RAS inhibitors

Abstract

The present invention provides compounds having the structure of formula (I) useful as RAS inhibitors, pharmaceutical compositions comprising such compounds, methods of preparing such compounds, and uses of such compounds in the treatment of cancer. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry. More specifically, the present invention relates to a class of compounds with novel structures that can be used as RAS inhibitors, pharmaceutical compositions containing such compounds, methods for preparing such compounds, and the use of these compounds in the treatment of cancer or tumors. Background Art

[0002] RAS, namely rat sarcoma oncogene homolog, represents a group of closely related monomeric globular proteins belonging to the GTPase protein family. Specifically, under normal physiological conditions, RAS is activated by receiving growth factors and various other extracellular signals and is responsible for regulating functions such as cell growth, survival, migration, and differentiation. These regulatory functions of RAS are carried out through the conversion between the GDP-bound state and the GTP-bound state, namely the "molecular switch" (Alamgeer et al., Current Opin Pharmacol. 2013, 13: 394-401). RAS in the GDP-bound form is in an inactive state, dormant or off, at which time the signal system is off and it will be activated when exposed to some growth-promoting stimuli. For example, it can be induced by a guanine nucleotide exchange factor (GEF) to release GDP and bind to GTP. As a result, RAS is "turned on" thereby and transformed into the active form of RAS, which recruits and activates various downstream effectors to conduct signal transduction, capable of transmitting signals on the cell surface into the cytoplasm, thereby controlling numerous key cell processes such as differentiation, survival, and proliferation (Zhi Tan et al., Mini-Reviews in Medicinal Chemistry, 2016, 16, 345-357).

[0003] RAS has GTPase activity and it can cleave the terminal phosphate of GTP to convert it into GDP, that is, convert itself into an inactive state. However, the endogenous GTPase activity of RAS is very low, and the conversion of GTP-RAS to GDP-RAS requires an exogenous protein GAP (GTPase-activating protein). GAP interacts with RAS and promotes the conversion of GTP to GDP. Therefore, any RAS gene mutation that affects the interaction between RAS and GAP or affects the conversion of GTP to GDP will cause RAS to be in an activated state for a long time, thereby continuously transmitting signals of growth and division to the cells, stimulating the cells to proliferate continuously, and ultimately leading to the formation and development of tumors.

[0004] Among the genes associated with human tumors, there are three ubiquitously expressed RAS genes, H-RAS, K-RAS, and N-RAS, which encode highly homologous HRAS, NRAS, and KRAS proteins of approximately 21 KDa, respectively. In 1982, researchers first discovered the mutational activation of RAS in cancer cell lines (Chang, E.H. et al., Proceedings of the National Academy of Sciences of the United States of America, 1982, 79(16), 4848-4852). Subsequently, large-scale genomic sequencing studies conducted in different cancer types have revealed that RAS proteins are mutated in more than 30% of cancer types, especially with the highest mutation rates in pancreatic cancer (>90%), colorectal cancer (45%), and lung cancer (35%). Transgenic and genetically engineered mouse models have also shown that mutant RAS proteins are sufficient to drive and initiate various types of cancers, and the RAS oncogene is also crucial for the maintenance and progression of tumors in multiple cancer types. For example, in RAS mutant cancer cell lines and cancer animal models, it has been shown that RNA interference can slow down tumor growth. These studies have made RAS tumor proteins an attractive and widely accepted anti-cancer drug target in the pharmaceutical field.

[0005] Studies have shown that RAS mutations are most common in KRAS, and KRAS mutations can be observed in approximately 85% of cancers driven by RAS mutations; the vast majority of RAS mutations occur at codons G12, G13, and Q61, and approximately 80% of KRAS mutations occur at the glycine at codon 12, such as G12C mutations, G12D mutations, G12V mutations, G12A mutations, G12R mutations, G12S mutations, G13D mutations, and Q61H mutations. KRAS mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and cholangiocarcinoma, and are also found in 25% of patients with non-small cell lung cancer (McCormick, F. et al., Clinical Cancer Research 21(8), 1797-1801, 2015). Therefore, mutant KRAS proteins have become the most important branch in the research of Ras drug targets, and the development of its inhibitors is also regarded as a very promising research direction in anti-cancer / tumor drug development.

[0006] However, drug development targeting RAS in the past few decades has shown that due to the smooth surface of the RAS protein, the lack of obvious groove-like or pocket-like structures for binding small molecule inhibitors, and its very high affinity for guanine substrates (picomolar level), the development of small molecule inhibitors of RAS has fallen into an insoluble dilemma. Therefore, RAS has long been considered an "undruggable" target in the industry. At the same time, there is still a great need for compounds of more structural types or modes as KRAS inhibitors to provide more treatment options, or to provide further improved inhibitory activity compared to existing KRAS inhibitors, so as to provide more potent therapeutic drugs for clinical use.

[0007] The present invention solves these and other needs. The present invention provides novel structure inhibitor compounds having RAS protein inhibitory activity. Due to their improved structural modes, these compounds of the present invention have enhanced activity in inhibiting the RAS protein and related tumor inhibitory activity compared to the existing RAS protein inhibitors in the prior art, have good pharmacokinetic properties, and thus have good druggability, such as being more easily absorbed in the body after administration in a convenient manner, with reduced toxic and side effects, improved drug resistance and safety, and reduced risk of drug interactions. Brief Description of the Invention

[0008] The present invention provides compounds having the structural formula (I) as defined below, their stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates:

[0009]

[0010] Wherein the definitions of each group are as defined in the detailed description of the invention.

[0011] The present invention also provides a pharmaceutical composition comprising a compound of the present invention or its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates and optionally a pharmaceutically acceptable excipient or carrier.

[0012] The present invention also provides a compound of the present invention or its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates for use as a drug.

[0013] The present invention also provides a compound of the present invention or its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates for use as an inhibitor of RAS protein, especially KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS amplified cells.

[0014] The present invention also provides the compounds of the present invention or their stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions containing the same for the treatment and / or prevention of diseases mediated by RAS proteins, especially KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS amplification.

[0015] The present invention also provides the use of the compounds of the present invention or their stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions containing the same for the treatment and / or prevention of diseases mediated by RAS proteins, especially KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS amplification.

[0016] The present invention also provides the use of the compounds of the present invention or their stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions containing the same in the preparation of a medicament for the treatment and / or prevention of diseases mediated by RAS proteins, especially KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS amplification.

[0017] The present invention also provides a method for the treatment and / or prevention of diseases mediated by RAS proteins, especially KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS amplification, comprising administering a therapeutically effective amount of the compounds of the present invention or their stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions containing the same to a subject in need thereof.

[0018] The present invention also provides a method for treating a tumor or cancer, which comprises administering the compounds of the present invention or their stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions containing the same to a patient in need thereof.

[0019] The present invention also provides the use of the compounds of the present invention or their pharmaceutically acceptable salts or solvates as research tool compounds for RAS inhibitors, particularly for inhibiting KRAS mutant proteins (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutant proteins) and KRAS amplification in research.

[0020] The present invention also provides a pharmaceutical combination, which comprises the compounds of the present invention, their stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates and one or more other pharmaceutically active agents.

[0021] The present invention also provides a method for preparing the compounds of the present invention. Detailed Description of the Invention

[0022] Definition

[0023] Unless otherwise indicated, each term used in the specification and claims has the meaning shown below. In the case where a specific term or phrase is not specifically defined, it should be understood in accordance with the ordinary meaning in the art. In case of conflict, the present specification (including the definitions) shall prevail.

[0024] In the case of conflict between the chemical structure and the name of the compounds disclosed herein, the chemical structure shall prevail.

[0025] As used herein, the term "RAS mutation" or "RAS mutant protein" refers to a protein encoded and expressed by a RAS gene in which one or more codons are mutated, typically including but not limited to RAS proteins in which glycine at codon 12, glycine at codon 13 or glutamine at codon 61 of RAS are mutated, such as mutant HRAS, NRAS or KRAS. These residues are located at the active site of RAS, and mutations thereof can impair the intrinsic or GAP-catalyzed GTPase activity of RAS, resulting in the persistent presence of RAS bound to GTP.

[0026] For the purposes of the present invention, the terms "RAS mutation" or "RAS mutant protein" and "RAS" when describing inhibitory activity are used interchangeably and generally refer to mutant HRAS, NRAS or KRAS, such as but not limited to KRAS-G12C (mutation of glycine to cysteine at codon G12), KRAS-G12D (mutation of glycine to aspartic acid at codon G12), HRAS-G12D, NRAS-G12D, KRAS-G12V (mutation of glycine to valine at codon G12), KRAS-G13D (mutation of glycine to aspartic acid at codon G13); particularly refers to KRAS mutant proteins, more particularly to KRAS-G12C mutant proteins, KRAS-G12D mutant proteins, KRAS-G12V mutant proteins, G12A mutant proteins, G12R mutant proteins, G12S mutant proteins, KRAS-G13D mutant proteins and Q61H mutant proteins.

[0027] As used herein, the term "treatment" means administering to a subject, such as a mammal, such as a human, having the disease or symptoms of the disease, one or more of the compounds of the present invention or pharmaceutically acceptable salts or solvates thereof described herein, for the purpose of curing, alleviating, reducing or affecting the disease or the symptoms of the disease. Preferably, the treatment is curative or ameliorative.

[0028] As used herein, the term "prevention" is well known in the art and means administering to a subject, such as a mammal, such as a human, suspected of having or susceptible to a Ras-mediated disease, particularly cancer or tumor, as defined herein, one or more of the compounds or pharmaceutically acceptable salts or solvates thereof described herein, such that the risk of developing the defined disease is reduced or the onset of the disease is prevented. The term "prevention" encompasses the use of the compounds of the present invention prior to the diagnosis or determination of any clinical and / or pathological symptoms.

[0029] As used herein, the terms "inhibit" and "reduce" or any variants of these terms refer to the ability of a bioactive agent to reduce the signal transduction activity of a target by directly or indirectly interacting with the target, and refer to any measurable reduction or complete inhibition of the activity of the target. For example, compared to normal conditions, it can be a reduction in activity (e.g., KRAS activity) of about, at most about or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more, or any range derivable therefrom.

[0030] As used herein, the term "RAS-mediated disease" refers to a disease in which RAS promotes the occurrence and development of the disease, or in which inhibition of RAS will reduce the incidence of the disease and reduce or eliminate the symptoms of the disease. For the purposes of the present invention, "RAS-mediated disease" preferably refers to a KRAS-mediated disease, and more preferably a KRAS mutation-mediated cancer or tumor.

[0031] As used herein, the term "cancer" or "tumor" refers to abnormal cell growth and proliferation, whether malignant or benign, and all pre-cancerous cells and cancer cells and tissues. For all aspects of the present invention, the cancer or tumor includes, but is not limited to, lung adenocarcinoma, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, melanoma of the skin or eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal cell cancer, renal pelvic cancer, central nervous system tumor (CNS), primary CNS lymphoma, spinal tumor, brainstem glioma or pituitary adenoma.

[0032] For all aspects of the present invention, preferably, the cancer or tumor is related to RAS, especially KRAS mutation and amplification, including but not limited to the above-mentioned tumor types and their preferred ranges. Particularly preferred tumors of the present invention include lung cancer, lung adenocarcinoma, colon cancer, rectal cancer, pancreatic cancer, endometrial cancer, cholangiocarcinoma, leukemia and ovarian cancer.

[0033] As used herein, the terms "subject", "individual" or "patient" refer to a vertebrate. In certain embodiments, the vertebrate is a mammal. Mammals include, but are not limited to, farm animals (such as cows), sports animals, pets (such as guinea pigs, cats, dogs, rabbits and horses), primates, mice and rats. In certain embodiments, the mammal is a human.

[0034] As used herein, the term "therapeutically effective amount" refers to an amount or dose that is generally sufficient to produce a beneficial therapeutic effect on a patient with a "RAS-mediated disease" such as cancer or tumor in need of treatment. Those skilled in the art can determine the effective amount or dose of the active ingredient in the present invention by conventional methods in combination with conventional influencing factors.

[0035] As used herein, the term "drug combination" means that the compounds of the present invention can be combined with other active agents for the purposes of the present invention. The other active agents can be one or more additional compounds of the present invention, or can be a second or additional (e.g., third) compound that is compatible with the compounds of the present invention, i.e., does not adversely affect each other, or has complementary activities. For example, these active agents are known to regulate other biological activity pathways, or regulate different components in the biological activity pathways involved by the compounds of the present invention, or even overlap with the biological targets of the compounds of the present invention. Such active agents are suitably combined in an effective amount to achieve the desired purpose. The other active agents can be co-administered with the compounds of the present invention in a single pharmaceutical composition, or administered separately from the compounds of the present invention in different discrete units, and when administered separately, can be administered simultaneously or sequentially. The sequential administration can be close or distant in time.

[0036] As used herein, the term "pharmaceutically acceptable" means molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered in an appropriate amount to an animal, such as a human.

[0037] As used herein, the term "pharmaceutically acceptable salt" refers to those salts that retain the biological effectiveness and properties of the parent compound and are not undesirable in a biological or other respect, including acid addition salts and base addition salts. "Pharmaceutically acceptable acid addition salts" can be formed from compounds having basic groups and inorganic acids or organic acids. Inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, etc., and organic acids can be selected from aliphatic, alicyclic, aromatic, arylaliphatic, heterocyclic, carboxylic, and sulfonic organic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc. "Pharmaceutically acceptable base addition salts" include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, etc., and salts derived from pharmaceutically acceptable organic non-toxic bases, including but not limited to primary, secondary, and tertiary amines, substituted ammonium, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, tromethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, triethanolamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc.

[0038] As used herein, the term "isomer" refers to any possible stereoisomers, enantiomeric mixtures, including racemates, diastereomeric mixtures, geometric isomers, atropisomers, and / or tautomers of a compound. Methods for determining and separating the stereochemistry of the isomers are well known to those skilled in the art (S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994).

[0039] Certain compounds of the present invention contain at least one asymmetric center and thus give rise to stereoisomers. Accordingly, the present invention encompasses all possible isomeric forms of the compounds defined herein, as well as their pharmaceutically acceptable salts or solvates, unless otherwise indicated.

[0040] As used in the structural formulas or structural fragments of the compounds herein or represents the absolute configuration of a stereocenter, i.e., a chiral center, and accordingly, in the naming of the compounds or intermediates provided by the present invention, the absolute configuration of the chiral center is represented by R or S; in some definitions of the compounds of the present invention, atropisomerism can also be used to represent the configuration of the compound, and the determination of these configurations uses the Cahn-Ingold-Prelog rules well known to those skilled in the art.

[0041] It should be understood that when those skilled in the art can judge that the compound has and only has a pair of chiral isomers based on the compound structure shown herein, and it can be easily resolved based on the conventional methods in the art, then the disclosure of the racemate of the compound herein (whether in the structural formula or chemical name) should be regarded as having separately disclosed each isomer of the compound.

[0042] As used in the structural fragments involved herein indicates that the bond crossing therewith is the bond by which the structural fragment is attached to the rest of the molecule.

[0043] Unless otherwise defined, substituents shown across chemical bonds in the cyclic structural fragments involved herein, such as -(R3) in m means that the defined number of substituents can be substituted at one or more sites available in the ring, where m = 0 means that the ring does not carry non-H substituents, but the ring atoms still carry hydrogen atoms in the stoichiometry of the chemical valence.

[0044] The compounds of the present invention include the unlabeled forms of the compounds of the present invention and their isotopically labeled forms. Isotopically labeled forms of the compounds are different compounds in which only one or more atoms are replaced with the corresponding isotopically enriched atoms. Examples of isotopes that can be incorporated into the compounds of the present invention include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 O, 17 O, 35 S, 18 F, 37 Cl and 125 I. Such isotopically labeled compounds can be used, for example, as probes in biological assays, analytical tools, or as therapeutic agents. In certain embodiments, the compounds of the present invention are provided in unlabeled form.

[0045] In some embodiments, the compounds of the present invention are provided in unlabeled form. In other embodiments, the compounds of the present invention are provided in isotopically labeled form, such as compounds in which one or more H atoms are replaced with deuterium atoms (D), such as in the moiety in which each of the two R2s can independently be D, or can be each of the defined groups substituted with D; R4 can be each of the defined groups substituted with D; for example, each of the groups defined as R7 can independently be optionally substituted with one or more isotopes, such as with D.

[0046] As used herein, the term "solvate" refers to a solvate addition form of a compound that contains a stoichiometric or non-stoichiometric amount of a solvent, including any solvated form of the compounds of the present invention, including, for example, solvates with water, such as hydrates, or solvates with organic solvents, such as methanol, ethanol, or acetonitrile, i.e., as methanolates, ethanolates, or acetonitriles, respectively; or in the form of any polymorph. It should be understood that such solvates of the compounds of the present invention also include solvates of the pharmaceutically acceptable salts of the compounds of the present invention.

[0047] As used herein, the term "metabolite" means a product generated by the in vivo metabolism of a compound. Such products can, for example, result from the oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, etc. of the administered compound. Identification and analysis of metabolite products are carried out in a manner well known to those skilled in the art.

[0048] As used herein, the term "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and have sufficient purity and sufficiently low toxicity. Examples thereof include, but are not limited to, cellulose and its derivatives (such as sodium carboxymethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as magnesium stearate), calcium sulfate, vegetable oils, polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tweens), wetting agents (such as sodium dodecyl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, etc.

[0049] As used herein, the term "halogen" or "halo" means F, Cl, Br or I. In addition, the term "halogen-substituted" group used when defining a group herein is intended to include mono-halogenated or poly-halogenated groups, in which one or more of the same or different halogens replace one or more hydrogens in the corresponding group.

[0050] As used herein, the term "alkyl" means a straight-chain or branched-chain monovalent saturated hydrocarbon group composed of carbon atoms and hydrogen atoms. Generally, an alkyl group has 1 to 10, for example 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3 or 1 to 2 carbon atoms. For example, as used herein, the term "C 1-6 alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms, and further preferably "C 1-3 alkyl"; in some embodiments, for the convenience of definition, the term "C 0-6 alkyl" means that the alkyl group is absent or is a straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms; examples thereof include methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, sec-butyl or tert-butyl), pentyl (including n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylpentyl, etc.

[0051] In the definition of the compounds herein, "alkyl" as a separate substituent, such as -C 1-6 alkyl or "alkyl" as part of a substituent, such as -OC 1-6 alkyl in -OC 1-6 alkyl, is optionally substituted, and when present, one or more (for example 1, 2, 3, 4 or 5) hydrogen atoms are optionally replaced by the defined substituents. When there are more than one substituent, they may be the same or different and may be located on the same or different C atoms. The substituents are selected from one or more of the following: D, OH, NH2, halogen, CN, -O-C 1-6 alkyl, -O-CON(H or -C 1-6The alkyl group is further optionally substituted by halogen or D. Examples of the optionally substituted alkyl group include, but are not limited to, -CH2Cl, -CH2F, -CHF2, -CF3, -CCl3, -C2F5, -C2Cl5, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C(CH3)2CF3, -CF(CF3)2, -CH2CN, -CH2CH2CN, -CH2D, -CHD2, -CD3, -CH2CD3, -CH2 - OCH3, -CH2CH2 - OCH3, -CH2 - OCF3, -CH2 - OCD3, -CH2 - OCH2CH3, -CH2CH2 - O - CH3, -CH2CH2 - O - CH2CH3, -CH2 - OCONH2, -CH2 - OCONH(CH3), -CH2 - OCON(CH3)2, -CH2 - OCON(CH2CH3)(CH3), -CH2 - OCON(CH2CH3)(CF3).

[0052] As used herein, the term "alkylene" means a straight-chain or branched-chain divalent saturated hydrocarbon group composed of carbon atoms and hydrogen atoms, preferably a straight-chain divalent saturated hydrocarbon group. Generally, the alkylene has 1 to 10, for example 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 carbon atoms. For example, as used herein, the term "-C 1-3 alkylene-" or "-(CH2) n -, where n is an integer from 0 to 3" means that the alkylene does not exist or is a straight-chain or branched-chain divalent saturated hydrocarbon group having 1 to 3 carbon atoms; the term "-C 1-6 alkylene-" means a straight-chain or branched-chain divalent saturated hydrocarbon group having 1 to 6 carbon atoms. Unless otherwise defined, the "alkylene" in the definitions of the compounds herein is optionally substituted, for example, by one or more D, OH, NH2, halogen, CN, or -C 1-6 alkyl optionally substituted by halogen or -O - C 1-6 alkyl substituted.

[0053] As used herein, the term "alkoxy" means an alkyl group as defined herein that is connected to the rest of the molecule through an oxygen atom. Specifically, the alkoxy has 1 - 10, for example 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 carbon atoms. For example, as used herein, the term "C 1-6 alkoxy" or "-O - C 1-6 alkyl" means a straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms that is connected to the rest of the molecule through an oxygen atom, and further preferably "-C 1-3 alkoxy" or "-O - C 1-3"Alkyl", examples thereof include -O-methyl, -O-ethyl, -O-propyl (including -O-n-propyl and -O-isopropyl), -O-butyl (including -O-n-butyl, -O-isobutyl, -O-sec-butyl or -O-tert-butyl), -O-pentyl (including -O-n-pentyl, -O-isopentyl, -O-neopentyl), -O-n-hexyl, 2-methylpentyl -O-, etc. Unless otherwise defined, the "alkoxy group" as a substituent in the definition of the compounds in this document is optionally substituted, that is, the alkyl part therein is optionally substituted, for example, by one or more D, OH, NH2, halogen, CN or -O-C optionally substituted by halogen or D 1-6 substituted by alkyl, examples include but are not limited to -OCH3, -OCH2CH3, -OCD3, -OCH2CD3, -OCH2CH2CN, -OCH2CH2CH3, -OCH(CH3)(CH3), -OCH2Cl, -OCH2F, -OCHF2, -OCF3, -OCH2CH2F, -OCH2CHF2, -OCH2CF3, -OCH2CH2CH2F, -OCH2CH2CHF2, -OCH2CH2CF3, -O-CH2-OCH3, -OCH2CH2-OCH3, -OCH2-OCF3, -OCH2-OCD3, -OCH2-OCH2CH3, -OCH2CH2-O-CH2CH3.

[0054] As used herein, the term "alkenyl" refers to a straight-chain or branched-chain unsaturated hydrocarbon group composed of carbon atoms and hydrogen atoms and containing at least one double bond. Specifically, alkenyl has 2-8, for example 2 to 6, 2 to 5, 2 to 4 or 2 to 3 carbon atoms. For example, as used herein, the term "C 2-6 alkenyl" refers to a straight-chain or branched-chain alkenyl having 2 to 6 carbon atoms, and further preferably "C 2-4 alkenyl", such as vinyl, propenyl, allyl, butenyl, pentenyl, etc. The carbon atom in the alkenyl group connected to the rest of the molecule can be saturated or an olefinic carbon atom.

[0055] As used herein, the term "alkynyl" refers to a straight-chain or branched-chain unsaturated hydrocarbon group composed of carbon atoms and hydrogen atoms and containing at least one triple bond. Specifically, alkynyl has 2-8, for example 2 to 6, 2 to 5, 2 to 4 or 2 to 3 carbon atoms. For example, as used herein, the term "C 2-6 alkynyl" refers to a straight-chain or branched-chain alkynyl having 2 to 6 carbon atoms, and further preferably "C 2-4 alkynyl", such as ethynyl, propynyl, propargyl, butynyl, etc. The carbon atom in the alkynyl group connected to the rest of the molecule can be saturated or an alkynyl carbon atom.

[0056] Unless otherwise defined, the "alkenyl" and "alkynyl" groups as substituents in the definitions of the compounds herein are optionally substituted, and the substituents can be selected from one or more of the following: D, halogen, CN, OH, -O-C 1-6 alkyl, -O-CON(H or -C 1-6 alkyl)2, wherein the alkyl is further optionally substituted with halogen or D. Examples include, but are not limited to, -CH=CH2, -CH=CHF, -CH=CF2, -CF=CF2, -CH=CHCN, -CH2CH=CH2, -CH2CH=CHCN, -CH2CH=CF2, -CH2CF=CF2, -C(CH3)=CH2, -C(CF3)=CH2, -C(CH3)=CF2, -CH=CHCF3, -CH=CHCH2O-CH3, -C(CH3)=CHCF3, -CH2CH=CHCF3, -C≡C(CF3), -CH2C≡C(CF3),

[0057] As used herein, the term "cycloalkyl" means a monocyclic, fused polycyclic, bridged polycyclic or spirocyclic saturated monovalent hydrocarbon ring structure having a specified number of ring carbon atoms. The cycloalkyl can have 3 to 12 carbon atoms (i.e., C 3-12 cycloalkyl), for example 3 to 10, 3 to 8, 3 to 7, 3 to 6, 3 to 4, 5 to 6 carbon atoms. Examples of suitable cycloalkyls include, but are not limited to, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; or polycyclic (e.g., bicyclic) structures, including spiro, fused or bridged systems, such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, spiro[3.4]octyl, bicyclo[3.1.1]hexyl, bicyclo[3.1.1]heptyl or bicyclo[3.2.1]octyl, etc. The term "C 3-6 cycloalkyl" or "C 3-4 cycloalkyl" used in the definitions of the compounds herein refers to the monocyclic cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0058] Unless otherwise defined, the "cycloalkyl" group as a substituent in the definitions of the compounds herein, such as -C 3-6 cycloalkyl or spiro-C 3-4 cycloalkyl or the "cycloalkyl" group as part of a substituent, such as -(CH2) n -C 3-6 cycloalkyl in cycloalkyl, is optionally substituted, and the substituents can be selected from one or more of the following: D, OH, NH2, halogen, CN, -C 1-6 alkyl optionally substituted with halogen or D, -O-C 1-6 alkyl optionally substituted with halogen or D, -O-CON(H or -C1-6 alkyl)2, wherein the alkyl is further optionally substituted by halogen or D. Optionally substituted -C 3-6 cycloalkyl, -(CH2) n -C 3-6 Examples of cycloalkyl include, but are not limited to:

[0059]

[0060]

[0061] where * represents the atom in the spirocycloalkyl that is connected to the rest of the molecule.

[0062] As used herein, the term "heterocycloalkyl" means a monocyclic, fused polycyclic, spirocyclic or bridged polycyclic non-aromatic saturated or unsaturated ring structure containing one or more (e.g., 1, 2, 3 or 4) heteroatoms independently selected from O, N, P, Se and S, and a specified number of ring atoms, or an N-oxide thereof, or an S-oxide or S-dioxide thereof. The heterocycloalkyl can have 3 to 12 ring members (which can be referred to as 3- to 12-membered heterocyclic groups), e.g., 3 to 10 ring members, 3 to 8 ring members, 3 to 7 ring members, 4 to 7 ring members, 4 to 6 ring members, 5 to 7 ring members, 5 to 6 ring members, 6 to 10 ring members, 6 to 12 ring members, e.g., a 4- to 7-membered monocyclic heterocycloalkyl such as a 4- to 7-membered monocyclic saturated heterocycloalkyl, a 4- to 7-membered monocyclic unsaturated heterocycloalkyl; or a 6- to 12-membered polycyclic heterocycloalkyl, such as a 6- to 10-membered spiroheterocycloalkyl, a 6- to 10-membered fused heterocycloalkyl and a 6- to 10-membered bridged heterocycloalkyl. The heterocycloalkyl typically contains at least 1 and at most 4 (e.g., 1, 2, 3 or 4) heteroatoms, e.g., a 4- to 7-membered monocyclic heterocycloalkyl such as a 4- to 7-membered monocyclic saturated heterocycloalkyl or a 4- to 7-membered monocyclic unsaturated heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, P, Se and S (preferably O, N, S), e.g., a 4- to 7-membered monocyclic saturated heterocyclic group or a bridged heterocyclic group containing 1 or 2 N atoms, or a 6- to 12-membered polycyclic heterocycloalkyl (preferably a 6- to 10-membered polycyclic heterocycloalkyl) containing 1 to 4 (preferably 1 to 3, more preferably 1 to 2) heteroatoms independently selected from N, O, P, Se and S (preferably O, N, S), e.g., a 6- to 10-membered spiroheterocycloalkyl, a fused heterocycloalkyl or a bridged heterocycloalkyl containing 1-3 N atoms and 0-1 O atoms. For example, the heterocycloalkyl of these examples can be saturated or unsaturated.Examples of suitable heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, dihydropyrrolyl, pyrrolidinyl (e.g., 1-pyrrolidinyl, 2-pyrrolidinyl, and 3-pyrrolidinyl), dihydropyrazolyl, pyrazolidinyl, dihydroimidazolyl, imidazolidinyl, partially or fully hydrogenated forms of triazolyl, partially or fully hydrogenated forms of tetrazole, partially or fully hydrogenated forms of furyl such as dihydrofuryl, tetrahydrofuryl (e.g., 1-tetrahydrofuryl, 2-tetrahydrofuryl, and 3-tetrahydrofuryl), partially or fully hydrogenated forms of thienyl such as dihydrothienyl, tetrahydrothienyl (e.g., 1-tetrahydrothienyl, 2-tetrahydrothienyl, and 3-tetrahydrothienyl), partially or fully hydrogenated forms of thiazolyl or isothiazolyl such as dihydrothiazolyl, thiazolidinyl, thiadiazolyl or its partially or fully hydrogenated forms, partially or fully hydrogenated forms of oxazolyl or isoxazolyl such as dihydrooxazolyl, oxazolidinyl, oxadiazolyl or dioxazolyl in its partially or fully hydrogenated forms such as, partially or fully hydrogenated forms of pyridyl such as dihydropyridyl, piperidinyl (e.g., 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, and 4-piperidinyl), partially or fully hydrogenated forms of pyranyl such as dihydropyranyl, tetrahydropyranyl (e.g., 4-tetrahydropyranyl), dihydrothiopyranyl, tetrahydrothiopyranyl (e.g., 4-tetrahydrothiopyranyl), morpholinyl (e.g., morpholino), thiomorpholinyl, dioxinyl, dioxenyl, dioxanyl, piperazinyl, aza-. Partially or fully hydrogenated forms of the group such as azepanyl, diaza- Partially or fully hydrogenated forms of the group such as diazepanyl e.g., 1,4-diazepanyl, oxaaza- Partially or fully hydrogenated forms of the group such as oxaazepane, 3,6-diaza-bicyclo[3.1.1]heptyl or 3-aza-bicyclo[3.2.1]octyl, Partially or fully hydrogenated forms of indolyl or isoindolyl, etc. The atom in the heterocycloalkyl group that is connected to the rest of the compound can be a carbon atom or a heteroatom, as long as it is chemically feasible. As long as the ring connected to the rest of the molecule is a non-aromatic saturated or unsaturated ring, even if the ring fused to this ring is aromatic, this fused ring is within the scope of "heterocycloalkyl" in this article. In a preferred embodiment of the present invention, the heterocycloalkyl group is saturated.

[0063] As used herein, the term "saturated" when defining a cyclic group means a monocyclic or polycyclic saturated ring containing at least one (preferably 1 to 4, more preferably 1 to 3) heteroatoms selected from N, O, S, P, and Se. Examples include aziridinyl, azetidinyl, oxetanyl, imidazolidinyl, morpholinyl, pyrrolidinyl, piperidinyl, piperazinyl, tetrahydrofuryl, tetrahydro-2H-pyranyl, tetrahydrothienyl, thiazolidinyl, oxazolidinyl, etc.

[0064] As used herein, the term "unsaturated" when defining a cyclic group means a cyclic group in which a monocyclic or polycyclic non-aromatic moiety is unsaturated. Examples of unsaturated rings include, but are not limited to, partially hydrogenated forms of the following aromatic rings: imidazolyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxadiazolyl, pyrazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyridazinyl, indolyl, isoindolyl, indazolyl, triazolo-pyridyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzothienyl, furyl, benzofuryl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl, methylenedioxyphenyl, ethylenedioxyphenyl, dihydrobenzofuryl, 1,2,3,4-tetrahydroisoquinolinyl, and the like.

[0065] As used herein, the term "hydroxyl" refers to the -OH group.

[0066] As used herein, the term "cyano" refers to the -CN group.

[0067] As used herein, the term "amino" refers to -NH2.

[0068] There are also substituted amino groups in the definition of the compounds herein, such as -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, wherein the -C 1-6 alkyl may be further optionally substituted as shown under the definition of each group. In some embodiments, the substituted amino group is -N(R b )2 in -CON(R b )2, wherein R b is -C 1-6 alkyl optionally substituted by halogen. Examples of substituted amino groups include, but are not limited to, -NH2, -NH-CH3, -NH-CH2CH3, -NH-CF3, -NH-CH2CF3, -NH-CH2CN, -NH-CH2CH2CN, -N(CH3)2, -N(CH3)(CH2CH3), -N(CH3)(CF3), -N(CH3)(CH2CF3), -N(CH3)(CH2CH2CN), -NHCH2-OCH3, N(CH3)(CH2CH2-OCH3).

[0069] As used herein, the term "optionally substituted", unless otherwise indicated, means that the group may be unsubstituted or substituted by one or more (e.g., 1, 2, 3, 4, or 5 or more, or any range derivable therefrom) of the substituents listed for the group, where when multiple substituents are present, each substituent may be the same or different. In one embodiment, the optionally substituted group has 1 substituent. In another embodiment, the optionally substituted group has 2 identical or different substituents. In another embodiment, the optionally substituted group has 3 identical or different substituents. In another embodiment, the optionally substituted group has 4 identical or different substituents. In another embodiment, the optionally substituted group has 5 identical or different substituents.

[0070] In the definitions of the compounds herein, the H carried by a saturated carbon atom may not be shown, and those skilled in the art can easily determine the number of H or non-H substituents carried by the target atom. For example, in the structural fragment when m is 1 and R3 is not H, there is an unshown H on the ring carbon atom to which it is attached, or when R3 is =C(R c )2, there is no H; when m is 2 and R3 is not H, the ring carbon atom to which it is attached does not carry an H atom, and the two R3 groups it carries can be attached to the shown ring carbon atom by single bonds respectively, or together form a spiro ring with the shown carbon atom.

[0071] In the definitions of the compounds herein, the CH2 or NH defined for a ring atom is not limited to the unsubstituted state, and its actual existence form is determined according to the overall definition of the ring group. For example, in the structural fragment , X may be CH2, but according to the overall definition of this fragment, it also includes cases where, for example, both R8 and R8' are attached to X which is CH2 and together form a spiro ring, cases where one of R8 and R8' is attached to X which is CH2, the other is attached to an adjacent ring atom and they together form a fused ring, cases where one of R8 and R8' is attached to X which is CH2, the other is attached to a non-adjacent ring atom and they together form an in-ring bridge, and cases where it is substituted by the substituents defined for this fragment. Again, for example, in the structural fragment , each G variable is defined as being able to be CH2 or NH. According to the overall definition of this fragment, this includes cases where neither of them is substituted, as well as cases where CH2 or NH is each substituted by the defined substituents, such as

[0072] Many of the groups defined herein are optionally substituted, and the list of substituents given in this definition part is merely exemplary and is not intended to limit the substituents defined in other parts of the specification and claims.

[0073] Unless otherwise specified, the C in the definitions of the compounds of the present inventionn-n+m or C n -C m covering various cases of from n to n + m carbons, such as C 1-6 including C1, C2, C3, C4, C5 and C6, and also including any range within n to n + m, such as C 0-6 including C1, C2, C3, C4, C5, C6, C 0-1 , C 0-2 , C 0-3 , C 0-4 , C 0-5 , C 1-2 , C 1-3 , C 1-4 , C 2-3 etc., C 1-6 including C 1-2 , C 1-3 , C 1-4 , C 2-6 , C 3-6 etc. Similarly, in the definition of the compounds of the present invention, n - membered to n + m - membered means that the number of ring atoms is from n to n + m. For example, a 3 - to 12 - membered ring includes a 3 - membered ring, a 4 - membered ring, a 5 - membered ring, a 6 - membered ring, a 12 - membered ring, etc., and also includes any range within n to n + m. For example, a 3 - to 12 - membered ring includes a 3 - to 6 - membered ring, a 3 - to 8 - membered ring, a 3 - to 9 - membered ring, a 4 - to 10 - membered ring, a 4 - to 7 - membered ring, a 4 - to 5 - membered ring, a 5 - to 6 - membered ring, a 5 - to 7 - membered ring, a 5 - to 8 - membered ring, a 5 - to 9 - membered ring, a 6 - to 7 - membered ring, a 6 - to 8 - membered ring, a 6 - to 10 - membered ring, and a 6 - to 12 - membered ring, etc.

[0074] Those of ordinary skill in the art of organic synthesis understand that for each group carried on the structure of the compounds of the present invention, whether unsubstituted or substituted by various defined substituents, it is on the premise that the compound molecule is chemically feasible and stable, and the type and number of the substituents are determined by the number and valence of the atoms in the group.

[0075] As used in this specification and the following claims, the word "comprising" and variations of this word such as "including" and "containing" mean "including but not limited to", and are not intended to exclude, for example, other additives, ingredients, integers or steps. When an element is described as including a plurality of components, steps or conditions, it should be understood that the element can also be described as including any combination of the plurality of components, steps or conditions, or "consisting of a plurality of or combined components, steps or conditions" or "consisting essentially of a plurality of or combined components, steps or conditions".

[0076] It should be understood that when the dosage involved in the description of the compounds of the present invention, pharmaceutical compositions containing the same, pharmaceutical combinations, kits and related uses and methods herein is based on the weight of the free form, excluding any salts, hydrates or solvates thereof, unless it is indicated in the specification that the dosage is based on the weight of the salt, hydrate or solvate.

[0077] Problems to be Solved by the Invention

[0078] As described above, compounds capable of inhibiting RAS proteins, especially KRAS mutant proteins (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutant proteins) and KRAS wild amplified cells can be used for the treatment or prevention of diseases mediated by said proteins (such as cancer or tumor). Therefore, in this field, various structural types of RAS inhibitors have been developed. However, there are still problems to be solved with existing KRAS inhibitors, including, for example, the anti-tumor activity of many inhibitors is not satisfactory, or they have toxic side effects resulting in poor drug resistance, or their pharmacokinetic properties are insufficient to allow administration by a convenient route, i.e., poor "drugability", or they cause undesirable drug interactions due to the inhibitory effect on the cytochrome P450 enzyme system, and so on. Further, even for inhibitors with good anti-tumor activity, it is still desirable to further improve their selective inhibitory activity against the target protein in vivo, further improve their drug resistance (less toxic side effects or better safety) and further improve their pharmacokinetic properties through structural optimization, in order to provide more and better treatment options clinically.

[0079] Methods for Solving the Problems

[0080] Through extensive and in-depth research, the present inventors have developed a group of compounds that have significant inhibitory activity against RAS proteins, especially KRAS mutant proteins (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutant proteins) and KRAS amplified cells. Through structural modification and activity verification, the present inventors found that by carrying out specific types of substituent modifications at several specific positions of the benzopyrimidine ring and quinazoline in the structure of the KRAS inhibitor, a specific combination of several substitution sites and substituent types was obtained, which achieved further improved inhibitory activity against KRAS mutant proteins compared with the prior art inhibitors. Moreover, the compounds thus modified have good safety, a reduced risk of drug interactions, and also have good, even further improved, pharmacokinetic properties, enabling administration by a convenient route.

[0081] The present invention mainly provides effective RAS inhibitors, specifically KRAS inhibitors (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification inhibitors) compounds; pharmaceutical compositions containing such compounds as active ingredients; the compounds as drugs for treating or preventing tumors or cancers mediated by RAS, specifically KRAS (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification) or benefited from the inhibition of RAS, specifically KRAS (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification); methods of using the compounds for treating or preventing diseases such as tumors or cancers mediated by RAS, specifically KRAS (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification) or benefited from the inhibition of RAS, specifically KRAS (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification); and the use of the compounds in the preparation of drugs for treating or preventing diseases such as tumors or cancers mediated by RAS, specifically KRAS (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification) or benefited from the inhibition of RAS, specifically KRAS (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification).

[0082] The present invention thus provides the following technical solutions.

[0083] Compounds of the Invention

[0084] Terms such as "compounds of the invention" and "compounds of the present invention" used throughout this application, unless otherwise defined, cover the compounds defined in each embodiment and its preferred embodiments herein, or their respective specific embodiments, including their isomers, including atropisomers, enantiomeric mixtures, especially racemates, diastereomeric mixtures, geometric isomers, tautomers, solvates, metabolites, prodrugs, isotopic variants and salts (such as pharmaceutically acceptable salts).

[0085] Accordingly, the above-mentioned various isomers and derivatives of the compounds of the present invention are hereby all covered within the scope of the present invention, with their respective meanings, preparations and specific examples as defined in the "Definitions" section above or well-known to those skilled in the art. However, preferably they are the compounds of the present invention and / or their pharmaceutically acceptable salts or solvates.

[0086] The present invention also covers the N-oxides of the compounds of the present invention, provided that these compounds contain a basic nitrogen atom such as the nitrogen atom present in a nitrogen-containing heterocycle and are chemically and biologically viable. Certain compounds of the present invention may exist in polymorphic or amorphous forms, and thus they also fall within the scope of the present invention.

[0087] In a first aspect, the present invention provides the following compound embodiments.

[0088] Embodiment 1: A compound of formula (I), its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates,

[0089]

[0090] wherein:

[0091] M is selected from N or C-R9;

[0092] W is selected from N or C-R 10 ;

[0093] R9 is selected from H, halogen, CN, NO2 and -C 1-6 alkyl optionally substituted with halogen;

[0094] R 10 is selected from H, halogen, CN, OH, -C 1-6 alkyl optionally substituted with halogen and -OC 1-6 alkyl optionally substituted with halogen;

[0095] Z is selected from H, OH and NH2;

[0096] X is selected from CH2 and O, provided that when k is 0, X is CH2;

[0097] Y is selected from O, S, Se and N-R a ;

[0098] R1 is selected from H and -C 1-6 alkyl optionally substituted, wherein the substituents are selected from halogen, D and -O-C 1-6 alkyl optionally substituted with halogen or D;

[0099] Each R2 is independently selected from H, D and -C 1-6 alkyl optionally substituted, wherein the substituents are selected from halogen, D and -O-C1-6 alkyl;

[0100] R3 is selected from H, D, halogen, -CN, -OH, -NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -O-C 1-6 alkyl, -O-C 3-6 cycloalkyl, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl, -(CH2) n -C 3-6 cycloalkyl and =C(R c )2, where each occurrence of C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl is optionally substituted, and the substituents are selected from halogen, CN, D, and -OC 1-6 alkyl optionally substituted by halogen, where the C 3-6 cycloalkyl is optionally substituted, and the substituents are selected from halogen, CN, D, -C 1-6 alkyl optionally substituted by halogen, and -OC 1-6 alkyl optionally substituted by halogen, or

[0101] two R3 groups attached to the same ring carbon atom together with the carbon atom to which they are attached form a spiro C 3-6 cycloalkyl or a spiro 4-7-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, S, and the cycloalkyl or heterocycloalkyl is optionally substituted by halogen or -C 1-6 alkyl optionally substituted by halogen;

[0102] R4 is selected from H, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl, and -(CH2) n -C 3-6 cycloalkyl, where -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl is optionally substituted, and the substituents are selected from D, halogen, CN, OH, -O-C 1-6 alkyl optionally substituted by halogen or D, and -OCON(R b )2, where the C 3-6 cycloalkyl is optionally substituted, and the substituents are selected from D, halogen, CN, OH, -C 1-6 alkyl optionally substituted by halogen or D, -O-C 1-6 alkyl optionally substituted by halogen or D, and -OCON(R b )2;

[0103] R5 is selected from H, halogen, -CN, -NO2;

[0104] R6 is selected from halogen, CN, -C 1-6 alkyl and -C 2-6 alkynyl, wherein the -C 1-6 alkyl and -C 2-6 alkynyl are each independently optionally substituted with halogen;

[0105] R7 is selected from H, halogen, CN, -C 1-6 alkyl optionally substituted with halogen or D, -OC 1-6 alkyl optionally substituted with halogen or D, and -C 2-6 alkynyl optionally substituted with halogen or D;

[0106] R8 and R8' attached to non-adjacent ring carbon atoms together form a bridged ring within the ring -(CH2) 1-2 - or -CH2=CH2-,

[0107] or R8 and R8' attached to the same ring carbon atom together with the ring carbon atom to which they are attached form a 4-6 membered spiroalkyl ring or a 4-6 membered spiroheteroalkyl ring containing 1 to 3 heteroatoms independently selected from N, O, and S,

[0108] or R8 and R8' attached to adjacent ring carbon atoms together with the ring carbon atoms to which they are attached form a fused C 3-6 cycloalkyl or a 4-6 membered fused heterocyclic ring containing 1 to 2 heteroatoms independently selected from N, O, and S,

[0109] wherein the bridged ring, spiro ring, or fused ring is each independently optionally substituted, and the substituents are selected from OH, oxo, -OC 1-6 alkyl optionally substituted with halogen, and -C 1-6 alkyl optionally substituted with halogen;

[0110] R a and R b are each independently selected from H and -C 1-6 alkyl optionally substituted with halogen;

[0111] R c are each independently selected from H, halogen, and -C 1-6 alkyl optionally substituted with halogen;

[0112] k and n are each independently selected from integers from 0 to 3; and

[0113] m is selected from integers from 0 to 6.

[0114] Embodiment 1.1: A compound of formula (I) according to Embodiment 1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein M is N.

[0115] Embodiment 1.2: A compound of formula (I) according to Embodiment 1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein M is C-R9.

[0116] Embodiment 1.2.1: A compound of formula (I) according to Embodiment 1.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R9 is H.

[0117] Embodiment 1.2.2: A compound of formula (I) according to Embodiment 1.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R9 is a halogen, such as F, Cl, Br, I.

[0118] Embodiment 1.2.3: A compound of formula (I) according to Embodiment 2.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R9 is CN; or R9 is NO2.

[0119] Embodiment 1.2.4: A compound of formula (I) according to Embodiment 2.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R9 is -C 1-6 alkyl, preferably -C 1-3 alkyl, optionally substituted by a halogen, preferably substituted by a halogen, more preferably substituted by F.

[0120] Embodiment 1.3: A compound of formula (I) according to Embodiment 1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein M is selected from any one or any combination of Embodiments 1.1 to 1.2.4; for example, M is selected from N, C-halogen (such as C-F, C-Cl), C-CN, C-NO2, C-halogen-substituted C 1-6 alkyl (such as C-CF3); preferably M is N.

[0121] Embodiment 2.1: A compound of formula (I) according to any one of Embodiments 1 to 1.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein W is N.

[0122] Embodiment 2.2: A compound of formula (I) according to any one of Embodiments 1 to 1.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein W is C-R 10 。

[0123] Embodiment 2.2.1: A compound of formula (I) according to Embodiment 2.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R 10 is H.

[0124] Embodiment 2.2.2: A compound of formula (I) according to Embodiment 2.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R 10 is a halogen, such as F, Cl, Br, I, preferably F; or R 10 is CN.

[0125] Embodiment 2.2.3: A compound of formula (I) according to Embodiment 2.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R 10 is OH.

[0126] Embodiment 2.2.4: A compound of formula (I) according to Embodiment 2.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R 10 is -C 1-6 alkyl, preferably -C 1-3 alkyl, optionally substituted by a halogen.

[0127] Embodiment 2.2.5: A compound of formula (I) according to Embodiment 2.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R 10 is -OC 1-6 alkyl, preferably -OC 1-3 alkyl, wherein the alkyl is optionally substituted by a halogen.

[0128] Embodiment 2.3: A compound of formula (I) according to any one of Embodiments 1 to 1.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein W is selected from any one of Embodiments 2.1 to 2.2.5 or any combination thereof; for example, W is selected from N, C-halogen, C-CN, C-halogen-substituted -C 1-6 alkyl, C-halogen-substituted -OC 1-6 alkyl, such as N, C-F, C-Cl, C-CN, C-CF3; preferably W is C-halogen, more preferably C-F.

[0129] Embodiment 3.1: A compound of formula (I) according to Embodiment 1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the fused parent ring containing M and W includes but is not limited to: wherein R9 is as defined in any one of Embodiments 1.2.1 to 1.3, R 10 as defined in any one of Embodiments 2.2.1 to 2.3, for example but not limited to:

[0130]

[0131] Embodiment 4.1: A compound of formula (I) according to any one of Embodiments 1 to 3.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R7 is H.

[0132] Embodiment 4.2: A compound of formula (I) according to any one of Embodiments 1 to 3.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R7 is halogen; or R7 is CN.

[0133] Embodiment 4.3: A compound of formula (I) according to any one of Embodiments 1 to 3.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R7 is -C 1-6 alkyl optionally substituted by halogen or D, preferably -C 1-3 alkyl, such as -CH3, -CD3.

[0134] Embodiment 4.4: A compound of formula (I) according to any one of Embodiments 1 to 3.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R7 is -OC 1-6 alkyl optionally substituted by halogen or D, preferably -OC 1-3 alkyl, such as -O-CH3, -O-CD3; in a specific embodiment, at this time M is N.

[0135] Embodiment 4.5: A compound of formula (I) according to any one of Embodiments 1 to 3.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R7 is -C 2-6 alkynyl optionally substituted by halogen or D, preferably -C 2-4 alkynyl, such as but not limited to ethynyl, prop-1-ynyl.

[0136] Embodiment 4.6: A compound of formula (I) according to any one of Embodiments 1 to 3.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R7 is selected from any one of Embodiments 4.1 to 4.5 or any combination thereof, for example, R7 is selected from H, -OC optionally substituted by halogen or D 1-6 alkyl and -C optionally substituted by halogen or D 2-6 alkynyl, for example, R7 is selected from H and -OC optionally substituted by halogen or D 1-6 alkyl, more specifically selected from H and -OC optionally substituted by D 1-6 alkyl.

[0137] Embodiment 5.1: A compound of formula (I) according to any one of Embodiments 1 to 4.6, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R5 is H.

[0138] Embodiment 5.2: A compound of formula (I) according to any one of Embodiments 1 to 4.6, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R5 is halogen, preferably F.

[0139] Embodiment 5.3: A compound of formula (I) according to any one of Embodiments 1 to 4.6, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R5 is -CN; or R5 is -NO2.

[0140] Embodiment 5.4: A compound of formula (I) according to any one of Embodiments 1 to 4.6, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R5 is selected from any one of Embodiments 5.1 to 5.3 or any combination thereof; for example, R5 is selected from H and halogen (preferably F), for example, R5 is selected from halogen (preferably F).

[0141] Embodiment 6.1: A compound of formula (I) according to any one of Embodiments 1 to 5.4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R6 is halogen, such as F or Cl.

[0142] Embodiment 6.2: A compound of formula (I) according to any one of Embodiments 1 to 5.4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R6 is CN.

[0143] Embodiment 6.3: A compound of formula (I) according to any one of Embodiments 1 to 5.4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R6 is -C optionally substituted by halogen1-6 Alkyl, preferably -C optionally substituted by halogen 1-3 Alkyl, such as -C 1-3 Alkyl, such as ethyl.

[0144] Embodiment 6.4: A compound of formula (I) according to any one of Embodiments 1 to 5.4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R6 is -C optionally substituted by halogen 2-6 Alkynyl, preferably -C optionally substituted by halogen 2-4 Alkynyl, such as ethynyl.

[0145] Embodiment 6.5: A compound of formula (I) according to any one of Embodiments 1 to 5.4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R6 is selected from any one of Embodiments 6.1 to 6.4 or any combination thereof, for example, R6 is selected from halogen, -C optionally substituted by halogen 1-6 Alkyl and -C optionally substituted by halogen 2-6 Alkynyl, for example, R6 is selected from halogen, -C 1-3 Alkyl and -C 2-4 Alkynyl, such as F, Cl, ethyl and ethynyl.

[0146] Embodiment 7.1: A compound of formula (I) according to any one of Embodiments 1 to 6.5, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein Z is H; in a specific embodiment, Z is H, R5 is H and R6 is halogen, such as F or Cl.

[0147] Embodiment 7.2: A compound of formula (I) according to any one of Embodiments 1 to 6.5, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein Z is OH, or Z is NH 2 ; in a specific embodiment, Z is OH, R5 is selected from halogen, and R6 is selected from halogen, -C optionally substituted by halogen 1-6 Alkyl and -C optionally substituted by halogen 2-6 Alkynyl, for example, R6 is selected from halogen, -C 1-3 Alkyl and -C 2-4 Alkynyl.

[0148] Embodiment 8: A compound of formula (I) according to any one of Embodiments 1 to 7.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein in the structural fragment , R5 is selected from H and halogen, preferably halogen (more preferably F); R6 is selected from halogen, -C optionally substituted by halogen 1-6Alkyl and optionally halogen-substituted -C 2-6 alkynyl, preferably R6 is selected from halogen, -C 1-3 alkyl and -C 2-4 alkynyl; Z is selected from H and OH, preferably OH; specific examples include but are not limited to:

[0149] Embodiment 9.1: A compound of formula (I) according to any one of Embodiments 1 to 8, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein in the structural fragment when,

[0150] when k is 0, the fragment is an azetidine having an internal bridging -(CH2) 1-2 - or -CH2=CH2-, azetidine spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O and S), or azetidine fused (C 3-6 cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O and S);

[0151] when k is 1, the fragment is a pyrrolidine or oxolane having an internal bridging -(CH2) 1-2 - or -CH2=CH2-, (pyrrolidine or oxolane) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O and S), or (pyrrolidine or oxolane) fused (C 3-6 cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O and S);

[0152] when k is 2, the fragment is a piperidine or morpholine having an internal bridging -(CH2) 1-2 - or -CH2=CH2-, (piperidine or morpholine) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O and S), or (piperidine or morpholine) fused (C 3-6 cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O and S);

[0153] when k is 3, the fragment is a heptamethyleneimine or oxepane having an internal bridging -(CH2) 1-2 - or -CH2=CH2-, (heptamethyleneimine or oxepane) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O and S), or (heptamethyleneimine or oxepane) fused (C 3-6cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O, and S);

[0154] wherein the shared ring carbon atom of the above spiro ring can be in the meta or para position of the nitrogen heteroatom, preferably the meta position;

[0155] wherein the above bridged ring, spiro ring or fused ring is each independently optionally substituted, and the substituents are selected from OH, oxo, -OC 1-6 alkyl optionally substituted by halogen and -C 1-6 alkyl optionally substituted by halogen, for example selected from OH and oxo;

[0156] Preferably, the structural fragment is the above spiro ring or fused ring structure.

[0157] Embodiment 9.2: A compound of formula (I) according to any one of Embodiments 1 to 9.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment is i.e., k is 0; i.e., k is 1; i.e., k is 2 or 3.

[0158] Embodiment 9.2.1: A compound of formula (I) according to Embodiment 9.1 or 9.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment is azetidine spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, and S), (azetidine or oxazetidine) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, and S), (piperidine or piperidine N-oxide) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, and S) or (azepane or azepane N-oxide) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, and S); preferably (piperidine) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, and S); the ring carbon atom shared by the spiro ring can be in the para or meta position of the nitrogen heteroatom connected to the rest of the molecule as appropriate; for example but not limited to

[0159] In a specific embodiment, R8 and R8' form a spiro 4-6-membered cycloalkyl or a spiro 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, and S with the ring carbon atom in the meta position of the N heteroatom of piperidine, that is, represented as wherein G1 is selected from CH2, NH, O, and S, at least one of G2, G3, and G4 is selected from CH2, NH, O, and S, and the rest are each independently optionally selected from absent, CH2, NH, O, and S;

[0160] In a more specific embodiment, G3 and G4 in are absent, i.e., wherein both G1 and G2 are CH2, or one of them is CH2 and the other is selected from NH, O, and S (preferably NH and O); the common ring carbon atoms marked with an asterisk may optionally have chirality, and accordingly, this fragment may be in a racemic form or there exist

[0161] In a more specific embodiment, G4 in is absent, i.e.,

[0162] wherein G1, G2, and G3 are each independently selected from CH2, NH, O, and S; 1-6 1-6 1-6 The spiro ring in each of the above embodiments is optionally substituted at any chemically feasible position, for example, substituted on the spiro ring formed by R8 and R8' together with the ring carbon atoms to which they are attached, and the substituents are selected from OH, oxo, -OC

[0163] alkyl optionally substituted with halogen and -C

[0164] Embodiment 9.2.2: A compound of formula (I) according to Embodiment 9.1 or 9.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment is azetidine-fused (C 3-6 cycloalkyl or a 4-6 membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O, and S), (pyrrolidine or pyrroline) -fused (C 3-6 cycloalkyl or a 4-6 membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O, and S), (piperidine or piperidine N-oxide) -fused (C 3-6 cycloalkyl or a 4-6 membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O, and S) or (azepane or azepane N-oxide) -fused (C 3-6 cycloalkyl or a 4-6 membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O, and S); preferably (azepane or azepane N-oxide) -fused (C 3-6cycloalkyl or a 4-6 membered heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O and S); more preferably (azepane) fused (C 3-6 cycloalkyl); such as but not limited to

[0165] In one specific embodiment, the ring containing N and X is azepane or azepanone, and R8 and R8' attached to adjacent ring carbon atoms together with the atoms to which they are attached form C 3-6 cycloalkyl;

[0166] One specific embodiment is More specifically May be present as appropriate in Present, preferably

[0167] The fused rings in each of the above embodiments are optionally substituted at any chemically feasible position, and the substituents are selected from OH, oxo, -OC 1-6 alkyl optionally substituted with halogen and -C 1-6 alkyl optionally substituted with halogen; preferably unsubstituted.

[0168] Embodiment 9.2.3: A compound of formula (I) according to Embodiment 9.1 or 9.2, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein in the structural fragment k is an integer from 1 to 3, preferably 2 to 3, X is selected from CH2 and O, preferably selected from CH2, R8 and R8' are attached to non-adjacent carbon atoms and together form an in-ring bridging -(CH2) 1-2 - or -CH2=CH2-, preferably -(CH2) 1-2 -; Specific examples include but are not limited to The bridged ring is optionally substituted with OH, oxo, -OC 1-6 alkyl optionally substituted with halogen and -C 1-6 alkyl, such as substituted with OH or -OC 1-6 alkyl optionally substituted with halogen, such as substituted with OH.

[0169] Embodiment 9.3: A compound of formula (I) according to any one of Embodiments 1 to 9.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment As defined in any one of embodiments 9.2 to 9.2.2 or any combination thereof, for example selected from optionally substituted (azacyclohexane) spiro(4-6-membered cycloalkyl or 4-6-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O and S) and (oxazepane) fused (C 3-6 cycloalkyl), specifically as defined above Or as defined in embodiment 9.2.3, for example having an internal bridging -(CH2) 1-2 - or -CH2=CH2-, preferably -(CH2) 1-2 - of an optionally substituted 5- to 7-membered heterocycloalkyl.

[0170] Embodiment 10.1: A compound of formula (I) according to any one of embodiments 1 to 9.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein Y is O.

[0171] Embodiment 10.2: A compound of formula (I) according to any one of embodiments 1 to 9.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein Y is S.

[0172] Embodiment 10.3: A compound of formula (I) according to any one of embodiments 1 to 9.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein Y is Se.

[0173] Embodiment 10.4: A compound of formula (I) according to any one of embodiments 1 to 9.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein Y is N-R a , R a is H; or Y is N-R a , R a is optionally halogen-substituted -C 1-6 alkyl, preferably optionally halogen-substituted -C 1-3 alkyl.

[0174] Embodiment 11.1: A compound of formula (I) according to any one of embodiments 1 to 10.4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment is preferably more preferably wherein the ring carbon atom connecting R3 may optionally have

[0175] chirality, whereby R3 has an R or S stereoconfiguration.

[0176] Embodiment 11.2: The compound of formula (I) according to Embodiment 11.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R2 is selected from H or D, for example, both R2 are H; or both R2 are D; or one of R2 is H and the other is D.

[0177] Embodiment 11.2.1: The compound of formula (I) according to Embodiment 11.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein one R2 is H or D, and the other R2 is -C 1-6 alkyl (preferably -C 1-3 alkyl), optionally substituted by halogen, D, -O-C 1-6 alkyl substituted by halogen or D, or both R2 are independently -C 1-6 alkyl (preferably -C 1-3 alkyl), optionally substituted by halogen, D, -O-C 1-6 alkyl substituted by halogen or D.

[0178] Embodiment 11.3: The compound of formula (I) according to any one of Embodiments 11.1 to 11.2.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R1 is H; or R1 is -C 1-6 alkyl, preferably -C 1-3 alkyl, optionally substituted by halogen, D, or -O-C 1-6 alkyl substituted by halogen or D; preferably R1 is -C 1-3 alkyl or deuterated C 1-3 alkyl, such as -CH3, -CH2D, -CHD2, -CD3.

[0179] Embodiment 11.4: The compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is H; or R3 is D.

[0180] Embodiment 11.4.1: The compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is halogen, such as F, Cl, Br, I, preferably F; or R3 is CN.

[0181] Embodiment 11.4.2: The compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is -OH.

[0182] Embodiment 11.4.3: A compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is -NH2, -NH-C 1-6 alkyl, -N(C 1-6 alkyl)2, wherein the -C 1-6 alkyl is optionally substituted by halogen, CN, D and -OC 1-6 alkyl optionally substituted by halogen.

[0183] Embodiment 11.4.4: A compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is -C 1-6 alkyl, preferably -C 1-3 alkyl, or is -C 2-6 alkenyl, preferably -C 2-4 alkenyl, or is -C 2-6 alkynyl, preferably -C 2-4 alkynyl; each optionally substituted by halogen, CN, D and -OC 1-6 alkyl optionally substituted by halogen, for example optionally substituted by halogen, such as substituted by halogen, such as substituted by F.

[0184] Embodiment 11.4.5: A compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is -O-C 1-6 alkyl, preferably -O-C 1-3 alkyl, wherein the alkyl is optionally substituted by halogen, CN, D and -OC 1-6 alkyl optionally substituted by halogen.

[0185] Embodiment 11.4.6: A compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is -O-C 3-6 cycloalkyl, or R3 is -(CH2) 0-3 -C 3-6 cycloalkyl, preferably -(CH2) 0-3 -C 3-4 cycloalkyl, wherein the C 3-6 cycloalkyl is each optionally substituted by halogen, CN, D, -C 1-6 alkyl optionally substituted by halogen and -OC 1-6 alkyl optionally substituted by halogen.

[0186] Embodiment 11.4.7: A compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is =C(R c )2, wherein R c are each independently selected from H, halogen, -C 1-6 alkyl optionally substituted with halogen, preferably selected from H and halogen (preferably F); R3 is, for example but not limited to, =CH2, =CHF, =CF2, =CHCl, =CCl2, =C(CH3)2, =CHCH3, =CHCF3, =C(CF3)2.

[0187] Embodiment 11.4.8: A compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein two R3s attached to the same ring carbon atom together with the ring carbon atom to which they are attached form a spiro C 3-6 cycloalkyl or a spiro 4-7-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, S, each optionally substituted with halogen and optionally substituted -C 1-6 alkyl, for example but not limited to spirocyclopropyl, spirocyclobutyl, spirocyclopentyl, spiroazetidine, spiroazepane, each optionally substituted with halogen (preferably F) or optionally substituted C 1-6 alkyl (preferably -CF3); for example but not limited to where * represents the common carbon atom of the spiro ring.

[0188] Embodiment 11.4.9: A compound of formula (I) according to any one of Embodiments 11.1 to 11.3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is selected from any one of Embodiments 11.4 to 11.4.8 or any combination thereof; for example, R3 is selected from halogen (preferably F); -C 1-6 alkyl (preferably C 1-3 alkyl), wherein the alkyl is optionally substituted with halogen, preferably substituted with halogen, more preferably substituted with F; or =C(R c )2, wherein R c are each independently selected from H, halogen (preferably F), -C 1-6 alkyl optionally substituted with halogen (preferably -C 1-3 alkyl), preferably R c is selected from H and halogen (preferably F); preferably, R3 is substituted at the para position of the ring N atom;

[0189] wherein the para ring carbon atom of the ring N atom has chirality as appropriate and can be in the R or S configuration;

[0190] wherein the para - ring carbon atom of the ring N atom is substituted with =C(R c )2, the double bond may be a cis - trans isomer as appropriate, including E or Z type, preferably E type.

[0191] Embodiment 11.5: A compound of formula (I) according to any one of Embodiments 11.4 to 11.4.9, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein m is an integer from 0 to 6, preferably an integer from 1 to 4, more preferably an integer from 1 to 2, and R3 may be selected from any of the foregoing R3 embodiments or any combination thereof;

[0192] More specifically, m is 1, and R3 is selected from C 1-3 alkyl, substituted with a halogen, preferably substituted with F; and =C(R c )2, wherein R c are each independently selected from H and halogen (preferably F).

[0193] Embodiment 11.6: A compound of formula (I) according to any one of Embodiments 11.1 to 11.5, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R4 is H.

[0194] Embodiment 11.6.1: A compound of formula (I) according to any one of Embodiments 11.1 to 11.5, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R4 is -C 1-6 alkyl, preferably -C 1-3 alkyl; or R4 is -C 2-6 alkenyl, preferably -C 2-4 alkenyl; or R4 is -C 2-6 alkynyl, preferably -C 2-4 alkynyl; each optionally substituted with D, halogen, CN, OH, -O-C 1-6 alkyl optionally substituted with halogen or D and -OCON(R b )2, preferably optionally substituted with halogen or D.

[0195] Embodiment 11.6.2: A compound of formula (I) according to any one of Embodiments 11.1 to 11.5, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R4 is -(CH2) 0-3 -C 3-6 cycloalkyl, and the -C 3-6 cycloalkyl is optionally substituted with D, halogen, CN, OH, -C 1-6 alkyl optionally substituted with halogen or D, -O-C 1-6 alkyl optionally substituted with halogen or D and -OCON(R b) Substituted with 2.

[0196] Embodiment 11.6.3: A compound of formula (I) according to any one of Embodiments 11.1 to 11.5, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R4 is selected from any one of Embodiments 11.6 to 11.6.2 or any combination thereof; for example, R4 is selected from -C 1-6 alkyl and -C 3-6 cycloalkyl, optionally substituted by D, halogen, CN, OH, -C 1-6 alkyl optionally substituted by halogen or D, -O-C 1-6 alkyl and -OCON(R b )2; preferably, R4 is selected from -C 1-3 alkyl and deuterated C 1-3 alkyl, such as, but not limited to, -CH3, -CD3, -CH2CH3, -CD2CD3.

[0197] Embodiment 11.7: A compound of formula (I) according to Embodiment 11.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein:

[0198] R3 is selected from halogen (preferably F); -C 1-6 alkyl (preferably C 1-3 alkyl), wherein the alkyl is optionally substituted by halogen, preferably substituted by halogen, more preferably substituted by F; or =C(R c )2, wherein R c are each independently selected from H, halogen (preferably F), -C 1-6 alkyl optionally substituted by halogen (preferably -C 1-3 alkyl), preferably R c is selected from H and halogen (preferably F);

[0199] Preferably:

[0200] R2 are each independently selected from H and D; R1 is selected from optionally deuterated -C 1-6 alkyl; m is 1 or 2; R3 is selected from halogen, -C 1-6 alkyl and =C(R c )2, wherein -C 1-6 alkyl is optionally substituted by halogen, wherein R c are each independently selected from H, halogen, -C 1-6 alkyl optionally substituted by halogen, preferably, R3 is substituted at the para position of the ring N atom; R4 is selected from optionally deuterated -C 1-6 alkyl; more preferably:

[0201] R2 are each independently selected from H and D;

[0202] R1 is selected from -C 1-3 alkyl, optionally substituted with one or more deuteriums;

[0203] m is 1 or 2, such as 1;

[0204] R3 is substituted at the para position of the ring N atom and is selected from halogen, -C 1-3 alkyl substituted with halogen (preferably F) and =C(R c )2, where R c are each independently selected from H, halogen, -C 1-3 alkyl substituted with halogen. Specific examples of R3 include, but are not limited to, fluorine, difluorine, fluoromethyl, difluoromethyl, fluoromethylene, difluoromethylene;

[0205] R4 is selected from -C 1-3 alkyl, optionally substituted with one or more deuteriums, such as -CH3, -CH2CH3, -CD3, -CD2CD3; where the para ring carbon atom of the ring N atom to which R3 is attached is chiral as appropriate and can be in the R or S configuration.

[0206] Embodiment 11.8: A compound of formula (I) according to Embodiment 11.1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein examples of the structural fragment include, but are not limited to

[0207]

[0208]

[0209]

[0210] Preferably

[0211]

[0212] More preferably

[0213] Embodiment 12: A compound of formula (I) according to Embodiment 1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, which have the following sub-formula:

[0214]

[0215]

[0216]

[0217] Each of the substituents in each of the sub-generic formulas and their examples has the general or preferred meaning or any combination thereof defined in the corresponding foregoing embodiments; each sub-generic formula also encompasses compounds formed by any combination of the general or preferred meaning of each substituent therein with the general or preferred meaning of the remaining substituents;

[0218] In a set of embodiments, in each of the above sub-generic formulas and their examples, M is N, W is C-halogen, preferably C-F;

[0219] In a set of embodiments, in each of the above sub-generic formulas and their examples, M is selected from C-halogen (such as C-F, C-Cl), C-CN, C-NO2, C-halogen-substituted C 1-6 alkyl (such as C-CF3), and W is C-halogen, preferably C-F.

[0220] Embodiment 13: A compound selected from the compounds of the following examples, their stereoisomers, pharmaceutically acceptable salts or solvates.

[0221] It should be noted that the compounds of the present invention cover each of the above independent embodiments or each specific embodiment, also cover the embodiments constituted by any combination or sub-combination of the above embodiments or specific embodiments, and also cover the embodiments constituted by any combination of any of the above preferred or exemplified embodiments.

[0222] Advantages of the Invention

[0223] As mentioned above, it is known that RAS proteins, especially KRAS mutant proteins, play a role in tumorigenesis and a variety of other diseases. We have surprisingly found that the compounds of the present invention having the above structural characteristics can strongly inhibit cell proliferation in cell lines carrying KRAS mutant proteins (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutant protein) and KRAS wild-type amplification, thereby having potential value as anti-proliferation, pro-apoptosis and / or anti-invasive drugs in preventing, suppressing and / or treating related tumor diseases. In particular, the compounds of the present invention are expected to be useful for preventing or treating diseases or disorders mediated by or suppressed by RAS mutations, especially KRAS mutant proteins (e.g., G12C mutations, G12D mutations, G12V mutations, G12A mutations, G12R mutations, G12S mutations, G13D mutations and Q61H mutations) or KRAS wild-type amplifications, such as cancer or tumors as defined herein.

[0224] Specifically, it has been found through research that the compounds of the present invention can achieve one or more of the following technical effects:

[0225] High mutant protein inhibitory activity: The compounds of the present invention, especially the compounds specifically exemplified herein, show proliferation inhibitory activity in RAS-related cells, especially KRAS mutant cells (e.g., G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutation) and KRAS amplified cell proliferation inhibition experiments, with IC50 values ​​in the range of 0.0001-10 μM, preferably in the range of 0.0001-1 μM, as verified in Activity Examples 1, 2, 5-6;

[0226] ·Good pharmacokinetic properties, such as long t 1 / 2 , which can, for example, increase the dosing interval, extend the half-life, and enable patients to have better compliance; have the best AUC for the safety / activity combination 0-t Data, with more

[0227] Good drugability, higher bioavailability, and convenient oral administration, as verified in Active Example 3; and / or having a significantly satisfactory safety profile, a reduced risk of drug interactions, and no significant inhibitory effect on key CYP subtypes of drug metabolism, as verified in Active Example 4.

[0228] Based on the beneficial effects of the compounds of the present invention described above, the present invention also provides technical solutions in the following aspects.

[0229] Compounds of the Invention for Therapeutic Use or as Medicaments

[0230] On the one hand, the present invention provides the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, for use as a medicine.

[0231] On the other hand, the present invention provides the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, for use as RAS inhibitors, especially inhibitors of KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS wild-type amplified cells.

[0232] On the other hand, the present invention provides the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, for the treatment and / or prevention of diseases or disorders mediated by RAS proteins, specifically KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS amplification, or benefited from RAS mutations, specifically KRAS mutant proteins (such as G12C mutant, G12D mutant, G12V mutant, G12A mutant, G12R mutant, G12S mutant, G13D mutant and Q61H mutant proteins) and KRAS amplification inhibition.

[0233] In a specific embodiment, the present invention provides compounds of the present invention for treating and / or preventing diseases in which RAS proteins, specifically KRAS mutant proteins (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutant proteins) and KRAS amplification play a promoting role in the occurrence and development of the diseases, or inhibiting RAS mutant proteins, specifically KRAS mutant proteins (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutant proteins) and KRAS amplification will reduce the incidence of the diseases and reduce or eliminate the symptoms of the diseases. The diseases such as tumors or cancers include, but are not limited to: lung cancer, lung adenocarcinoma, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, melanoma of skin or intraocular, uterine cancer, ovarian cancer, rectal cancer, cancer of anal region, gastric cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor (CNS), primary CNS lymphoma, spinal tumor, brainstem glioma or pituitary adenoma.

[0234] The present invention particularly provides compounds of formula (I) or their isomers, their pharmaceutically acceptable salts or solvates that can be used to treat patients suffering from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, lung cancer, cholangiocarcinoma, endometrial cancer, ovarian cancer, leukemia; most preferably selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, cholangiocarcinoma.

[0235] Pharmaceutical Compositions and Their Administration

[0236] On the other hand, the present invention provides a pharmaceutical composition comprising the compound of formula (I) as defined above, preferably its pharmaceutically acceptable salt or solvate, and a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition of the present invention can be used to treat or prevent diseases mediated by RAS, especially KRAS mutations, such as diseases mediated by KRAS G12C, KRAS G12D, KRAS G12V, G12A, G12R, G12S or KRAS G13D mutations, or KRAS Q61H mutation, and KRAS amplification, such as tumors or cancers.

[0237] The above-mentioned pharmaceutical composition of the present invention can be formulated by techniques known to those skilled in the art, such as the techniques disclosed in Remington’s Pharmaceutical Sciences, 20th Edition. For example, it can be formulated into tablets, powders, capsules, lozenges, granules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. The composition may contain conventional components in pharmaceutical preparations, such as diluents (e.g., glucose, lactose or mannitol), carriers, pH regulators, buffers, sweeteners, fillers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, flavorants, other known additives, and other active agents. Suitable carriers and excipients are well-known to those skilled in the art and are described in detail, for example, in Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004.

[0238] The administration and application of the pharmaceutical composition of the present invention are in accordance with good medical practice. Factors to be considered in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the location of agent delivery, the method of administration, the dosing schedule, and other factors well-known to the physician practitioner. The optimal dosage level and dosing frequency of the compounds or pharmaceutical compositions of the present invention can be determined by those skilled in the art through standard tests in the field of pharmaceutical research.

[0239] The compositions of the present invention can be administered in any suitable manner, including orally, topically (including buccal and sublingual), rectally, vaginally, transdermally, parenterally, subcutaneously, intraperitoneally, intralungally, intradermally, intrathecally, by inhalation, epidurally, and intranasally, and for local treatment, also by intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. In some embodiments, the pharmaceutical composition of the present invention is administered orally.

[0240] For a 70 kg human subject, a suitable dosage range of the compounds of the present invention can be routinely determined by those skilled in the art, for example, it can be 1 - 1000 mg / day.

[0241] When the dosage of a drug or its pharmaceutically acceptable salt is described herein, it should be understood that the dosage is based on the weight of the free base and does not include any hydrates or solvates thereof, unless it is indicated in the specification that the dosage is based on the weight of the salt, hydrate or solvate.

[0242] Therapeutic Methods and Uses

[0243] As described above, the compounds of the present invention and the compounds of various specific embodiments thereof, particularly the compounds specifically prepared and characterized in the examples, exhibit inhibitory effects on RAS, particularly KRAS mutations such as KRAS G12C, KRAS G12D, KRAS G12V, G12A, G12R, G12S or KRAS G13D mutations, or KRAS Q61H, and KRAS amplified cells.

[0244] Accordingly, on the other hand, the present invention provides a method for inhibiting RAS in cells, particularly KRAS mutations, preferably KRAS G12D mutations, comprising contacting the cells with the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, to inhibit the activity of RAS mutations in cells, particularly KRAS mutations (such as G12C mutations, G12D mutations, G12V mutations, G12A mutations, G12R mutations, G12S mutations, G13D mutations and Q61H mutations) and KRAS amplification.

[0245] Based on the same property, the present invention also correspondingly provides a method for inhibiting abnormal cell growth in a mammal, comprising administering to the mammal a therapeutically effective amount of the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, or a pharmaceutical composition comprising the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates.

[0246] On the other hand, the present invention provides a method for treating and / or preventing diseases mediated by RAS, particularly KRAS mutations (such as G12C mutations, G12D mutations, G12V mutations, G12A mutations, G12R mutations, G12S mutations, G13D mutations and Q61H mutations) and KRAS amplification, comprising administering to a subject in need thereof a therapeutically effective amount of the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, or a pharmaceutical composition comprising the compounds of the present invention preferably their pharmaceutically acceptable salts or solvates.

[0247] On the other hand, the present invention provides the use of the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, or a pharmaceutical composition comprising the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, for inhibiting diseases mediated by RAS in cells, particularly KRAS mutations, preferably KRAS G12C, KRAS G12D, KRAS G12V, KRAS G12A, KRAS G12R, KRAS G12S or KRAS G13D, or KRAS Q61H mutations, and KRAS amplification.

[0248] On the other hand, the present invention provides the use of the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, or pharmaceutical compositions comprising the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, in the preparation of a medicament for the treatment and / or prevention of diseases mediated by RAS, especially KRAS mutations (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutation) and KRAS amplification.

[0249] For each of the above-described method and use technical solutions provided by the present invention, the abnormal cell growth or diseases mediated by RAS, especially KRAS mutations, preferably KRAS G12C, KRAS G12D, KRAS G12V, KRAS G12A, KRAS G12R, KRAS G12S or KRAS G13D, or KRAS Q61H mutation, and KRAS amplification especially refer to cancer or tumor. Exemplary cancers or tumors include, but are not limited to, lung cancer, lung adenocarcinoma, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, melanoma of the skin or intraocular, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal cell cancer, renal pelvic cancer, central nervous system tumors (CNS), primary CNS lymphoma, spinal tumors, brainstem glioma or pituitary adenoma.

[0250] For each of the above-described method and use technical solutions provided by the present invention, the abnormal cell growth or diseases mediated by RAS, especially KRAS mutations (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutation) and KRAS amplification are preferably selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, lung cancer, cholangiocarcinoma, endometrial cancer, ovarian cancer, leukemia; most preferably selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, cholangiocarcinoma.

[0251] Therefore, in a preferred embodiment in this regard, the present invention provides the above-described method and use technical solutions for treating or preventing cancer or tumor by inhibiting RAS mutation or amplification. In a further preferred embodiment, the present invention provides the above-described method and use technical solutions for treating or preventing pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma and cholangiocarcinoma by inhibiting RAS mutation or amplification.

[0252] The present invention also provides the use of the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, as research tool compounds for RAS inhibitors, especially KRAS inhibitors (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification inhibitors) in research. Therefore, the present invention relates to the in vitro use of the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, as RAS inhibitors, and particularly to the in vitro use of the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, as research tool compounds for the onset of RAS inhibitors. The present invention also relates to a method for inhibiting RAS, especially KRAS (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation, Q61H mutation and KRAS amplification), especially an in vitro method, which comprises administering the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, to a sample (such as a biological sample). It should be understood that the term "in vitro" is used in this specific context to mean "outside the living human body or animal", which specifically includes experiments conducted with cells, cell or subcellular extracts and / or biomolecules in an artificial environment, such as aqueous solutions or culture media that can be provided in flasks, test tubes, petri dishes, microtiter plates, etc.

[0253] Drug Combinations

[0254] The compounds of the present invention can be administered as the sole active ingredient or in combination with additional drugs or therapies.

[0255] Therefore, on the other hand, the present invention provides a pharmaceutical combination comprising the compounds of the present invention, preferably their pharmaceutically acceptable salts or solvates, and other active agents, or consisting of the two. The pharmaceutical combination is used for inhibiting abnormal cell growth in mammals, or for treating and / or preventing diseases mediated by RAS, preferably KRAS mutations (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, G13D mutation and Q61H mutation) and KRAS amplification.

[0256] The other active agents can be one or more additional compounds of the present invention, or can be a second or additional (such as a third) compound that is compatible with the compounds of the present invention, i.e., does not adversely affect each other, or has complementary activities. For example, these active agents can be compounds known to regulate other biological activity pathways, or can be compounds that regulate different components in the biological activity pathway involved in the compounds of the present invention, or even compounds that overlap with the biological targets of the compounds of the present invention.

[0257] In a specific embodiment, other active agents that can be used in combination with the compounds of the present invention include, but are not limited to, chemotherapeutic agents, therapeutic antibodies, and radiotherapy, such as alkylating agents, antimetabolites, cell cycle inhibitors, mitotic inhibitors, topoisomerase inhibitors, antihormonal drugs, angiogenesis inhibitors, cytotoxic agents.

[0258] The other active agents used in combination with the present invention can be administered simultaneously, separately, or sequentially with the compounds of the present invention via the same or different administration routes. The other active agents can be co-administered with the compounds of the present invention in a single pharmaceutical composition, or administered separately from the compounds of the present invention in different discrete units, such as combination products, preferably in the form of a kit. When administered separately, they can be administered simultaneously or successively, and the successive administrations can be close or distant in time. They can be prepared and / or formulated by the same or different manufacturers. Moreover, the compounds of the present invention and the other active agents can be (i) before sending the combination product to the physician (e.g., in the case of a kit containing the compound of the present invention and another drug); (ii) by the physician himself / herself (or under the guidance of the physician) immediately before administration; (iii) by the patient himself / herself, such as by adding them together to the combination therapy during the sequential administration of the compound of the present invention and the other active agents.

[0259] The compounds of the present invention can also be combined with anti-tumor therapies, which include, but are not limited to, surgery, radiotherapy, transplantation (such as stem cell transplantation, bone marrow transplantation), tumor immunotherapy, and chemotherapy, etc.

[0260] Therefore, on the other hand, the present invention also provides a kit, which contains two or more separate pharmaceutical compositions, at least one of which contains the compound of the present invention or its pharmaceutically acceptable salt or solvate, and a device for separately containing the compositions, such as a container, dispensing bottle, or discrete foil packaging, such as a blister pack for packaging tablets, capsules, etc., and also includes instructions for use. The kit of the present invention is particularly suitable for administering different dosage forms, such as oral dosage forms and parenteral dosage forms, or for administering different compositions at different dosing intervals.

[0261] For the technical solutions of the pharmaceutical compositions, drug combinations, or kits of the present invention described above, the abnormal cell growth involved therein, or diseases mediated by RAS, especially KRAS mutations, preferably KRAS G12C, KRAS G12D, KRAS G12V, KRAS G12A, KRAS G12R, KRAS G12S, or KRAS G13D, or KRAS Q61H mutations, and KRAS amplification are defined as above for the methods and uses of the present invention.

[0262] For the compounds, pharmaceutical compositions, methods, uses, pharmaceutical combinations and kits of the present invention described above, the compounds of the examples herein are preferred.

[0263] Preparation Methods of Compounds of the Invention

[0264] On the other hand, the present invention also provides a method for preparing the compounds defined by the present invention.

[0265] The compounds of the present invention can be prepared by a variety of methods, including the general methods given below, the methods disclosed in the examples or methods similar thereto.

[0266] Standard synthetic methods and operations for the preparation of organic compounds and for the transformation and manipulation of functional groups are known in the art and can be found in standard textbooks, such as Smith M.B., “March’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure”, 7th Edition, Wiley, 2013). For each reaction step of the general synthetic schemes, the appropriate reaction conditions are known to those skilled in the art or can be determined routinely. The method steps for synthesizing the compounds of the present invention can be carried out under reaction conditions known per se (including those specifically mentioned), in the absence or usually in the presence of a solvent or diluent (including, for example, a solvent or diluent that is inert to the reagents used and that dissolves the reagents used), in the absence or presence of a catalyst, condensing agent or neutralizing agent (such as an ion exchanger, such as a cation exchanger, such as H + form), depending on the nature of the reaction and / or the reactants, at reduced, normal or elevated temperatures (such as from about -100 °C to about 190 °C, including, for example, from about -78 °C to about 150 °C, such as from about 0 °C to about 125 °C, room temperature, -20 to 40 °C or reflux temperature), at atmospheric pressure or in a closed vessel, when appropriate under pressure, and / or in an inert atmosphere such as an argon or nitrogen atmosphere.

[0267] Unless otherwise specified, the starting materials and reagents used in the preparation of the compounds are commercially available, or compounds known in the literature, or can be prepared by those skilled in the art by the methods given below, methods similar to those given below or standard methods known in the art. Unless otherwise specified in the method description, the solvents applicable are those conventional solvents well known to those skilled in the art and suitable for the specific reaction type involved, such as water, esters, ethers, liquid aromatic hydrocarbons, alcohols, nitriles, halogenated hydrocarbons, amides, bases, carboxylic anhydrides, cyclic, straight-chain or branched-chain hydrocarbons, or mixtures of these solvents. Such solvent mixtures can also be used for work-up, for example, work-up by chromatography or partitioning.

[0268] If desired, the starting materials and intermediates in the synthetic reaction sequence can be separated and purified by conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, etc. If the intermediate and the final product are obtained in solid form, purification can also be carried out by recrystallization or aging. The materials can be characterized by conventional methods including physical constants and spectroscopic data. The reaction mixture is worked up in a conventional manner, for example by mixing with water, separating the phases, and purifying the crude product by chromatography when appropriate.

[0269] One skilled in the art can recognize the presence of stereocenters in the compounds of the present invention. At all stages of the reaction, mixtures of the isomers formed can be separated into the individual isomers, such as diastereoisomers or enantiomers, or into any desired mixture of isomers, such as a racemate or a mixture of diastereoisomers, see, for example, “Stereochemistry of Organic Compounds” by E.L. Eliel, S.H. Wilen and L.N. Mander (Wiley-Interscience, 1994).

[0270] In cases where mixtures of stereoisomers are generated during the preparation of the compounds of the present invention, the individual stereoisomers of the compounds of the present invention can be obtained by resolution, for example, by starting with the compounds of the present invention obtained as a mixture of stereoisomers and using well-known methods, such as forming diastereomeric pairs, salifying with an optically active acid, followed by fractional crystallization and regeneration of the free base, or by chiral preparative chromatography; alternatively, optically pure or enantiomerically enriched synthetic intermediates can be obtained using starting materials or intermediates with established stereochemistry, or any known chiral resolution method, and then can be used as such in subsequent steps at various stages of the above synthesis.

[0271] In certain specific cases, it may be necessary to protect specific reactive groups with suitable protecting groups to avoid interference with the reactions of other reactive groups. Suitable protecting groups and methods for protection and deprotection using such suitable protecting groups are well known to those skilled in the art; examples thereof can be found in T. Greene and P. Wuts, Protective Groups in Organic Synthesis (3rd Edition), John Wiley & Sons, NY (1999).

[0272] The following illustrates only by way of example the general synthetic schemes for synthesizing the compounds of the present invention. Other routes as well as other reactants and intermediates known to those of ordinary skill in the art can also be used to obtain the compounds of the present invention.

[0273] For the sake of clarity, in the exemplary synthesis schemes described below, unless otherwise specified, R1, R2, R3, R4, R5, R6, R7, X, Y, M, W, and t appearing in the structural formula of each intermediate compound are as defined above for the compounds of the present invention, wherein PG represents a suitable protecting group that can be determined by a person skilled in the art based on knowledge of organic chemistry.

[0274] Synthesis Scheme A

[0275] The synthesis of the compounds of the present invention can be prepared according to the following exemplary schemes or appropriate variations thereof.

[0276]

[0277] Compound 1 is commercially available or can be obtained by the methods used in the examples herein or by methods analogous thereto. In step A, compound 1 is subjected to an aromatic nucleophilic substitution reaction to obtain compound 2. Typical aromatic nucleophilic substitution conditions are well known in the art, such as DIEA / THF, etc. In step B, compound 2 is fluorinated by a halogen exchange reaction under conditions such as KF / DMSO to obtain compound 3. The latter is introduced into a naphthalene compound by a metal-catalyzed coupling reaction in step C to obtain compound 4. In step D, compound 4 is subjected to an aromatic nucleophilic substitution reaction to obtain compound 5. In step E, compound 5 is freed from any protecting groups that may be present to obtain a compound of formula I.

[0278] Typical metal-catalyzed coupling reactions include, but are not limited to, the Suzuki reaction. Typical reaction conditions are well known to those skilled in the art, and those skilled in the art understand a variety of conditions for promoting such cross-coupling reactions, such as Pd(dtbpf)Cl2 / K3PO4 / dioxane / water, or Pd(OAc)2 / rac-BIDIME / K2CO3 / toluene, or Ruphos PdG4 / K3PO4 / dioxane / water; suitable palladium catalysts also include XantPhos Pd G2, APd G3, bis(triphenylphosphine)palladium(II) chloride, Pd(dppf)Cl2, Pd2(dba)3, tetrakis(triphenyl)phosphine palladium and palladium(II) acetate, etc.; if necessary, suitable ligands may include tricyclohexylphosphine and tri-tert-butylphosphine, etc.; suitable bases also include potassium fluoride, cesium carbonate, sodium carbonate, potassium tert-butoxide and potassium phosphate monohydrate, DIPEA, etc.

[0279] It should be noted that the removal of the protecting group in step E can be adjusted according to the protecting group carried by the molecule, and can be a one-step reaction or a multi-step reaction. For example, when the protecting group PG carried by the compound is TIPS, it can be removed by reagents such as CsF.

[0280] Synthesis Scheme B

[0281] The synthesis of the compounds of the present invention can also be prepared according to the following exemplary schemes or suitable variants thereof.

[0282]

[0283] Compound 6 is commercially available or can be obtained according to the methods used in the examples herein or similar methods. In step A, compound 7 can be obtained according to the method described in synthetic scheme A. Compound 7 is then successively subjected to a metal-catalyzed coupling reaction in step B, oxidation of methyl sulfide to sulfoxide (t = 1) or sulfone (t = 2) in step C, aromatic nucleophilic substitution reaction in step D, and a possible deprotection reaction of protecting groups in step E to obtain the compound of general formula I. The typical conditions for the metal-catalyzed coupling reaction, nucleophilic substitution reaction, and deprotection reaction of protecting groups involved in this scheme are well-known in the art and can be carried out analogously with reference to the relevant reaction conditions described in synthetic scheme A.

[0284] Synthesis Examples

[0285] The present invention will be further described below in conjunction with examples. It should be noted that the following examples are exemplary and should not be regarded as limiting the protection scope of the present invention.

[0286] In the description of the embodiments and subsequent specific examples herein, the following abbreviations are used:

[0287] ACN (acetonitrile); Boc (tert - butoxycarbonyl); CDCl3 (chloroform - d); DCM (dichloromethane); DIEA or DIPEA (N,N - diisopropylethylamine); DMF (N,N - dimethylformamide); DMSO (dimethyl sulfoxide); DMSO - d6 (dimethyl sulfoxide - d6); EA (ethyl acetate); EDTA - K2 (potassium ethylenediaminetetraacetate); EtOH (ethanol); FCC (flash column chromatography); g (gram); h (hour); HCl (hydrogen chloride); HCl - MeOH or HCl / MeOH (hydrogen chloride in methanol); HLM (human liver microsomes); H2O (water); H2SO4 (sulfuric acid); IV (intravenous administration); K2CO3 (potassium carbonate); LCMS (liquid chromatography - mass spectrometry); LC - MS / MS (liquid chromatography - tandem mass spectrometry); MeOH (methanol); Methanol - d4 (methanol - d4); mg (milligram); MHz (megahertz); min (minute); mL (milliliter); mmol (millimole); MOM (methoxymethyl ether); MTBE (methyl tert - butyl ether); m / z (mass - to - charge ratio); N2 (nitrogen); NaCl (sodium chloride); NaH (sodium hydride); NaHCO3 (sodium bicarbonate); Na2SO3 (sodium sulfite); Na2SO4 (sodium sulfate); NCS (N - chlorosuccinimide); NH4Cl (ammonium chloride); NMR (nuclear magnetic resonance); PdCl2(dtbpf) or Pd(dtbpf)Cl2 (1,1'-bis(di - tert - butylphosphino)ferrocene dichloropalladium); PdCl2(dppf) or Pd(dppf)Cl2 (1,1'-bis(diphenylphosphino)ferrocene dichloropalladium); Pd(OAc) (palladium(II) acetate); Pd(PPh3)4 (tetrakis(triphenylphosphine)palladium(0)); PE (petroleum ether); PO (oral administration); POCl3 (phosphorus oxychloride); r.t. (room temperature); SiO2 (silica gel); TEA (triethylamine); TFA (trifluoroacetic acid); THF (tetrahydrofuran); TIPS (triisopropylsilyl); TLC (thin - layer chromatography); TsOH (p - toluenesulfonic acid); TsOH·H2O (p - toluenesulfonic acid monohydrate); μL (microliter); μM (micromolar concentration); μmol (micromole).

[0288] In the following examples, the names and structures of the synthesized target compounds are given. Any deviation between the name and the structure is not intentional, and in such cases, the structure is decisive.

[0289] The experimental methods without specific conditions noted in the following examples are generally carried out according to the conventional conditions of such reactions or according to the conditions recommended by the manufacturers. Unless otherwise stated, percentages and parts are by weight. Unless otherwise stated, the ratio of liquids is by volume.

[0290] The experimental materials and reagents used in the following examples can be obtained from commercial channels, prepared according to the methods of the prior art, or prepared according to methods similar to those disclosed in this application, unless otherwise specified.

[0291] In the following examples, 1 1H-NMR spectra were recorded on a Bruker (400 MHz), and chemical shifts were expressed as δ (ppm) relative to the deuterated solvent peak (CDCl3: δ = 7.26 ppm; CD3OD: δ = 3.31 ppm; DMSO-d6: δ = 2.50 ppm); liquid chromatography-mass spectrometry was recorded using an Agilent 1260 liquid chromatography + Agilent G6125B mass spectrometry LCMS liquid chromatography-mass spectrometry instrument. The gas chromatography-mass spectrometry was detected using a Shimadzu GCMS-QP2010SE.

[0292] Intermediate A

[0293]

[0294] 7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one

[0295]

[0296] Step A: 4,6-dichloro-5-fluoronicotinoyl chloride

[0297] With stirring at room temperature, thionyl chloride (2.25 mL, 31 mmol) was slowly added to a solution of 4,6-dichloro-5-fluoronicotinic acid (5.0 g, 23.4 mmol) in DCM (100 mL), and then DMF (175 mg, 2.4 mmol) was added. The resulting reaction solution was stirred at 50 °C for 2 h. After monitoring the reaction to completion by TLC, it was concentrated, and after adding a small amount of toluene for co-evaporation, yellow solid 4,6-dichloro-5-fluoronicotinoyl chloride (4.5 g, yield 83%) was obtained and directly used for the subsequent reaction.

[0298] Step B: (4,6-dichloro-5-fluoronicotinoyl)carbamimidothioate

[0299] With stirring at 0 °C, a mixed solution of 4,6-dichloro-5-fluoronicotinoyl chloride (4.5 g, 19.8 mmol) and ethylene glycol dimethyl ether (20 ml) was slowly added dropwise to a mixed solution of 2-methylisothiourea sulfate (15 g, 49.5 mmol) and 1 M aqueous NaOH solution (70 ml), and the temperature was maintained while stirring for 1 h. The precipitated solid was filtered and dried to obtain the product (4,6-dichloro-5-fluoronicotinoyl)carbamimidothioate (5.0 g, yield 90%). LCMS (m / z): 282.1 (M+H).

[0300] Step C: 7-Chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one

[0301] Dissolve (4,6-dichloro-5-fluoronicotinoyl)carbamimidothioate (5.0 g, 17.8 mmol) in DMF (40 mL), heat to 120 °C and stir the reaction for 3 h. After monitoring the completion of the reaction by LCMS, cool to room temperature and add water (200 mL). Filter and dry the precipitated solid to obtain the product (4,6-dichloro-5-fluoronicotinoyl)carbamimidothioate (3.6 g, yield 82%). LCMS (m / z): 245.6 (M+H).

[0302] Intermediate B

[0303]

[0304] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one

[0305]

[0306] Step A: 2,6-Dichloro-3-fluoropyridin-4-amine

[0307] At room temperature, add Selectfluor (68 g, 180 mmol) to a solution of 2,6-dichloropyridin-4-amine (25 g, 154 mmol) in methanol / water (V / V = 5:1, 300 mL). Stir the resulting mixture at 50 °C for 48 h, concentrate under reduced pressure, dilute with ethyl acetate, wash successively with water and saturated brine, and dry over anhydrous sodium sulfate. Filter and concentrate, and purify the obtained crude product by FCC (SiO2, EA / PE = 0-10%) to obtain white solid 2,6-dichloro-3-fluoropyridin-4-amine (10 g). LCMS (m / z): 180.9 (M+H).

[0308] Step B: tert-Butyl (2,6-dichloro-3-fluoropyridin-4-yl)carbamate

[0309] Under stirring at room temperature, 4-dimethylaminopyridine (307 mg, 2.75 mmol) and di-tert-butyl dicarbonate (30 g, 138 mmol) were added to a solution of 2,6-dichloro-3-fluoropyridin-4-amine (10 g, 55 mmol) in tetrahydrofuran (100 mL). The resulting mixture was heated to 60 °C and stirred for 16 h. The reaction was monitored by TLC and completed. The mixture was concentrated to obtain a crude product, which was slurried with methanol to give a white solid, tert-butyl ((2,6-dichloro-3-fluoropyridin-4-yl)carbamoyl)carbamate (16 g). LCMS (m / z): 381.2 (M+H).

[0310] Step C: tert-butyl 4-((tert-butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinate

[0311] Under a dry ice-ethanol bath, LDA (2.0 M, 63 mL, 126 mmol) was slowly added to a solution of tert-butyl ((2,6-dichloro-3-fluoropyridin-4-yl)carbamoyl)carbamate (16 g, 42 mmol) in THF (200 mL). The resulting mixture was stirred at the same temperature for 1 h. The reaction was monitored by TLC and completed. An appropriate amount of acetic acid was added to quench the reaction, and the mixture was diluted with EA, washed with water, and dried over anhydrous sodium sulfate. After filtration and concentration, the obtained crude product was purified by FCC (SiO2, EA / PE = 0-20%) to give tert-butyl 4-((tert-butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinate (13 g).

[0312] Step D: 4-amino-2,6-dichloro-5-fluoronicotinic acid hydrochloride

[0313] Concentrated hydrochloric acid (30 ml) was added to a solution of tert-butyl 4-((tert-butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinate (13 g, 34 mmol) in dioxane (90 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was monitored by LCMS and completed, it was concentrated to give 4-amino-2,6-dichloro-5-fluoronicotinic acid hydrochloride (8 g). LCMS (m / z): 224.9 (M+H).

[0314] Step E: 5,7-dichloro-8-fluoro-2-mercaptopyrido[4,3-d]pyrimidin-4(3H)-one

[0315] A mixed solution of 4-amino-2,6-dichloro-5-fluoronicotinic acid (8 g, 30.8 mmol) and thionyl chloride (200 mL) was stirred at 50 °C for 3 h. Then it was concentrated, and the residue was dissolved in acetone (50 mL) to obtain Solution 1. At room temperature, a mixed solution of ammonium thiocyanate (7 g, 92 mmol) and acetone solution (160 mL) was dropped into Solution 1, and the resulting reaction solution was continuously stirred at room temperature for 1 h. After the reaction was monitored by LCMS to be completed, the reaction solution was poured into water, filtered, and the filter cake was dried to obtain 5,7-dichloro-8-fluoro-2-mercaptopyrido[4,3-d]pyrimidin-4(3H)-one (5 g). LCMS (m / z): 265.9 (M+H).

[0316] Step F: 5,7-Dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one

[0317] At room temperature, a mixed solution of 5,7-dichloro-8-fluoro-2-mercaptopyrido[4,3-d]pyrimidin-4(3H)-one (5 g, 18.8 mmol), methanol (380 mL), aqueous sodium hydroxide solution (0.1 M, 380 mL, 380 mmol) and methyl iodide (5.3 g, 380 mmol) was stirred for 2 h. After the reaction was monitored by LCMS to be completed, the reaction solution was poured into 1000 ml of water and acidified to pH~6 with concentrated hydrochloric acid. The solution was filtered, and the filter cake was dried to obtain the product 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (4 g). LCMS (m / z): 279.9 (M+H).

[0318] Step G: 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one

[0319] A mixture of 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (400 mg, 1.4 mmol), sodium methoxide (0.38 g, 7.5 mmol), DMA (10 mL) and methanol (2 mL) was stirred at 50 °C for 16 h. After the reaction was monitored by LCMS to be completed, it was diluted with water, adjusted to pH~3 with concentrated hydrochloric acid, filtered, the filter cake was collected and dried to obtain the product 7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (250 mg). LCMS (m / z): 276.0 (M+H).

[0320] Intermediate C

[0321]

[0322] 7-Bromo-2,4-dichloro-6,8-difluoroquinazoline

[0323]

[0324] Step A: Methyl 4-bromo-3,5-difluoro-2-(3-(2,2,2-trichloroacetyl)ureido)benzoate

[0325] At room temperature, methyl 2-amino-4-bromo-3,5-difluorobenzoate (10 g, 37.6 mmol) and THF (100 mL) were added to a round-bottom flask equipped with a magnetic stirrer. 2,2,2-Trichloroacetyl isocyanate (8.5 g, 45.1 mmol) was added dropwise to the system under stirring at room temperature. The resulting mixture was stirred at room temperature for 2 hours and concentrated under reduced pressure to obtain a brown solid, methyl 4-bromo-3,5-difluoro-2-(3-(2,2,2-trichloroacetyl)ureido)benzoate (crude product), which was directly used in the next step. LCMS (ESI, m / z): 452.8 (M+H).

[0326] Step B: 7-Bromo-6,8-difluoroquinazoline-2,4-diol

[0327] At room temperature, methyl 4-bromo-3,5-difluoro-2-(3-(2,2,2-trichloroacetyl)ureido)benzoate obtained in the previous step was added to a round-bottom flask equipped with a magnetic stirrer, and then NH3 (100 mL, 7M MeOH solution) was added. The resulting mixture was stirred at room temperature for 2 hours, and the reaction was monitored by LCMS until completion. The mixture was concentrated under reduced pressure, and the resulting solid was slurried with methyl tert-butyl ether and filtered to obtain a pale yellow solid, 7-bromo-6,8-difluoroquinazoline-2,4-diol (14 g, crude product), which was directly used in the next step without further purification. LCMS (ESI, m / z): 277.0 (M+H).

[0328] Step C: 7-Bromo-2,4-dichloro-6,8-difluoroquinazoline

[0329] 7-Bromo-6,8-difluoroquinazoline-2,4-diol (14 g, crude product) and POCl3 (112 mL) were added to a round-bottom flask equipped with a magnetic stirrer. DIEA (28 mL) was added dropwise to the system under stirring at room temperature. After the addition was complete, the system was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure to about 30 mL, poured into water (600 mL), and the precipitate was filtered, collected, dried, and a yellow solid, 7-bromo-2,4-dichloro-6,8-difluoroquinazoline (11 g, crude product), was obtained, which was directly used in the subsequent reaction. LCMS (ESI, m / z): 312.9 (M+H).

[0330] Following the above method and appropriate variations known in the art, the following intermediates were prepared:

[0331]

[0332]

[0333] Intermediate A-1

[0334]

[0335] 6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0336]

[0337] Step A: 4,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine

[0338] Under N2, DIEA (1.0 mL, 6.1 mmol) was added to a solution of 7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-ol (1.0 g, 4.1 mmol) in phosphorus oxychloride (10 mL). The resulting reaction mixture was stirred at 90 °C for 1 h. After monitoring the reaction by LCMS until completion, the reaction mixture was concentrated to dryness. The obtained crude product was purified by FCC (SiO2, EA / PE = 0 - 10%) to give the yellow solid product 4,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine (810 mg, yield 75%). LCMS (m / z): 263.9 (M+H).

[0339] Step B: 6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0340] Under an ice bath, DIEA (792 mg, 6.2 mmol) and 4,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine (810 mg, 3.1 mmol) were successively added to a solution of 1-oxa-6-azaspiro[3.5]nonane·oxalate (660 mg, 3.1 mmol) in DMF (10 mL). The resulting reaction mixture was stirred at 0 °C for 0.5 h. After monitoring the reaction by LCMS until completion, ethyl acetate (80 mL) was added to the reaction mixture. The resulting reaction mixture was washed with water, dried, filtered and concentrated to dryness. The obtained crude product was purified by FCC (SiO2, EA / PE = 0 - 20%) to give the yellow solid product 6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (980 mg, yield 90%). LCMS (m / z): 355.0 (M+H).

[0341] Intermediate A-I-A and Intermediate A-I-B

[0342]

[0343] (R)-6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane and (S)-6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0344]

[0345] Step A: Benzyl (S)-1-oxa-6-azaspiro[3.5]nonane-6-carboxylate and benzyl (R)-1-oxa-6-azaspiro[3.5]nonane-6-carboxylate

[0346] Under nitrogen at 50 °C, a solution of sodium tert-butoxide (428 mL, 428 mmol, 1 N THF solution) was added to a solution of trimethylsulfoxonium iodide (94 g, 428 mmol) in tert-butanol (200 mL). After the resulting reaction mixture was stirred at 50 °C for 1.5 h, 3-oxopiperidine-1-carboxylic acid benzyl ester (25 g, 107 mmol) was added to the above reaction mixture, and the resulting reaction mixture was further reacted at 50 °C under nitrogen for 16 h. After the reaction was monitored by LCMS and completed, it was cooled to room temperature, and saturated NH4Cl solution (200 mL) was added to the reaction mixture. It was extracted with EtOAc, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness, and the obtained crude product was purified by FCC (SiO2, EA / PE = 0-20%) to obtain a colorless transparent liquid product, benzyl 1-oxa-6-azaspiro[3.5]nonane-6-carboxylate (10 g, yield 36%). LCMS (m / z): 262.1 (M+H). 1 H NMR (400 MHz, Chloroform-d) δ 7.41–7.27 (m, 5H), 5.14 (s, 2H), 4.63–4.42 (m, 2H), 3.82 (d, J = 13.0 Hz, 1H), 3.65–3.11 (m, 3H), 2.49–2.25 (m, 2H), 1.96–1.64 (m, 3H), 1.57–1.38 (m, 1H).

[0347] The above colorless transparent liquid product was resolved by SFC (Waters SFC 150, REGIS (S,S) WHELK-O1 (250 * 40mm 10μm), Supercritical CO2 / MeOH (0.1% DEA in MeOH) = 80 / 20), and the first eluted isomer was (R)-benzyl 1-oxa-6-azaspiro[3.5]nonane-6-carboxylate (4.1 g, yield 15%, with a relatively small relative retention time). 1 1H NMR (400 MHz, Chloroform-d) δ 7.43–7.26 (m, 5H), 5.14 (s, 2H), 4.65–4.43 (m, 2H), 3.82 (d, J = 13.0 Hz, 1H), 3.63–3.13 (m, 3H), 2.49–2.22 (m, 2H), 1.96–1.63 (m, 4H), 1.56–1.33 (m, 1H). LCMS (m / z): 262.1 (M+H). SFC analysis method A-1-2, Rt = 2.584 min. The subsequently eluted isomer was (S)-benzyl 1-oxa-6-azaspiro[3.5]nonane-6-carboxylate (4.5 g, yield 16%, with a relatively large relative retention time). 1 1H NMR (400 MHz, Chloroform-d) δ 7.43–7.27 (m, 5H), 5.14 (s, 2H), 4.64–4.40 (m, 2H), 3.82 (d, J = 13.0 Hz, 1H), 3.62–3.05 (m, 3H), 2.50–2.18 (m, 2H), 2.01–1.65 (m, 3H), 1.54–1.37 (m, 1H). LCMS (m / z): 262.1 (M+H). SFC analysis method A-1-2, Rt = 4.563 min.

[0348] SFC analysis method A-1-2: Waters UPCC (CA-352), analytical column: Daicel IG, 100 * 3mm * 3μm; mobile phase A: CO2, mobile phase B: MeOH (0.1% DEA); flow rate: 1.5 mL / min; column temperature: 35 °C; back pressure: 1800 psi; gradient: 0 - 8.0 min A / B = 80 / 20.

[0349] Step B: (R)-1-oxa-6-azaspiro[3.5]nonane oxalate · oxalate and (S)-1-oxa-6-azaspiro[3.5]nonane oxalate · oxalate

[0350] Dissolve benzyl (R)-1-oxa-6-azaspiro[3.5]nonane-6-carboxylate (4.1 g, 15.6 mmol) in methanol (50 mL). Under N2 atmosphere, add Pd / C (1.6 g, 1.56 mmol, 10% w / w), displace with H2 (15 psi) three times, and stir at room temperature for 8 h. Monitor the disappearance of the starting material by LCMS. Filter the reaction mixture through diatomaceous earth, add oxalic acid (1 g) to form a salt, and concentrate to obtain the white solid (R)-1-oxa-6-azaspiro[3.5]nonane oxalate·oxalate (3.0 g). 1 H NMR (400 MHz, D2O) δ 4.71–4.51 (m, 2H), 3.65–3.57 (m, 1H), 3.29–3.19 (m, 2H), 3.08–2.95 (m, 1H), 2.65–2.41 (m, 2H), 2.26–2.11 (m, 1H), 1.96–1.71 (m, 3H).

[0351] Dissolve benzyl (S)-1-oxa-6-azaspiro[3.5]nonane-6-carboxylate (4.5 g, 17.2 mmol) in methanol (50 mL). Under N2 atmosphere, add Pd / C (1.8 g, 1.7 mmol, 10%), displace with H2 (15 psi) three times, and stir at room temperature for 8 h. Monitor the disappearance of the starting material by LCMS. Filter the reaction mixture through diatomaceous earth, add oxalic acid (1 g) to form a salt, and concentrate to obtain the white solid (S)-1-oxa-6-azaspiro[3.5]nonane oxalate·oxalate (3.5 g). 1 H NMR (400 MHz, D2O) δ 4.72–4.52 (m, 2H), 3.67–3.55 (m, 1H), 3.30–3.18 (m, 2H), 3.08–2.95 (m, 1H), 2.64–2.44 (m, 2H), 2.25–2.12 (m, 1H), 1.95–1.74 (m, 3H).

[0352] Step C: (R)-6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane and (S)-6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0353] The experimental procedure was carried out according to the protocol described for Intermediate A-I, using (R)-1-oxa-6-azaspiro[3.5]nonane oxalate·oxalate and (S)-1-oxa-6-azaspiro[3.5]nonane oxalate·oxalate.

[0354] Intermediate A-II

[0355]

[0356] 7-Chloro-8-fluoro-2-(methylthio)-4-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-1,3,6-triazaindene

[0357]

[0358] Step A: 2-Oxa-6-azabicyclo[5.1.0]octane p-toluenesulfonate

[0359] At room temperature, dissolve benzyl 2-oxa-6-azabicyclo[5.1.0]octane-6-carboxylate (750 mg, 3.03 mmol) and Pd / C (718 mg, 10% w / w, 0.61 mmol) in methanol (15 mL), displace with H2 (15 psi) twice, and stir at room temperature for 2 h. After the reaction is completed as detected by LCMS, the reaction solution is filtered through diatomaceous earth. After the organic phase is completely concentrated, white solid 2-oxa-6-azabicyclo[5.1.0]octane p-toluenesulfonate (750 mg, yield 86%) is obtained. LCMS (m / z): 114.1 (M+H).

[0360] Step B: 7-Chloro-8-fluoro-2-(methylthio)-4-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-1,3,6-triazaindene

[0361] At room temperature, add DIEA (1.25 mL, 7.57 mmol) to a solution of 2-oxa-6-azabicyclo[5.1.0]octane p-toluenesulfonate (750 mg, 2.63 mmol) and 4,7-dichloro-8-fluoro-2-(methylthio)-1,3,6-triazaindene (500 mg, 1.89 mmol) in THF (15 mL). Stir at room temperature for 1 h. After the reaction is completed as monitored by LCMS, the reaction solution is concentrated, 5 mL of acetonitrile is added to dissolve the oil, and it is poured into H2O (100 mL). A brownish-yellow solid precipitates. Filter, wash the filter cake with H2O (100 mL), and dry to obtain brown solid 7-chloro-8-fluoro-2-(methylthio)-4-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-1,3,6-triazaindene (640 mg, yield 99%). LCMS (m / z): 341.0 (M+H).

[0362] Intermediate A-II-A and A-II-B

[0363]

[0364] 4-((1S,7R)-2-Oxo-6-aza-bicyclo[5.1.0]oct-6-yl)-7-chloro-8-fluoro-2-(methylthio)-1,3,6-triazaindene and 4-((1R,7S)-2-oxo-6-aza-bicyclo[5.1.0]oct-6-yl)-7-chloro-8-fluoro-2-(methylthio)-1,3,6-triazaindene

[0365]

[0366] Step A: Benzyl (1R,7S)-2-oxa-6-aza-bicyclo[5.1.0]octane-6-carboxylate and benzyl (1S,7R)-2-oxa-6-aza-bicyclo[5.1.0]octane-6-carboxylate

[0367] At 0 °C, ZnEt2 (62.27 mL, 62.3 mmol) was added to a solution of benzyl 4,5,6,7-tetrahydro-1,4-oxepane-4-carboxylate (5.8 g, 24.9 mmol) in DCM (80 mL). The resulting reaction mixture was warmed to room temperature and stirred for 0.5 h. A solution of diiodomethane (8.03 mL, 174 mmol) in DCM (30 mL) was added to the above reaction mixture, and the reaction was continued to stir at room temperature for 3 h. After the reaction was monitored by LCMS to completion, the reaction mixture was poured into a semi-saturated aqueous solution of NH4Cl (200 mL), and extracted with DCM (50 mL × 3). The organic layer was collected, washed with saturated brine (20 mL), dried over anhydrous Na2SO4, and concentrated. The resulting crude product was purified by FCC (SiO2, PE / EA = 0 - 40%) to obtain a pale yellow oily product, benzyl 2-oxa-6-aza-bicyclo[5.1.0]octane-6-carboxylate (5 g, yield 81.3%), LCMS (m / z): 248.0 (M+H).

[0368] The above pale yellow oily product was resolved by SFC (Waters SFC 150, REGIS (S,S) WHELK-O1 (250*40mm 10μm), CO2 / MeOH (0.1% DEA-MeOH) = 90 / 10). The first eluted isomer was benzyl (1R,7S)-2-oxa-6-aza-bicyclo[5.1.0]octane-6-carboxylate (2.2 g, yield 44%, with a relatively small retention time). SFC analysis method A-1I-2, Rt = 1.505 min. The subsequently eluted isomer was benzyl (1S,7R)-2-oxa-6-aza-bicyclo[5.1.0]octane-6-carboxylate (2.1 g, yield 42%, with a relatively large retention time). SFC analysis method A-1I-2, Rt = 2.261 min.

[0369] SFC Analytical Method A-1I-2: Waters UPCC (CA-415), Analytical Column: Daicel AD, 100 * 3 mm * 3 μm; Mobile Phase A: CO2, Mobile Phase B: MeOH (0.1% DEA); Flow Rate: 1.5 mL / min; Column Temperature: 35 °C; Back Pressure: 1800 psi; Gradient: 0 - 4.0 min A / B = 90 / 10.

[0370] Step B: (1R,7S)-2-Oxa-6-azabicyclo[5.1.0]octane p-toluenesulfonate and (1S,7R)-2-Oxa-6-azabicyclo[5.1.0]octane p-toluenesulfonate

[0371] The experimental procedure was carried out according to the protocol described for Intermediate A-II, using (1R,7S)-2-Oxa-6-azabicyclo[5.1.0]octane-6-carboxylic acid benzyl ester and (1S,7R)-2-Oxa-6-azabicyclo[5.1.0]octane-6-carboxylic acid benzyl ester.

[0372] Step C: 4-((1R,7S)-2-Oxa-6-azabicyclo[5.1.0]oct-6-yl)-7-chloro-8-fluoro-2-(methylthio)-1,3,6-triazaindene and 4-((1S,7R)-2-Oxa-6-azabicyclo[5.1.0]oct-6-yl)-7-chloro-8-fluoro-2-(methylthio)-1,3,6-triazaindene

[0373] The experimental procedure was carried out according to the protocol described for Intermediate A-II, using (1R,7S)-2-Oxa-6-azabicyclo[5.1.0]octane p-toluenesulfonate and (1S,7R)-2-Oxa-6-azabicyclo[5.1.0]octane p-toluenesulfonate.

[0374] Intermediate B-1

[0375]

[0376] 6-(7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0377]

[0378] The synthesis of Intermediate B-I was carried out by referring to the synthesis of Intermediate A-I, using 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-ol in Step A. LCMS (m / z): 385.1 (M + H).

[0379] Intermediate C-1

[0380]

[0381] 4-((S)-1-Oxa-6-aza-6-spiro[3.5]nonyl)-7-bromo-2,6,8-trifluoroquinazoline

[0382]

[0383] Step A: 4-((S)-1-Oxa-6-aza-6-spiro[3.5]nonyl)-7-bromo-2-chloro-6,8-difluoroquinazoline

[0384] Under an ice bath, DIEA (617 mg, 4.8 mmol) and 7-bromo-2,4-dichloro-6,8-difluoroquinazoline (500 mg, 1.6 mmol) were successively added to a solution of (S)-1-oxa-6-azaspiro[3.5]nonyl oxalate oxalate (380 mg, 1.7 mmol) in DMF (10 mL). The resulting reaction solution was stirred at 0 °C for 1 h. After monitoring the reaction by LCMS until completion, ethyl acetate (80 mL) was added to the reaction solution. The resulting reaction solution was washed with water, dried, filtered and concentrated to dryness. The obtained crude product was purified by FCC (SiO2, EA / PE = 0 - 25%) to obtain a yellow solid product 4-((S)-1-oxa-6-aza-6-spiro[3.5]nonyl)-7-bromo-2-chloro-6,8-difluoroquinazoline (550 mg, yield 85%). LCMS (m / z): 403.9 (M+H).

[0385] Step B: 4-((S)-1-Oxa-6-aza-6-spiro[3.5]nonyl)-7-bromo-2,6,8-trifluoroquinazoline

[0386] At room temperature, KF (789 mg, 13 mmol) was added to a solution of 4-((S)-1-oxa-6-aza-6-spiro[3.5]nonyl)-7-bromo-2-chloro-6,8-difluoroquinazoline (550 mg, 1.3 mmol) in DMSO (10 mL). The resulting reaction solution was stirred at 110 °C for 12 h. After monitoring the reaction by LCMS until completion, ethyl acetate (50 mL) was added to the reaction solution. The resulting reaction solution was washed with water, dried, filtered and concentrated to dryness. The obtained crude product was purified by FCC (SiO2, EA / PE = 0 - 20%) to obtain a yellow solid product 4-((S)-1-oxa-6-aza-6-spiro[3.5]nonyl)-7-bromo-2,6,8-trifluoroquinazoline (430 mg, yield 81%). LCMS (m / z): 390.0 (M+H).

[0387] Intermediate a

[0388]

[0389] (R)-1-(tert-Butyl) 3-methyl 4-oxopiperidine-1,3-dicarboxylate

[0390]

[0391] The compound 1-(tert-butyl) 3-methyl 4-oxopiperidine-1,3-dicarboxylate 3-methyl ester (120 g) was resolved by SFC (SFC150, Waters) (separation column: DAICEL IG, 250*50 mm, 10 μm; mobile phase: CO2 / MeOH = 90 / 10; flow rate: 120 mL / min), and the first eluted isomer 1, which is compound a (52.8 g, with a relatively smaller retention time), was obtained. Chiral analysis method - a, Rt = 0.682 min. 1 1H NMR (400 MHz, Chloroform-d) δ 4.59–4.42 (m, 1H), 4.26–3.98 (m, 1H), 3.73 (s, 3H), 3.42–3.24 (m, 1H), 3.16–3.01 (m, 1H), 2.93–2.63 (m, 1H), 2.58–2.40 (m, 1H), 1.49 (s, 9H), 1.31 (s, 3H). LCMS (m / z): 216.1 (M - 56 + H). The subsequently eluted isomer 2, which is compound a-1 (52.4 g, with a relatively larger retention time), was obtained. Chiral analysis method - a, Rt = 1.035 min. 1 1H NMR (400 MHz, Chloroform-d) δ 4.60–4.41 (m, 1H), 4.24–3.94 (m, 1H), 3.73 (s, 3H), 3.42–3.24 (m, 1H), 3.17–3.00 (m, 1H), 2.93–2.64 (m, 1H), 2.56–2.40 (m, 1H), 1.49 (s, 9H), 1.31 (s, 3H).

[0392] Chiral analysis method - a: (Waters UPCC, analytical column: Daicel IG, 100*3 mm 3 μm; mobile phase A: CO2, mobile phase B: MeOH; flow rate: 1.5 mL / min; column temperature: 35 °C; back pressure: 1800 psi; gradient: 0 - 8.0 min A / B = 90 / 10).

[0393] Intermediate b

[0394]

[0395] (S,E)-(4-(Fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol

[0396]

[0397] Step A: (S,E)-4-(Fluoromethylene)-3-methylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester 3-methyl ester and (S,Z)-4-(Fluoromethylene)-3-methylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester 3-methyl ester

[0398] Dissolve (fluoromethylene)triphenylphosphonium tetrafluoroborate (10.56 g, 27.64 mmol) in anhydrous THF (50 mL), and displace the air with nitrogen three times. Under the condition of dry ice - ethanol, cool the reaction solution to -70 °C, and slowly dropwise add a solution of potassium tert-butoxide - THF (27.64 mL, 1 M, 27.64 mmol) to the reaction system. Keep stirring at this temperature for 1 h. Then, slowly dropwise add a solution of (R)-3-methyl-4-oxopiperidine-1,3-dicarboxylic acid 1-(tert-butyl)-3-methyl ester (5.0 g, 18.43 mmol) in anhydrous THF (15 mL) to the reaction system. After the addition is complete, slowly raise the temperature of the resulting mixture to room temperature and stir overnight. After monitoring the reaction by TLC until completion, slowly pour the reaction solution into water (100 mL), and extract with ethyl acetate three times. Combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, filter and concentrate to dryness to obtain the crude product. The crude product is purified by FCC (SiO2, EA / PE = 0 - 15%) to obtain a colorless oily product (S,E)-4-(fluoromethylene)-3-methylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester 3-methyl ester (1.98 g, yield 37%). LCMS (m / z): 232.1 (M - 56 + H). 1 1H NMR (400 MHz, Chloroform-d) δ 6.55 (d, J = 84.7, 1H), 4.35 (d, J = 13.2 Hz, 1H), 4.10–3.82 (m, 1H), 3.69 (s, 3H), 3.00–2.85 (m, 1H), 2.76 (d, J = 13.1 Hz, 1H), 2.71–2.60 (m, 1H), 2.31–2.08 (m, 1H), 1.46 (s, 9H), 1.29 (s, 3H); and a colorless oily product (S,Z)-4-(fluoromethylene)-3-methylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester 3-methyl ester (600 mg, yield 11%). LCMS (m / z): 232.1 (M - 56 + H). 11H NMR (400 MHz, Chloroform-d) δ 6.43 (d, J = 83.7, 1H), 3.86–3.75 (m, 1H), 3.71 (s, 3H), 3.62–3.47 (m, 1H), 3.42–3.29 (m, 2H), 2.24–2.06 (m, 2H), 1.46 (s, 9H), 1.43–1.39 (m, 3H).

[0399] Step B: Methyl (S,E)-4-(fluoromethylene)-3-methylpiperidine-3-carboxylate hydrochloride

[0400] At room temperature, 4M HCl-dioxane (10 mL) was added to (S,E)-tert-butyl 4-(fluoromethylene)-3-methylpiperidine-1,3-dicarboxylate 3-methyl ester (600 mg, 2.09 mmol), and the mixture was stirred at room temperature for 1 h. The acid solution was concentrated and removed to obtain the white solid methyl (S,E)-4-(fluoromethylene)-3-methylpiperidine-3-carboxylate hydrochloride (572 mg, yield 100%). LCMS (m / z): 188.1 (M+H).

[0401] Step C: Methyl (S,E)-4-(fluoromethylene)-1,3-dimethylpiperidine-3-carboxylate

[0402] At room temperature, methyl (S,E)-4-(fluoromethylene)-3-methylpiperidine-3-carboxylate hydrochloride (370 mg, 1.98 mmol) was dissolved in methanol (5 mL), and triethylamine was added dropwise until the pH of the reaction solution was ~10. The mixture was stirred for 10 minutes, and then glacial acetic acid was added dropwise until the pH of the reaction solution was ~4. An aqueous formaldehyde solution (481.15 mg, 5.93 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 30 min. Sodium cyanoborohydride (136.62 mg, 2.17 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 2 h. After monitoring the reaction by LCMS and completion, the solvent was removed by concentration under reduced pressure, and the residue was co-evaporated twice with anhydrous tetrahydrofuran to obtain the white solid methyl (S,E)-4-(fluoromethylene)-1,3-dimethylpiperidine-3-carboxylate (380 mg, yield 96%). LCMS (m / z): 202.1 (M+H).

[0403] Step D: (S,E)–(4-(Fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol

[0404] Under ice bath conditions, a 1 M solution of LiAlH4 in THF (2.83 mL, 107.5 mg, 2.83 mmol) was added dropwise to a solution of methyl (E)-4-(fluoromethylene)-1,3-dimethylpiperidine-3-carboxylate (380 mg, 1.89 mmol) in anhydrous THF (5 mL). The resulting mixture was stirred at room temperature for 20 min. After monitoring the reaction by LCMS until completion, the reaction solution was quenched with sodium sulfate decahydrate until no bubbles were generated. Approximately 5 g of anhydrous sodium sulfate was added to remove water. The reaction solution was filtered through diatomaceous earth, and the filter cake was washed three times with anhydrous THF. The filtrate was collected and concentrated to dryness to obtain a colorless oily product, (S,E)–(4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol (300 mg, yield 92%). LCMS (m / z): 174.1 (M+H).

[0405] Intermediate c

[0406]

[0407] ((3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methanol

[0408]

[0409] Step A: (S)-4-(Difluoromethylene)-3-methylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl)-3-methyl ester

[0410] (R)-1-(tert-Butyl)-3-methyl 4-oxopiperidine-1,3-dicarboxylate (10.0 g, 36.86 mmol) and 2-((difluoromethyl)sulfonyl)pyridine (10.68 g, 55.29 mmol) were dissolved in anhydrous DMF (100 mL), and the system was purged with nitrogen three times. The system was cooled with a dry ice-ethanol bath, and a 1 molar solution of potassium tert-butoxide in THF (66.34 mL, 66.34 mmol) was added dropwise. The resulting mixture was stirred at the same temperature for 2 h, then slowly warmed to room temperature and stirred for an additional 3 h. After monitoring the reaction by LCMS until completion, the reaction was quenched with saturated aqueous ammonium chloride (50 mL), and water (200 mL), DCM / MeOH (100 mL, v / v = 10 / 1) was used for extraction 5 times. The organic layer was washed 3 times with an aqueous LiCl solution (100 mL, 4% w / w), saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The resulting crude product was purified by FCC (SiO2, EA / PE = 0-20%) to obtain a pale yellow oily product (5.2 g, yield 46%). LCMS (m / z): 250.1 (M-56+H).

[0411] Step B: (3S,4S)-4-(Difluoromethyl)-3-methylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester

[0412] Under room temperature conditions, dissolve (S)-4-(difluoromethylene)-3-methylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester (6.20 g, 20.31 mmol) in methanol (150 mL), and displace with nitrogen three times. Add palladium on carbon (2.16 g, 10% w / w), and displace with hydrogen three times. Stir at 30 °C for 4 h under a hydrogen atmosphere of 15 Psi. After monitoring the reaction by LCMS and completion of the reaction, filter the reaction solution through diatomaceous earth, and wash the filter cake with methanol three times. Collect the filtrate, concentrate to dryness to obtain the colorless oily product (3S,4S)-4-(difluoromethyl)-3-methylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester (5.53 g, yield 89%). LCMS (m / z): 252.1 (M - 56 + H).

[0413] Step C: (3S,4S)-4-(Difluoromethyl)-3-methylpiperidine-3-carboxylic acid methyl ester hydrochloride

[0414] Under room temperature conditions, dissolve (3S,4S)-4-(difluoromethyl)-3-methylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester (5.53 g, 17.99 mmol) in 4M HCl / dioxane (60 mL), stir the resulting mixture at room temperature for 1 h, concentrate to remove the acid solution to obtain the white solid (3S,4S)-4-(difluoromethyl)-3-methylpiperidine-3-carboxylic acid methyl ester hydrochloride (4.38 g, yield 100%). LCMS (m / z): 208.1 (M + H).

[0415] Step D: (3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidine-3-carboxylic acid methyl ester

[0416] At room temperature, (3S,4S)-methyl 4-(difluoromethyl)-3-methylpiperidine-3-carboxylate hydrochloride (3.58 g, 14.69 mmol) was dissolved in methanol (40 mL), and aqueous formaldehyde solution (3.58 g, 37% w / w, 44.07 mmol) was added. The resulting mixture was stirred at room temperature for 30 min. Sodium cyanoborohydride (1.11 g, 17.63 mmol) was added, and the resulting mixture was stirred at room temperature for 1.5 h. After the reaction was monitored by LCMS and completed, the system was concentrated to dryness, ethyl acetate was added to dissolve the crude product, and the mixture was filtered through diatomaceous earth. The obtained filtrate was purified by FCC (SiO2, MeOH / DCM (containing 0.3% DIEA) = 0 - 4%) to obtain (3S,4S)-methyl 4-(difluoromethyl)-1,3-dimethylpiperidine-3-carboxylate as a colorless oily product (3.0 g, yield 92%). LCMS (m / z): 222.1 (M+H).

[0417] Step E: (3S,4S)-(4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methanol

[0418] Under ice bath conditions, 1 M LiAlH4-THF (17.63 mL, 17.63 mmol) was added dropwise to a solution of (3S,4S)-methyl 4-(difluoromethyl)-1,3-dimethylpiperidine-3-carboxylate (3.0 g, 13.56 mmol) in anhydrous THF (30 mL). The resulting mixture was stirred at 0 °C for 15 min. After the reaction was monitored by LCMS and completed, sodium sulfate decahydrate was added to quench the reaction until no bubbles were generated. Approximately 8 g of anhydrous sodium sulfate was added. The mixture was filtered through diatomaceous earth, and the filter cake was washed three times with anhydrous tetrahydrofuran. The filtrate was collected and concentrated to dryness to obtain (3S,4S)-(4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methanol as a colorless solid product (2.6 g, yield 99%). LCMS (m / z): 194.1 (M+H).

[0419] Intermediate b-d3

[0420]

[0421] (S,E)-(4-(Fluoromethylene)-3-methyl-1-(methyl-d3)piperidin-3-yl)methanol

[0422]

[0423] Step A: Methyl (S,E)-4-(fluoromethylene)-3-methyl-1-(methyl-d3)piperidine-3-carboxylate

[0424] At room temperature, potassium carbonate (5.55 g, 40.2 mmol) was added to a mixed solution of (S,E)-methyl 4-(fluoromethylene)-3-methylpiperidine-3-carboxylate hydrochloride (3.00 g, 13.4 mmol), iodomethane-d3 (2.33 g, 16.1 mmol) and ACN (100 mL). After addition, the mixture was heated to 90 °C and stirred overnight. After monitoring the reaction by LCMS until completion, the mixture was filtered, and the filter cake was rinsed with EA (50 mL). The filtrate was collected, concentrated and further purified by FCC (SiO2, EA / PE = 0 - 20%), to give (S,E)-methyl 4-(fluoromethylene)-3-methyl-1-(methyl-d3)piperidine-3-carboxylate (1.9 g, yield 69%) as a colorless liquid. LC-MS (m / z): 205.1 (M+H).

[0425] Step B: (S,E)-(4-(Fluoromethylene)-3-methyl-1-(methyl-d3)piperidin-3-yl)methanol

[0426] Under an ice bath, LiAlH4 (1 M in THF, 9.3 mmol, 9.3 mL) was added dropwise to a mixed solution of (S,E)-methyl 4-(fluoromethylene)-3-methyl-1-(methyl-d3)piperidine-3-carboxylate (1.9 g, 9.3 mmol) and THF (50 mL). After addition, the mixture was stirred in an ice bath at 0 °C for 0.5 h. After monitoring the reaction by LCMS until completion, the reaction was quenched with Na2SO4·10H2O until no gas was evolved. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated at low temperature (35 °C) to give 4-(fluoromethylene)-3-methyl-1-methyl-D3-piperidin-3-methanol (1.3 g, 79% yield) as a colorless liquid. LC-MS (m / z): 177.1 (M+H).

[0427] Intermediate b-d5

[0428]

[0429] (S,E)-(4-(Fluoromethylene)-3-methyl-1-(methyl-d3)piperidin-3-yl)methylene-d2-ol

[0430]

[0431] Step A: (S,E)-(4-(Fluoromethylene)-3-methyl-1-(methyl-d3)piperidin-3-yl)methylene-d2-ol

[0432] Under ice bath conditions, LiAlD4 powder (271.28 mg, 6.46 mmol) was added to a solution of methyl (S,E)-4-(fluoromethylene)-3-methyl-1-(methyl-d3)piperidine-3-carboxylate (1.10 g, 5.39 mmol) in anhydrous tetrahydrofuran (15 mL). The resulting mixture was stirred at room temperature for 15 min. After the reaction was monitored by LCMS and completed, the reaction solution was quenched with sodium sulfate decahydrate until no bubbles were generated. Then, about 5 g of anhydrous sodium sulfate was added to the reaction solution to remove water. The reaction solution was filtered through diatomaceous earth, and the filter cake was washed three times with anhydrous tetrahydrofuran. The filtrate was collected and concentrated to dryness to obtain a colorless oily product, (S,E)-(4-(fluoromethylene)-3-methyl-1-(methyl-d3)piperidin-3-yl)methylene-d2-ol (951 mg, yield 99%). This product was directly used in the next reaction without purification. LCMS (m / z): 179.1 (M+H).

[0433] Intermediate c-d3

[0434]

[0435] ((3S,4S)-4-(Difluoromethyl)-3-methyl-1-(methyl-d3)piperidin-3-yl)methanol

[0436]

[0437] The synthesis of Intermediate c-d3 was carried out with reference to the synthesis of Intermediate b-d3. Intermediate c-1-3 was used in Step A. LCMS (m / z): 197.1 (M+H).

[0438] Intermediate c-d5

[0439]

[0440] ((3S,4S)-4-(Difluoromethyl)-3-methyl-1-(methyl-d3)piperidin-3-yl)methy-d2-ol

[0441]

[0442] The synthesis of Intermediate c-d5 was carried out with reference to the synthesis of Intermediate b-d5. Intermediate c-1-3-d3 was used in Step A. LCMS (m / z): 199.1 (M+H).

[0443] The following intermediates were synthesized with reference to the above method.

[0444]

[0445] Intermediate L

[0446] (7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)boronic acid

[0447] Step A: 7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-ol

[0448] Under ice bath stirring, TIPSCl (177 g, 920 mmol) was added dropwise to a solution of 7-fluoro-8-((triisopropylsilyl)ethynyl)naphthalene-1,3-diol (CAS: 2621932-34-9, 300 g, 837 mmol) and imidazole (119 g, 1.76 mol) in DCM (3 L). After the addition was completed, the system was slowly warmed to room temperature and stirred for 6 h. The reaction was monitored by TLC. After completion, water (900 mL) was added, and the mixture was stirred for 30 min. The layers were separated. The aqueous phase was extracted with DCM (900 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by silica gel Plug (PE / EA = 50:1) to obtain the compound 7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-ol (397 g, yield 92%).

[0449] Step B: 7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl trifluoromethanesulfonate

[0450] Under the condition of -45 to -35 °C, trifluoromethanesulfonic anhydride (326 g, 1.16 mol) was added dropwise to a solution of 7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-ol (397 g, 0.77 mol) and DIPEA (298 g, 2.31 mol) in DCM (4 L). After the addition was completed, the temperature was maintained and the mixture was stirred for 0.5 h. The reaction was monitored by TLC. The system was added to water (800 mL), and the layers were separated. The aqueous phase was extracted with DCM (1.2 L). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by silica gel Plug (PE / EA = 50:1) to obtain the compound 7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl trifluoromethanesulfonate (469 g, yield 94%).

[0451] Step C: (7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)boronic acid

[0452] Under nitrogen protection, Pd(dppf)Cl2 (13.2 g, 18.2 mmol) was added to a solution of 7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl trifluoromethanesulfonate (235 g, 0.36 mol), 5,5,5',5'-tetramethyl-2,2'-di(1,3,2-dioxaborolane) (164 g, 0.73 mol) and potassium acetate (107 g, 1.1 mol) in dioxane (2.4 L). The system was heated to 85 °C and stirred for 20 h. The reaction was monitored by TLC until completion, cooled to room temperature, filtered through diatomaceous earth, rinsed with EA, concentrated and purified by silica gel column (EA / PE = 0 - 5%) to obtain the crude compound.

[0453] The above crude compound was dissolved in methanol (1.2 L), 1N HCl (2.4 L) was added, and the resulting mixture was stirred at room temperature for 30 min. EA (2.4 L) was added and stirring was continued for 2 h. The mixture was allowed to stand and separated, and the organic phase was washed successively with water (2.4 L) and saturated brine (2.4 L × 2), concentrated to obtain (7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)boronic acid (183 g, yield 92%). 1 1H NMR (400 MHz, Chloroform-d) δ 7.66–7.60 (m, 1H), 7.34 (d, J = 2.5 Hz, 1H), 7.24–7.19 (m, 1H), 7.18 (d, J = 2.5 Hz, 1H), 4.52 (s, 2H), 1.35–1.29 (m, 3H), 1.24–1.21 (m, 3H), 1.20–1.17 (m, 18H), 1.12 (d, J = 7.3 Hz, 18H). LCMS (m / z): 543.3 (M+H).

[0454] Example 1

[0455]

[0456] 4-(2-(((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]nonan-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethyl-6-fluoronaphthalen-2-ol

[0457]

[0458] Step A: 6-(7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0459] Under N2, 2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS: 262193248-5, 406 mg, 1.1 mmol), potassium phosphate (700 mg, 3.3 mmol) and cata CXium A Pd-G3 (240 mg, 0.33 mmol) were added to a solution of 6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (400 mg, 1.1 mmol) in 1,4-dioxane (10 mL) and water (2 mL). The resulting reaction mixture was stirred at 100 °C for 2 h. After monitoring the reaction by LCMS until completion, the reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated to dryness. The obtained crude product was purified by FCC (SiO2, EA / PE = 0 - 35%) to give the yellow solid product 6-(7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (420 mg, yield 67%). LCMS (m / z): 553.1 (M+H).

[0460] Step B: 6-(7-(8-Ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0461] At room temperature, m-chloroperoxybenzoic acid (154 mg, 0.76 mmol) was added to a solution of 6-(7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (420 mg, 0.76 mmol) in dichloromethane (10 mL), and the mixture was stirred at this temperature for 1 h. After monitoring the reaction by LCMS until completion, 50 mL of saturated sodium bicarbonate solution was added to the reaction mixture, and then the mixture was extracted with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness to obtain the crude product 6-(7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (380 mg). LCMS (m / z): 569.1 (M+H).

[0462] Step C: 6-(2-(((3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0463] At -40 °C, to a solution of 6-(7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (300 mg) and ((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methanol (132 mg, 0.69 mmol) in anhydrous tetrahydrofuran (6 mL) was added 1N sodium bis(trimethylsilyl)amide solution (0.22 mL, 0.22 mmol). The resulting reaction mixture was stirred at -40 °C for an additional 2 h. After completion of the reaction monitored by LCMS, the reaction mixture was poured into 30 mL of saturated ammonium chloride solution, extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product 6-(2-(((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (400 mg) as a yellow solid. LCMS (m / z): 698.3 (M+H).

[0464] Step D: 4-(2-(((3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethyl-6-fluoronaphthalen-2-ol

[0465] At room temperature, trifluoroacetic acid (2 mL) was added to 6-(2-(((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (300 mg), and the reaction mixture was stirred at this temperature for 0.5 h. After the reaction was monitored by LCMS and completed, the reaction solution was purified by pre-HPLC (C18, CAN / (10 mmol NH4HCO3 / H2O) = 55 - 75%) to obtain the white solid product 4-(2-(((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethyl-6-fluoronaphthalen-2-ol (45 mg, yield 14%). 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (s, 1H), 9.23 (s, 1H), 7.77 (dd, J = 9.1, 6.0 Hz, 1H), 7.40–7.30 (m, 2H), 7.05 (dd, J = 4.3, 2.7 Hz, 1H), 6.31 (t, J = 55.7 Hz, 1H), 4.53–4.12 (m, 6H), 4.08–3.85 (m, 1H), 3.61–3.41 (m, 1H), 2.89–2.74 (m, 2H), 2.46–2.31 (m, 3H), 2.19–2.04 (m, 5H), 1.93–1.58 (m, 8H), 1.12 (s, 3H), 0.79–0.69 (m, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -119.52~-119.65, -139.08~-139.27. LCMS (m / z): 654.3 (M+H).

[0466] Examples 2 and 3

[0467]

[0468] 4-(2-(((3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-((R)-1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethyl-6-fluoronaphthalen-2-ol and 4-(2-(((3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-((S)-1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethyl-6-fluoronaphthalen-2-ol

[0469]

[0470] Step A: 4-(2-(((3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethyl-6-fluoronaphthalen-2-ol (Example 1, 45 mg) was resolved by SFC (Waters SFC 150, REGIS (S,S) WHELK-O1 (250*40 mm 10 μm), Supercritical CO2 / EtOH (+0.1% 7.0 mol / l Ammonia in MeOH) = 80 / 20). The first eluted isomer was Example 2 (5 mg, with a relatively smaller retention time). Chiral analysis method SFC-1, Rt = 4.409 min. 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (s, 1H), 9.23 (s, 1H), 7.77 (dd, J = 9.1, 6.0 Hz, 1H), 7.41–7.29 (m, 2H), 7.12–6.99 (m, 1H), 6.31 (t, J = 56.0 Hz, 1H), 4.49–3.88 (m, 7H), 3.60–3.46 (m, 1H), 2.91–2.75 (m, 2H), 2.44–2.30 (m, 3H), 2.15–2.03 (m, 5H), 1.89–1.61 (m, 8H), 1.12 (s, 3H), 0.78–0.70 (m, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -115.43~-119.39, -119.58, -139.08~-139.25. LCMS (m / z): 654.3 (M+H). The subsequently eluted isomer was Example 3 (16 mg, with a relatively larger retention time). Chiral analysis method SFC-1, Rt = 5.262 min. 11H NMR (400 MHz, DMSO-d6) δ 9.95 (s, 1H), 9.22 (s, 1H), 7.77 (dd, J = 9.1, 6.0 Hz, 1H), 7.42–7.28 (m, 2H), 7.13–6.98 (m, 1H), 6.31 (t, J = 55.6 Hz, 1H), 4.56–3.85 (m, 7H), 3.65–3.45 (m, 1H), 2.88–2.78 (m, 2H), 2.45–2.32 (m, 3H), 2.16–2.05 (m, 5H), 1.88–1.59 (m, 8H), 1.12 (s, 3H), 0.79–0.66 (m, 3H). 19 19F NMR (376 MHz, DMSO-d6) δ -115.55~-119.41, -119.56~-119.60, -139.07~-139.28. LCMS (m / z): 654.3 (M+H).

[0471] Chiral analysis method SFC-1: Waters UPCC (CA-352), analytical column: REGIS (S,S) WHELK-O1 (100*3 mm*3 μm); mobile phase A: CO2, mobile phase B: EtOH (+0.1% 7.0 mol / l Ammonia in MeOH); flow rate: 1.5 mL / min; column temperature: 35 °C; back pressure: 1800 psi; gradient: 0 - 8.0 min A / B = 80 / 20.

[0472] Prepare and characterize the following compounds with reference to the above synthesis scheme and appropriate modifications:

[0473]

[0474]

[0475]

[0476]

[0477] Example 17

[0478]

[0479] 4-(2-(((3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-4-((S)-1-oxa-6-aza-6-spiro[3.5]nonyl)-6,8-difluoro-7-quinazolinyl)-5-ethyl-6-fluoro-2-naphthol

[0480]

[0481] Step A: 4-((S)-1-oxo-6-aza-6-spiro[3.5]nonyl)-7-(8-ethyl-7-fluoro-3-methoxymethoxy-1-naphthyl)-2,6,8-trifluoroquinazoline

[0482] Under N2, 2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (278 mg, 0.7 mmol), potassium phosphate (218 mg, 1.0 mmol) and cata CXium APd-G3 (111 mg, 0.15 mmol) were added to a solution of compound 4-((S)-1-oxo-6-aza-6-spiro[3.5]nonyl)-7-bromo-2,6,8-trifluoroquinazoline (200 mg, 0.5 mmol) in 1,4-dioxane (5 mL) and water (1 mL). The resulting reaction solution was stirred at 100 °C for 5 h. After monitoring the reaction by LCMS and completion of the reaction, the reaction solution was filtered through diatomaceous earth, the filtrate was concentrated to dryness, and the obtained crude product was purified by FCC (SiO2, EA / PE = 0 - 45%) to obtain the yellow solid product 4-((S)-1-oxo-6-aza-6-spiro[3.5]nonyl)-7-(8-ethyl-7-fluoro-3-methoxymethoxy-1-naphthyl)-2,6,8-trifluoroquinazoline (190 mg, yield 68%). LCMS (m / z): 542.2 (M+H).

[0483] Step B: 2-(((3S,4S)-4-(difluoromethyl)-1-methyl-3-methylpiperidin-3-yl)methoxy)-4-((S)-1-oxa-6-aza[3.5]nonyl)-7-(8-ethyl-7-fluoro-3-methoxymethoxy-1-naphthyl)-6,8-difluoroquinazoline

[0484] At 0 °C, NaH (12 mg, 0.30 mmol) was added to a solution of 4-((S)-1-oxa-6-aza-6-spiro[3.5]nonyl)-7-(8-ethyl-7-fluoro-3-methoxymethoxy-1-naphthalenyl)-2,6,8-trifluoroquinazoline (80 mg, 0.15 mmol) and ((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methanol (43 mg, 0.22 mmol) in anhydrous tetrahydrofuran (5 mL). The resulting reaction mixture was stirred at 0 °C for an additional 1 h. After completion of the reaction monitored by LCMS, the reaction mixture was poured into 30 mL of saturated ammonium chloride solution, extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The obtained crude product was purified by FCC (SiO2, EA / PE = 30 - 60%) to give 2-(((3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-4-((S)-1-oxa-6-aza[3.5]nonyl)-7-(8-ethyl-7-fluoro-3-methoxymethoxy-1-naphthalenyl)-6,8-difluoroquinazoline (70 mg, yield 66%). LCMS (m / z): 715.2 (M+H).

[0485] Step C: 4-(2-(((3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-4-((S)-1-oxa-6-aza-6-spiro[3.5]nonyl)-6,8-difluoro-7-quinazolinyl)-5-ethyl-6-fluoro-2-naphthol

[0486] At room temperature, TFA (1 mL) was added to a solution of 2-(((3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-4-((S)-1-oxa-6-aza[3.5]nonyl)-7-(8-ethyl-7-fluoro-3-methoxymethoxy-1-naphthalenyl)-6,8-difluoroquinazoline (70 mg, 0.1 mmol). The reaction mixture was stirred at this temperature for 0.5 h. After completion of the reaction monitored by LCMS, the reaction mixture was purified by pre-HPLC (C18, CAN / (10 mmol NH4HCO3 / H2O) = 55 - 75%) to give the white solid product 4-(2-(((3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-4-((S)-1-oxa-6-aza-6-spiro[3.5]nonyl)-6,8-difluoro-7-quinazolinyl)-5-ethyl-6-fluoro-2-naphthol (2 mg, yield 3%). LCMS (m / z): 671.3 (M+H).

[0487]

[0488]

[0489]

[0490] Example 27

[0491]

[0492] 4-(2-(((3S,4S)-4-(Difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethyl-6-fluoronaphthalen-2-ol

[0493]

[0494] Step A: 6-(8-Fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0495] Under N2, to a solution of compound 6-(7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (500 mg, 1.4 mmol) in 1,4-dioxane (10 mL) and water (2 mL) were added (7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)boronic acid (1.15 g, 2.1 mmol), potassium phosphate (897 mg, 4.2 mmol) and cata CXium A Pd-G3 (103 mg, 0.14 mmol). The resulting reaction solution was stirred at 100 °C for 5 h. After monitoring the reaction by LCMS until completion, the reaction solution was filtered through diatomaceous earth, the filtrate was concentrated to dryness, and the obtained crude product was purified by FCC (SiO2, EA / PE = 0 - 15%) to give the yellow solid product 6-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (370 mg, yield 32%). LCMS (m / z): 718.3 (M+H).

[0496] Step B: 6-(8-Fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0497] At room temperature, to a solution of 6-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (370 mg, 0.45 mmol) in dichloromethane (5 mL) was added meta-chloroperoxybenzoic acid (92 mg, 0.45 mmol), and the reaction was stirred at this temperature for 1 h. After monitoring the reaction by LCMS until completion, 20 mL of saturated sodium bicarbonate solution was added to the reaction solution, and then extracted with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness to obtain the crude product 6-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (380 mg). LCMS (m / z): 833.2 (M+H).

[0498] Step C: 6-(2-(((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane

[0499] At -40 °C, to a solution of 6-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (180 mg, 0.22 mmol) and ((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methanol (50 mg, 0.26 mmol) in anhydrous tetrahydrofuran (5 mL) was added 1N sodium bis(trimethylsilyl)amide solution (0.44 mL, 0.44 mmol). The resulting reaction mixture was stirred at -40 °C for an additional 1 h. After completion of the reaction monitored by LCMS, the reaction mixture was poured into 20 mL of saturated ammonium chloride solution. Then it was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness to give the crude product as a yellow solid, 6-(2-((((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (130 mg). LCMS (m / z): 962.3 (M+H).

[0500] Step D: 4-(2-((((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthalen-2-ol

[0501] At room temperature, CsF (205 mg, 1.3 mmol) was added to a solution of 6-(2-(((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-oxa-6-azaspiro[3.5]nonane (130 mg, 0.13 mol) in DMF (2 mL), and the mixture was stirred at 50 °C for 1 h. After the reaction was monitored by LCMS and completed, the reaction solution was purified by pre-HPLC (C18, CAN / (10 mmol NH4HCO3 / H2O) = 45 - 75%) to obtain the white solid product 4-(2-(((3S,4S)-4-(difluoromethyl)-1,3-dimethylpiperidin-3-yl)methoxy)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthalen-2-ol (32 mg, yield 36%). 1 H NMR (400 MHz, DMSO-d6) δ 10.16 (s, 1H), 9.29–9.06 (m, 1H), 7.98 (dd, J = 9.3, 5.8 Hz, 1H), 7.51–7.43 (m, 1H), 7.40 (d, J = 2.5 Hz, 1H), 7.21 (dd, J = 5.0, 2.5 Hz, 1H), 6.32 (t, J = 55.4 Hz, 1H), 4.52–4.10 (m, 6H), 4.04–3.80 (m, 2H), 3.58–3.41 (m, 1H), 3.30 (s, 1H), 2.91–2.76 (m, 2H), 2.44–2.31 (m, 1H), 2.21–2.00 (m, 4H), 1.97–1.55 (m, 8H), 1.13 (s, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -110.71, -140.03. LCMS (m / z): 650.3 (M + H).

[0502]

[0503]

[0504]

[0505]

[0506]

[0507]

[0508] Example 57

[0509]

[0510] 7'-([(3S,4S)-4-(Difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy)-8-ethyl-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3',6',8'-triaza-[1,2'-binaphthalen]-3-ol

[0511]

[0512] Step A: (2-(1',7-Difluoro-7'-(methylthio)-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6',8'-triaza-1,2'-binaphthalen-8-ylethynyl)tris(isopropyl)silane

[0513] Under room temperature conditions, after replacing the mixed solution of 7-chloro-8-fluoro-2-(methylthio)-4-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-1,3,6-triazaindole (1.83 g, 3.38 mmol), cataCXium A Pd G3 (274 mg, 376 μmol), K3PO4 (980 mg, 5.66 mmol) and 1,4-dioxane / H2O = 4 / 1 (15 mL) with N2 three times, the temperature was raised to 110 °C and stirred for 2 h. After the reaction was detected by LCMS to be completed, the reaction solution was poured into H2O (100 mL), extracted with EA (60 mL × 3), and the collected organic phase was washed with saturated NaCl (20 mL). After the organic solution was concentrated, the obtained crude product was purified by FCC ((EtOH / EA = 1 / 3) / PE = 0 - 20%), and a yellow solid (2-{1',7-difluoro-7'-(methylthio)-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6',8'-triaza-1,2'-binaphthalen-8-ylethynyl)tris(isopropyl)silane (900 mg, 59%) was obtained. LCMS (m / z): 804.2 (M+H).

[0514] Step B: (2-(1',7-Difluoro-7'-(methylsulfinyl)-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6'-,8'-triaza-1,2'-binaphthalen-8'-ylethynyl)tris(isopropyl)silane

[0515] At room temperature, m-chloroperoxybenzoic acid (273 mg, 1.34 mmol, 85% content) was added to a solution of 2-(1',7-difluoro-7'-(methylthio)-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6',8'-triaza-1,2'-binaphthalen-8-(ethynyl)tris(isopropanol)silane (900 mg, 1.34 mmol) and DCM (15 mL), and the mixture was stirred at room temperature for 2 h. After the reaction was monitored by TLC and LCMS and completed, the reaction solution was diluted with DCM (50 mL), washed with semi-saturated aq. NaHCO3 (20 mL), extracted with DCM (60 mL×3), the organic phase was collected, washed with saturated NaCl (30 mL), dried over anhydrous Na2SO4, filtered, and the organic solution was concentrated to obtain a yellow solid (2-{1',7-difluoro-7'-(methylsulfinyl)-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6'-,8'-triaza-1,2'-binaphthalen-8'-ethynyl)tris(isopropanol)silane (900 mg, yield 98%). LCMS (m / z): 819.2 (M+H).

[0516] Step C: [2-(7'-([(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy)-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6',8'-triaza-1,2'-binaphthalen-8-yl)ethynyl]tris(isopropanol)silane

[0517] At -78 °C, t-BuONa (0.73 mL, 2.0 M THF solution, 1.46 mmol) was added dropwise to a solution of [(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methanol (85 mg, 440 μmol) in THF (3 mL). After stirring for 0.5 h, (2-{1',7-difluoro-7'-(methylsulfinyl)-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6'-,8'-triaza-1,2'-binaphthalen-8'-ylethynyl)tris(isopropanol)silane (300 mg, 366 μmol) was added to the above reaction solution, and the mixture was stirred at -78 °C for 1 h. After monitoring the reaction by LCMS until completion, the reaction solution was poured into semi-saturated NH4Cl (50 mL), extracted with EA (50 mL x 3). After collecting the organic phase, it was washed with saturated NaCl (20 mL) solution, dried over anhydrous Na2SO4, concentrated, and the crude product was purified by FCC (SiO2, (EtOH / EA = 1 / 3) / PE = 0 - 40%) to obtain a yellow solid [2-(7'-([(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy)-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6',8'-triaza-1,2'-binaphthalen-8-yl)ethynyl]tris(isopropanol)silane (110 mg, yield 32%). LCMS (m / z): 474.8 (M / 2 + H).

[0518] Step D: 7'-{[(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy}-8-ethynyl-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3',6',8'-triaza-[1,2'-binaphthalenyl]-3-ol

[0519] At room temperature, CsF (100 mg, 0.66 mmol) was added to a solution of [2-(7'-([(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy)-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3-[tris(isopropyl)methoxy]-3',6',8'-triaza-[1,2'-binaphthalenyl]-8-yl)ethynyl]tris(isopropanol)silane (110 mg, 116 μmol) in DMF (3 mL). The reaction mixture was heated to 45 °C and stirred for 1 h. After completion of the reaction monitored by LCMS, the reaction solution was prepared by Pre-HPLC (C18, ACN / 10 mM NH4HCO3 = 50 - 80%), and a pale yellow solid, 7'-{[(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy}-8-ethynyl-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3',6',8'-triaza-[1,2'-binaphthalenyl]-3-ol (60 mg, yield 81%) was obtained. LCMS (m / z): 636.3 (M+H). 1 H NMR (400 MHz, Methanol-d4) δ 9.43–9.30 (m, 1H), 7.91–7.80 (m, 1H), 7.38–7.26 (m, 2H), 7.22 (d, J = 2.5 Hz, 1H), 6.20 (t, J = 55.6 Hz, 1H), 4.80–4.50 (m, 3H), 4.06–3.95 (m, 1H), 3.90–3.78 (m, 2H), 3.74–3.65 (m, 1H), 3.55–3.49 (m, 0.6H), 3.41–3.34 (m, 1H), 3.27–3.25 (m, 0.4H), 3.16–3.05 (m, 1H), 3.01–2.92 (m, 1H), 2.49–2.32 (m, 1H), 2.24 (s, 3H), 2.05–1.70 (m, 6H), 1.47–1.38 (m, 1H), 1.32–1.27 (m, 1H), 1.24–1.16 (m, 3H), 0.96–0.75 (m, 1H). 19 F NMR (376 MHz, Methanol-d4) δ -110.94–-113.36, -118.79–-122.50, -139.31–-140.89。

[0520] Step E: 7'-([(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy)-8-ethyl-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3',6',8'-triaza-[1,2'-binaphthalen]-3-ol

[0521] At room temperature, 7'-([(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy)-8-ethynyl-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3',6',8'-triaza-[1,2'-binaphthalenyl]-3-ol (30 mg, 47.2 μmol) and Pd / C (28 mg, 10% w / w, 23.6 μmol) were dissolved in methanol (10 mL). The air was displaced twice with an H2 balloon, and the mixture was stirred at room temperature under an H2 balloon for 1 h. After monitoring the reaction by LCMS until completion, the reaction solution was filtered to obtain a clear organic phase, which was concentrated completely. Acetonitrile (1 mL) and deionized water (2 mL) were added, and the mixture was freeze-dried to obtain a white solid, 7'-([(3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl]methoxy)-8-ethyl-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3',6',8'-triaza-[1,2'-binaphthalene]-3-ol (20 mg, yield 66%). LCMS (m / z): 640.3 (M+H). 1 1H NMR (400 MHz, Methanol-d4) δ 9.46–9.33 (m, 1H), 7.67 (dd, J = 9.0, 5.9 Hz, 1H), 7.29 (d, J = 2.7 Hz, 1H), 7.27–7.19 (m, 1H), 7.11–7.04 (m, 1H), 6.20 (t, J = 55.6 Hz, 1H), 4.76–4.49 (m, 3H), 4.05–3.94 (m, 1H), 3.89–3.78 (m, 2H), 3.75–3.63 (m, 1H), 3.43–3.34 (m, 1H), 3.15–3.04 (m, 1H), 3.00–2.90 (m, 1H), 2.53–2.33 (m, 2H), 2.25–2.15 (m, 4H), 1.98–1.72 (m, 6H), 1.45–1.36 (m, 1H), 1.25–1.17 (m, 3H), 0.87–0.68 (m, 4H). 19 19F NMR (376 MHz, Methanol-d4) δ -117.86– -123.89, -136.19– -142.09。

[0522] Prepare and characterize the following compounds according to the above synthetic scheme and appropriate modifications.

[0523]

[0524]

[0525]

[0526]

[0527] Example 81

[0528]

[0529] 7'-(((3S,4S)-4-(difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-8-ethynyl-1',7-difluoro-5'-(2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-3',6',8'-triaza-[1,2'-binaphthalen]-3-ol

[0530]

[0531] For the synthesis of Example 81, see the relevant steps described in the synthesis of Example 57. LCMS (m / z): 636.3 (M+H). 1 HNMR (400 MHz, Methanol-d4) δ 9.43–9.30 (m, 1H), 7.91–7.80 (m, 1H), 7.38–7.26 (m, 2H), 7.22 (d, J = 2.5 Hz, 1H), 6.20 (t, J = 55.6 Hz, 1H), 4.80–4.50 (m, 3H), 4.06–3.95 (m, 1H), 3.90–3.78 (m, 2H), 3.74–3.65 (m, 1H), 3.55–3.49 (m, 0.6H), 3.41–3.34 (m, 1H), 3.27–3.25 (m, 0.4H), 3.16–3.05 (m, 1H), 3.01–2.92 (m, 1H), 2.49–2.32 (m, 1H), 2.24 (s, 3H), 2.05–1.70 (m, 6H), 1.47–1.38 (m, 1H), 1.32–1.27 (m, 1H), 1.24–1.16 (m, 3H), 0.96–0.75 (m, 1H). 19 F NMR (376 MHz, Methanol-d4) δ -110.94–-113.36, -118.79–-122.50, -139.31–-140.89.

[0532] Example 82

[0533]

[0534] 7'-(((3S,4S)-4-(Difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-5'-((1S,7S)-2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-8-ethynyl-1',7-difluoro-3',6',8'-triaza-(1,2'-binaphthyl)-3-ol

[0535]

[0536] The synthesis of Example 82 was carried out with reference to Example 57, using Intermediate A-II-A in Step A. LCMS (m / z): 636.3 (M+H). 1 H NMR (400 MHz, Methanol-d4) δ 9.47–9.30 (m, 1H), 7.96–7.73 (m, 1H), 7.43–7.29 (m, 2H), 7.26–7.16 (m, 1H), 6.47–5.97 (m, 1H), 4.59–4.49 (m, 3H), 4.08–3.95 (m, 1H), 3.90–3.78 (m, 2H), 3.76–3.65 (m, 1H), 3.54–3.47 (m, 0.5H), 3.20–3.06 (m, 1H), 3.03–2.92 (m, 1H), 2.60–2.41 (m, 1H), 2.24 (s, 3H), 2.21–2.15 (m, 0.5H), 2.07–1.96 (m, 2H), 1.95–1.86 (m, 1H), 1.83–1.73 (m, 3H), 1.66–1.52 (m, 0.5H), 1.48–1.39 (m, 1H), 1.25–1.17 (m, 3H), 0.96–0.87 (m, 1H), 0.84–0.75 (m, 0.5H). 19 F NMR (376 MHz, Methanol-d4) δ -111.71, -119.23, -121.69, -140.14.

[0537] Example 83

[0538]

[0539] 7'-(((3S,4S)-4-(Difluoromethyl)-1-methyl-3-methyl-3-piperidinyl)methoxy)-5'-((1S,7S)-2-oxa-6-azabicyclo[5.1.0]oct-6-yl)-8-ethynyl-1',7-difluoro-3',6',8'-triaza-(1,2'-binaphthyl)-3-ol

[0540]

[0541] The synthesis of Example 83 was carried out with reference to Example 57, using intermediate A-II-B in Step A. LCMS (m / z): 636.3 (M+H). 1 H NMR (400 MHz, Methanol-d4) δ 9.46–9.30 (m, 1H), 7.94–7.75 (m, 1H), 7.44–7.27 (m, 2H), 7.21 (d, J = 2.6 Hz, 1H), 6.43–6.00 (m, 1H), 4.78–4.70 (m, 1H), 4.63–4.50 (m, 2H), 4.08–3.94 (m, 1H), 3.90–3.79 (m, 2H), 3.76–3.65 (m, 1H), 3.57–3.50 (m, 0.5H), 3.43–3.35 (m, 1H), 3.27–3.23 (m, 0.5H), 3.17–3.08 (m, 1H), 3.01–2.91 (m, 1H), 2.50–2.31 (m, 1H), 2.29–2.16 (m, 3H), 2.05–1.89 (m, 2H), 1.87–1.70 (m, 4H), 1.47–1.38 (m, 1H), 1.24–1.17 (m, 3H), 0.94–0.76 (m, 1H). 19 F NMR (376 MHz, Methanol-d4) δ -111.77, -119.41, -121.48, -140.13.

[0542] Synthesize and characterize the following compounds with reference to the above synthetic scheme and appropriate variants.

[0543]

[0544]

[0545]

[0546]

[0547]

[0548]

[0549]

[0550]

[0551] Active Examples

[0552] Example 1: Proliferation Inhibition Effect of the Compound of the Invention on NCI-H727 Cells with KRAS G12V Mutation

[0553] This experiment evaluated and verified the proliferation inhibitory activity of the compound of the invention against NCI-H727 cells with KRAS G12V mutation.

[0554] The NCI-H727 cells (Nanjing Kebai Biotechnology Co., Ltd., product number CBP60182, adherent, medium RPMI-1640 + 10% FBS (GIBCO, Cat#10091-148)) were cultured under the conditions of 37 °C, 5% CO2, and 95% humidity.

[0555] 3D Cell Viability Assay: Cells in the logarithmic growth phase were harvested and counted using a platelet counter. The cell viability was detected by trypan blue exclusion method to ensure that the cell viability was above 90%. RPMI-1640 medium containing 1% MC (Sigma, Cat#M0512) was prepared and 10% FBS was added to prepare 3D cell medium. The cell concentration was adjusted to 14,815 cells / mL with 3D medium and the content of MC was 0.65%; 135 μL of cell suspension was added to each well of a 96-well clear flat bottom black wall plate (Greiner, Cat#655096); the cells in the 96-well plate were cultured overnight under the conditions of 37 °C and 5% CO2.

[0556] IC50 Determination Drug Preparation: A 10-fold drug solution was prepared with the medium. 10 μL of the drug solution was added to each well of the 96-well plate inoculated with cells, so that the working concentration was up to 10 μM, diluted 3-fold, 9 concentrations, and 2 replicates were set for each drug concentration. The cells in the 96-well plate with added drug were continuously cultured under the conditions of 37 °C and 5% CO2 for 7 days, and then CTG detection ( Luminescent Cell Viability Assay (Promega, Cat#G7573)).

[0557] Equilibrate the cell plate to room temperature for 30 minutes and thaw the CTG reagent ( Luminescent Cell Viability Assay (Promega, Cat# G7573)), add 75 μL of CTG solution to each well for detection, and shake on an orbital shaker for 5 minutes to lyse the cells. Place the cell plate at room temperature for 25 minutes to stabilize the bioluminescence signal, and read the bioluminescence value ( Multifunctional microplate reader, PerkinElmer #2105).

[0558] Use GraphPad Prism software to analyze the data, fit the data using the Dose-response-inhibition equation to obtain the dose-effect curve, and calculate the IC50 value therefrom.

[0559] Cell viability (%) = (Lum of test drug - Lum of culture medium control) / (Lum of cell control - Lum of culture medium control) × 100%.

[0560] The compounds of the present invention show satisfactory anti-proliferative activity against NCI-H727 human lung cancer cells with KRAS G12V mutation, and the IC50 ranges from <1000 nM, preferably <100 nM. Representative activity data are shown in the following table:

[0561] Examples <![CDATA[IC 50 (nM) / NCI-H727 <!-- 86 -->]]> Example 1 18 Example 3 4.2 Example 4 16 Example 19 42 Example 27 140 Example 57 29 Example 81 12 Example 83 8.2

[0562] Example 2: Proliferation inhibitory effect of the compounds of the present invention on AGS cells with KRAS G12D mutation

[0563] This experiment evaluated and verified the proliferation inhibitory activity of the compounds of the present invention against AGS cells with KRAS G12D mutation.

[0564] Culture AGS cells (Nanjing Kebai Biotechnology Co., Ltd., product number CBP60476, adherent, medium (F12K Nutrient Mixture + 10% FBS (GIBCO, Cat# 10091-148)) at 37 °C, 5% CO2, 95% humidity conditions, 1500 / well, harvest cells in the logarithmic growth phase and use a Countstar automatic cell counter based on the principle of classical trypan blue staining method to perform cell counting and detect cell viability to ensure that the cell viability is above 90%. Adjust the cell concentration; add 80 μL of cell suspension to a 96-well clear flat-bottom black-wall plate (Greiner, Cat# 655090) respectively, and place the cells in the 96-well plate at 37 °C, 5% CO2 conditions for culture.

[0565] IC 50Drug preparation for measurement: Prepare a 5-fold drug solution with a culture medium. Add 20 μL of the drug solution to each well in a 96-well plate inoculated with cells, such that the working concentration is up to 10 μM, diluted 3-fold, with 9 concentrations, and set 2 replicates for each drug concentration. Incubate the cells in the 96-well plate with added drugs at 37 °C and 5% CO2 for another 3 days, and then perform CTG detection.

[0566] Equilibrate the cell plate to room temperature for 30 minutes and thaw the CTG reagent ( Luminescent CellViability Assay (Promega, Cat#G7573)). Add 50 μL of the CTG solution to each well, and shake on an orbital shaker for 2 minutes to lyse the cells. Place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal, and read the luminescence value ( Multifunctional microplate reader, PerkinElmer#2105).

[0567] Use GraphPad Prism software to analyze the data, fit the data with the Dose-response-inhibition equation to obtain the dose-effect curve, and calculate the IC50 value therefrom.

[0568] Cell viability (%) = (Lum 待测药 -Lum 培养液对照 ) / (Lum 细胞对照 -Lum 培养液对照 ) × 100%.

[0569] The compounds of the present invention show satisfactory anti-proliferative activity against AGS human gastric adenocarcinoma cells with KRAS G12D mutation, with the IC50 range being <1000 nM, preferably <100 nM. Representative activity data are shown in the following table.

[0570] Example <![CDATA[IC 50 (nM) / AGS]]> Example 1 9.5 Example 3 6.7 Example 4 17.8 Example 19 61 Example 27 56 Example 57 41 Example 81 7.4 Example 83 11 .

[0571] Example 3: Rat cassette pharmacokinetic properties of the compounds of the present invention

[0572] The pharmacokinetic characteristics of the compounds of the present invention were evaluated through a rat cassette pharmacokinetic experiment (Nagilla R. et al., J. Pharm. Sci. 2011, 100, 3862–3874.).

[0573] Male Sprague-Dawley (SD) rats, 6-8 weeks old and weighing 220-250 g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The following reagents were used: Tolbutamide (Aladdin, product number H1401054); Sulfobutyl-β-cyclodextrin (Captisol, Shandong Binzhou Zhiyuan Biotechnology Co., Ltd., product number 20191013); Propylene glycol (15) stearate (Solutol, Meilun Biotechnology Co., Ltd., product number S0206A); DMSO (Vetec Co., product number WXBD0293V); Acetonitrile (Sigma-Aldrich, product number WXBD1744V); Methanol (Sigma-Aldrich, product number WXBD2831V).

[0574] The compound combination was formulated into a solvent of 5% DMSO / 10% Solutol / 85% (20% Captisol), and the final concentration of each compound was 1 mg / mL. The drug formulation was injected into the tail vein of SD rats at an injection volume of 1 mL / kg. Blood samples were collected from the external jugular vein at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h, centrifuged at low temperature for 20 minutes, and the plasma was collected and stored at -20°C for further analysis.

[0575] The following LC-MS / MS analytical method for the compound was established:

[0576] Standard curve preparation: 20 μL of 1 mg / mL DMSO stock solution of each compound was pipetted and transferred into 900 μL of 50% methanol working solution, and serially diluted to obtain a standard curve working solution with concentrations of 20000, 10000, 5000, 1000, 500, 100, 50, 20, 10 ng / mL. Then, 5 μL of the standard curve working solution was mixed with 45 μL of rat blank plasma to obtain a standard curve with concentrations of 2000, 1000, 500, 100, 50, 10, 5, 2, 1 ng / mL for quantifying unknown samples.

[0577] Sample pretreatment: 50 μL of unknown plasma samples and standard curve samples were added with 250 μL of acetonitrile containing Tolbutamide as an internal standard as a precipitant to precipitate plasma proteins and extract the compounds to be measured in the plasma. After centrifugation at low temperature for 20 minutes, the supernatant was taken, mixed with an aqueous solution of 0.1% formic acid, and 5 μL was injected for LC-MS analysis of the drug plasma concentration.

[0578] The standard curve was plotted using the mass spectrometry analysis software Analyst 1.6.1 to quantify unknown samples, and the pharmacokinetic parameters were calculated using Winnonlin 8.2 based on the drug concentrations at each time point of the unknown samples.

[0579] The experimental results showed that in the pharmacokinetic evaluation of cassette dosing, the compounds of the present invention exhibited good pharmacokinetic properties.

[0580] Example 4: Inhibition Test of Compounds of the Present Invention on Cytochrome P450

[0581] This experiment evaluated the inhibitory effect of the inventive compounds on cytochrome P450.

[0582] This experiment was carried out using the following reagents: human liver microsomes (Corning, catalog number 452161); reduced nicotinamide adenine dinucleotide phosphate (NADPH, MCE, catalog number HY-F0003 / CS-4998); phenacetin, diclofenac, α-naphthoflavone, omeprazole and ketoconazole were all purchased from TCI; S-mephenytoin and testosterone were purchased from CAYMAN; midazolam was purchased from Bioreclamation IVT; quinidine was purchased from Damas-beta; sulfaphenazole was purchased from MCE; bufuralol was purchased from TRC.

[0583] 100 mM potassium phosphate buffer (K-buffer) was prepared with potassium dihydrogen phosphate and dipotassium hydrogen phosphate, and the pH was adjusted to 7.4 to prepare 0.1 M potassium phosphate buffer (K-buffer).

[0584] 8 μL of 10 mM test compound stock solution was dissolved in 12 μL of acetonitrile to prepare 400× test compound. 12 μL of 1 mM α-naphthoflavone, 10 μL of 40 mM sulfaphenazole, 10 μL of 10 mM quinidine and 8 μL of DMSO solution were mixed to prepare a mixed solution of CYP1A2, CYP2C9 and CYP2D6 inhibitors. 8 μL of 2.5 mM ketoconazole DMSO solution was dissolved in 12 μL of acetonitrile to prepare an inhibitor solution of CYP3A4; 8 μL of 100 mM omeprazole DMSO solution was dissolved in 12 μL of acetonitrile to prepare an inhibitor solution of CYP2C19.

[0585] 66.7 mg of NADPH was added to 10 mL of 0.1 M K-buffer, pH 7.4, to prepare 4× NADPH potassium phosphate solution. Each substrate was prepared into a solution required for 4-fold concentration determination with 10 mL of 0.1 M K-buffer according to the concentration requirements to obtain 4× substrate potassium phosphate solution.

[0586] 10 μL of 20 mg / mL human liver microsomes was added to 990 μL of K-buffer to prepare 0.2 mg / mL human liver microsomes (HLM) solution, which was stored on ice for later use.

[0587] Add 600 μL of 0.2 mg / mL HLM into a 96-well plate, and then add 3 μL of a 400-fold solution of the test compound; add 200 μL of 0.2 mg / mL HLM into a 96-well plate, and then add 1 μL of the diluted positive control inhibitor solution. Aliquot 30 μL of the mixed solution of the compound and human liver microsomes into a 96-well plate, and then add 15 μL of the substrate solution. Preheat the above-obtained solution and the prepared NADPH solution at 37 °C for 5 min. Add 15 μL of the preheated NADPH solution to the reaction plate, mix well, and start the reaction. Incubate the reaction plate at 37 °C. Incubate for 5 minutes for 3A4; incubate for 10 minutes for 1A2, 2C9, and 2D6; incubate for 45 minutes for 2C19. At the end of the reaction, add 120 μL of acetonitrile containing an internal standard to terminate the reaction. Vortex the sample for 10 min, centrifuge at 5594 g for 15 minutes, and send the prepared sample for LC-MS / MS analysis.

[0588] The experimental results show that at the tested concentrations, the compounds of the present invention have no significant inhibitory effect on the key CYP subtypes of drug metabolism, showing better drug-drug interaction safety.

[0589] Example 5: Proliferation inhibition effect of the compound of the present invention on NCI-H358 cells with KRAS G12C mutation

[0590] This experiment evaluated and verified the proliferation inhibitory activity of the compounds of the present invention on NCI-H358 cells with KRAS G12C mutation.

[0591] Culture NCI-H358 cells (Nanjing Kebai Biotechnology Co., Ltd., product number CBP60136, adherent, medium RPMI-1640 + 10% FBS (GIBCO, Cat#10091-148)) under the conditions of 37 °C, 5% CO2, and 95% humidity.

[0592] 3D cell viability detection: Harvest the cells in the logarithmic growth phase and count the cells using a platelet counter. Detect the cell viability by trypan blue exclusion method to ensure that the cell viability is above 90%. Prepare RPMI-1640 medium containing 1% MC (Sigma, Cat#M0512) and add 10% FBS to prepare a complete 3D cell medium. Adjust the cell concentration to 12,500 cells / mL with the 3D medium and make the content of MC 0.65%; add 80 μL of the cell suspension to a 96-well clear flat-bottom black-wall plate (Greiner, Cat#655096) respectively; incubate the cells in the 96-well plate overnight at 37 °C and 5% CO2.

[0593] IC50 determination drug preparation: Prepare a 5-fold drug solution with culture medium. Add 20 μL of the drug solution to each well of a 96-well plate inoculated with cells, such that the working concentration is up to 10 μM, diluted 3-fold, with 9 concentrations, and set 2 replicates for each drug concentration. Incubate the cells in the drug-added 96-well plate at 37 °C and 5% CO2 for another 5 days, and then perform CTG detection ( Luminescent Cell Viability Assay (Promega, Cat#G7573)).

[0594] Equilibrate the cell plate to room temperature for 30 minutes and thaw the CTG reagent ( Luminescent CellViability Assay (Promega, Cat#G7573)), add 50 μL of the CTG solution to each well for detection, and vibrate on an orbital shaker for 5 minutes to lyse the cells. Place the cell plate at room temperature for 25 minutes to stabilize the luminescence signal, and read the luminescence value ( Multifunctional microplate reader, PerkinElmer#2105).

[0595] Use GraphPad Prism software to analyze the data, fit the data with the Dose-response-inhibition equation to obtain the dose-effect curve, and calculate the IC50 value therefrom.

[0596] Cell viability (%) = (Lum 待测药 -Lum 培养液对照 ) / (Lum 细胞对照 -Lum 培养液对照 ) × 100%.

[0597] The compounds of the present invention show satisfactory anti-proliferative activity against NCI-H358 human lung cancer cells with KRAS G12C mutation, with the IC50 range being <1000 nM, preferably <100 nM. Representative activity data are shown in the following table.

[0598] Example <![CDATA[IC 50 (nM) / H358]]> Example 3 1.3 Example 4 4.2

[0599] Example 6: Proliferation inhibitory effect of the compounds of the present invention on 6 tumor cell lines

[0600] This experiment evaluated and verified the proliferation inhibitory activity of the compounds of the present invention against the following 6 KRAS-related tumor cell lines.

[0601] KRAS Cell Lines Cell Growth Characteristics Complete Medium G12V NCI-H441 Adherent RPMI-1640 + 10% FBS G12V Capan-2 Adherent RPMI-1640 + 10% FBS G12S A549 Adherent DMEM + 10% FBS G13D HCT116 Adherent RPMI-1640 + 10% FBS Q61H NCI-H460 Adherent RPMI-1640 + 10% FBS WT, Amplification EBC-1 Adherent RPMI-1640 + 10% FBS WT A375 Adherent DMEM + 10% FBS

[0602] The following materials, reagents and instruments were used in this experiment: 6 cell lines were all from Kangyuan Botech (Beijing) Co., Ltd. (NCI-H441, KC-0510; Capan-2, KC-0185; A549, KC-0284; HCT116, KC-0281; NCI-H460, KC-0512; EBC-1, KC-0195; A375, KC-0158); RPMI-1640 (Hyclone, SH30809.01); fetal bovine serum FBS (GIBCO, 10099-141); methylcellulose (SIGMA, 9004-67-5); phosphate buffer PBS (Solarbio, P1020-500); CellCounting-Lite 2.0 Luminescent Cell Viability Assay (Nanjing Novoprotein, DD1101-04); 96-well clear flat-bottom black-wall plates (Thermo, 165305); multi-functional microplate reader (BMGLABTECH, Plus); CO2 incubator (Thermo Scientific, Model 3100 Series).

[0603] Prepare sterilized 1% methylcellulose 3D medium in advance. Harvest cells in the logarithmic growth phase and count the cells using a hemocytometer. Detect cell viability by trypan blue exclusion method to ensure that the cell viability is above 90%. Adjust the cell concentrations of NCI-H441, A549, HCT116, NCI-H460, EBC-1 and A375, etc., so that the final concentration of methylcellulose is 0.65%, mix well and let stand; after there is no visible air in the cell suspension, add 180 μL of cell suspension to each well of a 96-well plate, with a total of 2500 cells. Adjust the capan-2 cell concentration using complete medium, and add 180 μL of cell suspension to each well of a 96-well plate, with a total of 3000 cells. Incubate the cells in the 96-well plate overnight at 37 °C, 5% CO2 and 95% humidity.

[0604] IC 50 Drug preparation for determination: First, prepare 10 mM DMSO stock solutions of each compound. For the first time, use DMSO as the solvent and dilute 3.16-fold to prepare DMSO solutions of different concentrations of each compound at 9 concentrations. For the second time, dilute 1:100 to prepare 10-fold diluted solutions of each compound, using complete medium as the solvent. Finally, add 20 μL of each compound diluted solution to each well of the 96-well plate seeded with cells. The final highest drug concentration is 10 μM, with 9 concentrations and 3.16-fold dilution. Each concentration is set with three replicates.

[0605] Place the cells in the 96-well plate with added drugs in an incubator at 37 °C and 5% CO2 for continuous culture for 144 hours, and then perform CTG analysis. Melt the CTG reagent and equilibrate the cell plate to room temperature for 30 minutes. Add an equal volume of CTG solution to each well. Vortex on an orbital shaker for 5 minutes to lyse the cells. Place the cell plate at room temperature for 20 minutes to stabilize the luminescence signal. Read the luminescence value and collect the data.

[0606] Analyze the data using GraphPad Prism 7.0 software, and use non-linear S-curve regression to fit the data to obtain a dose-effect curve, and calculate the IC 50 value.

[0607] Cell viability (%) = (Lum 待测药 -Lum 培养液对照 ) / (Lum 溶剂对照 -Lum 培养液对照 ) × 100%.

[0608] The compounds of the present invention, such as the compounds of the examples, show satisfactory anti-proliferative activity against KRAS mutant cells and wild amplified cells in this example, with the IC50 range being <1000 nM, preferably <500 nM. Representative activity data are shown in the following table.

[0609]

[0610] It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. These changes and modifications can be made without departing from the essence and scope of the subject matter of the present invention and without diminishing its intended advantages. Therefore, such changes and modifications are intended to be covered by the appended claims.

[0611] All published publications cited in this specification are incorporated herein by reference.

Claims

1. A compound of formula (I), its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein: M is selected from N or C-R9; W is selected from N or C-R 10 ; R9 is selected from H, halogen, CN, NO2 and -C 1-6 alkyl optionally substituted by halogen; R 10 selected from H, halogen, CN, OH, -C 1-6 alkyl optionally substituted with halogen and -OC 1-6 alkyl; Z is selected from H, OH and NH2; X is selected from CH2 and O, provided that when k is 0, X is CH2; Y is selected from O, S, Se and N-R a ; R1 is selected from H and optionally substituted -C 1-6 alkyl, wherein the substituent is selected from halogen, D and -O-C 1-6 alkyl optionally substituted by halogen or D; Each R2 is independently selected from H, D, and optionally substituted -C 1-6 alkyl, wherein the substituent is selected from halogen, D, and -O-C 1-6 alkyl optionally substituted by halogen or D; R3 is selected from H, D, halogen, -CN, -OH, -NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -O-C 1-6 alkyl, -O-C 3-6 cycloalkyl, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl, -(CH2) n -C 3-6 cycloalkyl and =C(R c )2, wherein each occurrence of C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl is optionally substituted, and the substituents are selected from halogen, CN, D and -OC 1-6 alkyl optionally substituted by halogen, wherein the C 3-6 cycloalkyl is optionally substituted, and the substituents are selected from halogen, CN, D, -C 1-6 alkyl optionally substituted by halogen and -OC 1-6 alkyl optionally substituted by halogen, or Two R3 groups attached to the same ring carbon atom, together with the carbon atom to which they are attached, form a spiro C 3-6 cycloalkyl or spiro 4-7-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from N, O, S, said cycloalkyl or heterocycloalkyl being optionally substituted by halogen or -C 1-6 alkyl substituted by halogen; R4 is selected from H, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl and -(CH2) n -C 3-6 cycloalkyl, wherein the -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl is optionally substituted, and the substituents are selected from D, halogen, CN, OH, -O-C 1-6 alkyl optionally substituted by halogen or D and -OCON(R b )2, wherein the C 3-6 cycloalkyl is optionally substituted, and the substituents are selected from D, halogen, CN, OH, -C 1-6 alkyl optionally substituted by halogen or D, -O-C 1-6 alkyl optionally substituted by halogen or D and -OCON(R b )2; R5 is selected from H, halogen, -CN, -NO2; R6 is selected from halogen, CN, -C 1-6 alkyl and -C 2-6 alkynyl, wherein the -C 1-6 alkyl and -C 2-6 alkynyl are each independently optionally substituted by halogen; R7 is selected from H, halogen, CN, -C 1-6 alkyl optionally substituted by halogen or D, -OC 1-6 alkyl and -C 2-6 alkynyl optionally substituted by halogen or D; R8 and R8 attached to non-adjacent ring carbon atoms ’ together form an in-ring bridging -(CH2) 1-2 - or -CH2=CH2-, or R8 and R8 attached to the same ring carbon atom ’ together with the ring carbon atom to which they are attached form a 4- to 6-membered spiroalkyl group or a 4- to 6-membered spiroheteroalkyl group containing 1 to 3 heteroatoms independently selected from N, O, and S, or R8 and R8' attached to adjacent ring carbon atoms together with the ring carbon atoms to which they are attached form a fused C 3-6 cycloalkyl or a 4-6 membered fused heterocycloalkyl containing 1 to 2 heteroatoms independently selected from N, O, and S, wherein the bridged ring, spiro ring or fused ring is each independently optionally substituted, and the substituent is selected from OH, oxo, -OC optionally substituted by halogen 1-6 alkyl and -C optionally substituted by halogen 1-6 alkyl; R a and R b each independently selected from H and optionally halogen-substituted -C 1-6 alkyl; R c each independently selected from H, halogen, and -C 1-6 alkyl optionally substituted by halogen; k and n are each independently selected from integers from 0 to 3; and m is selected from integers from 0 to 6.

2. The compound of claim 1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein M is N.

3. The compound of claim 1, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein M is C-F, C-Cl, C-CN, C-NO2, C-CF3.

4. The compound of any one of claims 1 to 3, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein W is C-halogen, preferably C-F.

5. A compound according to any one of claims 1 to 4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R7 is selected from H, -OC optionally substituted by halogen or D 1-6 alkyl and -C optionally substituted by halogen or D 2-6 alkynyl.

6. The compound of any one of claims 1 to 5, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R5 is selected from H and halogen, preferably F.

7. A compound according to any one of claims 1 to 6, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R6 is selected from halogen, -C 1-3 alkyl and -C 2-4 alkynyl, preferably F, Cl, ethyl and ethynyl.

8. A compound according to any one of claims 1 to 4, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein Z is OH, R5 is selected from halogen, and R6 is selected from halogen, -C 1-3 alkyl and -C 2-4 alkynyl.

9. The compound according to any one of claims 1 to 8, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment is selected from 10. The compound of claim 9, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment is selected from and a 5- to 7-membered heterocycloalkyl having an in-ring bridging -(CH2) 1-2 - or -CH2=CH2-, preferably -(CH2) 1-2 -, wherein G1 is selected from CH2, NH, O and S, and at least one of G2, G3, G4 is selected from CH2, NH, O and S, and the rest are each independently optionally selected from absent, CH2, NH, O and S; wherein the spiro ring or bridged ring is optionally substituted, and the substituents are selected from OH, oxo, -OC 1-6 alkyl optionally substituted by halogen and -C 1-6 alkyl optionally substituted by halogen.

11. The compound of claim 10, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein is wherein both G1 and G2 are CH2, or one of them is CH2 and the other is selected from NH, O and S, preferably NH and O; or is wherein G1, G2 and G3 are each independently selected from CH2, NH, O and S; wherein the spiro ring is optionally substituted, and the substituents are selected from OH, oxo, -OC 1-6 alkyl optionally substituted by halogen and -C 1-6 alkyl optionally substituted by halogen.

12. A compound according to any one of claims 1 to 11, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein, wherein Y is O.

13. A compound according to any one of claims 1 to 12, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural moiety is preferably 14. The compound of claim 13, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R2 is selected from H or D, for example, both R2 are H; or both R2 are D; or one of R2 is H and the other is D.

15. The compound of claim 13, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R1 is -C 1-3 alkyl or -deuterated C 1-3 alkyl.

16. The compound of claim 13, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R3 is selected from halogen, preferably F; -C 1-6 alkyl, preferably C 1-3 alkyl, substituted by halogen, preferably substituted by F; or =C(R c )2, wherein R c are each independently selected from H and halogen, preferably H and F; preferably, R3 is substituted at the para position of the ring N atom, and m is an integer from 1 to 2.

17. The compound of claim 13, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein R4 is selected from -C 1-3 alkyl and -deuterated C 1-3 alkyl, preferably -CH3, -CD3, -CH2CH3.

18. A compound according to any one of claims 1 to 11, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, wherein the structural fragment is selected from 19. A compound selected from the compounds of the examples or its pharmaceutically acceptable salt or solvate.

20. A pharmaceutical composition comprising the compound of any one of claims 1 - 19, its stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates, and a pharmaceutically acceptable excipient.

21. The compound of any one of claims 1 - 19 or its pharmaceutically acceptable salt or solvate or the pharmaceutical composition of claim 20, for use as a medicament for the treatment and / or prevention of diseases mediated by RAS mutation and RAS amplification.

22. Use of the compound of any one of claims 1 - 19 or its pharmaceutically acceptable salt or solvate or the pharmaceutical composition according to claim 20 in the manufacture of a medicament for the prevention or treatment of diseases mediated by RAS mutation and RAS amplification.

23. Use according to claim 22, wherein the diseases mediated by RAS mutation and RAS amplification are selected from: pancreatic cancer, lung cancer, lung adenocarcinoma, bone cancer, skin cancer, head and neck cancer, melanoma of the skin or eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumors (CNS), primary CNS lymphoma, spinal tumors, brainstem glioma or pituitary adenoma.

24. Use according to claim 22, wherein the diseases mediated by RAS mutation and RAS amplification are selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, lung cancer, cholangiocarcinoma, endometrial cancer, ovarian cancer, leukemia; most preferably selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, cholangiocarcinoma.

25. A method for treating and / or preventing a disease mediated by an RAS protein, especially a mutant KRAS protein and KRAS amplification, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1-19 or a pharmaceutically acceptable salt or solvate thereof or a pharmaceutical composition according to claim 20.

26. The method according to claim 25, wherein the diseases mediated by RAS mutation and RAS amplification are selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, lung cancer, cholangiocarcinoma, endometrial cancer, ovarian cancer, leukemia; most preferably selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, cholangiocarcinoma.

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