Tricyclic compounds as kras g12c inhibitors and uses thereof

Abstract

The present invention provides a novel tricyclic compound represented by formula (I) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the same, and use thereof in the preparation of a drug for preventing and / or treating KRAS G12C mutant tumors.

Description

[0001] The present application claims priority to the prior application with the patent application number of 202110162783.3, the title of "Tricyclic Compounds as KRAS G12C Inhibitors and Uses Thereof", which was filed on February 5, 2021 with the State Intellectual Property Office of China. The entire contents of the above-mentioned prior application are incorporated herein by reference. TECHNICAL FIELD

[0002] The present application relates to a novel tricyclic compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the same and the use as KRAS G12C inhibitors in the prevention or treatment of related diseases. BACKGROUND

[0003] Ras is the first discovered human proto-oncogene, which has three family members: Hras, Kras and Nras (Barbacid M, Annu Rev Biochem, 1987; 56: 779-827). RAS as a class of small G proteins has GTP hydrolysis enzyme activity, is located in the inner side of the plasma membrane, and regulates its activity by different binding with GTP / GDP. When it binds with GTP, it is activated (on), and when it binds with GDP, it is in an inactivated state (off). Ras GTP kinase is essential in many signal networks, which has the function of signal integration and transmits signals to downstream effectors, participates in life activities such as cell movement, cytoskeleton assembly, vesicle and nuclear transport, and further regulates life processes such as cell proliferation, differentiation, aging and apoptosis (Fernandez-Medarde A, et al, Genes Cancer, 2011; 2(3): 344-58). Therefore, RAS protein is considered as an important molecular switch protein in cell signal transmission.

[0004] Once the proto-oncogene Ras is activated, it becomes an oncogene with carcinogenic activity. There are three ways to activate the Ras gene: point mutation, overexpression, and insertion activation. Among them, the most common way of activating the Ras gene is point mutation, and the carcinogenic effect is mainly through point mutation. At present, 151 different Ras point mutations have been found, mainly concentrated in the mutations of 12, 13 glycine and 61 glutamine (Prior IA, et al, Cancer Res 2012; 72(10): 2457-67). Among the above mutations, G12 point mutation is the most common, and G12 mutation is dominant in KRAS and Hras. In KRAS mutation, 15 different point mutations of G12 have been found, including G12A, G12D, G12F, G12K, G12N, G12S, G12V, G12Y, G12C, G12E, G12I, G12L, G12R, G12T and G12W. Among them, G12D mutation accounts for about 41% of all G12 mutations, G12V accounts for about 28%, and G12C accounts for about 14% (Hobbs GA, et al, Cancer Cell, 2016; 29(3): 251-3).

[0005] Ras mutation is an important reason for promoting the occurrence of various cancers, which often occurs in the early stage of tumor occurrence. These activated RAS proteins promote uncontrolled cell growth and proliferation. According to statistics, the five cancers with the highest frequency of Ras mutation are pancreatic ductal adenocarcinoma, colorectal cancer, multiple myeloma, lung cancer and skin melanoma, with mutation frequencies of 97.7%, 52.2%, 42.6%, 32.2% and 29.1% respectively (Albertini AF, et al, Bull Cancer, 2017; 104(7 / 8): 662-74). It is worth noting that among these Ras mutations, the mutation frequency of KRAS is significantly higher than that of the other two mutations. For example, in pancreatic ductal adenocarcinoma, the mutation rate of KRAS is as high as 97.7%, while Nras and Hras are all 0; while in colorectal cancer, 52.2% of the Ras mutation rate is as high as 44.7% in KRAS. In the study of non-small cell lung cancer, it was found that Ras mutation often occurs in codon 12, the most common being G12C point mutation, followed by G12V and G12D (Yoon YK, et al, Mol Carcinog 2010; 49(4): 353-62).

[0006] Activating mutations in the KRAS gene are closely related to the occurrence and development of human malignancies and tumor recurrence. Genetic and biochemical studies have shown that KRAS-dependent signaling plays an important role in regulating the growth, proliferation, invasion and metastasis of various cancer cells. In terms of patient prognosis, KRAS is also considered a marker. In non-small cell lung cancer patients, KRAS mutant patients have a shorter survival time than KRAS wild-type patients, especially patients with G12C point mutations (Svaton M, et al, Anticancer Res, 2016; 36(3): 1077-82). Therefore, the scientific community has been committed to finding small molecules that can bind to specific mutant KRAS proteins and inhibit the activation and function of KRAS proteins, thereby preventing the transduction of downstream signaling pathways, and ultimately playing a role in inhibiting tumor growth.

[0007] In 2013, the Shokat group of the Howard Hughes Medical Institute first reported KRAS G12C small molecule inhibitors in Nature (Ostrem J M, et al, Nature, 2013, 503(7477): 548-551). These inhibitors can bind to the allosteric binding pocket below the switch II region of the KRAS protein molecule and form a covalent bond with the nearby Cys12, thereby selectively inhibiting the activation of KRAS G12C. Based on the above research, a new KRAS G12C irreversible covalent inhibitor ARS-853 was reported by Wellspring in 2016, which can fix KRAS protein in the inactive GDP binding conformation and has selective inhibitory activity on various KRAS G12C mutant tumor cells (Patricelli M P, et al, Cancer Discov, 2016, 6(3): 316-329; Lito P, et al, Science, 2016, 351(6273): 604-608).

[0008] At present, heterocyclic compounds that have been tried as KRAS G12C covalent inhibitors include benzopyrimidinones, pyridopyrimidinones, and tetrahydropyridopyrimidines, etc. (WO2017 / 201161, WO2018 / 119183, WO2018 / 217651, WO2018 / 206539, WO2018 / 143315, WO2017 / 087528, WO2020 / 081282, WO / 2020 / 178282). The above compounds are all described as KRAS inhibitors for the treatment of tumors. Given the huge tumor market and unmet market demand, the development of KRAS G12C inhibitors has great market prospects. SUMMARY

[0009] The present application provides a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0010]

[0011]

[0012] wherein

[0013] R 1 is selected from the following groups optionally substituted with R 1a C6-C 10 aryl, 5-10 membered heteroaryl;

[0014] R 2 , R 3 are independently selected from H, F, Cl, Br, I, CN, OH or the following groups optionally substituted with R 2a NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, C1-C6 alkoxy, C3-C6 cycloalkyloxy, 4-7 membered heterocyclyloxy, C2-C6 alkenyl, C2-C6 alkynyl;

[0015] R 4a , R 4b are selected from H, or R 4a , R 4b together form =0;

[0016] R 5 is selected from

[0017] Q is selected from C(=0), C(=NR 8 ), NR 9 C(=0), S(=0)2 or NR 9 S(=0)2;

[0018] R 8 , R 9 are independently selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, C6-C 10 aryl or 5-10 membered heteroaryl;

[0019] when R 5 is selected from , R 6 , R 7A , R 7B are each independently selected from H, F, Cl, Br, I, CN, carboxyl or the following groups optionally substituted with R 6a C1-C6 alkyl, C3-C 10cycloalkyl, 4-10 membered heterocyclyl, or R 6 7A and the carbon atom to which they are attached together form a partially saturated C5-C 10 cycloalkyl or 5-10 membered heterocyclyl, said partially saturated C5-C 10 cycloalkyl or 5-10 membered heterocyclyl is optionally substituted with R 6a ;

[0020] when R 5 is selected from , R 7C is selected from H or the following groups optionally substituted with R 6a : C1-C6 alkyl, C3-C 10 cycloalkyl, 4-10 membered heterocyclyl;

[0021] R 10 is selected from C1-C3 alkyl;

[0022] n is selected from 0, 1, 2, or 3;

[0023] L is selected from O, NH, CH2, C(=O), S(=O)2, or S(=O);

[0024] R 11 is selected from the following groups optionally substituted with R 11a : C6-C 10 aryl, 5-10 membered heteroaryl;

[0025] each R 1a , R 2a , R 6a , R 11a is independently selected from F, Cl, Br, I, OH, CN, =O, or the following groups optionally substituted with R b : C1-C6 alkyl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, C1-C6 alkoxy, C3-C6 cycloalkyloxy, 4-10 membered heterocyclyloxy, NH(C1-C6 alkyl), S(O)2(C1-C6 alkyl);

[0026] each R b is independently selected from F, Cl, Br, I, OH, CN, =O, NH2, SH, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclyl.

[0027] In some embodiments, R 1 is selected from the following groups optionally substituted with R 1a : phenyl, 5-10 membered heteroaryl.

[0028] In some embodiments, R 1 is selected from the following groups optionally substituted with R​1a substituted phenyl, pyridyl, benzothiazolyl.

[0029] In some embodiments, R 1 is selected from optionally substituted phenyl, pyridyl, benzothiazolyl. 1a substituted phenyl, pyridyl, benzothiazolyl.

[0030] In some embodiments, R 1 is selected from optionally substituted phenyl. 1a substituted phenyl.

[0031] In some embodiments, R 1a is selected from F, CI, Br, I, OH, CN, NH2, =0, or optionally substituted with R b substituted C1-C3 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, C1-C3 alkoxy, C3-C6 cycloalkyloxy, 4-7 membered heterocyclyloxy, NH(C1-C3 alkyl).

[0032] In some embodiments, R 1a is selected from F, CI, Br, I, OH, CN, NH2, or optionally substituted with F C1-C3 alkyl, C1-C3 alkoxy.

[0033] In some embodiments, R 1a is selected from F, CI, NH2, OH, methyl, or trifluoromethyl.

[0034] In some embodiments, R 1 is selected from

[0035] In some embodiments, R 1a is selected from F or OH.

[0036] In some embodiments, R 1 is selected from

[0037] In some embodiments, R 2 , R 3 are independently selected from H, F, CI, Br, I, CN, OH, or optionally substituted with R 2a substituted NH2, C1-C3 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, C1-C3 alkoxy, C2-C3 alkenyl, C2-C3 alkynyl.

[0038] In some embodiments, R 2 , R 3 are independently selected from H, F, CI, Br, I, or optionally substituted with R 2aSubstituted: C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, C2-C3 alkenyl, C2-C3 alkynyl.

[0039] In some embodiments, R 2a Selected from halogen, CN, OH, ═O or C1-C3 alkyl.

[0040] In some embodiments, R 2 is selected from H, halogen, ethynyl, 1-propynyl, cyclopropyl, trifluoromethoxy or difluoromethoxy.

[0041] In some embodiments, R 2 is selected from H or halogen.

[0042] In some embodiments, R 2 Selected from H or F.

[0043] In some embodiments, R 3 Selected from H, halogen, C1-C3 alkyl or C3-C6 cycloalkyl.

[0044] In some embodiments, R 3 is selected from H or halogen.

[0045] In some embodiments, R 3 is selected from H, F or Cl.

[0046] In some embodiments, R 5 Selected from

[0047] In some embodiments, R 5 Selected from

[0048] In some embodiments, Q is selected from C(=O) or S(=O)2.

[0049] In some embodiments, R 6 、R 7A 、R 7B 、R 7C Each independently selected from H, F, Cl, Br, I, CN or optionally replaced by R 6a Substituted groups include: C1-C3 alkyl, C3-C6 cycloalkyl, and 4-7 membered heterocyclic groups.

[0050] In some embodiments, R 6a Selected from halogen, CN, OH, ═O or C1-C3 alkyl.

[0051] In some embodiments, R 6 、R 7A 、R 7B 、R7C Each is independently selected from H, F, Cl, Br, I, CN, C1-C3 alkyl.

[0052] In some embodiments, R 6 、R 7A 、R 7B 、R 7C Each is independently selected from H, F, Cl, Br, and I.

[0053] In some embodiments, R 6 、R 7A 、R 7B 、R 7C Each is independently selected from H and F.

[0054] In some embodiments, R 7A 、R 7B At least one of them is selected from H.

[0055] In some embodiments, R 7A Selected from H, R 7B is selected from H, F, Cl, Br, I, CN or C1-C3 alkyl.

[0056] In some embodiments, R 6 、R 7A 、R 7B 、R 7C are each independently selected from H.

[0057] In some embodiments, R 5 Selected from

[0058] In some embodiments, R 10 Selected from methyl.

[0059] In some embodiments, n is selected from 0.

[0060] In some embodiments, L is selected from O or NH.

[0061] In some embodiments, R 11 Selected from R 11a Substituted: phenyl, 5-10 membered heteroaryl.

[0062] In some embodiments, R 11 Selected from R 11a Substituted: phenyl, 5-6 membered heteroaryl, or 9-10 membered heteroaryl.

[0063] In some embodiments, R 11 Selected from R 11asubstituted phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, thiazolyl, oxazolyl, imidazolyl, isothiazolyl, isoxazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzopyrrolyl, benzofuranyl, benzothienyl, benzopyrazolyl, pyrimido-pyrazolyl, pyridino-pyrazolyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, benzopyridazinyl.

[0064] In some embodiments, R 11 is selected from optionally substituted phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl. 11a is selected from optionally substituted phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl.

[0065] In some embodiments, R 11 is selected from optionally substituted phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl. 11a is selected from optionally substituted phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl.

[0066] In some embodiments, R 11a is selected from F, CI, Br, I, OH, CN, NH2, =0, or optionally substituted C1-C3 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, C1-C6 alkoxy. b is selected from optionally substituted C1-C3 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, C1-C6 alkoxy.

[0067] In some embodiments, R 11a is selected from C1-C3 alkyl.

[0068] In some embodiments, R 11a is selected from methyl or isopropyl.

[0069] In some embodiments, R 11 is selected from

[0070] In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is selected from a compound of Formula (II) or a pharmaceutically acceptable salt thereof:

[0071]

[0072] wherein R 1 , R 2 , R 3 , R 4a , R 4b , R 11 , and L are as defined above.

[0073] In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0074] In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0075]

[0076] In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0077]

[0078] In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0079]

[0080]

[0081] The present application also provides a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0082] Further, the present application relates to the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the manufacture of a medicament for the prevention or treatment of a KRAS G12C related disease.

[0083] Further, the present application relates to the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the prevention or treatment of a KRAS G12C related disease.

[0084] Further, the present application relates to a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the prevention or treatment of a KRAS G12C related disease.

[0085] The present application also relates to a method of treating a KRAS G12C related disease, comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical preparation comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0086] In preferred embodiments of the present application, the KRAS G12C related disease includes, but is not limited to, inflammatory diseases, autoimmune diseases and cancer.

[0087] Terminology definitions and explanations

[0088] Unless otherwise indicated, the definitions of groups and terms in the specification and claims of this application, including its definition of terms, examples of definitions, preferred definitions, definitions set forth in tables, definitions of specific compounds in the examples, and the like, can be combined and applied to one another in any and all technically appropriate combinations. The group definitions and compound structures resulting from such combinations and applications should be within the scope of the present application as recited in the specification.

[0089] The term "pharmaceutically acceptable salt" means a pharmaceutically acceptable non-toxic acid or base salt, including inorganic acids and bases, organic acids and bases.

[0090] The term "stereoisomer" means isomers that have the same molecular formula but different structures resulting from the spatial arrangement of atoms. Stereoisomers include enantiomers and diastereomers.

[0091] The compounds of the present application can have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms (optical centers) or asymmetric double bonds. Racemates, enantiomers, diastereomers, geometric isomers are all included within the scope of the present application.

[0092] The graphical representation of racemic or enantiomerically pure compounds herein is from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise indicated, a wedge and hashed bond ( and ) represent the absolute configuration of a stereogenic center, and a solid and hashed bond ( and ) represents the cis and trans configuration of a cycloalkyl ring. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they include E and Z geometric isomers unless otherwise indicated. Likewise, all tautomeric forms are included within the scope of the present application.

[0093] The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as the racemic mixtures and other mixtures of the enantiomers or diastereomers, all of which are intended to be within the scope of the present application. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms can be present in the substituents of the compounds of this application. All such isomers, as well as mixtures thereof, are included within the scope of the present application. Compounds of the present application having asymmetric atoms can be isolated in optically active or racemic forms. This application also envisions enriched or resolved mixtures of optically active compounds.

[0094] The term "tautomer" refers to functional isomers resulting from the rapid shift of an atom between two positions in a molecule. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist as two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually result in a mixture with physical and chemical properties consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones, such as acetaldehyde, the keto form predominates, while in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

[0095] The term "pharmaceutical composition" refers to a mixture of one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs with other chemical components, such as physiologically / pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration of the compound to an organism.

[0096] The term "substituted" refers to the replacement of any one or more hydrogen atoms on a particular atom by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is an oxo (i.e., =O), it means that two hydrogen atoms are replaced. Oxo does not occur on aromatic groups.

[0097] The term "optionally" or "optionally" means that the event or circumstance described subsequently may or may not occur, and the description includes both the occurrence of the event or circumstance and the non-occurrence of the event or circumstance. For example, an ethyl group is "optionally" substituted with a halogen, meaning that the ethyl group may be unsubstituted (CH2CH3), monosubstituted (such as CH2CH2F), polysubstituted (such as CHFCH2F, CH2CHF2, etc.), or fully substituted (CF2CF3). It will be understood by those skilled in the art that for any group containing one or more substituents, no substitution or substitution pattern that would be sterically impossible and / or incomposable to synthesize will be introduced.

[0098] The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

[0099] The term "C5-C 10 "Cycloalkyl" is understood to be a saturated or partially saturated non-aromatic monovalent cyclic hydrocarbon radical having 5, 6, 7, 8, 9 or 10 carbon atoms.

[0100] The term "C1-C6alkyl" is to be understood as meaning a straight chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms. Said alkyl group is, for example, a methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, t-butyl, isopentyl, 2-methylbutyl, 1 -methylbutyl, 1 -ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1 -dimethyl- propyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1 -methylpentyl, 2- ethylbutyl, 1 -ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1 -dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl group and the like; preferably, "C1-C6alkyl" can comprise "C1-C3alkyl", which is to be understood as meaning a straight chain or branched saturated monovalent hydrocarbon group having 1, 2, 3 carbon atoms.

[0101] The term "alkoxy" can be understood as "alkyl oxy" or "alkyl-O-", preferably, "C1-C6alkoxy" can comprise "C1-C3alkoxy".

[0102] The term "C3-C 10 The term "C3-C 10 The term "C3-C6cycloalkyl" is to be understood as meaning a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms.

[0103] The term "cycloalkyloxy" can be understood as "cycloalkyl-O-", preferably, "C3-C 10 The term "C3-C6cycloalkyloxy" can comprise "C3-C4cycloalkyloxy".

[0104] The term "4-10 membered heterocyclyl" means a saturated or partially saturated, monovalent monocyclic, fused, spiro, or bridged ring containing 1-5, preferably 1-3, heteroatoms selected from N, O, B, and S. In particular, "4-10 membered heterocyclyl" includes "5-10 membered heterocyclyl", "4-7 membered heterocyclyl"; said "4-10 membered heterocyclyl" includes, but is not limited to: 4-membered rings, such as azetidinyl, oxetanyl; 5-membered rings, such as tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 1,3,2-dioxaborolanyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl; or partially saturated 6-membered rings such as tetrahydropyridinyl. Optionally, the heterocyclyl group can be benzo-fused. The heterocyclyl group can be bicyclic, for example, but not limited to, 5,5 membered rings, such as hexahydrocyclopenta[c]pyrrol-2(lH)-yl ring, or 5,6 membered bicyclic rings, such as hexahydropyrrolo[l,2-a]pyrazin-2(lH)-yl ring. The ring containing nitrogen atoms can be partially unsaturated, i.e. it can contain one or more double bonds, for example, but not limited to, 2,5-dihydro-lH-pyrrolyl, 4H-[l,3,4]thiadiazinyl, 4,5-dihydrooxazolyl, or 4H-[l,4]thiazinyl, or it can be benzo-fused, for example, but not limited to, dihydroisoquinolinyl. Optionally, the 4-10 membered heterocyclyl group can be "4-10 membered heterocycloalkyl", meaning a saturated monovalent monocyclic, fused, spiro, or bridged ring containing 1-5 heteroatoms; preferably, "4-10 membered heterocycloalkyl" includes 5-10 membered heterocycloalkyl and 4-7 membered heterocycloalkyl, also 5-6 membered heterocycloalkyl, etc.

[0105] The term "heterocyclyloxy" can be understood as "heterocyclyl-O-".

[0106] The term "C6-C 10 Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic, monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 10 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), for example phenyl; or a ring having 9 carbon atoms ("C9-aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C10-aryl"), for example tetrahydronaphthyl, dihydronaphthyl or naphthyl. 10 Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic, monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 10 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), for example phenyl; or a ring having 9 carbon atoms ("C9-aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C10-aryl"), for example tetrahydronaphthyl, dihydronaphthyl or naphthyl.

[0107] The term "5-10 membered heteroaryl" is to be understood as including monovalent monocyclic, bicyclic aromatic ring systems having 5 to 10 ring atoms and comprising 1 to 5 heteroatoms independently selected from N, O and S, including but not limited to thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl and the like as well as their benzo derivatives, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzoisoxazolyl, benzoimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, piperazinyl, pyrimidinyl, pyrazinyl, triazinyl and the like, as well as their benzo derivatives, such as quinolinyl, quinazolinyl, isoquinolinyl and the like. "5-6 membered heteroaryl" means a monovalent monocyclic aromatic ring system having 5 or 6 ring atoms and comprising 1 to 4, preferably 1 to 3 heteroatoms independently selected from N, O and S. "9-10 membered heteroaryl" means a monovalent bicyclic aromatic ring system having 9 or 10 ring atoms and comprising 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O and S.

[0108] The term "C2-C6alkenyl" is to be understood as meaning straight-chain or branched one valent hydrocarbon radicals, which contain one or more double bonds and have 2, 3, 4, 5 or 6 carbon atoms, examples including but not limited to ethenyl (-CH=CH2), prop-1-enyl (-CH=CHCH3), prop-2-enyl (-CH2CH=CH2), 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, but-1,3-dienyl, 2-methyl-1,3-butadienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl or hex-4-enyl. Preferably, "C2-C6alkenyl" includes "C2-C3alkenyl".

[0109] The term "C2-C6alkynyl" is to be understood as meaning straight-chain or branched one valent hydrocarbon radicals, which contain one or more triple bonds and have 2, 3, 4, 5 or 6 carbon atoms, examples including but not limited to ethynyl (-C≡CH), prop-1-ynyl (1-propynyl, -C≡CCH3), prop-2-ynyl (propargyl), but-1-ynyl, but-2-ynyl or but-3-ynyl. Examples of "C2-C3alkynyl" include ethynyl (-C≡CH), prop-1-ynyl (1-propynyl, -C≡CCH3), prop-2-ynyl (propargyl).

[0110] In particular, the term "treatment" means that the compounds or formulations described herein are administered to prevent, ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0111] (i) preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed to the disease but has not yet been diagnosed as having it;

[0112] (ii) inhibiting the disease or condition, i.e., arresting its development;

[0113] (iii) relieving the disease or condition, i.e., causing regression of the disease or condition.

[0114] The term "therapeutically effective amount" means an amount of a compound of the present application that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein. The amount of a compound of the present application that will constitute a "therapeutically effective amount" will depend on the compound, the disease state being treated, the severity or the disease state, the age of the mammal, and the manner of administration, but can be determined routinely by the skilled practitioner without undue experimentation.

[0115] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of classes of excipients include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients can enhance the handling properties of a pharmaceutical formulation, i.e., make the formulation more amenable to direct compression by increasing flow and / or cohesion. Examples of typical "pharmaceutically acceptable carriers" suitable for use in the above formulations are: sugars, starches, celluloses and derivatives thereof, and the like excipients commonly used in pharmaceutical formulations.

[0116] The term "pharmaceutically acceptable excipient" refers to those excipients that do not stimulate an undesirable response in an organism and do not substantially abrogate a desired biological activity of an active compound. Suitable excipients are well known to those skilled in the art, e.g., carbohydrates, waxes, water soluble and / or swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0117] The words "comprise," "comprising," or "comprises" and the like are to be construed in an open, non-exclusive sense, i.e., meaning "including but not limited to."

[0118] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by a combination of the embodiments set forth below with other chemical synthetic methods well known in the art, and equivalents thereof as appreciated by those skilled in the art, preferred embodiments including but not limited to the examples of the present application.

[0119] The present application also includes isotopically labeled compounds of the present application that are identical to those described herein, but in which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0120] Certain isotope-labeled compounds of the present application (e.g. 3 H and 14 C-labeled) can be used in compound and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron emitting isotopes, such as 15 O. 13 N. 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present application can generally be prepared by following procedures analogous to those disclosed in the Schemes and / or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0121] In addition, the use of heavier isotopes such as deuterium (i.e. 2 H)) substitution may offer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and may therefore be preferred in certain circumstances, wherein deuterium substitution may be partial or complete, partial deuterium substitution meaning that at least one hydrogen is replaced by at least one deuterium.

[0122] The pharmaceutical compositions of the present application can be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.

[0123] Typical routes of administering the compounds of the present application, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, include, but are not limited to, oral, rectal, topical, inhalant, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

[0124] The pharmaceutical compositions of the present application can be manufactured in a manner appropriate to the type of composition by known methods, such as the conventional methods used for mixing, dissolving, granulating, dragee-making, levigating, emulsifying, freeze-drying or lyophilizing processes.

[0125] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients can enable the compounds of the present application to be formulated as tablets, pills, dragees, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to a patient.

[0126] Solid oral compositions can be prepared by conventional mixing or compaction methods. For example, the active compound can be mixed with a solid excipient, optionally ground, and if necessary, with other suitable excipients, and then processed to form a granulation, which is again processed to form a tablet or dragee core. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, sweeteners, or flavoring agents.

[0127] The pharmaceutical composition can also be adapted for parenteral administration, such as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.

[0128] In all methods of administration of the compounds of the general formula I described herein, the daily dose is from 0.01 to 100 mg / kg of body weight, preferably from 0.05 to 50 mg / kg of body weight, and more preferably from 0.1 to 30 mg / kg of body weight, in single or divided doses.

[0129] The chemical reactions of the specific embodiments are performed in solvents appropriate to the reagents and materials employed and suitable for the chemical changes being effected. For general procedures, modifications of the procedures described in the examples can be necessary in order to achieve the compounds of the present application. DETAILED DESCRIPTION

[0130] The following examples illustrate the technical solutions of the present application, but the scope of protection of the present application includes but is not limited to this.

[0131] Unless otherwise stated, the proportions indicated for mixtures of solvents are volume proportions.

[0132] Unless otherwise indicated, % means percent by weight wt%.

[0133] Compounds were prepared by hand or Software nomenclature, commercially available compounds used the supplier catalog name.

[0134] The structure of the compounds was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The units of NMR shifts are 10 -6 (ppm). The solvent for NMR determination was deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard was tetramethylsilane (TMS); "IC 50 " refers to the half maximal inhibitory concentration, the concentration at which an action is inhibited by half of its maximal inhibition. The eluent in the following can be a mixed eluent formed by two or more solvents, and the ratio is the volume ratio of each solvent, such as "0-10% methanol / dichloromethane", which means that in the gradient elution process, the volume of methanol:dichloromethane in the mixed eluent is 0:100-10:100.

[0135] Preparation Example 1, synthesis of 2-isopropyl-4-methylpyridine-3-ol

[0136] A solution of 2-isopropyl-4-methylpyridine-3-amine (4 g, 26.63 mmol) in sulfuric acid (6.40 g, 65.25 mmol) and water (30 mL) was added dropwise with a solution of sodium nitrite (2.39 g, 34.62 mmol) in water (18 mL) at 0°C. The reaction was stirred at 0°C for 30 min, and then stirred at 25°C for 16 h. After the reaction was completed, the pH was adjusted to 7 with saturated aqueous sodium bicarbonate solution. The product was extracted with a mixed solvent of chloroform:isopropanol = 3:1 (100 mL*3), the organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent to obtain 2-isopropyl-4-methylpyridine-3-ol (4 g).

[0137] MS m / z (ESI): = 152.2 [M+H] + .

[0138] Example 1, 1-((12aS)-10-chloro-9-(2-fluoro-6-hydroxyphenyl)-7-((2-isopropyl-4- methylpyridin-3-yl)oxy)-3,4,12,12a-tetrahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepin- 2(1H)-yl)prop-2-en-1-one (Compound 1)

[0139]

[0140] Step 1: synthesis of methyl 2-fluoro-6-[(2-isopropyl-4-methylpyridin-3-yl)oxy]-3- nitrobenzoate (intermediate 1-2)

[0141] Dissolve the reactant 1-1 (6.7 g, 30.86 mmol) in DMF (30 ml), add sodium hydride (2.47 g, 61.72 mmol, effective content 60%) at 0 °C, and stir for 30 minutes at 0 °C. Dissolve 2-isopropyl-4-methylpyridin-3-ol (5.2 g, 34.39 mmol) in DMF (10 ml), add dropwise into the reaction bottle at 0 °C, and stir for 30 minutes at 0 °C. LCMS shows that the raw material reaction is complete. Pour the reaction liquid into saturated ammonium chloride solution (100 ml), extract with ethyl acetate (100 ml*2), wash with saturated brine, dry over anhydrous magnesium sulfate, and concentrate under reduced pressure. Purify the residue by flash silica gel column chromatography (eluent 10-20% ethyl acetate / petroleum ether gradient @ 60 mL / min). 80g Flash silica gel column, eluent 10-20% ethyl acetate / petroleum ether gradient @ 60 mL / min) to obtain the title compound (6 g).

[0142] MS m / z (ESI): = 349 [M+H] +

[0143] Step 2: Synthesis of methyl 3-amino-2-fluoro-6-[(2-isopropyl-4-methylpyridin-3- yl)oxy]benzoate (intermediate 1-3)

[0144] Dissolve intermediate 1-2 (2 g, 5.74 mmol) in methanol (30 ml), then add Pd / C (30.55 mg, 287.09 μmol), pass hydrogen gas (15 psi) through the hydrogen balloon, and stir for 16 hours at 25 °C. LCMS shows that the reaction is complete. Filter the reaction liquid through diatomite, then concentrate under reduced pressure. Purify the residue by flash silica gel column chromatography (eluent 30-40% ethyl acetate / petroleum ether gradient @ 18 ml / min). Obtain the title compound (800 mg). 20g Flash silica gel column, eluent 10-20% ethyl acetate / petroleum ether gradient @ 60 mL / min) to obtain the title compound (6 g).

[0145] MS m / z (ESI): = 349 [M+H] +

[0146] Step 3: Synthesis of methyl 3-amino-4-bromo-2-fluoro-6-[(2-isopropyl-4-methylpyridin- 3-yl)oxy]benzoate (intermediate 1-4)

[0147] Intermediate 1-3 (4.6 g, 14.45 mmol) was dissolved in acetic acid (40 mL) and then added portionwise with tribromopyridinium salt (6.01 g, 18.78 mmol) and stirred at 45 °C for 5 h. LCMS showed the reaction was completed. The reaction was poured into 20 wt% sodium sulfite solution and stirred for 10 min, extracted with ethyl acetate (10 mL*3), washed with saturated brine, and then the organic phase was concentrated under reduced pressure. The residue was purified by flash silica gel column (40 g, 30-40% ethyl acetate / petroleum ether gradient @ 36 ml / min). 40 g Flash silica gel column, eluent 30-40% ethyl acetate / petroleum ether gradient @ 36 ml / min). The title compound (4.8 g) was obtained.

[0148] MS m / z (ESI): = 399 [M+H] +

[0149] Step 4: Synthesis of methyl 4-bromo-3-chloro-2-fluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoate (Intermediate 1-5)

[0150] Copper chloride (3.25 g, 24.17 mmol) and tert-butyl nitrite (3.12 g, 30.21 mmol) were dissolved in acetonitrile (70 mL) and stirred for 10 min, and then intermediate 1-4 (4.8 g, 12.08 mmol) was dissolved in acetonitrile (50 mL) and added dropwise into the reaction flask at -20 °C, then warmed to 20 °C and stirred at 20 °C for 16 h. LCMS showed the starting material was consumed. The reaction was poured into 6 M aqueous hydrochloric acid solution (60 mL) and extracted with ethyl acetate (100 mL*3), dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by flash silica gel column (40 g, 10% methanol / dichloromethane gradient @ 100 mL / min). The title compound (4.9 g) was obtained. 40 g Flash silica gel column, eluent 30-40% ethyl acetate / petroleum ether gradient @ 36 ml / min). The title compound (4.8 g) was obtained.

[0151] MS m / z (ESI): = 417.8 [M+H] +

[0152] Step 5: Synthesis of 4-bromo-3-chloro-2-fluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoic acid (Intermediate 1-6)

[0153] Intermediate 1-5 (500 mg, 1.20 mmol) was dissolved in tetrahydrofuran (5 mL) and water (5 mL), after the addition of lithium hydroxide (287.40 mg, 12.00 mmol) it was stirred at 60 °C for 16 hours. LCMS showed the reaction was completed. The reaction solution was cooled to 25 °C and water (20 mL) was added, then ethyl acetate (20 mL) was added to extract. Subsequently, the aqueous phase was adjusted to pH 1-3 with 1M aqueous hydrochloric acid solution, extracted with isopropyl alcohol / dichloromethane 1:3, and the organic phase was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain the title compound (300 mg).

[0154] MS m / z (ESI): = 403.8 [M+H] +

[0155] Step 6: Synthesis of (S)-tert-butyl 4-[4-bromo-3-chloro-2-fluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoyl]-3-(hydroxymethyl)piperazine-1-carboxylate (Intermediate 1-7)

[0156] Intermediate 1-6 (300 mg, 745.08 μmol) was dissolved in dichloromethane (10 mL), and oxalyl chloride (283.71 mg, 2.24 mmol) was added. The mixture was stirred at 0 °C for 20 minutes, and LCMS showed that the starting material was completely converted. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in dichloromethane (10 mL). The dichloromethane solution of the residue was added dropwise to a dichloromethane solution containing (S)-tert-butyl 3-(hydroxymethyl)piperazine-1-carboxylate (193.37 mg, 894.09 μmol) and triethylamine (301.58 mg, 2.98 mmol) at 0 °C. After the dropwise addition was completed, the mixture was gradually warmed and stirred at 20 °C for 3 hours. LCMS showed that the reaction was completed. Water (15 mL) was added to quench the reaction, and dichloromethane (20 mL*3) was added to extract. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain the title compound (480 mg) as a crude product.

[0157] MS m / z (ESI): = 602 [M+H] +

[0158] Step 7: Synthesis of (S)-tert-butyl 9-bromo-10-chloro-7-((2-isopropyl-4-methylpyridin-3- yl)oxy)-6-oxo-3,4,12,12a-tetrahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepine-2(1H)- carboxylate (Intermediate 1-8)

[0159] Intermediate 1-7 (450 mg, 748.87 pmol) was dissolved in DMF (12 mL), sodium hydride (89.86 mg, 2.25 mmol, 60% of active content) was added at 0 °C, the reaction was stirred at 0 °C for 20 minutes. LCMS showed the starting material was consumed. The reaction was poured into an aqueous solution of ammonium chloride (30 mL), extracted with ethyl acetate (20 mL*3), washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by flash silica gel column (eluent 20-33% ethyl acetate / petroleum ether gradient @ 18 mL / min). 4 g flash silica gel column, eluent 20-33% ethyl acetate / petroleum ether gradient @ 18 mL / min). The title compound (220 mg) was obtained.

[0160] MS m / z (ESI): = 582 [M+H] +

[0161] Step 8: Synthesis of (S)-9-bromo-10-chloro-7-((2-isopropyl-4-methylpyridin-3- yl)oxy)-3,4,12,12a-tetrahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepine-2(1H)- carboxylic acid tert-butyl ester (Intermediate 1-9)

[0162] Intermediate 1-8 (220.00 mg, 378.72 pmol) was dissolved in tetrahydrofuran (15 mL), borane tetrahydrofuran (1 M, 7.57 mL) was added at 0 °C, then the reaction was heated to 60 °C and stirred for 16 hours. LCMS showed the reaction was completed. Methanol (15 mL) was slowly added to the reaction at 0 °C, then the temperature was raised to 60 °C, stirred for 1 hour, then cooled to 25 °C, and concentrated under reduced pressure. The residue was purified by thin layer preparative chromatography (silica, petroleum ether: ethyl acetate = 4:5). The title compound (180 mg) was obtained.

[0163] MS m / z (ESI): = 568 [M+H] +

[0164] Step 9: Synthesis of (12aS)-10-chloro-9-(2-fluoro-6-hydroxyphenyl)-7-((2- isopropyl-4-methylpyridin-3-yl)oxy)-3,4,12,12a-tetrahydro-6H-benzo[f]pyrazino[2,1- c][1,4]oxazepine-2(1H)-carboxylic acid tert-butyl ester (Intermediate 1-10)

[0165] Intermediate 1-9 (180 mg, 317.51 μmol) and 2-fluoro-6-hydroxyphenylpotassium trifluoroborate (346.08 mg, 1.59 mmol) were dissolved in toluene (6 mL), followed by the addition of potassium carbonate (131.64 mg, 952.53 μmol) and (2-dicyclohexylphosphino-2,6-dimethoxybiphenyl)[2-(2-amino-1,1- biphenyl)]palladium(II) methanesulfonate (SPhos Pd G3) (24.77 mg, 31.75 μmol), nitrogen, and then heating to 100 °C with stirring for 16 h. LCMS showed that the reaction was complete. The reaction solution was filtered with celite, extracted with ethyl acetate (10 mL), washed with water (10 mL), and concentrated to dryness under reduced pressure. The residue was purified twice by thin layer preparative chromatography (silica, ethyl acetate: petroleum ether = 2:1). The title compound (30 mg) was obtained.

[0166] MS m / z (ESI): = 598 [M+H] +

[0167] Step 10: Synthesis of 2-((S)-10-chloro-7-((2-isopropyl-4-methylpyridin-3-yl)oxy)- 1,2,3,4,12,12a-hexahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepin-9-yl)-3- fluorophenol (Intermediate 1-11)

[0168] Intermediate 1-10 was dissolved in dichloromethane (1 mL), followed by the addition of trifluoroacetic acid (770.00 mg, 6.75 mmol) and stirring at 25 °C for 2 h. TLC (petroleum ether: ethyl acetate 1:1) showed that the reaction was complete. The reaction solution was concentrated to dryness under reduced pressure to obtain the title compound (31 mg) as a crude product.

[0169] MS m / z (ESI): = 499 [M+H] +

[0170] Step 11: Synthesis of 1-((12aS)-10-chloro-9-(2-fluoro-6-hydroxyphenyl)-7-((2- isopropyl-4-methylpyridin-3-yl)oxy)-3,4,12,12a-tetrahydro-6H-benzo[f]pyrazino[2,1- c][1,4]oxazepin-2(1H)-yl)prop-2-en-1-one (Compound 1)

[0171] Intermediate 1-11 (31 mg, 50.65 pmol) was dissolved in dichloromethane (2 mL), diisopropylethylamine (19.64 mg, 151.96 pmol) was added to adjust pH = 8, and the solution was cooled to 0 °C. Acryloyl chloride (3.90 mg, 43.05 pmol) was dissolved in dichloromethane (0.2 mL) and added dropwise to the reaction bottle. The reaction solution was stirred at 16 °C for 16 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated to dryness under reduced pressure. The residue was purified by preparative high performance liquid chromatography (basic conditions, column: Boston Prime C18 150*30mm*5um; mobile phase: [A: water (0.04% ammonia water + 10 mM ammonium bicarbonate), B: acetonitrile]; B%: 43%-65%, 7 minutes) to obtain a pair of isomer (atropisomer) compound 1 isomer 1 (1.8 mg, LCMS retention time: 2.286 min), compound 1 isomer 2 (1.0 mg, LCMS retention time: 2.445 min).

[0172] LCMS detection conditions: Waters Xbridge C18 30*2.0mm, 3.5um; mobile phase: A) 0.04% trifluoroacetic acid in water; B) 0.02% trifluoroacetic acid in acetonitrile; gradient elution: mobile phase B increased from 0% to 95% in 5.8 minutes; mobile phase B maintained at 95% for 1.1 minutes; then mobile phase B decreased to 0% at 6.91 minutes and maintained at 0% for 0.09 minutes. The flow rate was 1.0 mL / min

[0173] MS m / z (ESI): = 552 [M+H] +

[0174] Compound 1 isomer 1 1H NMR: (400 MHz, Methanol-d4) δ 8.25 (d, J = 5.0 Hz, 1H), 7.22-7.17 (m, 1H), 7.16-7.08 (m, 1H), 6.89-6.71 (m, 1H), 6.61 (t, J = 7.4 Hz, 1H), 6.55 (t, J = 8.8 Hz, 1H), 6.24 (d, J = 16.1 Hz, 1H), 5.86 (d, J = 3.8 Hz, 1H), 5.78 (d, J = 10.0 Hz, 1H), 4.54-4.41 (m, 2H), 4.31-4.29 (m, 0.5H), 4.25-4.11 (m, 0.5H), 4.00 (d, J = 11.5 Hz, 1H), 3.93-3.78 (m, 2H), 3.60-3.47 (m, 0.5H), 3.38-3.33 (m, 0.5H), 3.28-3.12 (m, 1.5H), 3.06-3.00 (m, 1.5H), 2.95 (s, 1H), 2.66 (s, 1H), 2.21 (s, 1.5H), 2.15 (s, 1.5H), 1.30-1.19 (m, 3H), 1.16-1.09 (m, 3H).

[0175] Compound 1 Isomer 2 1 H NMR: (400 MHz, Methanol-d4) δ 8.25 (d, J = 5.0 Hz, 1H), 7.22-7.17 (m, 1H), 7.16-7.08 (m, 1H), 6.89-6.71 (m, 1H), 6.61 (t, J = 7.4 Hz, 1H), 6.55 (t, J = 8.8 Hz, 1H), 6.24 (d, J = 16.1 Hz, 1H), 5.86 (d, J = 3.8 Hz, 1H), 5.78 (d, J = 10.0 Hz, 1H), 4.54-4.41 (m, 2H), 4.31-4.29 (m, 0.5H), 4.25-4.11 (m, 0.5H), 4.00 (d, J = 11.5 Hz, 1H), 3.93-3.78 (m, 2H), 3.60-3.47 (m, 0.5H), 3.38-3.33 (m, 0.5H), 3.28-3.12 (m, 1.5H), 3.06-3.00 (m, 1.5H), 2.95 (s, 1H), 2.66 (s, 1H), 2.21 (s, 1.5H), 2.15 (s, 1.5H), 1.30-1.19 (m, 3H), 1.16-1.09 (m, 3H).

[0176] Example 2, 1-((12aS)-10-chloro-8-fluoro-9-(2-fluoro-6-hydroxyphenyl)-7-((2- isopropyl-4-methylpyridin-3-yl)oxy)-3,4,12,12a-tetrahydro-6H-benzo[f]pyrazino[2,1- c][l,4]oxazepin-2(lH)-yl)prop-2-en-l-one (Compound 2)

[0177]

[0178] Step 1: Synthesis of methyl 2,5-difluoro-6-((2-isopropyl-4-methylpyridin-3- yl)oxy)-3-nitrobenzoate (Intermediate 2-2)

[0179] To the reaction of 2-1 (3.60 g, 15.32 mmol) and 2-isopropyl-4-methylpyridin-3-ol (2.2 g, 14.55 mmol) in DMF (5 mL) was added sodium hydride (1.04 g, 26.04 mmol, 60% active content) at 0 °C with stirring. The reaction was stirred at 0 °C for 30 min. LCMS showed the starting material was consumed completely. Upon completion, the reaction was poured into saturated ammonium chloride solution (100 mL) and extracted with ethyl acetate (100 mL*2), washed with saturated brine, dried over anhydrous magnesium sulfate and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (silica gel, eluent 10-20% ethyl acetate / petroleum ether gradient @ 40 mL / min). 40 g Flash column chromatography (silica gel, eluent 10-20% ethyl acetate / petroleum ether gradient @ 40 mL / min). The title compound (4 g) was obtained.

[0180] MS m / z (ESI): = 367 [M+H] +

[0181] Step 2: Synthesis of methyl 3-amino-2,5-difluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoate (Intermediate 2-3)

[0182] Intermediate 2-2 (4 g, 10.92 mmol) was dissolved in methanol (100 mL) and wet palladium on carbon (1.16 g, 545.98 μmol, 5% active content) was added. The reaction was stirred at 20 °C under hydrogen atmosphere for 5 h. LCMS showed the starting material was consumed completely. The reaction was filtered through celite and the filtrate was concentrated under reduced pressure to give the title compound (3.5 g) as a crude product.

[0183] MS m / z (ESI): = 337 [M+H] +

[0184] Step 3: Synthesis of methyl 3-amino-4-bromo-2,5-difluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoate (Intermediate 2-4)

[0185] To a solution of intermediate 2-3 (3.5 g, 10.41 mmol) in acetic acid (50 mL) was added tribromopyridinium salt (3.66 g, 11.45 mmol) at 20 °C, warmed to 50 °C and stirred at 50 °C for 5 h, LCMS showed the reaction was complete. Concentrated to remove acetic acid under reduced pressure, quenched with sodium sulfite solution, adjusted pH to 8 with sodium carbonate solid, added ethyl acetate (100 mL) to the reaction, extracted with ethyl acetate (100 mL*2), dried over anhydrous magnesium sulfate, concentrated to dryness under reduced pressure. The residue was purified by flash silica gel column (eluent 20-30% ethyl acetate / petroleum ether gradient @ 40 mL / min) to give the title compound (4.2 g) as a crude product. 40g The residue was purified by flash silica gel column (eluent 20-30% ethyl acetate / petroleum ether gradient @ 40 mL / min) to give the title compound (4.2 g) as a crude product.

[0186] MS m / z (ESI): = 415.9 [M+H] +

[0187] Step 4: Synthesis of methyl 4-bromo-3-chloro-2,5-difluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoate (intermediate 2-5)

[0188] Copper chloride (2.66 g, 19.75 mmol) and tert-butyl nitrite (2.55 g, 24.69 mmol) were added to acetonitrile (40 mL), stirred at 20 °C for 10 min, intermediate 2-4 (4.1 g, 9.87 mmol) was dissolved in acetonitrile (40 mL) and added to the reaction flask at 0 °C, the reaction was stirred at 25 °C for 16 h. LCMS showed the starting material was consumed. The reaction was poured into 1M HC1 (20 mL), ethyl acetate (50 mL), water (30 mL), concentrated under reduced pressure, adjusted pH to 8 with sodium carbonate, extracted with ethyl acetate (50 mL*3), dried over anhydrous magnesium sulfate, concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (eluent 15% ethyl acetate / petroleum ether gradient @ 40 mL / min) to give the title compound (3.8 g). 40g The residue was purified by flash silica gel column (eluent 20-30% ethyl acetate / petroleum ether gradient @ 40 mL / min) to give the title compound (4.2 g) as a crude product.

[0189] MS m / z (ESI): = 435.8 [M+H] +

[0190] Step 5: Synthesis of 4-bromo-3-chloro-2,5-difluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoic acid (intermediate 2-6)

[0191] Intermediate 2-5 (3.8 g, 8.74 mmol) was dissolved in tetrahydrofuran (20 mL), water (20 mL), to which lithium hydroxide monohydrate (5.50 g, 131.14 mmol) was added, the reaction was stirred at 50 °C for 5 hours. LCMS showed the starting material was consumed, product was formed. The reaction was adjusted to pH 1 with 3M hydrochloric acid solution, filtered to get solid, the solid was dried to get 1.9 g, the filtrate was extracted with ethyl acetate (30 mL*3), dried over anhydrous magnesium sulfate, concentrated to dryness under reduced pressure to get solid (1.9 g), the two batches of solid were combined. The title compound (3.8 g, crude) was obtained.

[0192] MS m / z (ESI): = 421.8 [M+H] +

[0193] Step 6: Synthesis of (3S)-4-[4-bromo-3-chloro-2,5-difluoro-6-[(2-isopropyl-4- methylpyridin-3-yl)oxy]benzoyl]-3-(hydroxymethyl)piperazine-1 -carboxylic acid tert-butyl ester (Intermediate 2-8)

[0194] To a solution of Intermediate 2-6 (1.9 g, 4.52 mmol) in dichloromethane (20 mL) was added oxalyl chloride (1.72 g, 13.55 mmol) at 0 °C, the temperature was raised to 20 °C and stirred for 2 hours, LCMS showed the starting material was consumed, the solvent was removed by concentration under reduced pressure to get Intermediate 2-7; Intermediate 2-7 was dissolved in dichloromethane (15 mL), to the reaction was added dropwise a solution of (S)-3-(hydroxymethyl)piperazine-1 -carboxylic acid tert-butyl ester (1.07 g, 4.97 mmol) and triethylamine (1.83 g, 18.07 mmol) in dichloromethane (15 mL), the reaction was stirred for 3 hours. After the reaction was completed, water (20 mL) was added to the reaction, extracted with dichloromethane (20 mL*3), dried over anhydrous magnesium sulfate, concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (eluent: 15-40% ethyl acetate / petroleum ether gradient @ 40 mL / min). 40 g Flash silica gel column, eluent: 15-40% ethyl acetate / petroleum ether gradient @ 40 mL / min). The title compound (2.07 g) was obtained.

[0195] MS m / z (ESI): = 619.9 [M+H] +

[0196] Step 7: Synthesis of (S)-9-bromo-10-chloro-8-fluoro-7-((2-isopropyl-4-methylpyridin-3- yl)oxy)-6-oxo-3,4,12,12a-tetrahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepine-2(1H)- carboxylic acid tert-butyl ester (Intermediate 2-9)

[0197] To a solution of intermediate 2-8 (2 g, 3.23 mmol) in DMF (20 mL) was added sodium hydride (387.75 mg, 9.69 mmol, 60% active content) at 0 °C. The reaction was stirred at 0 °C for 2 h. LCMS showed the starting material was consumed. The reaction was poured into ammonium chloride solution (100 mL), extracted with ethyl acetate (100 mL*2), washed with saturated brine, and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (eluent: 15-35% tetrahydrofuran / petroleum ether gradient @ 35 mL / min). 20 g Flash column chromatography, eluent: 15-35% tetrahydrofuran / petroleum ether gradient @ 35 mL / min). The title compound (1.0 g) was obtained.

[0198] MS m / z (ESI): = 598 [M+H] +

[0199] Step 8: Synthesis of tert-butyl (12aS)-10-chloro-8-fluoro-9-(2-fluoro-6- hydroxyphenyl)-7-((2-isopropyl-4-methylpyridin-3-yl)oxy)-6-oxo-3,4,12,12a- tetrahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepine-2(1H)-carboxylate (Intermediate 2-10)

[0200] Intermediate 2-9 (250 mg, 417.44 µmol) and potassium 2-fluoro-6-hydroxyphenyl trifluoroborate (91.00 mg, 417.44 µmol) were dissolved in dioxane (6 mL) and water (1.5 mL), potassium carbonate (173.08 mg, 1.25 mmol), RuPhos (19.48 mg, 41.74 µmol) and methanesulfonic acid (2-dicyclohexylphosphino-2,6-diisopropoxy-1,1- biphenyl) (2-amino-1,1-biphenyl-2-yl)palladium(II) (RuPhos Pd G3) (34.91 mg, 41.74 µmol) were added, the reaction was stirred at 95 °C for 3 h under nitrogen atmosphere. The reaction was cooled to 20 °C, extracted with ethyl acetate (10 mL) for three times, the organic phase was combined and concentrated to dryness under reduced pressure (0.01 MPa). The residue was purified by column chromatography (eluent: 15-30% tetrahydrofuran / petroleum ether gradient @ 18 mL / min). 4 g Flash column chromatography, eluent: 15-30% tetrahydrofuran / petroleum ether gradient @ 18 mL / min). The title compound (160 mg) was obtained.

[0201] MS m / z (ESI): = 630.3 [M+H] +

[0202] Step 9: Synthesis of tert-butyl (12aS)-10-chloro-8-fluoro-9-(2-fluoro-6- hydroxyphenyl)-7-((2-isopropyl-4-methylpyridin-3-yl)oxy)-3,4,12,12a- tetrahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepine-2(1H)-carboxylate (Intermediate 2-11)

[0203] Intermediate 2-10 (20 mg, 31.74 μmol) was dissolved in tetrahydrofuran (3 mL), borane tetrahydrofuran solution (1 M, 634.84 uL) was added at 0 °C, the reaction was stirred at 40 °C for 16 h. Upon completion, it was cooled to 0 °C, methanol (5 mL) was added by syringe, and stirred at 25 °C for 1 h. The organic phase was concentrated under reduced pressure (0.01 MPa) to remove the solvent. The residue was purified by preparative thin layer chromatography (silica, tetrahydrofuran: petroleum ether = 1:1). The title compound was obtained as a crude product (60 mg).

[0204] MS m / z (ESI): = 616.1 [M+H] +

[0205] Step 10: Synthesis of 2-((S)-10-chloro-8-fluoro-7-((2-isopropyl-4- methylpyridin-3-yl)oxy)-1,2,3,4,12,12a-hexahydro-6H-benzo[f]pyrazino[2,1- c][1,4]oxazepin-9-yl)-3-fluorophenol (Intermediate 2-12)

[0206] Intermediate 2-11 (52.6 mg, 85.38 μmol) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (1.54 g, 13.51 mmol) was added, and the reaction was stirred at 15 °C for 1 h. The solvent was removed under reduced pressure (0.01 MPa). The title compound was obtained as a crude product (44.05 mg).

[0207] MS m / z (ESI): = 516 [M+H] +

[0208] Step 11: Synthesis of 1-((12aS)-10-chloro-8-fluoro-9-(2-fluoro-6- hydroxyphenyl)-7-((2-isopropyl-4-methylpyridin-3-yl)oxy)-3,4,12,12a- tetrahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepin-2(1H)-yl)prop-2-en-1-one (Compound 2)

[0209] Intermediate 2-12 (44 mg, 85.27 pmol) and triethylamine (69.03 mg, 682.20 pmol) were dissolved in dichloromethane (2 mL), acryloyl chloride (7.72 mg, 85.27 pmol) was added dropwise at 0 °C, the reaction was stirred at 0 °C for 1 h. The solvent was removed under reduced pressure (0.01 MPa). The residue was purified by preparative high-performance liquid chromatography (column: YMC-Actus Triart CI 8 100*30 mm*5 um; mobile phase: [A: water (0.05% ammonia water v / v), B: acetonitrile]; B%: 45%-65%, 10 min) to give a pair of isomers (atropisomers). Compound 2 Isomer 1 (1.5 mg, LCMS retention time: 1.306 min), Compound 2 Isomer 2 (1.5 mg, LCMS retention time: 2.615 min).

[0210] LCMS detection conditions: Waters Xbridge CI 8 30*2.0 mm, 3.5 um; mobile phase: A) 0.04% trifluoroformic acid in water; B) 0.02% trifluoroformic acid in acetonitrile; gradient elution: mobile phase B increased from 0% to 95% in 5.8 min; mobile phase B maintained at 95% for 1.1 min; followed by mobile phase B decreased to 0% in 6.91 min and maintained at 0% for 0.09 min. The flow rate was 1.0 mL / min

[0211] MS m / z (ESI): = 570.3 [M+H] +

[0212] Compound 2 Isomer 1 1 H NMR (400 MHz, METHANOL-d4) d 8.15 (dd, J = 1.8, 5.0 Hz, 1H), 7.26-7.15 (m, 1H), 7.09 (dd, J = 4.6, 8.4 Hz, 1H), 6.79 (d, J = 13.6 Hz, 1H), 6.68-6.57 (m, 2H), 6.24 (d, J = 17.3 Hz, 1H), 5.78 (s, 1H), 4.5-4.43 (m, 1H), 4.41-4.33 (m, 1H), 4.33-4.15 (m, 1H), 4.10-4.00 (m, 1H), 3.91-3.77 (m, 1H), 3.71 (m, 1H), 3.57-3.38 (m, 1H), 3.19-2.96 (m, 2H), 2.88 (m, 2H), 2.58 (m, 1H), 2.23-2.18 (s, 3H), 1.30-1.22 (m, 3H), 1.20-1.10 (m, 3H)

[0213] Compound 2 Isomer 2 1H NMR (400 MHz, METHANOL-d4) δ 8.15 (d, J = 5.0 Hz, 1H), 7.26-7.17 (m, 1H), 7.09 (d, J = 4.8 Hz, 1H), 6.80 (d, J = 8.5 Hz, 1H), 6.70-6.53 (m, 2H), 6.24 (d, J = 17.1 Hz, 1H), 5.78 (m, 1H), 4.45-4.35 (m, 2H), 4.33-4.15 (m, 1H), 4.05-3.90 (m, 1H), 3.89-3.78 (m, 1H), 3.75-3.60 (m, 1H), 3.56-3.41 (m, 1H), 3.35-3.13 (m, 1H), 3.12-2.99 (m, 1H), 2.92-2.80 (m, 2H), 2.65-2.59 (m 1H), 2.25-2.17 (m, 3H), 1.29-1.21 (m, 3H), 1.18-1.05 (m, 3H)

[0214] Example 3, (12aS)-2-acryloyl-10-chloro-8-fluoro-9-(2-fluoro-6-hydroxyphenyl)-7-((2- isopropyl-4-methylpyridin-3-yl)oxy)-1,2,3,4,12,12a-hexahydro-6H-benzo[f]pyrazino[2,1- c][1,4]oxazepin-6-one (Compound 3)

[0215]

[0216] Step 1: Synthesis of (12aS)-10-chloro-8-fluoro-9-(2-fluoro-6-hydroxyphenyl)-7-((2- isopropyl-4-methylpyridin-3-yl)oxy)-1,2,3,4,12,12a-hexahydro-6H-benzo[f]pyrazino[2,1- c][1,4]oxazepin-6-one (Intermediate 3-1)

[0217] To a solution of Intermediate 2-10 (80 mg, 126.97 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (14.48 mg, 126.97 μmol), and the reaction was stirred at 20 °C for 2 hours. LCMS showed the starting material was consumed completely. The reaction was concentrated to dryness under reduced pressure. The title compound was obtained as a crude product (81.7 mg).

[0218] MS m / z (ESI): = 530 [M+H] +

[0219] Step 2: Synthesis of (12aS)-2-acryloyl-10-chloro-8-fluoro-9-(2-fluoro-6- hydroxyphenyl)-7-((2-isopropyl-4-methylpyridin-3-yl)oxy)-1,2,3,4,12,12a- hexahydro-6H-benzo[f]pyrazino[2,1-c][1,4]oxazepin-6-one (Compound 3)

[0220] To a solution of intermediate 3-1 (81.7 mg, 126.87 pmol) in dichloromethane (3 mL) was added diisopropylethylamine (49.19 mg, 380.60 pmol) at 0 °C, adjusted pH to 8, a solution of acryloyl chloride 48 mg, 126.87 pmol) in dichloromethane was added to the reaction vial with stirring, the reaction was stirred at 0 °C for 20 min, then warmed to 20 °C, stirred for 1 h. LCMS showed the starting material was consumed. The reaction was concentrated to dryness under reduced pressure. The residue was purified by preparative high performance liquid chromatography (basic condition, column: Boston Prime C18 150*25mm*5um; mobile phase: [A: water (0.05% ammonia water v / v), B: acetonitrile]; B%: 35%-57%, 7 min), then further purified by supercritical fluid chromatography (condition: column: DAICEL CHIRALCEL OJ (250mm*30mm, 10um); mobile phase: [B: ethanol solution containing 0.1% ammonia water]; B%: 25%-25%), to give a pair of isomers (atropisomers). Compound 3 Isomer 1 (5.4 mg, LCMS retention time: 2.420 min), Compound 3 Isomer 2 (6.2 mg, LCMS retention time: 2.699 min).

[0221] LCMS detection condition: Waters Xbridge C18 30*2.0mm, 3.5um; mobile phase: A: 0.04% trifluoroacetic acid in water; B: 0.02% trifluoroacetic acid in acetonitrile; gradient elution: mobile phase B increased from 0% to 95% in 5.8 min; mobile phase B kept at 95% for 1.1 min; followed by mobile phase B decreased to 0% in 6.91 min and maintained for 0.09 min. The flow rate was 1.0 mL / min

[0222] MS m / z (ESI): = 585 [M+H] +

[0223] Compound 3 Isomer 1 1H NMR (400 MHz, Methanol-d4) δ 8.15 (s, 1H), 7.25 (d, J = 7.9 Hz, 1H), 7.07 (s, 1H), 6.85 - 6.58 (m, 3H), 6.32 (d, J = 16.6 Hz, 1H), 5.84 (d, J = 10.5 Hz, 1H), 4.25 (s, 3H), 4.05 (s, 2H), 3.99 - 3.83 (m, 1H), 3.83 - 3.71 (m, 1H), 3.69 - 3.45 (m, 2H), 2.34 (s, 1.5H), 2.16 - 2.03 (m, 1.5H), 1.37 - 1.26 (m, 3H), 1.20 - 1.13 (m, 3H)

[0224] Compound 3 Isomer 2 1 H NMR (400 MHz, Methanol-d4) δ 8.16 (d, J = 4.9 Hz, 1H), 7.26 (s, 1H), 7.11 (s, 1H), 6.86 - 6.59 (m, 3H), 6.32 (d, J = 16.8 Hz, 1H), 5.84 (d, J = 11.4 Hz, 1H), 4.26 (s, 3H), 4.15 - 4.02 (m, 2H), 4.00 - 3.83 (m, 1H), 3.81 - 3.61 (m, 2H), 3.61 - 3.45 (m, 1H), 2.66 (s, 1H), 2.34 (s, 2H), 2.16 - 2.00 (m, 2H), 1.42 - 1.28 (m, 3H), 1.19 - 1.12 (m, 3H)

[0225] Test Example, Inhibition Test of Human Non-Small Cell Lung Cancer H358 Cell Proliferation

[0226] Brief Description of Test Principle:

[0227] KRAS mutation is a driving factor for abnormal proliferation of various tumor cells, among which KRAS-G12C mutation occurs at a high rate in non-small cell lung cancer and pancreatic cancer. In vitro, human non-small cell lung cancer H358 carrying KRAS-G12C mutation was used as a cell model to detect the inhibitory activity of KRAS-G12C inhibitor on cell proliferation. The experimental method is described in the reference: Janes MR et al, Targeting KRAS Mutant Cancers with a Covalent G12C-Specific Inhibitor, Cell 2018 Jan 25; 172(3): 578-589.

[0228] Experimental Materials and Instruments:

[0229] NCI-H358 cells were purchased from ATCC (USA);

[0230] 384-well plates were purchased from Corning (USA);

[0231] RPMI-1640 medium was purchased from Gibco (USA);

[0232] DMEM medium was purchased from Gibco (USA);

[0233] FBS was purchased from Gibco (USA);

[0234] Horse serum was purchased from Gibco (USA);

[0235] Penicillin-streptomycin was purchased from Invitrogen (USA);

[0236] 1640 complete medium: RPMI1640 medium + 10% FBS + 1% Penicillin-streptomycin;

[0237] DMEM complete medium: DMEM + 10% FBS + 2.5% horse serum + 1% Penicillin-streptomycin;

[0238] Celltiter Glo assay kit (2D-CTG reagent) was purchased from Promega (USA);

[0239] Echo 550 Liquid Handler (Labcyte, USA);

[0240] Plate reading instrument: Envision (PerkinElmer, USA).

[0241] Experimental method:

[0242] H358 cells were collected on day 1, counted, adjusted cell density, and cell suspension was seeded at 40 μL into 384-well plates (containing 800 cells / well) and incubated at 37 °C, 5% CO2 incubator overnight. On day 2, compound dilution and cell treatment. Compound stock 10 mM was diluted with DMSO to 200* final concentration (e.g. compound final concentration is 1 μM, first dilute 10 mM to 200 μM, take 2 μL to 98 μL DMSO, mix well, then take 40 μL 200 μM to source plate), using ECHO 550 gradient dilution program, compound was added into the cell culture plate seeded on day 1 with gradient concentration (200 nL / well, each concentration of test compound). The cell culture plate was placed in a 37 °C, 5% CO2 incubator for continued incubation for 3 days. On day 5, the cell culture plate was removed and added 2D-CTG reagent 20 μL / well, incubated at room temperature for 20 min. Placed in Envision to read Luminescence signal.

[0243] Data analysis:

[0244] Cell proliferation inhibition rate % = (Average_DMSO - Sample) / Average_DMSO - Ratio_PositiveControl) x 100%. 100% inhibition is defined as the level of cell proliferation inhibition under the treatment of 1 μM positive compound (2-((S)-1-acryloyl-4-(7-(3-hydroxynaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile). Data analysis was performed using XLfit. Concentration-effect curve was fitted using non-linear four-parameter curve, and IC 50 :

[0245] Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC 50 -X) * HillSlope))

[0246] X: Log of compound concentration

[0247] Y: Percent inhibition (% inh)

[0248] Test results:

[0249] Under the conditions of this test, the tested compounds have significant inhibitory activity on the proliferation of human non-small cell lung cancer H358 cells with KRAS G12C mutation. The binding activity of the tested compounds to KRAS G12C protein is shown in Table 1.

[0250] Table 1

[0251] Compound IC 50 ]]> Compound 1 Isomer 1 271.7 nM Compound 2 Isomer 1 396.3 nM Compound 3 Isomer 1 803.3 nM

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof: in, R 1 Selected from R 1a Substituted with the following groups: C6-C 10 aryl; Every R 1a is selected from F, Cl, Br, I, OH, CN, NH2, C1-C6 alkyl, C1-C6 alkoxy, NH(C1-C6 alkyl); R 2 、R 3 independently selected from H or halogen; R 4a 、R 4b Selected from H, or R 4a 、R 4b Together form = O; R 5 Selected from R 10 Selected from C1-C3 alkyl; n is selected from 0; L is selected from O; R 11 Selected from R 11a Substituted: 5-6 membered heteroaryl; Every R 11a Selected from F, Cl, Br, I, OH, CN, NH2, C1-C6 alkyl, C1-C6 alkoxy, NH(C1-C6 alkyl).

2. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 Selected from R 1a Substituted with the following groups: phenyl.

3. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1a Selected from F or OH.

4. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 Selected from 5. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 2 、R 3 Independently selected from H, F, Cl.

6. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 2 is selected from H or halogen.

7. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 3 Selected from halogen.

8. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 11 Selected from R 11a Substituted with the following groups: pyridyl.

9. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 11a Selected from F, Cl, Br, I, OH, CN, NH2, C1-C6 alkyl, C1-C6 alkoxy.

10. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 9, characterized in that: R 11a Selected from C1-C3 alkyl.

11. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 10, characterized in that: R 11a is selected from methyl or isopropyl.

12. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 11 Selected from 13. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

14. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

15. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

16. A pharmaceutical composition comprising the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

17. Use of the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 16, in the preparation of a medicament for preventing or treating a KRAS G12C-related disease.

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