CN120757571A

CN120757571A - Preparation and application of pyrimidothiopyranedione inhibitors of KRAS G12C mutant protein

Info

Publication number: CN120757571A
Application number: CN202511023003.1A
Authority: CN
Inventors: 梁永宏
Original assignee: Yaoya Technology Shanghai Co ltd
Current assignee: Yaoya Technology Shanghai Co ltd
Priority date: 2024-02-01
Filing date: 2024-02-01
Publication date: 2025-10-10
Also published as: CN118005656B; TW202532417A; WO2025162135A1; CN118005656A; TWI907258B

Abstract

本发明涉及KRAS G12C突变蛋白嘧啶并噻喃二酮类抑制剂及其用途，具体地而言本发明提供一种式(I)所示化合物，式中各取代在的定义如说明书中所述。此外还涉及该抑剂的组合物及其应用。本发明化合物具有良好的抑制肿瘤增长的活性。并具有良好的安全性。The present invention relates to KRAS G12C mutant protein pyrimidothiopyranedione inhibitors and their uses. Specifically, the present invention provides a compound represented by formula (I), wherein the substituents are defined as described in the specification. Furthermore, the present invention relates to compositions of the inhibitor and their uses. The compounds of the present invention have excellent activity in inhibiting tumor growth and have a good safety profile.

Description

Preparation and application of KRAS G12C mutant protein pyrimidothiopyrandione inhibitor

Technical Field

The invention belongs to the field of drug synthesis, and particularly relates to a novel KRAS G12C inhibitor, and a preparation method and application thereof.

Background

The present invention relates generally to novel compounds, methods for their preparation and use as KRAS G12C inhibitors (e.g. for the treatment of cancer).

RAS represents a closely related group of monomeric globular proteins of 189 amino acids (molecular weight 21 kDa) that are associated with the plasma membrane and bind GDP or GTPoRAS as molecular switches. When the RAS contains bound GDP, it is in a quiescent or off state, and in an "inactive state". In response to exposure of the cells to certain growth-promoting stimuli, the RAS is induced to convert its bound GDP to GTP. After binding to GTP, the RAS is "turned on" and is able to interact with and activate other proteins (its "downstream targets"). RAS proteins themselves have very low intrinsic ability to hydrolyze GTP back to GDP, thus placing themselves in an off state. Turning off the RAS requires an external protein called GTPase Activating Proteins (GAPs), which interact with the RAS and greatly accelerate the conversion of GTP to GDP. Any mutation in the RAS that affects its ability to interact with GAP or convert GTP back to GDP will result in an extended activation time of the protein, thus resulting in an extended cell signal that allows it to continue to grow and divide. Since these signals lead to cell growth and division, the hyperactive RAS signals may ultimately lead to cancer.

Structurally, the RAS protein comprises a G domain responsible for enzymatic activity of the RAS-guanosine nucleotidic binding and hydrolysis (GTPase reaction). It also contains a C-terminal extension called a CAAX box, which can be post-translationally modified and is responsible for targeting proteins to the membrane. The G domain is about 21-25kDa in size and comprises a phosphate binding ring (P-ring). The P-loop is the pocket in which the nucleic acid binds in the protein, which is a rigid part of the domain with conserved amino acid residues ((glycine 12, threonine 26 and lysine 16)) that is essential for nucleic acid binding and hydrolysis. The G domain also contains so-called Switch I (residues 30-40) and Switch II (residues 60-76) regions, both of which are dynamic parts of the protein, which are commonly referred to as "spring loaded" mechanisms as they are capable of switching between resting and loaded states. The key interaction is the hydrogen bond formed by threonine 35 and glycine 60, the Y-phosphate with GTP, which keeps the Switch1 and Switch2 regions in their active conformation, respectively. After GTP hydrolyses and releases phosphate, the two relax to an inactive GDP conformation.

The most well known members of the RAS subfamily are HRAS, KRAS and NRAS, mainly because of their association with multiple types of cancer. Any mutation in any of the three major isoforms of the RAS (HRAS, NRAS or KRAS) gene is the most common in human tumorigenesis. About 30% of human tumors were found to carry RAS gene mutations o notably, KRAS mutations were detected in 25-30% of tumors. In contrast, the oncogenic mutation rates occurring in NRAS and HRAS family members are much lower (8% and 3%, respectively). The most common KRAS mutations were found at residues G12 and G13 and residue Q61 of the P loop. G12C is a frequent mutation of the KRAS gene (glycine 12 to cysteine). Such mutations have been found in about 13% of the occurrences of cancer, about 43% of the occurrences of lung cancer, and about 100% of MYH-related polyposis (familial colon cancer syndrome).

As a leading edge target, KRAS G12C muteins have received great attention. Araxes (subsidiary of WELLSPRING) developed ARS-853 and ARST620 compounds in 2013 and 2016, respectively. In recent years, several patents have been filed for KRASG12C inhibitors, such as W02016164675 and W02016168540, which show good cell viability but poor pharmacokinetic properties for MRS-853 compounds, which are unsuitable for evaluating pharmacodynamics of animal models in vivo. Ars-1620 has high efficiency and selectivity for KRASG12C, and can achieve rapid and sustained targeting in vivo, thereby inducing tumor regression. The in vivo evidence provided by this study suggests that ARS-1620 represents a new generation of KRASG 12C-specific inhibitors with great therapeutic potential. WELLSPRING announces that FDA approved IND applications for ARS-3248. Other candidate KRAS ^G12C inhibitors include MRTX-849 from Mirati and BI-2852 from Boehringer Ingelheim, among others. Thus, despite advances in this field, there remains a need in the art for improved compounds and methods of treating cancer, for example, by inhibiting KRAS, HRAS or NRAS. The present invention meets this need and provides other related advantages.

Briefly, the present invention provides compounds, including stereoisomers, pharmaceutically acceptable salts, tautomers and prodrugs thereof, capable of modulating G12C mutant KRAS, HRAS and/or NRAS proteins. In some cases, the compound acts as an electrophile capable of forming a covalent bond with a cysteine residue at the 12 position of a KRAS, HRAS or NRAS G12C mutein. Methods of using such compounds for treating various diseases or conditions, such as cancer, are also provided.

Disclosure of Invention

A compound having the general formula (I), a stereoisomer, a pharmaceutically acceptable salt, a polymorph or an isomer thereof, wherein the compound of the general formula (I) has the structure:

Wherein, the

Each X ₁ is independently selected at each occurrence from N, CR ₄;

L ₁ is independently selected from the group consisting of-C _0-4 alkyl-, -CR ₈R₉-、-C_1-2 alkyl (R⁸)(OH)-、-C(O)-、-CR₈R₉O-、-OCR₈R₉-、-SCR₈R₉-、-CR₈R₉S-、-NR₈-、-NR₈C(O)-、-C(O)NR₈-、-NR₈C(O)NR₉-、-CF₂-、-O-、-S-、-S(O)_m-、-NR₈S(O)_m-、-S(O)_mNR₈-;

R ₁ is independently selected from C _3-8 cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spirocyclyl, 5-12 membered spiroheterocyclyl wherein said cycloalkyl, heterocycloalkyl, spirocyclyl, fused heterocyclyl, spiroheterocyclyl are substituted with one or more G ¹;

R ₄ is independently selected from H, D, cyano, halogen, C _1-6 alkyl, COOH, NHCOH, CONH ₂, OH, or NH ₂;

u is independently selected from-C _0-4 alkyl-, -CR ₈R₉-、-C_1-2 alkyl (R⁸)(OH)-、-C(O)-、-CR₈R₉O-、-OCR₈R₉-、-SCR₈R₉-、-CR₈R₉S-、-NR₈-、-NR₈C(O)-、-C(O)NR₈-、-NR₈C(O)NR₉-、-CF₂-、-O-、-S-、-S(O)_m-、-NR₈S(O)_m-、-S(O)_mNR₈-;

Y is absent or selected from C _3-8 cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spiroheterocyclyl, aryl or heteroaryl, wherein said cycloalkyl, heterocycloalkyl, spiroheterocyclyl, fused ring, fused heterocyclyl, spiroheterocyclyl, aryl or heteroaryl is optionally substituted with one or more G ²;

z is independently selected from cyano, -NR ₁₀ CN,

Bond c is a double bond or a triple bond;

When C is a double bond, R ^a、R^b and R ^c are each independently selected from H, deuterium, cyano, halogen, C _1-6 alkyl, C _3-6 cycloalkyl or 3-6 membered heterocyclyl. Wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted with 1 or more G ³;

R ^a and R ^b or R ^b and R ^c together with the carbon atom to which they are attached optionally form a 3-to 6-membered ring optionally containing heteroatoms;

When bond C is a triple bond, R ^a and R ^c are absent, R ^b is independently selected from H, deuterium, cyano, halogen, C _1-6 alkyl, C _3-6 cycloalkyl, or 3-6 membered heterocyclyl substituted with one or more G ⁴;

R ₁₀ is independently selected from H, deuterium, C _1-6 alkyl, C _3-6 cycloalkyl, or a 3-6 membered heterocyclyl, wherein the alkyl, cycloalkyl, and heterocyclyl are optionally substituted with 1 or more G ⁵;

Each Ar is independently selected from the group consisting of a 5-12 membered heteroaryl group at each occurrence, the heteroaryl group independently comprising 1,2, 3, or 4 heteroatoms selected from N, O, or S at each occurrence, wherein the heteroaryl group is optionally substituted with one or more G ⁶;

Each of G ¹、G²、G³、G⁴、G⁵ and G ⁶ is independently selected from deuterium, cyano, halogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl or 3-8 membered heterocyclyl, C _6-10 aryl, 5-10 membered heteroaryl 、-OR₁₁、-OC(O)NR₁₁R₁₂、-C(O)OR₁₁、-C(O)NR₁₁R₁₂、-C(O)R₁₁、-NR₁₁R₁₂、-NR₁₁C(O)R₁₂、-NR₁₁C(O)NR₁₂R₁₃、-S(O)_mR₁₁ or-NR ₁₁S(O)_mR₁₂, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl is optionally substituted with 1 or more deuterium, cyano, halogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl or 3-8 membered heterocyclyl, C _6-10 aryl, 5-10 membered heteroaryl 、-OR₁₄、-OC(O)NR₁₄R₁₅、-C(O)OR₁₄、-C(O)NR₁₄R₁₅、-C(O)R₁₄、-NR₁₄R₁₅、-NR₁₄C(O)R₁₅、-NR₁₄C(O)NR₁₅R₁₆、-S(O)_mR₁₄ or-NR ₁₄S(O)_nR₁₅ substituents;

R ⁸、R⁹、R¹¹、R¹²、R¹³、R¹⁴ and R ¹⁵ are each independently selected from hydrogen, deuterium, cyano, halogen, C _1-6 alkyl, C _3-8 cycloalkyl or 3-8 membered monocyclic heterocyclyl, monocyclic heteroaryl or phenyl;

And m, n is 1 or 2.

In some embodiments, the compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is further represented by formula (II-A), (II-B), (II-C), or (II-D)

In some embodiments, the compound of formula (I) or an isomer, solvate or precursor thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds, isomers, solvates or precursors thereof, or pharmaceutically acceptable salts thereof:

1. A compound having the general formula (I), a stereoisomer, a pharmaceutically acceptable salt, a polymorph or an isomer thereof, wherein the compound of the general formula (I) has the structure:

Wherein, the

Each X ₁ is independently selected at each occurrence from N, CR ₄;

z is independently selected from cyano, -NR ₁₀ CN,

Bond c is a double bond or a triple bond;

And m, n is 1 or 2.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is represented by the general formula (II-a), (II-B), (II-C), or (II-D):

3. the compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein Ar is selected from:

a compound according to claim 1, or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, and mixtures thereof, selected from the group consisting of:

In some embodiments, the compound of formula (I) or a stereoisomer, solvate, or precursor thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds, isomers, solvates, or precursors thereof, or pharmaceutically acceptable salts thereof:

In another aspect, the present invention also provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant.

In another aspect, the invention relates to a method of treating a KRAS G12C-associated disease in a mammal comprising administering to a mammal, preferably a human, in need of such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

In another aspect, the present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the prevention or treatment of KRAS G12C-related diseases.

In another aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating KRAS G12C-related diseases.

Detailed description of the preferred embodiments

The invention also provides a method for preparing the compound. The preparation of the compounds of the general formula (I) according to the invention can be carried out by the following exemplary methods and examples, which, however, should not be regarded as limiting the scope of the invention in any way. The compounds of the present invention may also be synthesized by synthetic techniques known to those skilled in the art, or by a combination of methods known in the art and methods described herein. The product obtained in each step is obtained using separation techniques known in the art including, but not limited to, extraction, filtration, distillation, crystallization, chromatographic separation, and the like. The starting materials and chemical reagents required for the synthesis may be conventionally synthesized or purchased according to literature (reaxys).

Unless otherwise indicated, temperatures are degrees celsius. Reagents were purchased from commercial suppliers of phase contrast medicine or micin, and these reagents were used directly without further purification unless otherwise indicated.

Unless otherwise indicated, the following reactions were carried out at room temperature, in anhydrous solvents, under positive pressure of nitrogen or gas or using a drying tube, glassware drying and/or heat drying.

Column chromatography purification was performed using 200-300 mesh silica gel from Qingdao ocean chemical plant, thin layer chromatography was performed using a thin layer chromatography silica gel prefabricated plate (HSGF 254) manufactured by Kagaku Co., ltd.) and MS was measured using a Therno LCD Fleet-type (ESI) liquid chromatography-mass spectrometer.

Nuclear magnetic data (1H NMR) Using a Bruker Avance-400MHz or Varian Oxford-400Hz nuclear magnetic instrument, the nuclear magnetic data were obtained using CDCl ₃、CD₃OD、D₂ O, DMS-d6, etc., as solvents, based on tetramethylsilane (0.000 ppm) or on residual solvent (CDCl _3:7.26ppm;CD₃OD:3.31ppm;D₂ O:4.79ppm; d6-DMSO:2.50 ppm), and when peak shape diversity was indicated, the following abbreviations indicate different peak shapes: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad), dd (doublet), dt (doublet). If the coupling constant is given, it is in Hertz (Hz).

Intermediate synthesis

Preparation of A1

Step A

In a dry 2L three-necked flask, sodium hydride (0.8 g,19 mmol) was added to N, N-dimethylformamide (10 mL), the reaction system was heterogeneous gray, cooled to 0℃and a solution of Compound A1-1 (1 g,8 mmol) in N, N-dimethylformamide (200 mL) was added dropwise under nitrogen protection, the reaction was continued at 0℃for 0.5 hours, p-methoxychlorobenzyl chloride (2.75 g.17mmol,2.4 mL) was added, the temperature was slowly raised to 20℃and stirring was continued under nitrogen protection for 7.5 hours. The reaction solution was slowly added to 10mL of saturated ammonium chloride, extracted with methyl t-butyl ether (10 mL. Times.2), the combined organic phases were washed with 10mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure, and the obtained crude product was separated and purified by silica gel column chromatography to give Compound A1-2.LC-MS (ESI) m/z=365.45 [ M+H ] ⁺.

Step B

2,2.6,6-Tetramethylpiperidine (1.14 g,8.21 mmol) was added to anhydrous tetrahydrofuran (300 mL), cooled to-5 ℃, N-butyllithium (2.5M, 4 mL) was added dropwise, reacted at 5~0 ℃for 15min, cooled to-60℃and a solution of Compound A1-2 (0.98 g, 2.7 mmol) in tetrahydrofuran (3 mL), reacted at-60℃for 0.5 h, N-dimethylformamide (4 g,0.05 mol) was added rapidly, and the reaction mixture was stirred for 10min at-60 ℃. To the reaction solution, 20mL of saturated ammonium chloride was added, extraction was performed with methyl t-butyl ether (10 mL. Times.2), the combined organic phases were washed with 10mL of saturated brine, dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, the obtained crude product was slurried with the solvent for 0.5 hour, filtered, the cake was dried, the filtrate was concentrated, and then separated and purified by silica gel column chromatography, and the cake and column chromatography were combined to obtain the compound A1-3.LC-MS (ESI) m/z=393.5 [ m+h ] ⁺.

Step C

Compounds A1-3 (0.98 g.2.5 mmol) were added to 10mL of N, N-dimethylformamide, bromosuccinimide (0.45 g,2.5 mmol) was added and the reaction stirred at 20℃for an additional 20 minutes. The reaction mixture was added to 15mL of water, extracted with methyl t-butyl ether (8 ml×2), the combined organic phases were washed with saturated brine (5 ml××2), dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, and the crude product was slurried with the mixed solvent for 0.5 hours, filtered, and the cake was dried to give compounds A1 to 4.LC-MS (ESI) m/z=472.4 [ M+H ] ⁺.

Step D

Compounds A1-4 (0.97 g.2.04 mmol) were added to N, N-dimethylformamide (9.5 mL), and the reaction mixture was added with copper iodide (0.78 g,4.1 mmol) and methyl fluorosulfonyl difluoroacetate (2 g,10 mmol) under nitrogen, and the reaction mixture was warmed to 100℃and stirred for an additional 1 hour. The reaction solution was cooled, filtered through celite, the filtrate was added to 15mL of water, extracted with methyl t-butyl ether (7.5 mL. Times.2), the combined organic phases were washed with saturated brine (10 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure, and the obtained crude product was separated and purified by silica gel column chromatography to give compound A1-5.LC-MS (ESI) m/z=461.5 [ M+H ] ⁺.

Step E

Anhydrous tetrahydrofuran (50 mL) and sodium hydride (0.3 g,7.3 mmol) were added to a dry three-necked flask, cooled to 0 ℃, ethyl acetate (0.85 g,7.3mmol,0.8 mL) was added dropwise under nitrogen protection, the reaction solution was stirred for 0.5 hours at 0 ℃, n-butyllithium (2.5 m,3 mL) was added dropwise, the reaction solution was stirred for 0.5 hours under this condition, cooled to-60 ℃, a solution of compound A1-5 (1.15 g,2.5 mmol) in tetrahydrofuran (5 mL) was added dropwise, and the reaction solution was stirred for 0.5 hours at-60 ℃. To the reaction solution was added 25mL of a saturated ammonium chloride solution, extracted with 10mL of ethyl acetate, the organic phase was washed with 20mL of a saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure, and the obtained crude product was separated and purified by silica gel column chromatography to obtain the compound A1-6.LC-MS (ESI) m/z=531.6 [ M+H ] ⁺.

Step F

Compounds A1-6 (1 g,1.9 mmol) were added to dichloromethane (10 mL), ethyl 3-mercaptopropionate (0.3 g,2.3 mmol) and titanium tetrachloride were added and the reaction stirred at room temperature for 48 hours. Filtering, removing the organic solvent under reduced pressure, and separating and purifying the obtained crude product by silica gel column chromatography to obtain the compound A1-7.LC-MS (ESI) m/z=665.7 [ M+H ] ⁺

Step G

Compounds A1-7 (0.80 g,1.2 mmol) and sodium ethoxide (0.57 mg,1.9 mmol) were dissolved in THF10 mL and the reaction stirred at room temperature for 24 hours. Filtering and drying to obtain A1-8.LC-MS (ESI) m/z=619.7m+h ⁺.

Step H

To a suspension of A1-8 (3.3 g,5.3 mmol) in water (10 mL) was added S-methyl isothiourea sulfate (1.7 g,6.2 mmol) and potassium carbonate (1.6 g,11.7 mmol) at room temperature. The reaction mixture was stirred for 20 hours. The precipitate was collected, washed with water (twice) and isopropyl ether and dried to give A1-9; LC-MS (ESI): m/z=645.7 [ M+H ] ⁺.

Step I

Compounds A1-9 (1 g,1.6 mmol) were dissolved in dichloromethane (10 mL), N diisopropylethylamine (0.6 g,4.8 mmol) was added, cooled to 0-10℃and trifluoromethanesulfonic anhydride (0.7 g,2.4mmol, 400. Mu.L) was slowly added to the reaction solution, and reacted at this temperature for 15 minutes. The reaction solution was poured into a saturated aqueous ammonium chloride solution (8 mL), the solution was separated, the aqueous phase was extracted with methylene chloride (5 mL. Times.2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, the resulting crude product was slurried, filtered, and the cake was dried to give A1.LC-MS (ESI) m/z=777.8 [ M+H ] ⁺.

Preparation of B1

Compound B1 was obtained by a similar preparation method to intermediate A1 (starting material was changed to 6-bromo-4-methylpyridin-2-amine). LC/MS (ESI) m/z=760.8 [ M+H ] ⁺.

Example 1

1- ((3S) -4- (7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) prop-2-en-1-one (Compound 1)

Step A

Compound A1 (82 mg, 105.02. Mu. Mol) and 8-t-butoxycarbonyl-3, 8-diazabicyclo [3.2.1] octane (26.63 mg, 126.03. Mu. Mol) were dissolved in N, N-dimethylformamide (2 mL), diisopropylethylamine (40.72 mg, 315.07. Mu. Mol) was added, and the reaction solution was heated to 100℃and stirred for an additional 1 hour. Cooling, decompressing and removing the organic solvent, and separating and purifying the obtained crude product by a thin layer chromatography preparation plate to obtain the compound 1-1.LC/MS (ESI) m/z=828 [ M+H ] ⁺.

Step B

Compound 1-1 (71 mg, 84.96. Mu. Mol) was dissolved in methylene chloride (2 mL), and m-chloroperoxybenzoic acid (34.50 mg, 169.92. Mu. Mol) was added, and the reaction mixture was stirred at 20℃for 3 hours. The organic solvent is removed under reduced pressure, and the obtained crude product is separated and purified by thin layer chromatography preparation plates to obtain the compound 1-2.LC/MS (ESI) m/z=892 [ M+H ] ⁺.

Step C

The compound ((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methanol (12.09 mg, 75.94. Mu. Mol) was dissolved in anhydrous toluene (1 mL) under ice-water bath conditions, sodium t-butoxide (7.30 mg, 75.94. Mu. Mol) was added, the reaction mixture was stirred for 30 minutes, a toluene (1 mL) solution of the compound 1-2 (53 mg, 58.42. Mu. Mol) was added, and the reaction mixture was stirred under ice-water bath for 2 hours. The organic solvent was removed under reduced pressure, and the crude product was purified by thin layer chromatography to give compounds 1-3.LC/MS (ESI) m/z=971.1 [ M+H ] ⁺.

Step D

Compounds 1-3 (42 mg, 42.78. Mu. Mol) were dissolved in anhydrous dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the reaction was stirred at 20℃for an additional 2 hours. The solvent was removed under reduced pressure, and the crude product was purified by high performance liquid chromatography to give compounds 1-4.LC/MS (ESI) m/z=630.7 [ M+H ] ⁺.

Step E

Compounds 1-4 (0.46 g,0.73 mmol) and N, N-diisopropylethylamine (0.48 g,3.75 mmol) were dissolved in dichloromethane (20 mL), acryloyl chloride (59.73 mg,0.66 mmol) was added at-78℃and stirred at-78℃for 25 min. The reaction was quenched with water and extracted with dichloromethane. The organic layers are mixed together. The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was directly purified by reverse phase chromatography to give the compound 1.LC-MS(ESI)：m/z＝684.7[M+H]⁺.¹H-NMR(CD₃OD)δ6.73-6.87(m,1H),6.68(d,1H),6.25(dd,1H),5.46-5.58(m,1H),5.19-5.46(m,1H),4.90-5.05(m,2H),4.58-4.74(m,2H),3.94-4.43(m,8H),3.24-3.79(m,4H),2.89-3.13(m,2H),2.03-2.32(9H),1.31-1.42(m,3H).

Example 2

1- ((3S) -4- ((7S) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) prop-2-en-1-one (compound 1A)

1- ((3S) -4- ((7R) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) prop-2-en-1-one (compound 1B)

The compound 1 was separated and purified by high performance liquid chromatography to give 1A and 2A (column: CHIRALPAK IG-3:3 μm,0.46 cm. Times.5 cm; mobile phase: A (CO ₂) and B (EtOH, 0.1% isopropylamine), gradient: B% = 5 to-50%, 3min, flow rate: 3.4mL/min, wavelength: 220n, pressure: 1800, LC/MS (ESI): m/z=684.7 [ M+H ] ⁺.

Example 3

1- ((3S) -4- (7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) -2-fluoroprop-2-en-1-one (Compound 2)

Compound 2 was obtained by a similar preparation as step E in example 1. LC/MS (ESI) m/z=702.7 [ m+h ] ⁺.

Example 4

1- ((3S) -4- ((7S) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) -2-fluoroprop-2-en-1-one (2A)

1- ((3S) -4- ((7R) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) -2-fluoroprop-2-en-1-one (2B)

Compounds 3A and 3B were obtained by a similar preparation to the compounds 1A and 1B of example 2 (starting material was changed to compound 2). LC/MS (ESI) m/z=702.7 [ m+h ] ⁺.

Example 5

2- ((2S) -1-propenoyl-4- (7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) pyrazin-2-yl) acetonitrile (compound 3)

A compound was obtained by a preparation method similar to that of Compound 1 in example 1 3.LC/MS(ESI):m/z＝709.7[M+H]⁺.¹H-NMR(CD₃OD)δ6.73-6.87(m,1H),6.68(d,1H),6.25(dd,1H),5.46-5.58(m,1H),5.19-5.46(m,1H),4.90-5.05(m,2H),4.58-4.74(m,2H),3.94-4.43(m,8H),3.24-3.79(m,4H),2.89-3.13(m,2H),2.16-2.82(m,9H).

Example 6

2- ((2S) -1-propenoyl-4- ((7S) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) pyrazin-2-yl) acetonitrile (compound 3A)

2- ((2S) -1-propenoyl-4- ((7R) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) pyrazin-2-yl) acetonitrile (compound 3B)

Compounds 3A and 3B were obtained by a similar preparation method to the compounds 1A and 1B in example 2 (starting material was changed to compound 3). LC/MS (ESI) m/z=709.7 [ M+H ] ⁺.

Example 7

2- ((2S) -4- (7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (2-fluoroacryloyl) pyrazin-2-yl) acetonitrile (compound 4)

The compound was obtained in a similar manner to step E in example 1 4.LC/MS(ESI):m/z＝727.7[M+H]⁺.¹H-NMR(CD₃OD)δ6.72-6.84(m,1H),5.46-5.58(m,1H),5.19-5.46(m,2H),4.90-5.05(m,2H),4.58-4.74(m,2H),3.94-4.43(m,8H),3.24-3.79(m,4H),2.89-3.13(m,2H),2.18-2.82(m,9H).

Example 8

2- ((2S) -4- ((7S) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (2-fluoroacryloyl) pyrazin-2-yl) acetonitrile (compound 4A)

2- ((2S) -4- ((7R) -7- (3-amino-2-fluoro-5-methyl-6- (trifluoromethyl) phenyl) -2- (((2R, 7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (2-fluoroacryloyl) pyrazin-2-yl) acetonitrile (compound 4B)

Compounds 4A and 4B were obtained by a similar preparation to compounds 1A and 1B of example 2 (starting material was changed to compound 4). LC/MS (ESI) m/z=727.7 [ M+H ] ⁺.

Example 9

1- ((3S) -4- (7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) prop-2-en-1-one (Compound 5)

Compound 5 was obtained by a similar preparation method to compound 1 in example 1 (starting material was changed to B1). LC/MS (ESI) m/z=6677 [ M+H ] ⁺.

Example 10

1- ((3S) -4- ((7S) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) prop-2-en-1-one (compound 5A)

1- ((3S) -4- ((7R) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) prop-2-en-1-one (compound 5B)

Compounds 5A and 5B were obtained by a similar preparation to compounds 1A and 1B of example 2 (starting material was changed to compound 5). LC/MS (ESI) m/z=667.7 [ M+H ] ⁺.

Example 11

1- ((3S) -4- (7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) -2-fluoroprop-2-en-1-one (Compound 6)

Compound 6 was obtained by a similar procedure to step E of example 1. LC/MS (ESI) m/z=685.7 [ M+H ] ⁺.

Example 12

1- ((3S) -4- ((7S) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) -2-fluoroprop-2-en-1-one (compound 6A)

1- ((3S) -4- ((7R) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 AS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -3-methylpyrazin-1-yl) -2-fluoroprop-2-en-1-one (compound 6B)

Compounds 6A and 6B were obtained by a similar preparation to the compounds 1A and 1B of example 2 (starting material was changed to compound 6). LC/MS (ESI) m/z=685.7 [ M+H ] ⁺.

Example 13

2- ((2S) -4- (7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (propenoyl) pyrazin-2-yl) acetonitrile (compound 7)

Compound 7 was obtained by a similar preparation method to compound 1 in example 1 (starting material was changed to B1). LC/MS (ESI) m/z=692.7 [ M+H ] ⁺.

Example 14

2- ((2S) -4- ((7S) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7A (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (propenoyl) pyrazin-2-yl) acetonitrile (compound 7A)

2- ((2S) -4- ((7R) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (propenoyl) pyrazin-2-yl) acetonitrile (compound 7B)

Compounds 7A and 7B were obtained by a similar preparation to the compounds 1A and 1B of example 2 (starting material was changed to compound 7). LC/MS (ESI) m/z=692.7 [ M+H ] ⁺.

Example 15

2- ((2S) -4- (7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (2-fluoroacryloyl) pyrazin-2-yl) acetonitrile (compound 8)

Compound 8 was obtained by a similar procedure to step E of example 1. LC/MS (ESI) m/z=710.7 [ M+H ] ⁺.

Example 16

2- ((2S) -4- ((7S) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (2-fluoroacryloyl) pyrazin-2-yl) acetonitrile (compound 8A)

2- ((2S) -4- ((7R) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -2- (((2R, 7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -6, 6-dioxo-7, 8-dihydro-5H-thiopyran [4,3-d ] pyrimidin-4-yl) -1- (2-fluoroacryloyl) pyrazin-2-yl) acetonitrile (compound 8B)

Compounds 8A and 8B were obtained by a similar preparation to the compounds 1A and 1B of example 2 (starting material was changed to compound 8). LC/MS (ESI) m/z=710.7 [ M+H ] ⁺.

EXAMPLE 17 biological Activity test

The invention is further explained below in connection with test examples, but these implementations are not meant to limit the scope of the invention.

1. Tumor cell proliferation inhibition assay

1. Experimental method

Cell density was determined by Scepter automatic cell counter after resuspension of H358 (KRAS G12C mutant) cells by digestion centrifugation, cells were diluted to 44,000 cells per ml, and the cell solution after density adjustment was added to 96-well plates at 90 μl per well. After cells were cultured for 24 hours in a 37 ℃ 5% co ₂ incubator, cells with different concentrations of the test compound were added and incubated with the compound in the presence of 10% fetal bovine serum for 72 hours, cell growth inhibition was assessed using CELL TITER-Glo luminescent cell viability assay kit as detailed in manufacturer's instructions) to determine ATP content, briefly 30 microliters of CELL TITER-Glo reagent was added to each well, shaking plates for 10 minutes, cell lysis was induced, fluorescent signal was recorded with FluoroskanAscentFL (Thermo) assay, and the maximum signal value was obtained from cells treated with dimethyl sulfoxide for 72 hours. The minimum signal value was obtained from the medium alone (cell number zero), inhibition%o = (maximum signal value compound signal value)/(maximum signal value—minimum signal value x 100%, data were processed using GRAPHPADPRISM software IC ₅₀ values were calculated by sigmoidal dose response curve fitting where "a" represents IC ₅₀ +≤10 nM, "B" represents 10< IC ₅₀ +≤100 nM, "C" represents 100< IC ₅₀ +.1000 nM, and "D" represents 1000nM < IC ₅₀

2. Experimental results

The results of the calculation of 1C ₅₀ for each of the compounds in the above experiments are shown in Table 1 below

Table 1, inhibitory activity of the compounds against tumor cell proliferation IC ₅₀ (nm).

In vitro kinase activity shows that the designed compounds have strong inhibition activity on KRAS ^G12C mutant strains.

Although the invention has been described in detail hereinabove, those skilled in the art will appreciate that various modifications and changes can be made thereto without departing from the spirit and scope of the invention. The scope of the invention is not limited by the detailed description set forth above, but rather is to be attributed to the claims.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, selected from any one of the following:

2. The compound of any one of claim 1, or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, a solvate thereof, wherein the pharmaceutically acceptable salt is selected from the group consisting of potassium salt, sodium salt, magnesium salt, calcium salt, sulfate, hydrochloride, phosphate, sulfonate, and carbonate.

3. A pharmaceutical composition comprising a compound of any one of claim 1, or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, a solvate, and a pharmaceutically acceptable carrier thereof.

4. Use of a compound according to any one of claims 1, or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, a solvate thereof, for the preparation of a pharmaceutical composition for the treatment of a disease, disorder or condition associated with KRas G12C activity or expression level.

5. The use according to claim 4, wherein the disease, disorder or condition is selected from the group consisting of pancreatic cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, colon cancer, colorectal cancer, thyroid cancer, embryonal rhabdomyosarcoma, skin granulosa cell tumor, melanoma, liver cancer, rectal cancer, bladder cancer, throat cancer, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous carcinoma, cervical cancer, esophageal cancer, renal cancer, skin cancer, lymphoma, gastric cancer, acute myeloid leukemia, myelofibrosis, B-cell lymphoma, monocytic leukemia, splenomegaly, eosinophilic leukocytosis syndrome multiple solid tumors and hematological tumors such as bone marrow cancer.