HK40103332A - Macrocyclic compounds for the treatment of cancer - Google Patents

Description

Macrocyclic compounds used to treat cancer

This invention relates to organic compounds that can be used for treatment and/or prevention in mammals, and particularly to inhibitory effects on KRAS G12C that can be used to treat cancer.

Technical Field

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

All KRAS proteins belong to the family of small GTPases that hydrolyze GTP to GDP. Structurally, KRAS is divided into an effector-binding leaflet, followed by an allosteric leaflet and a C-terminal region responsible for membrane anchoring. The effector leaflet includes the P-loop, switch I, and switch II regions. The switch I/II loops play a crucial role in downstream KRAS signaling by mediating protein-protein interactions with effector proteins, including RAF in the mitogen-activated protein kinase (MAPK) pathway or PI3K in the phosphatidylinositol 3-kinase (P13K)/protein kinase B (AKT) pathway.

KRAS proteins switch between inactive and active forms via binding to GTP and GDP, respectively. Under physiological conditions, the transition between these two states is regulated by guanine nucleotide exchange factors (GEFs), such as SOS1 (Seven Sons homolog 1) or GTPase activators (GAPs) involved in catalyzing the exchange of GDP for GTP, thereby enhancing intrinsic GTPase activity or accelerating RAS-mediated GTP hydrolysis. In response to extracellular stimuli, inactive RAS-GDP is converted to active RAS-GTP, which directly binds to the RAF RAS-binding domain (RAF ^RBD ), thereby recruiting the RAF kinase family from the cytoplasm to the cell membrane, where they dimerize and become active. Activated RAFs then undergo a series of phosphorylation reactions on their downstream mitogen-activated protein kinases (MEK) and extracellular signal-regulated kinases (ERK), and propagate growth signals. Among the RAF protein kinase family (three known isoforms ARAF, BRAF, and CRAF/RAF1), BRAF mutations are most frequent and it remains the most potent activator of MEK. Although individual RAS and RAF family members exhibit different binding preferences, all RAFs possess a conserved RBD for forward transmission of MAPK signaling, frequently used to characterize KRAS inhibition (e.g., the KRAS-BRAF ^RBD in this paper). For KRAS, mutations at positions 12, 13, 61, and 146 lead to conversion to the active KRAS form by weakening nucleotide hydrolysis or activating nucleotide exchange, thereby resulting in overactivation of the tumorigenetic MAPK pathway.

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

Summary of the Invention

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

in

^R1 is 6-oxo-2,7-diazaspiro[4.5]decane-7-yl or substituted with a (dihalo _-C1-6 alkyl) carbonyl group;

Wherein ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group.

Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl,

The piperidinyl group is substituted once or twice with a substituent independently selected from halogen, (dihalo _-C1-6 alkyl) carbonyl, and _C2-6 ynyl, or

Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H, morpholino, (halogenated _C1-6 alkyl)piperazinyl or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

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

Compounds of formula (I) or (Ia) exhibit good KRAS inhibitory activity against G12C, G12D, and G12V. In another embodiment, the compounds of the present invention exhibit excellent cancer cell inhibitory activity. Furthermore, compounds of formula (I) or (Ia) also exhibit good or improved cytotoxicity and solubility characteristics. Moreover, compared to reference compounds, the compounds of the present invention address the GSH toxicity problem (see Example 27).

Attached Figure Description

Figure 1. X-ray crystallographic analysis of interstitial body A

Detailed Implementation

definition

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

The term "( _C1-6 )alkylene" refers to a straight-chain or branched saturated divalent hydrocarbon group with 1 to 6 carbon atoms, or a divalent branched saturated divalent hydrocarbon group with 3 to 6 carbon atoms. Examples of ( _{C1-6 )} alkylene groups include methylene, ethylene, propylene, 2-methylpropylene, butylene, 2-ethylbutylene, pentylene, and hexylene.

The term "C _2-6 alkenyl" refers to a monovalent straight-chain or branched hydrocarbon group with 2 to 6 carbon atoms and at least one double bond. In certain embodiments, the alkenyl group has 2 to 4 carbon atoms and at least one double bond. Examples of C _2-6 alkenyl groups include ethenyl, propenyl, prop-2-enyl, isopropenyl, n-butenyl, and isobutenyl.

The term "C _2-6 ynyl" refers to a monovalent, straight-chain or branched saturated hydrocarbon group having 2 to 6 carbon atoms, including one, two, or three triple bonds. In certain embodiments, the ynyl group has 2 to 4 carbon atoms, including one or two triple bonds. Examples of _{C 2-6} ynyl groups include ethynyl, propynyl, prop-2-ynyl, isopropynyl, and n-butynyl.

The term "C _3-8 dienyl" refers to a hydrocarbon with 3 to 8 carbon atoms containing two double bonds between the carbon atoms. These two double bonds can be located at different positions in the molecule, attached to the same carbon atom (accumulated, C=C=C), or separated from each other by a single σ bond (conjugated, C=CC=C). Examples of "C _3-8 dienyl" include butadiene.

The term "cycloalkyl" refers to a monovalent saturated monocyclic or bicyclic hydrocarbon group with 3 to 10 ring carbon atoms. In specific embodiments, cycloalkyl refers to a monovalent saturated monocyclic hydrocarbon group with 3 to 8 ring carbon atoms. Bicyclic means consisting of two saturated carbon rings having one or two common carbon atoms. Examples of monocyclic cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl. Examples of bicyclic cycloalkyl are bicyclic [1.1.0]butyl, bicyclic [2.2.1]heptyl, or bicyclic [2.2.2]octyl.

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

The term "dihalo _-C1-6 alkyl" refers to a _C1-6 alkyl group in which two hydrogen atoms of the _C1-6 alkyl group have been replaced by the same or different halogen atoms. Examples of dihalo _-C1-6 alkyl groups include difluoro- or chloro(fluoro)-methyl, -ethyl, or -propyl, such as difluoropropyl, difluoromethyl, difluoroethyl, or chloro(fluoro)methyl.

The term "halogenated _C1-6 alkyl" refers to a _C1-6 alkyl group in which at least one hydrogen atom of the _C1-6 alkyl group has been substituted with the same or a different halogen atom. Examples of halogenated _C1-6 alkyl groups include fluorine, difluoro- or chloro(fluoro)-methyl, -ethyl or -propyl, such as fluoromethyl, difluoropropyl, difluoromethyl, difluoroethyl, chloro(fluoro)methyl, trifluoroethyl or trifluoromethyl.

The term "phenylene" refers to a divalent phenyl group.

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

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

The term "thiol group" refers to the -S- group.

The term "heterocyclic group" refers to a monocyclic or bicyclic system with 3 to 9 ring atoms that is monovalently saturated or partially unsaturated, containing 1, 2, or 3 heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. In specific embodiments, a heterocyclic group is a monovalently saturated monocyclic system with 4 to 7 ring atoms that contains 1, 2, or 3 cyclic heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. Examples of monocyclic saturated heterocyclic groups are aziridinyl, ethylene oxide, aziridine, oxadiazinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydro-thiophenyl, pyrazolyl, imidazoyl, oxazolyl, isoxazolyl, thiazoyl, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, aziridine-heptyl, diaziridine-heptyl, homopiperazinyl, or oxadiazinyl-heptyl. Examples of bicyclic saturated heterocyclic groups are 8-aza-bicyclo[3.2.1]octyl, quininecyclo, 8-oxa-3-aza-bicyclo[3.2.1]octyl, 9-aza-bicyclo[3.3.1]nonyl, 3-oxa-9-aza-bicyclo[3.3.1]nonyl, or 3-thia-9-aza-bicyclo[3.3.1]nonyl. Examples of partially unsaturated heterocyclic groups are dihydrofuranyl, imidazolinyl, dihydrooxazolyl, tetrahydropyridyl, or dihydropyranyl. The term "sub-heterocyclic group" refers to a divalent heterocyclic group.

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

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

"Pharmaceutical acid addition salts" refer to pharmaceutical salts formed with inorganic and organic acids. The inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, and phosphoric acid. The organic acids can be selected from aliphatic, alicyclic, aromatic, arylaliphatic, heterocyclic, formic, and sulfonic acid organic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, dihydroxynaphthalic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid.

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

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

The term "therapeutic effective amount" refers to the amount of a compound or molecule of the present invention, when administered to a subject, (i) to treat or prevent a particular disease, condition, or disorder; (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder; or (iii) to prevent or delay the onset of one or more symptoms of a particular disease, condition, or disorder described herein. Therapeutic effective amount depends on the compound, the disease state being treated, the severity of the disease being treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending physician or veterinarian, and other factors.

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

KRAS inhibitors

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

in

^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with (dihalo _-C1-6 alkyl) carbonyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with (dihalo _-C1-6 alkyl) carbonyl;

Wherein ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group.

Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl,

The piperidinyl group is substituted once or twice with a substituent independently selected from halogen, (dihalo _-C1-6 alkyl) carbonyl, and _C2-6 ynyl, or

Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups;

R ⁷ is a _C1-6 alkyl group;

_R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H, morpholino, (halogenated _C1-6 alkyl)piperazinyl or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

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

in

^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with (dihalo _-C1-6 alkyl) carbonyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with (dihalo _-C1-6 alkyl) carbonyl;

Wherein ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group.

Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl,

The piperidinyl group is substituted once or twice with a substituent independently selected from halogen, (dihalo _-C1-6 alkyl) carbonyl, and _C2-6 ynyl, or

Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H, morpholino, (halogenated _C1-6 alkyl)piperazinyl or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the present invention is (iii) a compound of formula (I) or (Ia) according to (i) or (ii), or a pharmaceutical salt thereof, wherein ^R1 is 1-oxo- _2,7 -diazaspiro[4.4]nonane-2-yl substituted with a (dihalo- _C1-6 alkyl) carbonyl group or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with a (dihalo-C1-6 alkyl) carbonyl group; wherein R6 is a pyrrolidinyl substituted with a (dihalo _-C1-6 alkyl) carbonyl group; and ^R7 is a _C1-6 alkyl group.

A further embodiment of the invention is (iv) a compound of formula (I) or (Ia) according to any one of (i) to (iii), or a pharmaceutical salt thereof, wherein ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with chloro(fluoro)acetyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with chloro(fluoro)acetyl; wherein R6 is pyrroleyl substituted with chloro(fluoro)acetyl; and R7 is methyl.

A further embodiment of the invention is (v) a compound of formula (I) or (Ia) according to any one of (i) to (iv), wherein ^R1 is

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 pharmaceutical salt thereof, wherein ^R2 is isopropyl.

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 pharmaceutical salt thereof, wherein ^R3 is ethyl or trifluoroethyl.

A further embodiment of the invention is (viii) a compound of formula (I) or (Ia) according to any one of (i) to (vii), or a pharmaceutical salt thereof, wherein ^R4 is methoxyethyl.

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 pharmaceutical salt thereof, wherein ^R4 is

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 pharmaceutical salt thereof, wherein R5 is morpholino or methylpiperazinyl.

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 pharmaceutical salt thereof, wherein ^A1 is a compound in which bond “a” is attached to an indole ring.

A further embodiment of the present invention is (xii) a compound of formula (I) or (Ia) according to any one of (i) to (xi), or a pharmaceutical salt thereof, wherein A2 is a dimethylmethylene.

A further embodiment of the invention is (xiii) a compound of formula (I) or (Ia) according to any one of (i) to (xii), wherein

^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with (dihalo _-C1-6 alkyl) carbonyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with (dihalo _-C1-6 alkyl) carbonyl;

Wherein ^R6 is a pyrrolidinyl group substituted with a (dihalo _-C1-6 alkyl) carbonyl group; ^R7 is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

R ⁵ is morpholino or _C1-6 alkylpiperazino;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the invention is (xiv) a compound of formula (I) or (Ia) according to any one of (i) to (xiii), wherein

^R1 is

^R2 is isopropyl;

^R3 is ethyl or trifluoroethyl;

^R4 is

R5 is either morpholino or methylpiperazino;

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

^A2 is a dimethylmethylene group;

A ³ is O;

Or its medicinal salt.

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

in

^R1 is 6-oxo-2,7-diazaspiro[4.5]decane-7-yl or substituted with a (dihalo _-C1-6 alkyl) carbonyl group;

Wherein ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group.

Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl,

The piperidinyl group is substituted once or twice with a substituent independently selected from halogen, (dihalo _-C1-6 alkyl) carbonyl, and _C2-6 ynyl, or

Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H, morpholino, (halogenated _C1-6 alkyl)piperazinyl or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

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

in

^R1 is 6-oxo-2,7-diazaspiro[4.5]decane-7-yl or substituted with a (dihalo _-C1-6 alkyl) carbonyl group;

R6 is a _C2-6 alkynyl-substituted nitrogen-containing heterocyclic butyl group.

Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6-thionyl] _C1-6 alkyl carbonyl,

The piperidinyl group is substituted once or twice with a substituent independently selected from halogen, (dihalo _-C1-6 alkyl) carbonyl, and _C2-6 ynyl, or

Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H, morpholino, (halogenated _C1-6 alkyl)piperazinyl or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the present invention is (iii′) a compound of formula (I) or (Ia) according to (i′) or (ii′), or a pharmaceutical salt thereof, wherein ^R1 is or is 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with a (dihalo _-C1-6 alkyl) carbonyl group; wherein R6 is a pyrrolidinyl substituted with a (dihalo _-C1-6 alkyl) carbonyl group; and R7 is a _C1-6 alkyl group.

A further embodiment of the present invention is (iv′) a compound of formula (I) or (Ia) according to any one of (i′) to (iii′), or a pharmaceutical salt thereof, wherein ^R1 is or is 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with chloro(fluoro)acetyl; wherein R6 is pyrrolidinyl substituted with chloro(fluoro)acetyl; and ^R7 is methyl.

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 R1 is methyl and R6 is methyl.

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 pharmaceutical salt thereof, wherein ^R2 is isopropyl.

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 pharmaceutical salt thereof, wherein ^R3 is ethyl or trifluoroethyl.

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 pharmaceutical salt thereof, wherein ^R4 is methoxyethyl.

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 pharmaceutical salt thereof, wherein ^R4 is

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 pharmaceutical salt thereof, wherein R5 is morpholino or methylpiperazinyl.

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 pharmaceutical salt thereof, wherein A1 is a compound in which bond “a” is attached to an indole ring.

A further embodiment of the present invention is (xii′) a compound of formula (I) or (Ia) according to any one of (i′) to (xi′), or a pharmaceutical salt thereof, wherein A2 is a dimethylmethylene.

A further embodiment of the present invention is (xiii′) a compound of formula (I) or (Ia) according to any one of (i′) to (xii′), wherein

^R1 is 6-oxo-2,7-diazaspiro[4.5]decane-7-yl or substituted with a (dihalo _-C1-6 alkyl) carbonyl group;

Wherein ^R6 is a pyrrolidinyl group substituted with a (dihalo _-C1-6 alkyl) carbonyl group; ^R7 is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

R ⁵ is morpholino or _C1-6 alkylpiperazino;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the present invention is (xiv′) a compound of formula (I) or (Ia) according to any one of (i′) to (xiii′), wherein

^R1 is

Where ^R6 is methyl and ^R7 is methyl;

^R2 is isopropyl;

^R3 is ethyl or trifluoroethyl;

^R4 is

R ⁵ is morpholino or methylpiperazino;

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

^A2 is a dimethylmethylene group;

A ³ is O;

Or its medicinal salt.

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

in

^R1 is

Wherein ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group.

Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl,

The piperidinyl group is substituted once or twice with a substituent independently selected from halogens and _C2-6 ynyl groups, or

Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl group;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

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

in

^R1 is

Wherein ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group.

Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl,

The piperidinyl group is substituted once or twice with a substituent independently selected from halogens and _C2-6 ynyl groups, or

Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl group;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the present invention is (iii″) a compound of formula (I) or (Ia) according to (i″) or (ii″), or a pharmaceutical salt thereof, wherein ^R1 is a pyrrolidinyl substituted with a (dihalo _-C1-6 alkyl) carbonyl group; and ^R7 is a _C1-6 alkyl group.

A further embodiment of the invention is (iv″) a compound of formula (I) or (Ia) according to any one of (i″) to (iii″), or a pharmaceutical salt thereof, wherein ^R1 is a ^pyrrolidinyl group substituted with chloro(fluoro)acetyl; and ^R7 is methyl.

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

^R1 is where ^R6 is where ^R7 is a methyl group.

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 pharmaceutical salt thereof, wherein ^R2 is isopropyl.

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 pharmaceutical salt thereof, wherein ^R3 is ethyl.

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 pharmaceutical salt thereof, wherein ^R4 is methoxyethyl.

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 pharmaceutical salt thereof, wherein R4 is

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 pharmaceutical salt thereof, wherein ^R5 is H or methylpiperazinyl.

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 pharmaceutical salt thereof, wherein ^A1 is

A further embodiment of the present invention is (xii″) a compound of formula (I) or (Ia) according to any one of (i″) to (xi″), or a pharmaceutical salt thereof, wherein A2 is a dimethylmethylene.

A further embodiment of the present invention is (xiii″′) a compound of formula (I) or (Ia) according to any one of (i″″) to (xii″″), wherein

^R1 is

Wherein ^R6 is a pyrrolidinyl group substituted with a (dihalo _-C1-6 alkyl) carbonyl group; ^R7 is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl group;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R5 is H or _C1-6 alkylpiperazinyl;

^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the present invention is (xiv″) a compound of formula (I) or (Ia) according to any one of (i″) to (xiii″), wherein

^R1 is

Where ^R6 is methyl and ^R7 is methyl;

^R2 is isopropyl;

^R3 is an ethyl group;

^R4 is

^R5 is H or methylpiperazinyl;

A ¹ is

^A2 is a dimethylmethylene group;

A ³ is O;

Or its medicinal salt.

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

in

^R1 is

^R6 is a pyrrolidinyl group substituted with ( _C1-6 alkyl) _2- oxooxetanebutylcarbonyl, ( _C1-6 alkylcarbonyl)carbonyl, ( _dihaloC1-6 alkyl)carbonyl, (oxooxetanebutylamino)carbonyl, _C2-6 ynyl, _C3-8 dienylcarbonyl, _cyanoC1-6 alkyl, cycloalkylcarbonyl, oxonitrilecyclobutylcarbonyl, pyridylC2-6 ynylcarbonyl, or _{triazolylC2-6} alkenylcarbonyl; or a cycloalkyl group substituted with formyl, _C2-6 ynyl, _C1-6 alkylsulfonylC2-6 alkenyl, ( _C1-6 alkyl) _{2 phosphorylC2-6} alkenyl, or [( _C1-6 alkyl) ₂ ⁽ oxo _{) -λ6} -thionyl] _C1-6 alkylcarbonyl.

R ⁷ is a _C1-6 alkyl group;

^R8 is ((dihalo _-C1-6 alkyl)carbonyl)azacyclobutane;

^A4 is a _C1-6 alkylene group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl group;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

R ⁵ is H, (C _1-6 alkyl) _2- phosphoryl, C _1-6 alkylpiperazinyl, or C _1-6 alkylsulfinyl;

^A1 is an thiazolyl or phenylene group substituted with ^R9 ; wherein ^R9 is H, hydroxyl, ( _C1-6 alkyl) _2- phosphoryl or _C1-6 alkylsulfinyl.

^A2 is a _C1-6 alkylene group;

^A3 is O or S;

Or its medicinal salt.

A further embodiment of the present invention is (ii″′) a compound of formula (I) according to (1″′), wherein

^R1 is

^R6 is a pyrrolidinyl substituted with ( _C1-6 alkyl carbonyl) carbonyl, (dihalo _C1-6 alkyl) carbonyl, _C2-6 ynyl or cyano _C1-6 alkyl; or a cycloalkyl substituted with formyl, _C2-6 ynyl or [( _C1-6 alkyl) ₂ (oxo) ^-λ6 -thionyl] _C1-6 alkyl carbonyl;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl group;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

R ⁵ is H;

^A1 is a phenylene substituted with ^R9 ; where ^R9 is a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the present invention is (iii″′) a compound of formula (I) according to (i″′) or (ii″′), or a pharmaceutical salt thereof, wherein

^R1 is

^R6 is a pyrrolidinyl substituted with (dihalo _-C1-6 alkyl)carbonyl, or a cycloalkyl substituted with formyl, _C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo) ^-λ6 -thionyl] _C1-6 alkylcarbonyl;

^R7 is a _C1-6 alkyl group.

A further embodiment of the present invention is (iv″′) a compound of formula (I) according to any one of (i″′) to (iii″′), or a pharmaceutical salt thereof, wherein

^R1 is

^R6 is a pyrrole alkyl group substituted with chloro(fluoro)acetyl, or a cycloalkyl group substituted with formyl, ethynyl or [dimethyl(oxo) ^-λ6 -thionyl]acetyl;

^R7 is a methyl group.

A further embodiment of the present invention is (v″′) a compound of formula (I) according to any one of (i″′) to (iv″′), wherein

^R1 is where ^R6 is where ^R7 is a methyl group.

A further embodiment of the present invention is (vi″′) a compound of formula (I) according to any one of (i″′) to (v″′), or a pharmaceutical salt thereof, wherein ^R2 is isopropyl.

A further embodiment of the present invention is (vii″′) a compound of formula (I) according to any one of (i″′) to (vi″′), or a pharmaceutical salt thereof, wherein ^R3 is ethyl.

A further embodiment of the present invention is (viii″′) a compound of formula (I) according to any one of (i″′) to (vii″′), or a pharmaceutical salt thereof, wherein ^R4 is methoxyethyl.

A further embodiment of the present invention is (ix″′) a compound of formula (I) according to any one of (i″′) to (viii″′), or a pharmaceutical salt thereof, wherein ^R5 is H.

A further embodiment of the present invention is (x″′) a compound of formula (I) according to any one of (i″′) to (ix″′), or a pharmaceutical salt thereof, wherein A ¹ is a hydroxyl ^group .

A further embodiment of the present invention is (xi″′′) a compound of formula (I) according to any one of (i″′) to (x″′), or a pharmaceutical salt thereof, wherein ^A2 is

A further embodiment of the present invention is (xii″′) a compound of formula (I) according to any one of (i″′) to (xi″′), wherein

^R1 is

^R6 is a pyrrolidinyl substituted with (dihalo _-C1-6 alkyl)carbonyl, or a cycloalkyl substituted with formyl, _C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo) ^-λ6 -thionyl] _C1-6 alkylcarbonyl;

R ⁷ is a _C1-6 alkyl group;

^R2 is a _C1-6 alkyl group;

^R3 is a _C1-6 alkyl group;

^R4 is a _C1-6 alkoxy- _C1-6 alkyl group;

R ⁵ is H;

^A1 is a phenylene substituted with ^R9 ; where ^R9 is a hydroxyl group;

^A2 is a _C1-6 alkylene group;

A ³ is O;

Or its medicinal salt.

A further embodiment of the present invention is (xii″′) a compound of formula (I) according to any one of (i″′) to (xi″′), wherein

^R1 is

Where ^R6 is

^R7 is a methyl group;

^R2 is isopropyl;

^R3 is an ethyl group;

^R4 is methoxyethyl;

R ⁵ is H;

^A1 represents R9, where ^R9 is a hydroxyl group;

^A2 is

A ³ is O;

Or its medicinal salt.

Another embodiment of the present invention is a compound of formula (I), which is selected from the following compounds:

(3S)-1-[(2S)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-formyl-N-methyl-cyclobutaneformamide;

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-ethynyl-N-methyl-cyclobutanecarboxamide;

(3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-(2-oxopropionyl)pyrrolidine-3-carboxamide;

(3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-prop-2-ynylpyrrolidine-3-carboxamide;

(3S)-1-(cyanomethyl)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

3-[2-[dimethyl(oxo) ^-λ6 -thionyl]acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-cyclobutanecarboxamide;

(3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(3S)-1-(bicyclo[1.1.0]butane-1-carbonyl)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxamide;

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-ethynyl-N-methyl-azacyclobutane-1-carboxamide;

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-4-ethynyl-N-methyl-piperidine-1-carboxamide;

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-4-ethynyl-4-fluoro-N-methyl-piperidine-1-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoroacetyl]-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl [8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butanamide;]

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl [8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butanamide;]

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; and

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

Or its medicinal salt.

Another embodiment of the present invention is a compound RM461-A having the structure of (xvi), or a pharmaceutical salt thereof.

Another embodiment of the present invention relates to (xvii) a method for preparing a compound according to any one of (i) to (xv), (i′) to (xiv′), (i″) to (xiii″) or (1″′) to (xii′″), the method comprising any one of the following steps:

a) Using a coupling agent in the presence of a base, compound of formula (IX) is made into a compound.

With acid (X),

Coupled reactions occur between them;

b) Using a coupling agent in the presence of a base, compound (XIII) is converted into a compound.

It undergoes a coupling reaction with the acid (XIV);

c) Using a coupling agent in the presence of a base to convert compound (XVIII)

It undergoes a coupling reaction with the acid (XIX);

Wherein Q is a heterocyclic group; T is ( _C1-6 alkyl) _2- oxooxetane, _C1-6 alkyl carbonyl, dihaloC1-6 alkyl, oxooxetane _amino , _C2-6 ynyl, _C3-8 dienyl, _cyanoC1-6 alkyl, cycloalkyl, morpholinoC2-6 ynyl, oxoazacyclic butane, _pyridylC2-6 ynyl, or triazolylC2-6 alkenyl; ^R1 , ^R2 , R3, ^R4 , ^R5 , ^R6 , ^R7 , ^A1 , ^A2 , ^{and A3} are as defined in any one of claims 1 to 14; the coupling agent is _T3P , _HATU , PyBOP, or EDCI/ _HOBt ; the base is TEA, DIEPA, or DMAP.

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

Another embodiment of the present invention is (xix) a pharmaceutical composition comprising a compound according to any one of (i) to (xvi) and a therapeutically inert carrier.

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

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

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

Another embodiment of the invention is (xxiii) the use of the compound according to any one of (i) to (xvi) for inhibiting the propagation of carcinogenic MAPK and PI3K signaling.

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

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

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

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

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

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

Pharmaceutical composition and administration

Another embodiment provides pharmaceutical compositions or medicaments comprising the compounds of the present invention and a therapeutically inert carrier, diluent, or excipient, and methods for preparing such compositions and medicaments using the compounds of the present invention. In one example, the compound of formula (I) can be formulated into a galenical form by mixing it with a physiologically acceptable carrier (i.e., a carrier that is non-toxic to the recipient at the dose and concentration used) at ambient temperature, at a suitable pH, and with the desired purity. The pH of the formulation depends primarily on the specific use and concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, the compound of formula (I) is formulated in an acetate buffer at pH 5. In another embodiment, the compound of formula (I) is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

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

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

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

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

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

Examples of suitable oral dosage forms are tablets containing about 1 to 1000 mg of the compound of the present invention combined with about 1 to 1000 mg of anhydrous lactose, about 1 to 1000 mg of croscarmellose sodium, about 1 to 1000 mg of polyvinylpyrrolidone (PVP) K30 and about 1 to 1000 mg of magnesium stearate. The powdered ingredients are first mixed together, and then mixed with a PVP solution. The resulting composition can be dried, granulated, mixed with magnesium stearate, and compressed into tablet form using conventional equipment. Examples of aerosol formulations can be prepared by dissolving the compound of the present invention (e.g., 5 mg to 400 mg) in a suitable buffer solution (e.g., phosphate buffer), with the addition of a permeation enhancer (e.g., a salt such as sodium chloride) if desired. The solution can be filtered, for example, using a 0.2-micron filter to remove impurities and contaminants.

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

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

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

Example A

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

Example B

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

Indications and treatment methods

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

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

synthesis

The compounds of this invention can be prepared by any conventional method. Suitable methods for synthesizing these compounds and their starting materials are provided in the following schemes and examples. Unless otherwise stated, all substituents, particularly R1 to R5 and A1 to ^A3, are as defined above. Furthermore, unless explicitly stated otherwise, all reactions, reaction conditions, abbreviations, and symbols have meanings well known to those skilled in the art of organic chemistry.

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

Option 1

Where X is a halogen; PG is a protecting group, such as Boc and Cbz; and Q is a heterocyclic group.

The coupling of compound (II) with compound (III) can be achieved under standard Suzuki coupling conditions to provide compound (IV). Compound (VII) can be obtained by coupling acid (V) with compound (VI) in the presence of bases such as TEA, DIEPA, and DMAP using coupling agents such as _T3P , HATU, PyBOP, and EDCI/HOBt. The coupling of compound (IV) with compound (VII) can be achieved under Suzuki coupling conditions to provide compound (VIII). Deprotection of compound (VIII) can be achieved in the presence of acid such as TFA, or under hydrogenation conditions using catalysts such as Pd/C and Pd(OH) ₂ /C to provide compound (IX). Compound (I) can be obtained by coupling acid (X) with compound (IX) in the presence of bases such as TEA, DIEPA, and DMAP using coupling agents such as _T3P , HATU, PyBOP, and EDCI/HOBt.

Option 2

Where X is a halogen; PG is a protecting group, such as Boc and Cbz; and Q is a heterocyclic group.

Alternatively, compound (XII) can be obtained by coupling acid (XI) with compound (IX) in the presence of a base such as TEA, DIEPA, or DMAP using a coupling agent such as _T3P , HATU, PyBOP, or EDCI/HOBt. Deprotection of compound (XII) can be achieved in the presence of an acid such as TFA, or under hydrogenation conditions using catalysts such as Pd/C and Pd(OH)2/C, to provide compound (XIII). Compound (XV) can be obtained by coupling acid (XIV) with compound (XIII) in the presence of a base such as TEA, DIEPA, or DMAP using a coupling agent such as _T3P , HATU, PyBOP, or EDCI/HOBt.

Option 3

Where X is a halogen; PG is a protecting group, such as Boc and Cbz; Q is a heterocyclic group; T is (C _1-6 alkyl) ₂ -oxooxetane, C _1-6 alkyl carbonyl, dihaloC _1-6 alkyl, oxooxetane amino, C _2-6 ynyl, C _3-8 dienyl, cyanoC _{1-6 alkyl, cycloalkyl, oxonitrile, pyridylC 2-6 ynyl} or triazolylC _2-6 alkenyl.

Alternatively, compounds of formula (XVII) can be obtained by coupling an acid (XVI) with a compound of formula (XIII) in the presence of a base such as TEA, DIEPA, or DMAP using a coupling agent such as _T3P , HATU, PyBOP, or EDCI/HOBt. Deprotection of compounds of formula (XVII) can be achieved in the presence of an acid such as TFA, or under hydrogenation conditions using catalysts such as Pd/C and Pd(OH)2/C, to obtain compounds of formula (XVIII). Compounds of formula (XX) can be obtained by coupling an acid (XIX) with a compound of formula (XVIII) in the presence of a base such as TEA, DIEPA, or DMAP using a coupling agent such as _T3P , HATU, PyBOP, or EDCI/HOBt.

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

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

a) Using a coupling agent in the presence of a base, compound of formula (IX) is made into a compound.

With acid (X),

Coupled reactions occur between them;

b) Using a coupling agent in the presence of a base, compound (XIII) is converted into a compound.

It undergoes a coupling reaction with the acid (XIV);

c) Using a coupling agent in the presence of a base to convert compound (XVIII)

It undergoes a coupling reaction with the acid (XIX);

in

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

A compound of formula (I), which is manufactured according to the above method, is also the object of this invention.

Example

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

abbreviation

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

The abbreviations used in this article are as follows:

ACN Acetonitrile

aq. Water-based

CDCl ₃ : Deuterated chloroform

_CD3 OD: Deuterated methanol

DIEPA: N,N-Diethylpropylamine

DMAP: 4-Dimethylaminopyridine

DMF: Dimethylformamide

DMSO: Dimethyl sulfoxide

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

EtOAc or EA: Ethyl acetate

FRET (Fluorescence Resonance Energy Transfer)

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

hr: hours

HPLC: High Performance Liquid Chromatography

HOBt: N-hydroxybenzotriazole

MS (ESI): Mass spectrometry (electrospray ionization)

min

NMR: Nuclear Magnetic Resonance

obsd. Observation value

Preparative HPLC: Preparative high-performance liquid chromatography

PyBOP: Benzotriazole-1-yloxytripyrrolidinephosphine hexafluorophosphate

RT or rt: Room temperature

sat. saturated

SFC (Supercritical Fluid Chromatography)

TEA: Triethylamine

TFA: Trifluoroacetic acid

THF: Tetrahydrofuran

TEA: Triethylamine

T ₃ P: Propylphosphonic anhydride

General experimental conditions

Purify intermediates and final compounds using rapid chromatography with one of the following instruments: i) Biotage SP1 system and Quad 12/25 Cartridge module; ii) ISCO combi-flash chromatograph. Silica gel brand and pore size: i) KP-SIL, particle size: 40-60 μm; ii) CAS Registry Number: 63231-67-4, particle size: 47-60 μm silica gel; iii) ZCX from Qingdao Ocean Chemical Co., Ltd., pore size: 200-300 or 300-400.

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

For chiral separation of SFC, intermediates were separated 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 the Mettler Toledo Multigram III SFC system, Waters 80Q preparative SFC, or Thar 80 preparative SFC. The solvent system was _CO2 and IPA (0.5% TEA in IPA solution) or _CO2 and MeOH (0.1% _NH3 · _H2O in MeOH solution), with a back pressure of 100 bar and UV detection at 254 or 220 nm.

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

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

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

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

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

Neutral conditions: A: _H₂O ; B: Acetonitrile.

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

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

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

Preparation Examples

Preparation of intermediates

Intermediate A

3-[5-bromo-1-ethyl-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol

Title intermediate A was prepared according to the following scheme:

Preparation of 2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)pyridine (compound A12)

To a mixture of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (compound A11, 40.0 g, 185.1 mmol) and bis(pinacol)diboron (56.4 g, 222.1 mmol) in 1,4-dioxane (500 mL), KOAc (23.1 mL, 370.2 mmol) and [1,1′-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (6.7 g, 9.2 mmol) were added. The mixture was stirred at 100 °C for 5 hours under _N2 protection. TLC (ethyl acetate) showed that the starting material was consumed. 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 2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)pyridine (compound A12, 55 g), a dark brown oil. MS: calculated value 264 (MH ⁺ ); measured value 264.1 (MH ⁺ ).

Step 1: Preparation of methyl 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionate (compound A2)

At 0 °C, tert-butylchlorodiphenylsilane (256.5 mL, 998.7 mmol) was added to a solution of methyl 3-hydroxy-2,2-dimethylpropionate (compound A1, 110.0 g, 832.3 mmol) and imidazole (169.9 g, 2497 mmol) in THF (1500 mL). The mixture was stirred at 0 °C for 2 hours. The mixture was diluted with petroleum ether (1000 mL) and filtered. The filter cake was washed twice with petroleum ether (150 mL). The combined filtrates were concentrated under vacuum to obtain a residue. The residue was purified by silica gel column chromatography (EA/PE: 0-50%) to give methyl 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionate (compound A2, 220 g) as a colorless oil. ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.68-7.63 (m, 4H), 7.44-7.36 (m, 6H), 3.69 (s, 3H), 3.65 (s, 2H), 1.21 (s, 6H), 1.04 (s, 9H).

Step 2: Preparation of 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionic acid (compound A3)

A solution of potassium hydroxide (43.2 g, 770.2 mmol) in ethanol (500 mL) was added to a solution of methyl 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionate (compound A2, 110.0 g, 296.8 mmol) in ethanol (1200 mL). The mixture was stirred at 90 °C for 5 hours. The mixture was concentrated under vacuum to remove EtOH and diluted with ice water (1000 mL). The mixture was acidified with 1 M HCl aqueous solution until pH 3. The aqueous phase was extracted twice with EtOAc (600 mL). The combined organic layers were washed with brine (400 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum. The residue was ground with petroleum ether (300 mL) to give 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionate (compound A3, 80 g) as a white solid. ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.70-7.65 (m, 4H), 7.47-7.38 (m, 6H), 3.67 (s, 2H), 1.25 (s, 6H), 1.05 (s, 9H).

Step 3: Preparation of 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionyl chloride (compound A4)

A solution of 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionic acid (compound A3, 163.0 g, 457.1 mmol) and DMF (166.8 mg, 2.3 mmol) in DCM (50 mL) was added with thionyl chloride (265.6 mL, 3657 mmol). The mixture was stirred at 50 °C for 12 hours. The mixture was concentrated under vacuum to give 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionyl chloride (compound A4, 171.4 g) as a yellow oil, which could be used in the next step without further purification.

Step 4: Preparation of 1-(5-bromo-1H-indol-3-yl)-3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-prop-1-one (compound A5)

At 0 °C, _SnCl₄ solution (140 mL, 140 mmol) was slowly added to a mixture of 3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethylpropionyl chloride (compound A4, 52.5 g, 140 mmol) in DCM (350 mL). The mixture was stirred at -10 °C for 0.5 h. Then, 5-bromoindole (27.4 g, 140 mmol) was added dropwise to DCM (150 mL). After addition, the mixture was stirred at -10 °C for 15 min. The mixture was diluted at 0 °C with saturated _NaHCO₃ aqueous solution (1000 mL) and then extracted twice with EtOAc (800 mL). The combined organic layers were washed with brine (600 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum. The residue was ground together with EtOAc (100 mL) to give 1-(5-bromo-1H-indol-3-yl)-3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-prop-1-one (compound A5, 40 g) as a yellow solid. ^¹H NMR (400 MHz, CDCl₃) δppm 8.68 (s, 1H), 8.60 (br. s, 1H), 7.67 (d, J = 2.8 Hz, 1H), 7.54–7.48 (m, 4H), 7.42–7.35 (m, 3H), 7.32–7.25 (m, 5H), 3.90 (s, 2H), 1.42 (s, 6H), 0.96 (s, 9H).

Step 5: Preparation of [3-(5-bromo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A6)

At 0 °C under _N2 protection, LiBH4 (28.1 mL, 112.2 mmol) was added to a solution of 1-(5-bromo-1H-indol-3-yl)-3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-prop- _1-one (compound A5, 20.0 g, 37.4 mmol) in THF (250 mL). The mixture was stirred at 60 °C for 12 hours. Once the reaction was complete, it was cooled to 25 °C and quenched with MeOH (20 mL). The mixture was then diluted with EtOAc (300 mL) and washed with brine (250 mL). The organic layer was dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated under vacuum. The residue was dissolved in DCM (250 mL) and cooled to 10 °C. Diethyl 1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylate (3.7 g, 14.9 mmol) and p-toluenesulfonic acid monohydrate (356.7 mg, 1.8 mmol) were added. After stirring at 10 °C for 2 hours, the mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-50%) to give [3-(5-bromo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A6, 17.2 g) as a colorless oil. ¹ HNMR (400MHz, CDCl3) δ8.00 (s, 1H), 7.75 (s, 1H), 7.74-7.65 (m, 4H), 7.47-7.36 (m, 6H), 7.2 6-7.19 (m, 2H), 6.89 (d, J=2.0Hz, 1H), 3.40 (s, 2H), 2.73 (s, 2H), 1.15 (s, 9H), 0.89 (s, 6H).

Step 6: Preparation of [3-(5-bromo-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A7)

Silver trifluoromethanesulfonate (24.4 g, 94.9 mmol) was added to a solution of [3-(5-bromo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A6, 41.2 g, 79.1 mmol) and iodine (20.1 g, 79.1 mmol) in THF (500 mL). After stirring at 25 °C for 1 hour, the mixture was quenched with a saturated aqueous solution _{of Na₂SO₃} (400 mL) and extracted with EtOAc (500 mL). The organic layer was washed with brine (400 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give a yellow solid [3-(5-bromo-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A7, 46 g). ^¹H NMR (400 MHz, CDCl₃) δppm 8.07 (s, 1H), 7.75–7.67 (m, 5H), 7.46–7.37 (m, 6H), 7.26–7.15 (m, 2H), 3.49 (s, 2H), 2.70 (s, 2H), 1.15 (s, 9H), 0.94 (s, 6H). MS: calculated 646 ( ^MH⁺ ); measured 645.9 ( ^MH⁺ ).

Step 7: Preparation of [3-[5-bromo-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A8)

Potassium carbonate (9.6 g, 69.6 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (1018.6 mg, 1.4 mmol) were added to a solution of [3-(5-bromo-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A7, 18 g, 27.8 mmol) in 1,4-dioxane (200 mL) and water (30 mL). The mixture was stirred at 80 °C for 12 hours under _N2 . The mixture was cooled to 20 °C, diluted with EtOAc (200 mL), and filtered. The filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give a brown oily substance, [3-[5-bromo-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A8, 10 g). ¹ H NMR (400MHz, CDCl3) δppm 9.31 (s, 1H), 8.32 (dd, J=4.4, 0.8Hz, 1H), 7.83-7.76 (m, 2H), 7.64-7.58 (m, 4H), 7.46-7.34 (m, 7H), 7.32-7.29 (m, 1H), 7.26-7.23 (m, 1H) , 4.52 (q, J=6.4Hz, 1H), 3.34 (s, 3H), 3.33-3.27 (m, 2H), 2.90-2.80 (m, 2H), 1.41 (d, J=6.4Hz, 3H), 1.07 (s, 9H), 0.64 (s, 3H), 0.59 (s, 3H). MS: Calculated value 657 (MH ⁺ ); Measured value 657.0 (MH ⁺ ).

Step 8: Preparation of [3-[5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A9)

At 0 °C, cesium carbonate (35.7 g, 109.8 mmol) and iodoethane (8.7 mL, 109.8 mmol) were added to a solution of [3-[5-bromo-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A8, 36.0 g, 54.9 mmol) in DMF (300 mL). After stirring at 25 °C for 12 hours, the mixture was diluted with water (1000 mL) and extracted with EtOAc (500 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a brown oily substance, [3-[5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound A9, 32 g). MS: calculated value 685 (MH ⁺ ); measured value 685.0 (MH ⁺ ).

Step 9: Preparation of 3-[5-bromo-1-ethyl-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-propane-1-ol (Intermediate A)

Tetrabutylammonium fluoride (280.7 mL, 280.7 mmol, 1 M, in THF) was added to a solution of [3-[5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]tert-butyl-diphenyl-silane (compound A9, 32.0 g, 46.8 mmol) in THF (200 mL). After stirring at 50 °C for 12 hours, the mixture was diluted with water (500 mL) and extracted with EtOAc (300 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give 3-[5-bromo-1-ethyl-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol (intermediate A, 7.4 g, fast elution) as a yellow gel and 3-[5-bromo-1-ethyl-(2P)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol (compound A10, 5.6 g, slow elution) as a yellow solid. Intermediate A: ^1H NMR (400 MHz, _CDCl₃ ) δppm 8.83 (dd, J=4.8, 2.0Hz, 1H), 7.90 (d, J=1.6Hz, 1H), 7.69 (dd, J=7.6, 1.6Hz, 1H), 7.39- 7.32(m, 2H), 7.26-7.23(m, 1H), 4.12-4.07(m, 1H), 4.03-3.95(m, 1H), 3.92-3.81(m, 1 3.27 (dd, J = 24.810.4 Hz, 2H), 3.07 (s, 3H), 2.74 (d, J = 14.0 Hz, 1H), 2.26 (d, J = 14.0 Hz, 1H), 1.60-1.52 (m, 1H), 1.48 (d, J = 6.4 Hz, 3H), 1.19 (t, J = 7.2 Hz, 3H), 0.77 (s, 6H). MS: Calculated value 445 (MH ⁺ ); Measured value 445.1 (MH ⁺ ).

X-ray crystallographic analysis of intermediate A

The absolute configuration of intermediate A was confirmed by X-ray crystallographic analysis of its single crystal. (Figure 1)

Intermediate B

(3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester

Intermediate B was prepared according to the following scheme:

Step 1: Preparation of methyl (2S)-2-(-3-bromophenyl)-3-(methoxycarbonylamino)propionate (compound B2)

Thionyl chloride (10 mL, 137.9 mmol) was added to a solution of LM-tyrosine (compound B1, 5.0 g, 27.6 mmol) in methanol (80 mL). The mixture was stirred at 60 °C for 12 h. The reaction mixture was cooled to 20 °C and concentrated under vacuum to give methyl (2S)-2-amino-3-(3-hydroxyphenyl)propionate (compound B2, 6.2 g) as a yellow solid. ^¹H NMR (400 MHz, CD₃OD) δppm 7.18 (t, J = 8.0 Hz, 1H), 6.78–6.66 (m, 3H), 4.29 (t, J = 6.4 Hz, 1H), 3.82 (s, 3H), 3.23–3.05 (m, 2H).

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

Sodium bicarbonate (40.6 g, 483.4 mmol) was added to a solution of methyl (2S)-2-amino-3-(3-hydroxyphenyl)propionate (compound B2, 32.0 g, 138.1 mmol) in THF (80 mL) and water (20 mL) at 20 °C, followed by di-tert-butyl dicarbonate (33.1 g, 151.9 mmol). The mixture was stirred at 20 °C for 12 hours. The mixture was diluted with water (100 mL) and acidified with 1 M HCl aqueous solution until pH = 5. The mixture was extracted three times with ethyl acetate (100 mL). The combined organic phases were washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give methyl (2S)-2-(tert-butoxycarbonylamino)-3-(3-hydroxyphenyl)propionate (compound B3, 40 g) as a colorless gel. MS: Calculated value 318 (MNa ⁺ ); Measured value 318.3 (MNa ⁺ ).

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

At 0 °C, triisopropylchlorosilane (39.1 g, 203.1 mmol) was added dropwise to a solution of (2S)-2-(tert-butoxycarbonylamino)-3-(3-hydroxyphenyl)propionate (compound B3, 40.0 g, 135.4 mmol) and 1H-imidazole (27.6 g, 406.3 mmol) in DMF (400 mL). After stirring at 25 °C for 12 hours, the mixture was diluted with water (250 mL) at 0 °C and extracted three times with ethyl acetate (200 mL). The combined organic phases were washed four times with brine (80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give methyl (2S)-2-(tert-butoxycarbonylamino)-3-(3-triisopropylsilyloxyphenyl)propionate (compound B4, 60 g) as a yellow oil. MS: calculated value 474 (MNa ⁺ ); measured value 474.2 (MNa ⁺ ).

Step 4: Preparation of methyl (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionate (compound B5)

[Ir(OMe)(COD)]₂ (2.2 g, 3.3 mmol) was added to a solution of (2S)-2-(tert-butoxycarbonylamino)-3-(3-triisopropylsilyloxyphenyl)propionate (compound B4, 15.0 g, 33.2 mmol), 4,4′-di-tert-butyl-2,2'-bipyridine (2.6 g, 9.9 mmol), and bis(pinacol)diboron (12.6 g, 49.8 mmol) in hexane (200 mL). The mixture was degassed and purged _three times with N₂. The resulting mixture was stirred at 70 °C for 12 hours. The reaction mixture was then cooled to 20 °C, diluted with petroleum ether (100 mL), and filtered. The filtrate was concentrated under vacuum to obtain a residue, which was purified by rapid chromatography (EA/PE: 0-20%) to give (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionate (compound B5, 21 g), a yellow oil. MS: calculated value 600 (MNa ⁺ ); measured value 600.3 (MNa ⁺ ).

Step 5: Preparation of (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionic acid (compound B6)

A solution of lithium hydroxide (3.2 mL, 346.2 mmol) in water (100 mL) was added to a solution of (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionate (compound B5, 40.0 g, 69.2 mmol) in methanol (300 mL). After stirring at 20 °C for 1 hour, the reaction mixture was diluted with water (200 mL) and the MeOH was removed under vacuum. The resulting mixture was acidified with 1 M HCl aqueous solution until pH 5. The resulting mixture was extracted three times with EtOAc (250 mL). The organic phase was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionic acid (compound B6, 33 g) as a white solid. MS: calculated value 586 (MNa ⁺ ); measured value 586.3 (MNa ⁺ ).

Step 6: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (intermediate B)

N,N-diisopropylethylamine (6.4 g, 49.6 mmol) was added to a solution of (2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionic acid (compound B6, 8.0 g, 14.1 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylurea hexafluorophosphate (5.6 g, 14.9 mmol) in DMF (100 mL). The mixture was stirred at 0 °C for 10 min. Then (3S)-hexahydropyridazine-3-carboxylate hydrochloride (compound B7, 2.6 g, 14.9 mmol) was added. The resulting mixture was stirred at 20 °C for 1.5 h. The mixture was diluted with water (200 mL) and extracted twice with EtOAc (100 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give methyl (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylate (intermediate B, 7.8 g) as a yellow oil. MS: calculated value 690 (MH ⁺ ); measured value 690.4 (MH ⁺ ).

Intermediate C

(8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione

Intermediate C was prepared according to the following scheme:

Step 1: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-5-yl]-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound C1)

Pd(dtbpf)Cl₂ (87.7 mg, 0.13 mmol) was added to a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylate (intermediate B, 1.1 g, 1.6 mmol) and 3-[5-bromo-1-ethyl-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol- _3- yl]-2,2-dimethyl-prop-1-ol (intermediate A, 356.9 mg, 3.3 mmol) in 1,4-dioxane (12 mL) and water (1.2 mL). The mixture was degassed and purged three times with _N₂ . The resulting mixture was stirred at 85°C for 12 hours. The reaction mixture was cooled to 20°C and filtered. The filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-50%) to give a yellow oil (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-5-yl]-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound C1, 750 mg). MS: calculated value 928 (MH ⁺ ); measured value 928.3 (MH ⁺ ).

Step 2: Preparation of (3S)-1-[(S)-2-(tert-butoxycarbonylamino)-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-5-yl]-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound C2)

Trimethyltin hydroxide (519.5 mg, 2.8 mmol) was added to a solution of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-5-yl]-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylate (compound C1, 750.0 mg, 0.7 mmol) in DCE (12 mL). The mixture was stirred at 60 °C for 12 h. LCMS showed that the starting material was consumed and the desired product was formed. The mixture was added to water (40 mL) and extracted three times with ethyl acetate (50 mL). The combined organic phases were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-5-yl]-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound C2, 650 mg), as a yellow solid. MS: calculated value 914 (MH ⁺ ); measured value 914.5 (MH ⁺ ).

Step 3: Preparation of N-[(8S,14S)-22-ethyl-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]tert-butyl carbamate (compound C3)

At 0 °C, DIEA (1.4 g, 11.3 mmol), EDCI (1.9 g, 10.6 mmol), and then HOBT (240.1 mg, 1.8 mmol) were added to a solution of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-5-yl]-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound C2, 650.0 mg, 0.7 mmol) in DCM (10 mL). The mixture was stirred at 20 °C for 12 hours. The mixture was poured into water (40 mL) and extracted three times with EtOAc (30 mL). The combined organic phases were washed with brine (40 mL), dried over sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-50%) to give N-[(8S,14S)-22-ethyl-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecano-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound C3, 530 mg). MS: Calculated value 896 (MH ⁺ ); Measured value 896.2 (MH ⁺ ).

Step 4: Preparation of N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]tert-butyl carbamate (compound C4)

At 0 °C, TBAF (0.6 mL, 0.6 mmol, 1.0 M, in THF) was added to a solution of N-[(8S, 14S)-22-ethyl-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecano-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound C3, 480.0 mg, 0.5 mmol) in THF (5 mL). After stirring at 0°C for 0.5 hours, the reaction mixture was diluted with water (30 mL) and extracted three times with EtOAc (40 mL). The combined organic phases were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated under vacuum to give N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound C4, 390 mg) as a colorless gel. MS: Calculated value 740 (MH ⁺ ); Measured value 740.2 (MH ⁺ ).

Step 5: Preparation of (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C)

TFA (0.8 mL) was added to a solution of N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound C4, 430.0 mg, 0.6 mmol) in DCM (4 mL). After stirring at 20 °C for 1 hour, the reaction mixture was diluted with saturated _NaHCO3 aqueous solution (40 mL) and extracted three times with EtOAc (60 mL). The combined organic phases were washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated under vacuum to give a yellow solid (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C, 330 mg). MS: calculated 640 (MH ⁺ ); measured 640.3 (MH ⁺ ). ^¹H NMR (400 MHz, _CD₃OD ) δppm 8.72-8.71 (m, 1H), 7.89 (d, J=7.6Hz, 1H), 7.81 (s, 1H), 7.55-7.44 (m, 3H), 7.11 (s, 1H), 6.92 (s, 1H), 6.60 (s, 1H), 4.7 5 (t, J=7.2Hz, 1H), 4.60 (br.s, 1H), 4.41 (d, J=12.8Hz, 1H), 4.35-4.20 (m, 2H), 4.14-4.05 (m, 1H), 3.62 (d, J=10.8Hz, 1 H), 3.49 (d, J=10.8Hz, 1H), 3.26 (s, 2H), 3.13-3.00 (m, 1H), 2.89-2.84 (m, 1H), 2.81-2.78 (m, 3H), 2.66-2.62 (m, 1H), 2.05-2.00 (m, 1H), 1.82-1.80 (m, 1H), 1.51-1.46 (m, 5H), 1.02 (t, J=6.8Hz, 3H), 0.86-0.82 (m, 3H), 0.59-0.56 (m, 3H).

Intermediate D4-[(5M)-5-[5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)indol-2-yl]-6-[(1S)-1-[methoxyethyl]-3-pyridyl]piperazine]-1-carboxylic acid benzyl ester

The title compound was prepared in a manner similar to that used in the preparation of intermediate A, by using 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D5) instead of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (compound A11).

Compound D5 was prepared according to the following scheme:

Step 1: Preparation of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)pyridine (compound D2)

Add 4,4′-di-tert-butyl-2,2′-bipyridine (372.7 mg, 1.39 mmol) and [Ir(OMe)(COD)] _₂ (306.3 mg, 0.460 mmol) to a solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (compound D1, 2.0 g, 9.26 mmol) and bis(pinacol)diboron (3.5 g, 13.9 mmol) in THF (30 mL). Stir the mixture at 75 °C for 16 hours under _N₂ . Filter the mixture and concentrate the filtrate under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)pyridine (compound D2, 2.4 g), which was a yellow oil. ^¹H NMR (400 MHz, CDCl₃) δppm 8.91 (d, J = 1.4 Hz, 1H), 8.21 (d, J = 1.4 Hz, 1H), 4.95 (q, J = 6.5 Hz, 1H), 3.30 (s, 3H), 1.49 (d, J = 6.5 Hz, 3H), 1.35 (s, 12H).

Step 2: Preparation of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (compound D3)

N-iodosuccinimide (4.1 g, 18.27 mmol) was added to a solution of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron-2-yl)pyridine (compound D2, 2.5 g, 7.3 mmol) in ACN (40 mL). The mixture was stirred at 90 °C for 40 h under _N2 . The mixture was quenched with saturated _Na2SO3 solution ( ₄₀ mL) and extracted twice with EtOAc (30 mL). 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 D3, 660 mg) as a yellow oil. MS calculated value 342 (MH ⁺ ); measured value 341.8 (MH ⁺ ).

Step 3: Preparation of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D5)

Cesium carbonate (1.6 g, 4.83 mmol), (R)-BINAP (60.1 mg, 0.1 mmol), and palladium(II) acetate (43.3 mg, 0.19 mmol) were added to a solution of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (compound D3, 660 mg, 1.9 mmol) and 1-Cbz-piperazine (compound D4, 425.1 mg, 1.9 mmol) in toluene (10 mL). The mixture was stirred at 100 °C for 12 hours under N2. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-50%) to give benzyl 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound D5, 740 mg) as a yellow solid. MS calculated value 434.1 (MH ⁺ ); measured value 434.1 (MH ⁺ ).

Intermediate E

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

Intermediate E was prepared according to the following scheme:

Step 1: Preparation of (4-bromothiazol-2-yl)methanol (compound E2)

Sodium borohydride (1.77 g, 46.87 mmol) was added to a solution of 4-bromothiazol-2-carboxaldehyde (6.0 g, 31.25 mmol) in methanol (70 mL) at 0 °C. The mixture was stirred at 25 °C for 1 hour. The reaction mixture was quenched with water (300 mL) at 0 °C and extracted three times with ethyl acetate (200 mL). 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 E2, 6 g) as a colorless oil.

Step 2: Preparation of 4-bromo-2-(bromomethyl)thiazole (compound E3)

At 0 °C, _CBr₄ (15.38 g, 46.38 mmol) and triphenylphosphine (12.16 g, 46.38 mmol) were added to a solution of (4-bromothiazol-2-yl)methanol (compound E2, 6.0 g, 30.92 mmol) in DCM (80 mL). After stirring at 25 °C for 1 hour, the mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in petroleum ether (0–10%) to give (4-bromothiazol-2-yl)methanol (compound E3, 6.0 g) as a yellow oil. MS calculated value 255.9 ( ^MH⁺ ); measured value 255.9 ( ^MH⁺ ).

Step 3: Preparation of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (compound E5)

At -78°C, n-butyllithium (10 mL, 25.22 mmol, 2.5 M) was slowly added to a mixture of (R)-2,5-dihydro-3,6-dimethoxy-2-isopropylpyrazine (compound E4, 4.32 g, 23.45 mmol) in THF (60 mL). After addition, the mixture was stirred at -78°C for 0.5 h. At -78°C, 4-bromo-2-(bromomethyl)thiazole (compound E3, 5.4 g, 21.02 mmol) was added to the above mixture, and the mixture was stirred for another 1 h. The mixture was quenched with saturated _NH4Cl aqueous solution (100 mL) and extracted twice with EtOAc (100 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by reversed-phase chromatography, eluting with _H₂O (0.01% FA) containing ACN at a concentration of 0–60% to give a yellow oily substance, 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (compound E5, 3.6 g). MS calculated value 360 (MH ⁺ ), measured value 359.9 (MH ⁺ ).

Step 4: Preparation of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound E6)

Hydrochloric acid (66.62 mL, 0.3 M) was added to a solution of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (compound E5, 3.6 g, 10 mmol) in ACN (20 mL). The mixture was stirred at 25 °C for 2 h. The mixture was alkalized with a saturated aqueous solution _{of NaHCO3} until pH 8. The mixture was extracted twice with EtOAc (80 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound E6, 3.1 g) as a yellow oil. MS calculated value 264.9 (MH ⁺ ), measured value 264.9 (MH ⁺ ).

Step 5: Preparation of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound E7)

Triethylamine (2.96 g, 29.23 mmol) and (Boc) _₂O (3.83 g, 17.54 mmol) were added to a solution of (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (E6, 3.1 g, 11.69 mmol) in DCM (40 mL). The mixture was stirred at 30 °C for 12 h. The mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate (0–30%) in petroleum ether to give methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound E7, 3.2 g) as a yellow oil. MS calculated value 387 ( ^MNa⁺ ); measured value 386.9 ( ^MNa⁺ ).

Step 6: Preparation of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionic acid (compound E8)

Lithium hydroxide (0.41 mL, 43.81 mmol) was added to a solution of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound E7, 3.2 g, 8.76 mmol) in THF (30 mL), methanol (2 mL), and water (10 mL). The mixture was stirred at 25 °C for 1 hour. The mixture was acidified with 1 M HCl aqueous solution until pH 5. The mixture was extracted twice with EtOAc (40 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound E8, 3.1 g) as a yellow oil. MS calculated value 373 (MNa ⁺ ); measured value 372.9 (MNa ⁺ ).

Step 7: Preparation of (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (E)

At 0 °C, (3S)-hexahydropyridazine-3-carboxylic acid methyl ester hydrochloride (compound E9, 2.39 g, 13.24 mmol), EDCI (3.38 g, 17.65 mmol), 1-hydroxybenzotriazole (238.53 mg, 1.77 mmol), and NMM (9.92 mL, 88.26 mmol) were added to a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionic acid (compound E8, 3.1 g, 8.83 mmol) in DCM (50 mL). The mixture was stirred at 25 °C for 1 hour. The mixture was diluted with water (60 mL) and extracted three times with EtOAc (60 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography and eluted with ethyl acetate in petroleum ether at 10–30% to give methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate E, 2.4 g). MS calculated value 477 (MH ⁺ ), measured value 476.9 (MH ⁺ ).

intermediate F

(7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione

Compound intermediate F was prepared according to the following scheme:

Step 1: 4-[(5M)-5-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound F3)

To a solution of 4-[(5M)-5-[5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (intermediate D, 110.0 mg, 0.17 mmol) and bis(pinacol)diboron (46.3 mg, 0.18 mmol) in toluene (4 mL), KOAc (40.67 mg, 0.4 mmol) and [1,1′-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (12.13 mg, 0.02 mmol) were added. After stirring at 90 °C for 12 hours under _N2 protection, the mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in petroleum ether at 30–60% to give a yellow oily compound, 4-[(5M)-5-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound F3, 100 mg). MS calculated value 711 (MH ⁺ ), measured value 711.1 (MH ⁺ ).

Step 2: Preparation of 4-[(5M)-5-[(7S,13S)-7-(tert-butoxycarbonylamino)-21-ethyl-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexen-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound F1)

By using 4-[(5M)-5-[1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid (compound F3) and (3S)-1-[(2S)-3-(4-bromothiazolyl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (intermediate E) instead of 3-[(5M) Compound F1 was prepared by means of intermediates A and B, namely (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (intermediate B).

Step 3: Preparation of N-[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexene-7-yl]tert-butyl carbamate (compound F2)

Add Pd(OH)₂/C (20 mg) to a solution of 4-[(5 M)-5-[(7 S, 13 S)-7-(tert-butoxycarbonylamino)-21-ethyl-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexen-20-yl]-6-[(1 S)-1-methoxyethyl]-3-pyridyl]piperazine- ₁ -carboxylic acid benzyl ester (compound F1, 35.0 mg, 0.04 mmol) in methanol (5 mL). Degas the mixture and purge it three times with _H₂ . After stirring at 25°C for 3 hours under an H2 balloon, the mixture was filtered, and the filtrate was concentrated under vacuum to obtain a white solid intermediate (20 mg). Acetic acid (5.04 mg, 0.08 mmol) was added to a solution of this intermediate (20.0 mg) in methanol (1 mL). After stirring at 25°C for 15 min, formaldehyde (5 mg, 0.06 mmol) and _NaBH3CN (2.11 mg, 0.03 mmol) were added to the mixture, followed by stirring for another 45 min. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by reversed-phase chromatography to give N-[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound F2, 17 mg). MS calculated value 829 (MH ⁺ ); measured value 829.1 (MH ⁺ ).

Step 4: Preparation of (7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate F)

Add TFA (1.0 mL) to a mixture of N-[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound F2, 17.0 mg, 0.02 mmol) in DCM (5 mL). After stirring at 30°C for 1 hour, the mixture was concentrated under vacuum and diluted with saturated _NaHCO3 solution (10 mL), and extracted with EtOAc. The combined organic layers were washed with brine (15 mL), filtered, and concentrated under vacuum to give a yellow oily substance (7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate F, 10 mg). MS calculated value 729 (MH ⁺ ), measured value 729.2 (MH ⁺ ).

intermediate G

(8S, 14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12, 6.110, 14.023, 27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione

Compounds were prepared according to the following scheme.

Step 1: Preparation of N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]tert-butyl carbamate (compound G2)

At 0 °C, TBAF (0.53 mL, 0.53 mmol, 1 M, in THF) was added to a solution of N-[(8S, 14S)-22-ethyl-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecano-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound G1, 500 mg) in THF (5 mL), and stirred for 0.5 h. After the reaction was complete, water (40 mL) was added, and the mixture was extracted three times with EtOAc (30 mL). The combined organic phases were washed four times with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound G2, 430 mg), which was a yellow gel. MS: Calculated value 838.5 (MH ⁺ ); Measured value 838.6 (MH ⁺ ).

Step 2: Preparation of (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate G)

TFA (0.8 mL) was added to a solution of N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound G2, 430 mg, 0.51 mmol) in DCM (4 mL). The mixture was stirred at 20 °C for 12 h. The mixture was added to a saturated aqueous solution of _NaHCO3 (30 mL) and extracted three times with EtOAc (20 mL). The combined organic phases were washed twice with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a yellow gelatinous substance (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate G, 360 mg). MS: Calculated value 738.4 (MH ⁺ ); Measured value 738.4 (MH ⁺ ).

N-[(8S,14S]methyl hexahydropyridazine-3-carboxylate (intermediate B) was prepared in a manner similar to that used for intermediate F, by replacing (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylate (intermediate E) with (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate E). 22-Ethyl-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]tert-butyl carbamate (compound G1).

intermediate H

(7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione

The title intermediate was prepared in a manner similar to that used in the preparation of intermediate F, by using 3-[5-bromo-1-ethyl-(2M)-2-[2-[(1S)-1-methoxyethyl]-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol (intermediate A) instead of 4-[5-[5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethyl-propyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester.

Intermediate I

(7S, 13S)-7-amino-(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[17.5.2.12, 5.19, 13.022, 26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione

The title intermediate was prepared in a manner similar to that used for _intermediate F, by using _CF3CH2OTf instead of iodoethane.

Example 1

(3S)-1-[(2S)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-NN-methyl-pyrrolidine-3-carboxamide

The title compound was prepared according to the following scheme:

Step 1: Preparation of N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate tert-butyl ester (compound 1b)

Add DIEA (90.9 mg, 0.7 mmol) to a solution of BOC-N-ME-VAL-OH (compound 1a, 65.1 mg, 0.3 mmol), (8S, 14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12, 6.110, 14.023, 27]nonadecane-1(26), 2,4,6(29), 20,23(27), 24-heptaene-9,15-dione (intermediate C, 150.0 mg, 0.23 mmol) in DMF (1.5 mL), and then HATU (55.1 mg, 0.23 mmol). After stirring at 0°C for 1 hour, the residue was diluted with water (40 mL) and extracted three times with EtOAc (60 mL). The combined organic phases were washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography and eluted with ethyl acetate in petroleum ether at 0–20% to give N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate tert-butyl ester (compound 1b, 180 mg), which was a yellow oil. MS: Calculated value 853 (MH ⁺ ); Measured value 853.2 (MH ⁺ ).

Step 2: Preparation of (2S)-N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-(methylamino)butyramide (compound 1c)

Add 0.4 mL of TFA to a solution of N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate tert-butyl ester (compound 1b, 180.0 mg, 0.19 mmol) in DCM (2 mL). After stirring at 20°C for 1 hour, the mixture was poured into a saturated _NaHCO3 aqueous solution (30 mL) and extracted three times with EtOAc (30 mL). The combined organic phases were washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated under vacuum to give (2S)-N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-(methylamino)butyramide (compound 1c, 150 mg), which was a yellow solid. MS: Calculated value 775 (MNa ⁺ ); Measured value 775.5 (MNa ⁺ ).

Step 3: Preparation of (3S)-3-[[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine-1-tert-butyl carboxylate (compound 1e)

Add O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylurea hexafluorophosphate (88.23 mg, 0.23 mmol) to a solution of (S)-1-BOC-pyrrolidine-3-carboxylic acid (compound 1d, 54.11 mg, 0.250 mmol) and N,N-diisopropylethylamine (0.12 mL, 0.68 mmol) in DMF (2 mL). The mixture was added to a solution of (2S)-N-[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-(methylamino)butyramide (compound 1c, 145.6 mg, 0.19 mmol) in DMF (3 mL). The resulting mixture was stirred at 20 °C for 0.5 h. The mixture was then quenched with water (40 mL) and extracted three times with ethyl acetate (40 mL). The combined organic phases were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography and eluted with ethyl acetate in petroleum ether at 30%–100% to give (3S)-3-[[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine-1-tert-butyl carboxylate (compound 1e, 140 mg). MS: Calculated value 950 (MH ⁺ ); Measured value 950.6 (MH ⁺ ).

Step 4: Preparation of (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (compound 1f)

Add 0.8 mL of TFA to a solution of (3S)-3-[[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine-1-carboxylate (compound 1e, 140.0 mg, 0.13 mmol) in DCM (4 mL). After stirring at 20°C for 0.5 h, the mixture was diluted with saturated _NaHCO3 aqueous solution (30 mL) and extracted three times with ethyl acetate (40 mL). The combined organic phases were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (compound 1f, 110 mg), which was a yellow solid. MS: Calculated value 850 (MH ⁺ ); Measured value 850.6 (MH ⁺ ).

Step 5: Preparation of (3S)-1-[(2S)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (Example 1)

At 0 °C, to (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6 (29), 20, 23(27), 24-[heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (compound 1f, 150.0 mg, 0.2 mmol) and (2S)-2-chloro-2-fluoro-acetic acid (compound 1g, 59.5 mg, 0.5 mmol) were added to a solution of DMF (5 mL) with N,N-diisopropylethylamine (137 mg, 1.1 mmol) and _T3P (168.4 mg, 0.26 mmol, 50% in EtOAc). After stirring at 20 °C for 12 hours, the mixture was diluted with water (20 mL) and extracted three times with ethyl acetate (20 mL). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by preparative HPLC to obtain (3S)-1-[(2S)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (Example 1, 51.1 g), which was a grayish-white solid. MS: Calculated value 944 (MH ⁺ ); Measured value 944.6 (MH ⁺ ). ^¹H NMR (400MHz, _CD₃OD ) δppm 8.74–8.73 (m, ¹H), 8.03 (s, ¹H), 7.91–7.88 (m, ¹H), 7.61 (d, J = 8.4 Hz, ¹H), 7.56–7.49 (m, 2H), 7.39 (s, 1H), 7.07–7.05 (m, 1H), 6.89–6.74 (m, 1H), 6.57–6.49 (m, 1H), 5.64–5.59 (m, 1H), 4.74 (d, J = 11.2 Hz, 1H), 4.47 (d, J = 11.2 Hz, 1H). 13.6Hz, 1H), 4.31-4.21(m, 2H), 4.05-3.96(m, 1H), 3.86-3.60(m, 8H), 3.23-3.18(m, 3H), 3.00-2.75(m, 8H), 2.39 -1.91 (m, 5H), 1.75-1.57 (m, 2H), 1.47 (d, J=6.4Hz, 3H), 1.10-1.03 (m, 3H), 0.95-0.79 (m, 9H), 0.71-0.64 (m, 3H).

Example 2

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

The title compound was prepared in a manner similar to that of Example 1 by using (2R)-2-chloro-2-fluoro-acetic acid instead of (2S)-2-chloro-2-fluoro-acetic acid (1 g of compound). Example 2 (21 mg) was obtained as a white solid. MS: calculated value 944 (MH ⁺ ); measured value 944.6 (MH ⁺ ). ¹ H NMR (400MHz, CD3OD) δ8.75-8.71 (m, 1H), 8.21-8.15 (m, 1H), 8.03 (d, J=5. 2Hz, 1H), 7.91-7.86(m, 1H), 7.63-7.59(m, 1H), 7.56-7.48(m, 2H), 7.38(s , 1H), 7.09-7.04 (m, 1H), 6.90-6.72 (m, 1H), 6.49 (d, J=9.2Hz, 1H), 5.65- 5.54 (m, 1H), 4.73 (s, 1H), 4.45 (d, J = 12.4Hz, 1H), 4.34-4.17 (m, 2H), 4.09 -3.99(m, 1H), 3.96-3.52(m, 8H), 3.26-3.15(m, 3H), 3.08-2.90(m, 4.5H) , 2.87-2.72(m, 3.5H), 2.40-2.24(m, 1H), 2.22-2.08(m, 2H), 2.02-1.90(m , 1H), 1.75-1.57 (m, 2H), 1.46 (d, J=6.0Hz, 3H), 1.32-1.24 (m, 0.5H), 1.12 -0.99(m, 3.5H), 0.98-0.89(m, 3H), 0.89-0.76(m, 6H), 0.73-0.58(m, 3H).

Example 3

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-formyl-N-methyl-cyclobutaneformamide

Compound 3e was prepared in a manner similar to that of Example 1 by using 3-(methoxymethylene)cyclobutanecarboxylic acid (compound 3d) instead of (2S)-2-chloro-2-fluoroacetic acid (compound 1 g). Example 3 (8 mg) was obtained as a yellow solid by conversion of compound 3e and preparative HPLC under acidic conditions. MS: calculated value 863 (MH ⁺ ); measured value 863.6 (MH ⁺ ). ^¹H NMR (400 MHz, _CD₃OD ) δ 8.78 (d, J = 4.8 Hz, 1H), 8.45 (d, J = 7.2 Hz, 1H), 8.04–7.92 (m). 1H), 7.65-7.63(m, 1H), 7.56-7.53(m, 1H), 7.39(s, 1H), 7.07(s, 1H), 6.51(s, 1H), 5.65-5.59(m, 1H), 4.74-4.65(m , 1H), 4.48-4.38(m, 3H), 4.35-4.24(m, 1H), 3.99-3.89(m, 1H), 3.83-3.75(m, 2H), 3.06-2.94(m, 3H), 2.90-2.83(m, 1H), 2.80-2.71(m, 6H), 2.45-2.36(m, 1H), 2.30-2.08(m, 7H), 1.94-1.87(m, 1H), 1.71-1.60(m, 2H), 1.47(d, J=6.0H z, 3H), 1.19 (s, 3H), 1.17-1.07 (m, 4H), 0.96-0.91 (m, 3H), 0.87-0.83 (m, 4H), 0.82-0.78 (m, 3H), 0.75-0.71 (m, 3H).

Compound 3d was prepared according to the following scheme:

Step 1: Preparation of methyl 3-(methoxymethylene)cyclobutanecarboxylate (compound 3c)

At 0 °C, t-BuOK (17.5 g, 156.09 mmol) was added to a solution of (methoxymethyl)triphenylphosphonium chloride (compound 3b, 53.5 g, 156.09 mmol) in THF (500 mL). After 0.5 hours, methyl 3-oxocyclobutane carboxylate (compound 3a, 10.0 g, 78.05 mmol) was added. After stirring at 70 °C for 2.5 hours, the mixture was acidified with 1 M HCl aqueous solution until pH 5-6, and extracted twice with EtOAc (300 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in petroleum ether (0-10%) to give methyl 3-(methoxymethylene)cyclobutane carboxylate (compound 3c, 2.5 g) as a pale yellow oil. ¹ H NMR (400MHz, DMSO-d ₆ ) δppm 5.83-5.81 (m, 1H), 3.70 (s, 3H), 3.56 (s, 3H), 3.17-3.14 (m, 1H), 2.99-2.96 (m, 2H), 2.92-2.85 (m, 2H).

Step 2: Preparation of 3-(methoxymethylene)cyclobutanecarboxylic acid (compound 3d)

Lithium hydroxide (3.1 mg, 128.1 mmol) was added to a solution of methyl 3-(methoxymethylene)cyclobutanecarboxylate (compound 3c, 2.0 g, 12.8 mmol) in THF (50 mL), methanol (10 mL), and water (10 mL). After stirring at 25 °C for 2 hours, the reaction mixture was diluted with water (30 mL) and extracted twice with DCM (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 3-(methoxymethylene)cyclobutanecarboxylic acid (compound 3d, 1.0 g) as a pale yellow oil. ¹ H NMR (400MHz, DMSO-d ₆ ) δppm 5.89-5.87 (m, 1H), 3.47 (s, 3H), 3.10-3.01 (m, 1H), 2.88-2.79 (m, 1H), 2.78-2.70 (m, 3H).

Example 4

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-ethynyl-N-methyl-cyclobutaneformamide

The title compound was prepared in a manner similar to that of Example 1 by using 3-ethynylcyclobutanecarboxylic acid instead of (S)-1-BOC-pyrrolidine-3-carboxylic acid (compound 1d). Example 4 (20 mg) was obtained as a brown solid. MS: calculated value 859 (MH ⁺ ); measured value 859.5 (MH ⁺ ). ^¹H NMR (400 MHz, _CD₃OD ) δppm 8.73 (dd, J = 1.6, 4.8 Hz, 1H), 8.15–8.10 (m, 1H), 8.01–7.99 (m). 1H), 7.90-7.87(m, 1H), 7.61-7.48(m, 3H), 7.37-7.35(m, 1H), 7.06-7.04(m, 1H), 6.51-6.48(m, 1H), 5.65-5.57(m, 1H), 4.71- 4.66 (m, 1H), 4.46-4.43 (m, 1H), 4.33-4.21 (m, 2H), 4.07-4.01 (m, 1H), 3.81-3.62 (m, 3H), 3.49-3.36 (m, 1H), 3.26-3.21 (m, 3H) , 3.07-2.87(m, 3H), 2.84-2.66(m, 6H), 2.65-2.38(m, 4H), 2.37-2.25(m, 1H), 2.23-2.03(m, 3H), 1.94-1.88(m, 1H), 1.71-1.56 (m, 2H), 1.47 (d, J=6.0Hz, 3H), 1.36-1.19 (m, 1H), 1.06-1.01 (m, 3H), 0.98-0.86 (m, 3H), 0.85-0.76 (m, 6H), 0.65-0.58 (m, 3H).

Example 5

(3S)-N－[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-(2-oxopropionyl)pyrrolidine-3-carboxamide

The title compound was prepared in a manner similar to that of Example 1 by using pyruvate instead of (2S)-2-chloro-2-fluoro-acetic acid (1 g of compound). Example 5 (8 mg) was obtained as a white solid. MS: Calculated value 920 (MH ⁺ ); Measured value 920.5 (MH ⁺ ). ^1H NMR (400 MHz, _CD3 OD) δppm 8.73 (dd, J = 1.2, 4.8 Hz, 1H), 8.03 (s, 1H), 7.90–7.88 (m, 1H), 7.62–7.60 (m, 1H), 7.56–7.49 (m, 2H), 7.39–7.37 (m, 1H), 7.07–7.05 (m, 1H), 6.65–6.45 (m, 1H) ,5.69-5.55(m,1H),4.75-4.70(m,1H),4.53-4.44(m,1H),4.35-4.15(m,2H) , 4.13-3.98(m, 1H), 3.93-3.82(m, 1H), 3.81-3.72(m, 3H), 3.68-3.55(m, 3H), 3.26-3.21(m, 3H), 3.05-2.95(m, 1H), 2.94-2.88(m, 3H), 2.85-2.72(m, 3H), 2.42-2.40(m, 2H), 2.36-2.21(m, 1H), 2.18-1.90(m, 4H), 1.76-1.55(m, 2H), 1 .47(d, J=6.0Hz, 3H), 1.36-1.19(m, 2H), 1.10-0.96(m, 4H), 0.95-0.93(m, 2H) , 0.91-0.86 (m, 1H), 0.84-0.79 (m, 5H), 0.76-0.71 (m, 1H), 0.65-0.58 (m, 3H).

Example 6

(3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-prop-2-ynylpyrrolidine-3-carboxamide

The title compound was prepared in a manner similar to that of Example 1 using (2S)-3-methyl-2-[methyl-[(3S)-1-prop-2-ynylpyrrolidine-3-carbonyl]amino]butyric acid (compound 6h) instead of BOC-N-ME-VAL-OH (compound 1a). Example 6 (27 mg) was obtained as a white solid. MS: calculated value 888 (MH ⁺ ); measured value 888.7 (MH ⁺ ). ^1H NMR (400 MHz, _CD3 OD) δppm 8.80-8.78(m, 1H), 8.09-7.92(m, 2H), 7.69-7.33(m, 4H), 7.09-7.05(m, 1H), 6.60-6.48(m, 1H), 5.6 3-5.36(m, 1H), 4.73-4.65(m, 1H), 4.44-4.21(m, 4H), 4.10-3.99(m, 2H), 3.84-3.38(m, 8H), 3.27-2 .97(m, 8H), 2.85-2.73(m, 6H), 2.20-2.11(m, 2H), 1.97-1.90(m, 1H), 1.70-1.58(m, 2H), 1.48(dd, J =2.0, 6.0Hz, 3H), 1.36-1.29 (m, 1H), 1.19 (t, J = 7.2Hz, 1H), 1.12-0.83 (m, 10H), 0.70-0.63 (m, 3H).

Compound 6h was prepared according to the following scheme:

Step 1: Preparation of (2S)-3-methyl-2-(methylamino)butyrate tert-butyl ester (compound 6b)

Pd/C (1.0 g, 10% purity) was added to a solution of (2S)-2-[benzyloxycarbonyl(methyl)amino]-3-methyl-butyrate tert-butyl ester (compound 6a, 10.0 g, 31.11 mmol) in ethyl acetate (150 mL). The mixture was degassed under vacuum and purged three times with _H₂ . The resulting mixture was stirred at 20 °C for 12 h under an _H₂ balloon. TLC (PE:EtOAc = 3:1) showed complete depletion of the starting material. The mixture was filtered, and the filtrate was concentrated under vacuum to give (2S)-3-methyl-2-(methylamino)butyrate tert-butyl ester (compound 6b, 5.2 g) as a colorless oil, which could be used in the next step without purification.

Step 2: Preparation of (3S)-3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine-1-carboxylic acid benzyl ester (compound 6d)

At 0 °C, DIEA (10.4 mL, 60.1 mmol) and T3P (19.1 g, 30.1 mmol, 50% in EtOAc) were added to a solution of (2S)-3-methyl-2-(methylamino)butyrate (compound 6b, 4.1 g, 22.07 mmol) and (3S)-1-benzyloxycarbonylpyrrolidine- ₃ -carboxylic acid (compound 6c, 5.0 g, 20 mmol) in ethyl acetate (50 mL). After stirring at 0 °C for 1 hour, the reaction mixture was poured into water (300 mL) and extracted twice with EtOAc (200 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with EtOAc = 30–50% in petroleum ether to give (3S)-3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine-1-carboxylic acid benzyl ester (compound 6d, 7g) as a colorless oil. MS: calculated value 419 (MH ⁺ ); measured value 419.2 (MH ⁺ ).

Step 3: Preparation of tert-butyl (2S)-3-methyl-2-[methyl-[(3S)-pyrrolidine-3-carbonyl]amino]butyrate (compound 6e)

Pd/C (0.4 g, 10% purity) was added to a solution of (3S)-3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine-1-carboxylic acid benzyl ester (compound 6d, 4.0 g, 9.56 mmol) in ethyl acetate (50 mL). The mixture was degassed under vacuum and purged three times with _H₂ . After stirring at 20 °C for 12 hours under an _H₂ balloon, the reaction mixture was filtered, and the filtrate was concentrated under vacuum to give (2S)-3-methyl-2-[methyl-[(3S)-pyrrolidine-3-carbonyl]amino]tert-butyl butyrate (compound 6e, 2.3 g) as a colorless oil. MS: calculated value 285 (MH ⁺ ); measured value 285.1 (MH ⁺ ).

Step 4: Preparation of (2S)-3-methyl-2-[methyl-(1-prop-2-ynylpyrrolidine-3-carbonyl)amino]tert-butyl butyrate (compound 6g)

To a solution of (2S)-3-methyl-2-[methyl(pyrrolidine-3-carbonyl)amino]butyrate tert-butyl ester (compound 6e, 100.0 mg, 0.350 mmol) and potassium carbonate (48.6 mg, 0.35 mmol) in MeCN (2.5 mL), propargyl bromide (compound 6e, 52.2 mg, 0.35 mmol) was added. After stirring at 25 °C for 1 hour, the reaction mixture was diluted with water (50 mL) and extracted twice with ethyl acetate (50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the residue. The residue was purified by reversed-phase rapid column chromatography, eluting with 15–35% ACN (0.1% TFA) in water to give tert-butyl (2S)-3-methyl-2-[methyl-(1-prop-2-ynylpyrrolidine-3-carbonyl)amino]butyrate (6 g, 70 mg of compound). MS: calculated value 323 (MH ⁺ ); measured value 322.9 (MH ⁺ ).

Step 5: Preparation of (2S)-3-methyl-2-[methyl-[(3S)-1-prop-2-ynylpyrrolidine-3-carbonyl]amino]butyric acid (compound 6h)

Tert-butyl (2S)-3-methyl-2-[methyl-[(3S)-1-prop-2-ynylpyrrolidine-3-carbonyl]amino]butyrate (compound 6 g, 70 mg, 0.22 mmol) in TFA (0.5 mL) was stirred at 25 °C for 1.5 h. The mixture was concentrated under vacuum to give (2S)-3-methyl-2-[methyl-[(3S)-1-prop-2-ynylpyrrolidine-3-carbonyl]amino]butyric acid (compound 6 h, 75 mg) as a pale yellow oil. MS: calculated value 267 (MH ⁺ ); measured value 267.1 (MH ⁺ ).

Example 7

(3S)-1-(cyanomethyl)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

The title compound was prepared in a manner similar to that of Example 6 by using 2-bromoacetonitrile instead of 3-bromoprop-1-yne (compound 6f). Example 7 (42 mg) was obtained as a white solid. MS: calculated value 889 (MH ⁺ ); measured value 889.6 (MH ⁺ ). ^1H NMR (400 MHz, _CD3 OD) δppm 8.85-8.82(m, 1H), 8.21-8.17(m, 1H), 8.07-8.03(m, 1H), 7.79-7.75(m, 1H), 7.66-7.62(m, 1H), 7.56-7.54(m, 1H), 7.10-7.06(m, 1 H), 7.44-7.41(m, 1H), 6.67-6.64(0.6H), 6.50(s, 0.4H), 5.65-5.59(m, 1H), 4.73-4.67(m, 1H), 4.50-4.25(m, 6H), 4.04-3.97(m, 1 H), 3.84-3.66 (m, 5H), 3.50-3.40 (m, 2H), 3.14-2.73 (m, 11H), 2.60-2.48 (m, 1H), 2.25-2.09 (m, 3H), 1.98-1.90 (m, 1H), 1.78-1.59 (m, 2H), 1.48 (d, J=6.4Hz, 3H), 1.33-1.29 (m, 1H), 1.13-1.06 (m, 3H), 0.95-0.93 (m, 1H), 0.92-0.85 (m, 7H), 0.71 (d, J=6.8Hz, 3H).

Example 8

3-[2-[dimethyl(oxo) ^-λ6 -thionyl]acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-cyclobutaneformamide

The title compound was prepared in a manner similar to that of Example 1 using (2S)-2-[[3-[2-[dimethyl(oxo)-λ6-thionyl]acetyl]cyclobutanecarbonyl]-methyl-amino]-3-methyl-butyric acid (compound 8f) instead of BOC-N-ME-VAL-OH (compound 1a). Example 8 (8 mg) was obtained as a yellow solid. MS: calculated 953 (MH ⁺ ); measured 953.6 (MH ⁺ ). ^¹H NMR (400 MHz, _CD₃OD ) δppm 8.77 (d, J=4.8Hz, 1H), 8.03-7.97 (m, 2H), 7.64-7.58 (m, 2H), 7.53 (d, J=10.4Hz, 1H), 7.40-7.37 (m, 1H), 7.06-7.03 (m, 1H), 6.64-6.50 (m, 1H), 5.67-5.59(m, 1H), 4.73-4.75(m, 1H), 4.48-4.41(m, 1H), 4.34-4.22(m, 2H), 4.10-4.00(m, 1H), 3.94-3.87(m, 4H), 3.79-3.75(m, 1H) , 3.65-3.45(m, 5H), 3.25-3.21(m, 3H), 3.08-2.70(m, 9H), 2.65-2.51(m, 3H), 2.20-2.05(m, 2H), 1.95-1.88(m, 1H), 1.71-1.57(m, 2H), 1. 48 (d, J=6.0Hz, 3H), 1.34-1.27 (m, 1H), 1.19 (t, J=7.2Hz, 1H), 1.10-1.04 (m, 3H), 0.95-0.94 (m, 1H), 0.88-0.82 (m, 8H), 0.72-0.64 (m, 3H).

Compound 8f was prepared according to the following scheme:

Step 1: Preparation of methyl 3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]cyclobutanecarboxylate (compound 8c)

At 0 °C, DIEA (2.7 mL, 16.02 mmol) and T3P (5.1 g, 8.01 mmol, 50% in EtOAc) were added to a solution of (2S)-3-methyl-2-(methylamino)butyrate tert-butyl ester (compound 8b, ₁ g, 5.3 mmol) in ethyl acetate (15 mL). After stirring at 0 °C for 1 hour, the mixture was poured into water (50 mL) and extracted twice with EtOAc (50 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography and eluted with petroleum ether: EtOAc = 15-35% to give methyl 3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]cyclobutanecarboxylate (compound 8c, 740 mg) as a colorless oil. ¹ H NMR (400MHz, CD ₃ OD): δppm 4.57(d, J=10.4Hz, 0.6Hz), 3.65(s, 3H), 3.52-3.43(m, 1H), 2.96-3.05(m, 1H), 2.83-2.72(m, 3H) , 2.44-2.33 (m, 4H), 2.19-2.10 (m, 1H), 1.42 (s, 9H), 0.97 (d, J=6.4Hz, 3H), 0.80 (d, J=6.4Hz, 3H).

Step 2: Preparation of 3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]cyclobutanecarboxylic acid (compound 8d)

At 0 °C, NaOH (171 mg, 4.2 mmol) was added to a solution of methyl 3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]cyclobutanecarboxylic acid (compound 8c, 700 mg, 2.14 mmol) in THF (6 mL) and water (1.5 mL). After stirring at 20 °C for 2 hours, the mixture was poured into a 1 M HCl aqueous solution (50 mL) and extracted twice with EtOAc (50 mL). The combined organic phases were washed with brine (50 mL) and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give 3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-methyl-carbamoyl]cyclobutanecarboxylic acid (compound 8d, 670 mg) as a yellow oil. MS: Calculated value 314 (MH ⁺ ); Measured value 314.1 (MH ⁺ ).

Step 3: Preparation of (2S)-2-[[3-[2-[dimethyl(oxo)-λ6-thionyl]acetyl]cyclobutanecarbonyl]-methyl-amino]-3-methyl-butyrate tert-butyl ester (compound 8e)

t-BuOK (214.4 mg, 1.9 mmol) was added to a solution of trimethyl sulfoxide (421.3 mg, 1.9 mmol) in THF (3 mL) to form mixture A. Mixture A was stirred at 65 °C for 2 hours under _N2 protection and then cooled to 0 °C. In another flask, HATU (266.7 mg, 0.7 mmol) and Et3N (96.6 mg, 0.96 mmol) were added to 3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]methyl-carbamoyl]cyclobutanecarboxylic acid (compound 8d, 200 mg, 0.64 mmol) in THF ( ₃ mL) to form mixture B. Mixture B was stirred at 25 °C for 2 hours. Mixture B was then added to mixture A at 0 °C. The resulting reaction mixture was stirred at 5 °C for another 4 hours and then concentrated under vacuum. The residue was purified by preparative HPLC to give (2S)-2-[[3-[2-[dimethyl(oxo) ^-λ6 -thionyl]acetyl]cyclobutanecarbonyl]-methyl-amino]-3-methyl-butyrate tert-butyl ester (compound 8e, 60 mg) as a colorless oil. MS: calculated value 388 (MH ⁺ ); measured value 388.0 (MH ⁺ ).

Step 4: Preparation of (2S)-2-[[3-[2-[dimethyl(oxo) ^-λ6 -thionyl]acetyl]cyclobutanecarbonyl]-methyl-amino]-3-methyl-butyric acid (compound 8f)

TFA (1 mL) was added to a solution of (2S)-2-[[3-[2-[dimethyl(oxo) ^-λ6 -thionyl]acetyl]cyclobutanecarbonyl]-methyl-amino]-3-methyl-butyrate tert-butyl ester (compound 8e, 20 mg, 0.05 mmol) in DCM (3 mL). The mixture was stirred at 20 °C for 2 hours. The mixture was concentrated under vacuum to give (2S)-2-[[3-[2-[dimethyl(oxo) ^-λ6 -thionyl]acetyl]cyclobutanecarbonyl]-methyl-amino]-3-methyl-butyric acid (compound 8f, 17 mg) as a yellow oil. MS observations: Calculated value 331.8 (MH ⁺ ); Measured value 331.9 (MH ⁺ ).

Example 9

(3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-[(E)-3-(1,2,4-triazol-1-yl)propyl-2-enoyl]pyrrolidine-3-carboxamide

Compounds were prepared according to the following scheme.

Step 1: Preparation of (2S)-2-[[(3S)-1-[(E)-3-iodoprop-2-enoyl]pyrrolidine-3-carbonyl]-methyl-amino]-3-methyl-butyrate tert-butyl ester (compound 9b)

To a solution of (E)-3-iodoprop-2-enoic acid (compound 9e, 250.0 mg, 1.26 mmol) in DMF (4 mL), tert-butyl (2S)-3-methyl-2-[methyl-[(3S)-pyrrolidine-3-carbonyl]amino]butyrate (compound 9a, 359 mg, 1.26 mmol), DIEA (488.74 mg, 3.79 mmol), and _T3P (1044 mg, 1.64 mmol) were added. After stirring at 25 °C for 3 hours, the mixture was diluted with EtOAc (50 mL) and poured into water (30 mL). After separation, the aqueous solution was extracted three times with EA (30 mL). The combined organic phases were _dried over _Na2SO4 . After filtration and concentration, the residue was purified by silica gel column chromatography, eluting with PE:EA = 1:10 to give (2S)-2-[[(3S)-1-[(E)-3-iodoprop-2-enoyl]pyrrolidine-3-carbonyl]-methyl-amino]-3-methyl-butyrate tert-butyl ester (compound 9b, 350 mg) as a colorless oil. MS calculated value 465.3 (MH ⁺ ); measured value 465.2 (MH ⁺ ).

Step 2: Preparation of tert-butyl (2S)-3-methyl-2-[methyl-[(3S)-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carbonyl]amino]butyrate (compound 9c)

DABCO (72 mg, 0.65 mmol) was added to a solution of (2S)-2-[[(3S)-1-[(E)-3-iodoprop-2-enoyl]pyrrolidine-3-carbonyl]-methyl-amino]-3-methyl-butyrate tert-butyl ester (compound 9b, 150 mg, 0.32 mmol) in DMF (3 mL). After stirring at 25 °C for 10 min, 1,2,4-triazole (33 mg, 0.48 mmol) was added to the mixture, and the resulting mixture was stirred at 25 °C for 16 min. The mixture was diluted with EtOAc (50 mL) and poured into water (30 mL). The aqueous phase was extracted three times with EtOAc (30 mL). The combined organic phases were washed three times with EtOAc (30 mL), _dried over _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (MeOH = 0–5% in DCM) to give tert-butyl (2S)-3-methyl-2-[methyl-[(3S)-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carbonyl]amino]butyrate (compound 9c, 40 mg), a yellow oil. MS calculated value 406.2 (MH ⁺ ); measured value 406.1 (MH ⁺ ).

Step 3: Preparation of (2S)-3-methyl-2-[methyl-[(3S)-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carbonyl]amino]butyric acid (compound 9d)

TFA (1.0 mL) was added to a solution of (2S)-3-methyl-2-[methyl-[(3S)-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carbonyl]amino]butyrate (compound 9c, 30 mg, 0.07 mmol) in DCM (4 mL). After stirring at 20 °C for 1 hour, the mixture was concentrated under vacuum to give (2S)-3-methyl-2-[methyl-[(3S)-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carbonyl]amino]butyrate (compound 9d, 34 mg) as a yellow oil. MS calculated value 350.2 (MH ⁺ ); measured value 350.0 (MH ⁺ ).

Step 4: (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carboxamide (Example 9)

Add DIEA (16 mg, 0.13 mmol) and HATU (15 mg, 0.04 mmol) to a solution of (2S)-3-methyl-2-[methyl-[(3S)-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carbonyl]amino]butyric acid (compound 9d, 10 mg, 0.03 mmol) in DMF (2 mL). After stirring for 10 minutes, add (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.1]] to the mixture. [10, 14.023, 27] 26, 2, 4, 6 (29), 20, 23 (27), 24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carboxamide (20 mg, 0.03 mmol), and stirred for another 2 hours. After the reaction was complete, the mixture was purified by preparative HPLC to obtain (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapenta] as a white solid. Cyclo[18.5.2.12, 6.110, 14.023, 27]nonadecane-1(26), 2,4,6(29), 20,23(27), 24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carboxamide (Example 9, 8 mg). MS calculated value 971.5 (MH ⁺ ); measured value 971.7 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4 ) δppm 9.02-9.09(m, 1H), 8.86-8.90(m, 1H), 8.33-8.43(m, 1H), 8.08-8.12(m, 1H), 7.85-7.94(m, 1H), 7.62-7.69(m, 1H), 7.54-7.58(m, 1H), 7.39-7.44(m, 1H), 7.26-7.32(m, 1H), 7.03-7.07(m, 1H), 6.6-6.65(m, 1H), 6.07-6.15(m, 1H), 5 .59-5.65(m, 1H), 4.29-4.48(m, 1H), 3.94-4.02(m, 1H), 3.56-3.85(m, 8H), 2.90-3.06(m, 6H), 2.69-2.85(m, 4H), 2 .05-2.32(m, 4H), 1.88-1.96(m, 1H), 1.57-1.71(m, 2H), 1.43-1.52(m, 4H), 1.04-1.16(m, 4H), 0.73-0.98(m, 15H).

Example 10

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

By using (2R)-2-chloro-2-fluoro-acetic acid and (7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate F) instead of (2S)- The title compound was prepared by means of 2-chloro-2-fluoroacetic acid (compound 1 g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C) in a manner similar to that of Example 1. Example 10 (89 mg) was obtained as a yellow solid. MS calculated value 1033.5 (MH ⁺ ); measured value 1033.7 (MH ⁺ ). ^1H NMR (400 MHz, MeOD) δppm 8.61-8.54(m, 1H), 8.49(d, J=2.4Hz, 1H), 7.76-7.70(m, 2H), 7.59-7.49(m , 2H), 6.93-6.74(m, 1H), 5.85-5.67(m, 1H), 4.84-4.76(m, 1H), 4.47-4.31 (m, 3H), 4.27-4.19 (m, 1H), 4.18-4.04 (m, 2H), 3.98-3.86 (m, 1H), 3.85-3. 65(m,7H),3.64-3.45(m,4H),3.42-3.35(m,5H),3.28-3.23(m,1H),3.18- 3.11 (m, 1H), 3.10 (d, J = 4.0Hz, 3H), 2.99 (s, 3H), 2.80 (dt, J = 12.8, 2.8Hz, 1H), 2.69-2.56(m, 1H), 2.39-2.22(m, 3H), 2.20-2.02(m, 1H), 2.01-1.90( m, 1H), 1.88-1.72 (m, 1H), 1.69-1.55 (m, 1H), 1.45 (d, J=6.0Hz, 3H), 1.11- 1.01 (m, 1H), 1.01-0.93 (m, 9H), 0.87 (d, J=6.4Hz, 3H), 0.56-0.46 (m, 3H).

Example 11

(3S)-1-(bicyclo[1.1.0]butane-1-carbonyl)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

The compound was prepared according to the following scheme:

At 0 °C, sodium bicyclo[1.1.0]butane-1-carbonyloxy (compound 11a, 35 mg, 0.29 mmol), (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[ [18.5.2.12, 6.110, 14.023, 27] 1 (26), 2, 4, 6 (29), 20, 23 (27), 24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (compound 1f, 50 mg, 0.06 mmol) was added to a solution of DIEA (38 mg, 0.29 mmol) and _T3P (28 mg, 0.09 mmol) in DMF (1.5 mL). The mixture was stirred at 20 °C for 12 hours. After the reaction was complete, water (40 mL) was added and the mixture was extracted three times with EtOAc (30 mL). The combined organic phases were washed four times with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by preparative HPLC to obtain (3S)-1-(bicyclo[1.1.0]butane-1-carbonyl)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]] as a white solid. [-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (Example 11, 4 mg). MS calculated value 930.5 (MH ⁺ ); measured value 930.5 (MH ⁺ ). ¹ H NMR (400MHz, methanol-d4) δppm8.75-8.72 (m, 1H), 8.32 (s, 1H), 8.03 (s, 1H), 7.93-7.86 (m, 1H), 7.62-7.58 (m, 1H), 7.56-7.53 (m, 1H), 7.52 -7.49 (m, 1H), 7.41-7.33 (m, 1H), 7.10-7.04 (m, 1H), 6.49 (s, 1H), 5.66-5.55 (m, 1H), 5.45-5.22 (m, 1H), 4.78-4.72 (m, 2H), 4.46 (d, J =13.2Hz, 1H), 4.34-4.18(m, 2H), 3.84-3.68(m, 4H), 3.25-3.16(m, 3H), 3.00-2.90(m, 4H), 2.86-2.89(m, 4H), 2.33-2.23(m, 3H), 2.2 0-2.05 (m, 4H), 1.95-1.89 (m, 1H), 1.72-1.58 (m, 8H), 1.46 (d, J=6.8Hz, 3H), 1.40-1.34 (m, 7H), 0.84-0.82 (m, 3H), 0.68-0.60 (m, 3H).

Example 12

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

By using (2R)-2-chloro-2-fluoro-acetic acid and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate G) instead of (2S). The title compound was prepared in a manner similar to that of Example 1 using 2-chloro-2-fluoro-acetic acid (compound 1 g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C). Example 12 (17 mg) was obtained as a white solid. MS calculated value 1042.5 (MH ⁺ ); measured value 1042.7 (MH ⁺ ). ^¹H NMR (400MHz, methanol-d⁴) δppm: 8.42 (d, J = 3.2Hz, 1H), 8.04–8.00 (m, 1H), 7.62–7.58 (m, 1H), 7.49 (dd, J = 2.8, 8.8Hz, 1H), 7.41–7.34 (m, 2H), 7.09–7.04 (m, 1H), 6.90–6.73 (m, 1H), 6.57–6.47 (m, 1H), 5.64-5.55 (m, 1H), 4.72 (d, J=10.4Hz, 1H), 4.59 (s, 2H), 4.46 (d, J=13. 2Hz, 1H), 4.26-4.17(m, 2H), 4.12-4.07(m, 1H), 3.85-3.76(m, 3H), 3.73-3.64(m , 3H), 3.60-3.54(m, 1H), 3.39(t, J=4.4Hz, 4H), 3.22-3.14(m, 3H), 2.97-2.91( m, 4H), 2.84-2.79 (m, 2H), 2.78-2.73 (m, 5H), 2.44 (s, 3H), 2.40-2.27 (m, 1H), 2 .24-2.07(m, 3H), 2.02-1.91(m, 2H), 1.74-1.60(m, 2H), 1.43(d, J=6.4Hz, 3H), 1.11-1.05 (m, 3H), 0.94 (t, J=6.4Hz, 3H), 0.86-0.80 (m, 5H), 0.73-0.64 (m, 3H).

Example 13

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxamide

The compound was prepared according to the following scheme:

At 0 °C, (2S)-3-methyl-2-[methyl-[3-[2-(2-pyridyl)ethynyl]cyclobutanecarbonyl]amino]butyric acid (compound 13G, 20 mg, 0.06 mmol) was added to a solution of (8S, 14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C, 40 mg, 0.06 mmol) in DMF ( ₃ mL). P (60 mg, 0.1 mmol) was added, and then DIEA (41 mg, 0.32 mmol) was added slowly. After stirring at 20 °C for another 2 hours, the mixture was diluted with water (15 mL) and then extracted twice with EtOAc (15 mL). The combined organic phases were washed with brine (15 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain the residue. The residue was purified by preparative HPLC to give N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxamide (Example 13, 10.9 mg) as a white solid. MS: Calculated value 936.5 (MH+); Measured value 936.6 (MH+). ^¹H NMR (400MHz, methanol- _d⁴ ) δppm 8.73 (dd, J = 4.8, 1.6Hz, 1H), 8.53–8.43 (m, 1H), 8.17–8.13 (m, 0.3H), 8.03–7.97 (m, 1H), 7.94–7.87 (m, 1H), 7.85–7.77 (m, 1H), 7.62–7.46 (m, 4H), 7.43–7.33 (m , 2H), 7.09-6.99 (m, 1H), 6.65-6.48 (m, 1H), 5.66-5.53 (m, 1H) 4.74-4.63 (m, 1H ), 4.50-4.39(m, 1H), 4.36-4.27(m, 1H), 4.26-4.16(m, 1H), 4.09-3.98(m, 1H), 3 .88-3.72(m, 2H), 3.70-3.53(m, 2H), 3.27-3.19(m, 3H), 3.18-2.96(m, 2H), 2.9 6-2.72(m, 7H), 2.70-2.59(m, 2H), 2.54-2.33(m, 2H), 2.22-2.05(m, 2H), 1.95- 1.81 (m, 1H), 1.74-1.55 (m, 2H), 1.46 (d, J=6.0Hz, 3H), 1.10-1.02 (m, 3H), 0.95 (dd, J=6.4, 3.2Hz, 1.5H), 0.89-0.85 (m, 1.5H), 0.85-0.71 (m, 6H), 0.64 (s, 3H).

Compound 13G was prepared according to the following scheme:

Step 1: Preparation of methyl 3-formylcyclobutanecarboxylate (compound 13B)

Trifluoroacetic acid (5.1 g, 44.82 mmol) was added to a solution of methyl 3-(methoxymethylene)cyclobutanecarboxylate (compound 3c, 3.5 g, 22.41 mmol) in DCM (100 mL) and water (10 mL). After stirring at 20 °C for 3 hours, the reaction mixture was diluted with _H₂O (60 mL) and extracted three times with DCM (50 mL). The organic phase was washed with brine (80 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give methyl 3-formylcyclobutanecarboxylate (compound 13B, 3 g) as a colorless oil. ^¹H NMR (400 MHz, CDCl₃) δppm 9.82–9.68 (m, 1H), 3.72–3.69 (m, 3H), 3.26–3.07 (m, 2H), 2.56–2.39 (m, 4H).

Step 2: Preparation of methyl 3-ethynylcyclobutanecarboxylate (compound 13C)

Dimethyl dimethyl (1-diazo-2-oxopropyl)phosphonate (7.2 g, 37.46 mmol) and potassium carbonate (6.47 g, 46.83 mmol) were added to a solution of methyl 3-oxocyclobutanecarboxylate (compound 13B, 3.0 g, 23.41 mmol) in methanol (30 mL) cooled to 0 °C. The resulting mixture was stirred at 0 °C for 1 h, then heated to 20 °C for 3 h. The reaction mixture was quenched with water (80 mL) and then extracted twice with PE (60 mL). The combined organic phases were washed with brine (80 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give a residue. The residue was purified by silica gel chromatography, eluting with PE, to give methyl 3-ethynylcyclobutanecarboxylate (compound 13C, 1.5 g) as a colorless oil. ¹ H NMR (400MHz, CDCl3) δppm 3.71-3.66 (m, 3H), 3.34-3.10 (m, 1H), 3.08-2.89 (m, 1H), 2.63-2.49 (m, 2H), 2.47-2.32 (m, 2H), 2.23-2.16 (m, 1H).

Step 3: Preparation of methyl 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylate (compound 13D)

TEA (732 mg, 7.24 mmol), methyl 3-ethynylcyclobutanecarboxylate (compound 13C, 500 mg, 3.62 mmol), tetrakis(triphenylphosphine)palladium(O) (418 mg, 0.360 mmol), and copper iodide(I) (68 mg, 0.36 mmol) were added to a solution of methyl 3-ethynylcyclobutanecarboxylate (500 mg, 3.62 mmol), tetrakis(triphenylphosphine)palladium(O) (418 mg, 0.360 mmol), and copper iodide(I) (68 mg, 0.36 mmol). The reaction mixture was degassed, purged three times with nitrogen, and stirred at 50 °C for 1 hour. After the reaction was complete, the mixture was diluted with water (60 mL) and extracted twice with EtOAc (40 mL). The combined organic phases were washed with brine (60 mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give the residue. The residue was purified by silica gel chromatography (EtOAc = 0-50% in PE) to give methyl 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylate (compound 13D, 500 mg) as a yellow oil. ^¹H NMR (400 MHz, CDCl₃) δppm 8.55 (d, J = 4.4 Hz, 1H), 7.66–7.59 (m, 1H), 7.38 (d, J = 7.6 Hz, 1H), 7.23–7.17 (m, 1H), 3.72–3.69 (m, 3H), 3.48–3.26 (m, 1H), 3.25–3.02 (m, 1H), 2.71–2.47 (m, 4H).

Step 4: Preparation of 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylic acid (compound 13E)

Methyl 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylic acid (compound 13D, 500 mg, 2.32 mmol) was slowly added to a solution of THF (2.5 mL) and water (2.5 mL). The reaction mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the reaction mixture was concentrated under vacuum to remove THF and then acidified to pH 5 with 1 M HCl aqueous solution. The mixture was extracted with EtOAc (80 mL × 2). The combined organic phases were washed with brine (15 mL), dried over _Na₂SO₄ , _filtered , and concentrated under vacuum to give 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylic acid (compound 13E, 500 mg) as a yellow oil. ¹ H NMR (400MHz, CDCl3) δppm 8.58 (d, J=4.4Hz, 1H), 7.68 (t, J=7.6Hz, 1H), 7.44-7.37 (m, 1H), 7.25 (d, J=5.6Hz, 1H), 3.54-3.34 (m, 1H), 3.2-3.10 (m, 1H), 2.71-2.61 (m, 4H).

Step 5: Preparation of methyl (2S)-3-methyl-2-[methyl-[3-[2-(2-pyridyl)ethynyl]cyclobutanecarbonyl]amino]butyrate (compound 13F)

DIEA (192.7 mg, 1.49 mmol) and HATU (207 mg, 0.55 mmol) were added to a solution of 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylic acid (compound 13E, 100 mg, 0.500 mmol) in DMF (3 mL). After stirring at 20 °C for 15 min, methyl (2S)-3-methyl-2-(methylamino)butyrate hydrochloride (99 mg, 0.55 mmol) was added. The reaction mixture was stirred at 20 °C for 12 h. After the reaction was complete, the mixture was purified by reversed-phase chromatography to give methyl (2S)-3-methyl-2-[methyl-[3-[2-(2-pyridyl)ethynyl]cyclobutanecarbonyl]amino]butyrate (compound 13F, 160 mg) as a yellow oil. MS: Calculated value 329.2 (MH ⁺ ); Measured value 329.3 (MH ⁺ )

Step 6: Preparation of (2S)-3-methyl-2-[methyl-[3-[2-(2-pyridyl)ethynyl]cyclobutanecarbonyl]amino]butyric acid (compound 13G)

Lithium hydroxide monohydrate (19 mg, 0.49 mmol) was added to a solution of methyl (2S)-3-methyl-2-[methyl-[3-[2-(2-pyridyl)ethynyl]cyclobutanecarbonyl]amino]butyrate (compound 13F, 80 mg, 0.24 mmol) in water (2 mL) and THF (2 mL). The reaction mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the mixture was concentrated under vacuum to remove THF, and the aqueous solution was acidified to pH 5 with 1N HCl. The resulting mixture was extracted twice with EtOAc (15 mL), and the combined organic phases were washed with brine ( ₂₀ mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give (2S)-3-methyl-2-[methyl-[3-[2-(2-pyridyl)ethynyl]cyclobutanecarbonyl]amino]butyric acid (compound 13G, 60 mg) as a yellow oil. MS calculated value 315.2 (MH ⁺ ); measured value 315.2 (MH ⁺ ).

Example 14

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-ethynyl-N-methyl-azacyclobutane-1-carboxamide

The title compound was prepared in a manner similar to that of Example 13, using 3-ethynyltetrahydroacryl instead of 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylic acid (compound 13E). Example 14 (4 mg) was obtained as a white solid. MS calculated value 860.5 (MH ⁺ ); measured value 860.4 (MH ⁺ ). ^1H NMR (400 MHz, MeOD) δppm 8.74-8.73 (m, 1H), 8.0 (d, J=13.2Hz, 1H), 7.90 (d, J=8Hz, 1H), 7.60-7.49 (m, 3H), 7.36 (d, J=7.2Hz, 1H), 7.04 (s, 1H), 6.5 4(s, 1H), 5.66-5.63(m, 1H), 4.32-4.27(m, 1H), 4.21-4.18(m, 1H), 4.07-3.95(m, 2H), 3.77(s, 1H), 3.66-3.42(m, 2H), 3.0 0-2.94(m, 4H), 2.76(s, 3H), 2.69(s, 3H), 2.37-2.33(t, J=7.2Hz, 1H), 2.15-2.10(m, 2H), 2.04-2.02(m, 1H), 1.94-1.90(m , 1H), 1.69-1.57(m, 4H), 1.47-1.45(d, 6.0Hz, 3H), 1.29-1.28(m, 4H), 1.07-1.03(m, 3H), 0.93-0.83(m, 9H), 0.64(s, 3H).

Example 15

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-4-ethynyl-N-methyl-piperidine-1-carboxamide

The title compound was prepared in a manner similar to that of Example 13 by using 4-ethynylpiperidine instead of 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylic acid (compound 13E). Example 15 (5 mg) was obtained as a white solid. MS calculated value 888.5 (MH ⁺ ); measured value 888.6 (MH ⁺ ). ^1H NMR (400 MHz, DMSO- _d6 ) δppm 9.30(s, 1H), 8.78-8.72(m, 1H), 8.48(s, 1H), 8.14-8.06(m, 1H), 7.91(s , 1H), 7.86-7.78(m, 1H), 7.63-7.57(m, 1H), 7.56-7.48(m, 2H), 7.32-7.2 3(m, 1H), 7.06-6.98(m, 1H), 6.40-6.32(m, 1H), 5.51-5.41(m, 1H), 5.38 -5.29(m,1H),4.33-4.16(m,4H),4.09-3.96(m,2H),3.89-3.78(m,3H),3 .68-3.62(m, 2H), 3.63-3.52(m, 2H), 3.12(s, 3H), 3.07-2.89(m, 3H), 2. 87-2.74(m,2H),2.69(s,1H),2.61(s,2H),2.10-1.92(m,2H),1.87-1.74 (m, 3H), 1.69-1.56 (m, 2H), 1.55-1.46 (m, 2H), 1.42-1.35 (m, 3H), 1.00-0 .92 (m, 3H), 0.83 (d, J=6.4Hz, 3H), 0.80-0.74 (m, 6H), 0.58-0.46 (m, 3H).

Example 16

N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-4-ethynyl-4-fluoro-N-methyl-piperidine-1-carboxamide

The title compound was prepared in a manner similar to that of Example 13 by using 4-ethynyl-4-fluoropiperidine instead of 3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxylic acid (compound 13E). Example 16 (6 mg) was obtained as a white solid. MS calculated value 906.5 (MH ⁺ ); measured value 906.3 (MH ⁺ ). ^1H NMR (400 MHz, DMSO-d6) δppm 9.33 (s, 1H), 8.79-8.73 (m, 1H), 8.45 (s, 1H), 8.14-8.05 (m, 1H), 7.97-7.92 (m, 1H), 7.86-7.79 (m, 1H), 7.65-7 .49(m, 3H), 7.21(s, 1H), 7.06-6.95(m, 1H), 6.40-6.31(m, 1H), 5.55-5.25(m, 2H), 4.34-4.18(m, 4H), 4.08-3. 77(m, 9H), 3.71-3.61(m, 4H), 3.13-3.05(m, 3H), 3.02-2.71(m, 1H), 2.63-2.60(m, 3H), 2.12-1.91(m, 6H), 1.4 2-1.36 (m, 3H), 1.26-1.13 (m, 2H), 1.00-0.91 (m, 3H), 0.87-0.80 (m, 4H), 0.80-0.73 (m, 6H), 0.60-0.43 (m, 3H).

Example 17

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

By using (2R)-2-chloro-2-fluoroacetic acid and (7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate H) instead of (2S)-2-chloro-2-fluoro The title compound was prepared in a manner similar to that of Example 1 using acetic acid (1 g of compound) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C). Example 17 (47 mg) was obtained as a white solid. MS calculated value 935.4 (MH ⁺ ); measured value 935.6 (MH ⁺ ). ¹ H NMR (400MHz, MeOD) δppm8.77-8.68 (m, 1H), 8.60-8.56 (m, 1H), 7.89-7.80 (m, 1H), 7.76-7.66 (m, 1H), 7.59-7.43 (m, 3H), 6.95-6.73 (m, 1H), 5.85-5.67 (m, 1H), 4.84-4.77 (m, 2H), 4.47-4.34 (m, 2H), 4.33-4.06 (m, 3H), 3.98-3.66 (m, 6H), 3.63-3.39 (m, 2H), 3.38-3. 33(m, 3H), 3.30-3.20(m, 1H), 3.14-2.99(m, 4H), 2.86-2.74(m, 1H), 2.64-2.53(m, 1H), 2.38-2.02(m, 4H), 1.99-1.90(m, 1H), 1.87- 1.72 (m, 1H), 1.69-1.54 (m, 1H), 1.45 (d, J=6.0Hz, 3H), 1.02-0.96 (m, 3H), 0.95-0.92 (m, 5H), 0.91-0.85 (m, 3H), 0.57-0.40 (m, 3H).

Example 18

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

By using (2R)-2-chloro-2-fluoro-acetic acid and (7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate F) and (2S)-2-(2-(tert-butylcarbonyl)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl)-3-methyl The title compound was prepared in a manner similar to that of Example 1 by replacing (2S)-2-chloro-2-fluoro-acetic acid (compound 1 g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C) and BOC-N-ME-VAL-OH (compound 1a). Example 18 (25.4 mg) was obtained as a white solid. MS calculated value 1059.5 (MH ⁺ ); measured value 1059.4 (MH ⁺ ). ^¹H NMR (400MHz, methanol- _d4) )δ=8.59-8.55(m, 1H), 8.41(d, J=2.4Hz, 1H), 7.74-7.68(m, 1H), 7.62-7.54(m, 1H), 7.51-7.46(m, 1H), 7.35-7.31(m, 1H), 6.9 4-6.74 (m, 1H), 5.78-5.66 (m, 1H), 4.59-4.39 (m, 2H), 4.32-4.20 (m, 4H), 4.11-3.97 (m, 1H), 3.94-3.86 (m, 1H), 3.81-3.67 (m, 4 H), 3.64-3.52(m, 2H), 3.49-3.41(m, 3H), 3.39-3.35(m, 4H), 3.13-3.07(m, 1H), 2.81-2.59(m, 7H), 2.46-2.38(m, 4H), 2.33-2. 20 (m, 2H), 1.98-1.76 (m, 8H), 1.67-1.61 (m, 1H), 1.41 (d, J=6.0Hz, 3H), 1.02-0.95 (m, 9H), 0.91-0.87 (m, 3H), 0.47 (s, 3H)ppm.

Compound 18l was prepared according to the following scheme:

Step 1: Preparation of 1-(tert-butyl)-3-methyl-3-(but-3-en-1-yl)pyrrolidine-1,3-dicarboxylate (compound 18c)

At -70 °C under a nitrogen atmosphere, lithium diisopropylimide (59.9 mL, 119.95 mmol) was added to a solution of 1-(tert-butyl)-3-methylpyrrolidine-1,3-dicarboxylic acid (compound 18a, 25.0 g, 109.04 mmol) in THF (300 mL). After stirring for 0.5 h, 4-bromo-1-butene (compound 18b, 16.1 g, 119.95 mmol) was added. The mixture was stirred at 20 °C for 2.5 h. The mixture was quenched by adding saturated _NH₄Cl (100 mL) and extracted twice with EtOAc (150 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified and concentrated by silica gel column chromatography (EtOAc in PE = 0%–5%) to give a yellow oily substance, 1-(tert-butyl)-3-methyl-3-(but-3-en-1-yl)pyrrolidine-1,3-dicarboxylic acid (compound 18c, 13.2 ^g ). NMR (400MHz, methanol-d4) δ＝5.83-5.73(m, 1H), 5.05-4.93(m, 2H), 3.84(d, J＝10.8Hz, 1H), 3.71(s, 3H), 3.46-3.37(m, 1H), 3.29-3.11(m, 2H), 2.40-2.32(m, 1H), 2.01-1.91(m, 2H), 1.90-1.74(m, 3H), 1.46(d, J＝3.2Hz, 9H)ppm.

Step 2: Preparation of methyl 3-(but-3-en-1-yl)pyrrolidine-3-carboxylate (compound 18d)

HCl/1,4-dioxane (50.0 mL, 4 M) was added to a solution of 1-(tert-butyl)-3-methyl-3-(but-3-en-1-yl)pyrrolidine-1,3-dicarboxylic acid (compound 18c, 13.2 g, 46.58 mmol) in 1,4-dioxane (50 mL). The mixture was stirred at 20 °C for 0.5 h. The mixture was concentrated under reduced pressure to give methyl 3-(but-3-en-1-yl)pyrrolidine-3-carboxylic acid hydrochloride (compound 18d, 10.2 g) as a yellow oil.

Step 3: Preparation of methyl 3-(but-3-en-1-yl)-1-triphenylmethylpyrrolidone-3-carboxylate (compound 18e)

Triphenylmethyl chloride (3.81 g, 13.65 mmol) and potassium carbonate (4.72 g, 34.14 mmol) were added to a solution of methyl 3-(but-3-en-1-yl)pyrrolidine-3-carboxylate hydrochloride (compound 18d, 3.0 g, 13.65 mmol) in ACN (70 mL). The mixture was stirred at 20 °C for 12 h. After the reaction was complete, the mixture was added to water (100 mL) and extracted three times with EtOAc (30 mL). The combined organic phases were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 0%–5%) and concentrated to give methyl 3-(but-3-en-1-yl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 18e, 1.3 g) as a yellow oil. ¹ H NMR (400MHz, CDCl3) δ = 7.48-7.45 (m, 6H), 7.26-7.23 (m, 6H), 7.15 (t, J = 7.2Hz, 3H), 5.78-5.68 (m, 1H), 5.00-4.89 (m, 2H), 3.74 (s, 3H), 3.00 (d, J =9.6Hz, 1H), 2.61-2.55(m, 1H), 2.43-2.35(m, 1H), 2.20-2.14(m, 1H), 2 .03 (s, 1H), 1.98-1.83 (m, 2H), 1.82-1.61 (m, 2H), 1.52-1.45 (m, 1H)ppm.

Step 4: Preparation of methyl 3-(3-oxopropyl)-1-triphenylmethylpyrrolidone-3-carboxylate (compound 18f)

Potassium osmium tetroxide (VI) (5 mg, 0.02 mmol) was added to a solution of methyl 3-(but-3-en-1-yl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 18e, 660 mg, 1.55 mmol) in THF (20 mL) and water ( ₁₀ mL), followed by sodium periodate (663 mg, 3.1 mmol). The mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the mixture was quenched by adding a saturated aqueous solution of NH₄Cl and extracted twice with EtOAc (50 mL). The organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified and concentrated by silica gel column chromatography (EtOAc in PE = 0%–5%) to give methyl 3-(3-oxopropyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 18f, 300 mg) as a yellow oil. ¹ H NMR (400MHz, CDCl ₃ )δ=9.75-9.69 (m, 1H), 7.52-7.48 (m, 6H), 7.29 (t, J=4.0Hz, 6H), 7.19 (t, J=7.2Hz, 3H), 3.77 (s, 3H), 3.05 (d, J= 9.2Hz, 1H), 2.67-2.57(m, 1H), 2.47-2.31(m, 3H), 2.29-2.21(m, 1H), 2.04-1.87(m, 2H), 1.62-1.43(m, 2H)ppm.

Step 5: Preparation of methyl 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 18h)

Sodium cyanoborohydride (88.9 mg, 1.42 mmol) was added to a stirred solution of methyl 3-(3-oxopropyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 18f, 550.0 mg, 1.29 mmol), H-VAL-OTBU HCl salt (compound 18 g, 296.76 mg, 1.42 mmol), and zinc chloride (192.8 mg, 1.42 mmol) in methanol (6 mL). The mixture was stirred at 25 °C for 2 h. After the reaction was complete, water (30 mL) was added to the reaction mixture. The resulting mixture was concentrated under reduced pressure to remove MeOH. The resulting suspension was diluted with water (40 mL) and extracted twice with EtOAc (30 mL). The organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (EtOAc in PE = 0%–10%) to give a yellow oily compound, methyl 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobut-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 18h, 510 mg). ^¹H NMR (400 MHz, CDCl₃ ₎ )δ=7.47(d, J=8.0Hz, 6H), 7.28-7.27(m, 1H), 7.27-7.23(m, 5H), 7.18-7.13(m, 3H), 3.78-3. 73 (m, 3H), 3.05 (t, J=8.4Hz, 1H), 2.79 (dd, J=2.4, 6.0Hz, 1H), 2.67-2.59 (m, 1H), 2.55-2.49 (m, 1H), 2.43-2.31 (m, 2H), 2.14-2.09 (m, 1H), 1.99-1.94 (m, 1H), 1.88-1.82 (m, 1H), 1.68-1 .64(m, 3H), 1.60-1.55(m, 2H), 1.48-1.45(m, 9H), 1.37-1.33(m, 1H), 0.94-0.90(m, 6H)ppm.

Step 6: Preparation of 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylic acid (compound 18i)

Lithium hydroxide monohydrate (336 mg, 8 mmol) was added to a solution of methyl 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 18h, 510 mg, 0.8 mmol) in methanol (6 mL), water (0.6 mL), and THF (0.6 mL). The mixture was stirred at 60 °C for 12 h. After the reaction was complete, the mixture was diluted with water (20 mL), neutralized with 1 M HCl aqueous solution, and extracted twice with EtOAc (20 mL). The organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobut-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylic acid (compound 18i, 440 mg), which was a white gel. ¹ H NMR (400MHz, CDCl ₃ )δ=7.46(d, J=8.0Hz, 6H), 7.26-7.19(m, 6H), 7.16-7.10(m, 3H), 3.10-2.86(m, 4H), 2.54-2.45(m, 2H), 2.37-2.22(m, 2H) ), 2.18-2.10(m, 1H), 2.04-1.97(m, 1H), 1.68-1.54(m, 2H), 1.49-1.44(m, 9H), 1.41-1.35(m, 2H), 0.99-0.89(m, 6H)ppm.

Step 7: Preparation of tert-butyl (2S)-3-methyl-2-(6-oxo-2-triphenylmethyl-2,7-diazaspiro[4.5]decane-7-yl)butyrate (compound 18j)

At 0 °C, DIEA (0.67 mL, 3.85 mmol) and COMU (4-{{[(1-cyano-2-ethoxy-2-oxoethylidene)amino]oxaloyl](dimethylamino)methylene]-hexafluorophosphate, CAS1075198-30-9) (594 mg, 1.39 mmol) were added to a solution of 3-(3-((S)-1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl)amino]oxaloyl](dimethylamino)methylene]-hexafluorophosphate, CAS1075198-30-9) in DMF (5 mL). The mixture was stirred at 20 °C for 1 hour. After the reaction was complete, the mixture was diluted with water (20 mL) and extracted twice with EtOAc (20 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (EtOAc in PE 0%–10%) to give tert-butyl (2S)-3-methyl-2-(6-oxo-2-triphenylmethyl-2,7-diazaspiro[4.5]decane-7-yl)butyrate (compound 18j, 420 mg), a yellow oil. MS calculated value 553.3 (MH ⁺ ); measured value 553.2 (MH ⁺ ).

Step 8: Preparation of (2S)-3-methyl-2-(6-oxo-2,7-diazaspiro[4.5]decane-7-yl)butyric acid (compound 18k)

TFA (3.0 mL) was added to a solution of (2S)-3-methyl-2-(6-oxo-2-triphenylmethyl-2,7-diazaspiro[4.5]decane-7-yl)butyrate (compound 18j, 550 mg, 1 mmol) in DCM (3 mL). The mixture was stirred at 20 °C for 12 h. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain a residue, which was dissolved in water (10 mL) and extracted twice with EtOAc (20 mL). The aqueous phase was concentrated under vacuum to give (2S)-3-methyl-2-(6-oxo-2,7-diazaspiro[4.5]decane-7-yl)butyrate (compound 18k, 365 mg) as a colorless oil in the form of a TFA salt. MS calculated value 255.2 (MH ⁺ ); measured value 255.2 (MH ⁺ ).

Step 9: Preparation of (2S)-2-(2-(tert-butoxycarbonyl)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl)-3-methylbutyric acid (compound 181).

Sodium carbonate (210 mg, 1.98 mmol) and di-tert-butyl dicarbonate (259 mg, 1.19 mmol) were added to a solution of (2S)-3-methyl-2-(6-oxo-2,7-diazaspiro[4.5]decane-7-yl)butyric acid (compound 18k, 365 mg, 0.99 mmol) in THF (10 mL) and water (10 mL). The mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the mixture was neutralized with 1 M HCl aqueous solution and extracted twice with EtOAc (20 mL). The organic phase was washed with brine, dried over sodium sulfate, and concentrated under vacuum to give (2S)-2-(2-(tert-butoxycarbonyl)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl)-3-methylbutyric acid (compound 18k, 350 mg) as a white solid.

Example 19

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decanev7-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

By using (2R)-2-chloro-2-fluoro-acetic acid and (7S,13S)-7-amino-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate I) and (2S)-2-(2-(tert-butylcarbonyl)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl The title compound was prepared in a manner similar to that of Example 1 by replacing (2S)-2-chloro-2-fluoro-acetic acid (compound 1g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C) and BOC-N-ME-VAL-OH (compound 1a). Example 19 (54.4 mg) was obtained as a white solid. MS calculated value 1113.5 (MH ⁺ ); measured value 1113.3 (MH ⁺ ). ¹ H NMR (400MHz, methanol-d4) δ = 8.61-8.57 (m, 1H), 8.44 (d, J = 2.4Hz, 1H), 7.81-7.7 4(m, 1H), 7.68-7.57(m, 2H), 7.34(s, 1H), 6.95-6.72(m, 1H), 5.72-5.62(m , 1H), 5.20-5.10 (m, 1H), 4.99-4.92 (m, 1H), 4.84-4.77 (m, 2H), 4.70-4.56 (m, 1H), 4.45-4.38 (m, 1H), 4.32-4.25 (m, 1H), 4.21-4.15 (m, 1H), 4.11-3. 79(m, 2H), 3.77-3.65(m, 3H), 3.59-3.53(m, 1H), 3.51-3.33(m, 10H), 3.21 -3.15(m, 1H), 2.5-2.71(m, 5H), 2.61-2.55(m, 1H), 2.45(s, 3H), 2.33-2.2 0 (m, 2H), 2.02-1.78 (m, 7H), 1.68-1.59 (m, 1H), 1.43 (d, J=6.0Hz, 3H), 1.0 2(d, J=6.4Hz, 1H), 1.00-0.97(m, 4H), 0.93-0.79(m, 4H), 0.42(s, 3H)ppm.

Example 20

1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidin-4-carboxamide

By using (2R)-2-chloro-2-fluoro-acetic acid and (7S,13S)-7-amino-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate I) and 1-BOC-4-fluoro-4-piperidinic acid instead of (2S). The title compound was prepared in a manner similar to that of Example 1 using 2-chloro-2-fluoro-acetic acid (compound 1 g), (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C), and (S)-1-BOC-pyrrolidine-3-carboxylic acid (compound 1 d). Example 20 (94.41 mg) was obtained as a yellow solid. MS calculated value 1119.5 (MH ⁺ ); measured value 1119.5 (MH ⁺ ). ¹ H NMR (400MHz, methanol-d4) δ = 8.64-8.58 (m, 1H), 8.52-8.49 (m, 1H), 7.85-7.75 (m, 1H), 7.65-7.60 (m, 2H), 7.54-7.50 (m, 1H), 7.18-6.96 (m, 1H), 5 .82-5.61(m, 1H), 5.33-5.10(m, 2H), 4.54-4.36(m, 2H), 4.33-4.20(m , 2H), 4.16-3.87(m, 4H), 3.83-3.66(m, 1H), 3.85-3.62(m, 4H), 3.60-3 .43(m, 4H), 3.27-3.19(m, 6H), 3.10-3.05(m, 1H), 3.03-2.99(m, 3H), 2.89-2.76(m, 1H), 2.63-2.50(m, 1H), 2.39-2.11(m, 6H), 2.05-1.93(m , 1H), 1.91-1.76 (m, 1H), 1.49-1.38 (m, 4H), 1.35-1.28 (m, 1H), 1.15- 1.07 (m, 1H), 1.05-0.96 (m, 7H), 0.94-0.85 (m, 3H), 0.51-0.34 (m, 3H).

Example 21

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

By using (2R)-2-chloro-2-fluoroacetic acid and (7S,13S)-7-amino-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate I) instead The title compound was prepared in a manner similar to that of Example 1 using (2S)-2-chloro-2-fluoro-acetic acid (compound 1 g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C). Example 21 (85.7 mg) was obtained as a white solid. MS calculated value 1087.5 (MH ⁺ ); measured value 1087.3 (MH ⁺ ). ¹ H NMR (400MHz, MeOD) δ = 8.62-8.58 (m, 1H), 8.52-8.48 (m, 1H), 7.82-7.74 (m, 1H), 7.66-7.57 (m, 2H), 7.53 (br d, J=3.4Hz, 1H), 6.94-6.76 (m, 1H), 5.79-5.61 (m, 1H), 5.28-5.12 (m, 1H), 4.97-4.88 (m, 1H), 4.84-4.78 (m, 1H), 4. 4-4.38(m, 1H), 4.32-4.21(m, 2H), 4.12-3.93(m, 2H), 3.90-3.56(m, 9H), 3.54-3.33(m, 6H), 3.30-3.16(m, 4H), 3.1 5-3.09 (m, 3H), 3.01-2.97 (m, 3H), 2.86-2.75 (m, 1H), 2.61-2.50 (m, 1H), 2.46-2.16 (m, 4H), 2.01-1.92 (m, 1H), 1.8 8-1.75 (m, 1H), 1.70-1.57 (m, 1H), 1.45 (d, J=6.0Hz, 3H), 1.10-0.96 (m, 6H), 0.95-0.85 (m, 3H), 0.50-0.38 (m, 3H).

Example 22

(3S)-1-[(2R)-2-chloro-2-fluoroacetyl]-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

By using (2R)-2-chloro-2-fluoroacetic acid and (7S,13S)-7-amino-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (compound 22a) instead of (2S)-2 The title compound was prepared in a manner similar to that of Example 1 using chloro-2-fluoro-acetic acid (compound 1 g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C). Example 22 (66.5 mg) was obtained as a white solid. MS calculated value 989.3 (MH ⁺ ); measured value 989.4 (MH ⁺ ). ¹ H NMR (400MHz, MeOD) δ=8.80 (dd, J=4.8, 1.2, 1H), 8.74-8.47 (m, 2H), 8.02 (d, J=7 .6Hz, 1H), 7.85-7.74(m, 1H), 7.73-7.67(m, 1H), 7.66-7.53(m, 2H), 6.95-6.74 (m, 1H), 5.88-5.60 (m, 1H), 5.31-5.15 (m, 1H), 4.83-4.78 (m, 1H), 4.47-4.32 (m , 2H), 4.31-4.20(m, 1H), 4.14-3.86(m, 1H), 3.85-3.65(m, 5H), 3.63-3.52(m, 1 H), 3.51-3.42(m, 1H), 3.42-3.36(m, 3H), 3.30-3.23(m, 1H), 3.23-3.14(m, 1H) , 3.13-2.99(m, 3H), 2.90-2.73(m, 1H), 2.67-2.50(m, 1H), 2.42-2.03(m, 4H), 2 .00-1.90(m, 1H), 1.89-1.72(m, 1H), 1.70-1.54(m, 1H), 1.47(d, J=6.4Hz, 3H), 1.18-0.99(m,3H), 0.98-0.94(m,3H), 0.94-0.84(m,3H), 0.51-0.35(m,3H)ppm.

Compound 22a was prepared in a manner similar to that used in the preparation of intermediate H by using _CF3CH2OTf instead of _iodoethane .

Examples 23 and 24

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl- 8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 23)

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl- 8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 24)

By using (2R)-2-chloro-2-fluoroacetic acid and (7S,13S)-7-amino-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate I) and compounds 23g1/23g2 instead of (2S The title compound was prepared in a manner similar to that described in Example 1 using 2-chloro-2-fluoro-acetic acid (compound 1 g), (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C) and BOC-N-ME-VAL-OH (compound 1a).

Example 23 (21.1 mg) was obtained as a white solid. MS calculated value 1099.5 (MH ⁺ ); measured value 1099.4 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ=8.59-8.55 (m, 1H), 8.41 (d, J=2.4Hz, 1H), 7.74-7.68 (m, 1H), 7.63 (d, J=4.4Hz, 1H), 7.58 (d, J=8.8Hz, 1H), 7.51-7.46 (m, 1H), 7.35-7 .31 (m, 1H), 6.94-6.74 (m, 1H), 5.78-5.66 (m, 1H), 4.59-4.39 (m, 2H), 4.41-4.25 (m, 3H), 4.20-4.14 (m, 1H), 3.79-3.61 (m, 6H), 3.55-3.4 0(m, 5H), 3.34-3.34(m, 3H), 3.21-3.12(m, 2H), 2.81-2.79(m, 1H), 2.68-2.58(m, 5H), 2.36(s, 3H), 2.28-2.07(m, 6H), 2.05-1.95(m, 2H) , 1.90-1.77 (m, 1H), 1.73-1.58 (m, 1H), 1.43 (d, J = 6.0Hz, 3H), 1.02 (d, J = 6.4Hz, 3H), 0.99 (s, 3H), 0.90 (d, J = 6.4Hz, 3H), 0.43 (s, 3H) ppm.

Example 24 (11.5 mg) was obtained as a white solid. MS calculated value 1099.5 (MH ⁺ ); measured value 1099.7 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4 ) δ＝8.60 (s, 1H), 8.43 (d, J＝2.8 Hz, 1H), 7.84–7.72 (m, 1H), 7.68 (s, 1H), 7.59 (d, J＝10.0 Hz, 1H), 7.33 (s, 1H), 6.97–6.70 (m, 1H), 5.75–5.65 (m, 1H), 5.16 (br dd, J＝8.4, 16.0 Hz, 1H), 4.43–4.24 (m, 3H), 4.18 (br dd, J＝8.4, 16.0 Hz, 1 ... d, J=6.0Hz, 1H), 4.01-3.89 (m, 1H), 3.88-3.66 (m, 5H), 3.63-3.56 (m, 2H), 3.52-3.41 (m, 3H), 3.39-3.35 (m, 4 H), 3.34-3.31(m, 3H), 3.30-3.25(m, 1H), 3.22-3.13(m, 1H), 2.88-2.77(m, 1H), 2.75-2.60(m, 3H), 2.62-2.5 4(m, 1H), 2.41(s, 3H), 2.35-2.19(m, 3H), 2.17-2.04(m, 3H), 2.02-1.95(m, 1H), 1.84(brd, J=13.2Hz, 1H), 1. 70-1.57 (m, 1H), 1.43 (d, J=6.0Hz, 3H), 1.03-0.97 (m, 6H), 0.91 (dd, J=3.6, 6.4Hz, 3H), 0.46-0.38 (m, 3H)ppm.

Compounds 23g1 and 23g2 were prepared according to the following scheme:

Step 1: Preparation of 1-(tert-butyl)-3-methyl-3-allylpyrrolidine-1,3-dicarboxylic acid (compound 23b).

At -70 °C under a nitrogen atmosphere, LDA (12 mL, 24 mmol) was added dropwise to a solution of 1-(tert-butyl)-3-methylpyrrolidine-1,3-dicarboxylic acid (compound 23a, 5.0 g, 21.8 mmol) in THF (60 mL). After stirring for 0.5 h, propenyl bromide (2.9 g, 23.99 mmol) was slowly added. After the reaction was complete, the mixture was poured into a saturated aqueous solution of _NH₄Cl (100 mL) and extracted twice with EtOAc (70 mL). The combined organic phases were washed with brine (70 mL), dried over _Na₂SO₄ , filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column _{chromatography} (EtOAc in PE = 1%–20%) to give 1-(tert-butyl)-3-methyl-3-allylpyrrolidine-1,3-dicarboxylic acid (compound 23b, 2.91 g) as a colorless oil.

Step 2: Preparation of 1-(tert-butyl)-3-methyl-3-(2-oxoethyl)pyrrolidine-1,3-dicarboxylic acid (compound 23c)

At 0 °C, 2,6-dimethylpyridine (1.8 mL, 15.6 mmol) and KOsO4 (0.14 g, 0.39 mmol) were added in portions to a mixture of 1-(tert-butyl)-3-methyl- ₃ -allylpyrrolidine-1,3-dicarboxylic acid (compound 23b, 2.1 g, 7.8 mmol) in 1,4-dioxane (60 mL) and water (6 mL). After stirring at 0 °C for 15 min, sodium periodate (6.67 g, 31.19 mmol) was added in portions at 0 °C. The resulting mixture was warmed to 20 °C and stirred for another 6 h. After the reaction was complete, the mixture was quenched with saturated _{Na2S2O3 aqueous} solution (100 mL) and extracted three times with EtOAc (50 mL). The combined organic phases were washed with brine (50 mL), dried with _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to give 1-(tert-butyl)-3-methyl-3-(2-oxoethyl)pyrrolidine-1,3-dicarboxylic acid (compound 23c, 2.1 g), which was a yellow oil and was used directly in the next step.

Step 3: Preparation of 1-(tert-butyl)3-methyl-3-(2-(((S)-1-(benzyloxy)-3-methyl-1-oxobut-2-yl)amino)ethyl)pyrrolidine-1,3-dicarboxylic acid (compound 23d).

At 0 °C, zinc chloride (1.05 g, 7.74 mmol) was added in part one portion to a mixture of 1-(tert-butyl)-3-methyl-3-(2-oxoethyl)pyrrolidine-1,3-dicarboxylic acid (compound 23c, 2.1 g, 7.74 mmol) and (2S)-2-amino-3-methyl-butyrate benzyl ester (1.6 g, 7.74 mmol) in methanol (20 mL). After stirring at 0 °C for 1 h, sodium cyanoborohydride (0.97 g, 15.48 mmol) was added to the mixture at 0 °C. The resulting mixture was stirred at 0 °C for another 2 h. After the reaction was complete, the mixture was added to a saturated aqueous solution of _NH₄Cl (40 mL) at 0 °C and extracted three times with EtOAc (50 mL). The combined organic phases were washed four times with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by silica gel column chromatography (EtOAc in PE = 1%–25%) to give a yellow oily compound, 1-(tert-butyl)-3-methyl-3-(2-(((S)-1-(benzyloxy)-3-methyl-1-oxobut-2-yl)amino)ethyl)pyrrolidine-1,3-dicarboxylic acid (compound 23d, 2.2 g). MS calculated value 463.3 (MH ⁺ ); measured value 463.2 (MH ⁺ ).

Step 4: Preparation of tert-butyl 7-((S)-1-(benzyloxy)-3-methyl-1-oxobut-2-yl)-6-oxo-2,7-diaminospiro[4.4]nonane-2-carboxylate (compounds 23e and 23f).

A mixture of 1-(tert-butyl)-3-methyl-3-(2-((((S)-1-(benzyloxy)-3-methyl-1-oxobut-2-yl)amino)ethyl)pyrrolidine-1,3-dicarboxylic acid (compound 23d, 2.1 g, 4.54 mmol) in toluene (20 mL) was mixed with DIEA (7.9 mL, 45.4 mmol) and DMAP (0.55 g, 4.54 mmol) in one part. The mixture was heated to 80 °C and stirred for 16 hours. After the reaction was complete, the mixture was poured into water (30 mL) and extracted three times with EtOAc (30 mL). The combined organic phases were washed with brine (30 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to give a residue. The residue was purified by reversed-phase combined rapid chromatography, and the eluent was concentrated under vacuum. The residue was further separated by preparative SFC (0.1% NH3 _{· H2O} -MeOH conditions, B% = 60%, gradient time = 7.5 min) and concentrated under vacuum to give tert-butyl 7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate to give tert-butyl 7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (compound 23e, faster elution, 521 mg; compound 23f, slower elution, 525 mg). MS: calculated value 453.3 (MNa ⁺ ); measured value 453.2 (MNa ⁺ ).

Step 5: Preparation of (S)-2-((R)-7-(tert-butoxycarbonyl)-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl)-3-methylbutyric acid (compound 23g1).

To a solution of (R)-7-((S)-1-(benzyloxy)-3-methyl-1-oxobut-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (compound 23e, 120 mg, 0.28 mmol) in toluene (2 mL), wet palladium (12 mg, based on 10% wt. of activated carbon) was added. The mixture was degassed and purged three times with hydrogen. The mixture was heated to 35 °C and stirred under hydrogen for 3 hours. After the reaction was complete, the solution was filtered through a diatomaceous earth pad and the filtrate was concentrated under vacuum to give (S)-2-((R)-7-(tert-butoxycarbonyl)-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl)-3-methylbutyric acid (compound 23g, 94 mg) as a white solid. MS calculated value 363.2 (MNa ⁺ ); measured value 363.1 (MNa ⁺ ).

Step 6: Preparation of (S)-2-((S)-7-(tert-butoxycarbonyl)-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl)-3-methylbutyric acid (compound 23g2).

To a solution of (S)-7-((S)-1-(benzyloxy)-3-methyl-1-oxobut-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (compound 23f, 120 mg, 0.28 mmol) in toluene (2 mL), wet palladium (12 mg, based on 10% wt. of activated carbon) was added. The mixture was degassed and purged three times with hydrogen. The mixture was heated to 35 °C and stirred under hydrogen for 3 h. After the reaction was complete, the solution was filtered through a diatomaceous earth pad and the filtrate was concentrated under vacuum to give (S)-2-((S)-7-(tert-butoxycarbonyl)-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl)-3-methylbutyric acid (compound 23f, 2.79 mg) as a white solid. MS calculated value 363.2 (MNa ⁺ ); measured value 363.1 (MNa ⁺ ).

Example 25

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

By using (2R)-2-chloro-2-fluoro-acetic acid and (7S,13S)-7-amino-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (compound 25a) instead of (2S The title compound was prepared by means of (8S,14S)-2-chloro-2-fluoro-acetic acid (compound 1 g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate C) in a manner similar to that of Example 1. Example 25 (68.8 mg) was obtained as a yellow solid. MS calculated value 1074.4.4 (MH ⁺ ); measured value 1074.4 (MH ⁺ ) ^. HNMR (400MHz, MeOD) δ = 8.65-8.60 (m, 1H), 8.45-8.38 (m, 1H), 7.85-7.76 (m, 2H), 7.67-7.57 (m, 2H), 6.94-6.74 (m, 1H), 5.80-5.65 (m, 1H), 5.34-5.22 (m, 1H), 4.88-4.71 (m, 2H), 4.46-4.24 (m, 3H), 3.95-3.58 (m, 12H), 3.45- 3.35 (m, 8H), 3.25-3.09 (m, 4H), 2.86-2.76 (m, 1H), 2.64 (dd, J=5.8, 14.4Hz, 1H), 2-37-2.15 (m, 4H), 2.01-1.92 (m, 1H), 1.88-1.74 (m, 1H), 1.67-1.57 (m, 1H), 1.48 (d, J=6.4Hz, 3H), 1.04-0.85 (m, 9H), 0.56-0.47 (m, 3H)ppm.

Compound 25a was prepared in a manner similar to that used for intermediates F and D by replacing iodoethane and 1-Cbz-piperazine (compound D4) with _{CF3CH2OTf and} morpholine.

Example 26

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

By using (2R)-2-chloro-2-fluoro-acetic acid and (7S,13S)-7-amino-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaen-8,14-dione (compound 26a) The title compound was prepared in a manner similar to that of Example 1, replacing (2S)-2-chloro-2-fluoro-acetic acid (compound 1 g) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26), 2,4,6(29), 20,23(27), 24-heptaene-9,15-dione (intermediate C). Example 26 (3.3 mg) was obtained as a white solid. MS calculated value 1101.4 (MH ⁺ ); measured value 1101.1 (MH ⁺ ). ¹ H NMR (400MHz, MeOD) δ = 8.64-8.37 (m, 3H), 7.79-7.71 (m, 2H), 7.60-7.50 (m, 2H), 6.95-6.74 (m, 1H), 5.90-5.70 (m, 1H), 4.47-4.3 2(m, 3H), 4.26-4.09(m, 2H), 3.97-3.76(m, 5H), 3.71-3.55(m, 2H), 3.50-3.43(s, 5H), 3.43-3.35(m, 4H), 3.23-3.14(m, 3H), 3. 13-3.09(m, 3H), 2.92-2.88(m, 4H), 2.86-2.79(m, 1H), 2.73-2.65(m, 1H), 2.39-2.17(m, 4H), 2.00-1.93(m, 1H), 1.87-1.76(m, 1H), 1.71-1.60 (m, 1H), 1.46 (d, J=6.4Hz, 3H), 1.12-1.03 (m, 2H), 1.02-0.97 (m, 7H), 0.89 (d, J=7.2Hz, 3H), 0.61-0.53 (m, 3H).

Compound 26a was prepared in a manner similar to that used in the preparation of intermediate F by using 1-(2,2,2-trifluoroethyl)piperazine instead of 1-Cbz-piperazine (compound D4).

Biological examples

Compound RM018 (compound A191, page 85) from WO2021091982, compound RM461 (page 115 of Figure 1) from WO2020132597, and compound RM351 (page 88 of Figure 1) from WO2020132597 are cited as reference compounds in this invention.

The applicant, following the procedure described in WO2020132597, further synthesized RM461-A as an analogue of RM461 by replacing chloroacetamide with 2-chloro-2-fluoro-acetamide, and provided comparative data to demonstrate the improvement and illustrate the technical problem solved.

Example 27

GSH reaction rate

Glutathione (GSH) is a tripeptide found in most tissues, particularly in the liver where it is present in high concentrations. It plays a crucial role in protecting cells from oxidative damage and the toxicity of exogenous electrophilic agents, as well as in maintaining redox balance. More specifically, glutathione conjugation aids in detoxification by binding electrophilic agents that could otherwise bind to proteins or nucleic acids, leading to cell damage and gene mutations.

Many potentially toxic electrophilic exogenous substances and some endogenous compounds are detoxified by conversion into corresponding glutathione S-conjugates, which consume intrinsic GSH and then diminish the detoxification effect. Some pharmaceuticals and halogenated workplace/environmental contaminants are bioactivated through this mechanism.

On the other hand, conjugation of glutathione and drug molecules in extrahepatic organs and the liver often leads to poor pharmacokinetic (PK) properties of the molecules (especially high clearance rates) and increases the likelihood of off-target reactions (potentially causing various toxicities). Therefore, strategies to minimize GSH metabolism are crucial. A short T _{_1/2} in the intrinsic GSH reaction indicates a high GSH reaction rate. Therefore, the T _{_1/2} in the intrinsic GSH reaction assay is determined for screening candidate drugs.

The reference compounds and the compounds of the present invention can potentially form conjugates with GSH via halogenation-modification or direct Michael addition. Therefore, this test was performed to examine the GSH reactivity of the listed compounds.

To determine the intrinsic GSH reactivity, 1 μM of the compound was incubated at 37 °C in 100 mM potassium phosphate buffer (pH 7.4) for 0, 0.5, 1, 2, 4, and 6 h in the presence and absence of 5 mM GSH. At the end of the specified time points, the samples were quenched with acetonitrile containing 10 mM N-ethylmaleimide and an internal standard. The quenched samples were centrifuged, and the supernatant was analyzed by LC-MS/MS for compound quantification. If the percentage of consumption after 6 hours of incubation was less than 20%, the compound was reported as stable; if the percentage of consumption was greater than 20%, the half-life was reported.

Table 1. GSH reaction rates of examples and compounds of the present invention

The above results clearly demonstrate that the reference compounds (RM461 and RM351) form conjugates with GSH, causing them to be consumed within 6 hours, while the compounds of the present invention maintain stability with less or no conjugation to GSH. In particular, comparing reference compound RM461 and its analogue RM461-A, the only difference being the substitution of chloroacetamide with 2-chloro-2-fluoroacetamide, as in the compounds of the present invention, it is surprising that such a small change can resolve the GSH toxicity problem found in the reference compounds.

Example 28

KRAS G12C-BRAF NanoBit Measurement

This assay was designed to measure the ability of the tested compound to disrupt the KRAS G12C-BRAF complex at the cellular level. We established the NanoBit cell assay in mammalian HEK293 (ATCC) cells.

HEK293 cells were grown and maintained in DMEM medium (Thermo Fisher Scientific) containing 10% fetal bovine serum and 1% penicillin/streptomycin. Both KRAS G12C and BRAF RBD were cloned into NanoBit vectors (BiBiT vector system, Promega) with SmBit-KRASG12C and BRAF RBD-LgBit orientation and co-transfected into HEK293 cells. Cells were then selected for 4 weeks with 100 μg/mL hygromycin B (10687010, Thermo Fisher) and blast fungicide (5 μg/mL) to obtain a stable cell pool.

On the day of assay, starting with a final concentration of 30 μM in DMSO, a 16-point 3-fold dilution was performed to prepare 75 nL of compound solution in 384-well assay plates. Cells were then seeded at 10,000 cells/25 μL/well in the 384-well plates. After 3 hours of incubation, 6 μL of live cell substrate (Promega) was added to each well. Luminescence was monitored at 20 minutes using the ultra384 model in Envision. Compounds that promoted the disruption of the KRAS G12C-BRAF RBD complex were identified as causing reduced luminescence compared to the DMSO control wells.

Table 2. Examples of the present invention and the activity of compounds in the KRAS G12C-BRAF NanoBit assay.

Example 29

Interaction determination of KRAS-BRAF with CYPA (50 nM)

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

Table 3. Examples of the present invention and the activity of compounds in the KRAS-BRAF-CYPA (50 nM) interaction assay.

Example 30

pERK inhibition assay

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

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

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

Table 4. Examples of the present invention and the activity of compounds in KRAS pERK inhibition assays.

Example 31

Cell viability assay

The purpose of this cell assay was to determine the effect of the tested compound on the proliferation of human cancer cell lines NCI-H358 (ATCC-CRL5807), AGS (ATCC-CRL-1739), and SW620 (ATCC-CCL-227) during a 3-day treatment period by using the Cell Counting Kit-8 to quantify the amount of NADPH present at the endpoint.

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

Table 5. Examples of the present invention and the activity of compounds in KRAS cell viability assays

Example 32

Human hepatocyte stability assay

Hepatocyte stability assays measured the rate of compound disappearance when cultured with cryopreserved human suspension hepatocytes. Each assay included positive controls of midazolam, raloxifene, and dextromethorphan. Culture consisted of 1 μM of the tested compound and a human hepatocyte suspension (1 × ^10⁶ cells/mL) in Wiliams' E medium supplemented with 10% FBS and 0.5% penicillin-streptomycin. The hepatocyte suspension was cultured at 37°C with intermittent shaking at 900 rpm in a 5% _CO₂ incubator. Consumption of the parent compound was monitored by LC-MS/MS analysis at 2, 10, 20, 40, 60, and 120 minutes after compound addition, by adding methanol containing an internal standard (2 μM tolbutamide). For human data, a CL_hep (mL/min/kg) > 16.24 was considered high clearance, and a CL_hep (mL/min/kg) < 6.96 was considered low clearance. 16.24 < CL_hep (mL/min/kg) > 6.96 indicates a moderate clearance rate.

Table 6. Human hepatocyte stability of examples and compounds of the present invention

The results clearly demonstrate that, in human hepatocyte stability assays, the reference compounds (RM461 and RM351) exhibited high clearance rates, while the compounds of the present invention maintained low to moderate clearance rates. Specifically, comparing the reference compound RM461 and its analogue RM461-A, the only difference being the substitution of chloroacetamide with 2-chloro-2-fluoroacetamide, as with the compounds of the present invention, surprisingly, such a small change can increase the stability of human hepatocytes compared to the reference compounds.

Claims (30)

1. A compound of formula (I), in ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with (dihalo _-C1-6 alkyl) carbonyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with (dihalo _-C1-6 alkyl) carbonyl; ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group. Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl, The piperidinyl group is substituted once or twice with a substituent independently selected from halogen, (dihalo _-C1-6 alkyl) carbonyl, and _C2-6 ynyl, or Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups; R ⁷ is a _C1-6 alkyl group; ^R2 is a _C1-6 alkyl group; ^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl; ^R4 is a _C1-6 alkoxy- _C1-6 alkyl group; ^R5 is H, morpholino, (halogenated _C1-6 alkyl)piperazinyl or _C1-6 alkylpiperazinyl; ^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group; ^A2 is a _C1-6 alkylene group; A ³ is O; Or its medicinal salt. 2. A compound of formula (Ia), in ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with (dihalo _-C1-6 alkyl) carbonyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with (dihalo _-C1-6 alkyl) carbonyl; ^R6 is a nitrogen-containing heterocyclic butyl group substituted with a _C2-6 ynyl group. Cycloalkyl groups substituted with formyl, _C2-6 ynyl, pyridyl _-C2-6 ynyl, or [( _C1-6 alkyl) ₂ (oxo)-λ6 ^- thionyl] _C1-6 alkyl carbonyl, The piperidinyl group is substituted once or twice with a substituent independently selected from halogen, (dihalo _-C1-6 alkyl) carbonyl, and _C2-6 ynyl, or Pyrroleyl groups substituted with ( _C1-6 alkylcarbonyl)carbonyl, (dihalo _-C1-6 alkyl)carbonyl, _C2-6 ynyl, cyano _-C1-6 alkyl, cycloalkylcarbonyl, or triazolyl _-C2-6 alkenylcarbonyl groups; R ⁷ is a _C1-6 alkyl group; ^R2 is a _C1-6 alkyl group; ^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl; ^R4 is a _C1-6 alkoxy- _C1-6 alkyl group; ^R5 is H, morpholino, (halogenated _C1-6 alkyl)piperazinyl or _C1-6 alkylpiperazinyl; ^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group; ^A2 is a _C1-6 alkylene group; A ³ is O; Or its medicinal salt. 3. The compound according to claim 1 or 2, wherein ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with a (dihalo _{- C1-6} alkyl) carbonyl group or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with a (dihalo _{-C1-6 alkyl) carbonyl group; wherein R6 is a pyrrolidinyl substituted with a (dihalo-C1-6} ^alkyl ) carbonyl group; and ^R7 is a _C1-6 alkyl group. 4. The compound according to any one of claims 1 to 3, wherein ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with chloro(fluoro)acetyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with chloro(fluoro)acetyl; wherein ^R6 is a pyrrolidinyl substituted with chloro(fluoro)acetyl; and ^R7 is methyl. 5. The compound according to any one of claims 1 to 4, wherein ^R1 is 6. The compound according to any one of claims 1 to 5, wherein ^R2 is isopropyl. 7. The compound according to any one of claims 1 to 6, wherein ^R3 is ethyl or trifluoroethyl. 8. The compound according to any one of claims 1 to 7, wherein ^R4 is methoxyethyl. 9. The compound according to any one of claims 1 to 8, wherein ^R4 is 10. The compound according to any one of claims 1 to 9, wherein R ⁵ is morpholino or methylpiperazinyl. 11. The compound according to any one of claims 1 to 9, wherein ^A1 is a compound in which bond "a" is attached to an indole ring. 12. The compound according to claim 1 or 2, wherein A2 is a dimethylmethylene group. 13. The compound according to claim 1 or 2, wherein... ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with (dihalo _-C1-6 alkyl) carbonyl or 6-oxo-2,7-diazaspiro[4.5]decane-7-yl substituted with (dihalo _-C1-6 alkyl) carbonyl; Wherein ^R6 is a pyrrolidinyl group substituted with a (dihalo _-C1-6 alkyl) carbonyl group; ^R7 is a _C1-6 alkyl group; ^R2 is a _C1-6 alkyl group; ^R3 is a _C1-6 alkyl or a halo- _C1-6 alkyl; ^R4 is a _C1-6 alkoxy- _C1-6 alkyl group; R ⁵ is morpholino or _C1-6 alkylpiperazino; ^A1 is an imidazolyl or phenylene, wherein the phenylene is substituted with a hydroxyl group; ^A2 is a _C1-6 alkylene group; A ³ is O; Or its medicinal salt. 14. The compound according to claim 13, wherein... ^R1 is ^R2 is isopropyl; ^R3 is ethyl or trifluoroethyl; ^R4 is R ⁵ is morpholino or methylpiperazino; ^A1 is where the "a" bond is attached to the indole ring; ^A2 is a dimethylmethylene group; A ³ is O; Or its medicinal salt. 15. A compound selected from: (3S)-1-[(2S)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-formyl-N-methyl-cyclobutanecarboxamide; N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-ethynyl-N-methyl-cyclobutanecarboxamide; (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-(2-oxopropionyl)pyrrolidine-3-carboxamide; (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-prop-2-ynylpyrrolidine-3-carboxamide; (3S)-1-(cyanomethyl)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; 3-[2-[dimethyl(oxo) ^-λ6 -thionyl]acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-cyclobutanecarboxamide; (3S)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-1-[(E)-3-(1,2,4-triazol-1-yl)prop-2-enoyl]pyrrolidine-3-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (3S)-1-(bicyclo[1.1.0]butane-1-carbonyl)-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-3-[2-(2-pyridyl)ethynyl]cyclobutanecarboxamide; N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-3-ethynyl-N-methyl-azacyclobutane-1-carboxamide; N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-4-ethynyl-N-methyl-piperidine-1-carboxamide; N-[(1S)-1-[[(8S,14S)-22-ethyl-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonadecane-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-4-ethynyl-4-fluoro-N-methyl-piperidine-1-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; 1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl [8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butanamide;] (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl [8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butanamide;] (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-(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[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; and (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.12,5.19,13.022,26]octadecane-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; Or its medicinal salt. 16. A compound RM461-A having the structure thereof, or a pharmaceutical salt thereof. 17. A method for preparing a compound according to any one of claims 1 to 15, the method comprising any one of the following steps: a) Using a coupling agent in the presence of a base, compound of formula (IX) is... It undergoes a coupling reaction with acid (X); b) Using a coupling agent in the presence of a base, compound (XIII) is converted into a compound. It undergoes a coupling reaction with the acid (XIV); c) Using a coupling agent in the presence of a base to convert compound (XVIII) It undergoes a coupling reaction with the acid (XIX); Wherein Q is a heterocyclic group; T is ( _C1-6 alkyl) _2- oxooxetane, _C1-6 alkyl carbonyl, dihaloC1-6 alkyl, oxooxetane _amino , _C2-6 ynyl, _C3-8 dienyl, _cyanoC1-6 alkyl, cycloalkyl, morpholinoC2-6 ynyl, oxoazacyclic butane, _pyridylC2-6 ynyl, or triazolylC2-6 alkenyl; ^R1 , ^R2 , R3, ^R4 , ^R5 , ^R6 , ^R7 , ^A1 , ^A2 , ^{and A3} are as defined in any one of claims 1 to 14; the coupling agent is _T3P , _HATU , PyBOP, or EDCI/ _HOBt ; the base is TEA, DIEPA, or DMAP. 18. The compound or pharmaceutical salt according to any one of claims 1 to 16, which is used as a therapeutically active substance. 19. A pharmaceutical composition comprising the compound according to any one of claims 1 to 16 and a therapeutically inert carrier. 20. Use of the compound according to any one of claims 1 to 16 for the treatment of KRAS G12C protein-related diseases. 21. Use of the compound according to any one of claims 1 to 16 for the treatment of diseases related to KRASG12C, G12D and G12V proteins. 22. Use of the compound according to any one of claims 1 to 16 for suppressing the interaction between RAS and downstream effectors, wherein the downstream effectors are RAF and PI3K. 23. Use of the compound according to any one of claims 1 to 16 for inhibiting the transmission of carcinogenic MAPK and PI3K signaling. 24. Use of the compound according to any one of claims 1 to 16 for the treatment or prevention of KRAS mutation-driven cancers, wherein said cancers are selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma, ovarian cancer, and endometrial cancer. 25. Use of the compound according to any one of claims 1 to 16 for the treatment or prevention of KRAS mutation-driven cancers, wherein said cancers are selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer. 26. The compound or pharmaceutical salt according to any one of claims 1 to 16, for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer. 27. Use of the compound according to any one of claims 1 to 16 for the preparation of a medicament for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer. 28. A method for treating or preventing KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer, the method comprising administering a therapeutically effective amount of a compound as defined in any one of claims 1 to 16. 29. The compound or pharmaceutical salt according to any one of claims 1 to 16, manufactured according to the method of claim 17. 30. The present invention as described above.