CN111377918B

CN111377918B - KRAS inhibitor compound

Info

Publication number: CN111377918B
Application number: CN202010222898.2A
Authority: CN
Inventors: 胡永韩; 李昕; 赵金凤; 吴予川; 刘霄; 陈曦
Original assignee: Suzhou Sinovent Pharmaceuticals Co Ltd
Current assignee: Suzhou Sinoway Pharmaceutical Technology Co ltd
Priority date: 2019-11-29
Filing date: 2020-03-26
Publication date: 2021-03-02
Anticipated expiration: 2040-03-26
Also published as: CN111377918A

Abstract

The invention providesA compound having the structure of formula ii or a pharmaceutically acceptable salt, ester, isomer, solvate, prodrug or isotopic label thereof:

Description

KRAS inhibitor compound

Technical Field

The present invention relates to novel KRAS G12C inhibitor compounds and the use of the inhibitor compounds for preventing or treating KRAS G12C mediated diseases.

Background

The Kirsten rat sarcoma virus gene homolog (KRAS) mutation was first described in the NSCLC gene in 1984 as a membrane-bound protein, localized to the inside of the cell membrane; and the KRAS protein and GDP are combined to have no activity under normal conditions, and when the extracellular growth differentiation factor transmits signals to the KRAS protein, the activity of combining the KRAS protein and GTP is enhanced, so that the protein and GTP are combined to be in an activated state, and a signal system is opened. The growth, proliferation, angiogenesis and other processes of tumor cells require intracellular proteins for signal conduction, KRAS gene is a determinant factor of conductive protein, and KRAS mutant type codes abnormal protein to stimulate and promote the growth and diffusion of malignant tumor cells; and is not affected by the signal of upstream EGFR. KRAS mutation promotes cell proliferation, transformation and anti-apoptosis by activating downstream RAS-RAF-MEK-MAPK, P13K-AKT-mTOR and other cell signal transduction pathways, thereby causing tumorigenesis and tumor development.

As shown by COSMIC statistics, the KRAS gene point mutation rate accounts for about 30% of all human tumors, wherein pancreatic cancer accounts for 90%, colon cancer accounts for 45%, and non-small cell lung cancer accounts for 35%. 80% of KRAS mutations occur at codon 12, resulting in a single amino acid substitution, i.e., glycine (G) to alanine (A), cysteine (C), aspartic acid (D), serine (S), arginine (R) and valine (V), with glycine (G) to cysteine (C) being the most common. KRAS G12C mutant, a major proportion (14%) in lung cancer, especially non-small cell lung cancer; it is also expressed in some patients with colorectal (4%), pancreatic (2%) cancer.

Due to the higher expression of KRAS G12C mutation in tumor patients, patients also develop resistance to other targeted drugs, which is attracting increasing attention of more and more experts and scholars. However, drug development directed at KRAS G12C target inhibitors has been challenged by biochemical complexities, which can be compared to the pronoun of oncology "non-druggable" targets, the pharmaceutical world "peaker", which has not been overcome for thirty years.

The development of new drugs is a rapidly developing field, and the discovery of candidate drugs is accelerated by the technical progress. Among these candidate drugs, not only the pharmacodynamics needs to be evaluated, but also the drug metabolism and kinetic properties are very important new drug screening indicators. The ideal drug needs to have a long duration of drug action and good bioavailability. Every year, a large number of drug candidates are eliminated because of their poor pharmacokinetic parameters and metabolic characteristics. Therefore, the metabolic characteristics and the pharmacokinetic parameters of the candidate drug are important evaluation indexes for determining whether the candidate drug can be used as a drug, and good pharmacokinetic parameters and metabolic characteristics are necessary for lead compounds with development prospects. Therefore, providing a KRAS G12C inhibitor with good pharmacokinetic profile would likely be more effective in vivo for pharmacodynamic effects.

Disclosure of Invention

Problems to be solved by the invention

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

Means for solving the problems

In order to solve the technical problems, the invention provides the following technical scheme:

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:

wherein the content of the first and second substances,

R₁' is selected from unsubstituted or substituted by R₇' substituted C_6-10Aryl and 5-10 membered heteroaryl;

R₂' is selected from unsubstituted or substituted by R₈' substituted C_6-10Aryl and 5-10 membered heteroaryl;

R₃' and R₄' each is independently selected from hydrogen, deuterium, C_1-6Alkyl, or R₃And R₄Are linked to form an unsubstituted or optionally substituted 1-3 substituents selected from deuterium, halogen, hydroxy, C_1-63-7 membered cycloalkyl or 3-7 membered heterocycloalkyl substituted with a substituent of alkyl, or R₃And R₄Form ═ O, ═ S, ═ N-CN or ═ CH₂；

R₅' and R₆' are each independently selected from hydrogen, deuterium, and halogen;

each R₇' and R₈' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C_1-6Alkyl, -NHC_1-6Alkyl, -N (C)_1-6Alkyl radical)₂、C_3-6Cycloalkyl radical, C_1-6Alkoxy radical, C_2-6Alkenyl radical, C_2-6Alkynyl, -COOC_1-6Alkyl, said amino, alkyl, cycloalkyl, alkenyl and alkynyl groups being unsubstituted or substituted with 1 to 3 substituents selected from halogen, hydroxy, amino, acetyl or deuterium atoms;

a is unsubstituted or substituted by R₉' substituted 4-9 membered heterocyclic group, each R₉' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkoxy, said amino, alkyl being unsubstituted or substituted by substituents selected from 1 to 3 halogen, cyano, hydroxy, amino or deuterium atoms.

In a preferred embodiment, R is as defined above₁' is selected from unsubstituted or substituted by R₇' substituted C_6-10Aryl and 5-10 membered heteroaryl, said R₇' at C_6-10Ortho-substitution of the atoms bound to the N atom in aryl and 5-to 10-membered heteroaryl groups, the N atom being

The N atom at position 4 in the ring.

In a preferred embodiment, R₁' is selected from unsubstituted or substituted by R₇' substituted C_6-10Aryl and 5-6 membered heteroaryl, said 5-6 membered heteroaryl comprising 1-3 heteroatoms or heteroatom groups selected from N, O, S (O)_mWherein m is an integer of 0 to 2.

In a preferred embodiment, said C_6-10Aryl is selected from phenyl, naphthyl, tetrahydronaphthyl and 2, 3-indanyl, preferably, C_6-10Aryl is phenyl; the 5-to 10-membered heteroaryl group is selected from the group consisting of thienyl, pyridyl, pyridinyloxy, pyrimidinyl, pyrazinyl, pyridazinyl, pyridonyl, pyrazinonyl, pyrimidonyl, pyridazinonyl, pyrrolyl, pyrazolyl, thiazolyl, 1,2, 3-triazolyl, 1,2, 4-triazolylA group selected from the group consisting of imidazolyl, tetrazolyl, isothiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, naphthyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, quinolinyl, isoquinolinyl, and quinazolinyl, preferably, the 5-to 10-membered heteroaryl group is selected from the group consisting of pyridyl or pyrimidinyl.

In a preferred embodiment, each R is₇' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C_1-6Alkyl radical, C_3-6Cycloalkyl and C_1-6Haloalkyl, preferably, each R₇' is independently selected from hydrogen, deuterium, methyl, CH₂F、CHF₂、CF₃Ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, cyclopropane, cyclobutane, cyclopentane and cyclohexane.

In a preferred embodiment, R₂' is selected from unsubstituted or substituted by R₈' substituted C_6-10Aryl and 5-6 membered heteroaryl, said 5-6 membered heteroaryl comprising 1-3 heteroatoms or heteroatom groups selected from N, O, S (O)_rWherein r is an integer of 0 to 2.

In a preferred embodiment, R₂' is selected from unsubstituted or substituted by R₈' substituted C_6-10Aryl and 5-10 membered heteroaryl; said C is_6-10Aryl is selected from phenyl, naphthyl, tetrahydronaphthyl and 2, 3-indanyl; the 5-to 10-membered heteroaryl group is selected from the group consisting of thienyl, pyridyl, pyridinyloxy, pyrimidinyl, pyrazinyl, pyridazinyl, pyridonyl, pyrazinonyl, pyrimidonyl, pyridazinonyl, 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 is R₂' is selected from unsubstituted or substituted by R₈' substituted phenyl, imidazolyl, pyrrolyl, pyridinyloxy, pyridyl, pyridonyl, naphthyl, quinolinyl, isoquinolinylAnd quinazolinyl.

In a preferred embodiment, each R is₈' are each independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C_1-6Alkyl, -NHC_1-6Alkyl, -N (C)_1-6Alkyl radical)₂、C_1-6Alkoxy, said amino, alkyl, unsubstituted or substituted by 1 to 3 substituents selected from halogen, hydroxy, amino, acetyl or deuterium atoms, preferably, each R₈' is independently selected from hydrogen, deuterium, fluorine, chlorine, hydroxyl and amino.

In a preferred embodiment, R₃' and R₄' independently of each other are selected from hydrogen, deuterium, C_1-6Alkyl, or R₃And R₄Are linked to form a cyclopropyl group, or R₃' and R₄' formation of ═ O, ═ S, or ═ N — CN, preferably, R₃' and R₄Form ═ O.

In a preferred embodiment, R₅' and R₆' are each independently selected from hydrogen, deuterium, fluorine and chlorine.

In a preferred embodiment, A is unsubstituted or is each independently substituted by R₉' A substituted 4-9 membered heterocyclic group, and the atom to which the 4-9 membered heterocyclic group is attached to the carbonyl group is N.

In a preferred embodiment, A is unsubstituted or substituted with R₉' substituted 4-9 membered heterocyclic group, said 4-9 membered heterocyclic group includes monocyclic, fused ring, bridged ring, spiro ring.

In a preferred embodiment, A is unsubstituted or substituted by 1 to 3R₉' substituted 6-7 membered heterocyclic group, said 6-7 membered heterocyclic group containing no double bond or 1 or 2 double bonds, preferably, A is unsubstituted or substituted with 1-2R₉' substituted 6-7 membered heterocycle, said 6-7 membered heterocycle selected from

In a preferred embodiment, each R is₉' 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, R₁' is selected from unsubstituted or substituted by R₇' substituted C_6-10Aryl and 5-10 membered heteroaryl;

R₃' and R₄' each is independently selected from hydrogen, deuterium, C_1-6Alkyl, or R₃And R₄Are linked to form a cyclopropyl group, or R₃And R₄To form ═ O;

each R₇' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C_1-6Alkyl radical, C_3-6A cycloalkyl group;

each R₈' is independently selected from hydrogen, deuterium, cyano, halogen, hydroxy, amino, C_1-6Alkyl, -NHC_1-6Alkyl, -N (C)_1-6Alkyl radical)₂、C_1-6Alkoxy, said amino, alkyl, unsubstituted or substituted with 1 to 3 halogen, hydroxyl, amino, acetyl or deuterium atoms;

a is unsubstituted or substituted by 1 to 3R₉' A substituted 6-7 membered heterocyclic ring, and the atom to which the 6-7 membered heterocyclic ring is attached to the carbonyl group is N, each R₉' is independently selected from hydrogen, deuterium, methyl, ethyl, -CH₂OH and-CH₂F。

In a more preferred embodiment, R₁' is selected from unsubstituted or substituted by R₇' substituted C_6-10Aryl and 5-10 membered heteroaryl; said C is_6-10Aryl is selected from phenyl, naphthyl, tetrahydronaphthyl and 2, 3-indanyl; said 5-to 10-membered heteroaryl group is selected fromThienyl, pyridyl, pyridinyloxy, pyrimidinyl, pyrazinyl, pyridazinyl, pyridonyl, pyrazinyl, pyrimidinonyl, pyridazinyl, 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;

R₂' is selected from unsubstituted or substituted by R₈' substituted C_6-10Aryl and 5-10 membered heteroaryl; said C is_6-10Aryl is selected from phenyl, naphthyl, tetrahydronaphthyl and 2, 3-indanyl; the 5-to 10-membered heteroaryl group is selected from the group consisting of thienyl, pyridyl, pyridinyloxy, pyrimidinyl, pyrazinyl, pyridazinyl, pyridonyl, pyrazinonyl, pyrimidonyl, pyridazinonyl, 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;

R₃' and R₄' form ═ O;

R₅' and R₆' each is 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 and cyclohexane;

each R₈' is independently selected from hydrogen, deuterium, fluorine, chlorine, hydroxyl and amino;

a is unsubstituted or substituted by 1 to 2R₉' substituted 6-7 membered heterocyclic ring, said 6-7 membered heterocyclic group being 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, which is any one of:

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

In addition, the invention also provides application of the compound or the pharmaceutically acceptable salt, ester, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotopic marker or prodrug thereof, or the pharmaceutical composition in preparing a medicament for preventing and/or treating KRAS G12C-mediated diseases, preferably lung cancer, pancreatic duct cancer, colon cancer, rectal cancer, appendiceal cancer, esophageal squamous carcinoma, head and neck squamous carcinoma, breast cancer and other solid tumors.

In order to more clearly describe the context of the present invention, the terms referred to will now be defined as follows:

in the present invention, the term "C_1-6Alkyl "represents, alone or in combination, a saturated, linear or branched alkyl group containing from 1 to 6, in particular from 1 to 4, carbon atoms, including methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-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-pentyl, 2, 3-dimethyl-2-butyl, 3, -dimethyl-2-butyl, and the like. Preference is given toGround,' C_1-6The alkyl group "is any of methyl, ethyl, n-propyl, isopropyl, and tert-butyl. Similarly, the term "C_1-3Alkyl "alone or in combination means a saturated straight or branched chain alkyl group containing 1 to 3 carbon atoms, including methyl, ethyl, propyl, isopropyl, and the like.

The term "3-7 membered cycloalkyl" alone or in combination means cycloalkyl having 3 to 7, especially 3-6 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. In particular "C_3-7Cycloalkyl "is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term "amino" denotes, alone or in combination, a primary amino group (-NH)₂) Secondary amino (-NH-) or tertiary amino

The term "C_1-6Alkoxy "alone or in combination denotes the radical C_1-6alkyl-O-in which "C_1-6Alkyl represents as defined above, including but not limited to methoxy (-OCH)₃) Ethoxy (-OCH)₂CH₃) N-propoxy group (-OCH)₂CH₂CH₃) I-propoxy (-OCH (CH)₃)₂) N-butoxy (-OCH)₂CH₂CH₂CH₃) Sec-butoxy (-OCH (CH)₃)CH₂CH₃) Isobutoxy (-OCH)₂CH(CH₃)₂) T-butoxy (-OC (CH))₃)₃) N-pentyloxy (-OCH)₂CH₂CH₂CH₂CH₃) Neopentyloxy (-OCH)₂C(CH₃)₃) And the like.

The term "halogen" alone or in combination means fluorine, chlorine, bromine or iodine. In particular fluorine, chlorine or bromine.

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

The term "4-9 membered heterocyclyl" means a monocyclic, fused, bridged, spiro ring containing 4 to 9, especially 6 to 7 carbon atoms and a heteroatom or heteroatom group selected from N, O, S (O) which is free of double bonds or contains 1 or 2 double bonds_m(wherein m is an integer of 0 to 2); for example azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuryl, tetrahydrothienyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, tetrahydropyranyl, 1-dioxothiomorpholinyl, bicyclo [4.1.0 ] and containing no or 1 or 2 double bonds]Heptyl, and the like.

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

The term "heteroaryl" denotes an aromatic ring group formed by replacement of a carbon atom on the ring by at least one heteroatom or heteroatom group selected from N, O, S (O)_m(wherein m is an integer of 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, pyridinyloxy (i.e. pyridinyloxy)

) Pyridonyl, pyrazinonyl, pyrimidinonyl, pyridazinonyl, pyrrolyl, pyrazolyl, thiazolyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, imidazolyl, tetrazolyl, isothiazolyl, oxazolylOxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, quinolinyl, isoquinolinyl, quinazolinyl, and the like. The hydrogen atoms on the "heteroaryl" groups are independently optionally substituted with one or more substituents as described herein.

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

The term "C_6-10Aryl "denotes an aryl group having 6 to 10 carbon atoms, wherein aryl denotes 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" encompasses all isomeric forms including enantiomers, diastereomers, tautomers and geometric isomers (including cis-trans isomers). Thus, individual stereochemical isomers of the contemplated compounds of the present invention or mixtures of enantiomers, diastereomers, tautomers or geometric isomers (or cis-trans isomers) thereof are intended to be 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 pharmaceutical salts described in J.pharmaceutical Sciences (Vol.66: pp.1-19, 1977) by S.M.Berge. In the present invention, pharmaceutically acceptable non-toxic acid addition salts mean salts of the compounds of the present invention with organic or inorganic acids including, but not limited to, hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic 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 acidToluene sulfonic acid, malic acid, and the like. Pharmaceutically acceptable non-toxic base addition salts mean salts of the compounds of the 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; salts of organic bases, e.g. ammonium salts formed by reaction with organic bases containing N groups or N⁺(C_1-6Alkyl radical)₄The salt is preferably lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogencarbonate, potassium carbonate, potassium hydrogencarbonate, magnesium carbonate, calcium carbonate, ammonia water, triethylamine, tetrabutylammonium hydroxide, or the like.

The term "solvate" refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, N-dimethylformamide, dimethylsulfoxide, and the like. The "pharmaceutically acceptable salt" can be synthesized by a general chemical method.

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

The term "hydrate" refers to an association of water with a compound of the present invention.

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

The term "isotopic derivative" means an isotopic derivative wherein the hydrogen atom in formula (I) is replaced by 1 to 6 deuterium atoms (D), the carbon atom in formula (I) is replaced by 1 to 3 carbon 14 atoms (I)¹⁴C) The resulting isotopic derivative by substitution.

The terms related to the present invention are defined above, and those skilled in the art can also understand the above terms in combination with the prior art, and further description is made below based on the contents of the present invention and the definitions of the terms.

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

Detailed Description

Example 1SZ-014096

The first step is as follows: synthesis of compound 014004A2

Compound 2, 6-dichloro-5-fluoronicotinic acid 014004a1(16.0g, 76.4mmol) was dissolved in dichloromethane (200mL), oxalyl chloride (12.1g, 95.2mmol) was added dropwise in an ice-water bath, a catalytic amount of DMF (0.2mL) was then added dropwise, the reaction was allowed to react overnight at room temperature (20 ℃), the resulting residue was concentrated under reduced pressure, the resulting residue was dissolved in 200mL1, 4-dioxane and cooled to zero, an aqueous ammonia solution (28.0-30%, 14.4mL, 114.2mmol) was added dropwise slowly to the reaction phase, the reaction was stirred at zero temperature for 30 minutes, the solvent was removed by concentration under reduced pressure, and the resulting crude white solid was purified by silica gel column chromatography (dichloromethane/methanol 100: 1) to give compound 014004a2 as a white solid (9.0g, yield 56.2%). LCMS (M + H)⁺m/z calculated 209.0, found 209.0.

The second step is that: synthesis of compound 014004A4

To a suspension (60mL) of the compound 2-bromo-4-methyl-3-aminopyridine 014004A3(5.4g, 28.9mmol) in tetrahydrofuran was added Pd (dppf) Cl₂(1.02g, 1.4mmol), isopropyl magnesium chloride (2M in tetrahydrofuran, 22mL, 43.4mmol) was added to the reaction mixture at zero degrees under nitrogen, and after addition, the reaction was stirred at 60 degrees overnight. The reaction was stopped and cooled to room temperature, the reaction was quenched with a mixture of 100mL water and 200mL 1N sodium hydroxide solution, extracted with ethyl acetate (200mL × 2), the combined organic phases were concentrated under reduced pressure, and the concentrated residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 1: 1) to give 014004a4 as a yellow colloidal compound (3.22 g, yield 73.1%). LCMS (M + H)⁺m/z calculated 151.1, found 151.1.¹H NMR(DMSO-d₆，400MHz)：δ8.24(d，J＝8.0Hz，1H)，8.11(br s，1H)，7.95(br s，1H)，2.17(s，3H)。

The third step: synthesis of compound 014004A5

Compound 014004A2(5.0g, 23.9mmol) was dissolved in tetrahydrofuran (20mL) and oxalyl chloride (3.66g, 28.8mmol) was added slowly under an ice-water bath. The reaction mixture was stirred at 75 ℃ for 1 hour. The heating was stopped and the temperature was reduced to zero, compound 014004A4(3.6g, 23.9mmol) in tetrahydrofuran (10mL) was slowly added dropwise, and the reaction was stirred with zero for an additional 1 hour, followed by 1: 1, extracted with ethyl acetate (200mL x 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give residue 014004a5(9.1g, crude) which was used in the next step without purification. LCMS (M + H)⁺m/z calculated 385.1, found 385.1.

The fourth step: synthesis of compound 014004A6

Crude compound 014004a5(9.1g, 23.8mmol) was dissolved in tetrahydrofuran (40mL), KHMDS (1M in THF, 50.2mL, 50.2mmol) was slowly added dropwise under ice bath, the ice bath was removed, the reaction was stirred at room temperature (20 ℃) for 1 hour, the reaction phase was quenched with saturated ammonium chloride solution, ethyl acetate was extracted (100mL × 3), the organic phase was combined and dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to give a residue which was purified by silica gel column chromatography (petroleum ether/ethyl acetate/dichloromethane/methanol: 100: 10: 10:: 1) to give 014004a6(3.2g, yield 38.2%) as a pale yellow solid. LCMS (M + H)⁺m/z calculated 349.1, found 349.1.¹H NMR(DMSO-d₆，400MHz)：δ12.27(br s，1H)，8.53-8.49(m，2H)，7.29(d，J＝4.8Hz，1H)，2.87(q，J＝6.6Hz，1H)，2.05-1.99(m，3H)，1.09(d，J＝6.6Hz，3H)，1.01(d，J＝6.6Hz，3H)。

The fifth step: synthesis of compound 014004A7

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

And a sixth step: 014004A8 Synthesis

Crude compound 014004A7(2.2g, 6.0mmol) was dissolved in acetonitrile (10mL), N-diisopropylethylamine (2.3g, 18.0mmol) and (S) -4-N-tert-butoxycarbonyl-2-methylpiperazine (1.44g, 7.2mmol) were added under ice-bath, the ice-bath was removed, the reaction was stirred at room temperature (20 ℃ C.) for 1 hour, ice saturated sodium bicarbonate solution (100mL) and ethyl acetate (150mL) were then added, stirring was continued for 5 minutes, the phases were separated, the aqueous phase was further extracted with ethyl acetate (100mL × 2), the organic phase was combined and dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to give the residue which was purified by silica gel column chromatography (petroleum ether/ethyl acetate/dichloromethane/methanol 80: 10: 10:: 1) to give compound 014004a8(1.8g, 56.3% yield) as a pale yellow solid. LCMS (M + H)⁺m/z calculated 531.2, found 531.2.¹H NMR(CDCl₃，400MHz)：δ8.56(d，J＝4.8Hz，1H)，7.78(d，J＝7.2Hz，1H)，7.13(d，J＝4.8Hz，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)。

The seventh step: 014004A9 Synthesis

Compound 014004a8(1.7g, 3.21mmol), 2-fluoro-6-hydroxyphenylboronic acid (605.6mg, 3.91mmol), potassium acetate (1.62g, 16.5mmol) and pd (dppf) Cl2(119mg, 0.17mmol) were dissolved in 1, 4-dioxane (25mL), after several nitrogen replacements, the reaction was stirred at 90 ℃ for 1 minute, followed by 2 drops of water, stirring at 90 ℃ for 1 hour, cooling to room temperature, water (25mL) and brine (25mL) were added to the reaction phase, extracted with ethyl acetate (200mL × 2), the organic phase was combined and dried over anhydrous sodium sulfate, and the filtrate was concentrated to give the residue which was purified by silica gel column chromatography (petroleum ether/ethyl acetate/dichloromethane/methanol 60: 10: 10:1) to give 014004a9(1.6g, 82.5% yield) as a yellow solid.

Followed by chiral preparation (CO) by supercritical chromatography₂EtOH 60/40) gave the yellow solid compound 014004A9A (elution isomer 1) (700mg) and014004A9B (eluted isomer 2) (650 mg). LCMS (M + H)⁺m/z calculated 607.3, found 607.3.

Chiral preparation conditions are as follows: column: IC (integrated circuit)

*250mmL*5um；Method：CO₂:EtOH＝60/40

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

Eighth step: 014004A10A and 014004A10B Synthesis

Compound 014004A9A (700mg, 1.15mmol) was dissolved in dichloromethane (6mL), trifluoroacetic acid (2.8mL, 35mmol) was added under ice bath and the ice bath was removed, the reaction was stirred at room temperature (20 ℃) for 1 hour, the reaction was dried by spinning, to the resulting brown oily residue was added ice saturated sodium bicarbonate solution (10mL), extracted with dichloromethane (100mL × 2), and washed with water (50mL × 2). The organic phase was dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to give residue 014004A10A (550mg, crude, 94.2% yield) which was used in the next step without purification. LCMS (M + H)⁺m/z calculated 507.2, found 507.2.

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

The ninth step: 014096A1 Synthesis

2-Methylfuran (2.0g, 24.4mmol) was dissolved in tert-butanol-water (5:1, v/v,120mL), sodium dihydrogen phosphate (5.7g, 36.6mmol) sodium chlorite (6.6g, 73.2mmol) was added with stirring, and the reaction phase was stirred at room temperature (25 ℃ C.) for two hours or until the yellow color disappeared. The reaction solution was removed by rotary evaporation, and the resulting residue was extracted with chloroform (300ml) and washed with brine (2)200ml), the organic phase was dried over magnesium sulfate, filtered and the solvent was spin-dried, and the resulting yellow oily residue 014096a1 was used in the next reaction without purification. (300mg, crude yield 10.8%). LCMS (M + H)⁺m/z calculated 115.0, found 115.1.

The tenth step: 014096A2 Synthesis

Compound 014096a1(300mg, 2.63mmol) was dissolved in tetrahydrofuran-acetone-water (5: 4: 1, 40ml), freshly distilled pyridine (1 mol%, 200uL) was added, the reaction phase was stirred at room temperature (25 ℃) for two hours, and the crude oil obtained by concentrating the reaction solution was purified by silica gel column chromatography (ethyl acetate/petroleum ether ═ 1: 2) to give crude compound 014096a2(80mg, yield 26.7%) as a yellow oil. LCMS (M + H)⁺m/z calculated 115.0, found 115.1.

The eleventh step: SZ-014096 Synthesis

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

Liquid phase mass spectrometry [ mobile phase: elution was carried out in a gradient from 70% water (containing 0.02% ammonium acetate) and 30% acetonitrile to 50% water (containing 0.02% ammonium acetate) and 50% acetonitrile at a flow rate of 1.5 ml/min for 6.5 minutes at a column temperature of 40 ℃. Column: waters XBridge C183.5um, 50x4.6mm]Purity 96.76%, Rt 2.889 min; LCMS (M + H)⁺m/z calculated 603.3, found 603.3.¹H NMR(DMSO-d₆，400MHz)：δ10.20(br s，1H)，8.39(d，J＝4.8Hz，1H)，8.27(dd，J₁＝21.6Hz，J₂＝9.2Hz，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＝6.4Hz，3H)，0.93(d，d，J＝6.8Hz，3H)。

Example 2SZ-014096B

The first step is as follows: SZ-014096B Synthesis

Compound 014004A10B (100mg, 0.2mmol),014096A2(22mg, 0.2mmol), HATU (112mg,0.296mmol) was dissolved in N, N-dimethylformamide (2.0mL), to which N, N-diisopropylethylamine (51mg, 0.4mmol) was added. The reaction phase was stirred at room temperature (25 ℃ C.) for two hours. Concentration under reduced pressure removed N, N-dimethylformamide, and the concentrated residue was purified by preparative high performance liquid chromatography to give SZ-014096B (23.0mg, yield 19.3%) as a white solid. Liquid phase mass spectrometry [ mobile phase: elution was carried out in a gradient from 80% water (containing 0.02% ammonium acetate) and 20% acetonitrile to 30% water (containing 0.02% ammonium acetate) and 70% acetonitrile at a flow rate of 1.5 ml/min for 6.5 minutes at 40 degrees celsius column temperature. Column: waters XBridge C183.5um, 50x4.6mm]Purity 99.14%, Rt 3.361 min; LCMS (M + H)⁺m/z calculated 603.3, found 603.2.¹H NMR(DMSO-d₆，400MHz)：δ10.18(br s，1H)，8.39(d，J＝4.8Hz，1H)，8.30(dd，J₁＝19.6Hz，J₂＝9.2Hz，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＝6.0Hz，3H)。

Example 3SZ-014017A/B

The first step is as follows: 014086A1 Synthesis

The compound 4, 6-dichloro-5-aminopyrimidine (10.0g, 60.98mmol) was dissolved in 200ml of tetrahydrofuran, and [1,1' -bis (diphenylphosphino) ferrocene was added]Palladium dichloride (8.9g, 12.2mmol), isopropyl magnesium chloride (1N, 183mL, 366mmol) was added dropwise at zero degrees and heated to 70 degrees overnight under nitrogen. The reaction solution was cooled to room temperature, quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and 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 014086a1(3.0g, yield 27%) as a yellow solid. LCMS (M + H)⁺m/z calculated 180.1, found 180.1.¹H NMR(DMSO-d₆，400MHz)：δ8.34(s，1H)，5.06(s，2H)，3.29-3.20(m，2H)，1.17(d,J＝8.8Hz，12H)。

The second step is that: 014086A2 Synthesis

The compound 2, 6-dichloro-3-carboxamide-5-fluoropyridine (3.0g, 14.4mmol) was dissolved in 50ml of tetrahydrofuran, oxalyl chloride (2.2g, 17.28mmol) was added thereto at room temperature, and the reaction was refluxed for 1 hour. The reaction was cooled to zero, and compound 014086A1(2.6g, 14.4mmol) was added and 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 014086a2(5.6g, yield 90%) as a yellow solid. LCMS (M + H)⁺m/z calculated 414.2, found 414.2.

The third step: 014086A3 Synthesis

Compound 014086A2(5.2g, 12.56mmol) was dissolved in 50mL of anhydrous tetrahydrofuran, KHMDS (1N, 27.63mL) was added dropwise over an ice bath, and the reaction was stirred at room temperature for 1 hour. The reaction solution was poured into water, neutralized 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) to obtain 014086A3(4.3g, yield 89%) as a pale yellow solid. LCMS (M + H)⁺m/z calculated 378.2, found 378.2.¹H NMR(DMSO-d₆，400MHz)：δ12.36(s，1H)，9.22(s，1H)，8.55(d，J＝10.0Hz，1H)，3.00-2.96(m，2H)，1.12(d，J＝8.8Hz，6H)，1.02(d，J＝8.8Hz，6H)。

The fourth step is 014086A4 synthesis

Compound 014086A3(800mg, 2.02mmol) was dissolved in 5mL of anhydrous acetonitrile and POCl was added under ice-bath₃(487mg, 3.18mmol) and DIPEA (546mg, 4.24mmol), and the reaction was heated to 80 ℃ for reaction overnight. The reaction was spun dry to give 014086A4 as a brown oil (800mg, 100% yield) which was used directly in the next step.

The fifth step: 014086A5 Synthesis

Crude compound 014086A4(800mg, 2.02mmol) was dissolved in 10ml of acetonitrile, and (S) -4-N-tert-butoxycarbonyl-2-methylpiperazine (404mg, 2.02mmol) and DIPEA (781.74mg, 6.06mmol) were added and stirred at room temperature for 1 hour. The reaction was spun dry and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 3:1) to give 014086a5(600mg, yield 54.5%) as a pale yellow solid. LCMS (M + H)⁺m/z 560.2, found 560.2.¹H NMR(DMSO-d₆，400MHz)：δ9.15(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)。

And a sixth step: 014086A6 Synthesis

Compound 014086A5(560mg, 1mmol), 2-fluoro-6-hydroxyphenylboronic acid (1.24g, 8mmol) were dissolved in 10ml of 1, 4-dioxane, and potassium acetate (490mg, 5mmol) and [1,1' -bis (diphenylphosphino) ferrocene were added]Palladium dichloride (146mg, 0.2mmol) was reacted for 90 minutes at 120 ℃ with a microwave. The reaction was cooled, filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 3:1) to give 014086a6(300mg, yield 47%) as a pale yellow solid. LCMS (M + H)⁺m/z calculated 636.3, found 636.3.

The seventh step: 014086A7 Synthesis

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

Eighth step: SZ-014017A/B Synthesis

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

Liquid phase mass spectrometry [ mobile phase: elution was carried out in a gradient from 70% water (containing 0.02% ammonium acetate) and 30% acetonitrile to 40% water (containing 0.02% ammonium acetate) and 60% acetonitrile at a flow rate of 1.5 ml/min for 6.5 minutes at 40 degrees celsius column temperature. Column: waters XBridge C183.5um, 50x4.6mm]Purity 99.14%, Rt 3.129 min; LCMS (M + H)⁺m/z calculated 632.2, found 632.2.¹H NMR(DMSO-d₆，400MHz)：δ10.20(br s，1H)，9.05(s，1H),8.32(dd，J₁＝22.0Hz，J₂＝9.2Hz，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＝6.8Hz，3H)，1.08(d，J＝6.8Hz，6H)，0.932(d，J＝6.4Hz，6H)。

Biological activity assay

LC-MS method for detecting KRAS G12C protein binding rate

Test compounds were prepared as 10mM stock solutions in DMSO. KRAS G12C protein was dissolved in buffer (20mM Hepes, pH7.5, 50mM NaCl, 0.5mM MgCl)₂) Diluted to 103uM, GDP buffer (20mM Hepes, pH7.5, 50mM NaCl, 0.5mM MgCl) was added in equal volume₂10mM EDTA, 2mM DTT, GDP) was prepared as KRASG12C protein loaded with GDP.

The KRASG12C protein loaded with GDP was added to a dilution solution (12.5mM Hepes, ph7.5, 75mM NaCl,10mM MgCl₂) Dilute to 20 uM. The reaction system is prepared from the following components: GDP-KRAS-4B-G12C (20uM, 5. mu.L), test compounds (10% DMSO, 5. mu.L), buffer (125mM Hepes, pH7.5, 750mM NaCl, 10mM MgCl 2; 5. mu.L), purified water (35. mu.L). After incubation at room temperature for 5 min and 30 min, 5uL of 5% formic acid was added to stop the reaction, after centrifugation at 15000rpm for 10 min, the mixture was transferred to LC-MS for detection and data analysis, the parameters of LC and MS are shown in tables 2 and 3, respectively.

TABLE 1 UPLC conditions

TABLE 2 LC time gradient setup

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:

the percent binding of the test compound to KRASG12C protein was calculated:

KRAS G12C binding percentage (%): test compound peak height with KRAS G12C protein conjugate/[ test compound peak height with KRAS G12C protein conjugate + free KRAS G12C protein peak height ] X100. Specific bioanalytical data are shown in table 4.

H358 cell In-cell WesternBlot detection of ERK phosphorylation

H358 cells were revived and pre-cultured for 3 days to a good cell status (RPMI1640+ 10% FBS + 1% P/S). Cells were seeded into 384-well plates and test compounds, positive control compounds (AMG510 and isomers thereof) and negative controls were addedThe compound concentration is 10000nM to 0.051nM, 3 times dilution, 37 deg.C, 5% CO₂Mixing evenly and incubating; washing cells with PBS and suspending with methanol, washing with PBS again, adding blocking solution, blocking at room temperature for 1 hr, adding primary antibody mixture (rabbitanti pERK, mouse anti GAPDH), and incubating at 4 deg.C overnight; PBST was washed 3 times, and a secondary antibody mixture (coat anti rabbit800 CW and coat anti mouse 680RD) was added and incubated at room temperature in the dark; the 384-well plate is reversely centrifuged at 1000rpm for 1 minute, and the Odyssey CLx fluorescence imaging system reads the plate to obtain the fluorescence value; the reaction values were corrected using DMSO and ARS1620, calculated 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 for analysis of compound IC₅₀The 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。

specific bioanalytical data are shown in table 4.

TABLE 4 biological assay data

Example 41 of WO2018217651a1 discloses the structure of AMG510 as follows:

Claims

1. a compound, or a pharmaceutically acceptable salt, tautomer, or isotopic label thereof, which compound is any one of:

2. a pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or isotopic label thereof.

3. Use of a compound of claim 1 or a pharmaceutically acceptable salt, tautomer or isotopic label thereof, or a pharmaceutical composition of claim 2, in the manufacture of a medicament for the prevention and/or treatment of a KRAS G12C-mediated disease.

4. The use of claim 3, wherein the disease is selected from the group consisting of lung cancer, pancreatic ductal cancer, colon cancer, rectal cancer, appendiceal cancer, esophageal squamous cancer, head and neck squamous cancer, and breast cancer.