Macrocyclic compounds for treatment of cancer

Abstract

The present invention relates to compounds of formula (I) # imgabs0 # wherein R1 to R7, A1 and A2 are as described herein, and pharmaceutically acceptable salts thereof, as well as compositions comprising and methods of using the compounds.

Description

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

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

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

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

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

[0006] The first generation of KRASG12C inhibitors, such as Sotorosib and Adagrasib, which target the 'GDP bound off' form (RASOFF) of the KRAS G12C mutation, have shown good efficacy. Although this treatment has benefited many patients with activating KRAS mutations, almost all of the patients who initially benefit will eventually acquire resistance through various mechanisms. An increasing number of cases of KRAS G12C secondary mutations have been identified from patient samples, such as Y96D, R68S, H95D, H95Q, H95R, V8L (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 from saturation mutagenesis (Siyu et al., PNAS (2022)) and ENU mutagenesis (Takamasa et al., J Thorac Oncol (2021)), which show 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 novel compounds having formula (I),

[0008]

[0009] in

[0010] R 1 for 3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazinyl or (C 1-6 Alkyl) oxoimidazolidinyl;

[0011] Where R 8 C 1-6 alkyl;

[0012] R 9 for ((C 1-6 alkyl)2amino)azetidinyl, C 1-6 Alkylpiperazinyl, halogenated azetidinyl, halogenated C 1-6 Alkylamino, halogenated C 1-6 Alkylaminoazetidinyl, halo C 1-6 Alkylpiperazine, hydroxyl (C 1-6 alkyl)piperidinyl or morpholinyl;

[0013] R2 C 1-6 alkyl;

[0014] R 3 is H or halogen;

[0015] R 4 is H or halogen;

[0016] R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0017] R 6 C 1-6 Alkoxy C 1-6 alkyl;

[0018] R 7 For morpholinyl, (halogenated C 1-6 alkyl)piperazinyl or C 1-6 Alkylpiperazinyl;

[0019] A 1 is thiazolylidene;

[0020] A 2 C 1-6 Alkylene;

[0021] The prerequisite is R 3 and R 4 Not at the same time H;

[0022] or a pharmaceutically acceptable salt thereof.

[0023] The invention also relates to their manufacture, to medicaments based on the compounds according to the invention and their production and to the use of the compounds of formula (I) or (Ia) as KRAS inhibitors.

[0024] The compounds of formula (I) or (Ia) show good KRAS inhibition against G12C, G12D and G12V. In another embodiment, the compounds of the present invention show excellent cancer cell inhibition and human hepatocyte stability. In addition, the compounds of formula (I) or (Ia) also show good or improved cytotoxicity and solubility characteristics. In addition, compared with the reference compound, the compounds of the present invention have excellent pharmacokinetic properties. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 .X-ray crystallographic analysis of compound G5. DETAILED DESCRIPTION

[0026] definition

[0027] The term "C 1-6"Alkyl" means a saturated, straight-chain or branched alkyl group containing 1 to 6, especially 1 to 4 carbon atoms, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, etc. In particular, "C 1-6 "Alkyl" groups are methyl, ethyl and n-propyl.

[0028] The term "C 1-6 "Alkoxy" means C 1-6 Alkyl-O-.

[0029] The term “(C 1-6 ")Alkylene" means a straight or branched saturated divalent hydrocarbon group of 1 to 6 carbon atoms or a divalent branched saturated divalent hydrocarbon group of 3 to 6 carbon atoms. 1-6 ) Examples of the alkylene group include methylene, ethylene, propylene, 2-methylpropylene, butylene, 2-ethylbutylene, pentylene, hexylene.

[0030] The terms "halogen" and "halo" are used interchangeably herein to refer to fluorine, chlorine, bromine or iodine.

[0031] The term "halogenated C 1-6 "Alkyl" means C 1-6 Alkyl groups, where C 1-6 At least one hydrogen atom of the alkyl group has been replaced by identical or different halogen atoms, in particular fluorine atoms. Examples of haloalkyl groups include monofluoro-, difluoro- or trifluoro-methyl, -ethyl or -propyl, for example 3,3,3-trifluoropropyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, fluoromethyl or trifluoromethyl.

[0032] The term "C 3-7 "Cycloalkyl" means a monovalent saturated monocyclic or bicyclic hydrocarbon radical of 3 to 7 ring carbon atoms. Bicyclic means consisting of two saturated carbocyclic rings having one or two common carbon atoms. Examples of monocyclic cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. Examples of bicyclic cycloalkyl are bicyclo[1.1.0]butyl, bicyclo[2.2.1]heptyl, bicyclo[1.1.1]pentanyl or bicyclo[2.2.2]octyl.

[0033] The term "thiazolylene" refers to a divalent thiazolyl group.

[0034] The term "oxo" refers to a divalent oxygen atom =0.

[0035] The term "oxoimidazolidinyl" means

[0036] The term "haloazetidinyl" refers to an azetidinyl group in which at least one of the hydrogen atoms of the azetidinyl group has been replaced by the same or different halogen atoms (particularly, a fluorine atom). Examples of halogenated azetidinyl groups include fluoroazetidinyl and difluoroazetidinyl.

[0037] The term "2,2-propylene" means

[0038] The term "blocking group" means a group that selectively blocks a reactive site in a multifunctional compound so that a chemical reaction occurs selectively on another unprotected reactive site that is usually associated therewith in synthetic chemistry. The blocking group can be removed at an appropriate time point. Exemplary blocking groups are amino-protecting groups, carboxyl-protecting groups or hydroxyl-protecting groups.

[0039] Those skilled in the art will appreciate that the following structures of compounds of formula (Ia) and (Ia') are identical, especially with respect to the chiral centers:

[0040]

[0041] The term "pharmaceutically acceptable salts" refers to salts that are not biologically or otherwise undesirable. "Pharmaceutically acceptable salts" include both acid addition salts and base addition salts.

[0042] "Pharmaceutically acceptable acid addition salts" refer to those pharmaceutically acceptable salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and the like, and organic acids, which may be selected from aliphatic, alicyclic, aromatic, aromatic aliphatic, heterocyclic, formic and sulfonic acid organic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid, and the like.

[0043] The term "pharmaceutical base addition salt" refers to those pharmaceutical salts formed with organic or inorganic bases. Examples of acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese and aluminum salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include primary amines, secondary amines, tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines and basic ion exchange resins (such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, tromethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine and polyamine resins).

[0044] The term "pharmaceutically active metabolite" refers to a pharmacologically active product produced by the metabolism of a specific compound or its salt in the body. After entering the human body, most drugs are substrates for chemical reactions that may change their physical properties and biological effects. These metabolic transformations, which usually affect the polarity of the compounds of the present invention, change the way the drug is distributed in the body and excreted from the body. However, in some cases, drug metabolism is necessary for therapeutic effects.

[0045] The term "therapeutically effective amount" refers to an amount of a compound or molecule of the invention that, when administered to a subject, (i) treats or prevents a particular disease, disorder or condition, (ii) reduces, ameliorates or eliminates one or more symptoms of a particular disease, disorder or condition, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, disorder or condition as described herein. The therapeutically effective amount depends on the compound, the disease state being treated, the severity of the disease being treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending medical or veterinary doctor and other factors.

[0046] The term "pharmaceutical composition" means a mixture or solution comprising a therapeutically effective amount of an active drug ingredient together with a pharmaceutically acceptable excipient for administration to a mammal (eg, a human) in need thereof.

[0047] The terms "pharmaceutically acceptable excipient," "pharmaceutically acceptable carrier," and "therapeutically inert excipient" are used interchangeably and refer to any pharmaceutical ingredient in a pharmaceutical composition that is not therapeutically active and is non-toxic to the subject to which it is administered, such as a disintegrant, binder, filler, solvent, buffer, tonicity agent, stabilizer, antioxidant, surfactant, carrier, diluent, or lubricant used to formulate a drug product.

[0048] KRAS inhibitors

[0049] The present invention relates to (i) a compound of formula (I),

[0050]

[0051] in

[0052] R 1 for 3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazinyl or (C 1-6 Alkyl) oxoimidazolidinyl;

[0053] Where R 8 C 1-6 alkyl;

[0054] R 9 for ((C 1-6 alkyl)2amino)azetidinyl, C1-6 Alkylpiperazinyl, halogenated azetidinyl, halogenated C 1-6 Alkylamino, halogenated C 1-6 Alkylaminoazetidinyl, halo C 1-6 Alkylpiperazine, hydroxyl (C 1-6 alkyl)

[0055] piperidinyl or morpholinyl;

[0056] R 2 C 1-6 alkyl;

[0057] R 3 is H or halogen;

[0058] R 4 is H or halogen;

[0059] R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0060] R 6 C 1-6 Alkoxy C 1-6 alkyl;

[0061] R 7 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl;

[0062] A 1 is thiazolylidene;

[0063] A 2 C 1-6 Alkylene;

[0064] The prerequisite is R 3 and R 4 Not at the same time H;

[0065] or a pharmaceutically acceptable salt thereof.

[0066] Another embodiment of the present invention is (ii) a compound of formula (Ia),

[0067]

[0068] in

[0069] R 1 for 3-Oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazinyl

[0070] or (C 1-6Alkyl) oxoimidazolidinyl;

[0071] Where R 8 C 1-6 alkyl;

[0072] R 9 for ((C 1-6 alkyl)2amino)azetidinyl, C 1-6 Alkylpiperazinyl, halogenated azetidinyl, halogenated C 1-6 Alkylamino, halogenated C 1-6 Alkylaminoazetidinyl, halo C 1-6 Alkylpiperazine, hydroxyl (C 1-6 alkyl)piperidinyl or morpholinyl;

[0073] R 2 C 1-6 alkyl;

[0074] R 3 is H or halogen;

[0075] R 4 is H or halogen;

[0076] R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0077] R 6 C 1-6 Alkoxy C 1-6 alkyl;

[0078] R 7 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl;

[0079] A 1 is thiazolylidene;

[0080] A 2 C 1-6 Alkylene;

[0081] The prerequisite is R 3 and R 4 Not at the same time H;

[0082] or a pharmaceutically acceptable salt thereof.

[0083] A further embodiment of the present invention is (iii) a compound of formula (I) or (Ia) according to (i) or (ii), or a pharmaceutically acceptable salt thereof, wherein R 1 for Where R 8 C 1-6 Alkyl; R9 C 1-6 Alkylpiperazinyl, halogenated C 1-6 Alkylpiperazinyl or morpholinyl.

[0084] A further embodiment of the present invention is (iv) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (iii), wherein R 1 for Where R 8 is methyl; R 9 It is 4-methylpiperazin-1-yl, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or morpholinyl.

[0085] A further embodiment of the present invention is (v) a compound of formula (I) or (Ia) according to any one of (i) to (iv), wherein R 1 It is methyl-(4-methylpiperazine-1-carbonyl)amino, methyl-[4-(2,2,2-trifluoroethyl)piperazine-1-carbonyl]amino or methyl(morpholine-4-carbonyl)amino.

[0086] A further embodiment of the present invention is (vi) a compound of formula (I) or (Ia) according to any one of (i) to (v), or a pharmaceutically acceptable salt thereof, wherein R 2 It is isopropyl.

[0087] A further embodiment of the present invention is (vii) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (vi), wherein R 3 is H or fluorine.

[0088] A further embodiment of the present invention is (viii') a compound of formula (I) or (Ia) according to any one of (i) to (vii), or a pharmaceutically acceptable salt thereof, wherein R 3 For fluorine.

[0089] A further embodiment of the present invention is (ix) a compound of formula (I) or (Ia) according to any one of (i) to (viii), or a pharmaceutically acceptable salt thereof, wherein R 4 is H or fluorine.

[0090] A further embodiment of the present invention is (x) a compound of formula (I) or (Ia) according to any one of (i) to (ix), or a pharmaceutically acceptable salt thereof, wherein R 4 For H.

[0091] A further embodiment of the present invention is (xi) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (x), wherein R 5 Halogenated C 1-6alkyl.

[0092] A further embodiment of the present invention is (xii) a compound of formula (I) or (Ia) according to any one of (i) to (xi), or a pharmaceutically acceptable salt thereof, wherein R 5 It is 2,2,2-trifluoroethyl.

[0093] A further embodiment of the present invention is (xiii) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xii), wherein R 6 It is 1-methoxyethyl.

[0094] A further embodiment of the present invention is (xiv) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xiii), wherein R 7 For (halogenated C 1-6 alkyl)piperazinyl.

[0095] A further embodiment of the present invention is (xv) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xiv), wherein R 7 It is 4-(2,2,2-trifluoroethyl)piperazin-1-yl.

[0096] A further embodiment of the present invention is (xvi) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xv), wherein A 1 for Where bond "a" is attached to the indole ring.

[0097] A further embodiment of the present invention is (xvii) a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xvi), wherein A 2 It is 2,2-propylene.

[0098] Another embodiment of the present invention is (xviii) a compound of formula (I) or (Ia) according to (i) or (ii), wherein

[0099] R 1 for Where R 8 C 1-6 Alkyl; R 9 C 1-6 Alkylpiperazinyl, halogenated C 1-6 Alkylpiperazinyl or morpholinyl;

[0100] R 2 C 1-6 alkyl;

[0101] R 3is a halogen;

[0102] R 4 is H;

[0103] R 5 Halogenated C 1-6 alkyl;

[0104] R 6 C 1-6 Alkoxy C 1-6 alkyl;

[0105] R 7 For (halogenated C 1-6 alkyl)piperazinyl;

[0106] A 1 for wherein bond "a" is attached to the indole ring;

[0107] A 2 C 1-6 Alkylene;

[0108] or a pharmaceutically acceptable salt thereof.

[0109] Another embodiment of the present invention is (xix) a compound of formula (I) or (Ia) according to (xviii), wherein

[0110] R 1 is methyl-(4-methylpiperazine-1-carbonyl)amino, methyl-[4-(2,2,2-trifluoroethyl)piperazine-1-carbonyl]amino or methyl(morpholine-4-carbonyl)amino;

[0111] R 2 is isopropyl;

[0112] R 3 For fluorine;

[0113] R 4 is H;

[0114] R 5 It is 2,2,2-trifluoroethyl;

[0115] R 6 is (1S)-1-methoxyethyl;

[0116] R 7 is 4-(2,2,2-trifluoroethyl)piperazin-1-yl;

[0117] A 1 for wherein bond "a" is attached to the indole ring;

[0118] A 2 is 2,2-propylene;

[0119] or a pharmaceutically acceptable salt thereof.

[0120] The present invention relates to (i'), which is a compound of formula (I),

[0121]

[0122] in

[0123] R 1 for or (C 1-6 Alkyl) oxoimidazolidinyl;

[0124] Where R 8 C 1-6 alkyl;

[0125] R 9 for ((C 1-6 alkyl)2amino)azetidinyl, haloazetidinyl or morpholinyl;

[0126] R 2 C 1-6 alkyl;

[0127] R 3 is H or halogen;

[0128] R 4 is H or halogen;

[0129] R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0130] R 6 C 1-6 Alkoxy C 1-6 alkyl;

[0131] R 7 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl;

[0132] A 1 is thiazolylidene;

[0133] A 2 C 1-6 Alkylene;

[0134] The prerequisite is R 3 and R 4 Not at the same time H;

[0135] or a pharmaceutically acceptable salt thereof.

[0136] Another embodiment of the present invention is (ii') a compound of formula (Ia),

[0137]

[0138] in

[0139] R 1 for or (C 1-6 Alkyl) oxoimidazolidinyl;

[0140] Where R 8 C 1-6 alkyl;

[0141] R 9 for ((C 1-6 alkyl)2amino)azetidinyl, haloazetidinyl or morpholinyl;

[0142] R 2 C 1-6 alkyl;

[0143] R 3 is H or halogen;

[0144] R 4 is H or halogen;

[0145] R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0146] R 6 C 1-6 Alkoxy C 1-6 alkyl;

[0147] R 7 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl;

[0148] A 1 is thiazolylidene;

[0149] A 2 C 1-6 Alkylene;

[0150] The prerequisite is R 3 and R 4 Not at the same time H;

[0151] or a pharmaceutically acceptable salt thereof.

[0152] A further embodiment of the present invention is (iii') a compound of formula (I) or (Ia) according to (i') or (ii'), or a pharmaceutically acceptable salt thereof, wherein R1 for Where R 8 C 1-6 Alkyl; R 9 It is a halogenated azetidinyl or morpholinyl group.

[0153] A further embodiment of the present invention is (iv') a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i') to (iii'), wherein R 1 for Where R 8 is methyl; R 9 It is 3,3-difluoroazetidin-1-yl or morpholinyl.

[0154] A further embodiment of the present invention is (v') a compound of formula (I) or (Ia) according to any one of (i') to (iv'), wherein R 1 It is (3,3-difluoroazetidine-1-carbonyl)-methyl-amino or methyl(morpholine-4-carbonyl)amino.

[0155] A further embodiment of the present invention is (vi') a compound of formula (I) or (Ia) according to any one of (i') to (v'), or a pharmaceutically acceptable salt thereof, wherein R 2 It is isopropyl.

[0156] A further embodiment of the present invention is (vii') a compound of formula (I) or (Ia) according to any one of (i') to (vi'), or a pharmaceutically acceptable salt thereof, wherein R 3 is H or fluorine.

[0157] A further embodiment of the present invention is (viii') a compound of formula (I) or (Ia) according to any one of (i') to (vii'), or a pharmaceutically acceptable salt thereof, wherein R 3 For fluorine.

[0158] A further embodiment of the present invention is (ix') a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, according to any one of (i') to (viii'), wherein R 4 is H or fluorine.

[0159] A further embodiment of the present invention is (x') a compound of formula (I) or (Ia) according to any one of (i') to (ix'), or a pharmaceutically acceptable salt thereof, wherein R 4 For H.

[0160] A further embodiment of the present invention is (xi') a compound of formula (I) or (Ia) according to any one of (i') to (x'), or a pharmaceutically acceptable salt thereof, wherein R 5It is ethyl or 2,2,2-trifluoroethyl.

[0161] A further embodiment of the present invention is (xii') a compound of formula (I) or (Ia) according to any one of (i') to (xi'), or a pharmaceutically acceptable salt thereof, wherein R 6 It is 1-methoxyethyl.

[0162] A further embodiment of the present invention is (xiii') a compound of formula (I) or (Ia) according to any one of (i') to (xii'), or a pharmaceutically acceptable salt thereof, wherein R 7 It is 4-(2,2,2-trifluoroethyl)piperazin-1-yl or morpholinyl.

[0163] A further embodiment of the present invention is (xiv') a compound of formula (I) or (Ia) according to any one of (i') to (xiii'), or a pharmaceutically acceptable salt thereof, wherein A 1 for Where bond "a" is attached to the indole ring.

[0164] A further embodiment of the present invention is (xv') a compound of formula (I) or (Ia) according to any one of (i') to (xiv'), or a pharmaceutically acceptable salt thereof, wherein A 2 It is 2,2-propylene.

[0165] Another embodiment of the present invention is (xvi') a compound of formula (I) or (Ia) according to (i') or (ii'), wherein

[0166] R 1 for Where R 8 C 1-6 Alkyl; R 9 is a halogenated azetidinyl or morpholinyl group;

[0167] R 2 C 1-6 alkyl;

[0168] R 3 is a halogen;

[0169] R 4 is H;

[0170] R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0171] R 6 C 1-6 Alkoxy C 1-6 alkyl;

[0172] R7 is morpholinyl or (halogenated C 1-6 alkyl)piperazinyl;

[0173] A 1 for wherein bond "a" is attached to the indole ring;

[0174] A 2 C 1-6 Alkylene;

[0175] or a pharmaceutically acceptable salt thereof.

[0176] Another embodiment of the present invention is (xvii') a compound of formula (I) or (Ia) according to (xvi'), wherein

[0177] R 1 is (3,3-difluoroazetidine-1-carbonyl)-methyl-amino or methyl(morpholine-4-carbonyl)amino;

[0178] R 2 is isopropyl;

[0179] R 3 For fluorine;

[0180] R 4 is H;

[0181] R 5 is ethyl or 2,2,2-trifluoroethyl;

[0182] R 6 is (1S)-1-methoxyethyl;

[0183] R 7 is 4-(2,2,2-trifluoroethyl)piperazin-1-yl or morpholinyl;

[0184] A 1 for wherein bond "a" is attached to the indole ring;

[0185] A 2 is 2,2-propylene;

[0186] or a pharmaceutically acceptable salt thereof.

[0187] Another embodiment of the present invention is that (xx) is selected from the following compounds of formula (I) or (Ia):

[0188] 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide;

[0189] 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide;

[0190] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide;

[0191] 3-(Dimethylamino)-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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide;

[0192] 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide;

[0193] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide;

[0194] 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-tetraazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-3,3-difluoro-N-methyl-azetidine-1-carboxamide;

[0195] (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butanamide;

[0196] 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-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide;

[0197] N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide;

[0198] N-[(1S)-1-[[(7S,13S)-24-fluoro-20-(20M)-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide;

[0199] 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide;

[0200] (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-[methyl(2,2,2-trifluoroethylcarbamoyl)amino]butanamide;

[0201] 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-tetraazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(trifluoromethyl)azetidine-1-carboxamide;

[0202] (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazin-2-yl)butanamide;

[0203] (3R)-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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-3-hydroxy-N,3-dimethyl-piperidine-1-carboxamide;

[0204] N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide;

[0205] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N,4-dimethyl-piperazine-1-carboxamide; and

[0206] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(2,2,2-trifluoroethyl)piperazine-1-carboxamide;

[0207] or a pharmaceutically acceptable salt thereof.

[0208] Another embodiment of the present invention relates to (xxi) a method for preparing a compound according to any one of (i) to (xix) or (i') to (xvii'), comprising any of the following steps:

[0209] a) in the presence of a coupling agent and a base, a compound of formula (II),

[0210] With acid (III), A coupling reaction is carried out between them to form the compound of formula (I);

[0211] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , A 1 and A2 As defined in any one of (i) to (xix); the coupling reagent is T3P, HATU, PyBOP or EDCI / HOBt; and the base is TEA, DIEPA or DMAP.

[0212] Another embodiment of the present invention is (xxii) a compound or a pharmaceutically acceptable salt according to any one of (i) to (xx) or (i') to (xvii') for use as a therapeutically active substance.

[0213] Another embodiment of the present invention is (xxiii) a pharmaceutical composition comprising the compound according to any one of (i) to (xx) or (i') to (xvii') and a pharmaceutically acceptable excipient.

[0214] Another embodiment of the present invention is (xxiv) use of the compound according to any one of (i) to (xx) or (i') to (xvii') for treating a KRAS G12C protein-related disease.

[0215] Another embodiment of the present invention is (xxv) use of a compound according to any one of (i) to (xx) or (i') to (xvii') for treating KRAS G12C, G12D and G12V protein related diseases.

[0216] Another embodiment of the present invention is (xxvi) use of a compound according to any one of (i) to (xx) or (i') to (xvii') for inhibiting the interaction between RAS and downstream effectors, wherein the downstream effectors are RAF and PI3K.

[0217] Another embodiment of the present invention is (xxvii) use of a compound according to any one of (i) to (xx) or (i') to (xvii') for inhibiting propagation of oncogenic MAPK and PI3K signaling.

[0218] Another embodiment of the present invention is (xxviii) use of a compound according to any one of (i) to (xx) or (i') to (xvii') for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma, ovarian cancer and endometrial cancer.

[0219] Another embodiment of the present invention is (xxix) use of a compound according to any one of (i) to (xx) or (i') to (xvii') for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0220] Another embodiment of the present invention is (xxx) a compound or a pharmaceutically acceptable salt according to any one of (i) to (xx) or (i') to (xvii'), for use in treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0221] Another embodiment of the present invention is (xxxi) a compound or pharmaceutical salt as described in any one of (i) to (xx) or (i') to (xvii'), for use in treating or preventing a KRAS mutation-driven cancer, wherein the cancer comprises a first mutation of 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.

[0222] Another embodiment of the present invention is (xxxii) use of a compound according to any one of (i) to (xx) or (i') to (xvii') for the preparation of a medicament for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

[0223] Another embodiment of the present invention is (xxxiii) the use of a compound as described in any one of (i) to (xx) or (i') to (xvii') for the preparation of a medicament for treating or preventing a KRAS mutation-driven cancer, wherein the cancer comprises a first mutation of 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.

[0224] Another embodiment of the present invention is (xxxiv) a method for treating or preventing a KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer, the method comprising administering a therapeutically effective amount of a compound as defined in any one of (i) to (xx) or (i') to (xvii').

[0225] Another embodiment of the present invention is (xxxv) a method of treating or preventing a KRAS mutation-driven cancer, wherein the cancer comprises a first mutation of 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.

[0226] Another embodiment of the present invention is (xxxvi) a compound or a pharmaceutically acceptable salt according to any one of (i) to (xx) or (i') to (xvii'), which is prepared according to the method described in (xxi).

[0227] Pharmaceutical compositions and administration

[0228] Another embodiment provides a pharmaceutical composition or medicine containing the compounds of this invention and a therapeutic inert carrier, diluent or excipient, and methods for preparing such compositions and medicines using the compounds of this invention. In one example, the compound of formula (I) can be formulated as a galenical administration form by mixing with a physiologically acceptable carrier (i.e., a carrier that is nontoxic to the recipient at the dosage and concentration used) at ambient temperature under appropriate pH and desired purity. The pH of the preparation depends primarily on the specific use and concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, the compound of formula (I) is prepared in an acetate buffer at pH 5. In another embodiment, the compound of formula (I) is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized preparation, or as an aqueous solution.

[0229] The composition is formulated, metered, and administered in a manner consistent with good medical practice. Factors to be considered in this case include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the delivery site of the agent, the method of administration, the timing of administration, and other factors known to practitioners. The "effective amount" of the compound to be administered will be affected by such considerations and is the minimum amount necessary to inhibit the interaction of mutant RAS (e.g., KRAS G12C) with RAF, thereby blocking oncogenic MAPK signaling. For example, the amount may be lower than an amount that is toxic to normal cells or mammals as a whole.

[0230] In one example, the pharmaceutically effective amount of the compound of the present invention administered parenterally per dose will be in the range of about 0.1 to 1000 mg / kg patient body weight per day, alternatively about 0.1 to 1000 mg / kg patient body weight, typically an initial range of 0.3 to 15 mg / kg / day of the compound used. In another embodiment, oral unit dosage forms such as tablets and capsules preferably contain about 1 to about 1000 mg of the compound of the present invention.

[0231] The compounds of the present invention can be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and (if necessary for local treatment) intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

[0232] The compounds of the present invention can be administered in any convenient administration form, for example, tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain conventional components of pharmaceutical preparations, for example, diluents, carriers, pH adjusters, sweeteners, fillers and other active agents.

[0233] Typical formulations are prepared by mixing the compounds of the invention with carriers or excipients. Suitable carriers and excipients are well known to those skilled in the art and are described in, for example, Ansel, Howard C. et al., Ansel' sPharmaceuticalDosage Forms and DrugDeliverySystems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R. et al. Remington: The Science and Practice of Pharmacy .Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients .Chicago, Pharmaceutical Press, 2005. The formulation may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavoring agents, flavoring agents, diluents and other known additives to provide an aesthetically pleasing display of a drug (e.g., a compound of the present invention or a pharmaceutical composition thereof) or to facilitate the preparation of a pharmaceutical product (e.g., a drug).

[0234] An example of a suitable oral dosage form is a tablet containing about 1 to 1000 mg of a compound of the present invention and about 1 to 1000 mg of anhydrous lactose, about 1 to 1000 mg of cross-linked sodium carboxymethylcellulose, about 1 to 1000 mg of polyvinyl pyrrolidone (PVP) K30 and about 1 to 1000 mg of magnesium stearate. The powdered ingredients are first mixed together and then mixed with the PVP solution. The resulting composition can be dried, granulated, mixed with magnesium stearate and compressed into tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound of the present invention (e.g., 5 mg to 400 mg) in a suitable buffer solution (e.g., phosphate buffer) and adding a penetration enhancer (e.g., a salt such as sodium chloride) if necessary. The solution can be filtered, for example, using a 0.2 micron filter to remove impurities and contaminants.

[0235] Thus, embodiments include a pharmaceutical composition comprising a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof. Further embodiments include a pharmaceutical composition comprising a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0236] Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in treating a mutant KRAS driven cancer. Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in treating a mutant KRAS driven cancer.

[0237] The following compositions A and B illustrate typical compositions of the present invention but are intended to be representative thereof only.

[0238] Composition A

[0239] The compounds of the invention can be used as active ingredients in a manner known per se to produce tablets of the following composition:

[0240]

[0241] Composition B

[0242] The compounds of the invention can be used as active ingredients in a manner known per se to produce capsules of the following composition:

[0243]

[0244] Indications and treatments

[0245] The compounds of the present invention induce new binding pockets in KRAS by driving the formation of a high-affinity tri-complex between KRAS protein and widely expressed cyclophilin A (CYPA), and these compounds inhibit the interaction of KRAS with downstream effectors (such as RAF and PI3K). Therefore, the compounds of the present invention can be used to inhibit the propagation of oncogenic MAPK and PI3K signal transduction, reducing cell (especially cancer cell) proliferation. The compounds of the present invention can be used to terminate RAS signal transduction in cells of pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma, ovarian cancer, endometrial cancer, etc. driven by RAS mutants (especially KRAS mutations). Alternatively, the compounds of the present invention can be used to terminate RAS signal transduction in malignant solid tumors, wherein the carcinogenic effect of KRAS mutations is enhanced by the deregulation or mutation of effector pathways such as signal transduction driven by MAPK, PI3K-AKT-mTOR (mammalian rapamycin target), and these compounds are used for targeted therapies of pancreatic adenocarcinoma, colorectal cancer, non-small cell lung cancer, etc.

[0246] Another embodiment includes a method of treating or preventing cancer in a mammal in need of such treatment, wherein the method comprises administering to said mammal a therapeutically effective amount of a compound of formula (I), a stereoisomer, tautomer, prodrug or pharmaceutically acceptable salt thereof.

[0247] synthesis

[0248] The compounds of the present invention may be prepared by any conventional method. Suitable methods for synthesizing these compounds and their starting materials are provided in the following schemes and examples. Unless otherwise indicated, all substituents, especially R 1 To R 7 and A 1 and A 2 As defined above. Furthermore, unless otherwise specifically indicated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to one of ordinary skill in the art of organic chemistry.

[0249] The general synthetic route for preparing compounds of formula (I) is shown below.

[0250] Solution 1

[0251]

[0252] The compound of formula II is synthesized according to the procedures described in Intermediates A to J. The compound of formula (I) can be obtained by coupling reaction between acid (III) and compound of formula (II) using one or more coupling reagents (such as T3P, HATU, PyBOP and EDCI / HOBt) in the presence of a base (such as TEA, DIEPA and DMAP).

[0253] The compounds of the present invention can be obtained as diastereomers or mixtures of enantiomers, which can be separated by methods well known in the art, for example, (chiral) HPLC or SFC. In another embodiment, compounds of formula (I) can be obtained according to the above scheme using the corresponding chiral starting materials.

[0254] The present invention also relates to a method for preparing a compound of formula (I), the method comprising the following steps:

[0255] a) in the presence of a coupling agent and a base, a compound of formula (II),

[0256] With acid (III),

[0257] A coupling reaction is carried out between them to form the compound of formula (I);

[0258] in

[0259] In step a), the coupling reagent may be, for example, T3P, HATU, PyBOP or EDCI / HOBt; the base may be, for example, TEA, DIEPA or DMAP.

[0260] The compounds of formula (I) or (Ia) produced according to the above-described process are also an object of the present invention.

[0261] Examples

[0262] The present invention will be more fully understood by reference to the following examples. However, they should not be construed as limiting the scope of the present invention.

[0263] abbreviation

[0264] The present invention will be more fully understood by reference to the following examples. However, they should not be construed as limiting the scope of the present invention.

[0265] The abbreviations used in this article are as follows:

[0266] ACN Acetonitrile

[0267] aq. water-based

[0268] BOC-L-Valine (S)-2-((tert-Butyloxycarbonyl)amino)-3-methylbutanoic acid

[0269] Boc-N-Me-Val-OH N-(tert-Butyloxycarbonyl)-N-methyl-L-valine

[0270] (Boc)2O Di-tert-butyl dicarbonate (R)-Diphenylphosphine (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl

[0272] CDCl3: deuterated chloroform

[0273] CDI 1,1'-Carbonyldiimidazole

[0274] CD3OD: deuterated methanol

[0275] COMU (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbonium hexafluorophosphate

[0276] DIEPA: N,N-diethylpropylamine

[0277] DIBAL-H Diisobutylaluminum hydride

[0278] DMAP: 4-dimethylaminopyridine

[0279] DMF: Dimethylformamide

[0280] DMP 1,1,1-tri(acetoxy)-1,1-dihydro-1,2-benzimidoyl-3-(1H)-one

[0281] DMSO: dimethyl sulfoxide

[0282] EDCI: N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride

[0283] EtOAc or EA: Ethyl acetate

[0284] FRET Fluorescence Resonance Energy Transfer

[0285] HATU: (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate)

[0286] hr(s): hours

[0287] HPLC: High Performance Liquid Chromatography

[0288] HOBt: N-Hydroxybenzotriazole

[0289] H-VAL-OTBU HCl (S)-tert-Butyl-2-amino-3-methylbutyric acid methyl ester hydrochloride [Ir(OMe)(COD)]2 (1,5-cyclooctadiene)(methoxy)iridium(I) dimer LDA Lithium diisopropylamide

[0290] MS: (ESI): Mass spectrometry (electrospray ionization)

[0291] min(s) minutes

[0292] MTBE Methyl tert-butyl ether

[0293] NMM N-Methylmorpholine

[0294] NaBH(OAc)3 Sodium triacetoxyborohydride NMR: Nuclear Magnetic Resonance

[0295] NMO 4-Methylmorpholine N-oxide obsd. Observed value

[0296] Pd(dppf)Cl2 [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II) chloride

[0297] Pd(dtbpf)Cl2 [1,1'-Bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II)

[0298] prep-HPLC Preparative High Performance Liquid Chromatography PyBOP: Benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate

[0299] RT or rt: room temperature

[0300] sat. saturated

[0301] SFC Supercritical Fluid Chromatography

[0302] TEA: Triethylamine

[0303] TFA: trifluoroacetic acid

[0304] THF: Tetrahydrofuran

[0305] TEA: Trimethylamine

[0306] TMEDA Tetramethylethylenediamine

[0307] TMSCF3 Trifluoromethyltrimethylsilane T3P: Propylphosphonic anhydride

[0308] General experimental conditions

[0309] Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and Quad 12 / 25 Cartridge module. ii) ISCO combi-flash chromatograph. Silica gel brand and pore size: i) KP-SIL Particle size: 40-60 μm; ii) CAS registration number: silica gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Ocean Chemical Co., Ltd., pore size: 200-300 or 300-400.

[0310] Intermediates and final compounds were purified by preparative HPLC on a reverse phase column using XBridge TM Prep-C18 (5μm, OBDTM 30×100mm) column, SunFire TM Prep-C18(5μm,OBD TM30×100mm) chromatographic column, Phenomenex Synergi-C18 (10μm, 25×150mm) or Phenomenex Gemini-C18 (10μm, 25×150mm). Waters AutoP purification system (sample manager 2767, pump 2525, detector: Micromass ZQ and UV 2487, solvent system: acetonitrile and 0.1% ammonium hydroxide in water; acetonitrile and 0.1% FA in water, or acetonitrile and 0.1% TFA in water). Or Gilson-281 purification system (pump 322, detector: UV 156, solvent system: acetonitrile and 0.05% ammonium hydroxide in water; acetonitrile and 0.225% FA in water; acetonitrile and 0.05% HCl in water; acetonitrile and 0.075% TFA in water; or acetonitrile and water).

[0311] For SFC chiral separation, the intermediates were separated by chiral columns (Daicel chiralpak IC, 5 μm, 30×250 mm), AS (10 μm, 30×250 mm) or AD (10 μm, 30×250 mm), using Mettler Toledo Multigram III system SFC, Waters 80Q preparative SFC or Thar 80 preparative SFC, solvent system: CO2 and IPA (0.5% TEA in IPA solution) or CO2 and MeOH (0.1% NH3·H2O in MeOH solution), back pressure 100 bar, UV detection at 254 or 220 nm.

[0312] Using LC / MS (Waters TM The LC / MS spectra of the compounds were obtained using an Agilent Alliance 2795-Micromass ZQ, Shimadzu Alliance2020-Micromass ZQ or Agilent Alliance 6110-Micromass ZQ. The LC / MS conditions were as follows (run time 3 or 1.5 min):

[0313] Acidic conditions I: A: 0.1% TFA solution in H2O; B: 0.1% TFA solution in acetonitrile;

[0314] Acidic conditions II: A: 0.0375% TFA solution in H2O; B: 0.01875% TFA solution in acetonitrile;

[0315] Basic conditions I: A: 0.1% NH3·H2O solution in H2O; B: acetonitrile;

[0316] Basic conditions II: A: 0.025% NH3·H2O solution in H2O; B: acetonitrile;

[0317] Neutral conditions: A: H2O; B: acetonitrile.

[0318] Mass spectra (MS): Usually only ions representing the parent mass are reported, and unless otherwise stated, the mass ion quoted is the positive mass ion (MH) + .

[0319] NMR spectra were obtained using a Bruker Avance 400 MHz or 500 MHz.

[0320] Microwave-assisted reactions were performed in a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were performed under argon or nitrogen atmosphere. Unless otherwise stated, reagents were purchased from commercial suppliers as received without further purification.

[0321] Preparation Example

[0322] The following examples are intended to illustrate the meaning of the present invention, but are by no means intended to limit the meaning of the present invention: Preparation of intermediates

[0323] Intermediate A

[0324] 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-methyl-piperidin

[0325]

[0326] The title intermediate A was prepared according to the following scheme:

[0327]

[0328]

[0329] 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)

[0330] To a solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (Compound A1, 2.0 g, 9.26 mmol) and bis(pinacol)diboron (3.5 g, 13.9 mmol) in THF (30 mL) was 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 under N2 for 16 hours. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (EA / PE: 0-20%) to give 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (Compound A2, 2.4 g) as a yellow oil. 1 HNMR (400MHz, CDCl3) δppm 8.91 (d, J = 1.4Hz, 1H), 8.21 (d, J = 1.4Hz, 1H), 4.95 (q, J = 6.5Hz, 1H), 3.30 (s, 3H), 1.49 (d, J = 6.5Hz, 3H), 1.35 (s, 12H).

[0331] Step 2: Preparation of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3)

[0332] 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 hours under N2 protection. The reaction was quenched with saturated Na2SO3 solution (40 mL) and the reaction mixture was extracted with EtOAc (30 mL, twice). The combined organic layers were washed with brine (50 mL), filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA / PE: 0-20%) to give 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3, 660 mg) as a yellow oil. MS calculated value 342 (MH + ); measured value 341.8 (MH + ).

[0333] Step 3: Preparation of benzyl 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylate (Compound A5)

[0334] To a solution of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3, 660 mg, 1.9 mmol) and 1-Cbz-piperazine (Compound A4, 425.1 mg, 1.9 mmol) in toluene (10 mL) was added cesium carbonate (1.6 g, 4.83 mmol), (R)-BINAP (60.1 mg, 0.1 mmol) and palladium (II) acetate (43.3 mg, 0.19 mmol). The mixture was stirred at 100 ° C. for 12 hours under N2. The mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA / PE: 0-50%) to give 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (Compound A5, 740 mg) as a yellow solid. MS calculated value 434.1 (MH + ); measured value 434.1 (MH + ).

[0335] Step 4: Preparation of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-methyl-piperazine (Intermediate A)

[0336] To a solution of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound A5, 740 mg, 1.7 mmol) and bis(pinacol)borate (519.2 mg, 2.04 mmol) in toluene (12 mL) was 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 under N2 protection for 12 hours. 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-pyridinyl]-4-methyl-piperazine (Intermediate A, 470 mg) as a brown solid. MS calculated value 482.3 (MH + ); measured value 482.2 (MH + ).

[0337] Intermediate B

[0338] (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid methyl ester

[0339]

[0340] Intermediate B was prepared according to the following scheme:

[0341]

[0342] Step 1: Preparation of (4-bromothiazol-2-yl)methanol (Compound B2)

[0343] 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, three times). The combined organic phases were washed twice with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (4-bromothiazol-2-yl)methanol (Compound B2, 6 g) as a colorless oil.

[0344] Step 2: Preparation of 4-bromo-2-(bromomethyl)thiazole (Compound B3)

[0345] To a solution of (4-bromothiazol-2-yl)methanol (compound B2, 6.0 g, 30.92 mmol) in DCM (80 mL) was added CBr4 (15.4 g, 46.38 mmol) and triphenylphosphine (12.1 g, 46.38 mmol) at 0°C. After stirring at 25°C for 1 hour, the mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column eluting with ethyl acetate (0-10%) in petroleum ether to give (4-bromothiazol-2-yl)methanol (compound B3, 6.0 g) as a yellow oil. MS calculated value 255.9 (MH + ); measured value 255.9 (MH + ).

[0346] Step 3: Preparation of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5)

[0347] To a mixture of (R)-2,5-dihydro-3,6-dimethoxy-2-isopropylpyrazine (Compound B4, 4.3 g, 23.45 mmol) in THF (60 mL) was slowly added n-butyllithium (10 mL, 25.22 mmol, 2.5 M) at -78 °C. After addition, the mixture was stirred at -78 °C for 0.5 hours. 4-Bromo-2-(bromomethyl)thiazole (Compound B3, 5.4 g, 21.02 mmol) was added to the above mixture at -78 °C, and the mixture was stirred for another hour. The reaction was quenched with saturated NH4Cl solution (100 mL) and the reaction mixture was extracted with EtOAc (100 mL, twice). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by reverse phase chromatography to give 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5, 3.6 g) as a yellow oil. MS calculated 360 (MH + ); measured value 359.9 (MH + ).

[0348] Step 4: Preparation of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoate (Compound B6)

[0349] To a solution of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (Compound B5, 3.6 g, 10 mmol) in ACN (20 mL) was added hydrochloric acid (66.6 mL, 0.3 M). The mixture was stirred at 25 ° C for 2 hours. The mixture was basified with saturated NaHCO3 solution until pH = 8. The mixture was extracted twice with EtOAc (80 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoic acid methyl ester (Compound B6, 3.1 g) as a yellow oil. MS calculated value 264.9 (MH + ); measured value 264.9 (MH + ).

[0350] Step 5: Preparation of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoate (Compound B7)

[0351] To a solution of (2S)-2-amino-3-(4-bromothiazol-2-yl)propanoic acid methyl ester (Compound B6, 3.1 g, 11.69 mmol) in DCM (40 mL) was added triethylamine (2.9 g, 29.23 mmol) and (Boc)2O (3.8 g, 17.54 mmol). After stirring at 30 °C for 12 hours, the mixture was concentrated under vacuum. The residue was purified by silica gel column eluted with ethyl acetate (0-30%) in petroleum ether to give (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid methyl ester (Compound B7, 3.2 g) as a yellow oil. MS calculated value 387 (MNa + ); measured value 386.9 (MNa + ).

[0352] Step 6: Preparation of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (Compound B8)

[0353] To a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid methyl ester (Compound B7, 3.2 g, 8.76 mmol) in THF (30 mL), methanol (2 mL) and water (10 mL) was added lithium hydroxide (0.4 mL, 43.81 mmol). After stirring at 25 ° C for 1 hour, the reaction mixture was acidified by 1M HCl solution until pH = 5. The mixture was extracted twice with EtOAc (40 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid (Compound B8, 3.1 g) as a yellow oil. MS calculated value 373 (MNa + ); measured value 372.9 (MNa + ).

[0354] Step 7: Preparation of methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (Intermediate B)

[0355] 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) was added (3S)-methyl 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 hour, the reaction mixture was diluted with water (60 mL) and extracted with EtOAc (60 mL, three times). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column and eluted with ethyl acetate (10-30%) in petroleum ether to give (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Intermediate E, 2.4 g). MS calculated value 477 (MH + ), measured value 476.9 (MH + ).

[0356] Intermediate C

[0357] (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0358]

[0359] The title intermediate C was prepared according to the following scheme:

[0360]

[0361]

[0362] Step 1: Preparation of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (Compound C3)

[0363] 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, a solution of SnCl4 (97.2 mL, 121.5 mmol) was slowly added. After the mixture was stirred at -40 ° C for 0.5 hours, 5-bromo-6-fluoro-1H-indole (compound C2, 25.0 g, 116.8 mmol) in DCM (200 mL) was added dropwise, and the mixture was stirred at -40 ° C for another 15 min. After the reaction was completed, it was quenched with saturated NaHCO3 aqueous solution (800 mL), and the reaction mixture was extracted with EtOAc (900 mL, twice). The combined organic layer was washed with brine (700 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was ground with a solution (100 mL, petroleum ether: ethyl acetate = 8: 1) and filtered. The filter cake was dried in vacuo to give 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (Compound C3, 50.0 g) as a yellow solid. MS calculated 552.1 (MH + ); measured value 552.1 (MH + ).

[0364] Step 2: Preparation of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C4)

[0365] 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 LiBH4 (48.4 mL, 193.49 mmol, 4 M in THF) dropwise at 0°C. The mixture was stirred at 70°C under a nitrogen atmosphere for 24 hours. After the reaction was complete, it was quenched by slowly adding water (600 mL) at 0°C, and the reaction mixture was extracted with EtOAc (600 mL, twice). The combined organic layers were washed with brine (600 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 20% to 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 + ).

[0366] 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)

[0367] Silver trifluoromethanesulfonate (20.3 g, 78.88 mmol) was added to a mixture of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound C4, 35.4 g, 65.73 mmol) and iodine (18.4 g, 72.3 mmol) in THF (400 mL) at 0 ° C. The mixture was stirred at 0 ° C for 10 minutes. After the reaction was completed, it was quenched by saturated Na2SO3 aqueous solution (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% to 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 664.0 (MH + ); measured value 664.1 (MH + ).

[0368] Step 4: Preparation of 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silanyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C6)

[0369] To a mixture of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 16.7 g, 25.13 mmol) and 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (Intermediate A, 16.7 g, 34.69 mmol) in a mixed solution of 1,4-dioxane (270 mL) / toluene (90 mL) / water (90 mL) were added potassium phosphate (15.7 g, 73.92 mmol) and Pd(dppf)Cl2 (920 mg, 1.26 mmol). The mixture was stirred at 70° C. for 12 hours under a nitrogen atmosphere. After the reaction was completed, the mixture was filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 20% to 50%) to give 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silanyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylate (Compound C6, 19.5 g) as a white solid. MS calculated value 891.3 (MH + ); measured value 891.3 (MH + ).

[0370] Step 5: Preparation of 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silanyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C7)

[0371] To a solution of 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound C6, 14.5 g, 16.26 mmol) and Cs2CO3 (15.9 g, 48.77 mmol) in DMF (200 mL) was added 2,2,2-trifluoroethyl trifluoromethanesulfonate (37.7 g, 162.56 mmol) dropwise at 0 ° C, and the mixture was stirred at 20 ° C for 12 hours. After the reaction was complete, EtOAc (70 mL) and water (100 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (70 mL, twice). The combined organic layers were washed with brine (100 mL, four times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica gel column chromatography to give 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silanyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (Compound C7, 8.0 g, faster elution) as a yellow oil. MS calculated value 973.3 (MH + ); measured value 973.2 (MH + ).

[0372] Step 6: Preparation of 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C8)

[0373] To a solution of 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound C7, 10.5 g, 10.78 mmol) in DMF (130 mL) was added cesium fluoride (8.2 g, 53.9 mmol), and the mixture was stirred at 60 ° C for 24 hours. After the reaction was complete, EtOAc (100 mL) and water (100 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (100 mL, twice). The combined organic layers were washed with brine (80 mL, three times), 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% to 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-pyridinyl]piperazine-1-carboxylate (Compound C8, 6.5 g) as a yellow solid. MS calculated value 735.2 (MH + ); measured value 735.1 (MH + ).

[0374] Step 7: Preparation of 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C9)

[0375] To a solution of 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (Compound C8, 5.4 g), bis(pinacol)borate (2.8 g, 11.01 mmol) and potassium acetate (1.2 mL, 18.35 mmol) in toluene (70 mL) was added Pd(dppf)Cl2 (537.1 mg, 0.73 mmol). The mixture was degassed and purged with a nitrogen atmosphere three times, and the mixture was stirred at 90 ° C for 12 hours. After the reaction was complete, the mixture was cooled to room temperature. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 25% to 66%) to give 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C9, 5.2 g) as a yellow oil. MS calculated value 783.3 (MH + ); measured value 783.3 (MH + ).

[0376] Step 8: Preparation of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound C10)

[0377] Under nitrogen atmosphere, (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Intermediate B, 2.7 g, 5.69 mmol), 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2 To a mixture of toluene (60 mL) / 1,4-dioxane (20 mL) / water (20 mL) was added K3PO4 (3.4 g, 15.81 mmol) and Pd(dtbpf)Cl2 (412.2 mg, 0.63 mmol). The mixture was stirred at 70 ° C for 12 hours. After the reaction was complete, the mixture was concentrated in vacuo to give a residue. The residue was purified by silica gel column (EtOAc in PE = 10% to 75%) to give (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound C10, 3.6 g) as a brown solid. MS calculated value 1053.4 (MH + ); measured value 1053.3 (MH + ).

[0378] 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)

[0379] To a solution of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound C10, 3.6 g, 3.42 mmol) in DCE (50 mL) was added trimethyltin hydroxide (2.4 g, 13.67 mmol), and the mixture was stirred at 60 ° C for 12 hours. After the reaction was complete, EtOAc (80 mL) and water (60 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (80 mL, twice). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under vacuum to give (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxylic acid (Compound C11, 4.3 g) as a brown solid. MS calculated value 1039.4 (MH + ); measured value 1039.2 (MH + ).

[0380] Step 10: Preparation of 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2 ,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C12)

[0381] To a mixture of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid (compound C11, 4.3 g, 4.14 mmol) in DCM (430 mL) was added DIEA (14.4 mL, 82.76 mmol), EDCI (11.9 g, 62.07 mmol) and 1-hydroxybenzotriazole (1.4 g, 10.35 mmol) at 0° C. The mixture was stirred at 15° C. for 12 hours. After the reaction was complete, the mixture was concentrated in vacuo, then diluted with water (80 mL) and extracted with EtOAc (80 mL, twice). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc in PE = 25% to 66%) to give 4-[5-[(7S, 13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridinyl]piperazine-1-carboxylic acid benzyl ester (Compound C12, 3.1 g). MS calculated value 1021.4 (MH + ); measured value 1021.2 (MH + ).

[0382] Step 11: Preparation of N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 tert-Butyl]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound C13)

[0383] To 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22 ,26 [] octadecanoic acid-1 (25), 2, 5 (28), 19, 22 (26), 23-hexaen-(20M)-20-yl] -6- [(1S) -1-methoxyethyl] -3-pyridyl] piperazine-1-carboxylic acid benzyl ester (compound C12, 3.1 g, 3.04 mmol) and formaldehyde solution (775.0 mg, 9.55 mmol) in methanol (150 mL) were added with activated carbon-supported Pd (OH) 2 (2.79 g, 3.97 mmol). The mixture was degassed and purged with H 2 three times. The mixture was hydrogenated at 30 ° C for 18 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated in vacuo to give N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a brown solid. 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamic acid tert-butyl ester (Compound C13, 2.6 g). MS calculated value 901.3 (MH + ); measured value 901.3 (MH + ).

[0384] Step 12: Preparation of (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C)

[0385] To N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 To a mixture of tert-butyl]octadecanoate-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (compound C13, 2.6 g, 2.89 mmol) in DCM (18 mL) was added TFA (14.0 mL, 181.72 mmol). The mixture was stirred at 15 ° C for 0.5 h. After the reaction was complete, the mixture was concentrated in vacuo and diluted with saturated NaHCO (30 mL) and extracted with EtOAc (30 mL, three times). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow solid. 2,5 .1 9,13 .0 22 ,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C, 2.0 g), which was used directly in the next step. MS calculated value 801.3 (MH + ); measured value 801.2 (MH + )

[0386] Intermediate D

[0387] (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0388]

[0389] The title compound was prepared analogously to the preparation of Intermediate C by using iodoethane instead of 2,2,2-trifluoroethyl triflate.

[0390] Intermediate E

[0391] (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0392]

[0393] The compounds were prepared according to the following scheme:

[0394]

[0395]

[0396] Step 1: Preparation of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2).

[0397] To a mixture of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (Compound A3, 2.03 g, 5.95 mmol) and 1-(2,2,2-trifluoroethyl)piperazine (Compound E1, 1.0 g, 5.95 mmol) in toluene (15 mL) was added CsCO (4.85 g, 14.88 mmol), (R)-binap (92.6 mg, 0.15 mmol) and Pd(OAc) (66.8 mg, 0.3 mmol). The reaction mixture was degassed and purged with nitrogen 3 times, and the mixture was stirred at 100 ° C for 12 hours under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography to give 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2, 2.0 g) as a yellow oil. MS calculated value 382.2 (MH +); measured value 382.1 (MH + ).

[0398] Step 2: 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3).

[0399] To a solution of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E2, 3.2 g, 8.37 mmol), bis(pinacol)borate (3.19 g, 12.56 mmol) and KOAc (2.1 g, 20.93 mmol) in toluene (50 mL) was added Pd(dppf)Cl2 (306.3 mg, 0.42 mmol). The mixture was degassed and purged with nitrogen three times, and the mixture was stirred at 90° C. under a nitrogen atmosphere for 12 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue, which was purified by reverse phase column to give 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3, 1.9 g) as a yellow gum. MS calculated value 430.2 (MH + ), measured value 348.4(M-C6H 10 +H + ).

[0400] Step 3: Preparation of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E4).

[0401] To a solution of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]-4-(2,2,2-trifluoroethyl)piperazine (Compound E3, 1.9 g, 4.41 mmol), [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5, 2.1 g, 3.15 mmol) in 1,4-dioxane (24 mL), water (8 mL) and toluene (8 mL) was added K3PO4 (2.1 g, 9.5 mmol) and Pd(dppf)Cl2 (231 mg, 0.37 mmol). The mixture was degassed by bubbling nitrogen for 2 min, and the reaction mixture was stirred at 70°C for 12 h. After cooling to room temperature, the reaction mixture was filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE: 30%-60%) to give [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-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 + ).

[0402] Step 4: Preparation of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E5).

[0403] To a solution of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E4, 1 g, 1.14 mmol) in DMF (35 mL) was added CsCO (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, three times). The combined organics were washed with brine (50 mL, three times), dried over Na2SO4, filtered, and concentrated in vacuo to give a residue that was purified by column chromatography (EtOAc in PE: 30%-40%) to give [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E5, 640.0 mg, 0.69 mmol, eluted faster) as a white solid. MS calcd. 921.3 (MH + ); measured value 921.4 (MH + ).

[0404] Step 5: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E6).

[0405] To a solution of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound E5, 640.0 mg, 0.69 mmol) in DMF (7 mL) was added cesium fluoride (421.8 mg, 2.78 mmol). The mixture was stirred at 60 ° C for 16 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE: 30%-60%) to give 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E6, 360.0 mg) as a yellow oil. MS calculated 683.2 (MH + ); measured value 683.1 (MH + ).

[0406] Step 6: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E7).

[0407] To a solution of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E6, 360.0 mg, 0.53 mmol), bis(pinacol)boronate (200.6 mg, 0.79 mmol) in toluene (6 mL) was added potassium acetate (129 mg, 1.32 mmol) and Pd(dppf)Cl2 (40 mg, 0.1 mmol). The reaction mixture was degassed by bubbling nitrogen for 5 min 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-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E7, 300.0 mg) as a yellow gum. MS calcd. 731.4 (MH + ); measured value 731.4 (MH + ).

[0408] Step 7: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound E8).

[0409] To 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound E7, 0.3 g, 0.41 mmol) and (3S)-1-[(2S)-3-(4-bromothiophene)-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethylpropan-1-ol (Compound E7, 0.3 g, 0.41 mmol) were added. To a mixture of toluene (3 mL), 1,4-dioxane (1 mL) and water (1 mL) was 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 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. The residue was purified by column chromatography (EtOAc in PE: 60%-80%) to give (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound E8, 200.0 mg) as a yellow gum. MS calcd. 1001.4 (MH + ); measured value 1001.4 (MH + ).

[0410] Step 8: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound E9).

[0411] To a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound E8, 200.0 mg, 0.2 mmol) in DCE (5 mL) was added Me3SnOH (200.0 mg, 1.11 mmol). The mixture was stirred at 60 ° C for 12 hours. The reaction mixture was concentrated under vacuum to give a residue. EtOAc (10 mL) and water (10 mL) were added to the residue, and the layers were separated. The aqueous phase was extracted with EtOAc (15 mL, twice). The combined organic layers were washed with brine (20 mL), 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-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound E9, 188.0 mg) as a brown solid. MS calculated value 987.4 (MH + ); measured value 987.4 (MH + ).

[0412] Step 9: Preparation of N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]tert-butyl octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound E10).

[0413] To a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (compound E9, 188.0 mg, 0.19 mmol) in DCM (20 mL) at 0°C were added DIEA (0.7 mL, 3.81 mmol), EDCI (550.0 mg, 2.87 mmol) and HOBt (65.0 mg, 0.48 mmol). After stirring at 20° C. for 12 h, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (20 mL, three times). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue, which was purified by column chromatography (EtOAc in PE: 50%-70%) to give N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow solid. 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamic acid tert-butyl ester (Compound E10, 110.0 mg). MS calculated value 969.4 (MH + ); measured value 969.5 (MH + ).

[0414] Step 10: Preparation of (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E).

[0415] To N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 To a solution of tert-butyl]octadecanoate-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (compound E10, 110.0 mg, 0.11 mmol) in DCM (1 mL) was added TFA (1.0 mL, 12.98 mmol). The mixture was stirred at 20 ° C for 1 h. After the reaction was complete, the reaction mixture was concentrated under vacuum to give a residue. Saturated NaHCO3 aqueous solution (20 mL) was added and the mixture was extracted with EtOAc (15 mL, three times). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow solid. 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E, 98.0 mg). MS calculated value 869.4 (MH + ); measured value 869.2 (MH + ).

[0416] Intermediate F

[0417] (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0418]

[0419] The title compound was prepared analogously to the preparation of intermediate E by using iodoethane instead of 2,2,2-trifluoroethyl triflate.

[0420] Intermediate G

[0421] (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 2,5 .1 9 ,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0422]

[0423]

[0424] Step 1: Preparation of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridinyl]morpholine (Compound G1)

[0425] 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) was 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 3 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 obtain a residue. The residue was purified by column chromatography to obtain 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 + ).

[0426] 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)

[0427] To a solution of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (Compound G1, 21 g, 63.3 mmol), bis(pinacol)borate (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 under a nitrogen atmosphere for 12 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give 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 + ).

[0428] Step 3: Preparation of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound G3)

[0429] To a solution of 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridinyl]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) and water (80 mL) was added K3PO4 (29.7 g, 2.33 mmol) and Pd(dppf)Cl2 (1.7 g, 0.29 mmol). The mixture was degassed by bubbling nitrogen for 2 min, and the reaction mixture was stirred at 90°C for 18 hours. After cooling to room temperature, the reaction mixture is extracted with EA (200mL, three times). The combined organic layer is washed with brine (200mL), dried over Na2SO4, filtered and the filtrate is concentrated in a vacuum to obtain a residue. The residue is purified by column chromatography to obtain [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, 17.2g) as a yellow oil. MS calculated value 758.3 (MH+ ); measured value 758.3 (MH + ).

[0430] 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)

[0431] To a solution 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, 15 g, 19.77 mmol) in DMF (300 mL) was added CsCO (19.3 g, 59.3 mmol) and iodoethane (6.16 g, 39.53 mmol) at 0°C. After stirring at 20°C for 16 hours, the reaction mixture was poured into water (200 mL) and extracted with EtOAc (200 mL, three times). The combined organic layers were washed with brine (10 mL, three times), dried over NaSO, 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-pyridinyl]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 + ).

[0432] 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)

[0433] To a solution 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, 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 hours. 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 three times with EtOAc (200 mL). The combined organic layer was washed with brine (200 mL, four times), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by column chromatography to give 3-[5-bromo-1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]indol-3-yl]-2,2-dimethyl-propan-1-ol as a colorless foam (Compound G5, 6 g, eluted faster) and 3-[5-bromo-1-ethyl-6-fluoro-(2P)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]indol-3-yl]-2,2-dimethyl-propan-1-ol as a colorless foam (Compound G6, 4.5 g, eluted slower). Compound G5: MS calculated value 548.2 (MH + ); measured value 548.2 (MH + ). 1 HNMR (400MHz, methanol-d4) δ = 8.41 (d, J = 2.4Hz, 1H), 7.92 (d, J = 6.8Hz, 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,1 H), 1.42 (d, J = 6.4Hz, 3H), 1.22 (t, J = 7.2Hz, 3H), 0.76 (s, 3H), 0.76 (s, 3H).

[0434] X-ray crystallography analysis of compound G5

[0435] The absolute configuration structure of compound G5 was confirmed by X-ray crystallography of its single crystal. Figure 1 ).

[0436] Step 6: Preparation of 3-[1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethyl-propan-1-ol (Compound G7)

[0437] To a 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), bis(pinacol)boronate (4.2 g, 16.41 mmol) in toluene (60 mL) was added potassium acetate (2.7 g, 27.35 mmol) and Pd(dppf)Cl2 (0.8 g, 1.09 mmol). The reaction mixture was degassed by bubbling nitrogen for 5 min 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 chromatography to give 3-[1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-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 + ).

[0438] Step 7: 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 methyl ester (Compound G8)

[0439] To 3-[1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-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 (3S)-1-[(2S)-3-(4-bromo To a mixture of toluene (45 mL), 1,4-dioxane (15 mL) and 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 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. The residue was purified by column chromatography 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-pyridinyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid methyl ester (Compound G8, 3.8 g) as a colorless gum. MS calculated value 866.4 (MH + ); measured value 866.4 (MH + ).

[0440] 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]hexahydropyrazine-3-carboxylic acid (Compound G9)

[0441] To a mixture 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]hexahydropyrazine-3-carboxylic acid methyl ester (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 hours. The reaction mixture was concentrated under vacuum to give a residue. EtOAc (200 mL) and water (100 mL) were added to the residue, and the layers were separated. The aqueous phase was extracted with EtOAc (150 mL, twice). The combined organic layers 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-pyridinyl]indol-5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (Compound G9, 3.7 g) as a brown solid. MS calculated value 852.4 (MH + ); measured value 852.4 (MH + ).

[0442] Step 9: Preparation of N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 tert-Butyl]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (Compound G10)

[0443] To a mixture 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]hexahydropyrazine-3-carboxylic acid (compound G9, 2.5 g, 2.93 mmol) in DCM (250 mL) was added DIEA (7.58 mL, 58.68 mmol), EDCI (8.4 g, 44.01 mmol) and HOBt (991.2 mg, 0.91 mmol) at 0° C. After stirring at 20° C. for 12 hours, the reaction mixture was poured into water (100 mL) and extracted with EtOAc (100 mL, three times). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue that was purified by column chromatography to afford N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow oil. 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamic acid tert-butyl ester (Compound G10, 1.2 g). MS calculated value 834.4 (MH + ); measured value 834.4 (MH + ).

[0444] Step 10: Preparation of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G)

[0445] To N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 To a solution of tert-butyl]octadecanoate-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). The mixture was stirred at 20 ° C for 3 hours. After the reaction was completed, the reaction mixture was concentrated under vacuum to give a residue. Saturated NaHCO3 aqueous solution (60 mL) was added and the mixture was extracted with EtOAc (80 mL, three times). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a yellow solid. 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G, 1 g). MS calculated value 734.3 (MH + ); measured value 734.3 (MH + ).

[0446] Intermediate H

[0447] (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0448]

[0449] The title compound was prepared in analogy to the preparation of intermediate C by using 5-bromo-4-fluoro-1H-indole instead of 5-bromo-6-fluoro-1H-indole (compound C2).

[0450] Intermediate I

[0451] (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0452]

[0453] Analogously to the preparation of intermediate E, the title compound was prepared by using morpholine instead of 1-(2,2,2-trifluoroethyl)piperazine (Compound E1).

[0454] Intermediate J

[0455] (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]Octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione

[0456]

[0457] Analogously to the preparation of intermediate E, the title compound was prepared by using [3-(5-bromo-4-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound J1) instead of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (Compound C5).

[0458] Analogously to the preparation of intermediate C5, compound J1 was prepared by using 5-bromo-4-fluoro-1H-indole instead of 5-bromo-6-fluoro-1H-indole (compound C2).

[0459] Example 1

[0460] 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide

[0461]

[0462] The compounds were prepared according to the following scheme:

[0463]

[0464] Step 1: Preparation of (2S)-2-[tert-butoxycarbonyl(methyl)amino]-3-methyl-butyric acid benzyl ester (Compound 1B)

[0465] To a mixture of BOC-N-ME-VAL-OH (5.0 g, 21.62 mmol) and potassium carbonate (4.5 g, 32.43 mmol) in DMF (70 mL) was added benzyl bromide (4.1 g, 23.78 mmol) at 0 ° C. The mixture was stirred at 25 ° C for 12 hours. After the reaction was completed, the reaction mixture was diluted with water (70 mL) and extracted with EtOAc (70 mL, twice). The combined organic layer was washed with brine (200 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by reverse phase column to obtain (2S)-2-[tert-butoxycarbonyl (methyl) amino]-3-methyl-butyric acid benzyl ester (Compound 1B, 6.7 g) as a yellow oil. 1H NMR (400 MHz, chloroform-d) δ = 7.34 (br s, 5H), 5.21-5.10 (m, 2H), 2.88-2.74 (m, 3H), 2.31-2.12 (m, 1H), 1.68-1.64 (m, 1H), 1.44 (br d, J = 15.0 Hz, 9H), 0.96 (d, J = 6.6 Hz, 3H), 0.90 (br d, J = 3.8 Hz, 3H). MS calculated value 322.2 (MH + ), measured value 344.2 (MNa + ).

[0466] Step 2: Preparation of (2S)-3-methyl-2-(methylamino)butyric acid benzyl ester (Compound 1C)

[0467] A mixture of (2S)-2-[tert-butoxycarbonyl(methyl)amino]-3-methyl-butyric acid benzyl ester (Compound 1B, 6.7 g, 21.47 mmol) in HCl / 1,4-dioxane (50.0 mL, 2M) was stirred at 25 °C for 1 h. After the reaction was complete, the reaction mixture was concentrated in vacuo to give a residue. The residue was diluted with saturated NaHCO3 (40 mL) and extracted with EtOAc (50 mL, three times). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated in vacuo to give (2S)-3-methyl-2-(methylamino)butyric acid benzyl ester (Compound 1C, 4.7 g) as a colorless oil. MS calculated value 222.1 (MH + ); measured value 222.2 (MH + ).

[0468] Step 3: Preparation of (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butyric acid benzyl ester (Compound 1D)

[0469] To a mixture of (2S)-3-methyl-2-(methylamino)butyric acid benzyl ester (compound 1C, 1.0 g, 4.52 mmol) and DIEA (0.9 mL, 4.97 mmol) in DCM (15 mL) was added triphosgene (1.3 g, 4.52 mmol) at 0 ° C. After stirring for 0.5 h, a solution of N, N-dimethylazetidinyl-3-amine dihydrochloride (compound 1D, 782.1 mg, 4.52 mmol) and DIEA (1.97 mL, 11.3 mmol) in DCM (4 mL) was added to the reaction. The reaction mixture was stirred for another 1 h at 20 ° C. After the reaction was completed, the reaction mixture was quenched with water (40 mL) and extracted with EtOAc (40 mL, twice). The combined organic layer was washed with brine (60 mL), dried over Na2SO4, filtered and concentrated in vacuo to obtain a residue. The residue was purified by preparative HPLC to give (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butyric acid benzyl ester (Compound IE, 1.0 g) as a yellow oil. MS calculated 348.2 (MH + ); measured value 348.2 (MH + ).

[0470] Step 4: Preparation of (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butyric acid (Compound 1F)

[0471] To a mixture of (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butyric acid benzyl ester (Compound 1E, 500.0 mg, 1.44 mmol) in methanol (15 mL) was added activated carbon-supported Pd(OH)2 (400.0 mg, 0.57 mmol). The reaction mixture was degassed and purged with hydrogen three times and stirred at 20 ° C for 12 hours under a hydrogen atmosphere (15 psi). After the reaction was complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butyric acid (Compound 1F, 370.0 mg) as a yellow oil. MS calculated value 258.1 (MH + ); measured value 258.3 (MH + ).

[0472] Step 5: Preparation of 3-(dimethylamino)-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22 ,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide (Example 1)

[0473] To a solution of (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butanoic acid (Compound 1F, 32.13 mg, 0.12 mmol) in DMF (2 mL) at 0 °C was added DIEA (0.1 mL, 0.25 mmol), HATU (47.47 mg, 0.12 mmol) and (7S,13S)-7-amino-24-fluoro-17,17-dimethyl-20-[5-(4-methylpiperazin-1-yl)-2-[rac-(1S)-1-methoxyethyl]-3-pyridinyl]-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 ] 28-hexadecane-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C, 50.0 mg, 0.06 mmol). After stirring at 20 °C for 1 h, the reaction mixture was diluted with water (15 mL) and extracted with EtOAc (15 mL, three times). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC to give Example 1 (35.1 mg) as a white solid. MS calculated value 1040.5 (MH + ), measured value 1040.8(MH + ), 1H NMR (400MHz, methanol-d4) δ = 8.68 (d, J = 7.6Hz, 1H), 8.51 (d, J = 2.8Hz, 1H), 7.69 (d, J = 2.4Hz, 1H), 7.57-7.44 (m, 2H), 5.79-5.69 (m, 1H), 5.25 -5.11(m,1H),4.97-4.89(m,2H),4.51-4.39(m,2H),4.31-4.12(m,7H),4.12-3.88(m,2H),3.87-3.67(m,3H),3.67-3.52(m,2H),3.47( d,J=14.8Hz,2H),3.35(s,3H),3.29-3.22(m,2H),3.22-3.10(m,2H),2.99(s,3H),2.92(s,8H),2.87-2.77(m,1H),2.58(d,J=14.4Hz,1 H),2.30-2.16(m,2H),2.03-1.90(m,1H),1.88-1.74(m,1H),1.72-1.58(m,1H),1.45(d,J=6.0Hz,3H),1.03-0.89(m,9H),0.45(s,3H).

[0474] Example 2

[0475] 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide

[0476]

[0477] The title compound was prepared similarly to Example 1 by using (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate H) was substituted with (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22 ,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 2 (50.0 mg) was obtained as a yellow solid. MS calculated value 1040.5 (MH + ); measured value 1040.8 (MH + ). 1 H NMR (400 MHz, methanol-d4) δ=8.55-8.49 (m, 1H), 7.63-7.56 (m, 1H), 7.52-7.39 (m, 3H), 6.03-5.81 (m, 1H), 5.20-5.01 (m, 1H), 4.70-4.61 (m, 1H), 4.45-4.36 (m, 2H), 4.30-4.23 (m, 2H), 4.17-3.93 (m, 7H), 3.77-3.63 (m, 3H), 3.56-3.48 (m, 2H), 3.42 -3.34(m,2H),3.29-3.21(m,3H),3.20-3.11(m,3H),3.00-2.98(m,3H),2.94-2.87(m,10H),2.68-2.51(m,2H),2.23-2. 14(m,1H),1.68-1.53(m,1H),1.47-1.41(m,3H),1.35-1.15(m,3H),0.95-0.88(m,6H),0.81(s,3H),0.72-0.58(m,3H).

[0478] Example 3

[0479] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide

[0480]

[0481] The title compound was prepared similarly to the preparation of Example 1 by using morpholine instead of N,N-dimethylazetidine-3-amine dihydrochloride (Compound 1D). Example 3 (25 mg) was obtained as an off-white solid. MS calculated value 1027.4 (MH + ), measured value 1027.7 (MH + ), 1 H NMR (400MHz, methanol-d4) δ = 8.71 (d, J = 7.6Hz, 1H), 8.49 (d, J = 2.8Hz, 1H), 7.72-7.67 (m, 2H), 7.48 (d, J = 12.8Hz, 1H), 5.76 (d, J = 8.8Hz, 1H), 5.26- 5.13(m,1H),4.84-4.75(m,1H),4.43(d,J=12Hz,1H),4.30-4.24(m,1H),4.20(dd,J=12,2.8Hz,1H),4.17-4.03(m,2H),4.00(s,1H),3.85-3. 62(m,8H),3.55-3.44(m,2H),3.42-3.34(m,6H),3.30-3.18(m,5H),3. 15-3.09(m,1H),2.99(s,3H),2.91(s,3H),2.87-2.77(m,1H),2.71-2. 58(m,1H),2.33-2.14(m,2H),2.01-1.90(m,1H),1.87-1.74(m,1H),17 0-1.57(m,1H),1.46(d,J=6.0Hz,3H),1.01-0.87(m,9H),0.50(s,3H).

[0482] Example 4

[0483] 3-(Dimethylamino)-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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide

[0484]

[0485] The title compound was prepared similarly to Example 1 by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate I) was substituted with (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 4 (12.9 mg) was obtained as a yellow solid. MS calculated value 1027.5 (MH + ); measured value 1027.5 (MH + ). 1H NMR (400MHz, methanol-d4) δ = 8.69 (d, J = 7.8Hz, 1H), 8.42 (d, J = 2.9Hz, 1H), 7.69 (d, J = 2.4Hz, 1H), 7.55 (d, J = 2.4Hz, 1H), 7.48 (d, J = 12.7Hz, 1H), 5.74 (br d,J=9.3Hz,1H),5.18(br dd,J=8.3,16.6Hz,1H),4.87-4.79(m,2H),4.49-4.38(m,2H),4.33-4.11(m,7H ),3.87(t,J=4.6Hz,4H),3.78(brd,J=11.2Hz,1H),3.72-3.66(m,1H),3.48(br d,J=14.7Hz,1H),3.39-3.33(m,6H),3.26-3.21(m,1H),3.15-3.10(m,1H),2.97-2.90(m,8H),2.89-2.76(m,2H),2.66-2.60(m,1H) ),2.26-2.17(m,2H),2.01-1.91(m,1H),1.87-1.78(m,1H),1.70-1.60(m,1H),1.49-1.41(m,3H),1.01-0.84(m,9H),0.48(s,3H).

[0486] Example 5

[0487] 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide

[0488]

[0489] The title compound was prepared similarly to Example 1 by using morpholino and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.12,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 5 (131.3 mg) was obtained as a yellow solid. MS calculated value 960.5 (MH + ); measured value 960.3 (MH + ). 1 H NMR (400MHz, chloroform-d) δ = 8.97 (br d,J=1.6Hz,1H),8.69(d,J=7.5Hz,1H),8.14-7.98(m,1H),7.61(d,J=1.7Hz,1H),7.38(d,J=1.5Hz,1H),7.13(d,J=12.5Hz,1H),5.81(br d,J=17.7Hz,1H),4.68-4.57(m,1H),4.41(d,J=6.5Hz,1H),4.29-4.22(m,1H) ,4.19-4.07(m,2H),4.00-3.88(m,6H),3.84-3.78(m,2H),3.77-3.70(m,3H), 3.49-3.42(m,6H),3.39-3.31(m,6H),3.23-3.14(m,2H),2.91(s,3H),2.77-2 .68(m,1H),2.49-2.42(m,1H),2.38-2.28(m,2H),2.02-1.96(m,2H),1.82(br d,J=12.7Hz,1H),1.69-1.60(m,1H),1.57(d,J=6.2Hz,3H),1.03-0.96(m,9H),0.91(d,J=6.6Hz,3H),0.55-0.43(m,3H).

[0490] Example 6

[0491] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholino-4-carboxamide

[0492]

[0493] The title compound was prepared similarly to Example 1 by using morpholine and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2 ,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 6 (8.8 mg) was obtained as an off-white solid. MS calculated value 1095.5 (MH + ); measured value 1095.5 (MH + ). 1H NMR (400MHz, methanol-d4) δ = 8.71 (d, J = 7.2Hz, 1H), 8.45-8.38 (m, 1H), 7.69 (d, J = 2.0Hz, 1H), 7.63-7.56 (m, 1H), 7.48 (d, J = 12 .8Hz,1H),5.87-5.69(m,1H),5.24-5.11(m,1H),4.49-4.37(m,1H),4.29-4.16(m,2H),4.01(d,J=11.2Hz,1H),3.83-3. 57(m,7H),3.50-3.36(m,10H),3.26-3.06(m,5H),2.93-2.77(m,8H),2.71-2.60(m,1H),2.31-2.16(m,2H),2.00-1.91( m,1H),1.87-1.75(m,1H),1.70-1.58(m,1H),1.46(d,J=6.0Hz,3H),1.34-1.26(m,1H),1.00-0.88(m,9H),0.50(s,3H).

[0494] Example 7

[0495] 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-3,3-difluoro-N-methyl-azetidine-1-carboxamide

[0496]

[0497] The compounds were prepared according to the following scheme:

[0498]

[0499] Step 1: N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .19,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamic acid tert-butyl ester (Compound 7A)

[0500] To a solution of BOC-N-ME-VAL-OH (Compound 1A, 66.2 mg, 0.29 mmol) in DMF (1 mL) was added DIEA (0.3 mL, 1.90 mmol) and HATU (67.3 mg, 0.29 mmol) at 0° C. After stirring at 0° C. for 15 min, (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 2,5 .1 9,13 .0 22 ,26 ] 28-hexadecane-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G, 70.0 mg, 0.09 mmol). After the reaction was stirred at 20 °C for 0.5 h, the reaction mixture was poured into water (10 mL) and extracted with ethyl acetate (10 mL, three times). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by preparative TLC to give N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a light yellow solid. 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamic acid tert-butyl ester (Compound 7A, 65.1 mg). MS calculated value 947.5 (MH + ); measured value 947.4 (MH + ).

[0501] Step 2: [(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-methyl-ammonium; 2,2,2-trifluoroacetate (Compound 7B)

[0502] At 20°C, N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 To a mixture of 1-octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamic acid tert-butyl ester (Compound 7A, 40.0 mg, 0.04 mmol) in DCM (1 mL) was added TFA (0.2 mL, 2.7 mmol). After stirring at 20° C. for 1 h, the reaction mixture was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC to give [(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1] as a white solid. 2,5 .1 9 ,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-methyl-ammonium; 2,2,2-trifluoroacetate (Compound 7B, 20.0 mg). MS calculated value 847.5 (MH + ), measured value 847.4 (MH + ). 1H NMR (400MHz, methanol-d4) δ = 8.68 (d, J = 7.8Hz, 1H), 8.40 (d, J = 2.8Hz, 1H), 7.65 (d, J = 2.4Hz, 1H), 7.56 (br s,1H),7.35(d,J=12.6Hz,1H),5.95-5.92(m,1H),5.04-4.96(m,1H),4.94-4.91(m,3H),4.45

[0503] -4.40(m,1H),4.35-4.05(m,4H),3.88-3.83(m,4H),3.80-3.69(m,3H),3.56

[0504] -3.50(d,J=14.5Hz,1H),3.35-3.31(m,4H),3.07-2.97(m,1H),2.90-2.80(m,1H),2.75-2.65(m,4H),2.30-2.15(m,2H),2.03-1.91(m,1H), 1.89-1.75(m,1H),1.73-1.56(m,1H),1.45-1.43(m,3H),1.15-1.09(m ,3H),1.09-1.07(m,3H),1.03-1.01(m,3H),0.95(s,3H),0.56(s,3H).

[0505] Step 3: N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-3,3-difluoro-N-methyl-azetidine-1-carboxamide (Example 7)

[0506] At 0°C, [(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26] 2-octadecane-1 (25), 2,5 (28), 19,22 (26), 23-hexaen-7-yl] carbamoyl] -2-methyl-propyl] -methyl-ammonium; 2,2,2-trifluoroacetate (compound 7B, 50.0 mg, 0.06 mmol) and DIEA (45.8 mg, 0.35 mmol) in DCM (1 mL) solution was added triphosgene (6.1 mg, 0.02 mmol). After stirring at 20 ° C for 1 h, 3,3-difluoroazetidine hydrochloride (compound 7C, 7.6 mg, 0.06 mmol) was added to the reaction mixture at 0 ° C and stirred at 20 ° C for 11 hours. After the reaction was completed, the reaction mixture was poured into H2O (10 mL) and extracted with EtOAc (10 mL, three times). The organic layer was washed with brine (10 mL, twice), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by preparative HPLC to give Example 7 (13.0 mg) as a light yellow solid. MS calculated 966.5 (MH + ); measured value 966.4 (MH + ). 1 HNMR (400 MHz, methanol-d4) δ = 8.70-8.64 (m, 1H), 8.49-8.35 (m, 1H), 7.62 (d, J = 2.2 Hz, 1H), 7.51 (d, J = 2.4 Hz, 1H), 7.33 (d, J = 12.8 Hz, 1H), 5.78 (br t,J=8.2Hz,1H),5.02-4.96(m,1H),4.68-4.52(m,1H),4.46-4.43(m,1H),4.42-4.32(m,4H),4.22-4.0 5(m,3H),3.93-3.82(m,4H),3.81-3.66(m,2H),3.50-3.42(m,1H),3.35-3.31(m,6H),3.28-3.21(m,1H ),3.09-2.96(m,1H),2.88(s,3H),2.84-2.77(m,1H),2.26-2.14(m,2H),1.95-1.90(m,1H),1.87-1.74 (m,1H),1.69-1.59(m,1H),1.43(d,J=6.2Hz,3H),1.36-1.27(m,2H),1.03-0.89(m,12H),0.53(s,3H).

[0507] Example 8

[0508] (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butanamide

[0509]

[0510] The title compound was prepared similarly to Example 1 by using (2S)-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butanoic acid (Compound 8H) 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-tetraazapentacyclo-[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was substituted with (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butyric acid (Compound 1F) 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 8 (50.4 mg) was obtained as a yellow solid. MS calculated value 916.5 (MH + ), measured value 916.3 (MH + ). 1H NMR (400MHz, chloroform-d) δ = 8.84 (br s,1H),8.66(d,J=7.6Hz,1H),7.58(d,J=2.0Hz,1H),7.10(d,J=12.3Hz,1H),7.05(d,J=9.2Hz,1H),5.83(t,J=9.0Hz,1H),4.59(br d,J=11.9Hz,1H),4.39-4.31(m,1H),4.27-4.14(m,2H),4.11-4.03(m,1H),3.95(s,1H),3.93-3.89(m,4H),3.84(br d,J=11.5Hz,1H),3.71(d,J=11.0Hz,1H),3.52-3.46(m,1H),3.42-3.31(m,11H),3.21-3.07(m,3H),2.89(s,3H),2.76-2.59(m,2H),2.46(br d,J=14.4Hz,1H),2.29-2.18(m,2H),2.00-1.91(m,1H),1.80(br d,J=12.2Hz,1H),1.66-1.59(m,1H),1.54-1.50(m,3H),1.00-0.90(m,12H),0.49(s,3H).

[0511] Compound 8H was prepared according to the following scheme:

[0512]

[0513] Step 1: Preparation of (2S)-2-(tert-butoxycarbonylamino)-3-methyl-butyric acid benzyl ester (Compound 8B)

[0514] To a mixture of BOC-L-valine (Compound 8A, 1.0 g, 4.6 mmol) and potassium carbonate (763.4 mg, 5.52 mmol) in DMF (10 mL) was added benzyl bromide (0.6 mL, 5.06 mmol) at 0 ° C. The mixture was stirred at 25 ° C for 12 hours. After the reaction was completed, the reaction mixture was diluted with water (70 mL) and extracted with EtOAc (70 mL, three times). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by column chromatography to obtain (2S)-2-(tert-butoxycarbonylamino)-3-methyl-butyric acid benzyl ester (Compound 8B, 1.0 g) as a colorless oil. MS calculated value 307.4 (MH + ), measured value 208.0 (M-Boc+H + ).

[0515] Step 2: Preparation of (2S)-2-amino-3-methyl-butyric acid benzyl ester (Compound 8C)

[0516] To a mixture of (2S)-2-(tert-butoxycarbonylamino)-3-methyl-butyric acid benzyl ester (Compound 8B, 1.0 g, 3.11 mmol) in THF (5 mL) was added HCl / dioxane (20.0 mL, 80.0 mmol, 4 N) at 0°C. After the reaction was stirred at 25°C for 1 h, the reaction mixture was concentrated under vacuum to give a yellow gum. The gum was diluted with aqueous NaHCO3 solution (40 mL) and extracted with EtOAc (50 mL, three times). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under vacuum to give (2S)-2-amino-3-methyl-butyric acid benzyl ester (Compound 8C, 600.0 mg) as a colorless oil, which was used in the next step without purification.

[0517] Step 3: Preparation of (2S)-2-[2-[tert-Butoxycarbonyl(methyl)amino]-ethylamino]-3-methyl-butyric acid benzyl ester (Compound 8E)

[0518] To a solution of (2S)-2-amino-3-methyl-butyric acid benzyl ester (Compound 8C, 600.0 mg, 2.89 mmol) and N-BOC-(methylamino)acetaldehyde (Compound 8D, 601.7 mg, 3.47 mmol) in methanol (20 mL) was added MgSO4 (2.1 g, 14.47 mmol). After stirring at 20°C for 1 h, NaBH(OAc)3 (1.2 g, 5.79 mmol) was added to the reaction mixture and stirred at 20°C for another 1 h. After the reaction was complete, the reaction mixture was filtered and the filtrate was concentrated under vacuum to give a colorless oil. The oil was purified by column chromatography to give (2S)-2-[2-[tert-butoxycarbonyl(methyl)amino]-ethylamino]-3-methyl-butyric acid benzyl ester (Compound 8E, 700.0 mg) as a colorless gum. MS calculated value 365.2 (MH + ); measured value 365.1 (MH + ).

[0519] Step 4: Preparation of (2S)-3-methyl-2-[2-(methylamino)ethylamino]butyric acid benzyl ester; hydrochloride (Compound 8F)

[0520] To a solution of (2S)-2-[2-[tert-butoxycarbonyl(methyl)amino]-ethylamino]-3-methyl-butyric acid benzyl ester (Compound 8E, 700.0 mg, 1.92 mmol) in THF (5 mL) was added HCl / dioxane (15.0 mL, 60.0 mmol, 4N) at 0°C. After stirring at 25°C for 2 hours, the reaction mixture was concentrated under vacuum to give a white solid. The residual solvent was removed twice by azeotropic distillation with THF to give (2S)-3-methyl-2-[2-(methylamino)ethylamino]butyric acid benzyl ester; hydrochloride (Compound 8F, 600.0 mg) as a white solid. It was used directly in the next step without further purification. MS calculated value 265.2 (MH + ), measured value 265.2(MH + ).

[0521] Step 5: Preparation of (2S)-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butyric acid benzyl ester (Compound 8G)

[0522] To a solution of (2S)-3-methyl-2-[2-(methylamino)ethylamino]butyric acid benzyl ester; hydrochloride (compound 8F, 600.0 mg, 1.99 mmol) and DIEA (772.5 mg, 5.98 mmol) in ACN (50 mL) was added CDI (646.4 mg, 3.99 mmol). After stirring at 0 ° C for 2 hours, water (10 mL) was added to the reaction mixture and extracted with EA (40 mL, twice). The combined organic layers were washed with brine (20 mL), dried (Na2SO4), and concentrated under vacuum to give a yellow gum. The gum was purified by reverse phase column to give (2S)-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butyric acid benzyl ester (compound 8G, 300.0 mg) as a colorless gum. MS calculated value 291.2 (MH + ); measured value 291.2 (MH + ).

[0523] Step 6: Preparation of (2S)-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butanoic acid (Compound 8H)

[0524] To a solution of (2S)-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butyric acid benzyl ester (Compound 8G, 280.0 mg, 0.96 mmol) in methanol (10 mL) was added Pd(OH)2 (0.2 g, 10% purity) on activated carbon. The reaction mixture was hydrogenated at room temperature for 5 hours. The reaction mixture was filtered and the filtrate was concentrated under vacuum to give (2S)-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butyric acid (Compound 8H, 150.0 mg) as a colorless gum. MS calculated value 201.1 (MH + ); measured value 201.0 (MH + ).

[0525] Example 9

[0526] 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-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide

[0527]

[0528] The title compound was prepared similarly to Example 1 by using morpholino 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-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate F) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 9 (26.8 mg) was obtained as an off-white solid. MS calculated value 1041.5 (MH + ); measured value 1041.7 (MH + ). 1 H NMR (400MHz, methanol-d4) δ=8.71(d,J=7.6Hz,1H),8.38(d,J=2.8Hz,1H),7.87(d,J=2.8Hz,1H),7.64(d,J=2.4Hz,1H),7.36(d,J=12.8Hz ,1H),5.86(d,J=8.4Hz,1H),4.48-4.32(m,2H),4.30-4.19(m,1H),4.18-4.11(m,1H),4.08-3.97(m,2H),3.80-3.67(m,6H),3.51-3. 47(m,4H),3.39-3.33(m,5H),3.29-3.08(m,6H),3.03-2.96(m,1H),2.93-2.84(m,7H),2.84-2.70(m,2H),2.28-2.14(m,2H),1.99-1 .92(m,1H),1.86-1.74(m,1H),1.70-1.58(m,1H),1.48-1.43(d,J=6.0Hz,3H)1.03(t,J=6.8Hz,3H),0.96-0.89(m,9H),0.61(s,3H).

[0529] Example 10

[0530] N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide

[0531]

[0532] The title compound was prepared similarly to Example 1 by using morpholino and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 10 (10.3 mg) was obtained as a yellow solid. MS calculated value 973.5 (MH + ); measured value 973.5 (MH + ). 1H NMR (400MHz, methanol-d4) δ=8.67(d,J=7.6Hz,1H),8.49(d,J=2.8Hz,1H),7.70-7.60(m,2H),7.34(d,J=12.4Hz,1H),5.80(d,J=8.8Hz ,1H),4.43(d,J=10.8Hz,1H),4.36-4.28(m,1H),4.27-4.05(m,4H),4.01(d,J=10.8Hz,1H),3.81-3.62(m,8H),3.49-3.43(m,2H ),3.42-3.33(m,7H),3.29-3.17(m,4H),3.07-3.02(m,1H),2.99(s,3H),2.89(s,3H),2.86-2.79(m,1H),2.67-2.63(m,1H),2.3 0-2.15(m,2H),2.01-1.91(m,1H),1.86-1.74(m,1H),1.70-1.57m,1H),1.44(d,J=6.0Hz,3H),1.03-0.88(m,13H),0.53(s,3H).

[0533] Example 11

[0534] N-[(1S)-1-[[(7S,13S)-24-fluoro-20-(20M)-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholino-4-carboxamide

[0535]

[0536] The title compound was prepared similarly to Example 1 by using morpholino and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate I) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 11 (26.3 mg) was obtained as a white solid. MS calculated value 1013.5 (MH + ); measured value 1013.8 (MH + ). 1 H NMR (400MHz, methanol-d4) δ = 8.71 (d, J = 7.6 Hz, 1H), 8.49-8.35 (d, J = 2.8, 1H), 7 .73-7.69(d,J=2.4,1H),7.66-7.59(d,J=2.4,1H),7.53-7.43(d,J=12.4 ,1H),5.88-5.70(d,J=8.8,1H),5.25-5.13(m,1H),4.53-4.38(m,1H),4. 31-4.15(m,2H),4.07-3.98(d,J=10.8,1H),3.91-3.84(t,J=4.8,4H),3.8 3-3.67(m,6H),3.52-3.45(m,1H),3.43-3.34(m,10H),3.30-3.18(m,3H) ,3.11(m,1H),2.99-2.89(s,3H),2.88-2.78(m,1H),2.72-2.60(d,J=14.8 ,1H),2.34-2.15(m,2H),2.01-1.91(m,1H),1.90-1.74(m,1H),1.72-1.5 7(m,1H),1.52-1.42(d,J=6.0,3H),1.05-0.88(m,9H),0.57-0.45(s,3H).

[0537] Example 12

[0538] 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide

[0539]

[0540] The title compound was prepared similarly to Example 1 by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9, 13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E) was substituted with (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octadecyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 12 (45.3 mg) was obtained as a white solid. MS calculated value 1108.5 (MH + ); measured value 1108.5 (MH + ). 1H NMR (400MHz, methanol-d4) δ = 8.68 (d, J = 7.6Hz, 1H), 8.42 (d, J = 2.8Hz, 1H), 7.70 (d, J = 2.4Hz, 1H), 7.45 (d, J = 12.8Hz, 1H), 7.32 -7.27(m,1H),5.74-5.68(m,1H),5.18-5.06(m,2H),4.93-5.00(m,2H),4.58-4.56(m,5H),4. 32-4.23(m,1H),4.22-4.14(m,2H),4.14-4.07(m,1H),4.00-3.89(m,1H),3.87-3.67(m,2H),3 .53-3.39(m,1H),3.34-3.32(m,4H),3.19-3.09(m,4H),2.90-2.79(m,8H),2.63-2.54(m,1H) ,2.26-2.15(m,8H),1.57-1.98(m,3H),1.43(d,J=6.0Hz,3H),0.99-0.89(m,9H),0.44(s,3H).

[0541] Example 13

[0542] (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-[methyl(2,2,2-trifluoroethylcarbamoyl)amino]butanamide

[0543]

[0544] The title compound was prepared similarly to Example 1 by using 2,2,2-trifluoroethylamine and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 2,5 .1 9,13 .0 22,26] octadecyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 13 (13.4 mg) was obtained as a white solid. MS calculated value 972.4 (MH + ); measured value 972.4 (MH + ). 1 H NMR (400MHz, chloroform-d) δ = 8.64 (d, J = 7.6Hz, 1H), 8.47 (d, J = 2.8Hz, 1H), 7.55 (d, J =1.7Hz,1H),7.10(s,1H),7.08-7.03(m,2H),5.84(t,J=9.1Hz,1H),5.21(br s,1H),4.58(br d,J=12.1Hz,1H),4.32-4.21(m,3H),4.08-4.02(m,2H),3.92-3.88(m,4H),3.84(br d,J=11.0Hz,1H),3.73-3.70(m,1H),3.38(s,3H),3.27-3.21(m,4H),3.17-3.04(m,2H),2.97(s,3H),2.74-2.65(m,1H),2.46(br d,J=14.4Hz,1H),2.31-2.21(m,5H),1.96(br d,J=13.7Hz,1H),1.85-1.72(m,1H),1.66-1.54(m,1H),1.44(d,J=6.1Hz,3H),1.25(t,J=7.0Hz,1H),0.99-0.86(m,12H),0.45(s,3H).

[0545] Example 14

[0546] 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-(trifluoromethyl)azetidine-1-carboxamide

[0547]

[0548] The title compound was prepared similarly to Example 1 by using 3-(trifluoromethyl)azetidine and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was substituted with N,N-dimethylazetidin-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 14 (3.1 mg) was obtained as a white solid. MS calculated value 998.5 (MH + ); measured value 998.5 (MH + ). 1H NMR (400MHz, chloroform-d) δ = 8.64 (d, J = 7.5Hz, 1H), 8.47 (d, J = 2.4Hz, 1H), 7.58 (s, 1H), 7.32 (br d, J = 9.6Hz, 1H), 7.11-7.04 (m, 2H), 5.81 (br t,J=8.8Hz,1H),4.59(br d,J=9.8Hz,1H),4.35(br t,J=8.7Hz,1H),4.30-4.21(m,4H),4.13(br dd,J=7.9,15.0Hz,3H),4.08-3.99(m,3H),3.90(br s,4H),3.84(br d,J=11.3Hz,1H),3.43(br d,J=14.6Hz,1H),3.37(s,3H),3.24(br d,J=4.3Hz,4H),3.18-3.13(m,1H),2.84(s,3H),2.75-2.62(m,1H),2.47(br d,J=14.4Hz,1H),2.31-2.17(m,2H),2.05(s,3H),1.94(br d,J=1.8Hz,1H),1.51(br s,2H),1.46-1.46(m,1H),1.44-1.44(m,1H),1.15(td,J=6.4,13.0Hz,12H),0.46(s,3H).

[0549] Example 15

[0550] (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazin-2-yl)butanamide

[0551]

[0552] The title compound was prepared similarly to Example 1 by using (2S)-3-methyl-2-(3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazin-2-yl)butanoic acid (Compound 15g) 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-tetraazapentacyclo-[175.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) was substituted with (2S)-2-[[3-(dimethylamino)azetidine-1-carbonyl]-methyl-amino]-3-methyl-butyric acid (Compound 1F) 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 15 (37.5 mg) was obtained as a white solid. MS calculated value 958.5 (MH + ); measured value 958.5 (MH + ). 1H NMR (400MHz, methanol-d4)δ=8.70-8.61(m,1H),8.53-8.30(m,1H),7.64-7.59(m,1H),7.38-7.27(m,2H),5.86-5.72(m,1H),5.03-4.92(m,1H),4 .82-4.78(m,1H),4.67-4.53(m,3H),4.47-4.36(d,J=12.8Hz,1H),4.29-4.12(m,4H),4.10-4.01(d,J=10.8Hz,1H),3.89-3.84(m,4H),3.7 9-3.68(m,4H),3.65-3.54(m,1H),3.50-3.36(m,4H),3.28(m,4H),3. 20-3.10(m,2H),3.03-2.95(m,1H),2.88-2.75(m,1H),2.69-2.58(m, 1H),2.24-2.07(m,2H),2.01-1.89(m,1H),1.87-1.71(m,1H),1.70-1 .56(m,1H),1.51-1.37(m,3H),1.03-0.87(m,12H),0.62-0.42(m,3H).

[0553] Compound 15g was prepared according to the following scheme:

[0554]

[0555] Step 1: 3-[[(1S)-1-Benzyloxycarbonyl-2-methyl-propyl]carbamoyl]-morpholino-4-carboxylic acid tert-butyl ester (Compound 15c)

[0556] To a solution of 4-tert-butoxycarbonylmorpholine-3-carboxylic acid (compound 15b, 2.8 g, 12.31 mmol) and L-valine benzyl ester hydrochloride (compound 15a, 3.0 g, 12.31 mmol) in DMF (30 mL) was added DIEA (4.29 mL, 24.62 mmol) and HATU (5.1 g, 13.54 mmol). After stirring at 25 °C for 1 h, EtOAc (40 mL) and water (40 mL) were added to the reaction mixture. The layers were separated and the aqueous phase was extracted with EtOAc (30 mL, twice). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give tert-butyl 3-[[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]carbamoyl]morpholino-4-carboxylate (compound 15c, 7.5 g) as a white oil. MS calculated value 420.23 (MH +), measured value 321.2 (M-Boc+H + ).

[0557] Step 2: (2S)-3-Methyl-2-(morpholino-3-carbonylamino)-butyric acid benzyl ester; hydrochloride (Compound 15d)

[0558] To a solution of tert-butyl 3-[[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]carbamoyl]morpholine-4-carboxylate (compound 15c, 7.2 g, 17.16 mmol) in 1,4-dioxane (10 mL) was added 1,4-dioxane / HCl (4 M, 15.0 mL, 60.0 mmol). The mixture was stirred at 25 °C for 2 hours. The mixture was concentrated under vacuum to give (2S)-3-methyl-2-(morpholine-3-carbonylamino)-butyrate; hydrochloride (compound 15d, 6.7 g) as a white oil. MS calculated 321.2 (MH + ); measured value 321.2 (MH + ).

[0559] Step 3: (2S)-2-(1,3-dioxo-5,6,8,8a-tetrahydroimidazo[5,1-c][1,4]oxazin-2-yl)-3-methyl-butyric acid benzyl ester (Compound 15e)

[0560] To a solution of (2S)-3-methyl-2-(morpholino-3-carbonylamino)-butyric acid benzyl ester (compound 15d, 6.5 g, 20.29 mmol) in THF (1 mL) was added TEA (8.5 mL, 60.87 mmol) and N.N'-carbonyldiimidazole (3.9 g, 24.35 mmol). After stirring at 25 °C for 2 hours, the reaction mixture was concentrated under vacuum to give a residue. The residue was purified by reverse phase chromatography to give (2S)-2-(1,3-dioxo-5,6,8,8a-tetrahydroimidazo[5,1-c][1,4]oxazin-2-yl)-3-methyl-butyric acid benzyl ester (compound 15e, 2.36 g) as a white oil. MS calculated value 346.4 (MH + ), measured value 369.4 (MNa + ).

[0561] Step 4: (2S)-3-methyl-2-(1-oxo-5,6,8,8a-tetrahydro-3H-imidazo[5,1-c][1,4]oxazin-2-yl)butanoic acid benzyl ester (Compound 15e)

[0562] To a solution of (2S)-2-(1,3-dioxo-5,6,8,8a-tetrahydroimidazo[5,1-c][1,4]oxazine-2-yl)-3-methyl-butyric acid benzyl ester (compound 15e, 1 g, 2.89 mmol) in THF (5 mL) was added boron trifluoride etherate (1 M, 1.2 mL, 1.2 mmol) and borane-THF (5.77 mL, 5.77 mmol) at 0 ° C. After stirring at 20 ° C for 18 hours, the reaction was quenched with water (40 mL). The aqueous phase was extracted with EtOAc (30 mL, twice). The combined organic layers were washed with brine (60 mL), dried over Mg2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by reverse phase column to give benzyl (2S)-3-methyl-2-(1-oxo-5,6,8,8a-tetrahydro-3H-imidazo[5,1-c][1,4]oxazin-2-yl)butanoate (Compound 15f, 220.0 mg) as a colorless oil. MS calculated value 333.2 (MH + ); measured value 333.2 (MH + ).

[0563] Step 5: (2S)-3-methyl-2-(3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazin-2-yl)butanoic acid (Compound 15f)

[0564] To a solution of benzyl (2S)-3-methyl-2-(1-oxo-5,6,8,8a-tetrahydro-3H-imidazo[5,1-c][1,4]oxazin-2-yl)butanoate (compound 15e, 130.0 mg, 0.39 mmol) in THF (1 mL) was added Pd / C (40.0 mg, 0.78 mmol). The reaction mixture was stirred at 25 °C under a hydrogen atmosphere for 1 h. The mixture was filtered and the filtrate was concentrated under vacuum to give (2S)-3-methyl-2-(3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazin-2-yl)butanoic acid (compound 15g, 62.0 mg) as a colorless oil. MS calculated 243.1 (MH + ); measured value 243.2 (MH + ).

[0565] Example 16

[0566] (3R)-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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-3-hydroxy-N,3-dimethyl-piperidine-1-carboxamide

[0567]

[0568] The title compound was prepared similarly to Example 1 by using (3R)-3-methylpiperidin-3-ol; hydrochloride and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9 ,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate I) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 16 (13.9 mg) was obtained as a white solid. MS calculated value 1042.5 (MH + ); measured value 1042.5 (MH + ). 1H NMR (400 MHz, methanol-d4) δ = 8.73-8.69 (d, J = 7.2 Hz, 1H), 8.44-8.39 (d, J = 2.8 Hz, 1H), 7.70-7.67 (d, J = 2.8 Hz, 1H), 7.67-7.64 (d, J = 2.4 Hz, 1H), 7.51-7.45 (d, J = 12.4 Hz, 1H), 5.80-5.74 (d, J = 8.4 Hz, 1H), 5.23-5.13 (m,1H),4.83-4.76(m,1H),4.47-4.40(m,1H),4.29-4.23(q,J=6.4Hz,J=12.0Hz,1H),4.23-4.17(dd,J =2.4,11.6Hz,1H),4.05-3.99(d,J=10.8Hz,1H),3.89-3.85(t,J=4.4Hz,4H),3.82-3.78(d,J=11.2Hz,1 H),3.74-3.69(d,J=11.2Hz,1H),3.56-3.45(m,2H),3.39-3.36(m,6H),3.29-3.19(m,2H),3.14-3.06( m,1H),3.03-2.96(d,J=12.8Hz,2H),2.91-2.89(s,3H),2.87-2.78(m,1H),2.71-2.63(m,1H),2.29-2.1 6(m,2H),2.00-1.90(m,3H),1.88-1.76(m,1H),1.74-1.68(m,1H),1.67-1.53(m,3H),1.49-1.45(d,J=6 .4Hz,3H),1.34-1.27(m,1H),1.25-1.21(s,3H),0.99-0.93(dd,J=6.0,12.0Hz,9H),0.56-0.49(s,3H).

[0569] Example 17

[0570] N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholino-4-carboxamide

[0571]

[0572] The title compound was prepared similarly to Example 1 by using morpholino and (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2 ,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate J) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 17 (12.9 mg) was obtained as a yellow solid. MS calculated value 1094.5 (MH + ); measured value 1094.5 (MH + ). 1H NMR (400MHz, methanol-d4)δ=8.49-8.39(m,1H),7.74(br s,1H),7.53-7.44(m,3H),5.97(br d,J=5.9Hz,1H),5.18-5.11(m,1H),4.67-4.58(m,1H),4.42-4.33(m,1H),4.10-3.97(m,1H),3.94 -3.80(m,2H),3.76-3.63(m,5H),3.58-3.42(m,7H),3.26-3.14(m,8H),3.13-2.99(m,2H),2.93(br d,J=14.4Hz,8H),2.62-2.53(m,1H),2.30-2.14(m,1H),1.61-1.49(m,1H),1.43(br d,J=6.1Hz,3H),1.34-1.15(m,3H),0.92(br t,J=7.3Hz,6H),0.85-0.69(m,6H).

[0573] Example 18

[0574] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N,4-dimethyl-piperazine-1-carboxamide; and

[0575]

[0576] The title compound was prepared similarly to Example 1 by using 1-methylpiperazine and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22 ,26 ] octadecyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 18 (10.8 mg) was obtained as a white solid. MS calculated value 1108.5 (MH + ); measured value 1108.5 (MH + ). 1 H NMR (400MHz, methanol-d4) δ = 8.70-8.65 (d, J = 7.6Hz, 1H), 8.45-8.40 (d, J = 2.8Hz ,1H),7.70-7.65(d,J=2.4Hz,1H),7.50-7.43(d,J=12.8Hz,1H),7.33-7.28 (d,J=2.8Hz,1H),5.78-5.70(d,J=9.2Hz,1H),5.20-5.05(m,1H),4.62-4. 61(s,1H),4.46-4.40(m,1H),4.28-4.20(m,1H),4.19-4.13(m,1H),4.06-3 .99(d,J=11.2Hz,1H),3.83-3.65(m,3H),3.53-3.40(m,4H),3.27-3.07(m ,6H),2.94-2.81(m,9H),2.63-2.45(m,6H),2.37-2.34(s,3H),2.29-2.17( m,3H),2.00-1.93(m,1H),1.90-1.75(m,2H),1.71-1.56(m,1H),1.46-1.41 (d,J=6.4Hz,3H),1.33-1.15(m,1H),0.98-0.91(m,9H),0.47-0.42(s,3H).

[0577] Example 19

[0578] N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(2,2,2-trifluoroethyl)piperazine-1-carboxamide

[0579]

[0580] The title compound was prepared similarly to Example 1 by using 1-(2,2,2-trifluoroethyl)piperazine and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ] octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E) was substituted with N,N-dimethylazetidinyl-3-amine dihydrochloride (Compound 1D) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9, 13 .0 22,26 ] prepared from octacosyl-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C). Example 19 (12.1 mg) was obtained as a white solid. MS calculated value 1176.5 (MH + ); measured value 1176.5 (MH + ). 1H NMR (400 MHz, methanol-d4) δ = 8.82-8.63 (d, J = 7.2 Hz, 1H), 8.48-8.38 (d, J = 2.8 Hz, 1H), 7.75-7.67 (d, J = 2.0 Hz, 1H), 7.54-7.43 (d, J = 12.4 Hz, 1H), 7.35-7.28 (d, J = 2.4 Hz, 1H), 5.86-5.72 (d, J = 8.8 Hz, 1H),5.26-5.03(m,1H),4.98-4.90(m,3H),4.66-4.56(m,1H),4.51-4.41(m,1H),4.32-4.13(m, 2H),4.07-4.03(d,J=11.3Hz,1H),3.86-3.77(d,J=11.6Hz,1H),3.76-3.69(d,J=11.2Hz,1H),3 .57-3.48(m,1H),3.48-3.41(m,2H),3.39-3.35(d,J=3.1Hz,4H),3.29-3.23(m,1H),3.22-3.10 (m,6H),2.96-2.92(s,3H),2.91-2.86(t,J=4.8Hz,4H),2.85-2.76(m,3H),2.76-2.69(m,2H),2 .67-2.57(d,J=14.4Hz,1H),2.33-2.19(m,2H),2.03-1.94(m,1H),1.92-1.79(m,1H),1.73-1.5 9(m,1H),1.50-1.41(d,J=6.0Hz,3H),1.35-1.30(m,1H),1.00-0.92(m,9H),0.54-0.41(s,3H).

[0581] Biological Examples

[0582] Compound A555 from WO2021091956 (page 158 of Table 1) is cited as a reference compound of the present invention.

[0583]

[0585] Example 20

[0586] Single-dose pharmacokinetic (PK) study in female BALB / c mice

[0587] The purpose of this study is to determine the pharmacokinetics of the selected compound after a single intravenous injection or oral gavage administration in female BALB / c mice. In short, two groups of female BALB / c mice (available from Shanghai Lingchang Biotechnology Co., Ltd) (N=3 / group) were treated intravenously with a single dose of the compound at 3mg / kg (IV) or orally at 30mg / kg (PO). Blood samples were collected at 5min (only for IV), 15min, 30min, 1h, 2h, 4h, 7h and 24h after administration. The blood samples were placed on ice until centrifugation to obtain plasma samples. The concentration of the compound in the plasma sample was determined using the LC-MS / MS method. Pharmacokinetic parameters were calculated by non-compartmental analysis.

[0588] Table 1. Results of SDPK

[0589]

[0590] As can be seen from Table 1, Example 6 has excellent pharmacokinetic properties in the mouse model, including C 最大 Almost 7 times, AUC 0-最后 The bioavailability of the drug was improved by 15 times, and the clearance rate was significantly reduced.

[0591] Example 21

[0592] Human hepatocyte stability assay

[0593] The hepatocyte stability assay measures the rate of compound disappearance when incubated with cryopreserved human suspension hepatocytes. Positive controls, including midazolam, raloxifene, and dextromethorphan, were included in each experiment. 1 μM of the test compound was incubated with a suspension of human hepatocytes (1×10 6 Cells / mL) were composed in Williams'E medium supplemented with 10% FBS and 0.5% penicillin-streptomycin. The hepatocyte suspension was cultured in a 5% CO2 incubator at 37°C with intermittent shaking at 900 rpm. The reaction was terminated by adding methanol containing an internal standard (2 μM tolbutamide) 2, 10, 20, 40, 60 and 120 minutes after the addition of the compound, and the consumption of the parent compound was monitored by LC-MS / MS analysis.

[0594] Table 2. Human hepatocyte stability of examples and compounds of the present invention

[0595]

[0596] The above results clearly show that in the human hepatocyte stability assay, the reference compound (A555) has a much higher clearance rate, while Example 6 maintains a low clearance rate. Achieving a low clearance rate is beneficial for improving the in vivo performance of a compound, such as dose reduction, exposure enhancement, and half-life extension.

[0597] Example 22

[0598] Cell viability assay

[0599] The purpose of this cellular assay is to determine the effect of the tested compounds on the proliferation of human cancer cell lines NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL-1739) cells, SW620 (ATCC-CCL-227) over a 3-day treatment period by quantifying the amount of NADPH present at the endpoint using the Cell Counting Kit-8.

[0600] Cells were seeded in 96-well assay plates (Corning-3699) at 5,000 cells / well (NCI-H358), 2,000 cells / well (AGS), and 2,000 cells / well (SW620) and incubated overnight. On the day of the assay, diluted compounds were added at a final concentration of 0.5% DMSO. After 72 hours of incubation, one-tenth the volume of Cell Counting Kit 8 (Dnjindo-CK04) was added to each well. After 2 hours of incubation, the signal was read using EnVision (OD450 minus OD650). IC 50 Determined by fitting a 4-parameter sigmoidal concentration response model.

[0601] Table 3. Activity of Examples and Compounds of the Invention in KRAS Cell Viability Assay

[0602]

[0603]

[0604] Example 23

[0605] Interaction assay of KRAS-BRAF and CYPA (500 nM)

[0606] In this example, TR-FRET is also used to measure compound or compound-CYPA-dependent destruction of the KRAS G12C-BRAF complex. The protocol is also used to measure the destruction of KRAS G12D or KRAS G12V binding to BRAF by the compounds of the present invention, respectively. In a 25mM HEPES PH = 7.4 (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, Thermo, 15630080), 0.002% Tween20, 0.1% BSA, 100mM NaCl, 5mM MgCl2, 10μM GMPPNP (guanosine 5'-[β,γ-imino] triphosphate trisodium salt hydrate, Sigma, G0635), unlabeled CYPA, GMPPNP loaded with 6His-KRAS protein and GST-BRAF RBD Mixed in the wells of a 384-well assay plate at final concentrations of 50nM, 6.25nM and 1nM, respectively. The compounds were present in the wells of the plate in a 16-point 3-fold dilution series starting from a final concentration of 10μM and incubated for 3 hours. A mixture of MAb anti-6His-XL665 (Cisbio, 61HISXLB) and Mab anti-GST-TB cryptate (Cisbio, 61GSTTLB) was then added at final concentrations of 6.67nM and 0.21nM, respectively, and the plate was incubated for another 1.5 hours. The TR-FRET signal was read on a PHERstar FSX microplate reader (Ex320 nm, Em 665 / 615nm). Compounds that promote the destruction of the KRAS-BRAF complex were identified as compounds that cause a decrease in the TR-FRET ratio compared to the DMSO control wells. Table 4. Activity of Examples and Compounds of the Present Invention in the Interaction Assay of KRAS-BRAF and CYPA (500nM)

[0607] Examples G12C IC50(μM) G12D IC50(μM) G12V IC50(μM) A555 0.005 0.043 0.023 Example 1 0.046 0.147 0.070 Example 5 0.050 0.089 0.129 Example 6 0.023 0.071 0.061 Example 7 0.095 0.472 0.611 Example 9 0.049 0.225 0.124 Example 10 0.061 0.186 0.149 Example 11 0.024 0.112 0.059 Example 12 0.029 0.140 0.085 Example 13 >10 >10 >10 Example 14 >10 >10 >10 Example 15 0.892 1.648 0.924 Example 16 0.026 0.059 0.097 Example 17 0.002 0.007 0.004

[0608] Example 24

[0609] pERK inhibition assay

[0610] The assay is designed to measure the ability of the tested compounds to inhibit ERK phosphorylation, KRAS G12C in NCI-H358 cells, KRAS G12D in AGS cells, and KRAS G12V downstream signaling in SW620. NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL-1739) cells, and SW620 (ATCC-CCL-227) cells were grown and maintained in RPMI-1640 medium (Thermo Fisher Scientific) containing 10% fetal bovine serum and 1% penicillin / streptomycin. The day before the addition of the compound, the cells were plated at a density of 30,000 cells / well, 20,000 cells / well, and 30,000 cells / well (for NCI-H358, AGS, and SW620) in a tissue culture treated 96-well plate (Corning-3699) and allowed to attach overnight. The diluted compounds were then added at a final concentration of 0.5% DMSO. After 4 hours of incubation, the culture medium was removed, 100 μL 4% formaldehyde was added, and the assay plate was incubated at room temperature for 20 minutes. The plate was then washed once with phosphate buffered saline (PBS) and permeabilized with 100 μL frozen methanol for 10 minutes. Nonspecific antibodies bound to the plate were blocked at room temperature for at least 1 hour using 50 μL 1X BSA blocking buffer (Thermo-37520, diluted 10 times with phosphate buffered saline (PBST)).

[0611] The amount of phosphor-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, with 50 μL aliquoted to each well and incubated overnight at 4 ° C. The cells were washed five times with PBST for 5 minutes. The secondary antibody (HRP-linked anti-rabbit IgG, CST-7074, Cell Signaling Technology) was diluted 1:1000 in blocking buffer, and 50 μL was added to each well, and incubated for 1-2 hours at room temperature. The cells were washed 5 times with PBST for 5 minutes, 100 μL TMBELISA substrate (abcam-ab171523) was added, and gently shaken for 20 minutes. 50 μL stop solution (abcam-ab171529) was added, and then the signal (OD450) was read with EnVision.

[0612] IC 50 Determined by fitting a 4-parameter sigmoid concentration response model.

[0613] Table 5. Activity of Examples and Compounds of the Invention in the KRAS pERK Inhibition Assay

[0614]

[0615]

[0616] Example 25

[0617] Stable KRAS mutant cell lines and cell viability assays.

[0618] The purpose of this study is to use The potency and efficacy of compounds on cell proliferation were determined using the (CTG) luminescent cell viability assay (Promega Corp., Madison, WI). We compared 14 KRAS G12C Variant sequences (V8A, V9Y, S17E, T58I, A59T, S65W, R68S, D69P, M72I, D92R, H95N, Y96D, Q99F, Q99W, Y96H, and F156L) were cloned into Miapaca-2. A total of 14 stable Miapaca-2 mutant cell lines were established by lentiviral infection. For cell viability assays, compounds were administered to cells in a 9-point dose response using a 4-fold dilution series with a maximum dose of 10 μM. KRAS mutant cells were maintained in DMEM + 10% FBS + 2.5% HI horse serum + 1% PS + 1 μg / mL puromycin and seeded in 96-well plates at 800-1,500 cells per well 24 h before adding compounds, and then incubated with compounds for 3 days before measuring viability (CellTiter-Glo, Promega). Assays were performed in biological duplicates. Nonlinear regression curves were fitted using Xfit. IC50 (absolute IC50) is the dose at which the estimated activity relative to untreated wells is 50%. The inhibition rate of the compound was calculated as follows: % inhibition = 100-100 x (luminescence value-HPE) / (ZPE-HPE).

[0619] HPE: Luminescence value from wells with culture medium only

[0620] ZPE: Luminescence value from wells with DMSO

[0621] Table 8 Cell viability in mutant cells with KRAS G12C and other mutations

[0622]

Claims

1. A compound of formula (I), in R 1 for 3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazinyl or (C 1-6 Alkyl) oxoimidazolidinyl; Where R 8 C 1-6 alkyl; R 9 for ((C 1-6 alkyl)2amino)azetidinyl, C 1-6 Alkylpiperazinyl, halogenated azetidinyl, halogenated C 1-6 Alkylamino, halogenated C 1-6 Alkylaminoazetidinyl, halo C 1-6 Alkylpiperazine, hydroxyl (C 1-6 alkyl)piperidinyl or morpholinyl; R 2 C 1-6 alkyl; R 3 is H or halogen; R 4 is H or halogen; R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl; R 6 C 1-6 Alkoxy C 1-6 alkyl; R 7 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl; A 1 is thiazolylidene; A 2 C 1-6 Alkylene; The prerequisite is R 3 and R 4 Not at the same time H; or a pharmaceutically acceptable salt thereof.

2. A compound of formula (Ia), in R 1 for 3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazinyl or (C 1-6 Alkyl) oxoimidazolidinyl; Where R 8 C 1-6 alkyl; R 9 for ((C 1-6 alkyl)2amino)azetidinyl, C 1-6 Alkylpiperazinyl, halogenated azetidinyl, halogenated C 1-6 Alkylamino, halogenated C 1-6 Alkylaminoazetidinyl, halo C 1-6 Alkylpiperazine, hydroxyl (C 1-6 alkyl)piperidinyl or morpholinyl; R 2 C 1-6 alkyl; R 3 is H or halogen; R 4 is H or halogen; R 5 C 1-6 Alkyl or halogenated C 1-6 alkyl; R 6 C 1-6 Alkoxy C 1-6 alkyl; R 7 For morpholinyl, (halogenated C 1-6 alkyl) piperazinyl or C 1-6 Alkylpiperazinyl; A 1 is thiazolylidene; A 2 C 1-6 Alkylene; The prerequisite is R 3 and R 4 Not at the same time H; or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 1 or 2, wherein R 1 for Where R 8 C 1-6 Alkyl; R 9 C 1-6 Alkylpiperazinyl, halogenated C 1-6 Alkylpiperazinyl or morpholinyl.

4. A compound according to any one of claims 1 to 3, wherein R 1 for Where R 8 is methyl; R 9 It is 4-methylpiperazin-1-yl, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or morpholinyl.

5. A compound according to any one of claims 1 to 4, wherein R 1 It is methyl-(4-methylpiperazine-1-carbonyl)amino, methyl-[4-(2,2,2-trifluoroethyl)piperazine-1-carbonyl]amino or methyl(morpholine-4-carbonyl)amino.

6. A compound according to any one of claims 1 to 5, wherein R 2 It is isopropyl.

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

8. A compound according to any one of claims 1 to 7, wherein R 3 For fluorine.

9. A compound according to any one of claims 1 to 8, wherein R 4 is H or fluorine.

10. A compound according to any one of claims 1 to 9, wherein R 4 For H.

11. A compound according to any one of claims 1 to 10, wherein R 5 Halogenated C 1-6 alkyl.

12. A compound according to any one of claims 1 to 11, wherein R 5 It is 2,2,2-trifluoroethyl.

13. A compound according to any one of claims 1 to 12, wherein R 6 It is 1-methoxyethyl.

14. A compound according to any one of claims 1 to 13, wherein R 7 For (halogenated C 1-6 alkyl)piperazinyl.

15. A compound according to any one of claims 1 to 14, wherein R 7 It is 4-(2,2,2-trifluoroethyl)piperazin-1-yl.

16. A compound according to any one of claims 1 to 15, wherein A 1 for Where bond "a" is attached to the indole ring.

17. A compound according to any one of claims 1 to 16, wherein A 2 It is 2,2-propylene.

18. The compound according to claim 1 or 2, wherein R 1 for Where R 8 C 1-6 Alkyl; R 9 C 1-6 Alkylpiperazinyl, halogenated C 1-6 Alkylpiperazinyl or morpholinyl; R 2 C 1-6 alkyl; R 3 is a halogen; R 4 is H; R 5 Halogenated C 1-6 alkyl; R 6 C 1-6 Alkoxy C 1-6 alkyl; R 7 For (halogenated C 1-6 alkyl)piperazinyl; A 1 for wherein bond "a" is attached to the indole ring; A 2 C 1-6 Alkylene; or a pharmaceutically acceptable salt thereof.

19. The compound according to claim 18, wherein R 1 is methyl-(4-methylpiperazine-1-carbonyl)amino, methyl-[4-(2,2,2-trifluoroethyl)piperazine-1-carbonyl]amino or methyl(morpholine-4-carbonyl)amino; R 2 is isopropyl; R 3 For fluorine; R 4 is H; R 5 It is 2,2,2-trifluoroethyl; R 6 is (1S)-1-methoxyethyl; R 7 is 4-(2,2,2-trifluoroethyl)piperazin-1-yl; A 1 for wherein bond "a" is attached to the indole ring; A 2 is 2,2-propylene; or a pharmaceutically acceptable salt thereof.

20. A compound selected from: 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide; 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide; N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide; 3-(Dimethylamino)-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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide; 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]- N-methyl-morpholine-4-carboxamide; N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide; 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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-3,3-difluoro-N-methyl-azetidine-1-carboxamide; (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(3-methyl-2-oxo-imidazolidin-1-yl)butanamide; 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-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide; N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]- N-methyl-morpholine-4-carboxamide; N-[(1S)-1-[[(7S,13S)-24-fluoro-20-(20M)-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide; 3-(Dimethylamino)-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-azetidine-1-carboxamide; (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-[methyl(2,2,2-trifluoroethylcarbamoyl)amino]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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]- N-methyl-3-(trifluoromethyl)azetidine-1-carboxamide; (2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-(3-oxo-5,6,8,8a-tetrahydro-1H-imidazo[5,1-c][1,4]oxazin-2-yl)butanamide; (3R)-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-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-3-hydroxy-N,3-dimethyl-piperidine-1-carboxamide; N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-morpholine-4-carboxamide; N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N,4-dimethyl-piperazine-1-carboxamide; and N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridinyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octacosyl-1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-4-(2,2,2-trifluoroethyl)piperazine-1-carboxamide; or a pharmaceutically acceptable salt thereof.

21. A method for preparing a compound according to any one of claims 1 to 20, comprising the steps of: a) in the presence of a coupling agent and a base, a compound of formula (II), With acid (III), A coupling reaction is carried out between them to form the compound of formula (I); Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , A 1 and A 2 As defined in any one of claims 1 to 19; the coupling reagent is T3P, HATU, PyBOP or EDCI / HOBt; the base is TEA, DIEPA or DMAP.

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

23. A pharmaceutical composition comprising a compound according to any one of claims 1 to 20 and a pharmaceutically acceptable excipient.

24. Use of a compound according to any one of claims 1 to 20 for treating a KRAS G12C protein-related disease.

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

26. Use of a compound according to any one of claims 1 to 20 for inhibiting the interaction of RAS with downstream effectors, wherein the downstream effectors are RAF and PI3K.

27. Use of a compound according to any one of claims 1 to 20 for inhibiting propagated oncogenic MAPK and PI3K signalling.

28. Use of a compound according to any one of claims 1 to 20 for treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma, ovarian cancer and endometrial cancer.

29. Use of a compound according to any one of claims 1 to 20 for treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

30. A compound or pharmaceutically acceptable salt according to any one of claims 1 to 20 for use in treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

31. Use of a compound according to any one of claims 1 to 20 for treating or preventing a KRAS mutation 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.

32. Use of a compound according to any one of claims 1 to 20 for the preparation of a medicament for treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer.

33. Use of a compound according to any one of claims 1 to 18 for the preparation of a medicament for treating or preventing a KRAS mutation-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.

34. A method for treating or preventing a KRAS mutation driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer, the method comprising administering a therapeutically effective amount of a compound as defined in any one of claims 1 to 20.

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

36. A compound or pharmaceutically acceptable salt according to any one of claims 1 to 20, prepared according to the process of claim 21.

37. The invention as herein described.

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