CN111377918B - KRAS inhibitor compound - Google Patents

Abstract

本发明提供的上述KRAS G12C抑制剂化合物对KRAS突变有较好的抑制作用，可以用于预防和/或治疗KRAS G12C介导的疾病。The present invention provides a compound having the structure of formula II or a pharmaceutically acceptable salt, ester, isomer, solvate, prodrug or isotopic label thereof:

The above KRAS G12C inhibitor compounds provided by the present invention have a good inhibitory effect on KRAS mutation, and can be used for preventing and/or treating KRAS G12C-mediated diseases.

Description

A KRAS inhibitor compound

technical field

The present invention relates to a novel KRAS G12C inhibitor compound and the use of the inhibitor compound to prevent or treat KRAS G12C-mediated diseases.

Background technique

Kirsten rat sarcoma virus homolog (KRAS) mutation was first described in NSCLC gene in 1984. It is a membrane-bound protein that is located inside the cell membrane; it is also located in the EGFR signaling pathway, which is important for the occurrence and development of tumors. It is very important. Under normal circumstances, KRAS protein and GDP are inactive. When the extracellular growth and differentiation factor transmits the signal to KRAS protein, it enhances its binding activity with GTP, so that the protein and GTP are combined into an activated state, and the signal system is opened. The growth, proliferation, angiogenesis and other processes of tumor cells require intracellular proteins for signal transduction, and the KRAS gene is the determinant of the transduction protein. The KRAS mutant encodes an abnormal protein, which stimulates and promotes the growth and spread of malignant tumor cells; Signaling effects of upstream EGFR. KRAS mutation promotes cell proliferation, transformation and anti-apoptosis by activating its downstream RAS-RAF-MEK-MAPK and P13K-AKT-mTOR and other cell signaling pathways, thereby leading to tumorigenesis and development.

According to COSMIC statistics, the incidence of KRAS gene point mutation accounts for about 30% of all human tumors, including 90% of pancreatic cancer, 45% of colon cancer, and 35% of non-small cell lung cancer. 80% of KRAS mutations occur at codon 12, causing a single amino acid substitution, namely, glycine (G) to alanine (A), cysteine (C), aspartic acid (D), serine ( S), arginine (R), and valine (V), with glycine (G) replaced by cysteine (C) the most common. The KRAS G12C mutant protein has a large proportion (14%) in lung cancer, especially non-small cell lung cancer; in addition, it is also expressed in some colorectal cancer (4%) and pancreatic cancer (2%) patients.

Due to the high expression of KRAS G12C mutation in tumor patients, it can also make patients resistant to other targeted drugs, which has attracted more and more experts and scholars' attention. However, the development of drugs directly targeting KRAS G12C target inhibitors is challenged by the complexity of biochemistry, which is synonymous with "undruggable" targets in oncology.

New drug discovery is a fast-moving field, with advances in technology accelerating the discovery of drug candidates. Among these candidate drugs, not only their pharmacodynamics needs to be evaluated, but also the drug metabolism and kinetic properties are very important new drug screening indicators. An ideal drug needs to have a durable drug action time and good bioavailability. Every year, a large number of drug candidates are eliminated due to their poor pharmacokinetic parameters and metabolic profiles. Therefore, the metabolic characteristics and pharmacokinetic parameters of a candidate drug are important evaluation indicators for whether it can be turned into a drug, and good pharmacokinetic parameters and metabolic characteristics are necessary for a promising lead compound. Therefore, it is possible to provide KRAS G12C inhibitors with good pharmacokinetic characteristics to exert pharmacodynamic effects in vivo more effectively.

SUMMARY OF THE INVENTION

Invention to solve problem

In order to solve the above technical problems, the purpose of the present invention is to provide a novel KRAS G12C inhibitor and the use of the inhibitor for the treatment of KRAS G12C-mediated diseases, such as cancer.

solution to the problem

In order to solve the above-mentioned technical problems, the present invention provides the following technical solutions:

In one aspect, the present invention provides a compound having the structure of formula II or a pharmaceutically acceptable salt, ester, isomer, solvate, prodrug or isotopic label thereof:

in,

R ₁ ' is selected from unsubstituted or R ₇ '-substituted C _6-10 aryl and 5-10 membered heteroaryl;

R ₂ ' is selected from unsubstituted or R ₈ '-substituted C _6-10 aryl and 5-10 membered heteroaryl;

R ₃ ' and R ₄ ' are each independently selected from hydrogen, deuterium, C _1-6 alkyl, or R ₃ and R ₄ are connected to form unsubstituted or optionally 1-3 selected from deuterium, halogen, hydroxyl, C 3-7-membered cycloalkyl or 3-7-membered heterocycloalkyl substituted by the substituent of _1-6 alkyl, or R ₃ and R ₄ form =O, =S, =N-CN or =CH ₂ ;

_R5 ' and R6' are each independently selected from hydrogen, deuterium and halogen _;

Each R ₇ ' and R ₈ ' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxyl, amino, C _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl ) ₂ , C _3-6 cycloalkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl, -COOC _1-6 alkyl, the amino, alkyl, cycloalkane alkenyl, alkenyl and alkynyl are unsubstituted or substituted with 1-3 substituents selected from halogen, hydroxy, amino, acetyl or deuterium atoms;

A is a 4-9 membered heterocyclic group unsubstituted or substituted by R ₉ ', each R ₉ ' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxyl, amino, C _1-6 alkyl, C _{1 -6} alkoxy, the amino group, alkyl group is unsubstituted or substituted with a substituent selected from 1-3 halogen, cyano, hydroxyl, amino or deuterium atoms.

环中4位的N原子。In a preferred embodiment, the above _- mentioned R ₁ ' is selected from unsubstituted or substituted C _6-10 aryl and 5-10 membered heteroaryl, and said R ₇ ' is at C _6-10 aryl ortho-substitution of the atom attached to the N atom in the 5- to 10-membered heteroaryl group, the N atom is

N atom at position 4 in the ring.

In a preferred embodiment, R1 _' is selected from unsubstituted or _R7' -substituted _C6-10 aryl and 5-6 membered heteroaryl comprising 1- 3 heteroatoms or heteroatom groups selected from N, O, S(O) _m , where m is an integer of 0-2.

In a preferred embodiment, the C _6-10 aryl group is selected from phenyl, naphthyl, tetrahydronaphthyl and 2,3-indenyl, preferably, the C _6-10 aryl group is phenyl; the 5-10-membered heteroaryl group is selected from thienyl, pyridyl, pyridine oxynitride, pyrimidinyl, pyrazinyl, pyridazinyl, pyridone, pyrazinone, pyrimidinone , pyridazinone, pyrrolyl, pyrazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, imidazolyl, tetrazolyl, isothiazolyl, oxazole base, isoxazolyl, thiadiazolyl, oxadiazolyl, naphthyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, quinolinyl, isoquinoline olinyl and quinazolinyl, preferably, the 5-10-membered heteroaryl group is selected from pyridyl or pyrimidinyl.

In a preferred embodiment, each _R7 ' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, _C1-6 alkyl, _C3-6 cycloalkyl and _C1-6 haloalkyl, preferably, each _R7 ' is independently selected from hydrogen, deuterium, methyl, _CH2F , _CHF2 , _CF3 , ethyl, propyl, isopropyl, butyl, sec-butyl, isopropyl Butyl, tert-butyl, cyclopropane, cyclobutane, cyclopentane and cyclohexane.

_In a preferred embodiment, R2' is selected from unsubstituted or _R8' -substituted _C6-10 aryl and 5-6 membered heteroaryl comprising 1- 3 heteroatoms or heteroatom groups selected from N, O, S(O) _r , wherein r is an integer of 0-2.

In a preferred embodiment, R ₂ ' is selected from unsubstituted or R ₈ '-substituted C _6-10 aryl and 5-10 membered heteroaryl; the C _6-10 aryl is selected from phenyl , naphthyl, tetrahydronaphthyl and 2,3-indenyl; the 5-10-membered heteroaryl group is selected from thienyl, pyridyl, pyridine oxynitride, pyrimidinyl, pyrazinyl, pyridazine base, pyridinone, pyrazinone, pyrimidinone, pyridazinone, pyrrolyl, pyrazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, imidazolyl, tetrazolyl, isothiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, Benzofuranyl, quinolinyl, isoquinolinyl and quinazolinyl, preferably, R ₂ ' is selected from unsubstituted or R ₈ 'substituted phenyl, imidazolyl, pyrrolyl, pyridine oxynitride , pyridyl, pyridone, naphthyl, quinolinyl, isoquinolinyl and quinazolinyl.

In a preferred embodiment, each R ₈ ' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxyl, amino, C _1-6 alkyl, -NHC _1-6 alkyl, -N( C _1-6 alkyl) ₂ , C _1-6 alkoxy, said amino, alkyl, unsubstituted or substituted with 1-3 substituents selected from halogen, hydroxyl, amino, acetyl or deuterium atoms , preferably, each R ₈ ' is independently selected from hydrogen, deuterium, fluorine, chlorine, hydroxyl and amino.

In a preferred embodiment, R ₃ ' and R ₄ ' are each independently selected from hydrogen, deuterium, C _1-6 alkyl, or R ₃ and R ₄ are joined to form cyclopropyl, or R ₃ ' and R ₄ ' forms =O, =S or =N-CN, preferably, R ₃ ' and R ₄ ' form =O.

_In a preferred embodiment, _R5 ' and R6' are each independently selected from hydrogen, deuterium, fluorine and chlorine.

In a preferred embodiment, A is a 4-9 membered heterocyclyl group that is unsubstituted or substituted with _R9 ', respectively, and the atom to which the 4-9 membered heterocyclyl group is attached to the carbonyl group is N.

In a preferred embodiment, A is a 4-9-membered heterocyclic group unsubstituted or substituted by R ₉ ′, and the 4-9-membered heterocyclic group includes a monocyclic ring, a fused ring, a bridged ring, a spirocyclic ring .

In a preferred embodiment, A is a 6-7 membered heterocyclyl group that is unsubstituted or substituted with 1-3 R ₉ ', respectively, the 6-7 membered heterocyclyl group contains no double bonds or contains 1 or 2 double bonds, preferably, A is a 6-7-membered heterocycle that is unsubstituted or substituted by 1-2 R ₉ ' respectively, and the 6-7-membered heterocycle is selected from

In a preferred embodiment, each _R9 ' is independently selected from hydrogen, deuterium, methyl, ethyl, _-CH2OH , _-CH2CN and _-CH2F .

In a preferred embodiment, A is the following group:

In a preferred embodiment, R ₁ ' is selected from unsubstituted or R ₇ '-substituted C _6-10 aryl and 5-10 membered heteroaryl;

R ₂ ' is selected from unsubstituted or R ₈ '-substituted C _6-10 aryl and 5-10 membered heteroaryl;

R ₃ ' and R ₄ ' are each independently selected from hydrogen, deuterium, C _1-6 alkyl, or R ₃ and R ₄ are connected to form cyclopropyl, or R ₃ and R ₄ form =O;

_R5 ' and R6' are each independently selected from hydrogen, deuterium and halogen _;

each R ₇ ' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C _1-6 alkyl, C _3-6 cycloalkyl;

Each R ₈ ' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , C _1-6 alkoxy, said amino, alkyl, unsubstituted or substituted by 1-3 halogen, hydroxyl, amino, acetyl or deuterium atoms;

A is a 6-7 membered heterocyclic ring that is unsubstituted or substituted by 1-3 R ₉ ', respectively, and the atom connecting the 6-7 membered heterocyclic ring to the carbonyl group is N, and each R ₉ ' is independently selected from hydrogen , deuterium, methyl, ethyl, _-CH2OH and _-CH2F .

In a more preferred embodiment, R ₁ ' is selected from unsubstituted or R ₇ '-substituted C _6-10 aryl and 5-10 membered heteroaryl; the C _6-10 aryl is selected from benzene base, naphthyl, tetrahydronaphthyl and 2,3-indenyl; the 5-10-membered heteroaryl group is selected from thienyl, pyridyl, pyridine oxynitride, pyrimidinyl, pyrazinyl, pyridyl Azinyl, pyridinone, pyrazinone, pyrimidinone, pyridazinone, pyrrolyl, pyrazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl , imidazolyl, tetrazolyl, isothiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl , benzofuranyl, quinolinyl, isoquinolinyl and quinazolinyl;

R ₂ ' is selected from unsubstituted or R ₈ '-substituted C _6-10 aryl and 5-10 membered heteroaryl; the C _6-10 aryl is selected from phenyl, naphthyl, tetrahydronaphthyl and 2,3-indenyl; the 5-10-membered heteroaryl group is selected from thienyl, pyridyl, pyridine oxynitride, pyrimidinyl, pyrazinyl, pyridazinyl, pyridone, pyrazinone base, pyrimidinonyl, pyridazinone, pyrrolyl, pyrazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, imidazolyl, tetrazolyl, iso Thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, quinolinyl, isoquinolinyl and quinazolinyl;

R ₃ ' and R ₄ ' form =0;

_R5 ' and R6' are each independently selected from hydrogen, deuterium, chlorine and fluorine _;

Each _R7 ' is independently selected from hydrogen, deuterium, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, cyclopropane, cyclobutane, cyclopentane alkane and cyclohexane;

each _R8 ' is independently selected from hydrogen, deuterium, fluoro, chloro, hydroxy, and amino;

每一个R₉’独立地选自氢、氘、甲基、乙基、-CH₂CN、-CH₂OH和-CH₂F。A is a 6-7-membered heterocyclic ring that is unsubstituted or substituted by 1-2 R ₉ ' respectively, and the 6-7-membered heterocyclic group is selected from

Each _R9 ' is independently selected from hydrogen, deuterium, methyl, ethyl, _-CH2CN , _-CH2OH , and _-CH2F .

The present invention also provides a compound or a pharmaceutically acceptable salt, ester, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotopic label or prodrug thereof, wherein the The compound is any of the following:

The present invention also provides a pharmaceutical composition comprising the above compound or a pharmaceutically acceptable salt, ester, isomer, solvate, hydrate, prodrug or isotopic label thereof.

In addition, the present invention also provides the above-mentioned compounds or their pharmaceutically acceptable salts, esters, hydrates, solvates, stereoisomers, tautomers, cis-trans isomers, isotopic labels or prodrugs, Or the application of the above pharmaceutical composition in the preparation of a medicine for preventing and/or treating KRAS G12C-mediated diseases, preferably, the diseases include lung cancer, pancreatic cancer, pancreatic ductal cancer, colon cancer, rectal cancer, appendix cancer, Esophageal squamous cell carcinoma, head and neck squamous cell carcinoma, breast cancer and other solid tumors.

In order to describe the content of the present invention more clearly, the terms involved are now defined as follows:

In the present invention, the term "C _1-6 alkyl" alone or in combination means a saturated straight or branched alkyl group containing 1-6, especially 1-4 carbon atoms, including methyl, ethyl propyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3- Methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, n-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3 -Methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butanyl base, 3,3,-dimethyl-2-butyl, etc. Preferably, "C _1-6 alkyl" is any one of methyl, ethyl, n-propyl, isopropyl, tert-butyl. Similarly, the term "C _1-3 alkyl" alone or in combination means a saturated straight or branched chain alkyl group containing 1-3 carbon atoms, including methyl, ethyl, propyl, isopropyl, etc. .

The term "3-7 membered cycloalkyl" alone or in combination denotes a cycloalkyl group having 3 to 7, especially 3 to 6 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl , cycloheptyl, etc. Particular " _{C3-7cycloalkyl} " are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

The term "amino" alone or in combination denotes a primary ( _-NH2 ), secondary (-NH-) or tertiary amino group

The term "C _1-6 alkoxy" alone or in combination means the group C _1-6 alkyl-O-, wherein "C _1-6 alkyl" means as defined above, which includes (but is not limited to) Methoxy (-OCH ₃ ), ethoxy (-OCH ₂ CH ₃ ), n-propoxy (-OCH ₂ CH ₂ CH ₃ ), isopropoxy (-OCH(CH ₃ ) ₂ ), n-butyl Oxy (-OCH ₂ CH ₂ CH ₂ CH ₃ ), sec-butoxy (-OCH(CH ₃ )CH ₂ CH ₃ ), isobutoxy (-OCH ₂ CH(CH ₃ ) ₂ ), tert-butoxy group (-OC(CH ₃ ) ₃ ), n-pentyloxy (-OCH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), neopentyloxy (-OCH ₂ C(CH ₃ ) ₃ ) and the like.

The term "halogen" alone or in combination means fluorine, chlorine, bromine or iodine. Particular is fluorine, chlorine or bromine.

The term "heterocycloalkyl", also known as "heterocyclyl", refers to a saturated or partially unsaturated (containing 1 or 2 double bond) non-aromatic ring consisting of carbon atoms and heteroatoms such as nitrogen, oxygen or sulfur The cyclic group can be a monocyclic, bicyclic bridged ring or a spirocyclic group. In the present invention, the number of carbon atoms in the heterocycloalkyl group is 2-11, and the number of heteroatoms is preferably 1, 2, 3 or 4, the nitrogen, carbon or sulfur atom in the heterocycloalkyl group can be optionally oxidized. The hydrogen atoms on "heterocycloalkyl" are independently optionally substituted with one or more substituents described herein. A "heterocycloalkyl" can be linked to the parent molecule through any ring atom in the ring.

The term "4-9 membered heterocyclyl" refers to a monocyclic group containing 4-9, especially 6-7 carbon atoms and a heteroatom or heteroatom group without double bonds or with 1 or 2 double bonds Rings, fused rings, bridged rings, spiro rings, the heteroatom or heteroatom group is selected from N, O, S(O) _m (wherein m is an integer from 0 to 2); for example no double bond or containing 1 azetidine, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, tetrahydro Hydropyranyl, 1,1-dioxothiomorpholinyl, bicyclo[4.1.0]heptyl, etc.

The term "aryl" refers to any stable 6-10 membered monocyclic or bicyclic aromatic group, including phenyl, naphthyl, tetrahydronaphthyl, 2,3-indenyl, or biphenyl, and the like. The hydrogen atoms on the "aryl group" are independently optionally substituted with one or more substituents described herein.

)、吡啶酮基、吡嗪酮基、嘧啶酮基、哒嗪酮基、吡咯基、吡唑基、噻唑基、1,2,3-三唑基、1,2,4-三唑基、咪唑基、四氮唑基、异噻唑基、噁唑基、异噁唑基、噻二唑基、噁二唑基、苯并噻吩基、吲哚基、苯并咪唑基、苯并噻唑基、苯并呋喃基、喹啉基、异喹啉基、喹唑啉基等。“杂芳基”上的氢原子独立任选地被一个或多个本发明所描述的取代基所取代。The term "heteroaryl" refers to an aromatic ring group formed by replacing a carbon atom on the ring with at least one heteroatom or heteroatom group selected from N, O, S(O) _m (where m is an integer from 0 to 2). The aromatic heterocyclic group may be a 5-7 membered monocyclic or 7-12 bicyclic group. In the present invention, the number of heteroatoms in the heteroaryl group is preferably 1, 2, 3 or 4, such as thienyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyridine oxynitride (ie

), pyridone, pyrazinone, pyrimidinone, pyridazinone, pyrrolyl, pyrazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, imidazolyl, tetrazolyl, isothiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, Benzofuranyl, quinolinyl, isoquinolinyl, quinazolinyl and the like. The hydrogen atoms on a "heteroaryl group" are independently optionally substituted with one or more substituents described herein.

The term "5-10 membered heteroaryl" refers to a heteroaromatic ring having 5-10 carbon atoms and a heteroatom or heteroatom group, wherein the heteroaromatic ring is as defined above. Similarly, the term "5-6 membered heteroaryl" refers to a heteroaromatic ring having 5-6 carbon atoms and a heteroatom or heteroatom group, wherein heteroaromatic ring is as defined above.

The term " _C6-10 aryl" means an aryl group having 6-10 carbon atoms, wherein aryl means as defined above.

The term "cyano" alone or in combination refers to the group -CN.

The term "hydroxy" alone or in combination refers to the group -OH.

The term "isomer" includes all isomeric forms including enantiomers, diastereomers, tautomers and geometric isomers (including cis-trans isomers). Therefore, the single stereochemical isomers of the compounds contemplated in the present invention or their enantiomers, diastereomers, tautomers or geometric isomers (or cis-trans isomers) Mixtures are within the scope of the present invention.

The term "pharmaceutically acceptable salts" means that the compounds of the present invention exist in the form of their pharmaceutically acceptable salts, including acid addition salts and base addition salts. Pharmaceutically acceptable salts are described in SM Berge in J. Pharmaceutical Sciences (Vol. 66: pp. 1-19, 1977). In the present invention, pharmaceutically acceptable non-toxic acid addition salts refer to salts formed by the compounds of the present invention with organic or inorganic acids, including but not limited to hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodide Acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, methanesulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid, malic acid, etc. Pharmaceutically acceptable non-toxic base addition salts refer to salts formed by the compounds of the present invention with organic or inorganic bases, including but not limited to alkali metal salts such as lithium, sodium or potassium salts; alkaline earth metal salts such as calcium Or magnesium salts; organic base salts, such as _ammonium salts or N ⁺ (C _1-6 alkyl) salts formed by organic bases containing N groups, preferably lithium hydroxide, sodium hydroxide, potassium hydroxide, Sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, calcium carbonate, ammonia water, triethylamine, tetrabutylammonium hydroxide, etc.

The term "solvate" refers to an association of one or more solvent molecules with a compound of the present invention. Solvates forming solvents include, but are not limited to, water, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, dimethylsulfoxide, and the like. "Pharmaceutically acceptable salts" can be synthesized by conventional chemical methods.

The term "ester" is used to refer to organic esters, including monoesters, diesters, triesters, and more generally polyesters.

The term "hydrate" refers to the association of water with the compounds of the present invention.

The term "prodrug" refers to a chemical derivative of a compound of the present invention which is converted into a compound represented by general formula I, II or III by chemical reaction in vivo.

The term "isotopic derivatives" refers to isotopic derivatives obtained by replacing the hydrogen atoms in the general formula I with 1-6 deuterium atoms (D), and the carbon atoms in the general formula (I) with 1-3 carbon 14 atoms ( ¹⁴ C) substituted isotopic derivatives.

The terms involved in the present invention are defined above, and those skilled in the art can also understand the above terms in combination with the prior art. The following further describes the terms based on the content of the present invention and the definitions of the terms.

The following examples may further describe the present invention, however, these examples should not be construed as limiting the scope of the present invention.

Detailed ways

Example 1SZ-014096

Step 1: Synthesis of compound 014004A2

Compound 2,6-dichloro-5-fluoronicotinic acid 014004A1 (16.0 g, 76.4 mmol) was dissolved in dichloromethane (200 mL), and oxalyl chloride (12.1 g, 95.2 mmol) was added dropwise in an ice-water bath, followed by a dropwise addition of catalytic The amount of DMF (0.2mL) was reacted at room temperature (20°C) overnight and then concentrated under reduced pressure. The obtained residue was dissolved in 200mL of 1,4-dioxane and cooled to zero degrees. Aqueous ammonia solution (28.0-30%, 14.4mL, 114.2 mmol) was slowly added dropwise to the reaction phase, stirred at zero for 30 minutes, concentrated under reduced pressure to remove the solvent, and the obtained crude white solid was purified by silica gel column chromatography (dichloromethane/methanol=100:1) to obtain a white solid compound 014004A2 (9.0 g, yield 56.2%). LCMS (M+H) ⁺ m/z calculated 209.0, found 209.0.

Step 2: Synthesis of compound 014004A4

To a suspension of compound 2-bromo-4-methyl-3-aminopyridine 014004A3 (5.4 g, 28.9 mmol) in tetrahydrofuran (60 mL) was added Pd(dppf)Cl ₂ (1.02 g, 1.4 mmol), the reaction mixture was Isopropylmagnesium chloride (2M in tetrahydrofuran, 22 mL, 43.4 mmol) was added at zero degree under nitrogen protection, the addition was completed, and the reaction was stirred at 60 degree overnight. The reaction was stopped and cooled to room temperature, the reaction solution was quenched with a mixture of 100 mL of water and 200 mL of 1N sodium hydroxide solution, extracted with ethyl acetate (200 mL*2), the combined organic phases were concentrated under reduced pressure, and the concentrated residue was passed through a silica gel column layer Analysis (petroleum ether/ethyl acetate=1:1) was purified to obtain compound 014004A4 (3.22 g, yield 73.1%) as a yellow colloid. LCMS (M+H) ⁺ m/z calcd 151.1, found 151.1. ¹ H NMR (DMSO-d ₆ , 400 MHz): δ 8.24 (d, J=8.0 Hz, 1H), 8.11 (br s, 1H), 7.95 (br s, 1H), 2.17 (s, 3H).

Step 3: Synthesis of compound 014004A5

Compound 014004A2 (5.0 g, 23.9 mmol) was dissolved in tetrahydrofuran (20 mL), and oxalyl chloride (3.66 g, 28.8 mmol) was slowly added under an ice-water bath. The reaction mixture was stirred at 75 degrees for 1 hour. Stop heating and lower the temperature to zero, slowly add compound 014004A4 (3.6 g, 23.9 mmol) in tetrahydrofuran (10 mL) dropwise, after the drop is complete, the reaction continues to stir at zero for 1 hour, followed by a 1:1 mixed solution of brine and ammonium chloride Quenched, extracted with ethyl acetate (200 mL*3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue 014004A5 (9.1 g, crude product), which was used in the next step without purification. LCMS (M+H) ⁺ m/z calcd 385.1, found 385.1.

Step 4: Synthesis of compound 014004A6

The crude compound 014004A5 (9.1 g, 23.8 mmol) was dissolved in tetrahydrofuran (40 mL), KHMDS (1 M in THF, 50.2 mL, 50.2 mmol) was slowly added dropwise under an ice bath, the ice bath was removed, and the reaction solution was kept at room temperature (20° C. ) stirred for 1 hour, the reaction phase was quenched with saturated ammonium chloride solution, extracted with ethyl acetate (100 mL*3), the organic phases were combined and dried with anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure and the residue obtained was passed through silica gel column chromatography (petroleum Ether/ethyl acetate/dichloromethane/methanol=100:10:10::1) was purified to obtain compound 014004A6 (3.2 g, yield 38.2%) as a pale yellow solid. LCMS (M+H) ⁺ m/z calcd 349.1, found 349.1. ¹ H NMR (DMSO-d ₆ , 400MHz): δ 12.27 (br s, 1H), 8.53-8.49 (m, 2H), 7.29 (d, J=4.8Hz, 1H), 2.87 (q, J=6.6 Hz, 1H), 2.05-1.99 (m, 3H), 1.09 (d, J=6.6Hz, 3H), 1.01 (d, J=6.6Hz, 3H).

Step 5: Synthesis of compound 014004A7

Compound 014004A6 (2.1 g, 6.0 mmol) and N,N-diisopropylethylamine (1.16 g, 9.0 mmol) were dissolved in acetonitrile (10 mL), phosphorus oxychloride (0.72 mL, 7.8 mmol) in an ice bath It was slowly added dropwise at 80° C. for 1 hour, cooled to room temperature and spin-dried to obtain a brown oily residue 014004A7 (2.2 g, crude product), which was directly used in the next step without purification. LCMS (M+H) ⁺ m/z calculated 367.0, found 367.0.

Step 6: 014004A8 Synthesis

The crude compound 014004A7 (2.2 g, 6.0 mmol) was dissolved in acetonitrile (10 mL), and N,N-diisopropylethylamine (2.3 g, 18.0 mmol) and (S)-4-N-tert. Butoxycarbonyl-2-methylpiperazine (1.44 g, 7.2 mmol), the ice bath was removed, and the reaction solution was stirred at room temperature (20 °C) for 1 hour, followed by the addition of ice-saturated sodium bicarbonate solution (100 mL) and ethyl acetate Ester (150mL), continue to stir for 5 minutes, separate the two phases, the aqueous phase continues to be extracted with ethyl acetate (100mL*2), the organic phases are combined and dried over anhydrous sodium sulfate, the filtrate is concentrated under reduced pressure and the residue obtained is passed through silica gel column chromatography (Petroleum ether/ethyl acetate/dichloromethane/methanol=80:10:10::1) Purification gave a pale yellow solid compound 014004A8 (1.8 g, 56.3% yield). LCMS (M+H) ⁺ m/z calcd 531.2, found 531.2. ¹ H NMR (CDCl ₃ , 400 MHz): δ 8.56 (d, J=4.8 Hz, 1H), 7.78 (d, J=7.2 Hz, 1H), 7.13 (d, J=4.8 Hz, 1H), 4.79 ( br s, 1H), 4.10 (br s, 3H), 3.64 (br s, 1H), 3.21 (br s, 2H), 2.64-2.58 (m, 1H), 2.04-2.02 (m, 3H), 1.54 ( s, 9H), 1.51-1.42 (m, 3H), 1.28-1.21 (m, 3H), 1.15-1.07 (m, 3H).

Step 7: 014004A9 Synthesis

Compound 014004A8 (1.7 g, 3.21 mmol), 2-fluoro-6-hydroxyphenylboronic acid (605.6 mg, 3.91 mmol), potassium acetate (1.62 g, 16.5 mmol) and Pd(dppf)Cl2 (119 mg, 0.17 mmol) were dissolved In 1,4-dioxane (25 mL), after nitrogen replacement for several times, the reaction solution was stirred at 90° C. for 1 minute, then 2 drops of water were added, continued to stir at 90° C. for 1 hour, cooled to room temperature, and water (25 mL) was added. ) and brine (25ml) were added to the reaction phase, extracted with ethyl acetate (200mL*2), the organic phases were combined and dried over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure and the residue obtained was subjected to silica gel column chromatography (petroleum ether/acetic acid) Ethyl ester/dichloromethane/methanol=60:10:10::1) was purified to give yellow solid compound 014004A9 (1.6 g, 82.5% yield).

Subsequent separation by chiral preparative (CO ₂ :EtOH=60/40) by supercritical chromatography gave yellow solid compounds 014004A9A (1st eluting isomer) (700 mg) and 014004A9B (2nd eluting isomer) (650 mg) ). LCMS (M+H) ⁺ m/z calculated 607.3, found 607.3.

*250mmL*5um；Method：CO₂:EtOH＝60/40Chiral preparation conditions: Column: IC

*250mmL*5um; Method: CO ₂ :EtOH＝60/40

Flow: 50 g/min; λ: 214 nm; Rt(014004A9A): 3.811, Rt(014004A9B): 6.588.

Step 8: Synthesis of 014004A10A and 014004A10B

Compound 014004A9A (700 mg, 1.15 mmol) was dissolved in dichloromethane (6 mL), trifluoroacetic acid (2.8 mL, 35 mmol) was added under an ice bath to remove the ice bath, and the reaction solution was stirred at room temperature (20° C.) for 1 hour, and then rotated. Dry the reaction solution, add ice-saturated sodium bicarbonate solution (10 mL) to the brown oily residue, extract with dichloromethane (100 mL*2), and wash with water (50 mL*2). The organic phase was dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the residue 014004A10A (550 mg, crude product, 94.2% yield), which was used directly in the next step without purification. LCMS (M+H) ⁺ m/z calcd 507.2, found 507.2.

Compound 014004A9B (650 mg, 1.07 mmol) was dissolved in dichloromethane (6 mL), trifluoroacetic acid (2.6 mL, 32.5 mmol) was added under an ice bath to remove the ice bath, and the reaction solution was stirred at room temperature (20° C.) for 1 hour, The reaction solution was spin-dried, ice-saturated sodium bicarbonate solution (10 mL) was added to the obtained brown oily residue, extracted with dichloromethane (100 mL*2), and washed with water (50 mL*2). The organic phase was dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the residue 014004A10B (510 mg, crude product, 94.1% yield), which was used in the next step without purification. LCMS (M+H) ⁺ m/z calcd 507.2, found 507.2.

Step 9: 014096A1 Synthesis

2-Methylfuran (2.0 g, 24.4 mmol) was dissolved in tert-butanol-water (5:1, v/v, 120 mL), and sodium dihydrogen phosphate (5.7 g, 36.6 mmol) sodium chlorite was added with stirring (6.6 g, 73.2 mmol), the reaction phase was stirred at room temperature (25 degrees) for two hours or until the yellow color disappeared. The reaction solution was removed by rotary evaporation, the obtained residue was extracted with chloroform (300ml), washed with brine (2*200ml), the organic phase was dried with magnesium sulfate, filtered and the solvent was rotated to dryness, the obtained yellow oily residue 014096A1 was directly used in the next step without purification. one-step reaction. (300 mg, crude yield 10.8%). LCMS (M+H) ⁺ m/z calcd 115.0, found 115.1.

Step 10: 014096A2 Synthesis

Compound 014096A1 (300mg, 2.63mmol) was dissolved in tetrahydrofuran-acetone-water (5:4:1, 40ml), freshly distilled pyridine (1mol%, 200uL) was added, and the reaction phase was stirred at room temperature (25 degrees) for two hours , the oily crude product obtained by concentrating the reaction solution was purified by silica gel column chromatography (ethyl acetate/petroleum ether=1:2) to obtain a yellow oily crude product compound 014096A2 (80 mg, yield 26.7%). LCMS (M+H) ⁺ m/z calcd 115.0, found 115.1.

Step 11: Synthesis of SZ-014096

Compound 014009A10A (100 mg, 0.2 mmol), 014096A2 (22 mg, 0.2 mmol), HATU (112 mg, 0.296 mmol), dissolved in N,N-dimethylformamide (2.0 mL), added N,N-diiso Propylethylamine (51 mg, 0.4 mmol). The reaction phase was stirred at room temperature (25 degrees) for two hours. N,N-dimethylformamide was removed by concentration under reduced pressure, and the concentrated residue was purified by preparative high performance liquid chromatography to obtain a white solid compound SZ-014096 (17.0 mg, yield 14.3%).

Liquid mass spectrometry [mobile phase: gradient from 70% water (with 0.02% ammonium acetate) and 30% acetonitrile to 50% water (with 0.02% ammonium acetate) and The 50% acetonitrile elution was maintained for 6.5 minutes. Column: waters XBridge C18 3.5um, 50*4.6mm] purity 96.76%, Rt=2.889min; LCMS (M+H) ⁺ m/z calculated 603.3, found 603.3. ¹ H NMR (DMSO-d ₆ , 400 MHz): δ 10.20 (br s, 1H), 8.39 (d, J=4.8 Hz, 1H), 8.27 (dd, J ₁ =21.6 Hz, J ₂ =9.2 Hz, 1H), 7.45 (q, J=15.6Hz, 1H), 7.25 (dd, J1 ₌ 15.2Hz, J2=8.0Hz, 1H), 7.19 (d, J=4.8Hz, 1H), _6.81-6.66 ( m, 3H), 4.98-4.91 (m, 1H), 4.36 (q, J=14.0Hz, 2H), 4.21-4.04 (m, 1H), 3.77-3.52 (m, 2H), 3.23-3.20 (m, 1H), 2.74-2.65(m, 1H), 2.43(s, 3H), 1.90(d, J=2.4Hz, 3H), 1.36(d, J=6.4Hz, 3H), 1.07(d, J=6.4 Hz, 3H), 0.93 (d, d, J=6.8 Hz, 3H).

Example 2SZ-014096B

The first step: SZ-014096B synthesis

Compounds 014004A10B (100 mg, 0.2 mmol), 014096A2 (22 mg, 0.2 mmol), HATU (112 mg, 0.296 mmol) were dissolved in N,N-dimethylformamide (2.0 mL), to which was added N,N-bismuth Isopropylethylamine (51 mg, 0.4 mmol). The reaction phase was stirred at room temperature (25 degrees) for two hours. N,N-dimethylformamide was removed by concentration under reduced pressure, and the concentrated residue was purified by preparative high performance liquid chromatography to obtain a white solid compound SZ-014096B (23.0 mg, yield 19.3%). Liquid mass spectrometry [mobile phase: gradient from 80% water (with 0.02% ammonium acetate) and 20% acetonitrile to 30% water (with 0.02% ammonium acetate) and The 70% acetonitrile elution was maintained for 6.5 minutes. Column: waters XBridge C18 3.5um, 50x4.6mm] purity 99.14%, Rt=3.361 min; LCMS (M+H) ⁺ m/z calculated 603.3, found 603.2. ¹ H NMR (DMSO-d ₆ , 400 MHz): δ 10.18 (br s, 1H), 8.39 (d, J=4.8 Hz, 1H), 8.30 (dd, J ₁ =19.6 Hz, J ₂ =9.2 Hz, 1H), 7.46 (q, J=15.6Hz, 1H), 7.27 (dd, J1 ₌ 15.2Hz, J2=8.0Hz, 1H), 7.19 (d, J=4.8Hz, 1H), _6.82-6.66 ( m, 3H), 5.01-4.93 (m, 1H), 4.40-4.29 (m, 2H), 4.21-4.05 (m, 1H), 3.80-3.51 (m, 2H), 3.22-3.16 (m, 1H), 2.73-2.67(m, 1H), 2.39(s, 3H), 1.90(d, J=4.0Hz, 3H), 1.34(d, J=6.0Hz, 3H), 1.07(d, J=6.8Hz, 3H) ), 0.93 (d, J=6.0 Hz, 3H).

Example 3SZ-014017A/B

The first step: 014086A1 synthesis

The compound 4,6-dichloro-5-aminopyrimidine (10.0 g, 60.98 mmol) was dissolved in 200 mL of tetrahydrofuran, and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride was added (8.9 g, 12.2 mmol), isopropylmagnesium chloride (1N, 183 mL, 366 mmol) was added dropwise at zero degrees, and heated to 70 degrees overnight under nitrogen protection. The reaction solution was cooled to room temperature, quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=3:1 ) to give a yellow solid 014086A1 (3.0 g, 27% yield). LCMS (M+H) ⁺ m/z calcd 180.1, found 180.1. ¹ H NMR (DMSO-d ₆ , 400 MHz): δ 8.34 (s, 1H), 5.06 (s, 2H), 3.29-3.20 (m, 2H), 1.17 (d, J=8.8 Hz, 12H).

The second step: 014086A2 synthesis

Compound 2,6-dichloro-3-carboxamide-5-fluoropyridine (3.0 g, 14.4 mmol) was dissolved in 50 mL of tetrahydrofuran, oxalyl chloride (2.2 g, 17.28 mmol) was added at room temperature, and the reaction was heated under reflux for 1 hour. . The reaction solution was cooled to zero, compound 014086A1 (2.6 g, 14.4 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was adjusted to neutral pH with saturated sodium bicarbonate solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate= 5:1) to give a yellow solid 014086A2 (5.6 g, 90% yield). LCMS(M+H) ⁺ m/z calculated 414.2, found 414.2.

The third step: 014086A3 synthesis

Compound 014086A2 (5.2 g, 12.56 mmol) was dissolved in 50 mL of anhydrous tetrahydrofuran, KHMDS (1N, 27.63 mL) was added dropwise under ice bath, and the reaction was stirred at room temperature for 1 hour. The reaction solution was poured into water, adjusted to neutrality with 2N HCl, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10: 1) A pale yellow solid 014086A3 (4.3 g, yield 89%) was obtained. LCMS (M+H) ⁺ m/z calcd 378.2, found 378.2. ¹ H NMR (DMSO-d ₆ , 400MHz): δ 12.36 (s, 1H), 9.22 (s, 1H), 8.55 (d, J=10.0 Hz, 1H), 3.00-2.96 (m, 2H), 1.12 (d, J=8.8 Hz, 6H), 1.02 (d, J=8.8 Hz, 6H).

Step 4: 014086A4 Synthesis

Compound 014086A3 (800 mg, 2.02 mmol) was dissolved in 5 mL of anhydrous acetonitrile, POCl ₃ (487 mg, 3.18 mmol) and DIPEA (546 mg, 4.24 mmol) were added under ice bath, and the reaction was heated to 80 degrees overnight. The reaction solution was spin-dried to obtain brown oil 014086A4 (800 mg, yield 100%), and the crude product was directly used in the next step.

Step 5: 014086A5 Synthesis

The crude compound 014086A4 (800 mg, 2.02 mmol) was dissolved in 10 mL of acetonitrile, and (S)-4-N-tert-butoxycarbonyl-2-methylpiperazine (404 mg, 2.02 mmol) and DIPEA (781.74 mg, 6.06 mg) were added. mmol) and stirred at room temperature for 1 hour. The reaction solution was spin-dried, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=3:1) to obtain a pale yellow solid 014086A5 (600 mg, yield 54.5%). LCMS (M+H) ⁺ m/z calculated 560.2, found 560.2. ¹ H NMR (DMSO-d ₆ , 400MHz): δ 9.15 (s, 1H), 8.40 (d, J=8.4Hz, 1H), 4.93-4.91 (m, 1H), 4.28-4.27 (m, 1H) , 4.28-3.99(m, 1H), 4.06-3.97(m, H), 3.77-3.71(m, 1H), 3.26-3.17(m, 1H), 2.77-2.67(m, 2H), 1.56(s, 10H), 1.44-1.42 (m, 3H), 1.44-1.05 (m, 12H).

Step 6: 014086A6 Synthesis

Compound 014086A5 (560 mg, 1 mmol), 2-fluoro-6-hydroxyphenylboronic acid (1.24 g, 8 mmol) was dissolved in 10 mL of 1,4-dioxane, potassium acetate (490 mg, 5 mmol) and [1, 1'-bis(diphenylphosphino)ferrocene]dichloride palladium (146 mg, 0.2 mmol), microwave reaction at 120 degrees for 90 minutes. After cooling, the reaction solution was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=3:1) to obtain light yellow solid 014086A6 (300 mg, yield 47%). LCMS (M+H) ⁺ m/z calculated 636.3, found 636.3.

Step 7: 014086A7 Synthesis

Compound 014086A6 (230 mg, 0.36 mmol) was dissolved in 2 mL of dichloromethane, 2 mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was selected to dryness to obtain 230 mg of trifluoroacetate as a brown oil, which was directly used in the next step. LCMS (M+H) ⁺ m/z calculated 536.3, found 536.3.

The eighth step: SZ-014017A/B synthesis

Compound 014086A7 (50.0 mg, 0.09 mmol), 014096A2 (10.2 mg, 0.09 mmol), HATU (53.0 mg, 0.14 mmol) were dissolved in N,N-dimethylformamide (2.0 mL), and N,N was added - Diisopropylethylamine (35.0 mg, 0.27 mmol). The reaction phase was stirred at room temperature (25 degrees) for two hours. N,N-dimethylformamide was removed by concentration under reduced pressure, and the concentrated residue was purified by preparative high performance liquid chromatography to obtain white solid compound SZ-014017A/B (7.5 mg, yield 12.7%).

Liquid mass spectrometry [mobile phase: gradient from 70% water (with 0.02% ammonium acetate) and 30% acetonitrile to 40% water (with 0.02% ammonium acetate) and The 60% acetonitrile elution was maintained for 6.5 minutes. Column: waters XBridge C18 3.5um, 50*4.6mm] purity 99.14%, Rt=3.129min; LCMS (M+H) ⁺ m/z calculated 632.2, found 632.2. ¹ H NMR (DMSO-d ₆ , 400 MHz): δ 10.20 (br s, 1H), 9.05 (s, 1H), 8.32 (dd, J ₁ =22.0 Hz, J ₂ =9.2 Hz, 1H), 7.46 ( q, J=15.6Hz, 1H), 7.27 (dd, J1 ₌ 15.2Hz, J2=8.4Hz, 1H), _6.82-6.66 (m, 3H), 5.00-4.97 (m, 1H), 4.39-4.29 (m, 2H), 4.22-4.06 (m, 1H), 3.82-3.54 (m, 2H), 3.29-3.23 (m, 1H), 2.73-2.66 (m, 2H), 2.39 (s, 3H), 1.36 (d, J=6.8 Hz, 3H), 1.08 (d, J=6.8 Hz, 6H), 0.932 (d, J=6.4 Hz, 6H).

Biological Activity Experiment

Detection of KRAS G12C protein binding rate by LC-MS

Test compounds were prepared as 10 mM stock solutions in DMSO. KRAS G12C protein was diluted to 103 uM in buffer (20 mM Hepes, pH 7.5, 50 mM NaCl, 0.5 mM MgCl ₂ ) and an equal volume was added to GDP buffer (20 mM Hepes, pH 7.5, 50 mM NaCl, 0.5 mM MgCl ₂ , 10 mM EDTA) , 2mM DTT, GDP) was prepared as KRASG12C protein loaded with GDP.

A dilution solution (12.5 mM Hepes, pH 7.5, 75 mM NaCl, 10 mM MgCl ₂ ) was added to the GDP-loaded KRASG12C protein to dilute to 20 uM. The reaction system was prepared as follows: GDP-KRAS-4B-G12C (20 uM, 5 μL), test compound (10% DMSO, 5 μL), buffer (125 mM Hepes, pH 7.5, 750 mM NaCl, 10 mM MgCl2; 5 μL), purification water (35 μL). After incubation at room temperature for 5 minutes and 30 minutes, 5uL of 5% formic acid was added to stop the reaction. After centrifugation at 15,000 rpm for 10 minutes, the mixture was transferred to LC-MS for detection and data analysis. The parameters of LC and MS are shown in Table 2 and Table 3, respectively. shown.

Table 1 UPLC conditions

Table 2 LC time gradient settings

time (min) A(%) B(%) 0 95 5 0.75 95 5 1 75 25 6 50 50 6.25 0 100 7.5 0 100 7.75 95 5 9 95 5

Table 3 TOFMS parameters are as follows:

Calculate the percent binding of the test compound to the KRASG12C protein:

KRAS G12C binding percentage (%) = peak height of the conjugate of test compound and KRAS G12C protein/[peak height of the conjugate of test compound and KRAS G12C protein + peak height of free KRAS G12C protein] X100. The specific biological analysis data are shown in Table 4.

In-cell Western Blot detection of ERK phosphorylation in H358 cells

H358 cells were recovered and pre-cultured for 3 days until the cells were in good condition (RPMI1640+10%FBS+1%P/S). The cells were seeded into 384-well plates, and the test compound, positive control compound (AMG510 and its isomers) and negative control were added, the compound concentration was 10000nM to 0.051nM, 3-fold dilution, 37°C, 5% _CO2 and mixed well Incubate; cells were washed with PBS and suspended in methanol, washed once with PBS, added with blocking solution, blocked at room temperature for 1 hour, added with primary antibody mixture (rabbit antipERK, mouse anti GAPDH), incubated at 4°C overnight; washed with PBST 3 times, added with two Anti-mixtures (goat antirabbit 800CW and goat anti mouse 680RD) were incubated at room temperature in the dark; the 384-well plate was inverted and centrifuged at 1000 rpm for 1 minute, and the plate was read by Odyssey CLx fluorescence imaging system to obtain fluorescence values; DMSO and ARS1620 were used to correct the reaction values , the specific calculation method is as follows:

Relative Signal=Signal Value(total channel 800)/Signal Value(total channel 700)

H=Ave(DMSO)

L=Ave (ARS1620)

SD(H)=STDEV(DMSO)

SD(L)=STDEV(ARS1620)

CV%(DMSO)=100*(SD_H/Ave_H)

CV%(ARS1620)=100*SD_L/Ave_L

Z'=1-3*(SD_H+SD_L)/(Ave_H-Ave_L)

Relative pERK=(Sample-Ave_L)/(Ave_H-Ave_L).

Four-parameter fitting algorithm analyzes compound IC ₅₀ , and the specific calculation formula is as follows:

Y=Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope))

X: Log of cpd concentration

Y:Ave(relative pERK)

Top and Bottom: Plateaus in same units as Y

logIC50: same log units as X

HillSlope: Slope factor or Hill slope.

The specific biological analysis data are shown in Table 4.

Table 4 Bioanalytical data

Embodiment 41 of WO2018217651A1 discloses the structure of AMG510, and its structure is as follows:

Claims (4)

1. A compound or a pharmaceutically acceptable salt, tautomer or isotopic label thereof, the compound being any of the following:

2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt, tautomer or isotopic label thereof. 3. the compound described in claim 1 or its pharmaceutically acceptable salt, tautomer or isotopic label, or the pharmaceutical composition described in claim 2, in the preparation of prevention and/or treatment KRAS G12C mediated the application of medicines for diseases. 4. The use according to claim 3, wherein the diseases include lung cancer, pancreatic cancer, pancreatic ductal cancer, colon cancer, rectal cancer, appendix cancer, esophageal squamous cell carcinoma, head and neck squamous cell carcinoma and breast cancer.