Kras g12c mutant protein inhibitors

Abstract

The application discloses a compound for irreversibly inhibiting KRAS G12C mutation, a pharmaceutically acceptable salt of the compound and a pharmaceutical composition containing the compound or the salt thereof, and further discloses application of the compound or the salt thereof and the pharmaceutical composition in treating proliferative diseases such as KRAS G12C mutant tumors.

Description

TECHNICAL FIELD

[0001] The present application relates to a compound irreversibly inhibiting KRAS G12C mutation, a pharmaceutically acceptable salt of the compound and a pharmaceutical composition containing the compound or the salt thereof, and also relates to the use of the compound or the salt thereof and the pharmaceutical composition in treating a proliferative disease such as tumor caused by KRAS G12C mutation. BACKGROUND

[0002] RAS proteins can be divided into KRAS, HRAS and NRAS according to their amino acid sequences. RAS proteins enter "activated" or "inactivated" states by binding to guanine triphosphate (GTP) or guanine diphosphate (GDP), respectively. RAS proteins bind to GDP in resting cells, making RAS in an inactivated state; when cells are activated, RAS proteins bind to GTP to form GTP-RAS, and at the same time activate RAS and its downstream signals (Nature Review Cancer 3:11-22, 2003). When RAS proteins are mutated, due to the increase of activated GTP-RAS, RAS signals are in a persistent activated state, continuously activating downstream signals, stimulating abnormal cell proliferation, and inducing tumor occurrence.

[0003] KRAS, HRAS and NRAS can all be mutated, but K-RAS (Kirsten rat sarcoma onco gene) is the most frequently mutated oncogene in tumors. The most common K-RAS mutations occur at the 12th glycine (G12), the 13th glycine (G13) and the 61st glutamine (Q61) residues; among them, the mutation at the G12 position has the highest incidence (Nat Rev Drug Discov 2014, 13:828-851). K-RAS G12C mutation refers to the mutation of glycine at the 12th position of K-RAS protein to cysteine, which is the most common type of K-RAS mutation. The occurrence frequency of K-RAS G12C mutant tumors is in turn pancreatic cancer (57%), colorectal cancer (35%), biliary tract cancer (28%), small intestine cancer (17%), lung cancer (16%), endometrial cancer (15%) and ovarian cancer (14%) and the like (Seminars in Cancer Biology. 2019 Jun 27. pii: S1044-579X(18)30060-9). Malignant tumors with K-RAS G12C mutation are not sensitive to conventional treatment, so patients have poor prognosis and short survival time.

[0004] The discovery of the RAS oncogene has a history of more than 30 years, and the pharmaceutical industry has high expectations for RAS inhibitors, especially anti-K-RAS G12C mutant RAS inhibitors. K-RAS G12C mutant protein inhibitors are a new type of drug target for anti-RAS targeted therapy discovered in recent years (Nature 503:548-551, 2013).

[0005] K-RAS G12C covalent inhibitors are designed to utilize the nucleophilic reactivity of the 12th cysteine after mutation; by modifying with a disulfide bond, the compound enters the K-RAS G12C allosteric pocket, and by modifying with a disulfide bond, it blocks the activation of K-RAS G12C mutant protein to inhibit tumor growth. However, so far no new K-RAS G12C inhibitor with high activity and high safety has been approved by regulatory agencies (Cell Chem Biol. 2019, 26(10): 1338-1348). Therefore, there is still an urgent need to discover new highly selective K-RAS G12C inhibitors for the molecular targeted therapy of K-RAS G12C mutant tumors (Cancer Treat Rev. 2020, 84:101974). SUMMARY

[0006] To solve the above technical problems, the present application provides new compounds for inhibiting KRAS G12C mutant proteins.

[0007] In one aspect, the present application provides a compound represented by Formula I or Formula II or Formula III, an isomer thereof, a pharmaceutically acceptable salt thereof, or a deuterated product thereof:

[0008]

[0009] wherein:

[0010] W is a 4-12 membered saturated or partially saturated monocyclic, bicyclic, bridged or spiro ring, wherein the saturated or partially saturated monocyclic, bicyclic, bridged or spiro ring is optionally substituted with one or more R2;

[0011] P and K are nitrogen or carbon, preferably P and K can be both nitrogen, or at least one of P and K is nitrogen;

[0012] B is absent or -NH-; when B is absent, P is directly connected to R1;

[0013] R1 is

[0014]

[0015] E is CR7 or NR8;

[0016] G is CR9 or NR 10 ;

[0017] Q is CR 11 NR 12 N or C = O;

[0018] Is it a single bond or a double bond?

[0019] R2 is a C1-C3 alkyl, C2-C4 alkynyl, heteroalkyl, cyano or oxo, wherein the C1-C3 alkyl may be optionally substituted with halogen, cyano or heteroaryl.

[0020] R3 is absent, hydrogen, halogen, CF3, hydroxyalkyl, or C1-C3 alkyl;

[0021] R4 is absent, hydrogen, C1-C3 alkyl, alkylaminoalkyl, heterocyclic, aminoalkyl, halogen, amide, nitrile, hydroxyalkyl, CF3, CF2, methoxy, trifluoromethyl, amino or alkenyl;

[0022] R5 is absent, hydrogen, alkyl, or hydroxyalkyl;

[0023] R6 is absent, hydrogen, or a C1-C3 alkyl group;

[0024] R7 is hydrogen, C1-C4 alkyl, cycloalkyl, amino, cyano, alkynyl, halogen, oxo, aminoalkyl, alkylaminoalkyl, or haloalkyl.

[0025] R8 is hydrogen, C1-C4 alkyl, cycloalkyl, alkynyl, or -CONH2;

[0026] R9 is a cycloalkyl, heterocyclic, aryl, aralkyl, or heteroaryl group; the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups may be optionally combined to form monocyclic, bicyclic, or tricyclic groups; each of the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups may be represented by one or more R9 groups. 13 replace;

[0027] R 10 It is a cycloalkyl, heterocyclic, aryl, aralkyl, or heteroaryl group; the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups can be optionally combined to form monocyclic, bicyclic, or tricyclic groups; each of the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups can be represented by one or more R groups. 15 replace;

[0028] R 11 It is absent, hydrogen, C1-C4 alkyl, cycloalkyl, amino, or halogen;

[0029] R 12 It is hydrogen, C1-C4 alkyl, cycloalkyl, or alkynyl;

[0030] R 13 It is a halogen, hydroxyl, C1-C6 alkyl, cycloalkyl, alkoxy, haloalkyl, amino, cyano, heteroalkyl, hydroxyalkyl, oxoalkyl, or thioalkyl;

[0031] R 14 It is hydrogen, alkyl, aminoalkyl, alkylaminoalkyl, haloalkyl, hydroxyalkyl, dihydroxyalkyl, 4-6 membered heterocyclic, 4-6 membered heterocyclic alkyl, 5-6 membered aryl, 5-6 membered heteroaryl or heteroarylalkyl, wherein the heterocyclic, heterocyclic alkyl, aryl, heteroaryl or heteroarylalkyl may be optionally surrounded by one or more identical or different R 15 replace;

[0032] R 15 It is a halogen, hydroxyl, hydrogen, C1-C6 alkyl, cycloalkyl, alkoxy, haloalkyl, amino, cyano, heteroalkyl, oxoalkyl, aralkyl, or acyl.

[0033] R 17 It is a cycloalkyl, heterocyclic, aryl, aralkyl, or heteroaryl group; each of the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups may optionally be converted by one or more R groups. 16 replace;

[0034] R 16 It is hydrogen, C1-C4 alkyl, amino, cyano, alkynyl, halogen, oxo, aminoalkyl, or haloalkyl;

[0035] R 18 It is hydrogen, C1-C4 alkyl, cycloalkyl, amino, halogen, or trifluoromethyl;

[0036] J is either O or S.

[0037] This invention also includes compounds of formulas I-1, I-2, I-3, I-4, I-5, II-1, II-2, II-3, and III-1, their isomers, pharmaceutically acceptable salts thereof, or their deuterated derivatives:

[0038] When E is NR8, G is CR9, and Q is CR in equation I 11 Then, we get Equation I-1;

[0039] When E is CR7 and G is NR in equation I 10 When Q is C=0, we get Equation I-2;

[0040] When E is NR8, G is CR9, and Q is N in Equation I, we get Equation I-3;

[0041] When E is CR7 and G is NR in equation I 10 Q is CR 11Then, we get Equation I-4;

[0042] When E is CR7, G is CR9, and Q is NR in equation I 12 Then, we obtain Equation I-5;

[0043] When E is NR8, G is CR9, and Q is CR in Equation II 11 Then, we obtain Equation II-1;

[0044] When E is CR7 and G is NR in Equation II 10 Q is CR 11 Then, we obtain Equation II-2;

[0045] When E is CR7, G is CR9, and Q is NR in Equation II 12 Then, we obtain Equation II-3;

[0046] When E is CH and G is NR in Equation III 10 When Q is C=O, we get Equation III-1;

[0047]

[0048]

[0049] Among them, R1, R2, R7, R8, R9, R 10 R 11 R 12 R 14 R 16 R 17 R 18 W, P, K, B, and J are as defined in this article.

[0050] The present invention also provides methods for preparing compounds of formulas I-1, I-2, I-3, I-4, I-5, II-1, II-2, II-3, and III-1, pharmaceutically acceptable salts thereof, or solvates thereof.

[0051] On the other hand, the present invention provides a pharmaceutical composition comprising the compounds described herein, as well as pharmaceutically acceptable salts and excipients thereof.

[0052] The present invention also provides methods for preparing compounds of formulas I-1, I-2, I-3, I-4, I-5, II-1, II-2, II-3, and III-1, their pharmaceutically acceptable salts, or solvates thereof.

[0053] in,

[0054] (1) The compounds of formula I-2 or I-4 are prepared according to the following steps:

[0055] Will The compound is reacted with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base, further reacted in the presence of a suitable base and a nucleophile, deprotected under H2 and Pd / C catalysis, reacted with a suitable bromide in the presence of a suitable base and Pd catalysis, deBoc was removed in the presence of TFA, and reacted with a suitable acylation reagent in the presence of a suitable base to give a compound of formula I-2 or I-4.

[0056] (2) The compounds of formula I-1, formula I-3 or formula I-5 are prepared according to the following steps;

[0057] Will The reaction is carried out with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base, further with a suitable base and a nucleophile, then with a suitable boron compound, with the Boc removed in the presence of TFA, and with a suitable acylation agent in the presence of a suitable base to give a compound of formula I-1, I-3 or I-5.

[0058] (3) The compound of formula II-2 is prepared according to the following steps;

[0059] Will The compound is reacted with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base, then in the presence of an oxidant, further in basic conditions, deprotected under H2 and Pd / C catalysis, reacted with a suitable bromide in the presence of a suitable base and under Pd catalysis, then deBoc is removed in the presence of TFA, and reacted with a suitable acylation reagent in the presence of a suitable base to give the compound of formula II-2.

[0060] (4) The compounds of formula II-1 or II-3 are prepared according to the following steps;

[0061] Will The reaction is carried out with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base, further under basic conditions, followed by reaction with a suitable boron compound, removal of the Boc in the presence of TFA, and reaction with a suitable acylation agent in the presence of a suitable base to give a compound of formula II-1 or II-3.

[0062] (5) The compound of formula III-1 is prepared according to the following steps:

[0063] Will The compound is reacted with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base, further reacted in the presence of a suitable base and a nucleophile, deprotected under H2 and Pd / C catalysis, reacted with a suitable bromide in the presence of a suitable base and Pd catalysis, deBoc is removed in the presence of TFA, and reacted with a suitable acylation reagent in the presence of a suitable base to give a compound of formula I-2 or I-4.

[0064] The present invention also provides pharmaceutical compositions comprising the compounds described herein, as well as pharmaceutically acceptable salts and excipients thereof.

[0065] The present invention also provides the use of the said compound, its isomers, its pharmaceutically acceptable salts or their deuterated derivatives, or the pharmaceutical composition thereof in the preparation of a medicament for treating tumors associated with KRAS G12C mutations.

[0066] The tumors associated with the KRAS G12C mutation include those located in the following sites: lung, colorectal, pancreas, brain, head and neck, liver, stomach, esophagus, breast, cervix, ovary, endometrium, larynx, oral cavity, prostate, thyroid, and soft tissue.

[0067] Preferably, the tumors include: lung cancer (non-small cell lung cancer, small cell lung cancer, squamous cell carcinoma), pancreatic cancer (ductal adenocarcinoma, islet tumor), gastric cancer, esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma), small intestine (adenocarcinoma), kidney (adenocarcinoma, nephroblastoma), bladder and urethra (squamous cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testicular cancer (seminomatous tumor), hepatocellular carcinoma, bile duct carcinoma, astrocytoma, glioblastoma, retinoblastoma, endometrial cancer, cervical cancer, ovarian cancer, sarcoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, osteosarcoma, fibrosarcoma, chondrosarcoma, multiple myeloma, myelodysplastic syndrome, Hodgkin's disease, non-Hodgkin's lymphoma, malignant melanoma, and hemangioma.

[0068] The present invention also provides a composition comprising the compound described herein, its isomers, pharmaceutically acceptable salts thereof, or their deuterated derivatives, as well as other antitumor drugs.

[0069] The present invention also provides the use of the compound, its isomers, its pharmaceutically acceptable salts or their deuterated derivatives, or the pharmaceutical composition thereof, or the composition thereof, in combination with a therapy such as radiotherapy or chemotherapy.

[0070] The present invention also provides the use of the compound, its isomers, its pharmaceutically acceptable salts or their deuterated derivatives, or the pharmaceutical composition thereof, or the composition thereof, in inhibiting the KRAS G12C mutant protein.

[0071] The present invention also provides the use of the compound, its isomers, its pharmaceutically acceptable salts or their deuterated derivatives, or the pharmaceutical composition thereof, or the composition thereof, in the preparation of a medicament for inhibiting the KRAS G12C mutant protein.

[0072] The present invention also provides a method for inhibiting the KRAS G12C mutant protein, comprising administering to a mammal in need an effective amount of the compound of the present invention, its isomer, its pharmaceutically acceptable salt or its deuterated form, or the pharmaceutical composition thereof, or the composition thereof.

[0073] The present invention also provides a method for treating KRAS G12C mutation-associated tumors, comprising administering to a mammal in need an effective amount of the compound, its isomers, its pharmaceutically acceptable salts or their deuterated derivatives, or the pharmaceutical composition, or the composition thereof, wherein the KRAS G12C mutation-associated tumors include tumor sites of occurrence such as: lung, colorectal, pancreas, brain, head and neck, liver, stomach, esophagus, breast, cervix, ovary, endometrium, larynx, oral cavity, prostate, thyroid, and soft tissue;

[0074] Preferably, the tumors include: lung cancer (non-small cell lung cancer, small cell lung cancer, squamous cell carcinoma), pancreatic cancer (ductal adenocarcinoma, islet tumor), gastric cancer, esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma), small intestine (adenocarcinoma), kidney (adenocarcinoma, nephroblastoma), bladder and urethra (squamous cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testicular cancer (seminomatous tumor), hepatocellular carcinoma, bile duct carcinoma, astrocytoma, glioblastoma, retinoblastoma, endometrial cancer, cervical cancer, ovarian cancer, sarcoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, osteosarcoma, fibrosarcoma, chondrosarcoma, multiple myeloma, myelodysplastic syndrome, Hodgkin's disease, non-Hodgkin's lymphoma, malignant melanoma, and hemangioma.

[0075] Preferably, the present invention also provides the use of compounds of formula I-1, formula I-2, formula I-3, formula I-4, formula I-5, formula II-1, formula II-2, formula II-3, formula III-1, or pharmaceutically acceptable salts thereof or solvates thereof in the preparation of medicaments for treating KRAS G12C-related cancers and diseases. Detailed Implementation

[0076] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention.

[0077] This invention relates to compounds that irreversibly inhibit the KRAS G12C mutant protein, pharmaceutical compositions comprising said compounds, and methods of application thereof.

[0078] definition

[0079] Unless otherwise defined, the terms and techniques used herein have the same meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0080] The “KRAS G12C” mentioned in this article refers to the KRAS protein mutation caused by the substitution of glycine at position 12 with cysteine.

[0081] The “KRAS G12C inhibitor” described herein refers to the compounds shown in Formulas I and II of this invention; these compounds bind irreversibly to KRAS G12C by forming a covalent bond with the cysteine ​​residue at position 12 of the KRAS mutant protein, resulting in the inhibition of KRAS G12C.

[0082] The “KRAS G12C-related cancers or diseases” mentioned in this article refer to cancers or diseases that are associated with or caused by KRAS G12C mutations.

[0083] The term "alkyl" refers to a straight-chain or branched aliphatic group containing 1 to 10, preferably 1 to 8, more preferably 1 to 6, and even more preferably 1 to 4 carbon atoms, optionally substituted by one or more substituents; exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, and hexyl.

[0084] The term "alkylene" refers to a group in which an alkyl group as defined above is located between and connects two chemical groups; exemplary alkylene groups include, but are not limited to, methylene, ethylene, propylene, and butylene.

[0085] The term "haloalkyl" refers to an alkyl chain in which one or more hydrogen atoms are replaced by halogens; exemplary haloalkyls include, but are not limited to, fluoromethyl, difluoromethyl, and trifluoromethyl.

[0086] The term "hydroxyalkyl" refers to -alkyl-OH.

[0087] The term "haloalkoxy" refers to -O-haloalkyl.

[0088] The term "alkoxy" refers to -O-(C1-C5) alkyl.

[0089] The term "cycloalkyl" refers to a saturated or partially saturated cyclic hydrocarbon group having 3 to 12, preferably 3 to 10, more preferably 3 to 6 carbons, which may optionally be substituted. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0090] The term "heteroalkyl" refers to an alkyl group defined above in which one or more carbon atoms are replaced by O, S, and N atoms.

[0091] The term "amino" refers to -NH2.

[0092] The term "heterocyclic group" or "heterocyclic group" refers to a 3-12 membered ring, preferably a 3-10 membered ring, and more preferably a 3-6 membered ring, in which one or more nitrogen, oxygen, or sulfur atoms are included in addition to carbon atoms. The "heterocyclic group" or "heterocyclic group" can be monocyclic, bicyclic, spirocyclic, or bridged ring; the aforementioned "heterocyclic group" or "heterocyclic group" may optionally be substituted at one or more carbon or nitrogen positions; exemplary "heterocyclic groups" or "heterocyclic groups" include, but are not limited to, piperazinyl, piperidinyl, epoxy, aziridinyl, furanyl, pyrrolyl, hexahydropyridinyl, tetrahydropyrrolyl, tetrahydrofuranyl, morpholinyl, pyrrolidoneyl, thiazolyl, oxazolyl, hexahydropiperidinyl, acridineyl, thiophene, tetrahydrothiophene, piperidinyl, imidazole, indole, pyridinyl, pyrimidinyl, pyrazinyl, decahydroquinolinyl, piperidinoneyl, etc.

[0093] The term "heterocyclic alkyl" refers to a group that is attached to the alkyl group linked to the heterocyclic group and then to the rest of the molecule. Exemplary "heterocyclic alkyl" includes, but is not limited to, azirrocyclobutane, oxacyclopropane, thiazolyl, pyrroleyl, imidazolyl, azirbicyclohexane, azirbicycloheptane, etc.

[0094] The term "aryl" refers to an aromatic ring containing 6-20, preferably 6-14, more preferably 6-12, and even more preferably 6-10 carbon atoms, which may optionally be substituted; exemplary aryl groups include, but are not limited to, phenyl, naphthyl, anthracene, and fluorenyl.

[0095] The term "aralkyl" refers to an aryl group covalently linked to an alkyl group, which may optionally be substituted; exemplary aralkyl groups include, but are not limited to (C6-C10)aryl (C1-C5)alkyl, preferably benzyl and naphthylethyl.

[0096] The term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic group composed of 5-14, preferably 6-14, more preferably 6-10 ring atoms; these monocyclic, bicyclic, or tricyclic groups composed of "heteroaryl" share 6, 10, or 14 π electrons in the cyclic array of the aromatic heterocyclic compound; the atoms constituting the ring contain one or more heteroatoms selected from N, O, and S in addition to carbon atoms; exemplary "heteroaryl" or "heteroaryl group" includes, but is not limited to, pyranyl, pyridyl, pyrimidinyl, and pyridazinyl. Pyrazinyl, oxazinyl, benzofuranyl, indoleyl, purinyl, inzolyl, benzopyranyl, benzopyranoneyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinazinyl, pteridinyl, naphthidyl, phthalazinyl, quinoxalinyl, quinazolinyl, carbazoleyl, phenazinyl, acridineyl, benzimidazolyl, pyrrolopyridyl, pyridopyrimidinyl, thienopyrroleyl, imidazothiazolyl, acridineyl, acridineyl, benzothiopheneyl, benzothiazolyl, benzooxazolyl, benzotriazolyl, benzotetrazoleyl, benzimidazolyl oxazolyl, benzisothiazolyl, benzimidazolinyl, carbolinyl, chromanyl, isochromyl, isoydinyl, chromenyl, furanyl, imidazolinyl, imidazolyl, isoydinolyl, isothiazolyl, 1H-indazoleyl, indoleyl, inazinyl, trihydroindoleyl, isobenzofuranyl, isoxazolyl, naphridinyl, diazolyl, methylenedioxyphenyl, octahydroisoquinolinyl, tetrahydroisoquinolinyl, isodihydroindoleyl, oxazolyl, phenanthridineyl, phenothiazinyl, pyrazolylyl, tetrahydroquinazinyl, quininecycloyl, tetrazolyl Pteridyl, phenazinyl, pyrazolyl, pyrazolinyl, pyridinium-imidazolyl, pyridothiazolyl, pyrrololinyl, pyrroleyl, dihydropyrroleyl, dihydroindolyl, pyridothiazolyl, oxazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, thiazolyl, thiophenyl, thiophenothiazolyl, thiophenooxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl and thiopheninium-imidazolyl, etc.

[0097] The term "heteroarylalkyl" refers to a group that is connected to other parts of a molecule via an alkyl group linked to a heteroaryl group, wherein each of the heteroarylalkyl groups may be independently and optionally substituted. Exemplary heteroarylalkyl groups include, but are not limited to, isoquinolinylmethyl, tetrahydroisoquinolinylmethyl, quinolinylmethyl, tetrahydroquinolinylmethyl, quinolinylethyl, tetrahydroquinolinylethyl, quinazolinylmethyl, pyrrolithylmethyl, pyrrolithylethyl, pyridylmethyl, pyrimidinylmethyl, indolylmethyl, isoindolylmethyl, imidazolylmethyl, imidazolylcyclopropyl, pyrrolithylisopropyl, benzimidazolylmethyl, thiazolylmethyl, and pyridylethyl.

[0098] In the above definition, "one or more" can refer to one or two, one or three, one or four, one or five, one or six, one or eight, one or ten, or one or more.

[0099] compound

[0100] In a first aspect, the present invention provides compounds of formula I, II, or III, isomers thereof, pharmaceutically acceptable salts thereof, or deuterated derivatives thereof:

[0101]

[0102] in:

[0103] W is a 4-12 element saturated or partially saturated monocyclic, double-ring, bridged, or helical ring, wherein the saturated or partially saturated monocyclic, double-ring, bridged, or helical ring is optionally replaced by one or more R2.

[0104] P and K are nitrogen or carbon, preferably, P and K are both nitrogen or at least one of P and K is nitrogen;

[0105] B is either nonexistent or -NH-; when B is nonexistent, P is directly connected to R1;

[0106] R1 is

[0107]

[0108] E is CR7 or NR8;

[0109] G is CR9 or NR 10 ;

[0110] Q is CR 11 NR 12 N or C = O;

[0111] Is it a single bond or a double bond?

[0112] R2 is a C1-C3 alkyl, C2-C4 alkynyl, heteroalkyl, cyano or oxo, wherein the C1-C3 alkyl may be optionally substituted with halogen, cyano or heteroaryl.

[0113] R3 is absent, hydrogen, halogen, CF3, hydroxyalkyl, or C1-C3 alkyl;

[0114] R4 is absent, hydrogen, C1-C3 alkyl, alkylaminoalkyl, heterocyclic, aminoalkyl, halogen, amide, nitrile, hydroxyalkyl, CF3, CF2, methoxy, trifluoromethyl, amino or alkenyl;

[0115] R5 is absent, hydrogen, alkyl, or hydroxyalkyl;

[0116] R6 is absent, hydrogen, or a C1-C3 alkyl group;

[0117] R7 is hydrogen, C1-C4 alkyl, cycloalkyl, amino, cyano, alkynyl, halogen, oxo, aminoalkyl, alkylaminoalkyl, or haloalkyl.

[0118] R8 is hydrogen, C1-C4 alkyl, cycloalkyl, alkynyl, or -CONH2;

[0119] R9 is a cycloalkyl, heterocyclic, aryl, aralkyl, or heteroaryl group; the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups may be optionally combined to form monocyclic, bicyclic, or tricyclic groups; each of the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups may be represented by one or more R9 groups. 13 replace;

[0120] R 10 It is a cycloalkyl, heterocyclic, aryl, aralkyl, or heteroaryl group; the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups can be optionally combined to form monocyclic, bicyclic, or tricyclic groups; each of the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups can be represented by one or more R groups. 15 replace;

[0121] R 11 It is absent, hydrogen, C1-C4 alkyl, cycloalkyl, amino, or halogen;

[0122] R 12 It is hydrogen, C1-C4 alkyl, cycloalkyl, or alkynyl;

[0123] R 13 It is a halogen, hydroxyl, C1-C6 alkyl, cycloalkyl, alkoxy, haloalkyl, amino, cyano, heteroalkyl, hydroxyalkyl, oxoalkyl, or thioalkyl;

[0124] R 14 It is hydrogen, alkyl, aminoalkyl, alkylaminoalkyl, haloalkyl, hydroxyalkyl, dihydroxyalkyl, 4-6 membered heterocyclic, 4-6 membered heterocyclic alkyl, 5-6 membered aryl, 5-6 membered heteroaryl or heteroarylalkyl, wherein the heterocyclic, heterocyclic alkyl, aryl, heteroaryl or heteroarylalkyl may be optionally surrounded by one or more identical or different R 15 replace;

[0125] R 15 It is a halogen, hydroxyl, hydrogen, C1-C6 alkyl, cycloalkyl, alkoxy, haloalkyl, amino, cyano, heteroalkyl, oxoalkyl, aralkyl, or acyl.

[0126] R 17 It is a cycloalkyl, heterocyclic, aryl, aralkyl, or heteroaryl group; each of the cycloalkyl, heterocyclic, aryl, aralkyl, and heteroaryl groups may optionally be converted by one or more R groups. 16 replace;

[0127] R16 It is hydrogen, C1-C4 alkyl, amino, cyano, alkynyl, halogen, oxo, aminoalkyl, or haloalkyl;

[0128] R 18 It is hydrogen, C1-C4 alkyl, cycloalkyl, amino, halogen, or trifluoromethyl;

[0129] J is either O or S.

[0130] In some implementations, W is independently:

[0131]

[0132] In some implementations,

[0133] yes

[0134] In some implementations, R1 is R3 and R4 are as defined in this paper.

[0135] In some implementations, R1 is R5 is as defined in this article.

[0136] In some implementations, R1 is R6 is as defined in this article.

[0137] Preferably, R1 is independently:

[0138]

[0139] In some embodiments, the invention also includes compounds of formula I-1, I-2, I-3, I-4, I-5, II-1, II-2, II-3, and III-1, their isomers, pharmaceutically acceptable salts thereof, or deuterated derivatives thereof:

[0140]

[0141] Among them, R1, R2, R7, R8, R9, R 10 R 11 R 12 R 14 R 16 R 17 R 18 W, P, K, B, and J are as defined in this document.

[0142] In a preferred embodiment, the present invention includes compounds of formulas I-1, I-2, I-3, I-4, I-5, II-1, II-2, II-3, and III-1, their pharmaceutically acceptable salts, and pharmaceutical compositions thereof. The present invention also provides compounds represented by the following formulas, their isomers, their pharmaceutically acceptable salts, or their deuterated derivatives:

[0143]

[0144]

[0145]

[0146]

[0147]

[0148] In the specific compounds above, R and S represent stereoconfigurations, respectively.

[0149] Pharmaceutical Composition

[0150] The pharmaceutical composition of the present invention refers to a pharmaceutical composition that is convenient for administration, prepared by means of the compounds of the present invention (including racemates, enantiomers, stereoisomers, deuterated derivatives) or their pharmaceutically acceptable salts, hydrates, solvates, prodrugs and their pharmaceutically acceptable carriers, excipients or adjuvants.

[0151] Preferably, the pharmaceutical composition of the present invention comprises the KRAS G12C inhibitory compound of the present invention and a pharmaceutically acceptable excipient or carrier. The administration routes of the compounds and pharmaceutical compositions of the present invention can be: 1) oral: e.g., tablets, capsules, etc.; 2) injection: e.g., intravenous injection, subcutaneous injection, intramuscular injection, ocular injection, etc.; 3) rectal: e.g., suppositories, gels, etc.; (4) nasal inhalation: e.g., sprays, aerosols, etc.; administration can also be via drug release systems such as liposomes, sustained-release technology, and controlled-release technology. The preferred methods are oral or injectable administration.

[0152] The term "pharmaceutically acceptable salt" refers to a salt that maintains the biological activity of the compounds of the present invention without exhibiting undesirable toxicological effects. Examples of pharmaceutically acceptable salts of the compounds of the present invention include, but are not limited to, acid addition salts formed with inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, and hydrobromic acid, as well as salts formed with organic acids such as acetic acid, malic acid, tartaric acid, oxalic acid, succinic acid, benzoic acid, tannic acid, alginic acid, and polyglutamic acid; the compounds of the present invention can also be administered as pharmaceutically acceptable quaternary ammonium salts.

[0153] In addition to containing the compounds of the present invention or their pharmaceutically acceptable salts, the pharmaceutical compositions described herein may also contain buffers, diluents, fillers, stabilizers, solubilizers, and other excipients disclosed in the art.

[0154] The various dosage forms of the pharmaceutical compositions composed of the compounds of the present invention are prepared using methods commonly used in the pharmaceutical industry, including but not limited to mixing, dissolving, granulating, grinding, emulsifying, sugar coating, freeze drying, etc.

[0155] The "pharmaceutically acceptable carrier" of this invention refers to the inactive ingredients in the pharmaceutical composition, including but not limited to: calcium phosphate, calcium carbonate, various sugars such as lactose or mannitol, starch, magnesium stearate, cellulose, magnesium carbonate, acrylic polymers, methacrylic polymers, gels, ethylene glycol, castor oil, sesame oil, corn oil, peanut oil, etc.

[0156] The pharmaceutical compositions containing compounds that inhibit KRAS G12C and the methods of application provided by this invention can be used to treat various tumors associated with KRAS G12C mutations, including but not limited to the following tumor sites: lung, colorectal, pancreas, brain, head and neck, liver, stomach, esophagus, breast, cervix, ovary, endometrium, larynx, oral cavity, prostate, thyroid, and soft tissue. More specifically, the compounds or pharmaceutical compositions of the present invention can be used to treat: lung cancer (non-small cell lung cancer, small cell lung cancer, squamous cell carcinoma), pancreatic cancer (ductal adenocarcinoma, islet tumor), gastric cancer, esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma), small intestine (adenocarcinoma), kidney (adenocarcinoma, nephroblastoma), bladder and urethra (squamous cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testicular cancer (seminomatous seminoma), hepatocellular carcinoma, bile duct carcinoma, astrocytoma, glioblastoma, retinoblastoma, endometrial cancer, cervical cancer, ovarian cancer, sarcoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, osteosarcoma, fibrosarcoma, chondrosarcoma, multiple myeloma, myelodysplastic syndrome, Hodgkin's disease, non-Hodgkin's lymphoma, malignant melanoma, and hemangioma.

[0157] The compounds and pharmaceutically acceptable salts thereof described in this invention, as well as pharmaceutical compositions containing such compounds or their salts, may be used in combination with other antitumor drugs or therapies such as radiotherapy or chemotherapy.

[0158] Reaction schemes and examples

[0159] The present invention will be described in detail below through embodiments, but these embodiments are only for describing implementation schemes of the present invention and are not intended to limit the scope of the present invention. The compounds of the present invention can be prepared by various synthetic methods well known to those skilled in the art, including but not limited to the methods used in the embodiments of the present invention and alternative methods well known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention. Various changes and substitutions made by those skilled in the art to the technical solutions of the present invention based on the design concept of the present invention are all within the protection scope of the present invention.

[0160] Reaction scheme

[0161] When G is nitrogen, the compounds of formula I-2 and I-4 are synthesized according to reaction IA;

[0162] When G is carbon, the compounds of formula I-1, I-3 and I-5 are synthesized according to reaction scheme IB;

[0163] When G is nitrogen, the compound of formula II-2 is synthesized according to reaction scheme II-C;

[0164] When G is carbon, the compounds of formulas II-1 and II-3 are synthesized according to reaction scheme II-D;

[0165] When G is nitrogen, the compound of formula III-1 is synthesized according to reaction scheme III-E.

[0166] Solution IA:

[0167]

[0168] As shown in scheme IA, where W, P, K, B, E, Q, J, R1, R2, R 10 R 14 As defined herein, intermediate (1) reacts with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base (e.g., PIPEA) to form (2), which reacts with a suitable base (e.g., NaH) and a nucleophile (e.g., a substituted alcohol or amine) to form (3), which is deprotected by H2 and Pd / C catalysis to form (4), which reacts with a suitable bromide in the presence of a suitable base (SeCO3) and Pd catalysis to form (5), which is deBoc-protected by TFA to form (6), which is reacted by a suitable acylation agent (e.g., acryloyl chloride) in the presence of a suitable base (e.g., TEA) to give compound (I).

[0169] Option IB:

[0170]

[0171] As shown in scheme IB, W, P, K, B, E, Q, R1, R2, R9, R 14 As defined herein, intermediate (1) reacts with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base (e.g., PIPEA) to form (2), which then reacts with a suitable base (e.g., NaH) and a nucleophile (e.g., a substituted alcohol or amine) to form (3), which then reacts with a suitable boron compound to form (4), which then undergoes debonding in the presence of TFA to form (5), which then reacts with a suitable acylation agent (e.g., acryloyl chloride) in the presence of a suitable base (e.g., TEA) to give compound (I).

[0172] Scheme II-C

[0173]

[0174] As shown in Scheme II-C, W, P, K, B, E, Q, R1, R2, R 10 R 17 As defined herein, intermediate (1) reacts with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base (e.g., PIPEA) to form (2), structure (2) in the presence of an oxidizing agent to form (3), structure (3) in basic conditions to form (4), structure (4) in basic conditions to form (5), structure (5) in H2 and Pd / C catalysis to form (6), structure (6) in a suitable base (SeCO3) in Pd catalysis to form a suitable bromide to form (7), structure (7) in the presence of TFA to form (8), structure (8) in the presence of a suitable base (e.g., TEA) to form a suitable acylation agent (e.g., acryloyl chloride) to give compound (II).

[0175] Solution II-D

[0176]

[0177] As shown in Scheme II-D, W, P, K, B, E, Q, R1, R2, R9, R 17 As defined herein, intermediate (1) reacts with a Boc-protected amine or a Boc-protected boron-containing compound in the presence of a suitable base (e.g., PIPEA) to form (2), structure (2) reacts under basic conditions to form (3), structure (3) reacts under basic conditions to form (4), structure (4) reacts with a suitable boron compound to form (5), structure (5) loses its Boc in the presence of TFA to form (6), and structure (6) reacts with a suitable acylation agent (e.g., acryloyl chloride) in the presence of a suitable base (e.g., TEA) to give compound (II).

[0178] Option III-E:

[0179] Compound (III) was prepared according to the same method as in scheme IA. The difference lies in the starting material.

[0180] Example 1

[0181]

[0182] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-5-methyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinoline-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in the reaction procedure of Example 1; LCMS (ESI) m / z: 546.3 [M+H] + .

[0183] Synthetic route of Example 1:

[0184]

[0185] Step 1: A mixed solution of compound 1 (1.09 mL, 8.76 mmol) and benzylamine (1.05 mL, 9.64 mmol) was reacted at 100 °C for 18 h. The reaction mixture was concentrated under reduced pressure and purified to give compound 2 (1.1 g, 56%), MS (M+H). + =222.

[0186] Step 2: A 21% sodium ethoxide solution (21:79, sodium ethoxide: ethanol, 2 mL) was added dropwise to a mixture of compound 2 (1.1 g, 4.97 mmol) and diethyl oxalate (0.75 mL, 5.52 mmol). The reaction mixture was stirred at room temperature for 1 hour. The mixture was monitored by TLC, concentrated, and then water (approximately 10 mL) was added. After stirring for 5 minutes, the pH was adjusted to 1 with 1 M HCl, and the mixture was filtered. The solid was collected, dried under vacuum overnight, and then crystallized from ethanol to give compound 3 (0.8 g, 58%), MS (M+H). + =276.

[0187] Step 3: Compound 3 (2 g, 7.26 mmol) and ammonium formate (0.925 g, 14.67 mmol) were added to ethanol (10 mL), and the mixture was stirred at 78 °C for 24 h. After the reaction was complete, the reaction mixture was concentrated, the resulting solid was washed with water, filtered, and dried. Recrystallization from ethanol gave compound 4 (1.5 g, 75%) MS(M+H). + =275.

[0188] Step 4: Guanidine hydrochloride (2.77 g) was dissolved in ethanol (30 mL), then a 21% sodium ethoxide solution in ethanol (21:79, sodium ethoxide: ethanol, 13 mL) was added, followed by compound 4 (1.5 g). The reaction mixture was then stirred at 78 °C for 3 days. The reaction mixture was concentrated, and then water (30 mL) was added. The mixture was stirred until all solids were dissolved, and then glacial acetic acid was added dropwise until pH = 6. The mixture was filtered, the solids were washed with ethanol and dried, and recrystallized from DMF to give compound 5 (0.5 g, 33%) MS(M+H). + =271.

[0189] Step 5: Mix compound 5 (2.5 g) and water (150 mL), and heat the mixture to 100 °C. Slowly add concentrated hydrochloric acid until the solution becomes clear. Continue adding 8 mL of concentrated hydrochloric acid, cool the mixture to 90 °C, and add sodium nitrite aqueous solution (1.9 g dissolved in 20 mL of water) dropwise. Continue stirring at 90 °C for 30 min, then filter while hot to obtain compound 6 (0.9 g, 35%) MS (M+H). + =272.

[0190] Step 6: Add compound 6 (0.5 g), POCl3 (8.50 mL), and N-diethylaniline (0.53 mL) to the flask. Stir the mixture overnight at 106 °C. Pour the reaction mixture onto ice and extract three times with CH2Cl2. Combine the organic phases, dry them over anhydrous sodium sulfate, filter, and concentrate. Purify by silica gel chromatography (0-50% EtOAc petroleum ether solution) to give compound 7 (0.35 g, 61%). MS (M+H) + =309.

[0191] Step 7: Compound 7 (0.5 g, 1.62 mmol), piperazine-1-carboxylic acid tert-butyl ester (0.32 g, 1.7 mmol), and DIPEA (4.06 mmol) were dissolved in DMSO and reacted at 55 °C for 10 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and subjected to column chromatography (0-50% EA in petroleum ether solution) to give compound 8 (0.65 g, 87%). MS (M+H) + =459.

[0192] Step 8: N-methyl-L-prolyl (0.75 g, 6.55 mmol) and THF (20 ml) were added to a round-bottom flask. The mixture was placed in an ice bath, and NaH (0.39 g, 9.83 mmol) was added in portions with stirring. The mixture was reacted under nitrogen protection in an ice bath for 40 min. Compound 8 (1.5 g, 3.28 mmol) was then added, and the reaction was carried out overnight at 70 °C. After the reaction was complete, a saturated ammonium chloride solution was added, and the aqueous layer was extracted with EA (30 ml × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by silica gel column chromatography to give compound 9 (1.08 g, 61%). MS (M+H) + =537.

[0193] Step 9: Compound 9 (1.8 g, 2.79 mmol) was dissolved in methanol (20 ml), and Pd(OH)₂ / C (500 mg, 10%) was added. The mixture was purged three times with hydrogen gas, and reacted at 40 °C for 12 h under hydrogen protection. After the reaction was complete, the mixture was filtered, the filtrate was concentrated, and purified by silica gel chromatography to obtain compound 10 (0.7 g, 56%). MS (M+H) + =447.

[0194] Step 10: Compound 10 (0.5 g, 1.12 mmol) and 5-bromo-2-methyltetrahydroisoquinoline (0.379 g, 1.68 mmol) were dissolved in 8 mL of anhydrous toluene. Under nitrogen protection, Pd₂(dba)₃ (153 mg), RuPhos (104 mg), and Cs₂CO₃ (729 mg) were added sequentially. The reaction was carried out at 110 °C for 12 h under nitrogen protection. After the reaction was complete, EA and water were added, and the mixture was extracted twice with EA (30 mL × 2). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography to give compound 11 (0.34 g, 51%). MS (M+H) + =592.

[0195] Step 11: Compound 11 (0.5 g) was dissolved in 2.5 ml of DCM, and TFA (2 ml) was slowly added dropwise under ice bath conditions. The reaction mixture was continued to react under ice bath conditions for 1 h, and the reaction was monitored by TLC. The reaction solution was concentrated to obtain the crude product of compound 12.

[0196] Step 12: Dissolve the crude product of compound 12 from the previous step in 4 ml of DCM, slowly add DIPEA (900 μL), and add acryloyl chloride (65 μL) dropwise under ice bath conditions. Continue the reaction for 10 min, and monitor the reaction by TLC. After the reaction is complete, add saturated NaHCO3, extract with EA (30 ml × 2), combine the organic phases, dry with anhydrous Na2SO4, filter, concentrate, and purify by preparative HPLC (ACN-H2O, C18 column) to obtain Example 1, LCMS (ESI) m / z 546.3.

[0197] Example 2

[0198]

[0199] The synthesis of (S)-1-(4-(5-isopropyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is shown in the reaction procedure of Example 2; LCMS (ESI) m / z: 560.3 [M+H] + .

[0200] Synthetic route of Example 2:

[0201]

[0202] Step 1: Compound 1 (1.02 mL, 6.32 mmol) and benzylamine (690 μL, 6.32 mmol) were dissolved in anhydrous toluene (8 mL) and refluxed at 110 °C (Dean-Stark separator) for 3 h under nitrogen protection. The solvent was evaporated to dryness, and the residue was dissolved in 10 mL of glacial acetic acid. NaCNBH3 (1.8 g, 28.45 mmol) was added in portions, and the reaction mixture was stirred at room temperature for 2 h. The residue was extracted with diethyl ether, the organic layer was washed with 1N NaOH and saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give compound 1 (1.2 g, 76%). MS (M+H) + =250.

[0203] Step 2: Compound 1 (1.5 g, 6.02 mmol), methyl bromoacetate (677 μL, 7.16 mmol), and K₂CO₃ (1.66 g, 12.3 mmol) were dissolved in acetonitrile (20 mL), stirred overnight at room temperature, and the reaction solution was filtered through diatomaceous earth. The concentrated compound 2 (1.6 g, 82%) was analyzed by MS (M+H). + =322.

[0204] Step 3: Compound 2 (1.6 g) was dissolved in 10 ml of toluene. The mixture was kept in an ice bath, and 6 ml of 1 N potassium tert-butoxide tetrahydrofuran solution was slowly added dropwise. The mixture was stirred overnight at room temperature. 10 ml of 1 N HCl was added, and stirring continued for 8 hours. The reaction was terminated by adding Na₂CO₃. The mixture was extracted with DCM (50 ml × 3), and the organic phases were combined, washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to obtain compound 3 (1.05 g, 72%). MS (M+H) + =290.

[0205] Step 4: Refer to step 4 of Example 1.

[0206] Step 5: Compound 5 (3g) was added to 30ml of phosphorus oxychloride and refluxed overnight. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution (100ml) was slowly added dropwise under ice bath, extracted with ethyl acetate, dried over anhydrous Na2SO4, filtered, and concentrated to obtain compound 6 (3.1g) by MS (M+H). + =323.

[0207] Steps 6 to 11, referring to Example 1, yielded Example 2 (200mg), LCMS (ESI) m / z 560.3.

[0208] Example 3

[0209]

[0210] The synthesis of (S)-1-(4-(7-methyl-6-(8-methylnaphthyl-1-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is shown in the synthetic reaction procedure of Example 3; LCMS (ESI) m / z: 525.2

[0211] [M+H] + .

[0212] Synthetic route of Example 3:

[0213]

[0214] Step 1: Compound 1 (1 g, 5.32 mmol) was dissolved in anhydrous THF (10 mL). NaH (320 mg, 60%, 7.98 mmol) was added in portions under ice bath conditions, and the reaction was continued for 50 min. Benzenesulfonyl chloride (1.22 g, 6.91 mmol) was then added, and the reaction was carried out at room temperature for 1.5 h. The reaction was monitored by TLC until completion. Saturated NH4Cl solution (50 mL) was slowly added, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to give target compound 2 (1.53 g, 87%). LC / MS (ESI): 328.

[0215] Step 2: Compound 2 (910 mg, 2.77 mmol) was dissolved in anhydrous THF (10 ml), cooled to -78 °C, and diisopropylaminolithium solution (5 ml, 2 M dissolved in THF / n-heptaneethylbenzene) was slowly added dropwise. The reaction was continued for 30 min, and 1,2-dibromotetrachloroethane (2.71 g, 8.32 mmol) was added. The mixture was stirred at -78 °C for 2 h. After the reaction was completed by TLC monitoring, saturated NH4Cl solution (20 ml) was added, and the mixture was extracted with EA (3×). The mixture was dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated to dryness under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 3 (650 mg, 57%), LC / MS (ESI): 408.

[0216] Step 3: Compound 3 (650 mg, 1.6 mmol) was dissolved in anhydrous THF (15 ml), potassium tert-butoxide (896 mg, 7.98 mmol) was added, and the mixture was left at room temperature overnight. The reaction was quenched by adding saturated NaHCO3, extracted with EA, dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 4 (240 mg, 56%). LC / MS (ESI): 268.

[0217] Step 4: Compound 4 (240 mg, 899 μmol) was dissolved in anhydrous THF (6 ml), and NaH (54 mg, 60%, 1.35 mmol) was added in portions under ice bath. The reaction was continued under ice bath for 30 min. Iodimethane (168 μL, 2.7 mmol) was added dropwise to the reaction mixture, and the mixture was left to stand overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was quenched with water, extracted with EA, dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated to dryness under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 5 (148 mg, 59%), LC / MS (ESI): 281.

[0218] Step 5: Compound 5 (351 mg, 1.25 mmol), tert-butyl piperazine-1-carboxylate (256 mg, 1.37 mmol), and DIPEA (620 μL, 3.75 mmol) were dissolved in DMSO and reacted at 60 °C for 3 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain compound 6 (320 mg, 59%). MS (M+H) + =432.

[0219] Step 6: N-methyl-L-prolyl (112 mg, 975 μmol) was dissolved in THF (4 ml), and NaH (60 mg, 60%, 1.47 mmol) was added in portions under ice bath conditions. The reaction was carried out under nitrogen protection for 40 min on ice. Compound 6 (210 mg, 487 μmol) was then added, and the reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the reaction solution was slowly added to a saturated ammonium chloride solution. The aqueous layer was extracted with EA (30 ml × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by silica gel column chromatography to obtain compound 7 (118 mg, 47%). MS (M+H) + =510.

[0220] Step 7: Compound 7 (118 mg, 232 μmol), 8-methylnaphthalene-1-boronic acid (51.7 mg, 278 μmol), potassium acetate (56 mg, 579 μmol), Pd(dppf)2Cl2 (17 mg, 23 μmol), dioxane (4 ml), and water (1 ml) were refluxed overnight under nitrogen protection. The reaction was monitored by TLC. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with EA. The combined organic phases were washed with saturated brine, dried over anhydrous Na2SO4, filtered, concentrated, and purified by silica gel column chromatography to obtain compound 8 (56 mg, 46%). LC / MS (ESI): 571.

[0221] Step 8: Compound 8 (56 mg) was dissolved in 1 ml of DCM, and TFA (1 ml) was slowly added dropwise under ice bath. The reaction was carried out under nitrogen protection for 1 h. The reaction was monitored by TLC until it was completed. The reaction solution was concentrated to obtain the crude product of compound 9.

[0222] Step 9: Dissolve the crude product of compound 9 from the previous step in 2 ml of DCM, slowly add DIPEA (40 μL), and add acryloyl chloride (10 μL) dropwise under ice bath. Monitor the reaction completion by TLC. Quench the system with saturated NaHCO3, extract with EA (20 ml × 2), combine the organic phases, dry with anhydrous Na2SO4, filter, concentrate, and purify by preparative HPLC to obtain Example 3, LCMS (ESI) m / z 525.2.

[0223] Example 4

[0224]

[0225] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(8-chloronaphthyl-1-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in the synthetic reaction procedure of Example 4; LCMS (ESI) m / z: 547.2 [M+H] + .

[0226] Synthetic route of Example 4:

[0227]

[0228] Step 1: A 21% sodium ethoxide solution (21:79, sodium ethoxide: ethanol, 1.26 mL) was added dropwise to a mixture of ethyl β-benzylaminopropionate (384 μL, 2.41 mmol) and diethyl oxalate (360 μL, 2.65 mmol). The reaction mixture was stirred at room temperature for 1 hour under nitrogen protection. After the reaction was complete, the reaction mixture was concentrated, and then water (approximately 5 mL) was added. The mixture was stirred for 5 minutes, then adjusted to pH 1 with 1 M HCl and filtered under vacuum. The solid was collected and dried under vacuum overnight, then crystallized from ethanol to give compound 2 (366 mg, 58%).

[0229] Step 2: Compound 2 (366 mg, 1.4 mmol) and ammonium formate (176 mg, 2.81 mmol) were added to ethanol (2 mL), and the mixture was stirred at 78 °C for 24 h. After the reaction was complete, the mixture was evaporated to dryness, and the residue in ethanol was crystallized to give compound 3 (364 mg). MS (M+H) + =261.

[0230] Step 3: Guanidine hydrochloride (664 mg, 6.9 mmol) was dissolved in anhydrous ethanol (1 mL), then a 21% sodium ethoxide ethanol solution (21:79, sodium ethoxide: ethanol, 3 mL) was added, followed by compound 3 (360 mg, 1.38 mmol). The reaction mixture was stirred at 78 °C for 3 days. The reaction mixture was concentrated, and then water (10 mL) was added. Acetic acid was added dropwise with stirring until pH = 6. The mixture was filtered, ground in DMF, to give compound 4 (193 mg, 54%).

[0231] Step 4: Mix compound 4 (193 mg / L) with water (10 mL), heat to 100 °C, and slowly add concentrated hydrochloric acid until the sample dissolves. Cool to 90 °C, and slowly add dropwise a solution of sodium nitrite (155 mg) in water (1.5 mL). Stir at 90 °C for 1 hour, then filter while hot to obtain solid compound 5 (82 mg, 42%).

[0232] Step 5: Add compound 5 (80 mg), POCl3 (1.2 ml), and N-diethylaniline (74 μl) to the flask. Stir overnight at 106 °C. Slowly pour the reaction mixture into ice water and extract three times (30 ml × 3) with CH2Cl2. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate. The residue obtained is purified by silica gel chromatography to give compound 6 (40 mg, 43%).

[0233] Steps 6 to 11: Referring to Example 1, the sample was purified by HPLC to obtain Example 4, with an LCMS (ESI) m / z of 547.2.

[0234] Example 5

[0235]

[0236] The synthesis of (S)-1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is shown in the synthetic reaction procedure of Example 5;

[0237] LCMS(ESI) m / z: 518.3 [M+H] + .

[0238] Synthetic route of Example 5:

[0239]

[0240] Step 1: Compound 1 (2.37 g, 9.58 mmol) and urea (3.26 g, 54.29 mmol) were dissolved in 11 mL of anhydrous ethanol. A 21% sodium ethoxide ethanol solution (21:79, sodium ethoxide: ethanol, 1.4 mL) was slowly added dropwise and refluxed at 80 °C overnight. The ethanol was evaporated under reduced pressure, 20 mL of water was added, and the mixture was stirred for 30 min. The mixture was extracted with DCM (60 mL × 2), and the aqueous phase was collected. The pH was adjusted to approximately 4 with 1 N HCl, and the solid was collected by filtration and dried to obtain compound 2 (537 mg, 23%).

[0241] Step 2: Compound 2 (200 mg, 822 μmol) was added to phosphorus oxychloride (2 mL, 21.54 mmol), refluxed overnight at 110 °C under nitrogen protection. After the reaction was complete, the mixture was cooled, and 10 mL of saturated sodium bicarbonate was slowly added dropwise in an ice bath. The mixture was extracted with ethyl acetate (20 mL × 3), the organic phases were combined, dried over anhydrous Na₂SO₄, filtered, concentrated, dried over anhydrous Na₂SO₄, filtered, concentrated, and purified by silica gel column chromatography to obtain compound 3 (80 mg, 34%). MS (M+H) + =281.

[0242] Step 3: Compound 3 (300 mg, 1.07 mmol), piperazine-1-carboxylic acid tert-butyl ester (209 mg, 1.12 mmol), and DIPEA (350 μL, 2.14 mmol) were dissolved in DMSO and reacted at 60 °C for 3 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain compound 4 (280 mg, 60%). MS (M+H) + =431.

[0243] Step 4: N-methyl-L-prolyl (267 mg, 2.33 mmol) was dissolved in THF (8 ml), and NaH (139 mg, 60%, 3.49 mmol) was added in portions under ice bath conditions. The reaction was carried out under nitrogen protection for 40 min on ice. Compound 4 (500 mg, 1.16 mmol) was then added, and the reaction was carried out overnight at 70°C. After the reaction was completed, the reaction solution was slowly added to a saturated ammonium chloride solution. The aqueous layer was extracted with EA (30 ml × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by silica gel column chromatography to obtain compound 5 (360 mg, 60%). MS (M+H) + =509.

[0244] Step 5: Compound 5 (1 g, 1.97 mmol) was dissolved in methanol (10 ml), and Pd(OH)₂ / C (230 mg, 10%) was added. The mixture was reacted at 40 °C for 12 h under hydrogen protection. The mixture was filtered, the filtrate was concentrated, and purified by silica gel chromatography to obtain compound 6 (0.7 g, 85%). MS (M+H) + =419.

[0245] Step 6: Compound 6 (519 mg, 1.33 mmol) and 5-bromo-2-methyltetrahydroisoquinoline (389 mg, 1.72 mmol) were dissolved in 13 mL of anhydrous toluene. Under nitrogen protection, Pd₂(dba)₃ (121 mg, 132 μmol), RuPhos (123 mg, 265 μmol), and Cs₂CO₃ (1.3 g, 3.98 mmol) were added sequentially. The mixture was refluxed overnight at 110 °C under nitrogen protection. Extraction was performed twice with EA (30 mL × 2). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography to give compound 7 (0.48 g, 64%). MS (M+H) + =564.

[0246] Step 7: Compound 7 (0.38 g) was dissolved in 3 ml of DCM, and TFA (1 ml) was slowly added dropwise under ice bath. The reaction was continued for 1 h, and the reaction was monitored by TLC. The reaction solution was concentrated to obtain the crude product of compound 8.

[0247] Step 8: Dissolve the crude product of compound 8 from the previous step in 4 ml of DCM, slowly add DIPEA (200 μL, 1.21 mmol), and add acryloyl chloride (49 μL, 604 μmol) dropwise under ice bath. Continue the reaction for 10 min, and monitor the reaction by TLC. After the reaction is complete, add saturated NaHCO3, extract with EA (30 ml × 2), combine the organic phases, dry with anhydrous Na2SO4, filter, concentrate, and purify by preparative HPLC to obtain Example 5, LCMS (ESI) m / z 518.3.

[0248] Example 6

[0249]

[0250] The synthesis of (S)-1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-5Hpyrrolo[3,2-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 516.3 [M+H] + .

[0251] Example 7

[0252]

[0253] The synthesis of (S)-1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 516.3 [M+H] + .

[0254] Example 8

[0255]

[0256] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 532.3 [M+H] + .

[0257] Example 9

[0258]

[0259] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-7-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydropyrido[3,4-d]pyrimidin-8(5H)-one was described using the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0260] Example 10

[0261]

[0262] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(2-methyl-1,2,3,4-tetrahydroisoquinoline-8-yl)-1-((1-methylpyrrolidone-2-yl)methyl)-1,5,6,7-tetrahydro-2H-pyrrolo[3,4-d]pyrimidin-2-one is described in accordance with the general reaction procedure II-C; LCMS (ESI) m / z: 518.3 [M+H] + .

[0263] Example 11

[0264]

[0265] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(2-methyl-1,2,3,4-tetrahydroisoquinoline-8-yl)-1-((1-methylpyrrolidone-2-yl)methyl)-1,7-dihydro-2H-pyrrolo[2,3-d]pyrimidin-2-one is described in accordance with the general reaction procedure II-D; LCMS (ESI) m / z: 516.3 [M+H] + .

[0266] Example 12

[0267]

[0268] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(2-methyl-1,2,3,4-tetrahydroisoquinoline-8-yl)-1-((1-methylpyrrolidone-2-yl)methyl)-1,5-dihydro-2H-pyrrolo[3,2-d]pyrimidin-2-one is described in accordance with the general reaction procedure II-D; LCMS (ESI) m / z: 516.3 [M+H] + .

[0269] Example 13

[0270]

[0271] The synthesis of (S)-1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-yn-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 516.3 [M+H] + .

[0272] Example 14

[0273]

[0274] The synthesis of (S)-2-methyl-8-(2-((1-methylpyrrolidin-2-yl)methoxy)-4-(4-(ethylenesulfonyl)piperazin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)-1,2,3,4-tetrahydroisoquinoline is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 554.2 [M+H] + .

[0275] Example 15

[0276]

[0277] The synthesis of (S)-1-(7-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IB; LCMS (ESI) m / z: 556.3 [M+H] + .

[0278] Example 16

[0279]

[0280] The synthesis of (S)-1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpiperidin-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 530.3 [M+H] + .

[0281] Example 17

[0282]

[0283] The synthesis of 1-(4-(2-(dimethylamino)ethoxy)-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one was performed according to the general reaction procedure IA; LCMS (ESI) m / z: 492.3 [M+H] + .

[0284] Example 18

[0285]

[0286] The synthesis of 1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-(morpholinoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one was performed according to the general reaction procedure IA; LCMS (ESI) m / z: 520.3 [M+H] + .

[0287] Example 19

[0288]

[0289] The synthesis of 1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-(pyrrolidine-1-ylmethoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one was performed according to the general reaction procedure IA; LCMS (ESI) m / z: 504.3 [M+H] + .

[0290] Example 20

[0291]

[0292] The synthesis of 1-(4-(6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-3-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one was performed according to the general reaction procedure IA; LCMS (ESI) m / z: 518.3 [M+H] + .

[0293] Example 21

[0294]

[0295] The synthesis of (S)-1-(4-(5-methyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 532.3 [M+H] + .

[0296] Example 22

[0297]

[0298] The synthesis of (S)-1-(4-(5-cyclopropyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)propyl-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 558.3 ​​[M+H] + .

[0299] Example 23

[0300]

[0301] The synthesis of (S)-1-(4-(5,5-dimethyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0302] Example 24

[0303]

[0304] The synthesis of (S)-1-(4-(5-ethyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0305] Example 25

[0306]

[0307] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-5-isopropyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 574.3 [M+H] + .

[0308] Example 26

[0309]

[0310] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-5-isopropyl-6-(1-methyl-1,2,3,4-tetrahydroquinolin-5-yl)-2-(1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 574.3 [M+H] + .

[0311] Example 27

[0312]

[0313] The synthesis of (S)-5-methyl-4-(4-acryloylpiperazin-1-yl)-6-(1-methyl-1,2,3,4-tetrahydroquinolin-5-yl)-2-(1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0314] Example 28

[0315]

[0316] The synthesis of (S)-5-methyl-4-(4-acryloylpiperazin-1-yl)-6-(1-methyl-1,2,3,4-tetrahydroquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0317] Example 29

[0318]

[0319] The synthesis of (S)-1-(4-(5-isopropyl-6-(1-methyl-1,2,3,4-tetrahydroquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)propyl-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 560.3 [M+H] + .

[0320] Example 30

[0321]

[0322] The synthesis of (S)-1-(4-(6-(isochroman-5-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 505.2 [M+H] + .

[0323] Example 31

[0324]

[0325] The synthesis of (R)-4-(4-acryloylpiperazin-1-yl)-6-(2-methyl-1,2,3,4-tetrahydroisoquinoline-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 532.3 [M+H] + .

[0326] Example 32

[0327]

[0328] The synthesis of (S)-1-(4-(7-methyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 530.3 [M+H] + .

[0329] Example 33

[0330]

[0331] The synthesis of (S)-1-(4-(7-methyl-5-isopropyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)propyl-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 572.3 [M+H] + .

[0332] Example 34

[0333]

[0334] The synthesis of (S)-1-(4-(7-methyl-5-ethyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IB; LCMS (ESI) m / z: 558.3 ​​[M+H] + .

[0335] Example 35

[0336]

[0337] The synthesis of (S)-1-(4-(7-methyl-6-(8-chloronaphth-1-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 545.2 [M+H] + .

[0338] Example 36

[0339]

[0340] The synthesis of (S)-1-(4-(5-methyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-5Hpyrrolo[3,2-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 530.3 [M+H] + .

[0341] Example 37

[0342]

[0343] The synthesis of (S)-1-(4-(6-(isochroman-5-yl)-5,7-dimethyl-2-((1-methylpyrrolidin-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IB; LCMS (ESI) m / z: 531.3 [M+H] + .

[0344] Example 38

[0345]

[0346] The synthesis of (S)-1-(4-(7-methyl-5-amino-6-(isochroman-5-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IB; LCMS (ESI) m / z: 532.3 [M+H] + .

[0347] Example 39

[0348]

[0349] The synthesis of (S)-1-(4-(6-(2,3-dihydro-1H-inden-4-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 489.2 [M+H] + .

[0350] Example 40

[0351]

[0352] The synthesis of (R)-4-(4-acryloylpiperazin-1-yl)-5-methyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0353] Example 41

[0354]

[0355] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-5-methyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0356] Example 42

[0357]

[0358] The synthesis of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one was performed according to the general reaction procedure IA; LCMS (ESI) m / z: 546.3 [M+H] + .

[0359] Example 43

[0360]

[0361] The synthesis of (S)-1-(4-(5-methyl-6-(chroman-8-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-5Hpyrrolo[3,2-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 517.2 [M+H] + .

[0362] Example 44

[0363]

[0364] The synthesis of (S)-1-(4-(6-(chroman-5-yl)-5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-5Hpyrrolo[3,2-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 517.2 [M+H] + .

[0365] Example 45

[0366]

[0367] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-2-(1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 531.3 [M+H] + .

[0368] Example 46

[0369]

[0370] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-5-methyl-6-(8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-2-(1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one was described using the general reaction procedure IA; LCMS (ESI) m / z: 545.3 [M+H] + .

[0371] Example 47

[0372]

[0373] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(8,8-dimethyl-5,6,7,8-tetrahydronaphth-1-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one was described using the general reaction procedure IA; LCMS (ESI) m / z: 545.3 [M+H] + .

[0374] Example 48

[0375]

[0376] The synthesis of (S)-8-(4-(4-acryloylpiperazin-1-yl)-2-(1-methylpyrrolidine-2-yl)methoxy)-7-oxo-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)-1,2,3,4-tetrahydronaphthalene-1-carboxynitrile is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 542.2 [M+H] + .

[0377] Example 49

[0378]

[0379] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6-(5,6,7,8-tetrahydronaphth-1-yl)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one was described using the general reaction procedure IA; LCMS (ESI) m / z: 517.2 [M+H] + .

[0380] Example 50

[0381]

[0382] The synthesis of (S)-5-(4-(4-acryloylpiperazin-1-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)-3,4-dihydronaphthyl-2(1H)-one was performed according to the general reaction procedure IA; LCMS (ESI) m / z: 517.2 [M+H] + .

[0383] Example 51

[0384]

[0385] The synthesis of (S)-5-(4-(4-acryloylpiperazin-1-yl)-5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-5Hpyrrolo[3,2-d]pyrimidin-6-yl)-3,4-dihydronaphthyl-1(2H)-one was performed according to the general reaction procedure IB; LCMS (ESI) m / z: 529.2 [M+H] + .

[0386] Example 52

[0387]

[0388] The synthesis of (S)-8-(5-methyl-4-(4-acryloylpiperazin-1-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-5Hpyrrolo[3,2-d]pyrimidin-6-yl)-3,4-dihydronaphthyl-1(2H)-one was performed according to the general reaction procedure IB; LCMS (ESI) m / z: 529.2 [M+H] + .

[0389] Example 53

[0390]

[0391] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(8-fluoro-5,6,7,8-tetrahydronaphth-1-yl)-2-(1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one was described using the general reaction procedure IA; LCMS (ESI) m / z: 535.2 [M+H] + .

[0392] Example 54

[0393]

[0394] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-7-(8-methylnaphthyl-1-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydropyrido[3,4-d]pyrimidin-8(5H)-one was described using the general reaction procedure IA; LCMS (ESI) m / z: 541.28 [M+H] + .

[0395] Example 55

[0396]

[0397] The synthesis of (S)-4-(4-acryloylpiperazin-1-yl)-6-(8-methylnaphthyl-1-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 527.2 [M+H] + .

[0398] Example 56

[0399]

[0400] The synthesis of 4-(4-acryloylpiperazin-1-yl)-1-(2,4-dimethylpyridin-3-yl)-6-(2-fluoro-6-hydroxyphenyl)-5-methyl-1,5,6,7-tetrahydro-2H-pyrrolo[3,4-d]pyrimidin-2-one was performed according to the general reaction procedure II-C; LCMS (ESI) m / z: 505.2 [M+H] + .

[0401] Example 57

[0402]

[0403] The synthesis of 5-ethyl-4-(4-acryloylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-1,5,6,7-tetrahydro-2H-pyrrolo[3,4-d]pyrimidin-2-one was performed according to the general reaction procedure II-C; LCMS (ESI) m / z: 547.2 [M+H] + .

[0404] Example 58

[0405]

[0406] The synthesis of 5-methyl-4-(4-acryloylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-1,5,6,7-tetrahydro-2H-pyrrolo[3,4-d]pyrimidin-2-one was performed according to the general reaction procedure II-C; LCMS (ESI) m / z: 533.2 [M+H] + .

[0407] Example 59

[0408]

[0409] Synthesis of 5-methyl-4-(4-acryloylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-1,5-dihydro-2H-pyrrolo[2,3-d]pyrimidin-2-one: Refer to General Reaction Procedure II-D; LCMS (ESI) m / z: 531.2 [M+H] + .

[0410] Example 60

[0411]

[0412] The synthesis of (S)-1-(4-(5-cyclopropyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidone-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)-2-fluoroprop-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 576.3 [M+H] + .

[0413] Example 61

[0414]

[0415] The synthesis of (S)-1-(4-(5-isopropyl-6-(2-methyl-1,2,3,4-tetrahydroisoquinolin-8-yl)-2-((1-methylpyrrolidine-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)piperazin-1-yl)-2-fluoroprop-2-en-1-one is described in accordance with the general reaction procedure IA; LCMS (ESI) m / z: 578.3 [M+H] + .

[0416] Example 62

[0417]

[0418] The synthesis of 4-(4-acryloylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-1,7-dihydro-2H-pyrrolo[2,3-d]pyrimidin-2-one was performed according to the general reaction procedure II-D; LCMS (ESI) m / z: 517.2 [M+H] + .

[0419] Example 63

[0420]

[0421] The synthesis of (S)-1-(4-(7-methyl-8-(8-chloronaphth-1-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-7H-purin-6-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 546.2 [M+H] + .

[0422] Example 64

[0423]

[0424] The synthesis of (S)-1-(4-(7-methyl-8-(8-methylnaphthyl-1-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-7H-purin-6-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 526.2 [M+H] + .

[0425] Example 65

[0426]

[0427] The synthesis of (4-(7-methyl-6-(8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-7Hpyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 529.3 [M+H] + .

[0428] Example 66

[0429]

[0430] The synthesis of (4-acryloylpiperazin-1-yl)-6-(3-methyl-1,2,3,4-tetrahydroisoquinoline-8-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one was described using the general reaction procedure IA; LCMS (ESI) m / z: 532.3 [M+H] + .

[0431] Example 67

[0432]

[0433] The synthesis of (S)-1-(4-(7-methyl-8-(1H-indazol-4-yl)-2-((1-methylpyrrolidin-2-yl)methoxy)-7H-purin-6-yl)piperazin-1-yl)prop-2-en-1-one is described in the general reaction procedure IB; LCMS (ESI) m / z: 502.2 [M+H] + .

[0434] Example 68

[0435]

[0436] 2-(2S)-4-[6-(8-chloronaphthyl-1-yl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl]-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile, the synthesis of which is described in the synthetic route of Example 1 of the original patent, m / z 603.2.

[0437] Example 69

[0438]

[0439] 2-(2S)-4-[5-methyl-6-(8-chloronaphthyl-1-yl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl]-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile, the synthesis of which is described in the synthetic route of Example 1 of the original patent, m / z 617.2.

[0440] Example 70

[0441]

[0442] 2-(2S)-4-[5-(5R)methyl-6-(8-chloronaphthyl-1-yl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl]-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile, the synthesis of which is described in the synthetic route of Example 1 of the original patent, m / z 617.2.

[0443] Example 71

[0444]

[0445] 2-(2S)-4-[5-(5S)-methyl-6-(8-chloronaphthyl-1-yl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl]-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile, the synthesis of which is described in the synthetic route of Example 1 of the original patent, m / z 617.2.

[0446] Example 72

[0447]

[0448] 2-(2S)-4-[5-(5S)-methyl-6-(8-chloronaphthyl-1-yl)-2-(((R)-1-methylpyrrolidine-2-yl)methoxy)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl]-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile, the synthesis of which is described in the synthetic route of Example 1 of the original patent, m / z 617.2.

[0449] Example 73

[0450]

[0451] 2-(2S)-4-[5-(5R)-methyl-6-(8-chloronaphthyl-1-yl)-2-(((R)-1-methylpyrrolidine-2-yl)methoxy)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl]-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile, the synthesis of which is described in the synthetic route of Example 1 of the original patent, m / z 617.2.

[0452] Example 74

[0453]

[0454] 2-(2R)-4-[5-(5S)-methyl-6-(8-chloronaphthyl-1-yl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl]-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile, the synthesis of which is described in the synthetic route of Example 1 of the original patent, m / z 617.2.

[0455] Experimental Example A

[0456] Determination of KRAS G12C covalent bonding

[0457] The covalent binding of the compound in the examples to the KRAS G12C mutant protein in cells was determined using gel mobility shift assay.

[0458] Three human non-small cell lung cancer cell lines were used: NCI-H358 cells (KRAS G12C mutation), A549 cells (KRAS G12S mutation), and HCC827 cells (KRAS wild type).

[0459] Experimental reagents and instruments: RPMI 1640 medium, DMEM cell culture medium, fetal bovine serum, 0.25% trypsin-0.53mM EDTA digestion solution, DMSO, penicillin-streptomycin, KRAS antibody (Sigma), secondary antibody: Anti-rabbit IgG-HRP, Cell Titer-Gio assay kit. Promega microplate reader, cell culture flasks, cell culture microplates (96 or 384 wells), CO2 incubator, FluorChemR analyzer (ProteinSimple).

[0460] The electrophoretic migration changes of the covalent complex formed by the compound and the KRAS G12C mutant protein were determined using gel mobility shift assay. Based on the increased molecular weight of the covalent complex formed by the compound and the KRAS G12C mutant protein, a corresponding band lag was observed during electrophoresis compared to the unbound KRAS G12C mutant protein. The electrophoretic bands were quantitatively scanned using a FluorChemR detector, and the electrophoretic migration ratio of the compound-KRAS G12C mutant protein complex to the unbound G12C mutant protein was calculated, providing a rapid and direct determination of the covalent binding characteristics of the compound to the KRAS G12C mutant protein. The greater the binding of the compound to the KRAS G12C mutant protein, the larger the calculated ratio.

[0461] Cell culture: After reviving tumor cells frozen in liquid nitrogen, the cells were cultured in cell culture medium containing 10% fetal bovine serum and 10% penicillin-streptomycin. When the cells grew to the exponential growth phase, they were digested, centrifuged, and resuspended in culture medium. Cells were seeded at a rate of 5,000-10,000 cells per well and cultured in a carbon dioxide incubator at a constant temperature of 37°C, 5% CO2, and saturated humidity.

[0462] Cells were treated with different concentrations of compounds for different durations. Cells were washed with phosphate-buffered saline (PBS), then lysed in RIPA buffer (50 mm Tris, pH 7.5, 150 mm NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, containing protease and phosphatase inhibitors). Total cellular protein was extracted, and protein concentration was determined using a BCA protein assay kit. Equal volumes of protein were subjected to SDS-PAGE electrophoresis. After electrophoresis, the gel was transferred to a nitrocellulose membrane. The membrane was blocked in TBS-0.1% Tween-20 with 5% skim milk, then primary antibody was added, and the membrane was gently shaken overnight at 4°C. After overnight incubation, the membrane was washed and incubated with fluorescently bound secondary antibody for 1 hour at room temperature. The electrophoretic bands were then scanned using a FluorChemR detector (ProteinSimple).

[0463] In gel migration analysis, the KRAS G12C inhibitor of this invention selectively forms a covalent complex with the KRAS G12C mutant protein in human non-small cell lung cancer NCI-H358 cells containing the KRAS G12C mutation. The resulting compound-G12C mutant protein covalent complex exhibits gel migration arrest. However, the compound cannot form a covalent complex with KRAS G12S mutant non-small cell lung cancer A549 cells, nor with wild-type KRAS-containing non-small cell lung cancer HCC827 cells (Table 1). The experimental results are expressed as follows: "A" ≤ 25% ratio; "B" > 25% - ≤ 50%; "C" ≥ 50%.

[0464] Table 1. Determination of the covalent complex of the compounds in the examples with the KRAS G12C mutant protein.

[0465]

[0466]

[0467] Experimental Example B

[0468] Assay for the inhibitory activity of compounds on cell proliferation

[0469] In this experimental example, the antiproliferative activity of the exemplary compound against KRAS G12C mutant tumor cells was determined using the Luminometer luminescence assay.

[0470] Three human non-small cell lung cancer cell lines were used: NCI-H358 cells (KRAS G12C mutation), A549 cells (KRAS G12S mutation), and HCC827 cells (KRAS wild type).

[0471] Experimental reagents and instruments: RPMI 1640 medium, DMEM cell culture medium, fetal bovine serum, 0.25% trypsin-0.53mM EDTA digestion solution, DMSO, penicillin-streptomycin, Cell Titer-Gio assay kit, Promega microplate reader, cell culture flasks, cell culture microplates (96 or 384 wells), CO2 incubator.

[0472] Cell culture: Resuscitate tumor cells frozen in liquid nitrogen, culture the cells in cell culture medium containing 10% fetal bovine serum and 10% penicillin-streptomycin, and when the cells grow to the exponential growth phase, digest and centrifuge to collect the cells and resuspend them in the culture medium; seed the cells at a number of 5,000-10,000 cells per well and incubate overnight in a constant temperature incubator at 37°C, 5% CO2, and saturated humidity.

[0473] After 72 hours of compound treatment, the 96-well plates were removed from the 37°C incubator and placed at room temperature for 30 minutes for CTG detection. The plates were not shaken during the experiment. 100 μl of CTG reagent was added, mixed for 2 minutes, and then incubated at room temperature for 10 minutes. The luminescence values ​​were detected and recorded using a GloMax 96-well microplate chemiluminescence analyzer (CellTiter-Glo Luminescent Cell Viability Assay, Promega) to observe cell viability.

[0474] Each test compound of the present invention was diluted into 10 concentration gradients and added to the corresponding wells of a cell plate. The cell plates were then returned to a CO2 incubator for further incubation for 72 hours. After incubation, Promega CellTiter-Glo reagent was added to each well of the cell plate, and the plates were incubated at room temperature for 10 minutes. The luminescence signal was detected using a Promega microplate reader, and the IC50 value was calculated. The results of the antiproliferative activity of the compounds of the present invention are expressed as A, B, and ND: "A" ≥ 0.001 - ≤ 1 μM; "B" > 1 μM; ND = Not determined.

[0475] The compounds of this invention exhibited high antiproliferative activity against KRAS G12C-mutant human non-small cell lung cancer NCL-H358 cells, but weaker antiproliferative activity against KRAS G12S-mutant human non-small cell lung cancer A549 cells and KRAS wild-type HCC827 cells (Table 2). The antiproliferative assay results were consistent with those obtained using the gel migration assay described above, demonstrating the high selectivity of the compounds of this invention against KRAS G12C-mutant tumor cells.

[0476] Table 2. Antiproliferative effects of compounds in the examples on KRAS-G12C mutant tumor cells.

[0477]

[0478]

[0479] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A compound represented by Formula III-1 or a pharmaceutically acceptable salt thereof: wherein: ； ； R2 is hydrogen; R 10 is naphthyl or tetrahydroisoquinolinyl; R 15 is optionally substituted by 1 or more, same or different R 14 is optionally substituted by 1 or more, same or different R 15 is tetrahydropyrrolylmethyl; R 15 is hydrogen or C1-C6alkyl; R 16 is hydrogen; R 18 is hydrogen; J is O.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, the C1-C6 alkyl is selected from the group consisting of methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl and hexyl.

3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 14 is tetrahydropyrrolylmethyl optionally substituted by one or more methyl groups.

4. The compound or pharmaceutically acceptable salt thereof of claim 1, wherein, R 10 is naphthyl or tetrahydroisoquinolinyl optionally substituted with 1 or more methyl groups.

5. A compound represented by the following formula or a pharmaceutically acceptable salt thereof: 。 6. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 and an excipient.

7. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, or the pharmaceutical composition according to claim 6, in the manufacture of a medicament for treating a KRAS G12C mutation-related tumor.

8. The use according to claim 7, wherein, The KRAS G12C mutation-related tumor includes a tumor occurring at the following sites: lung, colorectal, pancreas, brain, head and neck, liver, stomach, esophagus, breast, cervix, ovary, endometrium, prostate, thyroid and soft tissue.

9. The use according to claim 7, wherein, The KRAS G12C mutation-related tumor includes a tumor occurring at the following sites: larynx, oral cavity.

10. Use according to claim 7, wherein, The tumor includes: lung cancer, pancreatic cancer, gastric cancer, esophageal cancer, small intestine cancer, kidney cancer, bladder cancer, urethral cancer, prostate cancer, testicular cancer, hepatocellular carcinoma, cholangiocarcinoma, brain astrocytoma, retinoblastoma, endometrial cancer, cervical cancer, ovarian cancer, sarcoma, acute lymphoblastic leukemia, chronic lymphocytic leukemia, multiple myeloma, myelodysplastic syndrome, Hodgkin's disease, non-Hodgkin's lymphoma, malignant melanoma, hemangioma.

11. Use according to claim 7, wherein, The tumor includes: non-small cell lung cancer, small cell lung cancer, lung squamous cell carcinoma, pancreatic ductal adenocarcinoma, pancreatic islet tumor, esophageal squamous cell carcinoma, esophageal adenocarcinoma, esophageal leiomyosarcoma, small intestine adenocarcinoma, kidney adenocarcinoma, Wilms' tumor, bladder and urethral squamous cell carcinoma, bladder and urethral adenocarcinoma, prostate sarcoma, seminoma, brain glioblastoma, osteosarcoma, fibrosarcoma, chondrosarcoma.

12. A composition comprising the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, and another antitumor drug.

13. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, or the pharmaceutical composition according to claim 6, or the composition according to claim 12, for non-therapeutic inhibition of KRAS G12C mutant protein.

14. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, or the pharmaceutical composition according to claim 6, or the composition according to claim 12, in the manufacture of a medicament for inhibiting KRAS G12C mutant protein.

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