formula (Ia) [In the above formula, R １ teeth, And; {In the above formula, R ８ C １－６ It is alkyl, R ９ C ３－７ The C is a cycloalkyl, azetidinyl, or phenyl. ３－７ Cycloalkyl, azetidinyl, and phenyl are halo C ３－６ Alkinil, (Halo C ３－６ (Alkylpyrimidinyl) C ２－６ Alkinyl or pyrimidinyl C ２－６ Substituted with alkynyl} R ２ C １－６ It is alkyl; R ３ is H or a halogen; R ４ is H or a halogen; R ５ C １－６ Alkyl or Halo C １－６ It is alkyl; R ６ C １－６ Alkoxy C １－６ It is alkyl; R ７ is morpholinyl, (halo C １－６ Alkyl)piperazinyl or C １－６ It is alkylpiperazinyl; A １ It is thiazoylene; A ２ C １－６ It is alkylene; However, R ３ and R ４ [Not H at the same time] or a pharmaceutically acceptable salt thereof.

3. R １ but (In the formula, R ８ is C １－６ It is alkyl; R ９ Hello C ３－６ C substituted with alkynyl ３－７ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is a cycloalkyl compound.

4. R １ but (In the formula, R ８ is methyl; R ９ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein ( is cyclobutyl substituted with 3,3,3-trifluoropropa-1-inyl).

5. R ９ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is 3-(3,3,3-trifluoropropane-1-inyl)cyclobutyl.

6. R ２ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein isopropyl.

7. R ３ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the halogen is present.

8. R ３ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is fluoro.

9. R ４ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is H or fluoro.

10. R ４ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is H.

11. R ５ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is ethyl or 2,2,2-trifluoroethyl.

12. R ６ The compound according to claim 1, wherein is 1-methoxyethyl, or a pharmaceutically acceptable salt thereof.

13. R ７ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is morpholinyl, 4-(2,2,2-trifluoroethyl)piperazine-1-yl, or 4-methylpiperazine-1-yl.

14. A １ but The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the bond "a" is bonded to the indole ring.

15. A ２ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is dimethylmethylene.

16. R １ but (In the formula, R ８ is C １ - ６ It is alkyl, R ９ Hello C ３－６ C substituted with alkynyl ３－７ It is a cycloalkyl group; R ２ However, C １－６ It is alkyl; R ３ However, it is a halogen; R ４ However, it is H; R ５ is C １－６ alkyl or halo C １－６ alkyl; R ６ However, C １－６ Alkoxy C １－６ It is alkyl; R ７ However, morpholinyl, (halo C １－６ Alkyl)piperazinyl or C １－６ It is alkylpiperazinyl; A １ is (In the formula, bond "a" is bonded to the indole ring); A ２ However, C １－６ The compound according to claim 1, which is alkylene, or a pharmaceutically acceptable salt thereof.

17. R １ but, (In the formula, R ８ is methyl, R ９ (is 3-(3,3,3-trifluoropropane-1-inyl)cyclobutyl); R ２ However, it is isopropyl; R ３ However, it is fluoro; R ４ However, it is H; R ５ However, it is ethyl or 2,2,2-trifluoroethyl; R ６ However, it is (1S)-1-methoxyethyl; R ７ However, these are morpholinyl, 4-(2,2,2-trifluoroethyl)piperazine-1-yl, or 4-methylpiperazine-1-yl; A １ but, (In the formula, bond "a" is bonded to the indole ring); A ２ However, it is dimethylmethylene. The compound according to claim 16 or a pharmaceutically acceptable salt thereof.

18. The following compounds can be selected: trans-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide; N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)azetidine-1-carboxamide; cis-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide; N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoropropane-1-inyl)benzamide; cis-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(2-pyrimidine-2-ylethinyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazine-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazine-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide; trans-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoropropane-1-inyl)cyclohexanecarboxamide; cis-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(3,3,3-trifluoropropane-1-inyl)cyclohexanecarboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide; cis-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[5-(trifluoromethyl)pyrimidine-2-yl]ethinyl]cyclobutanecarboxamide; N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)azetidine-1-carboxamide; N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazine-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)azetidine-1-carboxamide; cis-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[4-(trifluoromethyl)pyrimidine-2-yl]ethinyl]cyclobutanecarboxamide; N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazine-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)azetidine-1-carboxamide; (2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazine-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-isopropyl-4-oxo-4-[3-(3,3,3-trifluoropropane-1-inyl)azetidine-1-yl]butanamide; N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)azetidine-1-carboxamide; cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-[4-(trifluoromethyl)pyrimidine-2-yl]ethinyl]cyclobutanecarboxamide; and cis-N-[(1S)-1-[[(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-tetrazapentacyclo[17.5.2.1 ２，５ 1. ９，１３ . 0 ２２，２６ Octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(3,3,3-trifluoropropane-1-inyl)cyclobutanecarboxamide or a pharmaceutically acceptable salt thereof.

19. A method for preparing a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, comprising the following steps: a) Compound of formula (II) in the presence of a coupling reagent and a base. and acid (III) The process includes a coupling reaction with to form a compound of formula (I), In the above formula, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ A １ and A ２ The coupling reagent is as defined in any one of claims 1 to 17; T ３ The base is P, HATU, PyBOP, or EDCI / HOBt; the base is TEA, DIEPA, or DMAP.

20. A compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 for use as a therapeutically active substance.

21. A pharmaceutical composition comprising a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable additive.

22. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof for the treatment of KRAS G12C protein-related disorders.

23. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof for the treatment of KRAS G12C, G12D, and G12V protein-related disorders.

24. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 for inhibiting RAS interaction with a downstream effector, wherein the downstream effector is RAF and PI3K.

25. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof for inhibiting the proliferation of oncogenic MAPK and PI3K signaling.

26. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 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.

27. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 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.

28. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 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.

29. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer comprises a first mutation which is G12C and a second mutation at a position selected from V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L.

30. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 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.

31. Use of a compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer comprises a first mutation which is G12C and a second mutation at a position selected from V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L.

32. A method for the treatment or prevention of KRAS mutation-driven cancer selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer, comprising administering a therapeutically effective amount of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof.

33. A method for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer comprises a first mutation which is G12C and a second mutation at a position selected from V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L.

34. A compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, as produced according to the method of claim 19.

35. The invention as previously described.

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