Spirocyclic compounds and their applications in medicine

Abstract

The present invention provides a class of spirocyclic compounds and their application in medicine, belonging to the field of medicine, and specifically relates to KRAS mutant inhibitors. The spirocyclic compounds provided by the present invention have an inhibitory effect on KRAS G12D and KRAS G12V Mutated tumor cells have a good inhibitory effect.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to KRAS mutant inhibitors. Background Art

[0002] The human RAS gene family includes three types of RAS genes, KRAS, NRAS, and HRAS, encoding four different RAS proteins (KRAS-4A, KRAS-4B, NRAS, and HRAS). RAS proteins belong to the GTPase protein family. They are inactive when bound to GDP, but active when bound to GTP, which can lead to the activation of downstream signaling pathways such as RAF-MAPK and PI3K-Akt, resulting in cell anti-apoptosis and proliferation (Cell, 2017; 170(1): 17-33; Cell, 2020; 183(4): 850-859; Nat Rev Drug Discov, 2020; 19(8): 533-552.). Activating mutations of RAS genes are the most common oncogenic driver genes in human cancers, among which KRAS is the most common oncogenic activating mutation.

[0003] Oncogenic driver mutations can occur at multiple sites in the KRAS gene, with the most common mutations occurring at the G12 site, including G12C, G12D, G12V, etc. These mutations can reduce the GTPase activity of the KRAS protein, causing the KRAS protein to be active for a long time, leading to malignant transformation of cells and cancer (Cell, 2017; 170(1): 17-33; Nat Rev Drug Discov, 2020; 19(8): 533-552). The types of KRAS mutations that frequently occur in different types of cancer vary.

[0004] The affinity of KRAS protein for GDP / GTP is at the picomolar level, while the affinity of ATP-dependent protein kinase for ATP is only at the micromolar level. This difference makes it extremely difficult for compound molecules to effectively compete with GDP / GTP, which seriously hinders the development of KRAS inhibitors (Cell, 2017; 170 (1): 17-33; Cell, 2020; 183 (4): 850-859; Nat Rev Drug Discov, 2020; 19 (8): 533-552). In recent years, researchers have discovered an "allosteric" binding cavity on the KRAS protein that can effectively compete with small molecules for non-GDP / GTP binding. This discovery has greatly promoted the development of targeted drugs for KRAS mutation-driven tumors. Currently, MRTX-849 (Adagrasib) and AMG510 (Sotorasib) targeting KRAS G12C have shown excellent efficacy in clinical studies (N Engl J Med. 2020; 383 (13): 1207-1217; Cancer Discov. 2020; 10 (1): 54-71), and AMG510 and MRTX-849 have been successfully approved by the FDA for marketing. However, compared with the research and development results of KRASG12C drugs, the research and development of drugs targeting KRAS G12D, G13D, G12V, G12R and other mutations is still in its infancy, and the number of molecules entering the clinical stage is relatively small. The research and development of pan-KRAS mutation inhibitors is urgent to meet the actual clinical needs. Summary of the invention

[0005] In one aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, isotopic variant, tautomer or stereoisomer thereof: (I) in, Z is selected from O, S, SO2, CH2CH2, CH2 or NH; Z1, Z2, Z3 are each independently selected from O, S or CH2; X1 is selected from CH or N; Y is selected from CH, CF, CMe or N; R is selected from -N(R a )(R a '),OR a , C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-12 membered heterocyclic group, or ; R1 is selected from H, C 1-6 Alkyl or 4-12 membered heterocyclic group; R and R1 are independently replaced by 1-3 R x replace; R x Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkenyl, halogen, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Alkoxy, -(CH2) p CN, NH2, OH, -O(CH2) p OR a 、-O(CH2) p N(R a )(R a '),-N(R a )-(CH2) p OR a 、-O(CH2) p -R a 、-C(O)R a 、-C(O)OR a 、-(CH2) p -OR a 、-SR a 、-S(O)2-R a 、-(CH2) p -C(O)OR a 、-(CH2) p -OC(O)N(R a )(R a '),-N(R a )-S(O)2R a ', -N(R a )-S(O)R a '、-OC(O)N(R a )(R a '); R3 is selected from -NR m R n or a 4-12 membered heterocyclic group; R m Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, CN, -LC(O)R a 、-LC(O)OR a 、-(CH2)p -OR a 、-S(O)2-R a , -L-5-12 membered heteroaryl, -L-phenyl, -L-5-6 membered heterocyclic group; the R m Optionally 1-6 halogens, NH2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, CN, -L-OR a 、-L-SR a 、-LN(R a )(R a '), -NH-(CH2) p -C(O)OR a , C 3-6 further substituted by cycloalkyl, 4-6 membered heterocyclyl or 5-12 membered heteroaryl; R n Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl or C 3-6 Cycloalkyl; When R m When selected from -L-5-12 membered heteroaryl, the 5-12 membered heteroaryl may be further substituted by 1-3 R p replace; R p Selected from halogen, NH2, OH, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or CN; L is selected from a bond, C 1-6 Alkyl or C 1-6 L may be optionally substituted by halogen, OH or C 1-6 Alkoxy substitution; R a Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl, 4-6 membered heterocyclic group, C 3-6 Cycloalkyl or 5-6 membered heteroaryl; R a 'Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl 4-6 membered heterocyclic group or C 3-6 Cycloalkyl; p is independently selected from 0, 1, 2, 3, 4 or 5.

[0006] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, and optionally a pharmaceutically acceptable excipient.

[0007] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient, which may also contain another therapeutic agent.

[0008] In another aspect, the present invention provides use of a compound of the present invention in the preparation of a medicament for treating and / or preventing a disease mediated by a KRAS mutant.

[0009] In another aspect, the present invention provides a method for treating and / or preventing a KRAS mutant-mediated disease in a subject, comprising administering to the subject a compound or composition of the present invention.

[0010] In another aspect, the present invention provides a compound of the present invention or a composition of the present invention for use in the treatment and / or prevention of a KRAS mutant-mediated disease.

[0011] In a specific embodiment, the diseases treated by the present invention include cancers selected from the group consisting of acoustic neuroma, adenocarcinoma, adrenal cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendothelioma, hemangioma), appendix cancer, benign monoclonal gamma disease, bile duct cancer, bladder cancer, brain cancer (e.g., meningioma, glioma, e.g., astrocytoma, oligodendroglioma, medulloblastoma), bronchial cancer, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial cancer, ependymoma, endothelial sarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma), Ewing's sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma), hypereosinophilia, gallbladder cancer, stomach cancer (e.g., gastric adenocarcinoma), gastrointestinal stromal tumors (GIST), head and neck cancer (e.g., head and neck squamous cell carcinoma), oral cancer (e.g., oral squamous cell carcinoma), laryngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal carcinoma, oropharyngeal cancer)), hematopoietic system cancer (e.g., leukemia, such as acute lymphoblastic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), Acute myeloid leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myeloid leukemia (CML) (e.g., B-cell CML, T-cell CML), chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), marginal zone B-cell lymphoma (e.g., mucosa-associated lymphoid tissue (MALT) lymphoma, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoma Plasma cell lymphoma, hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell non-Hodgkin lymphoma, such as precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (such as cutaneous T-cell lymphoma (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); mixtures of one or more of the above leukemias / lymphomas;Multiple myeloma (MM), heavy chain disease (eg, alpha chain disease, gamma chain disease, mu chain disease), hemangioblastoma, inflammatory myofibroblastic tumor, immune cell amyloidosis, kidney cancer (eg, Wilms tumor, renal cell carcinoma), liver cancer (eg, hepatocellular carcinoma, malignant hepatocellular carcinoma), lung cancer (eg, bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma, leiomyosarcoma (LMS), mastocytosis (eg, systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disorders (MPD) (eg, true Polycythemia vera (PV), essential thrombocythemia (ET), idiopathic myeloid metaplasia (AMM), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES), neuroblastoma, neurofibroma (such as type 1 or type 2 neurofibromatosis, neurothecoma), neuroendocrine cancer (such as gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer (such as cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma, penile cancer. ;

[0012] definition Chemical Definition Definitions of specific functional groups and chemical terms are described in more detail below.

[0013] When a numerical range is listed, it is intended to include every value and sub-range within the stated range. For example, "C 1-6 "Alkyl" includes C1, C2, C3, C4, C5, C6, C 1-6 , C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-5 , C 2-4 , C 2-3 , C 3-6 , C 3-5 , C 3-4 , C 4-6 , C 4-5 and C 5-6 alkyl.

[0014] “C 1-6 "Alkyl" refers to a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms. In some embodiments, C 1-4 Alkyl and C 1-2 Alkyl groups are preferred. 1-6Examples of alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). The term “C 1-6 "Alkyl" also includes heteroalkyl groups in which one or more (e.g., 1, 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkyl group may be optionally substituted by one or more substituents, for example, by 1 to 5 substituents, 1 to 3 substituents or 1 substituent. Conventional alkyl abbreviations include: Me(-CH3), Et(-CH2CH3), iPr(-CH(CH3)2), nPr(-CH2CH2CH3), n-Bu(-CH2CH2CH2CH3) or i-Bu(-CH2CH(CH3)2).

[0015] “C 2-6 "Alkenyl" refers to a straight or branched hydrocarbon group having 2 to 6 carbon atoms and at least one carbon-carbon double bond. In some embodiments, C 2-4 Alkenyl is preferred. 2-6 Examples of alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. The term “C 2-6 "Alkenyl" also includes heteroalkenyl groups in which one or more (e.g., 1, 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkenyl group may be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0016] “C 2-6 "Alkynyl" refers to a straight or branched chain hydrocarbon group having 2 to 6 carbon atoms, at least one carbon-carbon triple bond, and optionally one or more carbon-carbon double bonds. In some embodiments, C 2-4 Alkynyl is preferred. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), pentynyl (C5), hexynyl (C6), and the like. The term “C 2-6"Alkynyl" also includes heteroalkynyl groups, in which one or more (e.g., 1, 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). Alkynyl groups may be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0017] “C 1-6 "Alkylene" refers to the removal of C 1-6 The other hydrogen of C alkyl forms a divalent group, and may be substituted or unsubstituted. 1-4 Alkylene, C 2-4 Alkylene and C 1-3 Alkylene is preferred. Unsubstituted alkylene includes, but is not limited to, methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-), pentylene (-CH2CH2CH2CH2CH2-), hexylene (-CH2CH2CH2CH2CH2CH2-), and the like. Exemplary substituted alkylenes, for example, alkylenes substituted with one or more alkyl (methyl) groups, include, but are not limited to, substituted methylene (-CH(CH3)-, -C(CH3)2-), substituted ethylene (-CH(CH3)CH2-, -CH2CH(CH3)-, -C(CH3)2CH2-, -CH2C(CH3) 2- ), substituted propylene (-CH(CH3)CH2CH2-, -CH2CH(CH3)CH2-, -CH2CH2CH(CH3)-, -C(CH3)2CH2CH2-, -CH2C(CH3)2CH2-, -CH2CH2C(CH3)2-), and the like.

[0018] “C 2-6 "Alkenylene" means removing the C 2-6 The divalent group is formed by the other hydrogen of the alkenyl group and may be substituted or unsubstituted. 2-4Alkenylene is particularly preferred. Exemplary unsubstituted alkenylene includes, but is not limited to, vinylene (-CH=CH-) and propenylene (e.g., -CH=CHCH2-, -CH2-CH=CH-). Exemplary substituted alkenylene, for example, alkenylene substituted with one or more alkyl (methyl) groups, includes, but is not limited to, substituted ethylidene (-C(CH3)=CH-, -CH=C(CH3)-), substituted propenylene (-C(CH3)=CHCH2-, -CH=C(CH3)CH2-, -CH=CHCH(CH3)-, -CH=CHC(CH3)2-, -CH(CH3)-CH=CH-, -C(CH3)2-CH=CH-, -CH2-C(CH3)=CH-, -CH2-CH=C(CH3)-), etc.

[0019] “C 2-6 "Alkynylidene" refers to the removal of C 2-6 The other hydrogen of the alkynyl group forms a divalent group, and may be substituted or unsubstituted. 2-4 Alkyne is particularly preferred. Exemplary alkynyl includes, but is not limited to, ethynyl (-C≡C-), substituted or unsubstituted propynyl (-C≡CCH2-), and the like.

[0020] "Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0021] Therefore, “C 1-6 "Haloalkyl" refers to the above-mentioned "C 1-6 "alkyl", which is substituted by one or more halogen groups. In some embodiments, C 1-4 Haloalkyl is particularly preferred, more preferably C 1-2 Haloalkyl. Exemplary haloalkyls include, but are not limited to, -CF3, -CH2F, -CHF2, -CHFCH2F, -CH2CHF2, -CF2CF3, -CCl3, -CH2Cl, -CHCl2, 2,2,2-trifluoro-1,1-dimethyl-ethyl, and the like. The haloalkyl group may be substituted at any available point of attachment, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0022] “C 1-6 "Alkoxy" refers to an -OR group, wherein R is a C 1-6 Alkyl. C 1-4 Alkoxy groups are preferred.

[0023] “C 1-6 "Haloalkoxy" refers to "C 1-6Alkoxy", which is substituted by one or more halogen groups. In some embodiments, C 1-4 Halogenated alkoxyalkyl is particularly preferred, more preferably C 1-2 Halogenated alkoxyalkyl.

[0024] “C 3-10 "Cycloalkyl" refers to a non-aromatic cyclic hydrocarbon group having 3 to 10 ring carbon atoms and zero heteroatoms. In some embodiments, C 4-10 Cycloalkyl, C 5-10 Cycloalkyl, C 4-7 Cycloalkyl, C 3-7 Cycloalkyl, C 3-6 Cycloalkyl, C 3-5 Cycloalkyl and C 3-4 Cycloalkyl is particularly preferred, more preferably C 5-6 Cycloalkyl. Cycloalkyl also includes a ring system in which the above-mentioned cycloalkyl ring is fused with one or more aryl or heteroaryl groups, wherein the connection point is on the cycloalkyl ring, and in such a case, the number of carbons continues to represent the number of carbons in the cycloalkyl system. Exemplary cycloalkyls include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), etc. The cycloalkyl group may be optionally substituted by one or more substituents, for example, substituted by 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0025] "3-12 membered heterocyclyl" refers to a group of a 3- to 12-membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon. In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as the valence permits. In some embodiments, a 3-10 membered heterocyclyl is preferably a 3-10 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; in some embodiments, a 4-10 membered heterocyclyl is preferably a 4-10 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms; in some embodiments, a 5-10 membered heterocyclyl is preferably a 5-10 membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 5-8 membered heterocyclyl is preferably a 5-8 membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 3-7 membered heterocyclyl is preferably a A 3- to 7-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms; preferably a 3- to 6-membered heterocyclyl, which is a 3- to 6-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; preferably a 4- to 7-membered heterocyclyl, which is a 4- to 7-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; preferably a 4- to 6-membered heterocyclyl, which is a 4- to 6-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; more preferably a 5- to 6-membered heterocyclyl, which is a 5- to 6-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; more preferably a 3- to 5-membered heterocyclyl, which is a 3- to 5-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms. Heterocyclyl also includes a ring system in which the above-mentioned heterocyclyl ring is fused to one or more cycloalkyl groups, wherein the point of attachment is on the cycloalkyl ring, or a ring system in which the above-mentioned heterocyclyl ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring; and in such a case, the number of ring members continues to represent the number of ring members in the heterocyclyl ring system. Exemplary 3-membered heterocyclyls containing one heteroatom include, but are not limited to, aziridine, oxirane, and thiorenyl. Exemplary 4-membered heterocyclyls containing one heteroatom include, but are not limited to, azetidine, oxirane, and thiorenyl. Exemplary 5-membered heterocyclyls containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl.Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to, hexahydrotriazine (triazinanyl). Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azepanyl, oxepane, and thianyl. Exemplary 5-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 5,6-bicyclic heterocyclic groups) include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinone, and the like. Exemplary 6-membered heterocyclyl groups fused to a C6 aryl ring (also referred to herein as 6,6-bicyclic heterocyclyl groups) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydrobenzopyranyl, tetrahydropyranopyridinyl, etc. The heterocyclyl group may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0026] “C 6-10 "Aryl" refers to a monocyclic or polycyclic (e.g., bicyclic) 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having 6-10 ring carbon atoms and zero heteroatoms. In some embodiments, aryl has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, aryl has ten ring carbon atoms ("C 10 Aryl also includes ring systems in which the above aryl ring is fused to one or more cycloalkyl or heterocyclyl groups, and the point of attachment is on the aryl ring, in which case the number of carbon atoms continues to refer to the number of carbon atoms in the aryl ring system. Aryl groups can be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0027] "5-14 membered heteroaryl" refers to a group of a 5-14 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic arrangement) having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as the valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings. Heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl or heterocyclic groups, and the point of attachment is on the heteroaryl ring, in which case the number of carbon atoms continues to represent the number of carbon atoms in the heteroaryl ring system. In some embodiments, a 5-10 membered heteroaryl is preferred, which is a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms. In some embodiments, 5-10 membered heteroaryls are preferred, which are 6-10 membered monocyclic or bicyclic 4n+2 aromatic ring systems with ring carbon atoms and 1-4 ring heteroatoms. In some embodiments, 5-9 membered heteroaryls are preferred, which are 5-9 membered monocyclic or bicyclic 4n+2 aromatic ring systems with ring carbon atoms and 1-4 ring heteroatoms. In other embodiments, 5-6 membered heteroaryls are particularly preferred, which are 5-6 membered monocyclic or bicyclic 4n+2 aromatic ring systems with ring carbon atoms and 1-4 ring heteroatoms. Exemplary 5-membered heteroaryls containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5-membered heteroaryls containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl) and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azacycloheptatrienyl, oxacycloheptatrienyl and thiacycloheptatrienyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indazinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.A heteroaryl group can be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0028] "Cycloalkylene", "heterocyclylene", "arylene" or "heteroarylene" is a divalent group formed by removing another hydrogen from the above-defined "cycloalkyl", "heterocyclyl", "aryl" or "heteroaryl", and may be substituted or unsubstituted. For example, "C 5-7 "Cycloalkylene" refers to the removal of C 5-7 The "5- to 8-membered heterocyclylene" refers to a divalent group formed by removing another hydrogen atom of a 5- to 8-membered heterocyclyl. 6-10 "Arylene" refers to the removal of C 6-10 The "5-6 membered heteroarylene group" refers to a divalent group formed by removing another hydrogen atom of a 5-6 membered heteroaryl group.

[0029] Alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups and the like as defined herein are optionally substituted groups.

[0030] Exemplary substituents on carbon atoms include, but are not limited to, halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR aa 、-ON(R bb )2、-N(R bb )2、-N(R bb )3 + X - 、-N(OR cc )R bb , -SH, -SR aa 、-SSR cc 、-C(=O)R aa 、-CO2H、-CHO、-C(OR cc 2. -CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(R bb )2、-OC(=O)N(R bb )2.-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2、-C(=O)NR bb SO2R aa 、-NR bb SO2R aa 、-SO2N(R bb )2、-SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3、-OSi(R aa )3、-C(=S)N(R bb )2、-C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa 、-OC(=O)SR aa 、-SC(=O)OR aa 、-SC(=O)R aa 、-P(=O)2R aa 、-OP(=O)2R aa 、-P(=O)(R aa )2、-OP(=O)(R aa )2、-OP(=O)(OR cc )2、-P(=O)2N(R bb )2、-OP(=O)2N(R bb )2、-P(=O)(NR bb )2、-OP(=O)(NR bb )2、-NR bb P(=O)(OR cc )2、-NR bb P(=O)(NR bb )2、-P(R cc )2、-P(R cc )3、-OP(R cc )2、-OP(Rcc )3. -B(R aa )2、-B(OR cc )2. -BR aa (OR cc ), alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substitution; Or the two geminal hydrogen atoms on the carbon atom are replaced by groups =O, =S, =NN(R bb )2、=NNR bb C(=O)R aa 、=NNR bb C(=O)OR aa 、=NNR bb S(=O)2R aa 、=NR bb =NOR cc replace; R aa Each of R is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R aa The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substitution; R bb Each of the following is independently selected from: hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)R aa 、-C(=O)N(R cc 2. -CO2R aa 、-SO2R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc 2. -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2. -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)2R aa 、-P(=O)(R aa )2、-P(=O)2N(Rcc )2、-P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R bb The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution; R cc Each of R is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R cc The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution; R dd Each of is independently selected from: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee 、-ON(R ff )2、-N(R ff )2, 、-N(R ff )3 + X - 、-N(OR ee )R ff , -SH, -SR ee 、-SSR ee 、-C(=O)R ee 、-CO2H、-CO2R ee 、-OC(=O)R ee 、-OCO2R ee 、-C(=O)N(R ff )2、-OC(=O)N(R ff )2.-NR ff C(=O)R ee 、-NR ff CO2R ee 、-NR ff C(=O)N(R ff )2、-C(=NR ff )OR ee 、-OC(=NR ff )R ee 、-OC(=NR ff )OR ee 、-C(=NR ff )N(R ff )2. -OC(=NR ff )N(Rff )2.-NR ff C(=NR ff )N(R ff )2.-NR ff S02R ee 、-SO2N(R ff 2. -SO2R ee 、-SO2OR ee 、-OSO2R ee 、-S(=O)R ee 、-Si(R ee )3.-OSi(R ee )3. -C(=S)N(R ff )2. -C(=O)SR ee 、-C(=S)SR ee 、-SC(=S)SR ee 、-P(=O)2R ee 、-P(=O)(R ee )2、-OP(=O)(R ee )2、-OP(=O)(OR ee )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg group substitution, or two geminal R dd Substituents may combine to form =O or =S; R ee Each of R is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg group substitution; R ff Each of R is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R ff The groups are combined to form a heterocyclic or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 R gg group substitution; R gg Each of is independently: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC 1-6 Alkyl, -ON(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)2, -N(C 1-6Alkyl)3 + X - 、-NH(C 1-6 Alkyl)2 + X - 、-NH2(C 1-6 alkyl) + X - 、-NH3 + X - 、-N(OC 1-6 Alkyl)(C 1-6 Alkyl), -N(OH)(C 1-6 Alkyl), -NH(OH), -SH, -SC 1-6 Alkyl, -SS(C 1-6 Alkyl), -C(=O)(C 1-6 alkyl), -CO2H, -CO2(C 1-6 Alkyl), -OC(=O)(C 1-6 Alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 Alkyl)2, -OC(=O)NH(C 1-6 Alkyl), -NHC(=O)(C 1-6 Alkyl), -N(C 1-6 Alkyl)C(=O)(C 1-6 Alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 Alkyl)2, -NHC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -C(=NH)O(C 1-6 Alkyl), -OC(=NH)(C 1-6 Alkyl), -OC(=NH)OC 1-6 Alkyl, -C(=NH)N(C 1-6 Alkyl)2, -C(=NH)NH(C 1-6 alkyl), -C(=NH)NH2, -OC(=NH)N(C 1-6 Alkyl)2, -OC(NH)NH(C 1-6 alkyl), -OC(NH)NH2, -NHC(NH)N(C 1-6 Alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1-6 Alkyl), -SO2N(C 1-6 Alkyl)2, -SO2NH(C 1-6 Alkyl), -SO2NH2, -SO2C 1-6 Alkyl, -SO2OC 1-6Alkyl, -OSO2C 1-6 Alkyl, -SOC 1-6 Alkyl, -Si(C 1-6 Alkyl)3, -OSi(C 1-6 Alkyl)3, -C(=S)N(C 1-6 Alkyl)2, C(=S)NH(C 1-6 alkyl), C(=S)NH2, -C(=O)S(C 1-6 Alkyl), -C(=S)SC 1-6 Alkyl, -SC(=S)SC 1-6 Alkyl, -P(=O)2(C 1-6 Alkyl), -P(=O)(C 1-6 Alkyl)2, -OP(=O)(C 1-6 Alkyl)2, -OP(=O)(OC 1-6 Alkyl)2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, C6-C 10 Aryl, C3-C7 heterocyclic, C5-C 10 heteroaryl; or two geminal R gg Substituents may combine to form =O or =S; where X - For the counter ion.

[0031] Exemplary substituents on nitrogen atoms include, but are not limited to, hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)R aa 、-C(=O)N(R cc 2. -CO2R aa 、-SO2R aa 、-C(=NR bb )R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc 2. -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2. -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)2R aa 、-P(=O)(R aa )2、-P(=O)2N(Rcc )2、-P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R attached to the nitrogen atom cc The groups are combined to form a heterocyclic or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 R dd substituted with a group, and wherein R aa , R bb , R cc and R dd As mentioned above.

[0032] Other definitions The term "pharmaceutically acceptable salt" as used herein refers to those carboxylates, amino acid addition salts of the compounds of the present invention which are suitable for use in contact with patient tissues within the scope of sound medical judgment, do not produce undue toxicity, irritation, allergic response, etc., are commensurate with a reasonable benefit / risk ratio, and are effective for their intended use, including (where possible) zwitterionic forms of the compounds of the present invention.

[0033] "Subjects" for administration include, but are not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or older adults)) and / or non-human animals, e.g., mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein.

[0034] "Disease," "disorder," and "condition" are used interchangeably herein.

[0035] Generally, the "effective amount" of a compound refers to an amount sufficient to cause a target biological response. As will be appreciated by those of ordinary skill in the art, the effective amount of the compounds of the invention may vary depending on factors such as the biological target, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and symptoms of the subject. An effective amount includes a therapeutically effective amount and a prophylactically effective amount.

[0036] "Combination" and related terms refer to the simultaneous or sequential administration of the compounds of the invention and other therapeutic agents. For example, the compounds of the invention can be administered simultaneously or sequentially with the other therapeutic agents in separate unit dosage forms, or can be administered simultaneously with the other therapeutic agents in a single unit dosage form. DETAILED DESCRIPTION

[0037] The present invention provides a compound of the following formula (I), or a pharmaceutically acceptable salt, isotopic variant, tautomer or stereoisomer thereof:

[0038] (I) in, Z is selected from O, S, SO2, CH2CH2, CH2 or NH; Z1, Z2, Z3 are each independently selected from O, S or CH2; X1 is selected from CH or N; Y is selected from CH, CF, CMe or N; R is selected from -N(R a )(R a '),OR a , C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-12 membered heterocyclic group, or ; R1 is selected from H, C 1-6 Alkyl or 4-12 membered heterocyclic group; R and R1 are independently replaced by 1-3 R x replace; R x Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkenyl, halogen, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Alkoxy, -(CH2) p CN, NH2, OH, -O(CH2) p OR a 、-O(CH2) p N(R a )(R a '),-N(R a )-(CH2) p OR a 、-O(CH2) p -R a 、-C(O)R a 、-C(O)OR a 、-(CH2) p -OR a 、-SR a 、-S(O)2-R a 、-(CH2) p -C(O)OR a 、-(CH2) p-OC(O)N(R a )(R a '),-N(R a )-S(O)2R a ', -N(R a )-S(O)R a '、-OC(O)N(R a )(R a '); R3 is selected from -NR m R n or a 4-12 membered heterocyclic group; R m Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, CN, -LC(O)R a 、-LC(O)OR a 、-(CH2) p -OR a 、-S(O)2-R a , -L-5-12 membered heteroaryl, -L-phenyl, -L-5-6 membered heterocyclic group; the R m Optionally 1-6 halogens, NH2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, CN, -L-OR a 、-L-SR a 、-LN(R a )(R a '), -NH-(CH2) p -C(O)OR a , C 3-6 further substituted by cycloalkyl, 4-6 membered heterocyclyl or 5-12 membered heteroaryl; R n Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl or C 3-6 Cycloalkyl; When R m When selected from -L-5-12 membered heteroaryl, the 5-12 membered heteroaryl may be further substituted by 1-3 R p replace; R p Selected from halogen, NH2, OH, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C1-6 Haloalkyl, C 1-6 Haloalkoxy or CN; L is selected from a bond, C 1-6 Alkyl or C 1-6 L may be optionally substituted by halogen, OH or C 1-6 Alkoxy substitution; R a Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl, 4-6 membered heterocyclic group, C 3-6 Cycloalkyl or 5-6 membered heteroaryl; R a 'Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl 4-6 membered heterocyclic group or C 3-6 Cycloalkyl; p is independently selected from 0, 1, 2, 3, 4 or 5.

[0039] In another more specific embodiment, the present invention provides a compound of the following formula (II), or a pharmaceutically acceptable salt, isotopic variant, tautomer or stereoisomer thereof: (II) in, X1 is selected from CH or N; Y is selected from CH, CF, CMe or N; R is selected from -N(R a )(R a '),OR a , C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-12 membered heterocyclic group, or ; R1 is selected from H, C 1-6 Alkyl or 4-12 membered heterocyclic group; R and R1 are independently replaced by 1-3 R x replace; R x Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkenyl, halogen, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Alkoxy, -(CH2) p CN, NH2, OH, -O(CH2) p ORa 、-O(CH2) p N(R a )(R a '),-N(R a )-(CH2) p OR a 、-O(CH2) p -R a 、-C(O)R a 、-C(O)OR a 、-(CH2) p -OR a 、-SR a 、-S(O)2-R a 、-(CH2) p -C(O)OR a 、-(CH2) p -OC(O)N(R a )(R a '),-N(R a )-S(O)2R a ', -N(R a )-S(O)R a '、-OC(O)N(R a )(R a '); R3 is selected from -NR m R n or a 4-12 membered heterocyclic group; R m Select, -L-5-12 membered heteroaryl; R n Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl or C 3-6 Cycloalkyl; When R m When selected from -L-5-12 membered heteroaryl, the 5-12 membered heteroaryl may be further substituted by 1-3 R p replace; R p Selected from halogen, NH2, OH, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or CN; L is selected from a bond, C 1-6 Alkyl or C 1-6 Haloalkyl; R a Selected from H, C 1-6 Alkyl, deuterated C1-6 Alkyl, C 1-6 Haloalkyl, 4-6 membered heterocyclic group, C 3-6 Cycloalkyl or 5-6 membered heteroaryl; R a 'Selected from H, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl 4-6 membered heterocyclic group or C 3-6 Cycloalkyl; p is independently selected from 0, 1, 2, 3, 4 or 5.

[0040] In another more specific embodiment, the present invention provides a compound of formula (III), or a pharmaceutically acceptable salt, isotopic variant, tautomer or stereoisomer thereof: (III) in, R is selected from NH2, N(Me)2, , , or ; R is 1-3 optional R x replace; R x is selected from H, methyl, deuterated methyl, F, trifluoromethyl, trifluoromethoxy, methoxy, NH2 or OH; R3 is selected from , , , , , or ; R p is selected from F, NH2, OH, methyl, trifluoromethoxy, difluoromethoxy or CN.

[0041] In another more specific embodiment, the present invention relates to the following compound or its tautomer, stereoisomer or pharmaceutically acceptable salt, wherein the compound is selected from: , , , , , , , or .

[0042] The compounds of the present invention may include one or more asymmetric centers and may therefore exist in a variety of stereoisomeric forms, for example, enantiomers and / or diastereoisomeric forms. For example, the compounds of the present invention may be individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.

[0043] The compounds of the present invention may also exist as tautomers. For compounds that exist in different tautomeric forms, a compound is not limited to any specific tautomer, but is intended to encompass all tautomeric forms.

[0044] The present invention also includes isotopically labeled compounds (isotopic variants) which are identical to those described by formula (A) but in which one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number commonly found in nature. Examples of isotopes that can be introduced into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, for example 2 H. 3 H. 13 C. 11 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 Cl. Compounds of the present invention containing the above isotopes and / or other isotopes of other atoms, their prodrugs and pharmaceutically acceptable salts of the compounds or prodrugs are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as those in which a radioactive isotope (e.g. 3 H and 14 C) can be used for drug and / or substrate tissue distribution assays. 3 H and carbon-14, i.e. 14 C isotopes are particularly preferred because they are easy to prepare and detect. 2H, because higher metabolic stability can provide therapeutic benefits, such as prolonged half-life in vivo or reduced dosage requirements, and thus may be preferred in some cases. Isotope-labeled compounds of formula (A) of the present invention and their prodrugs can generally be prepared by replacing non-isotope-labeled reagents with readily available isotope-labeled reagents when carrying out the processes disclosed in the following schemes and / or Examples and Preparation Examples. Example

[0045] The reagents used in the present invention are commercial reagents purchased directly or synthesized by common methods well known in the art.

[0046] Table 1 Notes on abbreviations used in this application

[0047] Example 1: Preparation of intermediates

[0048]

[0049] Step 1: At 10°C, under nitrogen protection, raw material a1-1 (20.0 g, 117.5 mmol) and raw material allyl acetate a1-2 (15.3 g, 152.7 mmol) were dissolved in a mixed solution of 51 mL toluene and water (v / v, 50 / 1), tetramethylguanidine (27.1 g, 235.0 mmol) was added, and the mixed solution was stirred at 10°C for 30 minutes. Catalyst allyl palladium chloride dimer (30 mg, 0.08 mmol) and (S,S)-diaminocyclohexane-phenyltrost ligand (130 mg, 0.19 mmol) were added to the reaction solution, and the reaction was continued at this temperature for 12 hours, and the reaction was stopped. 250 mL of saturated ammonium chloride aqueous solution was added to the mixed solution, extracted with methyl tert-butyl ether, dried over anhydrous sodium sulfate, and concentrated to obtain compound a1-3 (16.6 g), with a yield of 67%.

[0050] Step 2: Under nitrogen protection at 10°C, the intermediate a1-3 (16.2 g, 77.0 mmol) was dissolved in 65 mL of ethylene glycol, and trimethylsilyl chloride (20.9 g, 192.6 mmol) was added dropwise, and the mixture was reacted at 10°C for 12 hours. Sodium hydroxide aqueous solution (65 mL, 2N) was added to the reaction solution, extracted with methyl tert-butyl ether, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography (petroleum ether / ethyl acetate, 9 / 1) to obtain a yellow oil a1-4 (16.0 g) with a yield of 82%.

[0051] Step 3: In an ice bath, under nitrogen protection, the intermediate a1-4 (16.0 g, 62.9 mmol) and the raw material of the previous step were dissolved in 25 mL of anhydrous tetrahydrofuran, and 9-boranebicyclo[3,3,1]-nonane (0.5 M in tetrahydrofuran) (121 mL, 75.5 mmol) was added dropwise. The mixture was reacted for 1 hour in an ice bath, and methyl chloroacetate a1-5 (8.9 g, 81.78 mmol) was added to the reaction solution. The temperature was lowered to -45°C, and lithium bis(trimethylsilyl)amide (1M, 207 mL) was added dropwise. After the dropwise addition, the reaction was allowed to react at room temperature for 12 hours. Sodium hydroxide aqueous solution (40 mL, 2N) and 50 mL of ethanol were added to the reaction solution, and the temperature was raised to 70°C and the reaction was allowed to react for 8 hours, and the reaction was stopped. 400 mL of ice water was added to the reaction solution, extracted with methyl tert-butyl ether, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography (petroleum ether / ethyl acetate, 3 / 1) to obtain a yellow oil a1 (12.3 g), with a yield of 87%. LCMS ESI-MS m / z: 225 [M+H] + .

[0052] Preparation of intermediate a26

[0053] Step 1: In an ice bath, under nitrogen protection, the intermediate a1 (50.0 g, 222.9 mmol) was dissolved in 500 mL of anhydrous tetrahydrofuran, and lithium bis(trimethylsilyl)amide (1 M in tetrahydrofuran) (245 mL) was added dropwise. After the addition was completed, the mixture was stirred for 1 hour in an ice bath. Diethyl oxalate (39.1 g, 267.5 mmol) was added dropwise to the reaction solution, and the temperature was raised to room temperature for reaction for 2 hours to obtain a mixture. Acetyl chloride (39.3 g, 501.6 mmol) was dissolved in 200 mL of ethanol, and the mixture was stirred for 1 hour in an ice bath to prepare reaction solution S1. Reaction solution S1 and hydroxylamine hydrochloride (16.2 g, 234.1 mmol) were added to the mixture, and the temperature was raised to 65°C and the reaction was continued for 12 hours. The reaction was stopped and the solvent was evaporated under reduced pressure. 500 mL of ice water was added to the reaction solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The mixture was dissolved again in 100 mL of 1,4-dioxane, and the pH was adjusted to about 2 with dilute hydrochloric acid (1M). The mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated to obtain a white solid a26-1 (21.5 g). LCMS ESI-MSm / z: 278 [M+H] + .

[0054] Step 2: Dissolve the intermediate a26-1 (21.5 g, 77.5 mmol) and ammonia (129 mL) in 44 mL of ethanol. The mixture is reacted at room temperature for 12 hours and the reaction is stopped. Add 200 mL of water to the reaction solution, extract with ethyl acetate, dry with anhydrous sodium sulfate, and concentrate to obtain a yellow solid a26-2 (18.0 g). LCMS ESI-MS m / z: 249 [M+H] + .

[0055] Step 3: Dissolve the intermediate mixture a26-2 (18.0 g, 72.5 mmol) and pyridine (13.7 g, 174 mmol) in 72 mL acetonitrile, add trifluoroacetic anhydride (18.2 g, 87.0 mmol) dropwise, and react the mixture at room temperature for 0.5 hours to stop the reaction. Add 400 mL of ice water to the reaction solution, extract with ethyl acetate, dry with anhydrous sodium sulfate, and concentrate to obtain a yellow oil a26 (15.0 g). The total yield of the three steps is 29%. LCMS ESI-MS m / z: 231 [M+H] + .

[0056] Preparation of intermediates a27-a29

[0058] Step 1: In an ice bath, under nitrogen protection, the intermediate a26 (15.0 g, 65.1 mmol) and sodium methoxide (2.93 g, 16.3 mmol) were dissolved in 40 mL of anhydrous methanol and reacted at room temperature for 2 hours. Ammonium chloride (3.8 g, 71.65 mmol, 30 ° C) was added to the reaction solution, and the reaction was continued for 12 hours. The reaction was stopped and the solvent was evaporated under reduced pressure. 500 mL of ice water was added to the reaction solution, and the pH was adjusted to about 8 with saturated sodium bicarbonate aqueous solution. The reaction solution was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. A white solid a27 (14.0 g) was obtained with a yield of 87%. LCMS ESI-MS m / z: 248 [M+H] + .

[0059] Step 2: Dissolve the intermediate 27 (4.0 g, 16.2 mmol) and the raw material a28-1 (8.65 g, 48.6 mmol) in 44 mL N,N-dimethylformamide, add 1,8-diazabicycloundec-7-ene (7.39 g, 48.5mmol), and heat the mixture to 90°C for 12 hours to stop the reaction. Add 200 mL of water to the reaction solution, extract with ethyl acetate, dry over anhydrous sodium sulfate, and concentrate. The crude product was separated by flash reverse column chromatography (chromatographic column: C18; acetonitrile / water = 1 / 1) to obtain a white solid a28-2 (3.3 g) with a yield of 61%. LCMS ESI-MS m / z: 334 [M+H] + .

[0060] Step 3: Dissolve the intermediate a28-2 (3.0 g, 9.0 mmol) and phosphorus oxychloride (9.0 ML) in 18 mL N,N-diethylaniline, heat the mixture to 80°C and react for 12 hours to stop the reaction. Add 100 mL of ice water to the reaction solution, extract with ethyl acetate, dry with anhydrous sodium sulfate, and concentrate. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 4 / 1) to obtain a white solid a28 (1.6 g) with a yield of 48%. LCMS ESI-MS m / z: 370 [M+H] + .

[0061] Referring to the synthetic route of intermediate a28, intermediate a29 was synthesized using similar raw materials / compounds. , LCMS ESI-MS m / z: 352 [M+H] + .

[0062] Synthesis of intermediate b1

[0063] Step 1: In an ice bath, dissolve the intermediate a29 (500 mg, 1.42 mmol) and diisopropylethylamine (1.8 g, 14.2 mmol) in 5 mL of acetonitrile, slowly add the raw material b1-1 (258 mg, 1.70 mmol) to the system, react at room temperature for 1 hour, and stop the reaction. Add 30 mL of ice water to the system, adjust the pH to about 7 with dilute hydrochloric acid, extract with dichloromethane, dry with anhydrous sodium sulfate, and concentrate. The crude product was separated by reverse column chromatography (chromatographic column: C18; acetonitrile / water (10 mmol / L ammonium bicarbonate, 4 / 1) to obtain a yellow solid b1 (550 mg), yield: 90%. LCMS ESI-MS m / z: 431 [M+H] + .

[0064] Synthesis of intermediate c1

[0065] Step 1: In an ice bath, dissolve the raw material c1-1 (4.0 g, 19.32 mmol) and diisopropylethylamine (10.0 g, 77.29 mmol) in 80 mL of acetonitrile, slowly add the intermediate a1 (4.8 g, 21.26 mmol) and magnesium bromide ether (15.0 g, 57.97 mmol) to the system, add the mixture dropwise, heat to 80°C and react for 48 hours, then stop the reaction. Add 300 mL of ice water to the system, adjust the pH to about 5 with dilute hydrochloric acid, extract with dichloromethane, dry with anhydrous sodium sulfate, and concentrate. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 10 / 1) to obtain a yellow oil c1 (3.0 g), with a yield of 39%. LCMS ESI-MS m / z: 399 [M+H] + .

[0066] Synthesis of intermediates C12-C17

[0067] Step 1: Dissolve the raw material c12-1 (3.0 g, 20.8 mmol) and oxalyl chloride (COCl)2 (3.9 g, 31.2 mmol) in 15 mL of dichloromethane under an ice bath, slowly add dimethylformamide (5 drops) to the system, react at room temperature for 2 hours, stop the reaction, and evaporate the solvent under reduced pressure. Dissolve the mixture in 5 mL of dichloromethane to prepare reagent S1. Dissolve the raw material (R)-3-fluoro-tetrahydropyrrole hydrochloride c12-2 (2.8 g, 22.9 mmol) and triethylamine (6.3 g, 62.4 mmol) in 10 mL of anhydrous dichloromethane, add reagent S1 dropwise, add dropwise, react at room temperature for 3 hours, and stop the reaction. 30 mL of ice water was added to the reaction solution, extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography (petroleum ether / ethyl acetate, 1 / 2) to obtain a yellow oil c12-3 (3.5 g), yield: 78%. LCMS ESI-MS m / z: 216 [M+H] + .

[0068] Step 2: In an ice bath, under nitrogen protection, the intermediate c12-3 (3.0 g, 13.9 mmol) of the previous step was dissolved in 20 mL of anhydrous tetrahydrofuran, and lithium aluminum hydride (1.3 g, 34.84 mmol) was added. Stirring was continued for 2 hours in an ice bath to stop the reaction. The reaction solution was poured into 100 mL of ice water, extracted with ethyl acetate, washed with saturated brine, concentrated, and the crude product was separated by column chromatography (100% ethyl acetate) to obtain oily compound c12 (1.3 g) with a yield of 54. LCMS ESI-MS m / z: 174 [M+H] + .

[0069] Referring to the synthetic route of intermediate c12, similar raw materials / compounds were used to synthesize the following intermediates.

[0070] Table 2 Structures and LC-MS of intermediates c13~c17

[0071] Synthesis of intermediate d1-d2

[0072] Step 1: Dissolve intermediate b1 (550 mg, 1.28 mmol), triethylamine (650 mg, 6.38 mmol) and catalyst 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (Pd(dppf)Cl2) (232.2 mg, 1.53 mmol) in 11 mL of methanol. The mixture is heated to 120°C under carbon monoxide (30 atm) for 24 hours, the reaction is stopped, and filtered. Add 100 mL of ice water to the reaction solution, extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate, and separate the crude product by reverse column chromatography (chromatographic column: C18; acetonitrile / water (10 mmol / L ammonium bicarbonate, 5 / 1) to obtain a light yellow solid d1 (400 mg): LCMS ESI-MS m / z: 455 [M+H] + , and d2 (65 mg): LCMS ESI-MS m / z: 441 [M+H] + The yields were 69% and 11.5% respectively.

[0073] Synthesis of intermediate d3

[0074] Step 1: Dissolve the intermediate d2 (65 mg, 1.15 mmol), diisopropylethylamine (96 mg, 0.74 mmol) and the raw material d3-1 (41.2 mg, 0.22 mmol) in 1 mL of dichloromethane, add the condensation reagent 1-hydroxybenzotriazole (20 mg, 0.15 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (42.4 mg, 0.22mmol), and react the mixture at room temperature for 12 hours, stop the reaction, and filter. Add 10 mL of ice water to the reaction solution, extract with dichloromethane, dry with anhydrous sodium sulfate, concentrate, and separate the crude product by reverse column chromatography (chromatographic column: C18; acetonitrile / water (10 mmol / L ammonium bicarbonate, 4 / 1) to obtain a light yellow solid d3 (22 mg), yield: 24%, LCMS ESI-MS m / z: 609 [M+H] + .

[0075] Example 2: Synthesis of target molecule G1

[0076] Step 1: Under nitrogen protection, the intermediate d1 (100 mg, 0.22 mmol), ammonium acetate (30 mg, 0.35mmol) and elemental sulfur (11.3 mg, 0.35 mmol) were dissolved in 1 mL of ethanol, and the mixture was reacted at 60°C for 0.5 hours. Malononitrile (24 mg, 0.36 mmol) was added to the reaction solution, and the temperature was raised to 80°C for 1 hour to stop the reaction. 5 mL of ice water was added to the reaction solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash reverse column chromatography (chromatographic column: C18; acetonitrile / water (10 mmol / L ammonium bicarbonate), 5 / 1) to obtain a light yellow solid G1-1 (105 mg), yield: 89%. LCMS ESI-MS m / z: 535 [M+H] + .

[0077] Step 2: Dissolve the compound G1-1 (40 mg, 0.08 mmol) and ammonia water (0.35 mL) in 0.5 mL of ethanol, react at room temperature for 13 hours, and stop the reaction. Add 10 mL of ice water to the reaction solution, extract with dichloromethane, dry with anhydrous sodium sulfate, and concentrate. The crude product was separated by HPLC preparative chromatography (chromatographic column: XBridge Shield RP18 OBD Column30*150 mm, 5 μm; mobile phase A: water (10 mmol ammonium bicarbonate + 0.05% ammonia water), mobile phase B: acetonitrile; flow rate: 60 mL / min) to obtain a white solid G1 (9.7 mg), yield: 25%. LCMS ESI-MS m / z: 520 [M+H] + .

[0078] 1 H NMR (300 MHz, DMSO-d6,ppm): δ 7.89 (s, 1H), 7.78 (s, 1H), 7.32 (s,1H), 6.96 (s, 2H), 4.56 (s, 1H), 3.74 (s, 1H), 3.20 (s, 1H), 3.09 (d,J= 16.0Hz, 1H), 2.55 (d,J= 4.9 Hz, 3H), 1.99 (dd,J= 20.9, 9.7 Hz, 13H), 1.15 (s,3H).

[0079] Example 3: Synthesis of target molecules G2-G3

[0080] Step 1: Under nitrogen protection, the intermediate d3 (22 mg, 0.04 mmol), ammonium acetate (4.5 mg, 0.06 mmol) and elemental sulfur (1.85 mg, 0.06 mmol) were dissolved in 0.5 mL of ethanol, and the mixture was reacted at 60°C for 0.5 hours. Malononitrile (4.0 mg, 0.06 mmol) was added to the reaction solution, and the temperature was raised to 80°C for 1 hour to stop the reaction. 5 mL of ice water was added to the reaction solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a yellow solid G2-1 (24 mg), with a yield of 96%. LCMS ESI-MS m / z: 689 [M+H] + .

[0081] Step 2: Dissolve the compound G2-1 (24 mg, 0.04 mmol) in 0.5 mL of dichloromethane, add a solution of hydrogen chloride in 1,4-dioxane (0.5 mL, 4M), react at room temperature for 1 hour, stop the reaction, and remove the solvent under reduced pressure. The crude product was separated by HPLC preparative chromatography (chromatographic column: XBridge Shield RP18 OBD Column 30*150 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min) to obtain a white solid G2 (4.8 mg), yield: 22%. LCMS ESI-MS m / z: 589 [M+H] + .

[0082] 1 H NMR (300 MHz, DMSO-d6, formate): δ 8.34 (s, 1H), 6.93 (d, J = 8.7 Hz, 3H), 3.73 (s, 2H), 3.56 (s, 2H), 3.31 (s, 3H), 3.02 (d, J = 15.6 Hz, 1H), 2.77(s, 2H), 2.67 (s, 2H), 2.48 (s, 4H), 2.05 – 1.70 (m, 9H), 1.65-1.42 (m, 3H),1.14 (s, 3H).

[0083] Referring to the synthetic route of target molecule G2, similar raw materials / intermediates (such as intermediates) are used. etc.), synthesize target molecule synthesize target molecule G3 , LC-MS [M+H] + It is 659.

[0084] 1H NMR (300 MHz, DMSO-d6, ppm): δ 8.45 (s, 1H), 7.26 (s, 1H), 6.93 (s,2H), 4.52 (s, 1H), 3.71 (s, 1H), 3.55 (t, J = 4.6 Hz, 4H), 3.42 (s, 1H), 3.33(s, 3H), 3.05 (d, J = 15.9 Hz, 1H), 2.72 (s, 1H), 2.53 (s, 2H), 2.45 (s, 4H),2.04 – 1.89 (m, 9H), 1.84 (s, 3H), 1.22 (s, 1H), 1.12 (s, 3H), 1.00 (s, 2H),0.69 (s, 2H).

[0085] Embodiment 4: Compounds for KRAS G12D Mediated p-ERK inhibition test (directly reflects the cellular level inhibitory effect of the test compound). Details are as follows: AGS cells cultured in F-12K medium (Gibco, Cat. No. 30-2004) containing 10% fetal bovine serum and 1% penicillin-streptomycin were seeded in 384-well microplates and incubated overnight at 37°C with 5% CO2. 200 nL of different concentrations of compound (1000 nM starting concentration, 4-fold dilution) were added to each well and incubated at 37°C for 3 hours. Then, cells were fixed in 8% fixative (Solarbio, Cat. No. P1112) and washed once with phosphate-buffered saline (PBS). After washing, blocking solution (LI-COR, Cat. No. 927-40000) was added to each well and blocked for 1 hour at room temperature. After removing the blocking solution, phospho-p44 / 42 MAPK(T202 / Y204) Rabbit mAb (Cell Signaling Technology (CST), Cat. No. 4370S) and GAPDH(D4C6R) Mouse mAb (Cell Signaling Technology (CST), Cat. No. 97166S) antibody working solutions were added to each well and incubated overnight at 4°C. The microplate was washed three times with phosphate-buffered saline (PBST) containing 0.1% Tween-80, and IRDye 800CW Goat anti-Rabbit IgG (H+L) (LI-COR, Cat. No. 926-32211) and IRDye 680RD Goat anti-Mouse IgG (H+L) (LI-COR, Cat. No. 926-68070) antibody working solutions were added, and the microplate was incubated at room temperature in the dark. After washing the microplate three times with phosphate buffered saline (PBST), the microplate was centrifuged at 1000 rpm for 1 minute, and the plate was scanned and read using an Odyssey CLx (LI-COR) instrument to record the signal value.

[0086] IC 50 The calculation formula Calculate compound IC using nonlinear regression equation 50 Value: Y = Lower platform signal + (Upper platform signal - Lower platform signal) / (1+10^ ((LogIC 50 -X)*Hill slope)); X = logarithm of compound concentration.

[0087] Table 3 Target molecules for AGS (KRAS G12D pERK inhibition effect on ) cells

[0088]

[0089] After testing, the molecule of the present invention has an effect on KRAS G12DMutated tumor cells have a good inhibitory effect and are expected to be able to inhibit KRAS G12D To achieve better tumor suppression effect.

[0090] Embodiment 5: Compounds for KRAS G12V Mediated p-ERK inhibition test (directly reflects the cellular level inhibitory effect of the test compound). Details are as follows: SW480 cells cultured in DMEM medium (Gibco, Cat. No. 11995065) containing 10% fetal bovine serum and 1% penicillin-streptomycin were seeded in 384-well microplates and incubated overnight at 37°C with 5% CO2. 200 nL of different concentrations of compound (1000 nM starting concentration, 4-fold dilution) were added to each well and incubated at 37°C for 3 hours. Then, cells were fixed in 8% fixative (Solarbio, Cat. No. P1112) and washed once with phosphate-buffered saline (PBS). After washing, blocking solution (LI-COR, Cat. No. 927-40000) was added to each well and blocked for 1 hour at room temperature. After removing the blocking solution, the antibody working solution recognizing phospho-p44 / 42 MAPK(T202 / Y204) phospho-p44 / 42 MAPK(T202 / Y204) Rabbit mAb (Cell Signaling Technology (CST), Cat. No. 4370S) and GAPDH(D4C6R) Mouse mAb (CST, Cat. No. 97166S) was added to each well and incubated at 4°C overnight. The microplate was washed three times with PBS solution containing 0.1% Tween-80 (PBST), and IRDye 800CW Goat anti-Rabbit IgG (H+L) (LI-COR, catalog number 926-32211) and IRDye 680RD Goat antiMouse IgG (H+L) (LI-COR, catalog number 926-68070) antibody working solution were added, and the microplate was incubated at room temperature in the dark. After washing the microplate three times with PBST, the microplate was centrifuged at 1000 rpm for 1 minute, and the plate was scanned and read using the Odyssey CLx (LI-COR) instrument to record the signal value.

[0091] IC 50 The calculation formula Calculate compound IC using nonlinear regression equation 50 Value: Y = Lower platform signal + (Upper platform signal - Lower platform signal) / (1+10^ ((LogIC 50-X)*Hill slope)); X = logarithm of compound concentration.

[0092] Target molecule G3 for SW480 (KRAS G12v ) cells with an IC 50 =44nM.

[0093] After testing, the molecule of the present invention has an effect on KRAS G12V Mutated tumor cells have a good inhibitory effect and are expected to be able to inhibit KRAS G12V To achieve better tumor suppression effect.

[0094] Embodiment 6:

[0095] 3D antiproliferative effects of the compounds on KRAS G12D mutant pancreatic cancer HPAC cell lines.

[0096] Cell culture: HPAC pancreatic cancer cells were cultured in RPMI 1640 medium (Gibco, Cat. No. A10491-01) containing 10% fetal bovine serum and 1% penicillin / streptomycin in T75 culture flasks (Corning, Cat. No. 430641).

[0097] Cell culture: SW620 colorectal cancer cells were cultured in RPMI 1640 medium (Gibco, Cat. No. A10491-01) containing 10% fetal bovine serum and 1% penicillin / streptomycin in T75 culture flasks (Corning, Cat. No. 430641).

[0098] Experimental process: Use a nanoliter pipetting system to add 200nL (1000 nM, 4-fold dilution) of the diluted test compound to a 384-well low-adsorption cell culture plate. After the cells are plated, place the culture plate in a 37°C, 5% CO2 constant temperature incubator for 3D culture. After the test compound and cells are incubated for 5 days, add CellTiter-Glo® 3D reagent (Promega, category number G9683) to each well, and use an Envision multi-function microplate reader (Perkin Elmer, category number Envision 2104) to read the luminescence value. The light signal is proportional to the amount of ATP in the system, and the ATP content directly represents the number of live cells in the system. Finally, the IC value of the compound is obtained using the nonlinear fitting formula using the XLFIT software. 50 (half inhibitory concentration).

[0099] Inhibition rate (%) = 100 × (negative control average value - compound reading value) / (negative control average value - positive control average value), negative control: DMSO. Positive control: culture medium.

[0100] After testing, it was found that the molecule G3 of the present invention has a good inhibitory effect on KRAS mutant cell lines.

Claims

1. A spirocyclic compound, characterized in that: Has the following structure: , , , , , , , or .

2. A use of the spirocyclic compound as claimed in claim 1, characterized in that: The invention relates to a use of the compound as an active ingredient in preparing a drug for treating and / or preventing a disease mediated by a KRAS mutant.

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