HK40116949A - Haloindole macrocyclic compounds for the treatment of cancer - Google Patents

Description

Halogenated indole 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 (PI3K)/protein kinase B (AKT) pathway.

KRAS proteins switch between inactive and active forms via binding to GTP and GDP, respectively. Under physiological conditions, the transition between these two states is regulated by guanine nucleotide exchange factors (GEFs), such as SOS1 (Seven Sons 1 homologue) 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 different 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

1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group,

5-oxo-2,6-diazaspiro[3.4]octyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decyl substituted with (dihalo _-C1-6 alkyl)carbonyl, or

Azahexacyclic butyloxy _C1-6 alkyl groups substituted with (dihalo- _C1-6 alkyl)carbonyl groups;

^R8 is a _C1-6 alkyl group;

R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _C1-6 alkyl) carbonyl group;

^R2 is a _C1-6 alkyl group;

^R3 is H, _C1-6 alkyl, or halogen;

^R4 is H or halogen;

R ⁵ is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R6 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R7 is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino;

^A1 is an imidazolyl group or a hydroxylated phenylene group;

^A2 is a _C1-6 alkylene group;

The condition is that ^R3 and ^R4 are not both H at the same time;

Or its medicinal salt.

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

Compounds of formula (I) or (Ia) exhibit good KRAS inhibitory activity against G12C, G12D, and G12V. In another embodiment, the compounds of the present invention demonstrate excellent cancer cell inhibition and human hepatocyte stability. 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 issue.

Attached Figure Description

Figure 1. X-ray crystallographic analysis of compound 4f1.

Figure 2. X-ray crystallographic analysis of compound 4f2.

Figure 3. X-ray crystallographic analysis of compound M2.

Detailed Implementation

definition

The term " _C1-6 alkyl" refers to a saturated, straight-chain or branched alkyl group containing one to six, particularly one to four, 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, and 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 terms “halogen” and “halogenated” are used interchangeably in this document to refer to fluorine, chlorine, bromine, or iodine.

The term "halogenated indole" refers to an indole in which one or more hydrogen atoms have been replaced by the same or different halogen atoms.

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, especially 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 "piperidinyl" refers to a divalent piperidinyl group.

The term "pyridinealkylene" refers to a divalent pyridine alkyl group.

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

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

The term "azacyclobutyryloxy" means aazacyclobutyryl-O-.

The term "dimethylmethylene" means

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 hydroxy protecting groups.

Those skilled in the art will understand that the following structures of compounds of formula (Ia) and (Ia’) are identical (especially for the chiral center):

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

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

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

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

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

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

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

KRAS inhibitors

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

in

^R1 is

1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group,

5-oxo-2,6-diazaspiro[3.4]octyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decyl substituted with (dihalo _-C1-6 alkyl)carbonyl, or

Azahexacyclic butyloxy _C1-6 alkyl groups substituted with (dihalo- _C1-6 alkyl)carbonyl groups;

^R8 is a _C1-6 alkyl group;

R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _C1-6 alkyl) carbonyl group;

^R2 is a _C1-6 alkyl group;

^R3 is H, _C1-6 alkyl, or halogen;

^R4 is H or halogen;

R ⁵ is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R6 is a _C1-6 alkoxy- _C1-6 alkyl group;

R ⁷ is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino;

^A1 is an imidazolyl group or a hydroxylated phenylene group;

^A2 is a _C1-6 alkylene group;

The condition is that ^R3 and ^R4 are not both H at the same time;

Or its medicinal salt.

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

in

^R1 is

1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group,

5-oxo-2,6-diazaspiro[3.4]octyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decyl substituted with (dihalo _-C1-6 alkyl)carbonyl, or

Azahexacyclic butyloxy _C1-6 alkyl groups substituted with (dihalo- _C1-6 alkyl)carbonyl groups;

^R8 is a _C1-6 alkyl group;

R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _C1-6 alkyl) carbonyl group;

^R2 is a _C1-6 alkyl group;

^R3 is H, _C1-6 alkyl, or halogen;

^R4 is H or halogen;

R ⁵ is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R6 is a _C1-6 alkoxy- _C1-6 alkyl group;

^R7 is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino;

^A1 is an imidazolyl group or a hydroxylated phenylene group;

^A2 is a _C1-6 alkylene group;

The condition is that ^R3 and ^R4 are not both H at the same time;

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]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, or

5-oxo-2,6-diazaspiro[3,4]octyl groups substituted with (dihalo _-C1-6 alkyl)carbonyl groups;

^R8 is a _C1-6 alkyl group;

R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _-C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _-C1-6 alkyl) carbonyl 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

1-oxo-2,7-diazaspiro[4.4]nonane-2-yl substituted with chloro(fluoro)acetyl,

1-oxo-2,8-diazaspiro[4.5]decane-2-yl substituted with chloro(fluoro)acetyl,

1-oxo-2,9-diazaspiro[5.5]undecane-2-yl substituted with chloro(fluoro)acetyl, or

5-oxo-2,6-diazaspiro[3,4]octane-6-yl substituted with chloro(fluoro)acetyl;

^R8 is a methyl group;

R ⁹ is a piperidinyl group that has been substituted twice with fluorine and chloro(fluoro)acetyl groups, or a pyrroleyl group that has been substituted with chloro(fluoro)acetyl groups.

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 (5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, (5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, 8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl, 9 -[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl, 2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl, 1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-4-piperidinyl or 1-[(2R)-2-chloro-2-fluoro-acetyl]pyrrolidine-3-yl.

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 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 H, methyl or fluorine.

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 ^R3 is fluorine.

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

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 ^R4 is H.

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 ethyl or 2,2,2-trifluoroethyl.

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 1-methoxyethyl.

A further embodiment of the present invention is (xiii) a compound of formula (I) or (Ia) according to any one of (i) to (xii), or a pharmaceutical salt thereof, wherein ^R7 is morpholino, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or 4-methylpiperazin-1-yl.

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

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

^R1 is

1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, or

5-oxo-2,6-diazaspiro[3,4]octyl groups substituted with (dihalo _-C1-6 alkyl)carbonyl groups;

^R8 is a _C1-6 alkyl group;

R ⁹ is a piperidinyl group that has undergone two substitutions with a halogen and a (dihalogenated _C1-6 alkyl) carbonyl group, or a...

(Dihalogenated _C1-6 alkyl)carbonyl-substituted pyrrolidinyl;

^R2 is a _C1-6 alkyl group;

^R3 is a halogen;

^R4 is H;

R ⁵ is a _C1-6 alkyl or a halo- _C1-6 alkyl;

^R6 is a _C1-6 alkoxy- _C1-6 alkyl group;

R ⁷ is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino;

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

^A2 is a _C1-6 alkylene group;

Or its medicinal salt.

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

^R1 is (5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, (5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, 8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl, 9 -[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl, 2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl, 1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-4-piperidinyl or 1-[(2R)-2-chloro-2-fluoro-acetyl]pyrrolidine-3-yl;

^R2 is isopropyl;

^R3 is fluorine;

^R4 is H;

R ⁵ is ethyl or 2,2,2-trifluoroethyl;

R ⁶ is (1S)-1-methoxyethyl;

R ⁷ is morpholino, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or 4-methylpiperazin-1-yl;

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

^A2 is a dimethylmethylene group;

Or its medicinal salt.

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

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [28-octadec-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [28-octadec-1(25),2,5(28),19,22(26),23-hexene-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]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17,24-trimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [28-octadec-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^{23 ,27]} [26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

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]-17,17,24-trimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide;

1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.5]nonane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

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

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

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5S)-2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5R)-2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5R)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5S)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-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]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [28-octadec-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-26-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^{23 ,27]} [26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26 ] [28-octadec-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide;

1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidine-4-carboxamide;

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

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

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

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-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)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidine-4-carboxamide;

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidine-4-carboxamide;

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-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)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide; and

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide;

Or its medicinal salt.

Another embodiment of the present invention relates to (xix) a compound having the following structure:

Another embodiment of the present invention relates to (xx) a method for preparing a compound according to any one of (i) to (xix), the method comprising any one of the following steps:

a) In the presence of a coupling agent and a base, compound (II) is formed.

It undergoes a coupling reaction with acid (III) to form a compound of formula (I);

b) In the presence of a coupling agent and a base, compound of formula (VI) is...

It undergoes a coupling reaction with acid (VII) to form compound (VIII).

c) To make compound (XI) in the presence of a coupling agent and a base.

It undergoes a coupling reaction with acid (XII) to form (XIII).

Compounds,

Wherein Q is an unsubstituted or substituted piperidinyl or pyrrolidine; T is a dihalogenated _C1-6 alkyl; ^R1 , R2, ^R3 , ^R4 , ^R5 , ^R6 , ^R7 , ^{A1 and A2} are as defined in any of (i) to (xvii); the coupling agent in steps a) to c) is _T3P , HATU, PyBOP or EDCI/HOBt; the base in steps a) to c) is TEA, DIEPA or DMAP.

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

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

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

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

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

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

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

Another embodiment of the invention is (xxviii) the use of the compound according to any one of (i) to (xix) 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 (xxix) a compound or pharmaceutical salt according to any one of (i) to (xix) 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 (xxx) the use of the compound according to any one of (i) to (xix) 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 (xxxi) 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 (xix).

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

Pharmaceutical composition and administration

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

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

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

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

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

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

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

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

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

The following 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, in particular ^R1 to ^R7 , ^A1 and ^A2 , are as defined above. Furthermore, unless explicitly stated otherwise, all reactions, reaction conditions, abbreviations and symbols have meanings well known to those skilled in the art of organic chemistry.

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

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

PG is a protecting group, such as Boc and Cbz.

Alternatively, compound (V) can be obtained by coupling acid (IV) with compound (II) 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 (V) 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 (VI). Compound (VIII) can be obtained by coupling acid (VII) with compound (VI) in the presence of a base such as TEA, DIEPA, or DMAP using a coupling agent such as _T3P , HATU, PyBOP, or EDCI/HOBt.

Where PG is a protecting group, such as Boc and Cbz; T is a dihalogenated _C1-6 alkyl group; and Q is an unsubstituted or substituted piperidinyl or pyrrolidine group.

Alternatively, compound (X) can be obtained by coupling an acid (IX), compound (VI), and a coupling agent such as _T3P , HATU, PyBOP, or EDCI/HOBt in the presence of a base such as TEA, DIEPA, or DMAP. Deprotection of compound (X) 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 (XI). Compound (XIII) can be obtained by coupling an acid (XII) with compound (XI) 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) In the presence of a coupling agent and a base, compound (II) is formed.

They undergo coupling reactions to form compounds of formula (I);

b) In the presence of a coupling agent and a base, compound of formula (VI) is...

It undergoes a coupling reaction with acid (VII) to form compound (VIII).

c) To make compound (XI) in the presence of a coupling agent and a base.

It undergoes a coupling reaction with acid (XII) to form (XIII).

Compounds,

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; T is a dihalogenated _C1-6 alkyl; and Q is an unsubstituted or substituted piperidinyl or pyrrolidine alkyl.

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

Example

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

abbreviation

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

The abbreviations used in this article are as follows:

ACN Acetonitrile

aq. water-based

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

(Boc) _₂O ditert-butyl dicarbonate

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

CDCl ₃ : Deuterated chloroform

_CD3 OD: Deuterated methanol

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

DIEPA: N,N-Diethylpropylamine

DIBAL-H diisobutylaluminum hydride

DMAP: 4-Dimethylaminopyridine

DMF: Dimethylformamide

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-triazolidine]

[4,5-b]pyridinium 3-oxide hexafluorophosphate)

hr: hours

HPLC: High Performance Liquid Chromatography

HOBt: N-hydroxybenzotriazole

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

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

LDA (Lithium diisopropylamino)

MS (ESI): Mass spectrometry (electrospray ionization)

min(s) minutes

NMM N-methylmorpholine

NMR: Nuclear Magnetic Resonance

NMO 4-methylmorpholine N-oxide

obsd. Observation value

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

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

preparative high performance liquid chromatography (prep-HPLC)

PyBOP: Benzotriazole-1-yloxytripyrrolidinephosphine hexafluorophosphate

RT or rt: Room temperature

sat. saturated

SFC Supercritical Fluid Chromatography

TEA: Triethylamine

TFA: Trifluoroacetic acid

THF: Tetrahydrofuran

TEA: 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 using chiral columns (Daicel chiralpak IC, 5 μm, 30 × 250 mm), AS (10 μm, 30 × 250 mm), or AD (10 μm, 30 × 250 mm), employing the Mettler Toledo Multigram III SFC system, Waters 80Q preparative SFC, or Thar 80 preparative SFC. Solvent systems were _CO2 and IPA (0.5% TEA in IPA solution) or _CO2 and MeOH (0.1% _NH3 · _H2O in MeOH solution), with a back pressure of 100 bar 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 A1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-3-pyridyl]-4-methyl-piperazine

Title intermediate A was prepared according to the following scheme:

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

Add 4,4'-di-tert-butyl-2,2'-bipyridine (372.7 mg, 1.39 mmol) and [Ir(OMe)(COD)] _₂ (306.3 mg, 0.460 mmol) to a solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (compound A1, 2.0 g, 9.26 mmol) and bis(pinacol)diboron (3.5 g, 13.9 mmol) in THF (30 mL). Stir the mixture at 75 °C for 16 hours under _N₂ . Filter the mixture and concentrate the filtrate under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-20%) to give 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)pyridine (compound A2, 2.4 g), which was a yellow oil. ^¹H NMR (400 MHz, CDCl₃) δppm 8.91 (d, J = 1.4 Hz, 1H), 8.21 (d, J = 1.4 Hz, 1H), 4.95 (q, J = 6.5 Hz, 1H), 3.30 (s, 3H), 1.49 (d, J = 6.5 Hz, 3H), 1.35 (s, 12H).

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

N-iodosuccinimide (4.1 g, 18.27 mmol) was added to a solution of 3-bromo-2-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)pyridine (compound A2, 2.5 g, 7.3 mmol) in ACN (40 mL). The mixture was stirred at 90 °C for 40 h under _N2 protection. The reaction was quenched with saturated _Na2SO3 solution ( ₄₀ mL), and the reaction mixture was extracted 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 A3, 660 mg), which was 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 (intermediate A5)

Cesium carbonate (1.6 g, 4.83 mmol), (R)-BINAP (60.1 mg, 0.1 mmol), and palladium(II) acetate (43.3 mg, 0.19 mmol) were added to a solution of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (compound A3, 660 mg, 1.9 mmol) and 1-Cbz-piperazine (compound A4, 425.1 mg, 1.9 mmol) in toluene (10 mL). The mixture was stirred at 100 °C for 12 hours under _N2 . The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (EA/PE: 0-50%) to give 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (intermediate A5, 740 mg), which is a yellow solid. MS calculated value 434.1 (MH ⁺ ); measured value 434.1 (MH ⁺ ).

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

KOAc (418.0 mg, 4.26 mmol) and Pd(dppf)Cl₂ (124.7 mg, 0.170 mmol) were added to a solution of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (intermediate A5, 740 mg, 1.7 mmol) and bis(pinacol) _diboron (519.2 mg, 2.04 mmol) in toluene (12 mL). The reaction mixture was stirred at 90 °C for 12 hours under _N₂ protection. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography to give 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-3-pyridyl]-4-methyl-piperazine (intermediate A, 470 mg), a brown solid. MS calculated value 482.3 (MH ⁺ ); measured value 482.2 (MH ⁺ ).

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 hours. 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), 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), which was 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 methyl (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 silica gel 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-dioxaneborane-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), 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 chromatography 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), which is a yellow oil. MS: calculated value 690 (MH ⁺ ); measured value 690.4 (MH ⁺ ).

Intermediate C

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

Intermediate C was prepared according to the following scheme:

Step 1: Preparation of (4-bromothiazol-2-yl)methanol (compound C2)

Sodium borohydride (1.7 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 was quenched with water (300 mL) at 0 °C, and the reaction mixture was 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 C2, 6 g), which was a colorless oil.

Step 2: Preparation of 4-bromo-2-(bromomethyl)thiazole (compound C3)

At 0 °C, CBr ₄ (15.4 g, 46.38 mmol) and triphenylphosphine (12.1 g, 46.38 mmol) were added to a solution of (4-bromothiazol-2-yl)methanol (compound C2, 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 C3, 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 C5)

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 C4, 4.3 g, 23.45 mmol) in THF (60 mL). After addition, the mixture was stirred at -78 °C for 0.5 h. At -78 °C, 4-bromo-2-(bromomethyl)thiazole (compound C3, 5.4 g, 21.02 mmol) was added to the above mixture, and the mixture was stirred for another 1 h. The reaction was quenched with saturated _NH4Cl solution (100 mL), and the reaction mixture was extracted 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 under vacuum. The residue was purified by reversed-phase chromatography to give 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (compound C5, 3.6 g), which was a yellow oil. MS calculated value 360 (MH ⁺ ); measured value 359.9 (MH ⁺ ).

Step 4: Preparation of methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound C6)

Hydrochloric acid (66.6 mL, 0.3 M) was added to a solution of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl]thiazole (compound C5, 3.6 g, 10 mmol) in ACN (20 mL). The mixture was stirred at 25 °C for 2 h. The mixture was alkalized with saturated _NaHCO3 solution until pH 8. The mixture was extracted twice with EtOAc (80 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give methyl (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound C6, 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 C7)

Triethylamine (2.9 g, 29.23 mmol) and (Boc) _₂O (3.8 g, 17.54 mmol) were added to a solution of (2S)-2-amino-3-(4-bromothiazol-2-yl)propionate (compound C6, 3.1 g, 11.69 mmol) in DCM (40 mL). After stirring at 30 °C for 12 hours, the mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in petroleum ether (0–30%) to give methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound C7, 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 C8)

Lithium hydroxide (0.4 mL, 43.81 mmol) was added to a solution of methyl (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound C7, 3.2 g, 8.76 mmol) in THF (30 mL), methanol (2 mL), and water (10 mL). After stirring at 25 °C for 1 hour, the reaction mixture was acidified with 1 M HCl solution until pH 5. The mixture was extracted twice with EtOAc (40 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionate (compound C8, 3.1 g) as a yellow oil. MS calculated value 373 (MNa ⁺ ); measured value 372.9 (MNa ⁺ ).

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

At 0 °C, a solution of (2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionic acid (compound C8, 3.1 g, 8.83 mmol) in DCM (50 mL) was added with methyl (3S)-hexahydropyridazine-3-carboxylate hydrochloride (compound C9, 2.4 g, 13.24 mmol), EDCI (3.4 g, 17.65 mmol), 1-hydroxybenzotriazole (238.5 mg, 1.77 mmol), and NMM (9.92 mL, 88.26 mmol). After stirring at 25 °C for 1 hour, the reaction mixture was diluted with water (60 mL) and extracted 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 C, 2.4 g). MS calculated value 477 (MH ⁺ ), measured value 476.9 (MH ⁺ ).

Intermediate D

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

Title intermediate D is prepared according to the following scheme:

Step 1: Preparation of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylprop-1-one (compound D3)

At 0 °C, _a SnCl₄ solution (97.2 mL, 121.5 mmol) was slowly added to a mixture of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropionyl chloride (compound D1, 35.0 g, 116.8 mmol) in DCM (400 mL). After stirring at -40 °C for 0.5 hours, a solution of 5-bromo-6-fluoro-1H-indole (compound D2, 25.0 g, 116.8 mmol) in DCM (200 mL) was added dropwise, and the mixture was stirred at -40 °C for 15 min. After the reaction was complete, it was quenched with a saturated aqueous solution of _NaHCO₃ (800 mL), and the reaction mixture was extracted twice with EtOAc (900 mL). The combined organic layers were washed with brine (700 _mL ), dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was ground together with the solution (100 mL, petroleum ether: ethyl acetate = 8:1) and filtered. The filter cake was dried under vacuum to give 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylprop-1-one (compound D3, 50.0 g), a yellow solid. MS calculated value 552.1 (MH ⁺ ); measured value 552.1 (MH ⁺ ).

Step 2: Preparation of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound D4)

At 0 °C, LiBH₄ (48.4 mL, 193.49 mmol, 4 M, in THF) was added dropwise to a mixture of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylprop-1-one (compound _D3 , 50.0 g, 90.49 mmol) in THF (600 mL). The mixture was stirred at 70 °C for 24 h under a nitrogen atmosphere. After the reaction was complete, it was quenched by the slow addition of water (600 mL) at 0 °C, and the reaction mixture was extracted twice with EtOAc (600 mL). The combined organic layers were washed with brine (600 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (EtOAc = 20%–33% in PE) to give [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound D4, 46.0 g), a white solid. MS calculated value 538.1 (MH ⁺ ); measured value 538.2 (MH ⁺ ).

Step 3: Preparation of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound D5)

Silver trifluoromethanesulfonate (20.3 g, 78.88 mmol) was added to a mixture of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound D4, 35.4 g, 65.73 mmol) and _iodine (18.4 g, 72.3 mmol) in THF (400 mL) at 0 °C. The mixture was stirred at 0 °C for 10 min. After the reaction was complete, it was quenched with a saturated aqueous solution of _Na₂SO₃ (400 mL) and EtOAc (400 mL), and the reaction mixture was filtered. The organic layer was washed with brine (100 _mL ), dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (EtOAc = 0%–2.5% in PE) to give [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound D5, 43.0 g), a yellow solid. MS calculated value 664.0 (MH ⁺ ); measured value 664.1 (MH ⁺ ).

Step 4: Preparation of 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D6)

Potassium phosphate (15.7 g, 73.92 mmol) and Pd(dppf)Cl₂ (920 mg, 1.26 mmol) were added to a mixture of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound D5, 16.7 g, 25.13 mmol) and 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-3- _pyridyl ]piperazine-1-carboxylic acid benzyl ester (intermediate A, 16.7 g, 34.69 mmol) in a mixed solution of 1,4-dioxane (270 mL)/toluene (90 mL)/water (90 mL). The mixture was stirred at 70 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the mixture was filtered and concentrated under vacuum. The residue was purified by silica gel column chromatography (EtOAc = 20%–50% in PE) to give 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound D6, 19.5 g), a white solid. MS calculated value 891.3 (MH ⁺ ); measured value 891.3 (MH ⁺ ).

Step 5: Preparation of 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D7)

To a solution of 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound D6, 14.5 g, 16.26 mmol) and _Cs₂CO₃ (15.9 _g , 48.77 mmol) in DMF (200 mL), 2,2,2-trifluoroethyl trifluoromethanesulfonate (37.7 g, 162.56 mmol) was added dropwise at 0 °C, and the mixture was stirred at 20 °C for 12 h. After the reaction was complete, EtOAc (70 mL) and water (100 mL) were added, and the layers were separated. The aqueous phase was extracted twice with EtOAc (70 mL). The combined organic layers were washed four times with brine (100 mL), dried over _{Na₂SO₄ ,} filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography to give 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D7, 8.0 g, PEAK 1, fast elution), which was a yellow oil. MS calculated value 973.3 (MH ⁺ ); measured value 973.2 (MH ⁺ ).

Step 6: Preparation of 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D8)

Cesium fluoride (8.2 g, 53.9 mmol) was added to a solution of 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D7, 10.5 g, 10.78 mmol) in DMF (130 mL), and the mixture was stirred at 60 °C for 24 h. After the reaction was complete, EtOAc (100 mL) and water (100 mL) were added, and the layers were separated. The aqueous phase was extracted twice with EtOAc (100 mL). The combined organic layers were washed three times with brine (80 _mL ), dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give the residue. The residue was purified by silica gel column chromatography (EtOAc = 25%–66% in PE) to give 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D8, 6.5 g), as a yellow solid. MS calculated value 735.2 (MH ⁺ ); measured value 735.1 (MH ⁺ ).

Step 7: Preparation of 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D9)

Pd(dppf)Cl₂ (537.1 mg, 0.73 mmol) was added to a solution of 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound _D8 , 5.4 g), bis(pinacol)diboron (2.8 g, 11.01 mmol), and potassium acetate (1.2 mL, 18.35 mmol) in toluene (70 mL). The mixture was degassed and purged three times under a nitrogen atmosphere, and stirred at 90 °C for 12 h. After the reaction was complete, the mixture was cooled to room temperature. The reaction mixture was filtered, and the filtrate was concentrated under vacuum to give the residue. The residue was purified by silica gel column chromatography (EtOAc = 25%–66% in PE) to give 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D9, 5.2 g), which is a yellow oil. MS calculated value 783.3 (MH ⁺ ); measured value 783.3 (MH ⁺ ).

Step 8: Preparation of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound D10)

Under a nitrogen atmosphere, K₃PO₄ was added to a mixture of (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate C, 2.7 g, 5.69 mmol) and 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound _D9 , 4.9 g, 6.32 mmol) in toluene (60 mL)/1,4-dioxane (20 mL)/water (20 mL ₎ . (3.4 g, 15.81 mmol) and Pd(dtbpf) _Cl₂ (412.2 mg, 0.63 mmol). The mixture was stirred at 70 °C for 12 hours. After the reaction was complete, the mixture was concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography (EtOAc = 10%–75% in PE) to give methyl (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylate (compound D10, 3.6 g), a brown solid. MS calculated value 1053.4 (MH ⁺ ); measured value 1053.3 (MH ⁺ ).

Step 9: Preparation of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid (compound D11)

Trimethyltin alcohol (2.4 g, 13.67 mmol) was added to a solution of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]-2-(tert-butoxycarbonylamino)-propionyl]-hexahydropyridazine-3-carboxylate (compound D10, 3.6 g, 3.42 mmol) in DCE (50 mL), and the mixture was stirred at 60 °C for 12 h. After the reaction was complete, EtOAc (80 mL) and water (60 mL) were added, and the layers were separated. The aqueous phase was extracted twice with EtOAc (80 mL). The combined organic layers were washed with brine (100 mL ₎ , dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazin-3-carboxylic acid (compound D11, 4.3 g), a brown solid. MS calculated value 1039.4 (MH ⁺ ); measured value 1039.2 (MH ⁺ ).

Step 10: Preparation of 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D12)

At 0 °C, DIEA (14.4 mL, 82.76 mmol), EDCI (11.9 g, 62.07 mmol), and 1-hydroxybenzotriazole (1.4 g, 10.35 mmol) were added to a mixture of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid (compound D11, 4.3 g, 4.14 mmol) in DCM (430 mL). The mixture was stirred at 15 °C for 12 hours. After the reaction was complete, the mixture was concentrated under vacuum, then diluted with water (80 mL), and extracted twice with EtOAc (80 mL). The combined organic layers were washed with brine (80 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (EtOAc = 25%–66% in PE) to give 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D12, 3.1 g), which is a yellow gel. MS calculated value 1021.4 (MH ⁺ ); measured value 1021.2 (MH ⁺ ).

Step 11: Preparation of N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl] tert-butyl carbamate (compound D13)

To 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26]} A mixture of 1,8-octadecano-1(25),2,5(28),19,22(26),23-hexaden-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D12, 3.1 g, 3.04 mmol) and an aqueous solution of formaldehyde (775.0 mg, 9.55 mmol) in methanol (150 mL) was supplemented with activated carbon-supported Pd(OH) _₂ (2.79 g, 3.97 mmol). The mixture was degassed and purged three times with _H₂ . The mixture was hydrogenated at 30 °C for 18 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated under vacuum to obtain N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound D13, 2.6 g), which is a brown solid. MS calculated value 901.3 (MH ⁺ ); measured value 901.3 (MH ⁺ ).

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

TFA (14.0 mL, 181.72 mmol) was added to a mixture of N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound D13, 2.6 g, 2.89 mmol) in DCM (18 mL). The mixture was stirred at 15°C for 0.5 h. After the reaction was complete, the mixture was concentrated under vacuum and diluted with saturated _NaHCO3 (30 mL), and extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine (50 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D, 2.0 g), a yellow solid, which was used directly in the next step. MS: Calculated value 801.3 (MH ⁺ ); Measured value 801.2 (MH ⁺ )

Intermediate E

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

Similar to the preparation of intermediate D, the title compound was prepared by using 5-bromo-4-fluoro-1H-indole instead of 5-bromo-6-fluoro-1H-indole (compound D2).

intermediate F

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

Similar to the preparation of intermediate D, the title compound was prepared by using 5-bromo-6-methyl-1H-indole instead of 5-bromo-6-fluoro-1H-indole (compound D2).

intermediate G

(8S,14S)-8-amino-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-9,15-dione

Compound intermediate G was prepared according to the following scheme:

Step 1: Preparation of (3S)-1-[(2S)-3-[3-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound G1)

To 4-[(5M)-5-[5-bromo-1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D8, 950.0 mg, 1.29 mmol) and (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[3-(4 Methyl 4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-5-triisopropylsilyloxy-phenyl]propionyl]hexahydropyridazine-3-carboxylate (intermediate B, 1.07 g, 1.55 mmol) was added to a mixture of 1,4-dioxane (25 mL) and water (5 mL) with sodium carbonate (342.2 mg, 3.23 mmol) and Pd(dtbpf) _Cl₂ (84.2 mg, 0.13 mmol). The mixture was stirred at 85 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by reverse-phase rapid column chromatography and concentrated under vacuum to give methyl (3S)-1-[(2S)-3-[3-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (compound G1, 1.3 g), a brown solid. MS calculated value 1218.6 (MH ⁺ ); measured value 1218.5 (MH ⁺ ).

Step 2: Preparation of (3S)-1-[(2S)-3-[3-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid (compound G2)

Me 3 SnOH (652.9 mg, 3.61 mmol) was added to a mixture of (3S)-1-[(2S)-3-[3-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine- ₃ -carboxylate (compound G1, 1.1 g, 0.9 mmol) in a DCE (20 mL). The mixture was stirred at 60 °C for 12 hours. After the reaction was complete, the mixture was diluted with water (100 mL) and extracted twice with EtOAc (100 mL). The combined organic layers were washed with brine (150 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give (3S)-1-[(2S)-3-[3-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylic acid (compound G2, 1.5 g), a yellow solid, which was used directly in the next step. MS calculated value 1204.6 (MH ⁺ ); measured value 1204.5 (MH ⁺ ).

Step 3: Preparation of 4-[5-[(8S,14S)-8-(tert-butoxycarbonylamino)-25-fluoro-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecano-1(26),2,4,6(29),20,23(27),24-heptaen-(21M)-21-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound G3)

At 0 °C, DIEA (4.4 mL, 24.95 mmol), EDCI (3.6 g, 18.71 mmol), and HOBt (421.4 mg, 3.12 mmol) were added to a mixture of (3S)-1-[(2S)-3-[3-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-pyridinyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5-yl]-5-triisopropylsilyloxy-phenyl]-2-(tert-butoxycarbonylamino)-propionyl]hexahydropyridazine-3-carboxylic acid (compound G2, 1.5 g, 1.25 mmol) in DCM (150 mL). After stirring at 15°C for 12 hours, the reaction mixture was concentrated under vacuum and diluted with water (50 mL), and extracted three _times with EtOAc (50 mL). The combined organic layers were washed with brine (100 mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by column chromatography (EtOAc = 25%–50% in PE) to give 4-[5-[(8S,14S)-8-(tert-butoxycarbonylamino)-25-fluoro-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27] [26),2,4,6,20,23,27,24-heptaen-(21M)-21-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound G3, 1.1 g), is a yellow solid. MS calculated value 1186.4 (MH ⁺ ); measured value 1186.5 (MH ⁺ ).

Step 4: Preparation of N-[(8S,14S)-25-fluoro-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound G4)

To 4-[5-[(8S,14S)-8-(tert-butoxycarbonylamino)-25-fluoro-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27] A mixture of 29-carbon-1(26),2,4,6(29),20,23(27),24-heptaen-(21M)-21-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound G3, 890.0 mg, 0.75 mmol) and formaldehyde (182.6 mg, 2.25 mmol) in methanol (15 mL) was supplemented with activated carbon-supported Pd(OH) _₂ (700.0 mg, 1.0 mmol). The mixture was degassed and purged three times with _H₂ . The mixture was hydrogenated at 30 °C for 18 hours. The mixture was filtered, and the filtrate was concentrated under vacuum to obtain N-[(8S,14S)-25-fluoro-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecano-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound G4, 780.0 mg), a colorless gel that was used directly in the next step. MS calculated value 1066.5 (MH ⁺ ); measured value 1066.7 (MH ⁺ ).

Step 5: Preparation of N-[(8S,14S)-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl] tert-butyl carbamate (compound G5)

At 0 °C, N-[(8S,14S)-25-fluoro-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-4-triisopropylsilyloxy-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 A mixture of tert-butyl carbamate (compound G4, 780.0 mg, 0.73 mmol) in THF (10 mL) was mixed with tetrabutylammonium fluoride (0.8 mL, 0.8 mmol, 1 M, in THF). After stirring at 0 °C for 0.5 h, the reaction mixture was diluted with water (20 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed twice with brine (30 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain N-[(8S,14S)-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^{10 ,14} .0 ^23,27 [26),2,4,6,20,23,27,24-heptaen-8-yl]tert-butyl carbamate (compound G5, 700.0 mg, 0.77 mmol), a yellow solid, was used directly in the next step. MS calculated value 910.4 (MH ⁺ ); measured value 910.7 (MH ⁺ ).

Step 6: Preparation of (8S,14S)-8-amino-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecan-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate G)

TFA (6.0 mL, 77.88 mmol) was added to a mixture of N-[(8S,14S)-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecano-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (compound G5, 700.0 mg, 0.77 mmol) in DCM (10 mL). After stirring at 15°C for 0.5 h, the reaction mixture was concentrated in vacuum and diluted with saturated NaHCO3 ( ₁₅ mL), and extracted twice with EtOAc (50 mL). The combined organic layers were washed with brine (20 mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give (8S,14S)-8-amino-25-fluoro-4-hydroxy-(21M) _-21- [2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ] 29-carbon-1(26),2,4,6(29),20,23(27),24-heptaen-9,15-dione (intermediate G, 600.0 mg), a yellow solid, which was used directly in the next step. MS calculated value 810.3 (MH ⁺ ); measured value 810.4 (MH ⁺ ).

intermediate H

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

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

Intermediate I

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

The compound was prepared according to the following scheme:

Step 1: Preparation of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (compound I2)

Cesium carbonate (8.1 g, 24.87 mmol), (R)-binap (0.3 g, 0.5 mmol), and palladium(II) acetate (0.1 g, 0.5 mmol) were added to a mixture of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (compound I1, 3.4 g, 9.94 mmol) and morpholine (0.9 g, 9.94 mmol) in toluene (50 mL). The mixture was stirred at 100 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the mixture was filtered and concentrated under vacuum. The residue was purified by column chromatography (EtOAc = 10%–25% in PE) to give 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (compound I2, 2100.0 mg), which was a colorless oil. MS calculated value 301.1 (MH ⁺ ); measured value 301.1 (MH ⁺ ).

Step 2: Preparation of 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-3-pyridyl]morpholine (compound I3)

KOAc (1.7 g, 17.43 mmol) and Pd(dppf)Cl₂ (0.2 g, 0.35 mmol) were added to a mixture of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]morpholine (compound I₂, 2.1 g, 6.97 mmol) and bis(pinacol) _diboron (2.1 g, 8.37 mmol) in 1,4-dioxane (25 mL). The mixture was stirred at 90 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the reaction mixture was filtered and the filtrate was concentrated under vacuum. The residue was subjected to column chromatography (EtOAc = 10%–25% in PE) to give 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-3-pyridyl]morpholine (compound I3, 1100.0 mg), a brown solid. MS: calculated value 349.2 (MH ⁺ ); measured value 349.2 (MH ⁺ ).

Step 3: Preparation of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound I4)

Potassium phosphate (1.8 g, 8.64 mmol) and Pd(dppf)Cl₂ (0.22 g, 0.29 mmol) were added to a mixture of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound _D6 , 1.9 g, 2.94 mmol) in 1,4-dioxane (25 mL) and water (5 mL). After the reaction was complete, the reaction mixture was filtered, and the filtrate was diluted with water (50 mL) and extracted three times with EtOAc (100 mL). The combined organic phases were washed with brine (200 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by column chromatography to give [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound I4, 1.4 g), which was a yellow gel. MS calculated value 758.2 (MH ⁺ ); measured value 758.2 (MH ⁺ ).

Step 4: Preparation of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound I5)

At 0 °C, 2,2,2-trifluoroethyl trifluoromethanesulfonate (4.4 g, 18.9 mmol) was added dropwise to a solution of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound I4, 1.4 g, 1.89 mmol) and cesium carbonate (1.8 g, 5.67 mmol) in DMF (30 mL), and the mixture was stirred at 30 °C for 16 hours. After the reaction was complete, EtOAc (200 mL) and water (200 mL) were added, and the layers were separated. The aqueous phase was extracted twice with EtOAc (100 mL). The combined organic layers were washed three times with brine (100 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by column chromatography (EtOAc = 25%–66% in PE) to give [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound I5, 800.0 mg, PEAK 1, fast elution), a brown solid. MS calculated value 840.1 (MH ⁺ ); measured value 840.1 (MH ⁺ ).

Step 5: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound I6)

Cesium fluoride (0.7 g, 4.93 mmol) was added to a solution of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound I5, 0.8 g, 0.95 mmol) in DMF (10 mL), and the mixture was stirred at 60 °C for 15 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. EtOAc (70 mL) and water (100 mL) were added, and the layers were separated. The aqueous phase was extracted twice with EtOAc (70 mL). The combined organic layers were washed four times with brine (80 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give the residue. The residue was purified by column chromatography (EtOAc = 10%–66% in PE) to give 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound I6, 430.0 mg), as a yellow gel. MS: calculated value 602.1 (MH ⁺ ); measured value 602.2 (MH ⁺ ).

Step 6: Preparation of 3-[6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound I7)

Potassium acetate (0.1 mL, 1.78 mmol) and Pd(dppf)Cl₂ (52.2 mg, 0.07 mmol) were added to a solution of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-prop- ₁ -ol (compound I6, 430.0 mg, 0.71 mmol) and bis(pinacol)diboron (199.3 mg, 0.79 mmol) in toluene (5 mL). The mixture was degassed by purging nitrogen for 2 min, and then stirred at 90 °C for 15 h under a nitrogen atmosphere. After the reaction was complete, the reaction mixture was filtered and the filtrate was concentrated under vacuum to give the residue. The residue was subjected to column chromatography (EtOAc = 10%–66% in PE) to give 3-[6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound I7, 390.0 mg), as a colorless gel. MS: calculated value 650.3 (MH ⁺ ); measured value 650.3 (MH ⁺ ).

Step 7: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound I8)

Add K₃PO₄ to a mixture of 3-[6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound I7, 390.0 mg, 0.6 mmol) and (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate C, 291.6 mg, 0.61 mmol) in toluene ( ₄ mL), 1,4-dioxane (1.3 mL), and water (1.3 mL) _. (320.06 mg, 1.51 mmol) and Pd(dtbpf) _Cl₂ (39.1 mg, 0.06 mmol). The mixture was stirred at 70 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the reaction mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by column chromatography (EtOAc = 10%–75% in PE) to give (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylate (compound I8, 200.0 mg), a brown solid. MS calculated value 920.3 (MH ⁺ ); measured value 920.3 (MH ⁺ ).

Step 8: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound I9)

Trimethyltin alcohol (133.6 mg, 0.74 mmol) was added to a solution of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylate (compound I8, 170.0 mg, 0.18 mmol) in DCE (4 mL). The mixture was stirred at 60 °C for 15 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. The reaction mixture was concentrated, and then EtOAc (40 mL) and water (50 mL) were added and the layers were separated. The aqueous phase was extracted twice with EtOAc (50 mL). The combined organic layers were washed with brine (50 mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[ _4- [6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound I9, 160.0 mg), a yellow solid. MS calculated value 906.3 (MH ⁺ ); measured value 906.3 (MH ⁺ ).

Step 9: Preparation of N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]tert-butyl carbamate (compound I10)

At 0 °C, EDCI (507.8 mg, 2.65 mmol) and DIEA (0.6 mL, 3.53 mmol) were added to a solution of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound I9, 160.0 mg, 0.18 mmol) in DCM (15 mL), followed by HOBt (59.6 mg, 0.44 mmol), and the mixture was stirred at 20 °C for 15 hours. After the reaction was complete, EtOAc (50 mL) and water (80 mL) were added, and the layers were separated. The aqueous phase was extracted twice with EtOAc (50 mL). The combined organic layers were washed with brine (80 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (EtOAc = 25%-66% in PE) to give N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound I10, 130.0 mg), which is a grayish-white solid. MS: Calculated value 888.3 (MH ⁺ ); Measured value 888.3 (MH ⁺ )

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

To N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of tert-butyl carbamate (compound I10, 130.0 mg, 0.15 mmol) in DCM (1.5 mL) was mixed with TFA (0.6 mL, 8.27 mmol) and stirred at 20 °C for 1 h. After the reaction was complete, a saturated _NaHCO3 solution (40 mL) was added and the mixture was extracted three times with EtOAc (50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate I, 105.0 mg), which was a yellow solid. MS calculated value 788.3 (MH ⁺ ); measured value 788.3 (MH ⁺ ).

Intermediate J

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

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

intermediate K

(8S,14S)-8-amino-26-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 ]nonadecano-1(26),2,4,6(29),20,23(27),24-heptaen-9,15-dione

Similar to the preparation of intermediate G, the title compound was prepared by using 4-[(5M)-5-[5-bromo-4-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound K8) instead of 4-[(5M)-5-[5-bromo-1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D8).

Similar to the preparation of 4-[(5M)-5-[5-bromo-1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound D8), 4-[(5M)-5-[5-bromo-4-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (compound K8) was prepared by replacing 5-bromo-4-fluoro-1H-indol with 5-bromo-6-fluoro-1H-indol (compound D2).

intermediate L

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

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

intermediate M

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

Step 1: Preparation of [3-[5-bromo-1-ethyl-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound M1)

At 0 °C, Cs₂CO₃ (19.3 g, 59.3 mmol) and iodoethane (6.16 g, 39.53 mmol) were added to a solution of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl] _-5 -morpholino- ₃ -pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound I₄, 15 g, 19.77 mmol) in DMF (300 mL). After stirring at 20 °C for 16 hours, the reaction mixture was poured into water (200 mL) and extracted three times with EtOAc (200 mL). The combined organic layers were washed three times with brine (10 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give the residue. The residue was purified by column chromatography to give [3-[5-bromo-1-ethyl-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound M1, 14.7 g), which was a yellow oil. MS calculated value 786.3 (MH ⁺ ); measured value 786.4 (MH ⁺ ).

Step 2: Preparation of 3-[5-bromo-1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol (compound M2) and 3-[5-bromo-1-ethyl-6-fluoro-(2P)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol (compound M3)

Cesium fluoride (14.2 g, 93.41 mmol) was added to a solution of [3-[5-bromo-1-ethyl-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound M1, 14.7 g, 18.68 mmol) in DMF (160 mL). The mixture was stirred at 60 °C for 48 h. After cooling to room temperature, EtOAc (300 mL) and water (300 mL) were added to the reaction mixture, and the layers were separated. The aqueous phase was extracted three times with EtOAc (200 mL). The combined organic layers were washed four times with brine (200 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give the residue. The residues were purified by column chromatography to give 3-[5-bromo-1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol (compound M2, 6 g, fast elution), which was a colorless foam; and 3-[5-bromo-1-ethyl-6-fluoro-(2P)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop-1-ol (compound M3, 4.5 g, slow elution), which was a colorless foam. Compound M2: MS calculated value 548.2 (MH ⁺ ); measured value 548.2 (MH ⁺ ). ¹ H NMR (400MHz, methanol-d ₄ )δ＝8.41(d,J＝2.4Hz,1H),7.92(d,J＝6.8Hz,1H),7.37-7.33(m,2H),4.58(s,1H),4.05-3.98(m,2H),3.87-3.82(m,5H),3.27-3.23(m,4H),3 .15-3.13(m,1H),3.00(s,3H),2.75-2.71(m,1H),2.24-2.22(m,1H),1 .42(d,J＝6.4Hz,3H),1.22(t,J＝7.2Hz,3H),0.76(s,3H),0.76(s,3H).

X-ray crystallographic analysis of compound M2

The absolute configuration of compound M2 was confirmed by X-ray crystallography analysis of its single crystal (Figure 3).

Step 3: Preparation of 3-[1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound M4)

Potassium acetate (2.7 g, 27.35 mmol) and Pd(dppf)Cl₂ (0.8 g, 1.09 mmol) were added to a solution of 3-[5-bromo-1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-3-yl]-2,2-dimethyl-prop- ₁ -ol (compound M₂, 6 g, 10.94 mmol), bis(pinacol)diboron (4.2 g, 16.41 mmol), and toluene (60 mL). The reaction mixture was degassed by purging nitrogen for 5 min and then stirred at 90 °C for 15 h. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated under vacuum to obtain the residue. The residue was purified by column chromatography to give 3-[1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound M4, 4.5 g), which was a colorless gel. MS calculated value 596.4 (MH ⁺ ); measured value 596.4 (MH ⁺ ).

Step 4: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid methyl ester (compound M5)

Add K₃PO₄ (4.0 g, 18.89 mmol) and Pd(dtbpf)Cl to a mixture of 3-[1-ethyl-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)indol-3-yl]-2,2-dimethyl-prop-1-ol (compound M4, 4.5 g, 7.56 mmol) and (3S)-1-[(2S) _-3- ( ₄ -bromothiazol-2-yl)-2-(tert-butoxycarbonylamino)propionyl]hexahydropyridazine-3-carboxylate (intermediate C, 3.6 g, 7.56 mmol) in toluene (45 mL), 1,4-dioxane (15 mL), and water (15 mL). ₂ (492.5 mg, 0.75 mmol). The mixture was stirred at 70 °C for 12 hours under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated under vacuum to give a residue. The residue was purified by column chromatography to give methyl (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylate (compound M5, 3.8 g), as a colorless gel. MS calculated value 866.4 (MH ⁺ ); measured value 866.4 (MH ⁺ ).

Step 5: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound M6)

Me 3 SnOH (3.2 g, 17.55 mmol) was added to a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3 _- carboxylate (compound M5, 3.8 g, 4.39 mmol) in a DCE (76 mL). The mixture was stirred at 60 °C for 48 h. The reaction mixture was concentrated under vacuum to obtain a residue. EtOAc (200 mL) and water (100 mL) were added to the residue, and the layers were separated. The aqueous phase was extracted twice with EtOAc (150 mL). The combined organic layers were washed with brine (200 mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[ _4- [1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridinyl]indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound M6, 3.7 g), a brown solid. MS calculated value 852.4 (MH ⁺ ); measured value 852.4 (MH ⁺ ).

Step 6: Preparation of N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]tert-butyl carbamate (compound M7)

At 0 °C, DIEA (7.58 mL, 58.68 mmol), EDCI (8.4 g, 44.01 mmol), and HOBt (991.2 mg, 0.91 mmol) were added to a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[1-ethyl-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]indol-5-yl]thiazolyl]propionyl]hexahydropyridazine-3-carboxylic acid (compound M6, 2.5 g, 2.93 mmol) in DCM (250 mL). After stirring at 20 °C for 12 hours, the reaction mixture was poured into water (100 mL) and extracted three times with EtOAc (100 mL). The combined organic layers were washed with brine (30 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue. The residue was purified by column chromatography to obtain N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound M7, 1.2 g), which is a yellow oil. MS calculated value 834.4 (MH ⁺ ); measured value 834.4 (MH ⁺ ).

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

TFA (6.0 mL) was added to a solution of N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl] tert-butyl carbamate (compound M7, 1.2 g, 1.44 mmol) in DCM (12 mL). The mixture was stirred at 20 °C for 3 hours. After the reaction was complete, the reaction mixture was concentrated under vacuum to obtain the residue. Add saturated _NaHCO3 aqueous solution (60 mL) and extract the mixture three times with EtOAc (80 mL). Wash the combined organic layers with brine (100 mL), dry with anhydrous sodium sulfate, filter and concentrate under vacuum to give (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate M, 1 g), which is a yellow solid. MS calculated value 734.3 (MH ⁺ ); measured value 734.3 (MH ⁺ ).

intermediate N

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

Similar to the preparation of intermediate I, the title compound was prepared by using 2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)pyridine (compound N1) instead of 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-3-pyridyl]morpholine (compound I3).

Similar to the preparation of intermediate A, compound N1 was prepared by using 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (compound A1) instead of 4-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylic acid benzyl ester (intermediate A5).

Example 1

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide]

The compound was prepared according to the following scheme:

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

At 0 °C, the reaction proceeds to (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E, 150.0 mg, 0.19 mmol) in DMF (1.5 mL) was added to a solution of BOC-N-ME-VAL-OH (compound 1A, 75.0 mg, 0.32 mmol), DIEA (0.1 mL, 0.58 mmol), and HATU (120.0 mg, 0.32 mmol) in DMF (1.5 mL), and the mixture was stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was poured into water (30 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed three times with brine (30 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by preparative HPLC. The eluent was concentrated under vacuum to obtain N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [C6O-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-carbamate tert-butyl ester (compound 1B, 150.0 mg), is a yellow solid. MS calculated value 1014.5 (MH ⁺ ); measured value 1014.5 (MH ⁺ ).

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

To N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of tert-butyl carbamate (compound 1B, 100.0 mg, 0.1 mmol) in DCM (1 mL) was mixed with TFA (1.0 mL), and the mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the reaction mixture was concentrated under vacuum to obtain a residue. _A saturated aqueous solution of NaHCO3 (20 mL) was added, and the mixture was extracted three times with EtOAc (20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26} [C1, 90.0 mg]-3-methyl-2-(methylamino)butyramide (compound 1C, 90.0 mg), is a yellow solid. MS calculated value 914.4 (MH ⁺ ); measured value 914.4 (MH ⁺ ).

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

At 0°C, the (2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-2-(methylamino)butyramide (compound 1C, 90.0 mg, 0.1 mmol) in DMF (1.5 mL) was added to a solution of (S)-1-Boc-pyrrolidine-3-carboxylic acid (compound 1D, 31.9 mg, 0.15 mmol), DIEA (0.1 mL, 0.33 mmol), and HATU (57.2 mg, 0.15 mmol), and the mixture was then stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was poured into water (20 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed with brine (20 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue. This residue was purified by preparative HPLC to obtain (3S)-3-[[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 [C1, 25), 2, 5, 19, 22, 26, 23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-methyl-carbamoyl]pyrrolidine-1-carboxylic acid tert-butyl ester (compound 1E, 40.0 mg), is a yellow solid. MS calculated value 1111.5 (MH ⁺ ); measured value 1111.6 (MH ⁺ ).

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

To (3S)-3-[[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of tert-butyl pyrrolidine-1-carboxylate (compound 1E, 40.0 mg, 0.04 mmol) in DCM (1 mL) was mixed with TFA (0.3 mL) and stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was concentrated under vacuum to obtain the residue. A saturated aqueous solution _of NaHCO3 (20 mL) was added and the mixture was extracted three times with EtOAc (15 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain (3S)-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [C18-hexadecimal-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (Compound 1F, 25.0 mg), is a yellow solid. MS calculated value 1011.4 (MH ⁺ ); measured value 1011.4 (MH ⁺ ).

Step 5: (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 Preparation of 2,8-octadec-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (Example 1).

At 0 °C, (2R)-2-chloro-2-fluoro-acetic acid (23.0 mg, 0.2 mmol), (3S)-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of 1,2,5,19,22,23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (compound 1F, 25.0 mg, 0.02 mmol) and DIEA (0.1 mL, 0.77 mmol) in DMF (2 mL) was mixed with _T3P (130.0 mg, 0.2 mmol) and stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was poured into water (20 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed three times with brine (30 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue. This residue was purified by preparative HPLC to obtain (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide (Example 1, 9.4 mg), is a white solid. MS calculated value 1105.4 (MH ⁺ ); measured value 1105.8 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.52(d,J＝2.8Hz,1H),7.59-7.41(m,4H),7.01-6.70(m,1H),5.97-5.85(m, 1H),5.22-5.04(m,1H),4.68-4.63(m,1H),4.43-4.35(m,1H),4.11-3.91(m,3H) ,3.81-3.48(m,13H),3.44-3.39(m,1H),3.28-3.24(m,1H),3.23-3.11(m,6H),3 .05-2.97(m,4H),2.92-2.80(m,1H),2.66-2.55(m,1H),2.39-1.97(m,4H),1.71

-1.60(m,1H),1.46(d,J＝6.4Hz,3H),1.35-1.27(m,2H),1.18(t,J＝7.2Hz,3H),0.98-0.79(m,9H),0.67-0.57(m,2H)ppm.

Example 2

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide]

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate E) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E). Example 2 (39.0 mg) was obtained as a white solid. MS calculated value 1105.4 (MH ⁺ ); measured value 1105.6 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4 ) δ＝8.68(d,J＝7.6Hz,1H),8.57-8.49(m,1H),7.79-7.66(m,1H),7.56-7.43(m,2H),6.95-6.72(m,1H),5.83-5.61(m,1H),5.30-5.10(m,1H),4.96-4.88(m,3H). 4.82(dd,J＝11.2,3.6Hz,1H),4.52-4.36(m,1H),4.29-4.17(m,2H),4.15-3.93( m,2H),3.90-3.67(m,6H),3.66-3.55(m,2H),3.55-3.41(m,2H),3.36(d,J＝2.8H z,3H),3.34-3.32(m,1H),3.30-3.23(m,2H),3.19-3.14(m,1H),3.14-3.09(m,3 H),3.03-2.96(m,3H),2.89-2.74(m,1H),2.63-2.50(m,1H),2.45-2.21(m,3H), 2.20-1.91(m,2H),1.90-1.75(m,1H),1.72-1.57(m,1H),1.45(d,J＝6.4Hz,3H), 1.11-0.98(m,3H),0.98-0.94(m,3H),0.94-0.84(m,3H),0.54-0.34(m,3H)ppm.

Example 3

(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]-17,17,24-trimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide]

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17,24-trimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E) with (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E). Example 3 (4.7 mg) was obtained as a white solid. MS calculated value 1101.5 (MH ⁺ ); measured value 1101.6 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ=8.52-8.47(m,1H),8.15-7.98(m,1H),7.54-7.37(m,3H),6.95-6.70(m,1H),5.85-5.75(m,1H),5.35-5.07(m,2H),4 .78-4.67(m,1H),4.50-4.38(m,1H),4.25-4.14(m,1H),4.12-3.95(m,2H),3.91-3.55(m,10H),3.50-3.07(m,5H),3.00( s,3H),2.79-2.64(m,1H),2.62-2.43(m,4H),2.40-2.12(m,4H),2.10-1.84(m,3H),1.72-1.49(m,3H),1.47-1.41(m,3H ),1.39-1.32(m,2H),1.29-1.23(m,3H),0.98-0.95(m,3H),0.95-0.91(m,3H),0.88-0.84(m,3H),0.43-0.36(m,3H)ppm.

Examples 4 and 5

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 4)

(Example 4)

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 5)

(Example 5)

Example 4 was prepared according to the following scheme:

Step 1: (5S)-2-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Preparation of tert-butyl nonane-7-carboxylate (compound 4A) of octadec-1(25),2,5(28),19,22(26),23-hexen-7-yl]carbamoyl]-2-methyl-propyl]-1-oxo-2,7-diazaspiro[4.4]

HATU (94.9 mg, 0.25 mmol) and DIEA (0.1 mL, 0.37 mmol) were added to a solution of (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2, 85.0 mg, 0.25 mmol) in DMF (3 mL). After stirring at 20°C for 0.2 hours, (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D, 100.0 mg, 0.12 mmol) was added. The reaction mixture was then stirred at 20°C for another 2 hours. After the reaction was complete, the reaction mixture was directly purified by reversed-phase rapid column chromatography, and the eluent was concentrated under vacuum to remove ACN. The aqueous phase was neutralized with _NaHCO3 and extracted three times with EtOAc (10 mL). The combined organic layers were washed with brine (30 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain (5S)-2-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [C1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-1-oxo-2,7-diazaspiro[4.4]nonane-7-carboxylic acid tert-butyl ester (compound 4A, 120.0 mg), is a yellow solid. MS calculated value 1123.6 (MH ⁺ ); measured value 1123.6 (MH ⁺ ).

Step 2: Preparation of (2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexen-7-yl]-3-methyl-2-[(5R)-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]butyramide (compound 4B)

To (5S)-2-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-1-oxo-2,7-diazaspiro[4.4]nonane-7-carboxylic acid tert-butyl ester (compound 4A, 120.0 mg, 0.11 mmol) was added to a solution of TFA (1.0 mL, 12.98 mmol) in DCM (3 mL). The reaction mixture was stirred at 20 °C for 0.5 h. After the reaction was complete, the reaction mixture was concentrated under vacuum. The residue was washed with EtOAc (10 mL) and neutralized with _NaHCO3 solution. The organic phase was separated and the aqueous phase was then extracted twice with EtOAc (10 mL). The combined organic layers were washed with brine (30 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain (2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadec-1(25),2,5(28),19,22(26),23-hexen-7-yl]-3-methyl-2-[(5R)-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]butyramide (compound 4B, 120.0 mg), is a yellow solid. MS calculated value 1023.5 (MH ⁺ ); measured value 1023.5 (MH ⁺ ).

Step 3: (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Preparation of octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methylbutyramide (Example 4)

To (2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of octadecano-1(25),2,5(28),19,22(26),23-hexen-7-yl]-3-methyl-2-[(5R)-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]butyramide (compound 4B, 110.0 mg, 0.11 mmol), (2R)-2-chloro-2-fluoroacetic acid (55.0 mg, 0.49 mmol), and DIEA (0.1 mL, 0.55 mmol) in DMF (2 mL) was added to _T3P (275.0 mg, 0.43 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was complete, _H2O (30 mL) was added to the reaction mixture, and the mixture was extracted twice with EA (10 mL). The combined organic layers were washed with brine (50 mL), _dried over _Na2SO4 , filtered, and concentrated under vacuum to obtain the residue. The residue was purified by preparative HPLC to obtain (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 4, 54.4 mg), a yellow solid. MS calculated value 1117.4 (MH ⁺ ); measured value 1117.4 (MH ⁺ ). 1H NMR (400 MHz, methanol- _d4) )δ:8.68(d,J＝7.2Hz,1H),8.50(d,J＝2.8Hz,1H),7.82-7.69(m,1H),7.52 -7.45(m,2H),6.95-6.72(m,1H),5.72-5.65(m,1H),5.21-5.14(m,1H),4. 46-4.38(m,1H),4.37-4.31(m,1H),4.27-4.20(m,2H),4.16-3.97(m,2H) ,3.97-3.88(m,1H),3.88-3.72(m,4H),3.72-3.54(m,6H),3.53-3.44(m,3 H),3.35(d,J＝4.2Hz,2H),3.19-3.13(m,2H),3.00(s,3H),2.86-2.78(m, 1H),2.62-2.51(m,1H),2.34-2.19(m,3H),2.18-2.07(m,3H),2.06-1.93( m,2H),1.89-1.77(m,1H),1.69-1.60(m,1H),1.45(d,J＝6.0Hz,3H),1.37- 1.26(m,2H),1.03-0.96(m,6H),0.93-0.89(m,3H),0.46-0.40(m,3H)ppm.

Similar to the preparation of Example 4, Example 5 was prepared by using (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1) instead of (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2). Example 5 (44 mg) was obtained as a white solid. MS calculated value 1117.4 (MH ⁺ ); measured value 1117.4 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ=8.74-8.68(m,1H),8.56-8.50(m,1H),7.76-7.69(m,1H),7.62-7.56(m,1H),7.5 4-7.47(m,1H),6.96-6.73(m,1H),5.75-5.64(m,1H),5.20(dd,J＝8.4,16.4Hz,1H), 4.46-4.40(m,1H),4.35(dd,J＝4.8,11.1Hz,1H),4.31-4.19(m,2H),4.18-4.02(m,2 H),3.95(dd,J＝4.9,10.1Hz,1H),3.85-3.72(m,4H),3.71-3.61(m,3H),3.56-3.42(m ,4H),3.38(s,4H),3.27(dd,J＝5.1,10.0Hz,2H),3.21-3.14(m,1H),3.21-3.12(m,1 H),3.01(s,3H),2.90-2.79(m,1H),2.60(d,J＝14.5Hz,1H),2.33-2.20(m,3H),2.17 -2.05(m,3H),2.02-1.92(m,1H),1.90-1.78(m,1H),1.76-1.60(m,1H),1.48(d,J＝6 .1Hz,3H),1.37-1.30(m,1H),1.06-0.99(m,6H),0.95-0.87(m,3H),0.47(s,3H)ppm.

Compounds 4f1 and 4f2 were prepared according to the following scheme:

Step 1: Preparation of 1-(tert-butyl)-3-methyl-3-allylpyrrolidine-1,3-dicarboxylic acid (compound 4b).

Under a nitrogen atmosphere, at -70 °C, LDA (12 mL, 24 mmol) was added dropwise to a solution of 1-(tert-butyl)-3-methylpyrrolidine-1,3-dicarboxylate (compound 4a, 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 reaction mixture was poured into a saturated _NH₄Cl solution (100 mL) and extracted twice with EtOAc (70 mL). The combined organic layers were washed with brine (70 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by silica gel column chromatography to give 1-(tert-butyl)-3-methyl-3-allylpyrrolidine-1,3-dicarboxylate (compound 4b, 2.91 g), which was a colorless oil.

Step 2: Preparation of 1-(tert-butyl)-3-methyl-3-(2-oxoethyl)pyrrolidine-1,3-dicarboxylic acid (compound 4c)

At 0 °C, 2,6-dimethylpyridine (1.8 mL, 15.6 mmol) and K₂O₅SO₄ (0.1 g, 0.39 mmol) were added in a single batch to a mixture of 1-(tert-butyl)-3-methyl-3-allylpyrrolidine- _1,3- dicarboxylate (compound _4b , 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 (6.6 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, it was quenched with a saturated aqueous solution _{of Na₂S₂O₃} ( ₁₀₀ mL), and the reaction mixture was extracted three times with EtOAc (50 mL). The combined organic layers 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-dicarboxylate (compound 4c, 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 4d).

At 0 °C, zinc chloride (1.05 g, 7.74 mmol) was added in a single addition to a mixture of 1-(tert-butyl)-3-methyl-3-(2-oxoethyl)pyrrolidine-1,3-dicarboxylate (compound 4c, 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 poured into a saturated aqueous solution of _NH₄Cl (40 mL) at 0 °C and extracted three times with EtOAc (50 mL). The combined organic layers were washed four times with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by silica gel column chromatography to give 1-(tert-butyl)-3-methyl-3-(2-(((S)-1-(benzyloxy)-3-methyl-1-oxobutane-2-yl)amino)ethyl)pyrrolidine-1,3-dicarboxylate (compound 4d, 2.2 g), which was a yellow oil. MS calculated value 463.3 (MH ⁺ ); measured value 463.2 (MH ⁺ ).

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

DIEA (7.9 mL, 45.4 mmol) and DMAP (0.6 g, 4.54 mmol) were added in a single addition to a mixture of 1-(tert-butyl)-3-methyl-3-(2-(((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)amino)ethyl)pyrrolidine-1,3-dicarboxylate (compound 4d, 2.1 g, 4.54 mmol) in toluene (20 mL). 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 layers were washed with brine (30 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to give a residue, which was purified by reverse-phase rapid column chromatography, and the eluent was concentrated under vacuum. The residue was further separated by preparative SFC to obtain tert-butyl 7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (compound 4e 1, faster elution, 521 mg; compound 4e 2, slower elution, 525 mg). MS calculated value 453.3 (MNa ⁺ ); measured value 453.2 (MNa ⁺ ).

Step 5: Preparation of (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1).

A solution of (R)-7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (compound 4e 1, 120 mg, 0.28 mmol) in toluene (2 mL) was added with wet palladium (12 mg, 10% wt, supported on activated carbon). The mixture was degassed and purged three times with hydrogen. The reaction mixture was heated to 35 °C and stirred for 3 hours under a hydrogen atmosphere. After the reaction was complete, the solution was filtered and the filtrate was concentrated under vacuum to give (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1, 94 mg), which was a white solid. MS calculated value 363.2 (MNa ⁺ ); measured value 363.1 (MNa ⁺ ).

X-ray crystallographic analysis of compound 4f1

The absolute configuration of compound 4f1 was confirmed by X-ray crystallography of its single crystal. (Figure 1)

Step 6: Preparation of (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2).

A solution of (S)-7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (compound 4e 2, 120 mg, 0.28 mmol) in toluene (2 mL) was added with wet palladium (12 mg, 10% wt, supported on activated carbon). 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 and the filtrate was concentrated under vacuum to give (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2, 79 mg), which was a white solid. MS calculated value 363.2 (MNa ⁺ ); measured value 363.1 (MNa ⁺ ).

X-ray crystallographic analysis of compound 4f2

The absolute configuration of compound 4f2 was confirmed by X-ray crystallography of its single crystal. (Figure 2)

Example 6

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Example 6 was prepared according to the following scheme:

Step 1: 2-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] Preparation of tert-butyl undecane-9-carboxylate (compound 6K) of octadecane-1(25),2,5(28),19,22(26),23-hexen-7-yl]carbamoyl]-2-methyl-propyl]-1-oxo-2,9-diazaspiro[5.5]

Add DIEA (0.2 mL, 0.94 mmol) and HATU (106.8 mg, 0.28 mmol) to a solution of (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J, 138.0 mg, 0.37 mmol) in DMF (2 mL). After stirring at 20°C for 10 min, a solution of (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D, 150.0 mg, 0.19 mmol) in DMF (2 mL) was added to the reaction mixture, and the reaction mixture was stirred at 20°C for 50 min. After the reaction was complete, the reaction mixture was added to water (20 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed three times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which was purified by reverse-phase rapid column chromatography to give 2-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadec-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-1-oxo-2,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (compound 6K, 125.0 mg), is a yellow solid. MS calculated value 1151.6 (MH ⁺ ); measured value 1151.8 (MH ⁺ ).

Step 2: Preparation of (2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexen-7-yl]-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)butyramide (compound 6L)

To 2-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Tar-butyl 2,5,19,22,23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-1-oxo-2,9-diazaspiro[5.5]undecane-9-carboxylic acid (compound 6K, 125.0 mg, 0.11 mmol) was added to a solution of TFA (0.6 mL, 7.73 mmol) in DCM (1.5 mL). The solution was stirred at 20 °C for 0.5 h. After the reaction was complete, the reaction mixture was added to a saturated aqueous solution of _NaHCO3 (20 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed three times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain (2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26} [Octadec-1(25),2,5(28),19,22(26),23-hexen-7-yl]-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)butyramide (compound 6 L, 110.0 mg), is a yellow solid. MS calculated value 1051.5 (MH ⁺ ); measured value 1051.7 (MH ⁺ ).

Step 3: (2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Preparation of octadecano-1(25),2,5(28),19,22(26),23-hexen-7-yl]-3-methylbutyramide (Example 6)

At 0°C, the (2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of octadecano-1(25),2,5(28),19,22(26),23-hexen-7-yl]-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)butyramide (compound 6L, 110.0 mg, 0.1 mmol), (R)-2-chloro-2-fluoroacetic acid (58.8 mg, 0.52 mmol) in DMF (3 mL) was supplemented with DIEA (0.1 mL, 0.52 mmol) and _T3P (99.9 mg, 0.16 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was poured into water (40 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed three times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by preparative HPLC to obtain (2S)-2-[9-[(2R)-2-chloro-2-fluoroacetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0] ^22,26 ]Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 6, 17.7 mg), a white solid. MS calculated value 1145.5 (MH ⁺ ); measured value 1145.3 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.69(d,J＝6.4Hz,1H),8.50(d,J＝2.8Hz,1H),7.74-7.70(m,1H),7.53-7.45(m,2H),7.09-6.91(m,1H),5.70-5.63(m,1H),5.2 2-5.13(m,1H),4.97-4.89(m,3H),4.83-4.77(m,1H),4.44-4.36(m,1H),4.27-4.20(m,2H),4.17-3.89(m,4H),3.80-3.69(m,3H), 3.65-3.59(m,1H),3.52-3.41(m,4H),3.36(s,5H),3.24-3.11(m,3H),2.99(s,3H),2.87-2.78(m,1H),2.61-2.54(m,1H),2.31-2. 08(m,4H),1.97-1.80(m,6H),1.69-1.59(m,3H),1.45(d,J＝6.0Hz,3H),1.02-0.94(m,6H),0.87(d,J＝6.4Hz,3H),0.44(s,3H)ppm.

Compound 6J was prepared according to the following scheme:

Step 1: Preparation of _O1 -tert-butylO4- _methyl4 -but-3-enylpiperidine-1,4-dicarboxylate (compound 6C)

Under a nitrogen atmosphere, at -70 °C, LDA (11.3 mL, 22.61 mmol) was added to a solution of O _₁-tert-butylO_₄-methylpiperidine-1,4-dicarboxylate (compound 6A, 5.0 g, 20.55 mmol) in THF (80 mL). After stirring for 0.5 h, 4-bromobut-1-ene (compound 6B, 3.0 g, 22.61 mmol) was added to the reaction mixture, and the resulting mixture was heated to 20 °C and stirred for another 2.5 h. After the reaction was complete, it was quenched by adding a saturated NH _₄Cl solution at 0 °C, and the reaction mixture was extracted twice with EtOAc (50 mL). The combined organic layers were washed with brine, _dried over _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc = 5% to 30% in PE) to give _O₁ -tert- _butylO₄ -methyl4-but-3-enylpiperidine-1,4-dicarboxylate (compound 6C, 4.13 g), which is a yellow oil. ¹ HNMR (400MHz, chloroform-d) δ5.69-5.81(m,1H),4.91-5.06(m,2H),3.87(d,J＝14.0Hz,2H),3.71(s,3H),2.92-2.8 2(m,2H),2.11(d,J=13.6Hz,2H),2.00-1.92(m,2H),1.63-1.58(m,2H),1.45(s,9H),1.40-1.32(m,2H)ppm.

Step 2: Preparation of _O1 -tert- _butylO4 -methyl4-(3-oxopropyl)piperidine-1,4-dicarboxylate (compound 6D)

At 0 °C, 2,6-dimethylpyridine (1.5 mL, 12.98 mmol) and potassium (VI) osmium tetroxide dihydrate (119.5 mg, 0.32 mmol) were added to a mixture of O _₁ -tert-butylO₄-methyl-4-but-3-enylpiperidine-1,4-dicarboxylate (compound 6C, 1.9 g, 6.49 mmol) in 1,4-dioxane (40 mL) and water (4 mL). After stirring at 0 °C for 15 min, sodium periodate (5.6 g, 25.96 mmol) was added to the reaction mixture at 0 °C, and the resulting mixture was warmed to 20 °C and stirred for 3 h. After the reaction was complete, the reaction mixture was extracted three times with EtOAc (20 mL) and washed with a saturated aqueous solution of Na _₂ SO_₃ (20 mL). The combined organic layers were washed with brine (20 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to give _O₁ -tert- _butylO₄ -methyl 4-(3-oxopropyl)piperidine-1,4-dicarboxylate (compound 6D, 2.0 g), which was a yellow oil. MS calculated value 200.2 (M-Boc+ ^H⁺ ), measured value 200.1 (M-Boc+ ^H⁺ ).

Step 3: Preparation of _O1 -tert- _butylO4 -methyl4-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]piperidine-1,4-dicarboxylate (compound 6F)

Zinc chloride (500.7 mg, 3.67 mmol) was added to a mixture of O _₁-tert-butylO_₄-methyl 4-(3-oxopropyl)piperidine-1,4-dicarboxylate (compound 6D, 1.0 g, 3.34 mmol) and Boc-N-Me-Val-OH (770.5 mg, 3.67 mmol) in methanol (20 mL) at 0 °C. After stirring at 0 °C for 0.5 h, sodium cyanoborohydride (272.8 mg, 4.34 mmol) was added to the reaction mixture at 0 °C, and the resulting mixture was warmed to 20 °C and stirred for another 1 h. After the reaction was complete, it was quenched with a saturated aqueous solution of NH _₄Cl (20 mL), and the reaction mixture was extracted three times with EtOAc (20 mL). The combined organic layers were washed with brine (20 mL), dried _over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc = 10% to 30% in PE) to give _O₁ -tert- _butylO₄ -methyl4-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]piperidine-1,4-dicarboxylate (compound 6F, 1.1 g), as a yellow oil. MS calculated value 457.3 (MH ⁺ ); measured value 457.2 (MH ⁺ ).

Step 4: Preparation of lithium; 1-tert-butoxycarbonyl-4-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]piperidine-4-carboxylate (compound 6G)

Lithium hydroxide monohydrate (367.6 mg, 8.76 mmol) was added to a mixture of O_₁-tert-butylO_₄-methyl-4-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]piperidine-1,4-dicarboxylate (compound 6F, 400.0 mg, 0.88 mmol) in methanol (6 mL), THF (0.6 mL), and water (0.6 mL). The resulting mixture was heated to 60 °C and stirred for 12 hours. After the reaction was complete, the reaction mixture was cooled to 20 °C and concentrated under vacuum to give lithium; 1-tert-butoxycarbonyl-4-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]piperidine-4-carboxylate (compound 6G, 392.0 mg), which is a white solid. MS calculated value 443.3 (M-Li+H ⁺ ); measured value 443.2 (M-Li+H ⁺ ).

Step 5: Preparation of tert-butyl 2-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-1-oxo-2,9-diazaspiro[5.5]undecane-9-carboxylate (compound 6H)

At 0 °C, DIEA (0.5 mL, 2.62 mmol) and COMU (673.7 mg, 1.57 mmol) were added to a mixture of lithium; 1-tert-butoxycarbonyl-4-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]piperidine-4-carboxylate (392.0 mg, 0.87 mmol) in DMF (8 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was poured into water (80 mL) and extracted three times with EtOAc (20 mL). The combined organic layers were washed three times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by column chromatography (EtOAc = 5% to 25% in PE) to give tert-butyl 2-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-1-oxo-2,9-diazaspiro[5.5]undecane-9-carboxylate (compound 6H, 200.0 mg), a yellow oil. MS calculated value 425.3 (MH ⁺ ); measured value 425.3 (MH ⁺ ).

Step 6: Preparation of (2S)-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)butyric acid (compound 6I)

Trifluoroacetic acid (1.0 mL, 12.98 mmol) was added to a solution of tert-butyl 2-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-1-oxo-2,9-diazaspiro[5.5]undecane-9-carboxylic acid (tert-butyl ester, compound 6H, 200.0 mg, 0.47 mmol) in DCM (1.5 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the mixture was concentrated under vacuum to give (2S)-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)butyric acid (compound 6I, 180.0 mg, TFA salt), which was a colorless oil. MS calculated value 269.2 (MH ⁺ ); measured value 269.2 (MH ⁺ ).

Step 7: Preparation of (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J)

Sodium carbonate (99.8 mg, 0.94 mmol) and di-tert-butyl dicarbonate (123.3 mg, 0.56 mmol) were added to a solution of (2S)-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)butyric acid (compound 6I, 180.0 mg, 0.47 mmol, TFA salt) in THF (3 mL) and water (3 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the mixture was concentrated under vacuum to obtain a residue, which was purified by reverse-phase rapid column chromatography to give (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J, 400.0 mg), which was a grayish-white solid. MS calculated value 367.2 (MH ^- ); measured value 367.2 (MH ^- ).

Example 8

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^{23 ,27]} [26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

Similar to the preparation in Example 1, this was achieved by using (8S,14S)-8-amino-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadecano-1(25), ^2,5 (28), ^19,22 (26),23-hexaeno-8,14-dione (intermediate E) with 29-carbon-1(26),2,4,6(29),20,23(27),24-heptaeno-9,15-dione (intermediate G). Example 8 (21.6 mg) was obtained as a white solid. MS calculated value 1114.5 (MH ⁺ ); measured value 1114.8 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ＝8.43(d,J＝2.4Hz,1H),7.62(dd,J＝10.8,7.2Hz,1H),7.55-7.53(m,2 H),7.29(s,1H),7.26-7.20(m,1H),6.96-6.90(m,1H),6.65-6.55(m,1H ),5.65-5.49(m,1H),4.97-4.68(m,2H),4.67-4.62(m,1H),4.46-4.38( m,1H),4.20-4.12(m,1H),4.00-3.87(m,1H),3.82-3.61(m,6H),3.59-3. 43(m,5H),3.38-3.32(m,2H),3.25-3.11(m,1H),3.04(s,2H),2.99-2.9 6(m,1H),2.93(s,4H),2.90-2.65(m,3H),2.63-2.47(m,2H),2.39-2.11( m,3H),2.10-1.95(m,2H),1.89-1.79(m,1H),1.58-1.27(m,6H),1.03-0 .92(m,6H),0.91-0.84(m,1H),0.83-0.75(m,3H),0.47-0.32(m,3H)ppm.

Example 9

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]-17,17,24-trimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [Octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide]

Similar to the preparation in Example 1, this was achieved by using 1-tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17,24-trimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate F) replaces (S)-1-Boc-pyrrolidine-3-carboxylic acid (compound 1D) and (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E). Example 9 (2.7 mg) was obtained as a white solid. MS calculated value 1133.5 (MH ⁺ ); measured value 1133.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.49-8.36(m,1H),8.03(d,J＝13.6Hz,1H),7.58-7.39(m,3H),7.12-6.94(m,1H),5.90-5.7 7(m,1H),5.26-5.11(m,1H),4.73-4.61(m,1H),4.52-4.31(m,2H),4.21(m,1H),4.14-3.84(m, 4H),3.75-3.49(m,3H),3.26-3.07(m,10H),3.00(s,4H),2.52(m,6H),2.15(m,8H),1.97-1.82 (m,1H),1.71-1.50(m,2H),1.45(m,3H),1.36-1.24(m,3H),1.02-0.87(m,9H),0.39(s,3H)ppm.

Example 10

1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26] [Octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide]

Similar to the preparation in Example 1, this was achieved by using 1-tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D) replaces (S)-1-Boc-pyrrolidine-3-carboxylic acid (compound 1D) and (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E). Example 10 (17.5 mg) was obtained as a yellow solid. MS calculated value 1137.5 (MH ⁺ ); measured value 1137.5 (MH ⁺ ), ^1H NMR (400 MHz, methanol- _d4) )δ=8.74-8.68(m,1H),8.54-8.51(m,1H),7.74-7.70(m,1H),7.57-7.47(m,2H),7.15-6.97(m,1H),5.75-5.64(m,1H),5.26-5.12( m,1H),4.83-4.73(m,1H),4.51-4.32(m,2H),4.29-4.21(m,2H),4.20-3.89(m,3H),3.88-3.62(m,5H),3.60-3.43(m,4H),3.40-3.3 6(m,4H),3.26-3.22(m,4H),3.04-3.00(m,4H),2.89-2.81(m,1H),2.64-2.56(m,1H),2.39-2.15(m,6H),1.98-1.95(m,1H),1.89- 1.78(m,1H),1.73-1.59(m,1H),1.50-1.43(m,4H),1.38-1.28(m,1H),1.04-0.97(m,6H),0.95-0.88(m,3H),0.51-0.41(m,3H)ppm.

Example 11

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.5]nonane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation of Example 6, the title compound was prepared by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.5]nonane-6-yl)-3-methyl-butyric acid (compound 11J) instead of (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J). Example 11 (6.6 mg) was obtained as a white solid. MS calculated value 1117.4 (MH ⁺ ); measured value 1117.5 (MH ⁺ ). ¹ H NMR (400MHz, methanol-d ₄ )δ＝8.68(d,J＝7.3Hz,1H),8.50(d,J＝2.8Hz,1H),7.72(dd,J＝2.4,15.6Hz,1H),7. 50-7.45(m,2H),6.79-6.60(m,1H),5.72-5.64(m,1H),5.21-5.10(m,3H),4.44(br s,2H),4.21(br d,J＝5.6Hz,3H),4.05-3.86(m,3H),3.79(br d,J＝11.2Hz,1H),3.74-3.61(m,4H),3.57-3.40(m,8H),3.35(br s,2H),3.15(br d,J＝14.8Hz,3H),3.00(s,3H),2.83(br t,J＝11.6Hz,1H),2.62-2.50(m,1H),2.20(br s,3H),1.99-1.94(m,1H),1.83(br s,3H),1.64(br dd,J＝4.0,12.0Hz,1H),1.44(d,J＝6.0Hz,3H),1.00(br dd,J＝2.8,6.4Hz,3H),0.97(s,3H),0.89(br d,J＝6.4Hz,3H),0.44(s,3H)ppm.

Compound 11J was prepared according to the following scheme:

Step 1: Preparation of tert-butyl 3-but-3-enyl-3-cyano-azacyclobutane-1-carboxylate (compound 11B)

Under a nitrogen atmosphere, lithium bis(trimethylsilyl)amide (30.1 mL, 30.18 mmol) was added dropwise to a solution of tert-butyl 3-cyanozycyclobutane-1-carboxylate (compound 11A, 5.0 g, 27.44 mmol) in THF (50 mL) cooled to -70 °C. After stirring for 0.5 h, 4-bromo-1-butene (4.4 g, 32.93 mmol) was slowly added to the reaction mixture at -70 °C. The mixture was heated to 20 °C and stirred for 3 h. After the reaction was complete, it was quenched with saturated _NH₄Cl solution (200 mL) and the reaction mixture was extracted three times with EtOAc (50 mL). The combined organic layers were washed with brine (100 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to obtain a residue. This residue was purified by column chromatography (EtOAc = 5% to 30% in PE) to give tert-butyl 3-but-3-enyl-3-cyano-azacyclobutane-1-carboxylate (compound 11B, 5.4 g), which was a yellow oil. MS calculated value 237.2 ( ^MH⁺ ); measured value _181.6 ( _{MC₄H₈⁺H⁺} ⁾ .

Step 2: Preparation of 3-but-3-enyl-1-tert-butoxycarbonyl-azacyclobutane-3-carboxylic acid (compound 11C)

Potassium hydroxide (5.1 g, 91.41 mmol) was added once to a mixture of tert-butyl 3-but-3-enyl-3-cyano-azacyclobutane-1-carboxylic acid (compound 11B, 5.4 g, 22.85 mmol) in ethanol (60 mL) and water (60 mL). The mixture was heated at 80 °C for 16 h. After the reaction was complete, the pH of the reaction mixture was adjusted to pH 3 with 1 N HCl aqueous solution and extracted three times with EtOAc (50 mL). The combined organic layers were washed with brine (100 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to give 3-but-3-enyl-1-tert-butoxycarbonyl-azacyclobutane-3-carboxylic acid (compound 11C, 6.31 g) as a yellow oil. MS calculated value 256.2 ( ^MH⁺ ); measured value _200.3 ( _{MC₄H₈⁺H⁺} ⁾ .

Step 3: Preparation of _O1 -tert-butylO3-methyl3-but- ₃ -enylazonylbutane-1,3-dicarboxylate (compound 11D)

Potassium carbonate (10.2 g, 74.15 mmol) and methyl iodoformate (7.0 g, 49.43 mmol) were added in a single addition to a mixture of 3-but-3-enyl-1-tert-butoxycarbonyl-azacyclobutane-3-carboxylic acid (compound 11C, 6.3 g, 24.72 mmol) in DMF (60 mL). The mixture was stirred at 20 °C for ₂ hours. After the reaction was complete, the mixture was poured into water (150 mL) and extracted three times with EtOAc (70 mL). The combined organic layers were washed three times with brine (70 mL), dried over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by column chromatography (EtOAc = 5% to 30% in PE) to give _O₁ -tert-butylO₃-methyl3-but- ₃ -enylazacyclobutane-1,3-dicarboxylic acid ester (compound 11D, 3.76 g), which is a yellow oil. MS calculated value 270.2 (MH ⁺ ); measured value 214.3 ( _MC4H8 + _H ⁺ ).

Step 4: Preparation of _O1 -tert- _butylO3 -methyl 3-(3-oxopropyl)azacyclobutane-1,3-dicarboxylate (compound 11E)

At 0 °C, 2,6-dimethylpyridine (3.2 mL, 27.92 mmol) and potassium (VI) osmium tetroxide dihydrate (0.3 g, 0.7 mmol) were added in a single batch to a mixture of O_₁ -tert-butylO₃-methyl-3-but-3-enylazetane-1,3-dicarboxylate (compound 11D, 3.7 g, 13.96 mmol) in 1,4-dioxane (100 mL) and water (10 mL). After stirring at 0 °C for 15 min, sodium periodate (11.9 g, 55.84 mmol) was added to the reaction mixture, and the resulting mixture was warmed to 20 °C and stirred for another 3 h. After the reaction was complete, the reaction was quenched by adding saturated aqueous Na _₂ SO_₃ solution (200 mL), and the reaction mixture was extracted three times with EtOAc (100 mL). The combined organic layers were washed with brine (100 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to give _O₁ -tert- _butylO₃ -methyl 3-(3-oxopropyl)azacyclobutane-1,3-dicarboxylate (compound 11E, 3.8 g), which was a yellow oil. MS calculated value 272.2 ( ^MH⁺ ); measured value _216.3 ( _{MC₄H₈⁺H⁺} ⁾ .

Step 5: Preparation of _O1 -tert-butylO3-methyl3-[ ₃ -[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]azacyclobutane-1,3-dicarboxylate (compound 11F)

Zinc chloride (1.4 g, 10.5 mmol) was added in a single step to a mixture of L-valine tert-butyl ester hydrogen chloride (1.8 g, 10.5 mmol) and _O1 - _tert -butylO3-methyl 3-(3-oxopropyl)azacyclobutane-1,3-dicarboxylate (compound 11E, 1.9 g, 7.0 mmol) in methanol (20 mL). After stirring at 20 °C for 0.5 h, sodium cyanoborohydride (0.9 g, 14.01 mmol) was added to the reaction mixture at 0 °C, and the resulting mixture was stirred at 20 °C for another 1 h. After the reaction was complete, it was quenched by adding saturated _NH4Cl aqueous solution (100 mL), and the reaction mixture was extracted three times with EtOAc (50 mL). The combined organic layers were washed with brine (50 mL), dried _over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc = 10% to 30% in PE) to give _O₁ -tert-butylO₃-methyl _3- [3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]azacyclobutane-1,3-dicarboxylate (compound 11F, 1.2 g), as a yellow oil. MS calculated value 429.3 (MH ⁺ ); measured value 429.4 (MH ⁺ ).

Step 6: Preparation of lithium; 1-tert-butoxycarbonyl-3-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]azacyclobutane-3-carboxylate (compound 11G)

Lithium hydroxide monohydrate (1.2 g, 28.0 mmol) was added to a mixture of O ₁ -tert-butylO ₃ -methyl 3-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]azacyclobutane-1,3-dicarboxylate (compound 11F, 1.2 g, 2.8 mmol) in methanol (30 mL), water (3 mL), and THF (3 mL). The mixture was then heated to 60 °C and stirred for 12 hours. After the reaction was complete, the reaction mixture was concentrated under vacuum to give lithium; 1-tert-butoxycarbonyl-3-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]azacyclobutane-3-carboxylate (compound 11G, 2.3 g), as a yellow solid. MS calculated value 421.8 (MH ⁺ ); measured value 415.2 (M-Li+H ⁺ ).

Step 7: Preparation of tert-butyl 6-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-5-oxo-2,6-diazaspiro[3.5]nonane-2-carboxylate (compound 11H)

At 0 °C, DIEA (2.1 mL, 12.06 mmol) and COMU (2.0 g, 4.82 mmol) were added in a single step to a solution of lithium; 1-tert-butoxycarbonyl-3-[3-[[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]amino]propyl]azacyclobutane-3-carboxylate (compound 11 G, 2.0 g, 2.41 mmol) in DMF (30 mL). The reaction mixture was then heated to 20 °C and stirred for 1 h. After the reaction was complete, the mixture was poured into water (100 mL) and extracted three times with EtOAc (50 mL). The combined organic layers were washed three times with brine (50 mL), dried _over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc = 20% to 30% in PE) to give tert-butyl 6-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-5-oxo-2,6-diazaspiro[3.5]nonane-2-carboxylate (compound 11H, 512.0 mg), as a yellow solid. MS calculated value 397.3 (MH ⁺ ); measured value 419.4 (MNa ⁺ ).

Step 8: Preparation of (2S)-3-methyl-2-(5-oxo-2,6-diazaspiro[3.5]nonane-6-yl)butyric acid (compound 11I)

TFA (3.0 mL, 38.94 mmol) was added to a solution of tert-butyl 6-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-5-oxo-2,6-diazaspiro[3.5]nonane-2-carboxylic acid (compound 11H, 512.0 mg, 1.29 mmol) in DCM (3 mL). The mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the mixture was concentrated under vacuum to give (2S)-3-methyl-2-(5-oxo-2,6-diazaspiro[3.5]nonane-6-yl)butyric acid (compound 11I, 301.0 mg), which was a yellow oily substance as a TFA salt. MS calculated value 241.2 (MH ⁺ ); measured value 241.5 (MH ⁺ ).

Step 9: Preparation of (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.5]nonane-6-yl)-3-methyl-butyric acid (compound 11J)

A mixture of (2S)-3-methyl-2-(5-oxo-2,6-diazaspiro[3.5]nonane-6-yl)butyric acid (compound 11I, 301.0 mg, 1.25 mmol) in THF (12 mL) was added in a single step with di-tert-butyl dicarbonate (326.9 mg, 1.5 mmol) and sodium carbonate (661.6 mg, 6.24 mmol), and the mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified by preparative HPLC to give (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.5]nonane-6-yl)-3-methyl-butyric acid (compound 11J, 303.0 mg), which was a colorless gel. MS calculated value 341.2 (MH ⁺ ); measured value 363.2 (MNa ⁺ ).

Example 13

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

The compound was prepared according to the following scheme:

Step 1: Preparation of ethyl 3-hydroxy-3-methyl-2-methylene-butyrate (compound 13C)

DIBAL-H (152.9 mL, 152.91 mmol) was added dropwise to a solution of NMO (23.8 g, 203.87 mmol) in THF (500 mL) at 0 °C. After stirring at 0 °C for 30 min, ethyl propionate-2-acetylacetate (compound 13A, 10 g, 101.94 mmol) was added dropwise to the reaction mixture at 0 °C. After stirring at 0 °C for another 1 h, acetone (compound 13B, 5.9 g, 101.94 mmol) and boron trifluoride diethyl ether (12.9 mL, 101.94 mmol) were added to the reaction mixture, and the resulting mixture was stirred at room temperature for 18 h. The reaction was post-treated by adding EA (100 mL). The organic layer was washed with HCl (1 N, 37.5 mL), saturated _NaHCO3 (10 mL), and water ( ₁₀ mL), and then dried over _Na2SO4 and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 10:1) to give ethyl 3-hydroxy-3-methyl-2-methylene-butyrate (compound 13C, 4.8 g), which was a colorless oil. ^¹H NMR (400 MHz, chloroform-d) δ = 6.14 (s, 1H), 5.83–5.70 (m, 1H), 4.23 (q, J = 7.3 Hz, 2H), 3.93 (br s, 1H), 1.49–1.37 (m, 6H), 1.31 (t, J = 7.1 Hz, 3H) ppm.

Step 2: Preparation of ethyl 2-(bromomethyl)-3-methyl-but-2-enoate (compound 13D)

At 0 °C, a solution of phosphorus tribromide (4.9 g, 18.21 mmol) was added dropwise to a solution of ethyl 3-hydroxy-3-methyl-2-methylene-butyrate (compound 13C, 4.8 g, 30.34 mmol) in 100 _mL of THF. After stirring at 15 °C for 2.5 hours, the reaction mixture was poured into ice water at 0 °C, and the aqueous layer was extracted with EA (10 mL). The organic layer was washed with brine, dried over _Na₂SO₄ , and concentrated to give a crude product, which was purified by silica gel chromatography (petroleum ether/EtOAc = 20/1 to 10/1) to give ethyl 2-(bromomethyl)-3-methyl-but-2-enoate (compound 13D, 5.3 g), which was a colorless oil. ^1H NMR (400MHz, chloroform-d) δ＝4.30(s,2H),4.24(d,J＝7.2Hz,2H),2.15(s,3H),1.97(s,3H),1.32(t,J＝7.2Hz,3H)ppm.

Step 3: Preparation of 3-(2-ethoxycarbonyl-3-methyl-but-2-enoxy)azacyclobutane-1-carboxylic acid benzyl ester (compound 13F)

At 0 °C, ethyl 2-(bromomethyl)-3-methyl-but-2-enoate (compound 13D, 1.0 g, 4.52 mmol) was added to a mixture of 3-hydroxyazacyclobutane-1-carboxylate (compound 13E, 625.0 mg, 3.02 mmol), tetrabutylammonium iodide (116.0 mg, 0.31 mmol), and sodium hydroxide (267.8 mg, 6.7 mmol) in water (2 mL) and THF (4 mL). After stirring at 10 °C for 16 hours, the reaction mixture was diluted with water (5 mL) and the resulting mixture was extracted twice with EA (10 mL). The combined organic layers were washed with brine (10 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (petroleum ether/EtOAc = 20/1 to 10/1) to give benzyl 3-(2-ethoxycarbonyl-3-methyl-but-2-enoxy)azacyclobutane-1-carboxylate (compound 13F, 590.0 mg), a colorless oil. MS calculated value 348.1 (MH ⁺ ); measured value 348.1 (MH ⁺ ).

Step 4: Preparation of tert-butyl 3-(2-ethoxycarbonyl-3-methyl-butoxy)azacyclobutane-1-carboxylate (compound 13G)

Pd/ _C (240.0 mg, 0.26 mmol) was added to a solution of 3-(2-ethoxycarbonyl-3-methyl-but-2-enoxy)azacyclobutane-1-carboxylic acid benzyl ester (compound 13F, 950.0 mg, 2.73 mmol) in methanol (25 mL) at 15 °C under _N2 atmosphere. The reaction mixture was purged with _H2 and stirred at 50 °C for 2 h under H2 atmosphere. The reaction mixture was filtered and the filter cake was washed with methanol (30 mL). The combined filtrates were concentrated under vacuum and the resulting residue was purified by silica gel chromatography (petroleum ether/EtOAc = 10/1 to 5/1) to give tert-butyl 3-(2-ethoxycarbonyl-3-methyl-butoxy)azacyclobutane-1-carboxylic acid (compound 13G, 700.0 mg), which is a colorless oil. ^1H NMR (400MHz, chloroform-d) δ＝4.25-4.11(m,3H),4.08-3.99(m,2H),3.80(dd,J＝4.3,9.4Hz,2H),3.61-3.46(m,2H),2.45(ddd,J＝4.9,7.3,9.5Hz,1H),1.97-1.86(m,1H),1.43(s,9H),1.31-1.24(m,3H),0.99-0.90(m,6H)ppm.

Step 5: Preparation of 2-[(1-tert-butoxycarbonylazonylbutane-3-yl)oxymethyl]-3-methylbutyric acid (compound 13H)

A mixture of tert-butyl 3-(2-ethoxycarbonyl-3-methyl-butoxy)azacyclobutane-1-carboxylate (compound 13G, 350.0 mg, 1.11 mmol) in methanol (6 mL) was added to an aqueous solution of NaOH (1 N, 6.0 mL, 6.0 mmol). After stirring at 50 °C for 16 hours, the reaction mixture was concentrated under vacuum. The resulting residue was diluted with water (10 mL), and the pH of the reaction mixture was adjusted to pH 5 with 1 N HCl aqueous solution. The reaction mixture was then extracted twice with EA (30 mL). The combined organic layers were washed with brine (50 mL), dried ( _Na₂SO₄ ) _, and concentrated under vacuum to give the crude product. The crude product was purified by silica gel chromatography (petroleum ether/EtOAc = 5/1 to 3/1) to obtain 2-[(1-tert-butoxycarbonylazine-3-yl)oxymethyl]-3-methyl-butyric acid (compound 13H, 200.0 mg), which is a colorless oil. ^1H NMR (400MHz, chloroform-d) δ＝4.22(tt,J＝4.3,6.4Hz,1H),4.10-4.03(m,2H),3.83(dd,J＝4.3,9.7Hz,2H),3.63-3.48(m,2H),2.50(ddd,J＝4.8,7.1,9.2Hz,1H),1.99(qd,J＝6.8,13.7Hz,1H),1.44(s,9H),0.99(dd,J＝2.9,6.8Hz,6H)ppm.

Step 6: 3-[(2R)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [C18-hexadecimal-1(25),2,5(28),19,22(26),23-hexen-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-carboxylic acid tert-butyl ester (compound 13I) and 3-[(2S)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] Preparation of tert-butyl 1-(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-carboxylate (compound 13J)

At 0 °C, the reaction proceeds to (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of octadecano-1 (25), 2, 5 (28), 19, 22 (26), 23-hexaden-8, 14-dione (intermediate D, 225.0 mg, 0.28 mmol) and 2-[(1-tert-butoxycarbonylazyrobutane-3-yl)oxymethyl]-3-methylbutyric acid (compound 13H, 104.9 mg, 0.37 mmol) in DMF (2.5 mL) was supplemented with DIEA (0.2 mL, 0.84 mmol) and HATU (213.6 mg, 0.56 mmol). After stirring at 20 °C for 1 h, the reaction mixture was diluted with water (3 mL) and extracted twice with EtOAc (8 mL). The combined organic layers were washed with brine (10 mL), _dried over _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by preparative HPLC to obtain 3-[(2R)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-carboxylic acid tert-butyl ester (compound 13I, fast elution, 45.0 mg), which is a pale yellow solid; and 3-[(2S)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0] ^[22,26 ] Octadectocarbon-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-carboxylic acid tert-butyl ester (compound 13J, slow elution, 100.0 mg), is a pale yellow solid. MS calculated value 1070.6 (MH ⁺ ); measured value 1070.6 (MH ⁺ ).

Step 7: Preparation of (2R)-2-(azacyclobutane-3-yloxymethyl)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26} ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butanamide (compound 13K)

At 0 °C, 3-[(2R)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 A solution of tert-butyl 1-(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-carboxylate (compound 13I, 45.0 mg, 0.04 mmol) in DCM (0.5 mL) was mixed with TFA (0.2 mL), and the mixture was stirred at 15 °C for 0.5 h. The reaction mixture was concentrated under vacuum to give (2R)-2-(azacyclobutane-3-yloxymethyl)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide; 2,2,2-trifluoroacetic acid (compound 13K, 40.0 mg), which is a yellow oil. MS calculated value 970.4 (MH ⁺ ); measured value 970.4 (MH ⁺ ).

Step 8: (2R)-2-[[1-[(2R)-2-chloro-2-fluoro-acetyl]azacyclobutane-3-yl]oxymethyl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Preparation of octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methylbutyramide (Example 13)

At 0 °C, the (2R)-2-(azacyclobutane-3-yloxymethyl)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methylbutyramide; 2,2,2-trifluoroacetic acid (compound 13K, 40.0 mg, 0.04 mmol) in a solution of DMF (0.2 mL) with added DIEA (0.1 mL, 0.74 mmol), _T3P (469.6 mg, 0.74 mmol) and (2R)-2-chloro-2-fluoroacetic acid (compound 13L, 83.0 mg, 0.74 mmol). After stirring at 20°C for 1 h, the reaction mixture was concentrated under vacuum, and the resulting residue was purified by preparative HPLC to obtain (2R)-2-[[1-[(2R)-2-chloro-2-fluoro-acetyl]azacyclobutane-3-yl]oxymethyl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 13, 22.6 mg), which is a white solid. MS calculated value 1064.5 (MH ⁺ ); measured value 1064.5 (MH ⁺ ). ^1H NMR (400MHz, chloroform-d) δ＝8.65(br d, J＝6.8Hz, 2H), 7.64(dd, J＝1.9, 4.9Hz, 1H), 7.26-7.20(m, 1H), 7.14(br d, J＝11.9Hz, 1H), 6.77-6.67(m, 1H), 6.41-6.22(m, 1H), 5.89(br t, J＝8.1Hz, 1H), 4.95(br dd, J＝9.0, 16.5Hz, 1H), 4.65-4.48(m, 3H), 4.46-4.21(m, 6H), 4.17-4.09(m, 1H), 3.86(br t, J＝8.1Hz, 1H), 4.46-4.21(m, 6H), 4.17-4.09(m, 1H), 3.86(br t, J＝9.0, 16.5Hz, 1H), 3.86(br t, J＝1.9, 1H), 4.46-4.21(m, 6H), 4.17-4.09(m ...46-4.21(m, 6H), 4.4 d,J＝11.1Hz,2H),3.74-3.69(m,3H),3.60-3.55(m,2H),3.52-3.45(m,2H),3.36(s,3 H),3.15-3.09(m,2H),2.94-2.87(m,3H),2.75-2.67(m,1H),2.47-2.34(m,1H),2.30- 2.21(m,2H),2.04-1.94(m,4H),1.88-1.79(m,1H),1.70-1.54(m,2H),1.50(d,J＝6.1 Hz,3H),1.30-1.20(m,1H),1.02(dd,J=4.6,6.3Hz,6H),0.97(s,3H),0.45(s,3H)ppm.

Example 14

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

Similar to the preparation in Example 13, 3-[(2S)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26]} was used. [Octadecano-1(25),2,5(28),19,22(26),23-hexen-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-tert-butyl ester (compound 13J) replacing 3-[(2R)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26]} The title compound was prepared by reacting 1,2,5,19,22,23-hexene-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-carboxylic acid tert-butyl ester (compound 13I). Example 14 (38.0 mg) was obtained as a white solid. MS calculated value 1064.5 (MH ⁺ ); measured value 1064.5 (MH ⁺ ). ¹ H NMR (400MHz, chloroform-d) δ = 8.72 (brs, 1H), 8.66 (d, J = 7.4Hz, 1H), 7.65 (s, 1H), 7.14 (d, J = 12.0Hz, 1H), 6.80-6.72 (m, 1H), 6.3 9-6.23(m,1H),5.97-5.86(m,1H),5.01-4.83(m,2H),4.60-4.49(m,5H),4.36-4.22(m,6H),4.05-3.93(m,2H),3.87(br d,J＝10.4Hz,1H),3.73-3.68(m,2H),3.64-3.53(m,4H),3.45(s,1H),3.36(s,3H),3.20-3.00(m,4H),2.93(br s,3H),2.73(br t,J＝13.2Hz,1H),2.43(br d,J＝14.1Hz,1H),2.30-2.20(m,2H),2.04-1.96(m,2H),1.85-1.78(m,1H),1.69-1.62(m,1H),1.51(d,J＝6.1Hz,3H),1.06(br dd,J＝3.0,6.6Hz,3H),1.03-1.00(m,3H),0.96(br s,3H),0.46(s,3H)ppm.

Example 15

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation of Example 6, the title compound was prepared by using (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 15F) instead of (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J). Example 15 (56.8 mg) was obtained as a white solid. MS calculated value 1131.5 (MH ⁺ ); measured value 1131.5 (MH ⁺ ). ^¹H NMR (400MHz, methanol- _d⁴ ) δ＝8.86(d,J＝7.2Hz,1H),8.68(d,J＝7.6Hz,1H),8.50(d,J＝2.8Hz,1H),7.71(s,1H),7.50-7.46(m,1H),7.13-6.92(m,1H),5.68(br) s,1H),5.20-5.13(m,1H),4.41(d,J＝14.1Hz,1H),4.36-4.21(m,4H),4.20-3.85(m,4H),3.82-3.68(m,3H),3 .61(d,J＝7.3Hz,4H),3.46(d,J＝14.4Hz,4H),3.36-3.35(m,2H),3.24-3.08(m,4H),3.00(s,3H),2.90-2.78(m ,1H),2.57(d,J＝14.4Hz,1H),2.28-2.17(m,2H),2.08(s,2H),1.96(d,J＝12.8Hz,2H),1.88-1.77(m,2H),1.7 6-1.52(m,4H),1.45(d,J＝6.0Hz,3H),1.00(d,J＝6.4Hz,3H),0.97(s,3H),0.92-0.84(m,3H),0.44(s,3H)ppm.

Compound 15F was prepared according to the following scheme:

Step 1: Preparation of _O1 -tert-butylO4- _methyl4 -allylpiperidine-1,4-dicarboxylate (compound 15B)

Under a nitrogen atmosphere, at -70 °C, LDA (22.6 mL, 45.21 mmol) was added dropwise to a solution of O _₁-tert-butylO_₄-methylpiperidine-1,4-dicarboxylate (compound 15A, 10.0 g, 41.1 mmol) in THF (100 mL). After stirring for 0.5 h, allyl bromide (5.5 g, 45.21 mmol) was added to the reaction mixture at -70 °C, and the reaction mixture was then allowed to rise to 15 °C and stirred for 2 h. After the reaction was complete, it was quenched with a saturated aqueous solution of NH _₄Cl (300 mL), and the reaction mixture was extracted three times with EtOAc (100 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous Na_₂SO_₄, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (EtOAc = 5% to 30% in PE) to give _O1 -tert- _butylO4 -methyl4-allylpiperidine-1,4-dicarboxylate (compound 15B, 10.8 g), which is a yellow oil. MS calculated value 284.2 (MH ⁺ ); measured value 228.2 ( _MC4H8 + _H ⁺ ).

Step 2: Preparation of _O1 -tert- _butylO4 -methyl4-(2-oxoethyl)piperidine-1,4-dicarboxylate (compound 15C)

At 0 °C, 2,6-dimethylpyridine (4.9 mL, 42.35 mmol) and potassium (VI) osmium tetroxide dihydrate (0.4 g, 1.06 mmol) were added in a single batch to a mixture of O_₁ -tert-butylO₄-methyl-4-allylpiperidine-1,4-dicarboxylate (compound 15B, 6.0 g, 21.17 mmol) in 1,4-dioxane (200 mL) and water (20 mL). After stirring at 0 °C for 15 min, sodium periodate (18.1 g, 84.7 mmol) was added in portions to the reaction mixture at 0 °C, and the resulting mixture was warmed to 20 °C and stirred for 2 h. The reaction was quenched with saturated aqueous Na_₂SO_₃ solution (200 mL), and the reaction mixture was extracted three times with EtOAc (100 mL). The combined organic layers were washed with brine (100 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum to give _O₁ -tert- _butylO₄ -methyl 4-(2-oxoethyl)piperidine-1,4-dicarboxylate (compound 15C, 6.0 g), which was a yellow oil. _MS calculated value 286.2 ( ^MH⁺ ); measured value 186.2 ( _{MC₅H₈O₂} + _H⁺ ⁾ .

Step 3: Preparation of _O1 -tert- _butylO4 -methyl4-[2-[[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]amino]ethyl]piperidine-1,4-dicarboxylate (compound 15D)

Zinc chloride (1.4 g, 10.51 mmol) was added to a mixture of (2S)-2-amino-3-methyl-butyrate benzyl chloride (2.2 g, 10.51 mmol) and _O1 - _tert -butylO4-methyl4-(2-oxoethyl)piperidine-1,4-dicarboxylate (compound 15C, 2.0 g, 7.01 mmol) in methanol (30 mL) at 0 °C. After stirring at 0 °C for 0.5 h, sodium cyanoborohydride (0.9 g, 14.02 mmol) was added to the reaction mixture. After stirring at 20 °C for another 1 h, the reaction was quenched with saturated _NH4Cl aqueous solution (100 mL), and the reaction mixture was extracted three times with EtOAc (50 mL). The combined organic layers were washed with brine (50 mL), dried _over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum to obtain a residue, which was purified by column chromatography (EtOAc = 10% to 30% in PE) to give _O₁ -tert-butylO₄-methyl _4- [2-[[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]amino]ethyl]piperidine-1,4-dicarboxylate (compound 15D, 2.46 g), as a yellow oil. MS calculated value 477.3 (MH ⁺ ); measured value 477.3 (MH ⁺ ).

Step 4: Preparation of tert-butyl 2-[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylate (compound 15E)

DIEA (4.4 mL, 25.18 mmol) and DMAP (0.3 g, 2.52 mmol) were added to a mixture of 1-(tert-butyl)-4-methyl(S)-4-(2-((1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)amino)ethyl)piperidine-1,4-dicarboxylate (compound 15D, 1.2 g, 2.52 mmol) in toluene (20 mL). The reaction mixture was heated to 100 °C and stirred for 30 hours. After the reaction was complete, the reaction mixture was poured into water (30 mL) and extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine (30 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by reverse-phase column chromatography to give tert-butyl 2-[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylate (compound 15E, 586.0 mg), a yellow oil. MS calculated value 445.3 (MH ⁺ ); measured value 467.4 (MNa ⁺ ).

Step 5: Preparation of (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 15F)

A solution of tert-butyl 2-[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylic acid (compound 15E, 500.0 mg) in toluene (20 mL) was added to activated carbon-supported palladium (50.0 mg, 10% wt). The mixture was degassed and purged three times with _H₂ . The mixture was hydrogenated at 35 °C. The reaction solution was filtered and the filter cake was washed with DMF (100 mL). The combined filtrates were concentrated under vacuum to give (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 15F, 144.0 mg) as a white solid. MS calculated value 355.2 ( ^MH⁺ ); measured value 377.1 ( ^MNa⁺ ).

Examples 16 and 18

(2S)-2-[(5S)-2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butyramide (Example 16) and (2S)-2-[(5R)-2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5] .1 ^9,13 .0 ^22,26 ] Octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide (Example 18)

Similar to the preparation of Example 6, the title compound was prepared by using (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o 1) instead of (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J). Example 16 (14.0 mg) was obtained as a white solid. MS calculated value 1131.5 (MH ⁺ ); measured value 1131.4 (MH ⁺ ). ¹ H NMR (400MHz, methanol-d4) δ＝8.69(dd,J＝2.0,7.6Hz,1H),8.50(d,J＝2.8Hz,1H),7.80-7.70(m,1H),7.51-7.45(m,2H),6.91-6.73(m,1H),5.72-5.64(m,1H),5.23-5.13(m,2H),4.42(d,J＝12.0Hz,1H),4.27-4.20(m,2H),4.10-4.00(m,3H),3.93-3.82(m,2H),3.79-3.69(m,3H),3.66-3.56 (m,3H),3.52-3.43(m,4H),3.36-3.34(m,4H),3.19-3.12(m,2H),3.0 0(s,3H),2.87-2.79(m,1H),2.60-2.53(m,1H),2.52-2.41(m,1H),2.3 9-2.18(m,3H),2.07-1.81(m,8H),1.68-1.61(m,1H),1.45(d,J＝6.0Hz,3H),1.00-0.96(m,6H),0.90(d,J＝6.4Hz,3H),0.47-0.41(m,3H)ppm.

Similar to the preparation of Example 6, the title compound was prepared by using (2S)-2-[(5R)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o 2) instead of (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J). Example 18 (25.6 mg) was obtained as a white solid. MS calculated value 1131.4 (MH ⁺ ); measured value 1131.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.68(d,J＝7.6Hz,1H),8.50(d,J＝2.8Hz,1H),7.74-7.71(m,1H),7.50-7.45(m,2H),6.91-6.76(m,1H),5.72-5.65 (m,1H),5.24-5.11(m,2H),4.83-4.79(m,3H),4.47-4.18(m,2H),4.12-3.87(m,3H),3.81-3.60(m,6H),3.56-3.44(m ,3H),3.41-3.33(m,6H),3.26-3.12(m,3H),3.00(s,3H),2.87-2.78(m,1H),2.26-2.46(m,2H),2.39-2.19(m,2H),2. 05-1.79(m,7H),1.71-1.60(m,1H),1.44(d,J＝6.4Hz,3H),1.02-0.95(m,6H),0.90(d,J＝6.4Hz,3H),0.44(s,3H)ppm.

Compounds 16o 1 and 16o 2 were 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 16c)

Under a nitrogen atmosphere, at -70 °C, LDA (59.9 mL, 119.95 mmol) was added to a solution of 1-(tert-butyl)-3-methylpyrrolidine-1,3-dicarboxylate (compound 16a, 25.0 g, 109.04 mmol) in 300 mL of THF. After stirring for 0.5 h, 4-bromo-1-butene (compound 16b, 16.1 g, 119.95 mmol) was added to the reaction mixture. After stirring at 20 °C for 2.5 h, the reaction was quenched with saturated _NH4Cl (100 mL), and the reaction mixture was extracted twice with EtOAc (150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which was purified by silica gel column chromatography (EtOAc = 0%–5% in PE) to give 1-(tert-butyl)3-methyl-3-(but-3-en-1-yl)pyrrolidine-1,3-dicarboxylate (compound 16c, 13.2 g), which is a yellow oil. ^¹H 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 16d)

A solution of HCl (50.0 mL, 4 M, in dioxane) was added to a solution of 1-(tert-butyl)-3-methyl-3-(but-3-en-1-yl)pyrrolidine-1,3-dicarboxylate (compound 16c, 13.2 g, 46.58 mmol) in 1,4-dioxane (50 mL). After stirring at 20 °C for 0.5 h, the reaction mixture was concentrated under vacuum to give methyl 3-(but-3-en-1-yl)pyrrolidine-3-carboxylate hydrochloride (compound 16d, 10.2 g), which is a yellow oil.

Step 3: Preparation of methyl 3-(but-3-en-1-yl)-1-triphenylmethylpyrrolidone-3-carboxylate (compound 16e)

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 16d, 3.0 g, 13.65 mmol) in ACN (70 mL). The mixture was stirred at 20 °C for 12 hours. After the reaction was complete, the mixture was poured into water (100 mL) and extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a residue, which was purified by silica gel column chromatography (EtOAc = 0%–5% in PE) to give methyl 3-(but-3-en-1-yl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 16e, 1.3 g), which is a yellow oil. ¹ H NMR (400MHz, CDCl ₃ )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,1 H),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 16f)

Potassium osmium tetroxide (VI) (5 mg, 0.02 mmol) and sodium periodate (663 mg, 3.1 mmol) were added to a solution of methyl 3-(but-3-en-1-yl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 16e, 660 mg, 1.55 mmol) in THF (20 mL) and water (10 mL). The reaction mixture was stirred at 20 °C for 2 hours. After the reaction was complete, it was quenched with a saturated aqueous solution of _NH4Cl , and the reaction mixture was extracted twice with EtOAc (50 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a residue, which was purified by silica gel column chromatography (EtOAc in PE = 0%–5%) to give methyl 3-(3-oxopropyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 16f, 300 mg), which is 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 16h)

Sodium cyanoborohydride (88.9 mg, 1.42 mmol) was added to a mixture of methyl 3-(3-oxopropyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 16f, 550.0 mg, 1.29 mmol), H-VAL-OTBU HCl (compound 16 g, 296.7 mg, 1.42 mmol), and zinc chloride (192.8 mg, 1.42 mmol) in methanol (6 mL) at 0 °C. The mixture was stirred at 25 °C for 2 hours. After the reaction was complete, water (30 mL) was added to the reaction mixture. The resulting mixture was concentrated under vacuum. The resulting suspension was diluted with water (40 mL) and extracted twice with EtOAc (30 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by column chromatography (EtOAc = 0%–10% in PE) to give methyl 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutane-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylate (compound 16h, 510 mg), which is a yellow oil. ^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 16i)

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 16h, 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 combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutane-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylic acid (compound 16i, 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 16j)

At 0 °C, DIEA (0.67 mL, 3.85 mmol) and COMU (594 mg, 1.39 mmol) were added to a solution of 3-(3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl)amino)propyl)-1-triphenylmethylpyrrolidine-3-carboxylic acid (compound 16i, 440 mg, 0.77 mmol) in DMF (5 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was poured into water (20 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which was purified by silica gel column chromatography (EtOAc = 0%–10% in PE) to give tert-butyl (2S)-3-methyl-2-(6-oxo-2-triphenylmethyl-2,7-diazaspiro[4.5]decane-7-yl)butyrate (compound 16j, 420 mg), as 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 16k)

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 16j, 550 mg, 1 mmol) in DCM (3 mL). The mixture was stirred at 20 °C for 12 hours. After the reaction was complete, the mixture was concentrated under vacuum to obtain a residue, which was dissolved in water (10 mL), and the aqueous phase was washed 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)butyric acid (compound 16k, 365 mg) as a TFA salt, which was a colorless oil. 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 16l).

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 16k, 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 the reaction mixture was extracted twice with EtOAc (20 mL). The combined organic layers were 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 16k, 350 mg) as a white solid.

Step 10: Preparation of (5S)-7-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (compound 16m 1) and (5R)-7-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (compound 16m 2)

7-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (compound 16l, 680.0 mg, 1.66 mmol) was passed through an SFC (regis(S,S)whelk-O1 (250 mm, twice, 5 mm × 10 μm) mobile phase: (0.1% _NH3 H2 ₎ (O)IPA, B%: 15%-15%, 4.4 min) separation to obtain (5S)-7-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (compound 16 ml, 245.0 mg, fast elution), which is a yellow oil; and (5R)-7-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (compound 16 ml, 223.0 mg, slow elution), which is a yellow oil. MS calculated value 411.3 (MH ⁺ ); measured value 411.3 (MH ⁺ ).

Step 11: Preparation of (2S)-3-methyl-2-[(5S)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]butyric acid (compound 16n 1)

TFA (1.0 mL, 12.98 mmol) was added to a solution of (5S)-7-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (compound 16m 1, 245.0 mg, 0.6 mmol) in DCM (1 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the mixture was concentrated under vacuum to give (2S)-3-methyl-2-[(5S)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]butyric acid (compound 16n 1, 219.0 mg, TFA salt), a colorless liquid, which was used directly in the next step. MS calculated value 255.2 (MH ⁺ ); measured value 255.2 (MH ⁺ ).

Step 12: Preparation of (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o1)

Sodium carbonate (189.05 mg, 1.78 mmol) and di-tert-butyl dicarbonate (155.71 mg, 0.71 mmol) were added to a solution of (2S)-3-methyl-2-[(5S)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]butyric acid (compound 16n 1, 219.0 mg, 0.59 mmol) in THF (2 mL) and water (2 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the mixture was filtered and the filtrate was concentrated under vacuum to obtain a residue, which was purified by reverse-phase rapid column chromatography to give (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o 1, 200.0 mg), which was a white solid. MS calculated value 353.2 (MH ^- ); measured value 353.0 (MH ^- ).

Step 13: Preparation of (2S)-3-methyl-2-[(5R)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]butyric acid (compound 16n 2)

TFA (1.5 mL, 19.47 mmol) was added to a solution of (5R)-7-[(1S)-1-tert-butoxycarbonyl-2-methyl-propyl]-6-oxo-2,7-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (compound 16m 2, 223.0 mg, 0.54 mmol) in DCM (1.5 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the mixture was concentrated under vacuum to give (2S)-3-methyl-2-[(5R)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]butyric acid (compound 16n 2, 200.0 mg, TFA salt), a colorless liquid, which was used directly in the next step. MS calculated value 255.2 (MH ⁺ ); measured value 255.1 (MH ⁺ ).

Step 14: Preparation of (2S)-2-[(5R)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o 2)

Sodium carbonate (115.1 mg, 1.09 mmol) and di-tert-butyl dicarbonate (142.2 mg, 0.65 mmol) were added to a solution of (2S)-3-methyl-2-[(5R)-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]butyric acid (compound 16n 2, 200.0 mg, 0.54 mmol) in THF (2 mL) and water (2 mL). The mixture was stirred at 20 °C for 1 h. After the reaction was complete, the mixture was filtered and the filtrate was concentrated under vacuum to obtain a residue, which was purified by reverse-phase rapid column chromatography to give (2S)-2-[(5R)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o 2, 150.0 mg), which was a white solid. MS calculated value 355.2 (MH ⁺ ); measured value 377.2 (MNa ⁺ ).

Example 20

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 6, this was achieved by using (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 20 (6.1 mg) was obtained as a white solid. MS calculated value 1145.5 (MH ⁺ ); measured value 1145.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4 ) δ＝8.52(s, 1H), 7.61–7.45(m, 4H), 7.09–6.91(m, 1H), 5.97–5.84(m, 1H), 5.24–5.06(m, 3H), 4.69–4.56(m, 0.5H), 4.44–4.33(m, 0.5H), 4.20–3.96(m, 3H), 3.90–3.40(m, 10H), 3.27–3.10(m, 12H), 3.00(s, 3H). H),2.95-2.75(m,1H),2.71-2.53(m,1H),2.25-2.04(m,2H),2.00-1.79(m,4H),1.72-1.53(m,3H),1.4 4(d,J＝6.0Hz,3H),1.37-1.19(m,2H),0.94(d,J＝6.0Hz,3H),0.89-0.73(m,7H),0.71-0.53(m,3H)ppm.

Examples 22 and 23

(2S)-2-[(5R)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butyramide (Example 22) and (2S)-2-[(5S)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0] ^22,26 ] Octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide (Example 23)

Similar to the preparation in Example 6, this was achieved by using (2S)-2-chloro-2-fluoroacetic acid, (2S)-2-[(5S)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o 1), and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate H) replaces (2R)-2-chloro-2-fluoroacetic acid, (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1] The title compound was prepared from ^octadecano -1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 22 ( ^4.1 mg) was obtained as a white solid. MS calculated value 1077 (MH ⁺ ); measured value 1077 (MH ⁺ ). ¹ HNMR (400MHz, DMSO-d ₆ ) δ 9.98-10.20 (m, 1H), 8.42-8.62 (m, 2H), 8.10-8.34 (m, 1H), 7.71 (d, J = 2.50Hz, 1H), 7.57 (d, J = 13.01Hz, 1H), 7.39 (br s,1H),7.15(d,J＝10.01Hz,1H),7.00-7.07(m,1H),5.41-5.54(m,1H),5.18(br dd,J＝19.51,11.88Hz,1H),4.81(br dd,J＝12.88,11.38Hz,1H),4.23-4.28(m,2H),4.13(br s,5H),3.57(br s,3H),3.34-3.34(m,3H),2.82-2.94(m,3H),2.66-2.81(m,7H),2.41(br s,1H),2.31-2.34(m,3H),2.21(br d,J＝6.88Hz,1H),1.69-1.88(m,7H),1.50(br s,1H),1.33(d,J＝6.13Hz,3H),1.18-1.26(m,3H),0.84-0.98(m,11H),0.80(br t,J＝7.32Hz,3H),0.35(s,3H)ppm.

Similar to the preparation in Example 6, this was achieved by using (2S)-2-chloro-2-fluoroacetic acid, (2S)-2-[(5R)-2-tert-butoxycarbonyl-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-3-methyl-butyric acid (compound 16o 2), and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate H) replaces (2R)-2-chloro-2-fluoroacetic acid, (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1] Example ²³ was prepared using octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D) ^. Example 23 (2.2 mg) was obtained as a white solid. MS calculated value 1077 (MH ⁺ ); measured value 1077 (MH ⁺ ). ¹ H NMR (400MHz, methanol-d ₄ ) δppm 8.66 (d, J = 7.50Hz, 1H), 8.47 (d, J = 2.88Hz, 1H), 7.66 (br d,J＝7.13Hz,1H),7.45(d,J＝2.75Hz,1H),7.33-7.36(m,1H),6.73-6.94(m,1H),5. 32-5.38(m,2H),4.37-4.53(m,2H),4.09-4.30(m,5H),3.68-3.81(m,4H),3.61(br d,J＝7.00Hz,4H),3.04(q,J＝7.75Hz,4H),2.94(br s,3H),2.77-2.87(m,2H),2.54-2.66(m,2H),2.16-2.22(m,3H),2.01-2.05(m,3H),1.88(br s,2H),1.63(dt,J＝15.26,4.31Hz,8H),1.43(d,J＝6.25Hz,3H),0.89-1.00(m,15H),0.49(br d,J＝2.63Hz,3H).

Example 24

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

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate E) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E). Example 24 (18.9 mg) was obtained as a white solid. MS calculated value 1092.4 (MH ⁺ ); measured value 1092.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ = 8.79–8.66 (m, 1H), 8.60–8.35 (m, 1H), 7.81–7.67 (m, 2H), 7.57–7.42 (m, 1H), 6.94–6.72 (m, 1H), 5.76–5.61 (m, 1H), 5.33–5.10 (m, 1H), 4.80–4.60 (m, 2H), 4.43–4.10 (m, 3H), 3.88–3.5 2(m,12H),3.43-3.36(m,8H),3.11-2.99(m,4H),2.85-2.81(m,1H),2.68-2.64(m,1H),2.34- 2.21(m,4H),1.95-1.61(m,3H),1.48-1.47(m,3H),1.07-0.81(m,9H),0.60-0.42(m,3H)ppm.

Example 25

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

Similar to the preparation of Example 6, the title compound was prepared by using (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 25H) instead of (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J). Example 25 (20.8 mg) was obtained as a white solid. MS calculated value 1103.4 (MH ⁺ ); measured value 1103.7 (MH ⁺ ). ^¹H NMR (400MHz, methanol- _d⁴ ) δ＝8.96(d,J＝8.4Hz,1H),8.68(d,J＝7.6Hz,1H),8.50(d,J＝2.8Hz,1H),7.72(dd,J＝2.4,17.6Hz,1H),7.54-7.41(m,2H),6.80-6.61(m,1H),5.72-5.60(m,1H),5.21

-5.14(m,1H),4.65-4.55(m,1H),4.45-4.38(m,2H),4.38-4.28(m,2H),4.26

-4.19(m,2H),4.16-4.00(m,3H),3.81-3.67(m,3H),3.66-3.53(m,3H),3.51

-3.39(m,4H),3.35(s,3H),3.25-3.10(m,3H),3.00(s,3H),2.89-2.78(m ,1H),2.60-2.52(m,1H),2.47-2.36(m,2H),2.28-2.19(m,2H),1.99-1.92 (m,1H),1.89-1.78(m,1H),1.70-1.59(m,1H),1.44(d,J＝6.0Hz,3H),1.0 1(dd,J=3.6,6.4Hz,3H),0.97(s,3H),0.92-0.86(m,3H),0.43(s,3H)ppm.

Compound 25H was prepared according to the following scheme:

Step 1: tert-butyl 3-allyl-3-cyano-azacyclobutane-1-carboxylate (compound 25B)

Under a nitrogen atmosphere, at -70 °C, LDA (60.3 mL, 60.37 mmol) was added dropwise to a solution of tert-butyl 3-cyano-azacyclobutane-1-carboxylate (compound 25A, 10.0 g, 54.88 mmol) in 100 mL of THF. After stirring at -70 °C for 0.5 h, allyl bromide (7.9 g, 65.85 mmol) was added to the reaction mixture. The mixture was stirred at 16 °C for another 1 h, then concentrated under vacuum, and the residue was purified by column chromatography (EtOAc in PE: 3%–10%) to give tert-butyl 3-allyl-3-cyano-azacyclobutane-1-carboxylate (compound 25B, 12.5 g), which is a pale yellow oil. ¹ H NMR (400 MHz, chloroform-d) δ 5.90–5.70 (m, 1H), 5.32 (s, 1H), 5.28 (d, J = 5.2 Hz, 1H), 4.23 (d, J = 8.8 Hz, 2H), 3.86 (d, J = 8.8 Hz, 2H), 2.63 (d, J = 7.2 Hz, 2H), 1.45 (s, 9H) ppm.

Step 2: 3-Allyl-1-tert-butoxycarbonyl-azacyclobutane-3-carboxylic acid (compound 25C)

A mixture of tert-butyl 3-allyl-3-cyano-azacyclobutane-1-carboxylic acid (compound 25B, 12.5 g, 56.24 mmol) and potassium hydroxide (12.6 g, 224.94 mmol) in ethanol (60 mL) and water (60 mL) was stirred at 100 °C for 16 h. After the reaction was complete, the pH of the reaction mixture was acidified to pH 3 with an aqueous solution of HCl (1 M). The reaction mixture was extracted three times with EtOAc (120 mL). The combined organic layers were washed with brine ( ₃₀₀ mL), dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give 3-allyl-1-tert-butoxycarbonyl-azacyclobutane-3-carboxylic acid (compound 25C, 14.0 ^g ) as a white solid. MS calculated value 241.1 ( ^MH⁺ ); measured value _186.2 ( _{MC₄H₈⁺H⁺} ).

Step 3: _O1 -tert-butylO3- _methyl3 -allylazyrobutane-1,3-dicarboxylate (compound 25D)

Potassium carbonate (13.7 g, 99.47 mmol) was added to a mixture of 3-allyl-1-tert-butoxycarbonyl-aziridine-3-carboxylic acid (compound 25C, 8.0 g, 33.16 mmol) in DMF (80 mL), followed by iodomethane (9.4 g, 66.31 mmol). After stirring at 20 °C for 2 hours, the reaction mixture was poured into water (400 mL), and the resulting mixture was extracted three times with EtOAc (100 mL). The combined organic layers were washed with brine (300 mL ₎ , dried over _Na₂SO₄ , filtered, and concentrated under vacuum to give _O₁ -tert-butylO₃- _methyl3 -allylaziridine-1,3-dicarboxylic acid ester (compound 25D, 7.3 g), which is a yellow oil. ^1H NMR (400MHz, chloroform-d) δ 5.80-5.60 (m, 1H), 5.19-5.14 (m, 1H), 5.12 (s, 1H), 4.16 (d, J = 8.8Hz, 2H), 3.82-3.69 (m, 5H), 2.63 (d, J = 7.2Hz, 2H), 1.44 (s, 9H) ppm.

Step 4: _O1 -tert- _butylO3 -methyl 3-(2-oxoethyl)azacyclobutane-1,3-dicarboxylate (compound 25E)

2,6-Dimethylpyridine (1.8 mL, 15.67 mmol) and K₂O₅sO₄ (144.3 mg, 0.39 mmol) were added to a mixture of _O₁ -tert- _butylO₃ -methyl- ₃ -allylazyrazine-1,3-dicarboxylate (compound 25D, 2 g, 7.83 mmol) in 1,4-dioxane (20 mL) and water ( ₂₀ mL). After stirring at 16 °C for 15 min, sodium periodate (6702.2 mg, 31.33 mmol) was added to the reaction mixture and stirred for another 1 h. The mixture was then poured into water (120 mL), and the resulting mixture was extracted three times with EtOAc (40 mL). The combined organic layers were washed with saturated _Na₂SO₃ (60 mL) _and brine (60 mL), dried _{over Na₂SO₄} , filtered, and concentrated under vacuum to give _O₁ -tert- _butylO₃ -methyl 3-(2-oxoethyl)azacyclobutane-1,3-dicarboxylate (compound 25E, 2.8 g), which was a yellow oil. MS calculated value 257.1 ( ^MH⁺ ); measured value _202.1 ( _{MC₄H₈⁺H⁺} ⁾ .

Step 5: _O1 -tert- _butylO3 -methyl3-[2-[[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]amino]ethyl]azacyclobutane-1,3-dicarboxylate (compound 25F)

Zinc chloride (1.8 g, 13.06 mmol) was added to a mixture of L-valine benzyl ester hydrochloride (3.2 g, 13.06 mmol) and _O1 - _tert -butylO3-methyl 3-(2-oxoethyl)azacyclobutane-1,3-dicarboxylate (compound 25E, 2.8 g, 10.88 mmol) in methanol (50 mL). After stirring at 16 °C for 0.5 h, sodium cyanoborohydride (1.4 g, 21.77 mmol) was added to the reaction mixture, and the resulting mixture was stirred for another 1 h. The reaction mixture was concentrated under vacuum, and the residue was purified by column chromatography (EtOAc in PE: 30%–50%) to give _O1 -tert-butylO3-methyl _3- [2-[[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]amino]ethyl]azacyclobutane-1,3-dicarboxylate (compound 25F, 2.2 g), as a colorless oil. MS calculated value 449.3 (MH ⁺ ); measured value 449.3 (MH ⁺ ).

Step 6: 6-[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]-5-oxo-2,6-diazaspiro[3,4]octane-2-carboxylic acid tert-butyl ester (compound 25G)

A mixture of 4-dimethylaminopyridine (544.7 mg, 4.46 mmol), DIEA (7.7 mL, 44.59 mmol), and _O1 -tert-butylO3-methyl _3- [2-[[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]amino]ethyl]azacyclobutane-1,3-dicarboxylate (compound 25F, 2.0 g, 4.46 mmol) in toluene (2 mL) was stirred at 100 °C for 20 hours. After the reaction was complete, the reaction mixture was concentrated under vacuum, and the residue was dissolved in EtOAc (60 mL). The resulting solution was washed with aqueous HCl solution (1 M, 60 mL), then washed with brine (60 _mL ), dried over _Na2SO4 , and filtered. The filtrate was concentrated under vacuum to give tert-butyl 6-[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]-5-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (compound 25g, 1.5g), a white solid. MS calculated value 417.2 (MH ⁺ ); measured value 361.2 ( _MC4H8 + _H ⁺ ).

Step 7: (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octane-6-yl)-3-methyl-butyric acid (compound 25H)

Under _H₂ balloon conditions, a mixture of tert-butyl 6-[(1S)-1-benzyloxycarbonyl-2-methyl-propyl]-5-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (compound 25 G, 1.5 g, 3.6 mmol) and activated carbon-supported Pd (150.0 mg) in methanol (20 mL) was stirred at 16 °C for 2 h. The reaction mixture was filtered, and the filtrate was concentrated under vacuum to give (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octane-6-yl)-3-methyl-butyric acid (compound 25 H, 920.0 mg), a white solid. MS calculated value 327.2 ( ^MH⁺ ); measured value _271.2 ( _{MC₄H₈⁺H⁺} ⁾ .

Example 26

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

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22 ,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E) with (7S,13S)-7-amino-25-fluoro-(20M)-20-[ ² -[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 ^9,13 .0 ^{22 ,26} ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E). Example 26 (14.6 mg) was obtained as a white solid. MS calculated value 1173.4 (MH ⁺ ); measured value 1173.7 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ＝8.72-8.68(m,1H),8.42(d,J＝2.4Hz,1H),7.77-7.68m,1H),7.57-7.45(m,2H),6.93-6.76(m,1H),5.81-5.63(m,1H),5 .27-5.11(m,1H),4.96-4.93(m,1H),4.88-4.79(m,2H),4.47-4.38(m,1H),4.28-4.18(m,2H),3.88-3.53(m,7H),3.47-3. 34(m,8H),3.30-3.24(m,1H),3.23-3.07(m,6H),2.92-2.80(m,5H),2.70-2.56(m,1H),2.40-2.06(m,4H),2.00-1.91(m,1 H),1.88-1.74(m,1H),1.69-1.57(m,1H),1.49-1.41(m,3H),1.01-0.97(m,5H),0.92-0.82(m,3H),0.53-0.42(m,3H)ppm.

Example 27

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-26-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^{23 ,27]} [26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

Similar to the preparation in Example 1, this was achieved by using (8S,14S)-8-amino-26-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadecane-1(25), ^2,5 (28), ^19,22 (26),23-hexaene-8,14-dione (intermediate E) with 29-carbon-1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (intermediate K). Example 27 (5.2 mg) was obtained as a white solid. MS calculated value 1114.5 (MH ⁺ ); measured value 1114.6 (MH ⁺ ), ^1H NMR (400 MHz, methanol- _d4) )δ:8.50(d,J＝2.8Hz,1H),8.21-8.10(m,1H),7.51(s,1H),7.47(d,J＝8.0Hz,1H),7.42-7.36(m,1H),7.14-7.06(m,1H),6.89-6.86(m,1H),5 .34(t,J＝4.4Hz,1H),4.69(d,J＝10.8Hz,1H),4.43-4.34(m,0.5H),4.27-4.17(m,0.5H),4.14-3.95(m,1H),3.79-3.56(m,6H),3.23(s,2H), 3.02(d,J＝14Hz,6H),2.91-2.78(m,2H),2.78-2.56(m,2H),2.19(t,J＝7.6Hz,2H),2.06-2.01(m,3H),1.88-1.77(m,1H),1.64-1.56(m,1H), 1.50-1.46(m,3H),1.44(d,J＝1.6Hz,1H),1.34-1.29(m,10H),1.18(t,J＝6.8Hz,2H),0.97-0.87(m,8H),0.86-0.82(m,3H),0.41(s,3H)ppm.

Example 28

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 15F) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate H) replaces (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 28 (11.7 mg) was obtained as a white solid. MS calculated value 1077.5 (MH ⁺ ); measured value 1077.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ8.64(d,J＝7.7Hz,1H),8.43(d,J＝1.7Hz,1H),7.66(s,1H),7.43(d,J＝2.2Hz,1H),7.30(d,J＝12.3Hz,1H),7 .13-6.92(m,1H),5.69-5.53(m,2H),5.51-5.36(m,1H),4.58(s,11H),4.54-4.23(m,1H),4.22-3.80(m,1H), 3.78-3.46(m,3H),2.75-2.59(m,5H),2.44-2.30(m,5H),2.29-2.19(m,2H),2.18-2.05(m,3H),2.03-1.74(m ,6H),1.70-1.55(m,3H),1.30-1.15(m,7H),1.03-0.94(m,7H),0.90(dd,J=2.6,6.5Hz,3H),0.61(s,3H)ppm.

Example 29

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octane-6-yl)-3-methyl-butyric acid (compound 25H) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate H) replaces (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 29 (63.2 mg) was obtained as a yellow solid. MS calculated value 1049.5 (MH ⁺ ); measured value 1049.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ=8.76-8.63(m,1H),8.58-8.45(m,1H),7.78-7.56(m,2H),7.44-7.30(m,1H),6.82-6.59(m,1H),5.83-5.67(m,1H),4.65-4.53( m,1H),4.50-3.99(m,11H),3.88-3.67(m,3H),3.61-3.47(m,3H),3.47-3.42(m,3H),3.41-3.35(m,4H),3.32-3.22(m,2H),3.11-3 .04(m,1H),3.01(s,3H),2.90-2.78(m,1H),2.69-2.60(m,1H),2.45-2.35(m,2H),2.31-2.16(m,2H),2.03-1.92(m,1H),1.90-1.7 5(m,1H),1.72-1.58(m,1H),1.52-1.41(m,3H),1.08-0.98(m,6H),0.96(s,3H),0.89(dd,J＝2.8,6.4Hz,3H),0.62-0.43(m,3H)ppm.

Examples 30 and 31

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 30)

(Example 30)

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 31)

(Example 31)

Similar to the preparation in Example 4, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Example 30 was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 30 (42.4 mg) was obtained as a yellow solid. MS calculated value 1063.5 (MH ⁺ ); measured value 1063.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ8.72-8.65(m,1H),8.56-8.45(m,1H),7.78-7.73(m,0.5H),7.70-7.64(m,0.5H),7.63-7.57(m,1H),7.41-7.31(m,1H),6.95-6.71(m,1H ),5.82-5.67(m,1H),4.51-4.02(m,8H),3.99-3.90(m,1H),3.88-3.70(m,5H),3.69-3.56(m,4H),3.52-3.43(m,3H),3.37(d,J＝6.4Hz,4H), 3.30-3.23(m,2H),3.13-3.05(m,1H),3.01(s,3H),2.88-2.79(m,1H) ,2.69-2.57(m,1H),2.36-2.18(m,3H),2.18-2.04(m,3H),2.01-1.96 (m,1H),1.88-1.76(m,1H),1.71-1.59(m,1H),1.46(d,J＝6.0Hz,3H),1.05-0.94(m,9H),0.94-0.89(d,J＝6.6Hz,3H),0.58-0.41(m,3H)ppm.

Similar to the preparation in Example 4, this was achieved by using (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate H) replaces (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13]} Example ³¹ was prepared using octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 31 (56.7 mg) was obtained as a yellow solid. MS calculated value 1063.5 (MH ⁺ ); measured value 1063.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.67(d,J＝7.6Hz,1H),8.48(d,J＝2.8Hz,1H),7.65(s,1H),7.57(s,1H),7.34(d,J＝12.8Hz,1H),6.94-6.73(m,1H),5.75(t,J＝8.8Hz,1H),4. 45-4.37(m,1H),4.36-4.29(m,2H),4.28-4.08(m,4H),4.07-3.97(m,1H ),3.97-3.87(m,1H),3.85-3.72(m,4H),3.71-3.63(m,2H),3.63-3.41( m,7H),3.34(s,3H),3.30-3.23(m,2H),3.07-3.02(m,1H),3.00(s,3H), 2.87-2.77(m,1H),2.69-2.57(m,1H),2.33-2.15(m,3H),2.14-2.03(m, 3H),1.97-1.91(m,1H),1.84-1.72(m,1H),1.69-1.57(m,1H),1.44(d,J ＝6.0Hz,3H),1.03-0.92(m,9H),0.89(d,J＝6.8Hz,3H),0.51(s,3H)ppm.

Example 32

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E) with (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E). Example 32 (36.7 mg) was obtained as a yellow solid. MS calculated value 1051.4 (MH ⁺ ); measured value 1051.6 (MH ⁺ ). ¹ H NMR (400MHz, MeOD) δ8.67(d,J＝7.6Hz,1H),8.49(d,J＝2.8Hz,1H),7.73-7.59(m,2H),7.34(d,J＝12.8Hz,1H),6.96-6.70(m,1H),5.82-5.61(m,1H),5 .00-4.90(m,2H),4.83-4.77(m,1H),4.49-4.38(m,1H),4.37-4.05(m,5H) ,4.00-3.89(m,1H),3.89-3.47(m,10H),3.47-3.39(m,2H),3.36(d,J＝4.8 Hz,3H),3.28-3.22(m,1H),3.15-3.07(m,3H),3.06-3.02(m,1H),3.00(s, 3H),2.89-2.76(m,1H),2.69-2.56(m,1H),2.42-2.01(m,4H),2.00-1.90( m,1H),1.89-1.73(m,1H),1.71-1.56(m,1H),1.44(d,J＝6.4Hz,3H),1.25- 0.96(m,6H),0.96-0.92(m,3H),0.92-0.82(m,3H),0.61-0.39(m,3H)ppm.

Example 33

(3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26 ] [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide]

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate E) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E). Example 33 (45.2 mg) was obtained as a yellow solid. MS calculated value 1119.5 (MH ⁺ ); measured value 1119.7 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ＝8.73-8.65(m,1H),8.38(s,1H),7.94-7.86(m,1H),7.72-7.62(m,1H),7.37(d,J＝12.6Hz,1H),6.94-6.72(m,1H),5.83-5.6 9(m,1H),4.84-4.76(m,1H),4.47-4.19(m,3H),4.17-3.92(m,2H),3.89-3.62(m,6H),3.61-3.53(m,1H),3.51-3.46(m,4H),3. 43-3.37(m,3H),3.28-3.13(m,3H),3.10-3.03(m,3H),2.93-2.87(m,4H),2.86-2.66(m,2H),2.42-2.11(m,4H),2.00-1.90(m, 1H),1.86-1.73(m,1H),1.71-1.57(m,1H),1.46(d,J=6.4Hz,3H),1.36-1.27(m,1H),1.14-0.78(m,13H),0.69-0.54(m,3H)ppm.

Example 34

1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidin-4-carboxamide

Similar to the preparation in Example 1, 1-tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate H) replaces (S)-1-Boc-pyrrolidine-3-carboxylic acid (compound 1D) and (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E). Example 34 (12.9 mg) was obtained as a yellow solid. MS calculated value 1083.5 (MH ⁺ ); measured value 1083.6 (MH ⁺ ), ^1H NMR (400 MHz, methanol- _d4) )δ＝8.65(d,J＝7.6Hz,1H),8.48(d,J＝2.4Hz,1H),7.64(s,1H),7.54(s,1 H),7.34(d,J＝12.4Hz,1H),7.11-6.95(m,1H),5.78-5.71(m,1H),4.47- 4.37(m,2H),4.35-4.23(m,2H),4.22-4.07(m,4H),4.00-3.88(m,2H),3 .80-3.69(m,3H),3.68-3.57(m,2H),3.54-3.41(m,4H),3.34(s,3H),3.2 0(t,J＝4.8Hz,4H),3.14-3.05(m,2H),3.03-2.95(m,5H),2.86-2.79(m, 1H),2.65-2.57(m,1H),2.32-2.13(m,5H),2.03-1.90(m,2H),1.84-1.7 7(m,1H),1.68-1.59(m,1H),1.44(d,J＝6.0Hz,3H),1.29-1.09(m,2H),1 .00-0.97(m,3H),0.93(s,3H),0.91-0.86(m,3H),0.50(d,J＝5.2Hz,3H).

Examples 35 and 38

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 35)

(Example 35)

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 38)

(Example 38)

Similar to the preparation in Example 4, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Example 35 was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 35 (21.1 mg) was obtained as a yellow solid. MS calculated value 1131.5 (MH ⁺ ); measured value 1131.6 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ8.71(dd,J＝2.4,7.6Hz,1H),8.38(t,J＝2.4Hz,1H),7.87-7.81(m,1H),7.76-7.62(m,1H),7.37(dd,J＝2.8,12.6Hz,1H),6.92-6 .69(m,1H),5.84-5.71(m,1H),4.45-4.12(m,5H),4.10-4.00(m,1H),3.96-3.69(m,5H),3.60-3.54(m,2H),3.51-3.42(m,6H),3. 41-3.37(m,3H),3.26-3.12(m,3H),3.09-2.96(m,1H),2.93-2.87(m,4H),2.85-2.66(m,2H),2.34-2.15(m,3H),2.12-1.93(m,4H ),1.86-1.73(m,1H),1.71-1.57(m,1H),1.45(d,J＝6.4Hz,3H),1.04-0.95(m,9H),0.89(d,J＝6.8Hz,3H),0.58(d,J＝12.8Hz,3H).

Similar to the preparation in Example 4, this was achieved by using (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate L) replacing (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 Example ³⁸ was prepared using octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 38 (26.1 mg) was obtained as a yellow solid. MS calculated value 1131.5 (MH ⁺ ); measured value 1131.7 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.88-8.81(m,1H),8.71(d,J＝7.6Hz,1H),8.38(d,J＝2.0Hz,1H),7.9 6-7.87(m,1H),7.67-7.64(m,1H),7.37(d,J＝12.4Hz,1H),6.91-6.75(m, 1H),5.84-5.79(m,1H),4.46-4.36(m,2H),4.33-4.25(m,2H),4.18-4.12 (m,1H),4.08-3.84(m,2H),3.81-3.43(m,4H),3.69-3.61(m,2H),3.52-3 .48(m,5H),3.40(s,3H),3.23-3.15(m,3H),3.03-2.97(m,1H),2.91-2.8 8(m,4H),2.83-2.73(m,2H),2.27-2.15(m,3H),2.11-2.04(m,3H),2.00- 1.91(m,2H),1.85-1.76(m,1H),1.70-1.61(m,1H),1.46(d,J＝6.4Hz,3H) ,1.04-0.99(m,5H),0.96(s,3H),0.89(d,J＝6.8Hz,3H),0.61(s,3H)ppm.

Example 36

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadecane-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

Similar to the preparation in Example 6, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 36 (27.5 mg) was obtained as a yellow solid. MS calculated value 1091.5 (MH ⁺ ); measured value 1091.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ＝8.67(d,J＝7.6Hz,1H),8.48(d,J＝2.8Hz,1H),7.67(d,J＝2.4Hz,1H),7.61-7.51(m,1H),7.34(d,J＝12.8Hz,1H),7.15-6.85(m,1H),5. 80-5.68(m,1H),4.86-4.76(m,1H),4.47-4.36(m,1H),4.35-4.05(m,6H),3.97-3.86(m,1H),3.83-3.67(m,3H),3.66-3.51(m,2H),3.50- 3.40(m,4H),3.34(s,3H),3.30-3.09(m,4H),3.07-3.01(m,1H),3.00(s,3H),2.88-2.76(m,1H),2.68-2.56(m,1H),2.33-2.08(m,4H),1 .97-1.75(m,6H),1.68-1.57(m,3H),1.44(d,J＝6.0Hz,3H),1.37-1.24(m,2H),1.01-0.90(m,9H),0.87(d,J＝6.4Hz,3H),0.51(s,3H)ppm.

Example 37

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octane-6-yl)-3-methyl-butyric acid (compound 25H) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate L) replaces (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 37 (32.1 mg) was obtained as a yellow solid. MS calculated value 1117.5 (MH ⁺ ); measured value 1117.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.69(d,J＝7.6Hz,1H),8.38(d,J＝2.4Hz,1H),7.74(s,1H),7.66(dd,J＝4.0Hz,24.4Hz,1H),7.35(d,J＝12.8Hz,1H),6.68(d,J＝57.6Hz,1H), 5.78(s,1H),4.57(t,J＝5.6Hz,1H),4.45-4.22(m,6H),4.19-4.11(m,1H ),4.11-4.01(m,2H),3.82-3.68(m,2H),3.45(s,6H),3.37(d,J＝6.8Hz, 4H),3.22-3.12(m,2H),3.07-2.97(m,1H),2.88(s,4H),2.76-2.66(m,2 H),2.38(s,2H),2.26-2.13(m,2H),1.99-1.92(m,1H),1.86-1.74(m,1H ),1.69-1.61(m,1H),1.45(d,J＝6.4Hz,3H),1.34-1.23(m,1H),1.05-0.98(m,6H),0.95(s,3H),0.87(dd,J＝2.4Hz,6.4Hz,3H),0.57(s,3H)ppm.

Example 39

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 15F) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate L) replaces (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 39 (22.9 mg) was obtained as a yellow solid. MS calculated value 1145.5 (MH ⁺ ); measured value 1145.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.85-8.77(m,1H),8.69(d,J＝7.6Hz,1H),8.39(d,J＝2.8Hz,1H),7.7 2-7.64(m,2H),7.37-7.33(m,1H),7.09-6.93(m,1H),5.83-5.74(m,1H) ,4.46-4.37(m,1H),4.34-4.24(m,4H),4.18-4.14(m,1H),4.11-4.04(m ,1H),3.96-3.89(m,1H),3.80-3.71(m,2H),3.64-3.50(m,2H),3.45-3. 41(m,5H),3.36(s,3H),3.20-3.13(m,4H),3.04-2.98(m,1H),2.90-2.8 6(m,4H),2.83-2.78(m,1H),2.73-2.68(m,1H),2.25-2.16(m,2H),2.06 -2.01(m,2H),1.97-1.76(m,5H),1.68-1.53(m,3H),1.44(d,J＝6.4Hz,3 H),1.02-0.98(m,5H),0.95(s,3H),0.89-0.86(m,3H),0.57(s,3H)ppm.

Example 40

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

Similar to the preparation in Example 13, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 40 (26.2 mg) was obtained as a yellow solid. MS calculated value 10¹⁰.⁵ ( ^MH⁺ ); measured value 10¹⁰.⁴ ( ^MH⁺ ). ¹ H NMR (400MHz, methanol-d ₄ ) δ = 8.66 (t, J = 7.8Hz, 1H), 8.50 (d, J = 2.4Hz, 1H), 7.72 (br s,1H),7.63(t,J＝2.9Hz,1H),7.34(dd,J＝2.9,12.6Hz,1H),6.69-6.44(m,1H),5.83(br t,J＝7.5Hz,1H),5.01-4.92(m,1H),4.63-3.86(m,11H),3.84-3.65(m,3H),3.64-3.53(m,3H),3.45(br d,J＝15.2Hz,3H),3.36(br d,J＝3.4Hz,3H),3.31-3.22(m,3H),3.10-2.94(m,4H),2.81(br t,J＝12.4Hz,1H),2.65(br t,J＝13.4Hz,1H),2.52-2.37(m,1H),2.19(br d,J＝12.2Hz,1H),1.96(brd,J＝12.8Hz,1H),1.90-1.71(m,2H),1.70-1.54(m,1H),1.45(d,J＝6.2Hz,3H),1.17-0.77(m,12H),0.54(br d,J=9.3Hz,3H)ppm.

Example 41

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

Similar to the preparation in Example 13, using 3-[(2S)-2-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [C28C-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-tert-butyl ester (compound 40B) replacing 3-[(2R)-2-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared by reacting [-octadecano-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-3-methyl-butoxy]azacyclobutane-1-carboxylic acid tert-butyl ester (compound 13I). Example 41 (32.9 mg) was obtained as a yellow solid. MS calculated value 10¹⁰.⁵ ( ^MH⁺ ); measured value 10¹⁰.⁴ ( ^MH⁺ ). ^¹H NMR (400MHz, methanol- _d⁴ ) δ＝8.68(d,J＝7.4Hz,1H),8.51(d,J＝2.8Hz,1H),7.80(d,J＝2.8Hz,1H),7.65(d,J＝2.4Hz,1H),7.35(d,J＝12.6Hz,1H),6.75-6.53(m,1H),5.86(br) d,J＝7.1Hz,1H),5.03-4.92(m,1H),4.54-4.01(m,11H),4.01-3.83(m,2H),3 .81-3.66(m,3H),3.66-3.50(m,4H),3.50-3.38(m,3H),3.36(s,3H),3.02(br s,1H),2.99(s,3H),2.86-2.74(m,1H),2.74-2.62(m,1H),2.48-2.34(m,1H),2.17(br d,J＝12.1Hz,1H),2.00-1.85(m,2H),1.83-1.70(m,1H),1.68-1.55(m,1H),1.45(d,J＝6.3Hz,3H),1.09-0.85(m,12H),0.57(s,3H)ppm.

Similar to the preparation of compound 13J, it was prepared by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Compound 40B was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D).

Example 42

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 6, this was achieved by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^{22 ,26} ]octadec-1(25), ^2,5 (28), ^19,22(26 ),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 42 (7.8 mg) was obtained as a yellow solid. MS calculated value 1213.5 (MH ⁺ ); measured value 1213.4 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ＝8.70 (d, J＝7.6 Hz, 1H), 8.42 (d, J＝2.8 Hz, 1H), 7.74–7.67 (m, 1H), 7.52–7.41 (m, 2H), 7.13–6.87 (m, 1H), 5.75–5.65 (m, 1H), 5.20–5.11 (m, 1H), 4.81–4.78 (m, 1H), 4.48–4.34 (m, 1H), 4.25–4.05 (m, 3H), 3.98–3.65 (m, 3H),3.52-3.33(m,11H),3.30-3.09(m,6H),2.91-2.78(m,5H),2.68-2.57(m,1H),2.32-2.06(m,4H),2.00-1 .77(m,6H),1.70-1.57(m,3H),1.44(d,J＝6.0Hz,3H),1.03-0.92(m,6H),0.90-0.84(m,3H),0.47(s,3H)ppm.

Examples 43 and 45

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 43)

(Example 43)

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide (Example 45)

(Example 45)

Similar to the preparation in Example 4, this was achieved by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Example 43 was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26} ]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 43 (43.9 mg) was obtained as a yellow solid. MS calculated value 1185.4 (MH ⁺ ); measured value 1185.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ 8.82–8.71 (m, ¹H), 8.44 (br⁴). s,1H),7.84-7.66(m,1H),7.56-7.45(m,2H),6.96-6.71(m,1H),5.77-5.65(m,1H),5.26-5.16(m,1H),4.49 -4.33(m,2H),4.31-4.19(m,2H),4.07-3.69(m,5H),3.66-3.55(m,2H),3.54-3.34(m,10H),3.25-3.12(m,3H ),2.96-2.78(m,5H),2.72-2.57(m,1H),2.40-2.17(m,3H),2.16-1.92(m,4H),1.91-1.76(m,1H),1.75-1.5 7(m,1H),1.53-1.41(m,3H),1.38-1.14(m,1H),1.06-0.97(m,6H),0.94-0.86(m,3H),0.59-0.42(m,3H)ppm.

Similar to the preparation in Example 4, this was achieved by using (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13} .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate J) replaces (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0] Example ⁴⁵ was prepared by reacting octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 45 (24.2 mg) was obtained as a white solid. MS calculated value 1185.4 (MH ⁺ ); measured value 1185.5 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.70(d,J＝7.6Hz,1H),8.42(d,J＝2.8Hz,1H),7.71(t,J＝2.4Hz,1H),7.57(s,1H),7.48(d,J＝12.8Hz,1H),6.84(dd,J＝10.8Hz,48.8Hz,1H) ,5.72(s,1H),5.24-5.21(m,2H),4.48-4.18(m,3H),3.82-3.68(m,4H) ,3.68-3.55(m,2H),3.55-3.44(m,3H),3.42(d,J＝3.6Hz,4H),3.37(s, 3H),3.30-3.08(m,5H),2.89(t,J＝4.8Hz,4H),2.86-2.77(m,1H),2.64 (d,J＝14.4Hz,1H),2.32-2.15(m,3H),2.14-2.01(m,3H),1.98-1.90(m ,1H),1.88-1.74(m,1H),1.69-1.57(m,1H),1.45(d,J＝6.4Hz,3H),1.01(d,J＝6.4Hz,3H),0.99(s,3H),0.90(d,J＝6.4Hz,3H),0.49(s,3H)ppm.

Example 44

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

Similar to the preparation in Example 1, 1-tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate J) substituted for (S)-1-Boc-pyrrolidine-3-carboxylic acid (compound 1D) and (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E). Example 44 (18.4 mg) was obtained as a grayish-white solid. MS calculated value 1205.4 (MH ⁺ ); measured value 1205.6 (MH ⁺ ), ^1H NMR (400 MHz, methanol- _d4) )δ＝8.69(d,J＝7.2Hz,1H),8.42(d,J＝2.8Hz,1H),7.69(s,1H),7.58-7.51(m,1H),7.48(d,J＝12.4Hz,1H),7.11-6.97( m,1H),5.74-5.68(m,1H),5.24-5.10(m,2H),4.75(d,J＝11.2Hz,1H),4.46-4.34(m,2H),4.27-4.16(m,2H),4.00-3.88 (m,1H),3.81-3.69(m,2H),3.57-3.49(m,1H),3.47-3.38(m,5H),3.37(s,3H),3.22-3.17(m,4H),3.16-3.12(m,2H), 3.10-3.01(m,1H),2.88(t,J＝4.4Hz,5H),2.84-2.77(m,1H),2.68-2.59(m,1H),2.35-2.15(m,6H),1.99-1.93(m,1H). 1.88-1.77(m,1H),1.69-1.59(m,1H),1.45(d,J＝6.0Hz,3H),1.00-0.95(m,6H),0.89(d,J＝6.4Hz,3H),0.48(d,J＝5.6Hz,3H).

Example 46

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octane-6-yl)-3-methyl-butyric acid (compound 25H) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26} [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate J) replacing (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 46 (17.0 mg) was obtained as a grayish-white solid. MS calculated value 1171.4 (MH ⁺ ); measured value 1171.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d4) )δ＝8.72(d,J＝7.6Hz,1H),8.42(d,J＝2.4Hz,1H),7.74(d,J＝2.4Hz,1H),7.70(d,J＝2.0Hz,1H),7.50(d,J＝12.4Hz,1H),6.69(dd,J＝1.6Hz,48.8H z,1H),5.74-5.69(m,1H),4.44-4.21(m,5H),3.84-3.67(m,2H),3.59-3 .52(m,1H),3.46(s,6H),3.40(d,J＝1.2Hz,3H),3.24-3.14(m,3H),3.14 -3.08(m,1H),2.90(t,J＝5.2Hz,4H),2.85-2.77(m,1H),2.71-2.61(m,1 H),2.44-2.36(m,2H),2.27-2.14(m,2H),2.01-1.90(m,1H),1.90-1.71 (m,1H),1.70-1.55(m,1H),1.47(d,J＝6.4Hz,3H),1.25(s,2H),1.20(s,3H),1.02-0.96(m,6H),0.88(dd,J＝3.2Hz,6.4Hz,3H),0.51(s,3H)ppm.

Example 47

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 15F) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26} [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate J) replacing (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 47 (21.1 mg) was obtained as a white solid. MS calculated value 1199.5 (MH ⁺ ); measured value 1199.4 (MH ⁺ ), ^1H NMR (400 MHz, methanol- _d4) )δ＝8.86-8.79(m,1H),8.69(d,J＝7.2Hz,1H),8.44-8.40(m,1H),7.69(s,1 H),7.51-7.43(m,2H),7.12-6.90(m,1H),5.69(s,1H),5.22-5.11(m,1H), 4.44-4.37(m,1H),4.34-4.19(m,4H),3.99-3.87(m,1H),3.81-3.68(m,2H ),3.62-3.58(m,1H),3.49-3.42(m,2H),3.41-3.37(m,4H),3.35(s,3H),3. 28-3.23(m,1H),3.20-3.09(m,4H),2.90-2.86(m,4H),2.84-2.78(m,1H), 2.62(d,J＝14.4Hz,1H),2.26-2.19(m,2H),2.10-2.02(m,2H),2.00-1.90( m,2H),1.87-1.76(m,2H),1.70-1.53(m,3H),1.44(d,J＝6.0Hz,3H),1.18( t,J＝7.2Hz,1H),1.03-0.96(m,6H),0.88(d,J＝5.6Hz,3H),0.47(s,3H)ppm.

Example 48

(2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 6, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate D) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 48 (10 mg) was obtained as a yellow solid. MS calculated value 1159.5 (MH ⁺ ); measured value 1159.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ＝8.69(d, J＝7.6 Hz, 1H), 8.40(d, J＝2.8 Hz, 1H), 7.68–7.64(m, 1H), 7.63–7.57(m, 1H), 7.35(d, J＝12.8 Hz, 1H), 7.11–6.87(m, 1H), 5.82

-5.71(m,1H),4.82-4.75(m,2H),4.45-4.38(m,1H),4.36-4.29(m,1H),4.27

-4.03(m,4H),3.97-3.81(m,1H),3.79-3.70(m,2H),3.47-3.42(m,6H),3.36(s ,3H),3.30-3.12(m,5H),3.08-2.96(m,1H),2.92-2.79(m,5H),2.73-2.64(m,1 H),2.32-2.06(m,4H),2.00-1.90(m,1H),1.90-1.72(m,5H),1.68-1.55(m,3H) ,1.44(d,J＝6.4Hz,3H),1.05-0.94(m,9H),0.89-0.82(m,3H),0.55(s,3H)ppm.

Example 49

1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidine-4-carboxamide]

Similar to the preparation in Example 1, this was achieved by using 1-tert-butoxycarbonyl-4-fluoro-piperidine-4-carboxylic acid and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate L) substituted for (S)-1-Boc-pyrrolidine-3-carboxylic acid (compound 1D) and (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E). Example 49 (25.1 mg) was obtained as a yellow solid. MS calculated value 1151.5 (MH ⁺ ); measured value 1151.5 (MH ⁺ ), ^1H NMR (400 MHz, methanol- _d4) )δ＝8.70(d,J＝7.6Hz,1H),8.39(d,J＝2.8Hz,1H),7.85-7.78(m,1H),7.67 -7.60(m,1H),7.36(d,J＝12.4Hz,1H),7.11-6.94(d,J＝49.2Hz,1H),5.89 -5.69(m,1H),4.77-4.67(m,1H),4.46-4.32(m,3H),4.30-4.21(m,1H),4 .18-4.12(m,1H),4.10-4.00(m,1H),3.96-3.86(m,1H),3.81-3.71(m,2H) ,3.51-3.43(m,5H),3.43-3.34(m,4H),3.28-3.13(m,7H),3.03-2.98(m, 1H),2.92-2.87(m,4H),2.86-2.78(m,1H),2.76-2.66(m,1H),2.36-2.08( m,6H),1.99-1.90(m,1H),1.85-1.71(m,1H),1.70-1.58(m,1H),1.45(d, J＝6.4Hz,3H),1.04-0.93(m,9H),0.91-0.80(m,3H),0.65-1.53m,3H)ppm.

Example 50

(2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(8-tert-butoxycarbonyl-1-oxo-2,8-diazaspiro[4.5]decane-2-yl)-3-methyl-butyric acid (compound 15F) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate M) replaces (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 50 (51.8 mg) was obtained as a yellow solid. MS calculated value 1064.5 (MH ⁺ ); measured value 1064.3 (MH ⁺ ). ^1H NMR (400 MHz, chloroform-d) δ＝8.99(br s, 1H), 8.66-8.64(d, J＝7.5 Hz, 1H), 7.60-7.58(dd, J＝1.8, 8.4 Hz, 1H), 7.45-7.41(dd, J＝2.3, 16.1 Hz, 1H), 7.32-7.30(br d, J＝8.9 Hz, 0.5 H), 7.19-7.18(br d,J＝8.8Hz,0.5H),7.14-7.11(d,J＝12.3Hz,1H),6.57-6.54(m,0.5H),6.40-6.28(m,0.5H),5.81(td,J＝9 .2,13.9Hz,1H),4.66-4.52(m,1H),4.40(q,J＝6.4Hz,1H),4.31-4.22(m,1H),4.19-4.03(m,5H),3.92(br t,J＝4.5Hz,4H),3.83(br d,J＝11.2Hz,1H),3.69(br d,J＝11.1Hz,1H),3.62-3.54(m,8H),3.45-3.36(m,9H),3.21-3.07(m,2H),2.78-2.65(m,1H),2.50-2.30(m,2H),2.22(br d,J＝12.0Hz,1H),2.00-1.96(m,2H),1.92-1.76(m,1H),1.61(br d,J＝10.1Hz,2H),1.55-1.54(d,J＝6.4Hz,3H),1.06-0.93(m,9H),0.89-0.87(dd,J＝2.4,6.4Hz,3H),0.49(s,3H)ppm.

Examples 51 and 53

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide (Example 51)

(Example 51)

(2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide (Example 53)

(Example 53)

Similar to the preparation in Example 4, this was achieved by using (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate M) replaces (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0] Example ⁵¹ was prepared using octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 51 (21.4 mg) was obtained as a yellow solid. MS calculated value 1050.4 (MH ⁺ ); measured value 1050.4 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ＝8.70(d,J＝7.6Hz,1H),8.39(d,J＝2.8Hz,1H),7.89(dd,J＝2.8,9.6Hz,1H),7.65(s,1H),7.38(d,J＝12.8Hz,1H),8.90-6.74( m,1H),5.79(d,J＝9.2Hz,1H),4.44-4.37(m,2H),4.33-4.24(m,2H),4.18-4.12(m,1H),4.08-4.00(m,1H),3.87(t,J＝4.4Hz,4H ),3.81-3.70(m,4H),3.68–3.63(m,1H),3.58–3.54(m,1H),3.51-3.47(m,1H),3.45–3.41(m,5H),3.40(s,3H),3.29-3.23(m,1 H),3.02(d,J＝14.4Hz,1H),2.86-2.71(m,2H),2.28-2.15(m,3H),2.09-2.00(m,3H),1.98-1.93(m,1H),1.84-1.75(m,1H),1.68 -1.60(m,1H),1.46(d,J＝6.4Hz,3H),1.34-1.28(m,1H),1.04-0.99(m,6H),0.97(s,3H),0.88(d,J＝6.4Hz,3H),0.60(s,3H).

Similar to the preparation in Example 4, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 Example 53 was prepared by replacing (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadecane-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate D). Example 53 (28.8 mg) was obtained as a yellow solid. MS calculated value 1050.4 (MH ⁺ ); measured value 1050.5 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ＝8.88–8.87 (m, 1H), 8.72–8.68 (m, 1H), 8.38 (t, J＝2.4 Hz, 1H), 7.80 (t, J＝2.8 Hz, 1H), 7.74–7.64 (m, 1H), 7.39–7.34 (m, 1H), 6.91–6.69 (m, 1H), 5.81–5.73 (m, 1H), 4.45–4.36 (m, 2H), 4 .34-4.30(m,1H),4.29-4.23(m,1H),4.21-4.13(m,1H),4.11-4.02(m,1H),3.93-3.85(m,5H ),3.81-3.72(m,4H),3.63-3.51(m,3H),3.49-3.43(m,2H),3.42-3.38(m,7H),3.09-2.99(m 1H),2.87-2.78(m,1H),2.70(t,J＝14.0Hz,1H),2.33-2.25(m,1H),2.24-2.16(m,2H),2.12-2.06(m,2H),2.02-1.91(m,2H),1.85-1. 76(m,1H),1.70-1.61(m,1H),1.46(d,J＝6.4Hz,3H),1.04-0.98(m,6H),0.97-0.96(m,2H),0.89(d,J＝6.8Hz,3H),0.61–0.55(m,3H).

Example 52

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octane-6-yl)-3-methyl-butyric acid (compound 25H) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate M) replaces (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 52 (16.7 mg) was obtained as a yellow solid. MS calculated value 1036.4 (MH ⁺ ); measured value 1036.4 (MH ⁺ ). ^1H NMR (400 MHz, methanol- _d4) )δ=8.69(d,J=7.2Hz,1H),8.38(d,J=2.4Hz,1H),7.85-7.82(m,1H),7.69-7. 63(m,1H),7.37(d,J＝12.4Hz,1H),6.75-6.62(m,1H),5.80-5.74(m,1H),4.5 7(t,J＝10.0Hz,1H),4.41-4.34(m,3H),4.32-4.26(m,2H),4.19-4.13(m,1H) ,4.10-4.03(m,2H),3.87(t,J＝4.8Hz,4H),3.80-3.71(m,2H),3.53-3.46(m,2 H),3.42-3.41(m,3H),3.39-3.38(m,3H),3.07-2.98(m,1H),2.87-2.79(m,1 H),2.75-2.68(m,1H),2.41-2.35(m,2H),2.28-2.14(m,3H),2.07-1.90(m,2 H),1.85-1.76(m,1H),1.66-1.60(m,1H),1.46(d,J＝6.4Hz,3H),1.33-1.29( m,2H),1.03-0.98(m,6H),0.98-0.94(m,3H),0.89-0.85(m,3H),0.58(s,3H).

Example 54

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

Similar to the preparation in Example 1, this was achieved by using 1-tert-butoxycarbonyl-4-fluoro-piperidin-4-carboxylic acid and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate M) replaces (S)-1-Boc-pyrrolidine-3-carboxylic acid (compound 1D) and (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] The title compound was prepared from octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate E). Example 54 (44.2 mg) was obtained as a yellow solid. MS calculated value 1070.5 (MH ⁺ ); measured value 1070.3 (MH ⁺ ), ^1H NMR (400 MHz, methanol- _d4) )δ＝8.70(d,J＝7.2Hz,1H),8.39(d,J＝2.8Hz,1H),7.90-7.83(m,1H),7.64(d,J＝2.4Hz,1H),7.37(d,J＝12.8Hz,1H),7.12-6.95(d,J＝49.2Hz,1H) ,5.87-5.72(m,1H),4.85-4.69(m,1H),4.46-4.22(m,4H),4.19-4.12(m ,1H),4.10-4.00(m,1H),3.98-3.90(m,1H),3.88(t,J＝4.8Hz,4H),3.81- 3.71(m,2H),3.55-3.47(m,1H),3.45-3.38(m,8H),3.30-3.20(m,2H),3 .20-3.13(m,3H),3.06-2.98(m,1H),2.89-2.67(m,2H),2.35-2.07(m,6 H),1.99-1.90(m,1H),1.86-1.73(m,1H),1.72-1.57(m,1H),1.46(d,J＝ 6.4Hz, 3H), 1.05-0.94 (m, 9H), 0.88 (d, J = 6.4Hz, 3H), 0.66-0.55 (m, 3H).

Example 55

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

Similar to the preparation in Example 1, this was achieved by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 The title compound was prepared by replacing (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 2,5 .1 9,13 .0 ^22,26 ]octadec-1(25), ^2,5 (28), ^19,22 (26),23-hexane-8,14-dione (intermediate E) with octadec-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate E). Example 55 (24.7 mg) was obtained as a yellow solid. MS calculated value: 1038.4 (MH ⁺ ); measured value: 1038.6 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ＝8.73–8.68 (m, 1H), 8.41–8.36 (m, 1H), 7.91–7.82 (m, 1H), 7.70–7.67 (m, 1H), 7.37 (d, J＝16.0 Hz, 1H), 6.92–6.72 (m, 1H), 5.81–5.70 (m, 1H), 4.81–4.71 (dd, J＝4.0, 12.0 Hz, 1H) ,4.47-4.33(m,2H),4.32-4.22(m,1H),4.21-4.11(m,1H),4.09-4.00(m,1H),3.97-3.85 (m,5H),3.82-3.72(m,4H),3.69-3.54(m,2H),3.46-3.37(m,8H),3.28-3.21(m,1H),3.15 -3.05(m,3H),3.04-2.95(m,1H),2.88-2.67(m,2H),2.41-2.06(m,4H),1.97-1.57(m,3 H), 1.46 (d, J = 8.0Hz, 3H), 1.08-0.93 (m, 9H), 0.86 (d, J = 4.0Hz, 3H), 0.73-0.56 (m, 3H).

Example 57

(2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butanamide]

Similar to the preparation in Example 4, this was achieved by using (2S)-2-[(5R)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 1) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate I) replaces (2S)-2-[(5S)-7-tert-butoxycarbonyl-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-3-methyl-butyric acid (compound 4f 2) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9, 13]} Example ⁵⁷ was prepared using octadecano-1(25),2,5(28),19,22(26),23-hexaden-8,14-dione (intermediate D). Example 57 (26 mg) was obtained as a white solid. MS calculated value 1104.4 (MH ⁺ ); measured value 1104.4 (MH ⁺ ). ^¹H NMR (400MHz, methanol- _d⁴ ) δ＝8.86-8.98(d,J＝7.6Hz,1H), 8.66-8.75(d,J＝7.2Hz,1H), 8.32-8.49(d,J＝2.8Hz,1H), 7.66-7.75(t,J＝2.8Hz,1H), 7.56-7.63(m,1H), 7.40-7.54(d,J＝12.4Hz,1H), 6.73-6.94(dd, _J₁ ＝11.2Hz,J₂ ₎ ＝49.2Hz,1H),5.60-5.81(t,J＝8.4Hz,1H),5.11-5.31(m,1H),4.38-4.47(d,J＝12.4Hz,1H),4.30-4.37(dd,J ₁ ＝3.2Hz,J ₂ ＝11.2Hz,1H),4.16-4.30(m,2H),3.82-4.00(m,4H),3.65-3.82(m,5H),3.55-3.64(m,2H),3.4 2-3.54(m,3H),3.33-3.40(m,7H),3.07-3.18(m,1H),2.76-2.90(m,1H),2.58-2.74(m,1H),2. 16-2.33(m,3H),2.02-2.14(m,3H),1.89-2.00(m,2H),1.75-1.88(m,1H),1.56-1.71(m,1H),1 .41-1.52(d,J＝6.4Hz,3H),0.94-1.07(m,6H),0.86-0.92(d,J＝6.4Hz,3H),0.43-0.54(s,3H).

Example 58

(2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide

Similar to the preparation in Example 6, this was achieved by using (2S)-2-(2-tert-butoxycarbonyl-5-oxo-2,6-diazaspiro[3.4]octane-6-yl)-3-methyl-butyric acid (compound 25H) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-8,14-dione (intermediate I) replaces (2S)-2-(9-tert-butoxycarbonyl-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl)-3-methyl-butyric acid (compound 6J) and (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] The title compound was prepared from octadecano-1 (25), 2, 5 (28), 19, 22 (26), 23-hexaden-8, 14-dione (intermediate D). Example 58 (24.0 mg) was obtained as a white solid. MS calculated value 1090.4 (MH ⁺ ); measured value 1090.4 (MH ⁺ ). ^¹H NMR (400 MHz, methanol- _d⁴ ) δ＝8.76–8.91 (d, J＝8.4 Hz, 1H), 8.56–8.64 (d, J＝7.6 Hz, 1H), 8.28–8.36 (d, J＝2.4 Hz, 1H), 7.57–7.67 (dd, J＝2.4 Hz, 17.6 Hz, 1H), 7.44–7.52 (m, 1H), 7.35–7.42 (d, J＝12.8 Hz). z,1H),6.51-6.68(dd,J＝3.2Hz,52.4Hz,1H),5.55-5.70(t,J＝8.4Hz,1H),4.99-5.21 (m,1H),4.43-4.56(m,1H),4.08-4.38(m,6H),3.91-4.04(m,1H),3.73-3.84(t,J＝4.8 Hz,4H),3.66-3.72(m,1H),3.56-3.64(m,1H),3.42-3.50(m,1H),3.32-3.41(m,2H), 3.23-3.30(m,8H),2.97-3.07(m,1H),2.65-2.78(m,1H),2.49-2.58(m,1H),2.24-2. 35(m,2H),2.06-2.19(m,2H),1.80-1.91(m,1H),1.64-1.79(m,1H),1.45-1.61(m,1H ), 1.31-1.39 (d, J = 8Hz, 3H), 0.85-0.97 (m, 6H), 0.73-0.81 (m, 3H), 0.34-0.44 (s, 3H).

Biological examples

Compounds RM461 (page 115 of Figure 1) from WO2020132597, RM351 (page 88 of Figure 1) from WO2020132597, and RM44 (page 11 of Figure 1) from WO2020132597 are referenced as reference compounds in this invention.

The applicant, following the procedure described in WO2020132597, further synthesized RM461-A and RM44-A as analogs of RM461 and RM44, respectively, by replacing chloroacetamide with 2-chloro-2-fluoroacetamide, and provided comparative data to demonstrate the improvement and illustrate the technical problem solved herein.

Example 59

GSH Reactivity

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

Example <![CDATA[T_1/2(hours) in GSH]]> Residual amount (%) of GSH at 6 hours RM461 5 44 RM461-A Stablize 100 RM351 4.3 37 RM44 8.9 60 RM44-A Stablize 100 Example 37 Stablize 100

The above results clearly demonstrate that the reference compounds (RM461, RM351, and RM44) 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 the reference compounds RM461 and RM44 with their analogues RM461-A and RM44-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 60

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) supplemented with Williams' E medium containing 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 2. Human hepatocyte stability of examples and compounds of the present invention

The above results clearly demonstrate that, in human hepatocyte stability assays, the reference compound (RM44) exhibited moderate clearance, while Example 37 of the present invention maintained low clearance. In particular, comparing the reference compound RM44 with its analogue RM44-A, the only difference being the substitution of chloroacetamide with 2-chloro-2-fluoroacetamide, as with the compounds of the present invention, it is surprising that such a small change could achieve a reduction in clearance in human hepatocyte stability assays.

Achieving low clearance rates can improve the in vivo performance of compounds, such as dose reduction, enhanced exposure, and prolonged half-life.

Example 61

Cell viability assay

The purpose of this cell assay was to determine the effect of the test 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 Count 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.

Compared with reference compounds RM461 and RM351, the compounds of the present invention clearly demonstrate significantly improved overall cell potency in G12C, G12D and G12V.

Table 3. Examples of the present invention and the activity of compounds in KRAS cell viability assays

Example 62

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

Example 63

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

In this example, TR-FRET was also used to measure the compound- or compound-CYPA-dependent disruption of the KRAS G12C-BRAF complex. This protocol was also used to measure the disruption of KRAS G12D or KRAS G12V binding to BRAF by the compounds of this invention, respectively. 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 in a 16-point 3-fold dilution series and incubated for 3 hours. Then, mixtures of MAb anti-6His-XL665 (Cisbio, 61HISXLB) and Mab anti-GST-TB well compound (Cisbio, 61GSTTLB) were added at final concentrations of 6.67 nM and 0.21 nM, respectively, and the plates were incubated for another 1.5 hours. TR-FRET signals were read on a PHERstar FSX microplate reader (Ex 320 nm, Em 665/615 nm). Compounds that promoted the disruption of the KRAS-BRAF complex were identified as those causing a decrease in TR-FRET ratios compared to the DMSO control wells. Table 5. Examples of the invention and the activity of compounds in the KRAS-BRAF/CYPA (500 nM) interaction assay.

Example 64

pERK inhibition assay

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

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

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

Table 6. Examples of the present invention and the activity of compounds in KRAS pERK inhibition assays

Claims (33)

1. A compound of formula (I), in ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 5-oxo-2,6-diazaspiro[3.4]octyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decyl substituted with (dihalo _-C1-6 alkyl)carbonyl, or Azahexacyclic butyloxy _C1-6 alkyl groups substituted with (dihalo- _C1-6 alkyl)carbonyl groups; ^R8 is a _C1-6 alkyl group; R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _-C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _-C1-6 alkyl) carbonyl group; ^R2 is a _C1-6 alkyl group; ^R3 is H, _C1-6 alkyl, or halogen; ^R4 is H or halogen; R ⁵ is a _C1-6 alkyl or a halo- _C1-6 alkyl; ^R6 is a _C1-6 alkoxy- _C1-6 alkyl group; R ⁷ is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino; ^A1 is an imidazolyl group or a hydroxylated phenylene group; ^A2 is a _C1-6 alkylene group; The condition is that ^R3 and ^R4 are not both H at the same time; Or its medicinal salt. 2. A compound of formula (Ia), in ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 5-oxo-2,6-diazaspiro[3.4]octyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 5-oxo-2,6-diazaspiro[3.5]nonyl substituted with (dihalo _-C1-6 alkyl)carbonyl, 6-oxo-2,7-diazaspiro[4.5]decyl substituted with (dihalo _-C1-6 alkyl)carbonyl, or Azahexacyclic butyloxy _C1-6 alkyl groups substituted with (dihalo- _C1-6 alkyl)carbonyl groups; ^R8 is a _C1-6 alkyl group; R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _C1-6 alkyl) carbonyl group; ^R2 is a _C1-6 alkyl group; ^R3 is H, _C1-6 alkyl, or halogen; ^R4 is H or halogen; R ⁵ is a _C1-6 alkyl or a halo- _C1-6 alkyl; ^R6 is a _C1-6 alkoxy- _C1-6 alkyl group; R ⁷ is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino; ^A1 is an imidazolyl group or a hydroxylated phenylene group; ^A2 is a _C1-6 alkylene group; The condition is that ^R3 and ^R4 are not both H at the same time; Or its medicinal salt. 3. The compound according to claim 1 or 2, wherein... ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, or 5-oxo-2,6-diazaspiro[3,4]octyl groups substituted with (dihalo _-C1-6 alkyl)carbonyl groups; ^R8 is a _C1-6 alkyl group; R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _-C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _-C1-6 alkyl) carbonyl 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, 1-oxo-2,8-diazaspiro[4.5]decane-2-yl substituted with chloro(fluoro)acetyl, 1-oxo-2,9-diazaspiro[5.5]undecane-2-yl substituted with chloro(fluoro)acetyl, or 5-oxo-2,6-diazaspiro[3.4]octane-6-yl substituted with chloro(fluoro)acetyl; ^R8 is a methyl group; R ⁹ is a piperidinyl group that has been substituted twice with fluorine and chloro(fluoro)acetyl groups, or a pyrroleyl group that has been substituted with chloro(fluoro)acetyl groups. 5. The compound according to any one of claims 1 to 4, wherein ^R1 is (5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, (5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, 8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl, 9 -[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl, 2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl, 1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-4-piperidinyl or 1-[(2R)-2-chloro-2-fluoro-acetyl]pyrrolidine-3-yl. 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 H, methyl or fluorine. 8. The compound according to any one of claims 1 to 7, wherein ^R3 is fluorine. 9. The compound according to any one of claims 1 to 8, wherein ^R4 is H or fluorine. 10. The compound according to any one of claims 1 to 9, wherein ^R4 is H. 11. The compound according to any one of claims 1 to 10, wherein ^R5 is ethyl or 2,2,2-trifluoroethyl. 12. The compound according to any one of claims 1 to 11, wherein R ⁶ is 1-methoxyethyl. 13. The compound according to any one of claims 1 to 12, wherein R ⁷ is morpholino, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or 4-methylpiperazin-1-yl. 14. The compound according to any one of claims 1 to 13, wherein ^A1 is a compound in which bond “a” is attached to an indole ring. 15. The compound according to any one of claims 1 to 14, wherein ^A2 is a dimethylmethylene group. 16. The compound according to claim 1 or 2, wherein ^R1 is 1-oxo-2,7-diazaspiro[4.4]nonyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,8-diazaspiro[4.5]decyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, 1-oxo-2,9-diazaspiro[5.5]undecyl group substituted with (dihalo _-C1-6 alkyl)carbonyl group, or 5-oxo-2,6-diazaspiro[3,4]octyl group substituted with (dihalo _-C1-6 alkyl)carbonyl; ^R8 is a _C1-6 alkyl group; R ⁹ is a piperidinyl group that has been substituted twice with a halogen and a (dihalo _C1-6 alkyl) carbonyl group, or a pyrrolidinyl group that has been substituted with a (dihalo _C1-6 alkyl) carbonyl group; ^R2 is a _C1-6 alkyl group; ^R3 is a halogen; ^R4 is H; R ⁵ is a _C1-6 alkyl or a halo- _C1-6 alkyl; ^R6 is a _C1-6 alkoxy- _C1-6 alkyl group; R ⁷ is morpholino, (halogenated _C1-6 alkyl)piperazino, or _C1-6 alkylpiperazino; ^A1 is where the "a" bond is attached to the indole ring; ^A2 is a _C1-6 alkylene group; Or its medicinal salt. 17. The compound according to claim 16, wherein... ^R1 is (5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, (5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl, 8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl, 9 -[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl, 2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl, 1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-4-piperidinyl or 1-[(2R)-2-chloro-2-fluoro-acetyl]pyrrolidine-3-yl; ^R2 is isopropyl; ^R3 is fluorine; ^R4 is H; R ⁵ is ethyl or 2,2,2-trifluoroethyl; R ⁶ is (1S)-1-methoxyethyl; R ⁷ is morpholino, 4-(2,2,2-trifluoroethyl)piperazin-1-yl or 4-methylpiperazin-1-yl; ^A1 is where the "a" bond is attached to the indole ring; ^A2 is a dimethylmethylene group; Or its medicinal salt. 18. A compound selected from: (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26]} [Octadecacarbon-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)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26]} [28-octadec-1(25),2,5(28),19,22(26),23-hexene-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]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17,24-trimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [28-octadec-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(8S,14S)-25-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27] [26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; 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]-17,17,24-trimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide; 1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^{22 ,26]} [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide; (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.5]nonane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2R)-2-[[1-[(2R)-2-chloro-2-fluoro-acetyl]azacyclobutane-3-yl]oxymethyl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo-[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butanamide; (2S)-2-[[1-[(2R)-2-chloro-2-fluoro-acetyl]azacyclobutane-3-yl]oxymethyl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butanamide; (2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[(5S)-2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[(5R)-2-[(2R)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[(5R)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[(5S)-2-[(2S)-2-chloro-2-fluoro-acetyl]-6-oxo-2,7-diazaspiro[4.5]decane-7-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecano-1(25),2,5(28),19,22(26),23-hexane-7-yl]-3-methyl-butyramide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [Octadecacarbon-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]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 [Octadecacarbon-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-[[(8S,14S)-26-fluoro-4-hydroxy-(21M)-21-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-18,18-dimethyl-9,15-dioxo-22-(2,2,2-trifluoroethyl)-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 ^2,6 .1 ^10,14 .0 ^23,27] [26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-pyrrolidine-3-carboxamide; 1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidine-4-carboxamide; (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2R)-2-[[1-[(2R)-2-chloro-2-fluoro-acetyl]azacyclobutane-3-yl]oxymethyl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butanamide; (2S)-2-[[1-[(2R)-2-chloro-2-fluoro-acetyl]azacyclobutane-3-yl]oxymethyl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butanamide; (2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; 1-[(2R)-2-chloro-2-fluoro-acetyl]-4-fluoro-N-[(1S)-1-[[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2 ,5} .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-N-methyl-piperidine-4-carboxamide; (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; (2S)-2-[9-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,9-diazaspiro[5.5]undecane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]-3-methyl-butyramide; 1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ] [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexene-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidine-4-carboxamide; (2S)-2-[8-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,8-diazaspiro[4.5]decane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[(5S)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2, 5} .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[(5R)-7-[(2R)-2-chloro-2-fluoro-acetyl]-1-oxo-2,7-diazaspiro[4.4]nonane-2-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^{2, 5} .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^{9 ,13} .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; 1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexane-7-yl]carbamoyl]-2-methyl-propyl]-4-fluoro-N-methyl-piperidine-4-carboxamide; (3S)-1-[(2R)-2-chloro-2-fluoro-acetyl]-N-[(1S)-1-[[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-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)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 [Octadecacarbon-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methylbutyramide; and (2S)-2-[2-[(2R)-2-chloro-2-fluoro-acetyl]-5-oxo-2,6-diazaspiro[3.4]octane-6-yl]-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1 ^2,5 .1 ^9,13 .0 ^22,26 ]octadec-1(25),2,5(28),19,22(26),23-hexaden-7-yl]-3-methyl-butyramide; Or its medicinal salt. 19. A compound having the following structure: 20. A method for preparing the compound according to any one of claims 1 to 19, the method comprising any one of the following steps: a) In the presence of a coupling agent and a base, compound (II) is formed. It undergoes a coupling reaction with acid (III) to form the compound of formula (I); b) In the presence of a coupling agent and a base, compound of formula (VI) is... It undergoes a coupling reaction with acid (VII) to form compound (VIII). c) To make compound (XI) in the presence of a coupling agent and a base. It undergoes a coupling reaction with acid (XII) to form a compound of formula (XIII). Wherein Q is an unsubstituted or substituted piperidinyl or pyrrolidine; T is a dihalogenated _C1-6 alkyl; ^R1 , R2, ^R3 , ^R4 , ^R5 , ^R6 , ^R7 , ^A1 and ^A2 are as defined in any one of claims ¹ to 17; the coupling agent in steps a) to c) is _T3P , HATU, PyBOP or EDCI/HOBt; the base in steps a) to c) is TEA, DIEPA or DMAP. 21. The compound or pharmaceutical salt according to any one of claims 1 to 19, which is used as a therapeutically active substance. 22. A pharmaceutical composition comprising: a compound according to any one of claims 1 to 19; and a pharmaceutical excipient. 23. Use of the compound according to any one of claims 1 to 19 for the treatment of KRAS G12C protein-related diseases. 24. Use of the compound according to any one of claims 1 to 19 for the treatment of diseases related to KRASG12C, G12D and G12V proteins. 25. Use of the compound according to any one of claims 1 to 19 for suppressing the interaction between RAS and downstream effectors, wherein the downstream effectors are RAF and PI3K. 26. Use of the compound according to any one of claims 1 to 19 for inhibiting the transmission of carcinogenic MAPK and PI3K signaling. 27. Use of the compound according to any one of claims 1 to 19 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. 28. Use of the compound according to any one of claims 1 to 19 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. 29. The compound or pharmaceutical salt according to any one of claims 1 to 19, 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. 30. Use of the compound according to any one of claims 1 to 19 for the preparation of a medicament for the treatment or prevention of KRAS mutation-driven cancer, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer, and non-small cell lung cancer. 31. 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 19. 32. The compound or pharmaceutical salt according to any one of claims 1 to 19, prepared according to the method of claim 20. 33. The present invention as described in the specification.