Kras inhibitors for cancer treatment

Abstract

Compounds having the structure of formula (I) that can be used as KRas inhibitors, pharmaceutical compositions containing such compounds, methods for preparing such compounds, and the use of these compounds in the treatment of cancer.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry. More specifically, this invention relates to a class of compounds with novel structures that can be used as KRas inhibitors, pharmaceutical compositions comprising such compounds, methods for preparing such compounds, and the use of these compounds in the treatment of cancer. Background Technology

[0002] Ras, a homologue of the rat sarcoma oncogene, represents a group of closely related monomeric globular proteins belonging to the GTPase protein family. Specifically, under normal physiological conditions, Ras is activated by growth factors and various other extracellular signals, and is responsible for regulating cell growth, survival, migration, and differentiation. These regulatory functions of Ras are carried out through the switching between GDP-bound and GTP-bound states, or "molecular switches" (Alamgeer et al., Current Opin Pharmacol. 2013, 13:394-401). Ras bound to GDP is an inactive form, in a dormant or closed state, in which the signaling system is shut down. When exposed to certain proliferative stimuli, it is activated, for example, it can be induced by guanine nucleotide exchange factor (GEF) to release GDP and bind to GTP. As a result, Ras is "turned on" and converted into its active form, which recruits and activates various downstream effectors to carry out signal transduction. It can transmit signals from the cell surface to the cytoplasm, thereby controlling many key cellular processes such as differentiation, survival and proliferation (Zhi Tan et al., Mini-Reviews in Medicinal Chemistry, 2016, 16, 345-357).

[0003] Ras possesses GTPase activity, capable of cleaving the terminal phosphate of GTP to convert it into GDP, effectively rendering it inactive. However, Ras' endogenous GTPase activity is very low; the conversion of GTP-Ras to GDP-Ras requires the exogenous protein GAP (GTPase activator). GAP interacts with Ras and promotes the conversion of GTP to GDP. Therefore, any Ras gene mutation affecting the interaction between Ras and GAP or the conversion of GTP to GDP will result in Ras remaining in an activated state for an extended period. This continuously transmits growth and division signals to the cell, stimulating cell proliferation and ultimately leading to tumor formation and development.

[0004] Among human tumor-associated genes, three ubiquitously expressed Ras genes—H-RAS, K-RAS, and N-RAS—encode highly homologous, approximately 21 kDa HRas, NRas, and KRas proteins, respectively. In 1982, researchers first discovered that Ras is activated by mutations in cancer cell lines (Chang, EH et al., Proceedings of the National Academy of Sciences of the United States of America, 1982, 79(16), 4848-4852). Subsequent large-scale genome sequencing studies in different cancer types revealed that Ras proteins are mutated in more than 30% of cancer types, with the highest mutation rates in pancreatic cancer (>90%), colon cancer (45%), and lung cancer (35%). Transgenic and genetically engineered mouse models have also revealed that mutated Ras proteins are sufficient to drive and induce multiple types of cancer, and that Ras oncogenes are crucial for the maintenance and progression of tumors in various cancer types. For example, in Ras-mutated cancer cell lines and animal models, RNA intervention has been shown to slow tumor growth. These studies make Ras tumor proteins a widely accepted and highly attractive target for anticancer drugs 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 Ras mutation-driven cancers. The vast majority of Ras mutations occur at codons G12, G13, and Q61, with approximately 80% of KRas mutations occurring at the glycine residue of codon 12, such as G12C, G12D, and G13D mutations. KRas mutations are commonly found in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and bile duct cancer, and are also observed in 25% of non-small cell lung cancer patients (McCormick, F. et al., Clinical Cancer Research 21(8), 1797-1801, 2015). Therefore, KRas mutant proteins have become the most important branch of Ras drug target research, and the development of its inhibitors is considered a very promising research direction in anticancer / tumor drug development.

[0006] However, drug development targeting Ras over the past three decades has shown that the smooth surface of the Ras protein, lacking obvious grooved or pocket-like structures for binding small molecule inhibitors, and its extremely high affinity for guanine substrates (picomolar level), has made the development of its small molecule inhibitors a difficult problem to solve. As a result, Ras has long been considered an "untreatable" target in the industry. Nevertheless, the continuous efforts to target Ras mutant proteins have yielded some encouraging results. A series of Ras inhibitors have been developed, which inhibit Ras, especially KRas mutations, through multiple pathways, including direct targeting of Ras, inhibition of Ras expression levels, disruption of Ras protein localization, targeted synthesis of lethal components, targeting of Ras-GEF interactions, targeting of Ras-effector interactions, and targeting of Ras dimerization (Zhi Tan et al., Mini-Reviews in Medicinal Chemistry, 2016, 16, 345-357).

[0007] Developed Ras inhibitors, such as those targeting the most common mutant protein KRas-G12C, include allosteric covalent inhibitors, such as the 6H05 series, quinazoline series, ARS series, and tetrahydropyridopyrimidine series, as well as positively bound covalent inhibitors. These inhibitors have been reviewed in the literature (Duan Ni et al., Pharmacology & Therapeutics, https: / / doi.org / 10.1016 / j.pharmthera.2019.06.007). Several patent documents also describe KRas inhibitors of various structural types, such as CN10256421, US2019 / 0144444A1, and WO2019 / 110751A1.

[0008] However, the aforementioned KRas inhibitors still have unresolved issues, and their "drug-likeness" remains unsatisfactory. For example, many KRas-dependent cancers are prone to developing resistance to these targeted therapies, exhibit side effects such as off-target effects, produce chemically active metabolites, have poor metabolic stability, or produce immunogenic covalent adducts (John P. O'Bryan et al., Pharmacological Research 139(2019)503-511; Duan Ni et al., Pharmacology & Therapeutics, https: / / doi.org / 10.1016 / j.pharmthera.2019.06.007). Therefore, there is still a need in clinical practice for more KRas inhibitors with comparable or improved KRas inhibitory activity, improved "drug-likeness," better safety such as fewer drug interactions or metabolic properties, improved pharmacokinetic properties, and / or higher selectivity for different patient populations or specific tumor types.

[0009] This invention provides novel inhibitory compounds with KRas mutant protein inhibitory activity. These compounds, especially the preferred compounds, have the following technical advantages compared to existing KRas mutant protein inhibitors due to their improved structural patterns:

[0010] (1) It retains a considerable or enhanced, or even significantly enhanced, KRas mutant protein and related cancer cell proliferation inhibitory activity;

[0011] (2) Different bioactive spectra are used for different disease types or patient groups;

[0012] (3) It has improved metabolic stability, resulting in better pharmacokinetic properties;

[0013] (4) It has improved physicochemical properties, thus having good drug-like properties and safety, such as being more easily absorbed in the body.

[0014] Invention Summary

[0015] Through research, the inventors have discovered that compounds of formula (I) as defined herein, their isomers, or their pharmaceutically acceptable salts or solvates are effective inhibitors of Ras mutant, especially KRas mutant proteins, capable of inhibiting Ras mutant, especially KRas activity in cells, and can be used to treat or prevent diseases or conditions mediated by or benefiting from the inhibition of Ras mutant, especially KRas mutant proteins, particularly by inhibiting abnormal cell proliferation, thereby treating or preventing tumors or cancers.

[0016] The first aspect of the present invention provides compounds of formula (I), isomers thereof, or pharmaceutically acceptable salts or solvates thereof.

[0017]

[0018] in,

[0019] A is selected from CR a Or N, where R a Selected from halogen, CN, nitro, C 3-6 cycloalkyl or optionally halogenated C 1-6 alkyl;

[0020] R 1 R 2 and R 3 Each is independently selected from H, halogen, or C. 1-6 Alkyl group, wherein the alkyl group is optionally independently selected from halogen, -N(R) c )2、-OR c Or substitution with groups of 3-8 membered heterocyclic alkyl groups;

[0021] R b Each time it appears, it is independently selected from H, halogen, CN, or C that is optionally substituted by halogen or CN. 1-6 alkyl;

[0022] X is selected from the bond, -O-, or -NH-;

[0023] R 4 Selected from H, halogens, C 1-6 Alkyl, -C 0-6 Alkyl-C 6-10 Aryl, -C 0-6 Alkyl-C 3-8 cycloalkyl, -C 0-6 Alkyl-C 3-8 Cycloalkenyl, -C 0-6 Alkyl-3-8-membered heterocyclic alkyl, -C 0-6 alkyl-3-8-membered heterocyclic alkenyl, -C 0-6 Alkyl-5-10-membered heteroaryl, wherein the alkyl, aryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, and heteroaryl groups are optionally substituted by one or more groups independently selected from the following: halogen, C groups optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -SR c 、-(CR c R c ) 0-6 -(CO) 0-1-OR c 、-(CR c R c ) 0-6 -(CO) 0-1 -N(R c )2, wherein -(CR) is attached to an aryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, or heteroaryl group. c R c ) 0-6 -(CO) 0-1 -N(R c )2. Selectively form a 4-7 member nitrogen-containing cyclic group by means of the group attached to N, together with the atoms on the cyclic group to which it is attached and the adjacent atoms;

[0024] R c Each time it appears, it is independently selected from H or C that is optionally substituted with halogen. 1-6 Alkyl groups, or two R atoms attached to the same carbon or nitrogen atom. c Each of them independently forms a 3-6 membered cyclic group together with the carbon or nitrogen atoms to which they are attached;

[0025] E is selected from halogens, -OR d or -N(R) d )2-, where R d Each is independently H or C that is optionally substituted with a halogen. 1-6 Alkyl; R 5 for

[0026] m is 0 or 1;

[0027] Indicates a aromatic ring;

[0028] B and D are each independently selected from N or C;

[0029] Z, G, and Y are each independently selected from C, N, O, or S;

[0030] R 6 R 7 and R 8 Each is independently selected from H, halogens, and C that is optionally substituted with a halogen. 1-6 alkyl;

[0031] The conditions are that B and D are not both N; and at most three of Z, G, Y, D, and B are not C.

[0032] The present invention further provides compounds of formula (II), isomers thereof, pharmaceutically acceptable salts or solvates.

[0033]

[0034] Where R 1 R 2 R 3 R 4 R 6 R 7 R 8 A, R b Each has the meaning given above for the compound of formula (I).

[0035] A second aspect of the invention provides pharmaceutical compositions comprising compounds of formula (I) or formula (II) of the invention, their isomers, or pharmaceutically acceptable salts or solvates thereof.

[0036] The third aspect of the invention provides compounds of formula (I) or formula (II), their isomers, or pharmaceutically acceptable salts or solvates thereof for use as pharmaceuticals.

[0037] The fourth aspect of the invention provides compounds of formula (I) or formula (II), isomers thereof, or pharmaceutically acceptable salts or solvates thereof for the treatment and / or prevention of diseases mediated by Ras mutations, especially KRas mutations, preferably KRasG12C, KRas G12D or KRas G13D mutations, most preferably KRas G12C mutations.

[0038] The fifth aspect of the invention provides the use of compounds of formula (I) or (II), isomers thereof, or pharmaceutically acceptable salts or solvates thereof, or pharmaceutical compositions comprising thereof, in the preparation of medicaments for treating and / or preventing diseases mediated by Ras mutations, especially KRas mutations, preferably KRas G12C, KRas G12D, or KRas G13D mutations, most preferably KRas G12C mutations.

[0039] The sixth aspect of the invention provides a method for treating and / or preventing diseases mediated by Ras mutations, particularly KRas mutations, preferably KRasG12C, KRas G12D or KRas G13D mutations, most preferably KRas G12C mutations, comprising administering to a subject in need a therapeutically effective amount of a compound of formula (I) or formula (II) of the invention, an isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising such a compound.

[0040] The seventh aspect of the present invention provides a method for preparing compounds of formula (I) or formula (II) of the present invention, their isomers, or pharmaceutically acceptable salts or solvates thereof.

[0041] The eighth aspect of the present invention provides a pharmaceutical composition comprising a compound of formula (I) or formula (II) of the present invention, its isomers or pharmaceutically acceptable salts or solvates thereof, and one or more other pharmaceutically active agents. Invention Details

[0043] definition

[0044] Unless otherwise stated, the various terms used in the specification and claims have the meanings shown below. Where a particular term or phrase is not specifically defined, it should not be considered uncertain or unclear, but rather should be understood in accordance with its ordinary meaning in the art. Many of the groups defined herein are optionally substituted, and the list of substituents given in this definition section is merely exemplary and not intended to limit the substituents defined in other parts of this specification and claims.

[0045] As used herein, the term "Ras mutation" or "Ras mutant protein" refers to a protein encoded and expressed by the Ras gene with mutations in one or more of its codons. This typically includes, but is not limited to, Ras proteins with mutations in the glycine residue at codon 12, codon 13, or glutamine residue at codon 61, such as mutant HRas, NRas, or KRas. These residues are located at the active site of Ras, and mutations in these residues can impair Ras's inherent or GAP-catalyzed GTPase activity, resulting in the persistence of GTP-bound Ras.

[0046] For the purposes of this invention, "Ras mutation" or "Ras mutant protein" may be used interchangeably, and generally refers to mutated HRas, NRas, or KRas, such as, but not limited to, HRas-G12C (a mutation from glycine to cysteine ​​at codon G12), NRas-G12C, KRas-G12C, KRas-G12D (a mutation from glycine to aspartic acid at codon G12), and KRas-G13D (a mutation from glycine to aspartic acid at codon G13); particularly refers to KRas mutant protein, more particularly refers to KRas-G12C mutant protein, KRas-G12D mutant protein, KRas-G13D mutant protein, and most particularly refers to KRas-G12C mutant protein.

[0047] As used herein, the term "treatment" means administering one or more compounds of formula (I) described herein, their isomers, or pharmaceutically acceptable salts or solvates thereof to a subject, such as a mammal or a human, who suffers from or has symptoms of the disease, in order to cure, alleviate, reduce, or affect the disease or its symptoms. In a specific embodiment of the invention, the disease is a Ras mutation-mediated disease as defined below, particularly a tumor or cancer.

[0048] As used herein, the term "prevention" is well-known in the art and refers to the administration of one or more compounds of formula (I) described herein, isomers thereof, or pharmaceutically acceptable salts or solvates thereof to a subject, such as a mammal or human, suspected of having or susceptible to a Ras mutation-mediated disease, particularly cancer or tumor, as defined herein, thereby reducing the risk of developing the defined disease. The term "prevention" also includes the use of compounds of the present invention prior to the diagnosis or determination of any clinical and / or pathological symptoms.

[0049] As used herein, the terms “inhibition” and “reduction”, or any variations thereof, refer to the ability of a bioactive agent to reduce the signal transduction activity of a target by interacting directly or indirectly with the target, and to any measurable reduction or complete inhibition of the target activity. For example, this reduction in activity (e.g., KRas activity) compared to normal conditions can be 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 derived therefrom.

[0050] As used herein, the term "selective inhibition" refers to the ability of a bioactive agent to preferentially reduce the signal transduction activity of a target site, compared to off-target signaling activity, through direct or indirect interaction with the target site. With respect to the compound of formula (I) of the invention, it has the ability to selectively inhibit G12 or G13 mutations in KRas, HRas, or NRas proteins, such as G12C, G12D, and G13D mutations, with respect to various mutations occurring in one or more codons of the Ras protein, preferably selectively inhibiting G12C mutations in the KRas protein. For example, compared to another specific Ras mutation, the present invention has a better inhibitory activity against a specific Ras mutation by at least 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 thereof, or has a better activity against a specific Ras mutation (e.g., KRas-G12C) by at least 1-, 2-, 3-, 4-, 5-, 10-, 25-, 50-, 100-, 250- or 500-fold compared to the activity against another specific Ras mutation, or has a better activity against a specific Ras mutation (e.g., KRas-G12C) by at least 1-, 2-, 3-, 4-, 5-, 10-, 25-, 50-, 100-, 250- or 500-fold.

[0051] As used herein, the term "Ras mutation-mediated disease" refers to a disease in which Ras mutations promote the occurrence and development of the disease, or in which inhibiting Ras mutations reduces the incidence of the disease, decreases or eliminates the symptoms. For the purposes of this invention, "Ras mutation-mediated disease" preferably refers to a disease mediated by KRas mutations, most preferably a disease mediated by KRas-G12C, and even more preferably cancer or tumors.

[0052] As used herein, the terms “cancer” or “tumor” refer to abnormal cell growth and proliferation, whether malignant or benign, and all precancerous cells and cancerous tissues. For all aspects of this invention, the cancers or tumors include, but are 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, anal region cancer, gastric cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small bowel 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 ureter cancer, renal cell carcinoma, renal pelvis cancer, central nervous system tumors (CNS), primary CNS lymphoma, spinal tumors, brainstem gliomas, or pituitary adenomas.

[0053] In all respects of the invention, preferably, the cancer or tumor is associated with Ras mutations, particularly KRas mutations, preferably KRas G12C, KRas G12D, or KRas G13D mutations, and most preferably KRas G12C mutations, including but not limited to the aforementioned tumor types and their preferred ranges. Particularly preferred tumors of the invention include lung cancer, lung adenocarcinoma, colon cancer, rectal cancer, pancreatic cancer, endometrial cancer, cholangiocarcinoma, leukemia, and ovarian cancer.

[0054] As used herein, the terms “subject,” “individual,” or “patient” refer to a vertebrate. In some embodiments, the vertebrate is a mammal. Mammals include, but are not limited to, farm animals (such as cattle), sport animals, pets (such as guinea pigs, cats, dogs, rabbits, and horses), primates, mice, and rats. In some embodiments, the mammal is a human.

[0055] As used herein, the term "therapeutic effective amount" refers to an amount or dose that is generally sufficient to produce a beneficial therapeutic effect for patients with cancer or tumors requiring treatment. Those skilled in the art can determine the effective amount or dose of the active ingredient in this invention using conventional methods, in conjunction with common influencing factors.

[0056] As used herein, the term "drug combination" refers to the combination of the compounds of the present invention with other active agents to achieve the objectives of the present invention. These other active agents may be one or more other compounds of the present invention, or may be a second or additional (e.g., a third) compound that is compatible with the compounds of the present invention, i.e., does not adversely affect each other, or has complementary activity. For example, these active agents are known to regulate other biological pathways, or regulate different components of the biological 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 intended purpose. These other active agents may be co-administered with the compounds of the present invention in a single pharmaceutical composition, or may be administered separately from the compounds of the present invention in different discrete units, and when administered separately, they may be administered simultaneously or sequentially. The sequential administration may be close in time or spaced apart.

[0057] On the one hand, 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, mitosis inhibitors, topoisomerase inhibitors, anti-hormonal drugs, angiogenesis inhibitors or cytotoxic agents.

[0058] As used in this article, the term "pharmaceutically acceptable" means a molecular entity or composition that, when administered in appropriate amounts to animals such as humans, will not produce adverse, allergic, or other adverse reactions.

[0059] As used in this article, the term "pharmaceutically acceptable salt" refers to salts that retain the biological efficacy and properties of the parent compound and are not biologically or otherwise undesirable, including acid addition salts and base addition salts. "Pharmaceutically acceptable acid addition salts" can be formed by compounds with a free base and inorganic or organic acids. Inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, etc., while organic acids can be selected from aliphatic, alicyclic, aromatic, arylaliphatic, heterocyclic, carboxylic acid, and sulfonic acid 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, dihydroxynaphthyl 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, and aluminum, as well as salts derived from pharmaceutically acceptable non-toxic organic bases, including but not limited to primary, secondary, and tertiary amines, substituted ammonium compounds, 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, heparin, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, triethanolamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc.

[0060] As used herein, the term "isomer" refers to any stereoisomer, enantiomer mixture, including racemic mixtures, diastereomer mixtures, geometric isomers, blocked isomers, and / or tautomers that may exist structurally as a compound. The methods for determining and isolating the stereochemistry of such isomers are well known to those skilled in the art (SP 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). Therefore, this invention covers all possible isomer forms of the compound of formula (I) as defined above, and their pharmaceutically acceptable salts or solvates.

[0061] The compound structural formulas or structural fragments used in this article The absolute configuration of the chiral center is indicated by R and S in the nomenclature of the compounds or intermediates provided in this invention. The determination of the absolute configuration is well known to those skilled in the art.

[0062] The structural fragments discussed in this article use The bonds that cross it are the bonds that connect the structural segment to the rest of the molecule.

[0063] As used herein, the term "solvent" refers to a solvation form containing a stoichiometric or non-stoichiometric 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 methanolides, ethanolides, or acetonitrileides, respectively; or in any polymorphic form. It should be understood that such solvates of the compounds of the present invention also include solvates of pharmaceutically acceptable salts of the compounds of the present invention.

[0064] As used herein, the term "isotope variant" refers to a compound in which one or more atoms constituting the compound contain an isotope in a non-natural proportion. The compounds of the present invention may contain an atomic isotope in a non-natural proportion on one or more atoms constituting the compound, thereby forming isotopic variations of the compounds of the present invention or pharmaceutically acceptable salts thereof, whether or not they are radioactive, and are intended to be covered within the scope of the present invention. Examples of isotopes that may be incorporated into the compounds of the present invention and pharmaceutically acceptable salts thereof include, for example... 2 H, 3 H, 13 C 14 C 15 N、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F and 36 Cl. It should be understood that isotopic variations of the compounds of the present invention and their pharmaceutically acceptable salts can generally be prepared by conventional methods using appropriate isotopic variations with suitable reagents. For example, radioactive isotopes may be incorporated (e.g., 3 H or 14 C) Certain isotopic variations of the compounds of this invention and their pharmaceutically acceptable salts may be used for drug and / or substrate tissue distribution studies. Tritium substitution... 3 H and carbon-14, i.e. 14 Carbon isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, isotopes such as deuterium are also used. 2H substitution can provide certain therapeutic advantages due to increased metabolic stability, such as increased in vivo half-life or reduced dose requirements, and is therefore preferred in some cases. Additionally, it is possible to prepare positron-emitting isotopes (e.g., 11 C 18 F, 15 O and 13 The compounds of the present invention, which are substituted with N), can be used in positron emission tomography (PET) studies for the detection of substrate receptor occupancy.

[0065] As used herein, the term "metabolite" refers to a product generated from the metabolism of a particular compound in the body. Such products may arise, for example, from the oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, etc., of the applied compound. The identification and analysis of metabolite products are performed in a manner well known to those skilled in the art.

[0066] As used herein, the term "prodrug" refers to a compound that can be converted into a bioactive compound, such as a compound of formula (I), under physiological conditions or by solvent degradation. Therefore, the term "prodrug" refers to a pharmaceutically acceptable precursor of a bioactive compound. In some respects, a prodrug is inactive when administered to a subject, but is converted into an active compound in vivo, for example, by hydrolysis. Prodrug compounds often offer advantages in mammalian organisms such as solubility, tissue compatibility, or delayed release (see, for example, Bundgard, H., Design of Prodrugs (1985), pp. 7–9, 21–24 (Elsevier, Amsterdam). Discussions of prodrugs can be found in Higuchi, T. et al., ACS Symposium Series, Vol. 14, and "Bioreversible Carriers in Drug Design," Edward B. Roche, Pharmaceutical Association & Pergamon. Press, 1987, the entire contents of which are incorporated herein by reference. The term “prodrug” also means any covalently bonded carrier that, when administered to a mammalian subject, releases the active compound in vivo. Prodrugs of the active compounds described herein are typically prepared by modifying functional groups present in the active compound such that the modification can be cleaved into the parent active compound in routine operation or in vivo. Prodrugs include compounds in which a hydroxyl, amino, or thiol group is bonded to a hydroxyl, amino, or thiol group that cleaves upon administration of the prodrug to a mammal to form a free hydroxyl, free amino, or free thiol group. Any functional group. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives with hydroxyl functional groups, or acetamide, formamide, and benzamide derivatives with amine functional groups in the active compound. In some embodiments, prodrugs include phosphate / ester-containing prodrugs, borate-containing prodrugs, thiophosphate / ester-containing prodrugs, sulfate / ester-containing prodrugs, peptide-containing prodrugs, D-amino acid-modified prodrugs, glycosylated prodrugs, β-lactam-containing prodrugs, prodrugs containing optionally substituted phenoxyacetamides or optionally substituted phenylacetamides, as well as 5-fluorocytosine and 5-fluorouridine prodrugs.

[0067] As used herein, the term "pharmaceutically acceptable excipient or carrier" refers to one or more compatible solid or liquid fillers or gelling substances suitable for human use and having sufficient purity and sufficiently low toxicity. Examples include, but are not limited to, cellulose and its derivatives (such as sodium carboxymethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as magnesium stearate), calcium sulfate, vegetable oils, polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween compounds), wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, etc.

[0068] As used herein, the terms “halogen” or “halogenated” refer to F, Cl, Br, or I. Furthermore, the term “halogen-substituted” is intended to include monohalogenated or polyhalogenated groups, wherein one or more hydrogen atoms are present in one or more identical or different halogen-substituted groups.

[0069] As used herein, the term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group consisting of carbon and hydrogen atoms. Specifically, an alkyl group has 1 to 10, for example, 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 "C1-C6 alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms, examples of which include methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, sec-butyl, or tert-butyl), pentyl (including n-pentyl, isopentyl, and neopentyl), n-hexyl, 2-methylpentyl, etc. Certain alkyl groups have 1 to 3 carbon atoms.

[0070] As used herein, the term "alkoxy" refers to the group -O-alkyl, where alkyl has the meaning as described herein. Specifically, the term includes the group -OC. 1-6 Alkyl, more specifically -OC 1-3 Alkyl groups. Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, and tert-butoxy), pentoxy (including n-pentoxy, isopentoxy, and neopentoxy), and hexoxy (including n-hexoxy and isohexoxy). Specific alkoxy groups have 1 to 3 carbon atoms.

[0071] As used herein, the term "alkylthio" refers to -S-alkyl, where the alkyl group is as defined above for "alkyl". Specifically, the term includes the -SC group. 1-6 Alkyl, more specifically -SC 1-3 Alkyl groups. Representative examples of alkylthio groups include, but are not limited to, methylthio, ethylthio, propylthio (including n-propylthio and isopropylthio), butylthio (including n-butylthio, isobutylthio, and tert-butylthio), pentylthio (including n-pentylthio, isopentylthio, and neopentylthio), and hexylthio (including n-hexylthio and isohexylthio). Specific alkylthio groups have 1 to 3 carbon atoms.

[0072] As used herein, the term "halogen-substituted C1-C6 alkyl" refers to the C1-C6 alkyl group described above, wherein one or more (e.g., 1, 2, 3, 4, or 5) hydrogen atoms are replaced by a halogen. Those skilled in the art will understand that when there is more than one halogen substituent, the halogens can be the same or different, and can be located on the same or different C atoms. Examples of "halogen-substituted C1-C6 alkyl" include, for example, -CH2F, -CHF2, -CF3, -CCl3, -C2F5, -C2Cl5, -CH2CF3, -CH2Cl, -CH2CH2CF3, or -CF(CF3)2, etc.

[0073] As used herein, the term "cycloalkyl" refers to a monocyclic, fused polycyclic, bridged polycyclic, or spirocyclic non-aromatic saturated monovalent hydrocarbon ring structure having a specified number of ring atoms. Cycloalkyl groups can have 3 to 12 carbon atoms (i.e., C3-C4). 12 Cycloalkyl groups, for example, with 3 to 10, 3 to 8, 3 to 7, 3 to 6, or 5 to 6 carbon atoms. Examples of suitable cycloalkyl groups include, but are not limited to, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl; or polycyclic (e.g., bicyclic) structures, including spirocyclic, fused, or bridged systems such as bicyclic [1.1.1]pentyl, bicyclic [2.2.1]heptyl, spiro[3.4]octyl, bicyclic [3.1.1]hexyl, bicyclic [3.1.1]heptyl, or bicyclic [3.2.1]octyl, etc.

[0074] As used herein, the term "cycloalkenyl" refers to a monocyclic, fused polycyclic, bridged polycyclic, or spirocyclic nonaromatic unsaturated hydrocarbon ring structure having a specified number of ring atoms and containing at least one (e.g., 1, 2, or 3) carbon-carbon double bonds. Cycloalkenyl groups can have 3 to 12 carbon atoms (i.e., C3-C4). 12 Cycloalkenyl groups, for example, with 3 to 10, 3 to 8, 3 to 7, 3 to 6, or 5 to 6 carbon atoms. Suitable examples of cycloalkenyl groups include, but are not limited to, monocyclic structures such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, cycloheptanetrienyl, or cyclooctenyl.

[0075] As used herein, the term "heterocyclic alkyl" means a monocyclic, fused polycyclic, spirocyclic, or bridged polycyclic nonaromatic saturated ring structure comprising one or more (e.g., 1, 2, 3, or 4) heteroatoms independently selected from O, N, and S, and a specified number of ring atoms, or its N-oxide, or its S-oxide or S-dioxide. Heterocyclic alkyls may have 3 to 12 ring members (which may be referred to as 3-12 membered heterocyclic alkyls), for example, 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, or 5 to 6 ring members. Heterocyclic alkyls typically contain up to 4 (e.g., 1, 2, 3, or 4) heteroatoms. Suitable examples of heterocyclic alkyl groups include, but are not limited to, azaheterocyclic butyl, oxacyclobutyl, thioheterocyclic butyl, pyrrolidinyl (e.g., 1-pyrrolidinyl, 2-pyrrolidinyl, and 3-pyrrolidinyl), tetrahydrofuranyl (e.g., 1-tetrahydrofuranyl, 2-tetrahydrofuranyl, and 3-tetrahydrofuranyl), tetrahydrothiophenyl (e.g., 1-tetrahydrothiophenyl, 2-tetrahydrothiophenyl, and 3-tetrahydrothiophenyl), and piperidinyl (e.g., 1-piperidinyl). 2-piperidinyl, 3-piperidinyl, and 4-piperidinyl), tetrahydropyranyl (e.g., 4-tetrahydropyranyl), tetrahydrothiaranyl (e.g., 4-tetrahydrothiaranyl), morpholinyl (e.g., morpholino), thiomorpholinyl, dioxane, piperazine, or aziridine heptyl, diazacyclic heptyl such as 1,4-diazacyclic heptyl, 3,6-diaza-bicyclo[3.1.1]heptyl, or 3-aza-bicyclo[3.2.1]octyl. The atom in the heterocyclic alkyl group attached to the rest of the compound can be a carbon atom or a heteroatom, as long as it is chemically feasible.

[0076] Preferred heterocyclic alkyl groups, for example It should be understood that structures with asymmetric centers encompass their racemic and / or singular enantiomeric forms, such as... Can represent and / or

[0077] As used herein, the term "heterocyclic alkenyl" means a "heterocyclic alkyl" as defined herein that contains at least one (e.g., 1, 2, or 3) double bonds. Examples of suitable heterocyclic alkenyl groups include, but are not limited to:

[0078]

[0079] Each W is selected from CH2, NH, O, and S; each Y is selected from NH, O, C (=O), SO2, and S; and each Z is selected from N and CH, provided that each ring contains at least one atom selected from N, O, or S; for example, pyrrolinyl (e.g., 1-pyrrolinyl, 2-pyrrolylalkyl, 3-pyrrolinyl, 4-pyrrolinyl, or 5-pyrrolinyl), dihydrofuranyl (e.g., 1-dihydrofuranyl, 2-dihydrofuranyl, 3-dihydrofuranyl, 4-dihydrofuranyl, or 5-dihydrofuranyl), dihydrothiophenyl (e.g., 1-dihydrothiophenyl, 2-dihydrothiophenyl, 3-dihydrothiophenyl, or 4-dihydrothiophenyl), tetrahydropyridyl (e.g., 1-, 2-, 3-, 4-, 5-, or 6-tetrahydropyridyl), tetrahydropyranyl (e.g., 4-tetrahydropyranyl), or tetrahydrothiophenyl (e.g., 4-tetrahydrothiophenyl).

[0080] As used herein, the term "aryl" refers to a monovalent aromatic hydrocarbon group derived by removing a hydrogen atom from a single carbon atom in an aromatic ring system. Specifically, aryl refers to a monocyclic or fused polycyclic aromatic ring structure having a specified number of ring atoms. More specifically, the term includes groups comprising 6 to 14, for example 6 to 10, preferably 6 ring members. Specific aryl groups include phenyl and naphthyl, with phenyl being the most specific.

[0081] As used herein, the term "heteroaryl" refers to a monocyclic or fused polycyclic aromatic ring structure comprising one or more (e.g., 1, 2, 3, or 4) heteroatoms independently selected from O, N, and S, and a specified number of ring atoms, or its N-oxide, or its S-oxide or S-dioxide. Specifically, the aromatic ring structure may have 5 to 10 ring members. The heteroaryl may be, for example, a 5-6 membered monocyclic ring, or a fused bicyclic structure formed by two fused 6-membered rings, two fused 5-membered rings, a fused 6-membered ring and a fused 5-membered ring, or a fused 5-membered ring and a 4-membered ring. The heteroaryl ring will typically contain up to 4 heteroatoms, more typically up to 3 heteroatoms, and more typically up to 2 heteroatoms, for example, a single heteroatom independently selected from O, N, and S, wherein N and S may be in oxidized states such as N oxide, S=O, or S(O)2. In one embodiment, the heteroaryl ring contains at least one cyclic nitrogen atom, at least one cyclic sulfur atom, or at least one epoxy atom. For example, the heteroaryl group can be a fused ring containing 1, 2, 3, or 4 heteroatoms independently selected from N, O, or S, such as benzofuran, benzothiophene, indole, benzimidazole, indazole, benzotriazole, pyrrolo[2,3-b]pyridine, pyrrolo[2,3-c]pyridine, pyrrolo[3,2-c]pyridine, pyrrolo[3,2-b]pyridine, imidazo[4,5-b]pyridine. Pyridine, imidazo[4,5-c]pyridine, pyazolo[4,3-d]pyridine, pyazolo[4,3-c]pyridine, pyazolo[3,4-c]pyridine, pyazolo[3,4-b]pyridine, isoindole, purine, indene, imidazo[1,2-a]pyridine, imidazo[1,5-a]pyridine, pyazolo[1,5-a]pyridazine, pyrrolo[1,2-b]pyrimidine Imidazolo[1,2-c]pyrimidine, 5H-pyrrolo[3,2-b]pyrazine, 1H-pyrazolo[4,3-b]pyrazine, 1H-pyrazolo[3,4-d]pyrimidine, 7H-pyrrolo[2,3-d]pyrimidine, quinoline, isoquinoline, cinnamoline, quinazoline, quinoxaline, phthalazine, 1,6-naphthidine, 1,7-naphthidine, 1,8-naphthidine, 1,5-naphthidine, 2, 6-Naphthyl, 2,7-Naphthyl, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-d]pyrimidine, pyrido[2,3-d]pyrimidine, pyrido[2,3-b]pyrazine, pyrido[3,4-b]pyrazine, pyrimido[5,4-d]pyrimidine, pyrazido[2,3-b]pyrazine, and pyrimido[4,5-d]pyrimidine. For example, the heteroaryl group can be a 5- or 6-membered heteroaryl group containing one or two heteroatoms independently selected from N, O, or S. Examples of suitable 5-membered monocyclic heteroaryl groups include, but are not limited to, pyrrole, furanyl, thiophene, imidazolyl, furazonyl, oxazolyl, oxadiazolyl, oxtriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl, and tetrazolyl; examples of suitable 6-membered monocyclic heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyridinyl, pyrimidinyl, and triazinyl.The atoms that are attached to the rest of the compound in a heteroaryl group can be carbon atoms or heteroatoms, as long as it is chemically feasible.

[0082] The description of a substituent as "optionally substituted" means that the group can be unsubstituted or substituted by one or more (e.g., 0, 1, 2, 3, 4, or 5 or more, or any range thereof) of the listed substituents, wherein the substituents can be the same or different. In one embodiment, the optionally substituted group is substituted by 1 substituent. In another embodiment, the optionally substituted group is substituted by 2 substituents. In another embodiment, the optionally substituted group is substituted by 3 substituents. In yet another embodiment, the optionally substituted group is substituted by 4 substituents.

[0083] As is understood by those skilled in the art of organic synthesis, the maximum number of heteroatoms or the type of heteroatoms in a stable, chemically viable heterocycle, whether aromatic or non-aromatic, is determined by the ring size, degree of unsaturation, and valence of the heteroatoms. Generally, a heterocycle can have 1 to 4 heteroatoms, provided that the heterocycle or heteroaromatic ring is chemically viable and stable.

[0084] The term "hydroxyl group" as used in this article refers to the -OH group.

[0085] The term "thiol" as used in this article refers to the -SH group.

[0086] The term "nitro" as used in this article refers to the -NO2 group.

[0087] As used herein, the term "optionally substituted" means, unless otherwise indicated, that a group may be unsubstituted or substituted by one or more (e.g., 0, 1, 2, 3, 4, or 5 or more, or any range thereof) of the substituents listed for that group, wherein said substituents 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 substituents. In another embodiment, the optionally substituted group has 3 substituents. In another embodiment, the optionally substituted group has 4 substituents. In another embodiment, the optionally substituted group has 5 substituents.

[0088] Unless otherwise specified, C in the definition of compounds in this invention n-n+m Or C n -C m This includes various cases with n to n+m carbons, such as C 1-6 This includes C1, C2, C3, C4, C5, and C6, as well as any range from n to n+m, such as C 0-6 Including C1, C2, C3, C4, C5, C6, C 0-1 C 0-2C 0-3 C 0-4 C 0-5 C 1-2 C 1-3 C 1-4 C 2-3 C 1-6 Including C 1-2 C 1-3 C 1-4 C 2-6 C 3-6 Similarly, in the compound definition of this invention, n- to n+m-membered rings represent the number of ring atoms from n to n+m. For example, 3-12-membered rings include 3-membered rings, 4-membered rings, 5-membered rings, 6-membered rings, 12-membered rings, etc., and also include any range from n to n+m-membered rings. For example, 3-12-membered rings include 3-6-membered rings, 3-8-membered rings, 3-9-membered rings, 4-7-membered rings, 4-5-membered rings, 5-6-membered rings, 5-7-membered rings, 5-8-membered rings, 5-9-membered rings, 6-7-membered rings, 6-8-membered rings, and 6-10-membered rings, etc.

[0089] As used in this specification and the following claims, the word “comprising” and variations thereof 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 comprising multiple ingredients, steps, or conditions, it should be understood that the element may also be described as comprising any combination of said multiple ingredients, steps, or conditions, or “consisting of multiple or combined ingredients, steps, or conditions” or “consisting substantially of multiple or combined ingredients, steps, or conditions.”

[0090] It should be understood that the dosages referred to herein when describing the compounds of the present invention, pharmaceutical compositions comprising them, pharmaceutical combinations, cassettes, and related uses and methods are based on weight in free form and do not include any salts, hydrates, or solvates thereof, unless otherwise specified in the specification that the dosage is based on the weight of salts, hydrates, or solvates.

[0091] Compounds of the present invention

[0092] The terms “compound of the invention” and “compound of the invention” as used throughout this application, unless otherwise indicated, encompass compounds of formula (I) or formula (II) as defined in the various embodiments and preferred embodiments herein, including their isomers, including transisomers, enantiomer mixtures, particularly racemates, diastereomer mixtures, geometric isomers, tautomers, solvates, metabolites, isotopic variants, salts (e.g., pharmaceutically acceptable salts) and prodrugs.

[0093] Therefore, all the aforementioned isomers and derivatives of Formula I are covered within the scope of this invention, and their respective meanings, preparation methods, and specific examples are as defined in the "Definitions" section above, or are well known in the art. In some embodiments, metabolites, isotopic variants, or prodrugs, and any combinations thereof, are excluded as appropriate. However, substantially pure enantiomers (enantiomerically pure) or diastereomers of Formula (I) or Formula (II) compounds and / or their pharmaceutically acceptable salts are preferred.

[0094] This invention also covers N-oxides of compounds of formula (I) or (II), provided that these compounds contain a basic nitrogen atom, such as a nitrogen atom present in a nitrogen-containing heterocycle. Some compounds of this invention may exist in polymorphic or amorphous forms, and therefore also fall within the scope of this invention.

[0095] On the one hand, the present invention provides compounds of formula (I), isomers thereof, or pharmaceutically acceptable salts or solvates thereof.

[0096]

[0097] in,

[0098] A is selected from CR a Or N, where R a Selected from halogen, CN, nitro, C 3-6 cycloalkyl or optionally halogenated C 1-6 alkyl;

[0099] R 1 R 2 and R 3 Each is independently selected from H, halogen, or C. 1-6 Alkyl group, wherein the alkyl group is optionally independently selected from halogen, -N(R) c )2、-OR c Or substitution with groups of 3-8 membered heterocyclic alkyl groups;

[0100] R b Each time it appears, it is independently selected from H, halogen, CN, or C that is optionally substituted by halogen or CN. 1-6 alkyl;

[0101] X is selected from the bond, -O-, or -NH-;

[0102] R 4 Selected from H, halogens, C 1-6 Alkyl, -C 0-6 Alkyl-C 6-10 Aryl, -C 0-6 Alkyl-C 3-8 cycloalkyl, -C 0-6 Alkyl-C 3-8 Cycloalkenyl, -C0-6 Alkyl-3-8-membered heterocyclic alkyl, -C 0-6 alkyl-3-8-membered heterocyclic alkenyl, -C 0-6 Alkyl-5-10-membered heteroaryl, wherein the alkyl, aryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, and heteroaryl groups are optionally substituted by one or more groups independently selected from the following: halogen, C groups optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -SR c 、-(CR c R c ) 0-6 -(CO) 0-1 -OR c 、-(CR c R c ) 0-6 -(CO) 0-1 -N(R c )2, wherein -(CR) is attached to an aryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, or heteroaryl group. c R c ) 0-6 -(CO) 0-1 -N(R c )2. Selectively form a 4-7 member nitrogen-containing heterocycle by means of the group attached to the N, together with the atoms on the cyclic group attached thereto and the adjacent atoms;

[0103] R c Each time it appears, it is independently selected from H or C that is optionally substituted with halogen. 1-6 Alkyl groups, or two R atoms attached to the same carbon or nitrogen atom. c Each of them independently forms a 3-6 membered cyclic group together with the carbon or nitrogen atoms to which they are attached;

[0104] E is selected from halogens, -OR d or -N(R) d )2-, where R d Each is independently H or C that is optionally substituted with a halogen. 1-6 Alkyl; R 5 for

[0105] m is 0 or 1;

[0106] Indicates a aromatic ring;

[0107] B and D are each independently selected from N or C;

[0108] Z, G, and Y are each independently selected from C, N, O, or S;

[0109] R 6 R 7 and R 8 Each is independently selected from H, halogens, and C that is optionally substituted with a halogen. 1-6 alkyl;

[0110] The conditions are: B and D are not both N; and at most three of Z, G, Y, D, and B are not C.

[0111] Or its isomers, pharmaceutically acceptable salts or solvates.

[0112] In one embodiment of a compound of formula (I), A is N.

[0113] In one embodiment of a compound of formula (I), A is CR a , where R a Selected from halogens.

[0114] In one embodiment of a compound of formula (I), A is CR a , where R a It can be F or Cl, with Cl being preferred.

[0115] In one embodiment of a compound of formula (I), R 1 Selected from H or halogens.

[0116] In one embodiment of a compound of formula (I), R 1 It can be H or F.

[0117] In one embodiment of a compound of formula (I), R 2 and R 3 All are H.

[0118] In one embodiment of a compound of formula (I), R b For H.

[0119] In one embodiment of a compound of formula (I), R b It is not H, and there exists 1 R. b Or there exist 2 R b .

[0120] In one embodiment of a compound of formula (I), there is an R b And R b C 1-6 alkyl.

[0121] In a specific implementation, there exists an R b And R b C 1-6 Alkyl, preferably C 1-3Alkyl group; in a more specific embodiment, an R is present. b And R b For -CH3; in a more specific implementation, there exists an R b And R b for In a more specific implementation, there exists an R b And R b for And the connection method on the piperazine ring is as follows In a more specific implementation, there exists an R b And R b for And the connection method on the piperazine ring is as follows

[0122] In one embodiment of a compound of formula (I), there is an R b And R b For CN; in a specific implementation, there exists an R b And R b It is CN and is attached to the carbon atom adjacent to the amide nitrogen.

[0123] In one embodiment of a compound of formula (I), there is an R b And R b C 1-6 Alkyl, preferably C 1-3 Alkyl group, substituted with CN; in specific embodiments, an R is present. b And R b It is cyanomethyl; in a more specific embodiment, there is an R b And R b It is a cyanomethyl group, and the connection mode on the piperazine ring is as follows:

[0124] In one embodiment of a compound of formula (I), two R are present. b And R b C 1-6 Alkyl, preferably C 1-3 Alkyl group; in specific embodiments, two R groups are present. b And R b For -CH3; in a more specific implementation, there are two R... b And R b It is -CH3, and the connection mode on the piperazine ring is as follows:

[0125] In one embodiment of a compound of formula (I), X is a bond.

[0126] In one embodiment of a compound of formula (I), X is -O- or -NH-.

[0127] In one embodiment of a compound of formula (I), X is -O-.

[0128] In one embodiment of a compound of formula (I), X is -NH-.

[0129] In one embodiment of a compound of formula (I), R 4 Selected from H, halogens, C 1-6 Alkyl, -C 0-3 Alkyl-phenyl, -C 0-3 Alkyl-C 3-6 cycloalkyl, -C 0-3 Alkyl-C 3-6 Cycloalkenyl, -C 0-3 Alkyl group - a 4-6 membered heterocyclic alkyl group containing 1, 2 or 3 heteroatoms independently selected from N, O or S, -C 0-3 Alkyl group – comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, consisting of a 5- or 6-membered heterocyclic alkenyl group and a -C group. 0-3 Alkyl group – comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the alkyl, phenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, and heteroaryl groups are optionally substituted by 1, 2, or 3 groups independently selected from: halogen, C group optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -SR c 、-(CR c R c ) 0-6 -OR c 、-(CR c R c ) 0-6 -N(R c )2, wherein -(CR) is attached to a phenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, or heteroaryl group. c R c ) 0-6 -N(R c )2. Selectively form a 4-7 member nitrogen-containing heterocycle by means of the group attached to the N, together with the atoms on the cyclic group attached thereto and the adjacent atoms.

[0130] In one embodiment of a compound of formula (I), R 4 Selected from H, halogens, C 1-6 Alkyl, -C 0-3 Alkyl-phenyl, -C 0-3 Alkyl-C 3-6 cycloalkyl, -C 0-3Alkyl group – a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S and -C 0-3 Alkyl group – comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the alkyl, phenyl, cycloalkyl, heterocycloalkyl, and heteroaryl group is optionally substituted by 1, 2, or 3 groups independently selected from: halogen, C group optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -SR c 、-(CR c R c ) 0-6 -OR c 、-(CR c R c ) 0-6 -N(R c )2, wherein -(CR) is attached to a phenyl, cycloalkyl, heterocycloalkyl, or heteroaryl group. c R c ) 0-6 -N(R c )2. Selectively form a 4-7 member nitrogen-containing heterocycle by means of the group attached to the N, together with the atoms on the cyclic group attached thereto and the adjacent atoms.

[0131] In one embodiment of a compound of formula (I), R 4 Selected from H, halogens, C 1-6 Alkyl, phenyl, C 3-6 Cycloalkyl, 4-6-membered heterocycloalkyl comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, and 5-6-membered heteroaryl comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the alkyl, phenyl, cycloalkyl, heterocycloalkyl, and heteroaryl groups are optionally substituted by 1, 2, or 3 groups independently selected from: halogen, C groups optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -OR c 、-(CR c R c ) 0-6 -N(R c )2, wherein -(CR) is attached to a phenyl, cycloalkyl, heterocycloalkyl, or heteroaryl group. c R c ) 0-6 -N(R c )2. Selectively form a 4-7 member nitrogen-containing heterocycle by means of the group attached to the N, together with the atoms on the cyclic group attached thereto and the adjacent atoms.

[0132] In one embodiment of a compound of formula (I), -XR 4 For H.

[0133] In one embodiment of a compound of formula (I), -XR 4 Not H.

[0134] In one embodiment of a compound of formula (I), -XR 4 It is -OH.

[0135] In one embodiment of a compound of formula (I), -XR 4 It is -NH2.

[0136] In one embodiment of a compound of formula (I), -XR 4 It is a halogen; in the specific implementation scheme, -XR 4 It is Cl or F.

[0137] In one embodiment of a compound of formula (I), R 4 C 1-6 Alkyl groups, optionally selected by one, two, or three independently chosen from halogens, C 1-6 Alkyl, hydroxyl, C 1-6 Alkoxy and -(CR c R c ) 0-6 -N(R c The group substituted for )2. In a specific embodiment, exemplary R 4Including but not limited to -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2CH(CH3)CH3, -CH2CH2CH2CH3, -C(CH3)(CH3)(CH3), -CH2F , -CHF2, -CF3, -CH2CF3, -CH2CH2CF3, -CH(CF3)CH3, -CH2CH(CF3)CH3, -CH2CH2CH2CF3, -C(CF3)(CH3)(CH3), -CH2O H, -CH2CH2OH, -CH2CH2CH2OH, -CH2CH(OH)CH3, -CH2CH(CH2OH)CH3, -CH2CH2CH2CH2OH, -C(CH2OH)(CH3)(CH3), -CH 2OCH3, -CH2OCH2CH3, -CH2CH2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2OCH3, -CH2CH(OCH3)CH3, -CH2CH(CH2OCH3)CH3, -CH2CH2CH2CH2OCH3, -NH2, -NHCH3, -NHCH2CH3, -N(CH3)2, -N(CH3)(CH2CH3), -N(CH2CH3)2, -CH2NH2, -CH2NHCH3, -CH2N(CH3)2, -CH2NHCH2CH3, -CH2N(CH3)(CH2CH3), -CH2N(CH2CH3)2, -CH2CH2NH2, -CH2CH2NHCH3, -CH2CH2N(CH3 )2, -CH(CH3)N(CH3)2, CH2CH2NHCH2CH3, -CH2CH2N(CH2CH3)2, -CH2CH2CH2NH2, -CH2CH2CH2NHCH3, -CH2CH2CH2N(C H3)2, -CH2CH(NH2)CH3, -CH2CH(CH2NH2)CH3, -CH2CH2CH2CH2NH2, -CH2CH2CH2CH2NHCH3, and -CH2CH2CH2CH2N(CH3)2.

[0138] In a preferred embodiment, R 4 C 1-6 Alkyl groups, optionally selected by one, two, or three independently chosen from halogens, C 1-6 Alkyl, C 1-6 Alkyl group substitution; more preferably, R 4 Selected from -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2OCH3, -CH2CH2OCH3, -CH2CH2OCH2CH3 or -CF3.

[0139] In one embodiment of a compound of formula (I), -XR 4 C 1-6 Alkyl, wherein C 1-6 Alkyl groups having the above pairs as R 4 C 1-6 The alkyl group gives the meaning of the preferred or more preferred embodiments. In an embodiment of a compound of formula (I), -XR 4 -OC 1-6 Alkyl, wherein C 1-6 Alkyl groups having the above pairs as R 4 C 1-6 The alkyl group gives the meaning of the preferred or more preferred embodiments. In an embodiment of a compound of formula (I), -XR 4 -NH-C 1-6 Alkyl, wherein C 1-6 Alkyl groups having the above pairs as R 4 C 1-6 The meaning of the embodiments, preferred or more preferred embodiments given by alkyl groups.

[0140] In one embodiment of a compound of formula (I), R 4 -C 0-3 Alkyl-phenyl, preferably phenyl, wherein the phenyl group is optionally replaced by one, two or three C groups independently selected from halogens, optionally substituted with halogens. 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R cSubstitution of groups 2. In specific embodiments, exemplary substituents include, but are not limited to: F, Cl, Br, I, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2CH(CH3)CH3, -CH2CH2CH2CH3, -C(CH3)(CH3)(CH3), -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CH2CF3, -CH(CF3)CH3, -CH2CH(CF3)CH3, -CH2CH2CH2CF3, -C(CF3)(CH3)(CH 3), -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CH2CH(OH)CH3, -CH2CH(CH2OH)CH3, -CH2CH2CH2CH2OH, -C(CH2OH)(CH3)(CH3), -OH, -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH(CH3)(CH3), -CH2OCH3, -CH2OCH2CH3, -CH2CH2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2OCH3 , -CH2CH(OCH3)CH3, -CH2CH(CH2OCH3)CH3, -CH2CH2CH2CH2OCH3, -NH2, -NHCH3, -NHCH2CH3, -N(CH3)2, -N(CH3)(CH2CH3), -N( CH2CH3)2, -CH2NH2, -CH2NHCH3, -CH2N(CH3)2, -CH2NHCH2CH3, -CH2N(CH3)(CH2CH3), -CH2N(CH2CH3)2, -CH2CH2NH2, -CH2CH2 NHCH3, -CH2CH2N(CH3)2, -CH(CH3)N(CH3)2, CH2CH2NHCH2CH3, -CH2CH2N(CH2CH3)2, -CH2CH2CH2NH2, -CH2CH2CH2NHCH3, -CH2 CH2CH2N(CH3)2, -CH2CH(NH2)CH3, -CH2CH(CH2NH2)CH3, -CH2CH2CH2CH2NH2, -CH2CH2CH2CH2NHCH3, and -CH2CH2CH2CH2N(CH3)2.

[0141] In a preferred embodiment, R 4 It is a phenyl group, with 1, 2, or 3 C atoms independently selected from halogens, optionally substituted by halogens. 1-6 Alkyl and -(CR) c R c ) 0-6 -N(R cSubstitution of groups in 2, exemplary substituents include but are not limited to F, Cl, Br, I, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2CH(CH3)CH3, -CH2CH2CH2CH3, -C(CH3)(CH3)(CH3), -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CH2CF3, -CH(CF3)CH3, -CH2CH(CF3)CH3, -CH2CH2CH2CF3, -C(CF3)(CH3)(CH3), -NH2, -NHCH3, -NHCH2CH3, -N(CH3)2, -N(CH3)(CH2CH3), -N(CH2CH3)2, -CH2NH2, -CH2NHCH3, -CH 2N(CH3)2, -CH2NHCH2CH3, -CH2N(CH3)(CH2CH3), -CH2N(CH2CH3)2, -CH2CH2NH2, -CH2CH2NHCH3, -CH2CH2N(CH3)2, -CH(CH3)N(CH3)2, CH2CH2NHCH2CH3, -CH2CH2N(CH2CH3)2, -CH2CH2CH2NH2, -CH2CH2CH2NHCH3, -CH2CH2CH2N(CH3)2, -CH2CH(NH2)CH3, -CH2CH(CH2NH2)CH3, -CH2CH2CH2CH2NH2, -CH2CH2CH2CH2NHCH3 and -CH2CH2CH2CH2N(CH3)2; more preferably, R 4 Selected from

[0142] In one embodiment of a compound of formula (I), R 4 -C 0-3 Alkyl-phenyl, preferably phenyl, with -(CR c R c ) 0-6 -N(R c )2 replaces, where R c Each independently is H or C 1-6 Alkyl group, wherein the R on N is alkyl c One of them, together with the atoms on the benzene ring it is attached to and adjacent atoms, forms a 4-7 membered nitrogen-containing heterocycle. In a specific embodiment, R 4 Selected from Among them, each R cIt is independently selected from H, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2CH(CH3)CH3, -CH2CH2CH2CH3 and -C(CH3)(CH3)(CH3).

[0143] In a preferred embodiment, R 4 for

[0144] In one embodiment of a compound of formula (I), -XR 4 -C 0-6 Alkyl-phenyl, preferably -C 0-3 alkyl-phenyl, more preferably phenyl, wherein the phenyl has the above pairs as R 4 The phenyl group gives the meaning of the embodiments, preferred or more preferred embodiments. In an embodiment of a compound of formula (I), -XR 4 -OC 0-6 Alkyl-phenyl, preferably -OC 0-3 alkyl-phenyl, more preferably -O-phenyl, wherein the phenyl has the above pairs as R 4 The phenyl group gives the meaning of the embodiments, preferred or more preferred embodiments. In an embodiment of a compound of formula (I), -XR 4 -NH-C 0-6 Alkyl-phenyl, preferably C 0-3 alkyl-phenyl, more preferably -NH-phenyl, wherein the phenyl has the above pairs as R 4 The meaning of the phenyl group given in the embodiments, preferred or more preferred embodiments.

[0145] In one embodiment of a compound of formula (I), R 4 -C 0-3 Alkyl-C 3-6 cycloalkyl, preferably -C 3-6 cycloalkyl group, wherein the cycloalkyl group is optionally replaced by one, two or three C groups independently selected from halogens, optionally substituted with halogens. 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R cSubstitution of groups 2. In specific embodiments, exemplary substituents include, but are not limited to: F, Cl, Br, I, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2CH(CH3)CH3, -CH2CH2CH2CH3, -C(CH3)(CH3)(CH3), -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CH2CF3, -CH(CF3)CH3, -CH2CH(CF3)CH3, -CH2CH2CH2CF3, -C(CF3)(CH3)(CH 3), -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CH2CH(OH)CH3, -CH2CH(CH2OH)CH3, -CH2CH2CH2CH2OH, -C(CH2OH)(CH3)(CH3), -OH, -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH(CH3)(CH3), -CH2OCH3, -CH2OCH2CH3, -CH2CH2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2OCH3 , -CH2CH(OCH3)CH3, -CH2CH(CH2OCH3)CH3, -CH2CH2CH2CH2OCH3, -NH2, -NHCH3, -NHCH2CH3, -N(CH3)2, -N(CH3)(CH2CH3), -N( CH2CH3)2, -CH2NH2, -CH2NHCH3, -CH2N(CH3)2, -CH2NHCH2CH3, -CH2N(CH3)(CH2CH3), -CH2N(CH2CH3)2, -CH2CH2NH2, -CH2CH2 NHCH3, -CH2CH2N(CH3)2, -CH(CH3)N(CH3)2, CH2CH2NHCH2CH3, -CH2CH2N(CH2CH3)2, -CH2CH2CH2NH2, -CH2CH2CH2NHCH3, -CH2 CH2CH2N(CH3)2, -CH2CH(NH2)CH3, -CH2CH(CH2NH2)CH3, -CH2CH2CH2CH2NH2, -CH2CH2CH2CH2NHCH3, and -CH2CH2CH2CH2N(CH3)2.

[0146] In a preferred embodiment, R 4 It is selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; more preferably cyclopropyl.

[0147] In one embodiment of a compound of formula (I), -XR 4 -C 0-6 Alkyl-C3-6 cycloalkyl, preferably -C 0-3 Alkyl-C 3-6 cycloalkyl, more preferably C 3-6 cycloalkyl group, wherein the cycloalkyl group has the above pairs as R 4 The cycloalkyl group in the text indicates the meaning of the embodiments, preferred or more preferred embodiments. In an embodiment of a compound of formula (I), -XR 4 -OC 0-6 Alkyl-C 3-6 cycloalkyl, preferably -OC 0-3 Alkyl-C 3-6 cycloalkyl, more preferably -OC 3-6 cycloalkyl group, wherein the cycloalkyl group has the above pairs as R 4 The cycloalkyl group in the text indicates the meaning of the embodiments, preferred or more preferred embodiments. In an embodiment of a compound of formula (I), -XR 4 -NH-C 0-6 Alkyl-C 3-6 cycloalkyl, preferably -NH-C 0-3 Alkyl-C 3-6 cycloalkyl, more preferably -NH-C 3-6 cycloalkyl group, wherein the cycloalkyl group has the above pairs as R 4 The cycloalkyl groups in the text give the meaning of the embodiments, preferred or more preferred embodiments.

[0148] In one embodiment of a compound of formula (I), R 4 -C 0-3 Alkyl group – a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, preferably a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heterocyclic alkyl group is optionally substituted with 1, 2, or 3 C atoms independently selected from halogens, optionally substituted with halogens. 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R cSubstitution of groups 2. In specific embodiments, exemplary substituents include, but are not limited to: F, Cl, Br, I, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2CH(CH3)CH3, -CH2CH2CH2CH3, -C(CH3)(CH3)(CH3), -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CH2CF3, -CH(CF3)CH3, -CH2CH(CF3)CH3, -CH2CH2CH2CF3, -C(CF3)(CH3)(CH 3), -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CH2CH(OH)CH3, -CH2CH(CH2OH)CH3, -CH2CH2CH2CH2OH, -C(CH2OH)(CH3)(CH3), -OH, -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH(CH3)(CH3), -CH2OCH3, -CH2OCH2CH3, -CH2CH2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2OCH3 , -CH2CH(OCH3)CH3, -CH2CH(CH2OCH3)CH3, -CH2CH2CH2CH2OCH3, -NH2, -NHCH3, -NHCH2CH3, -N(CH3)2, -N(CH3)(CH2CH3), -N( CH2CH3)2, -CH2NH2, -CH2NHCH3, -CH2N(CH3)2, -CH2NHCH2CH3, -CH2N(CH3)(CH2CH3), -CH2N(CH2CH3)2, -CH2CH2NH2, -CH2CH2 NHCH3, -CH2CH2N(CH3)2, -CH(CH3)N(CH3)2, CH2CH2NHCH2CH3, -CH2CH2N(CH2CH3)2, -CH2CH2CH2NH2, -CH2CH2CH2NHCH3, -CH2 CH2CH2N(CH3)2, -CH2CH(NH2)CH3, -CH2CH(CH2NH2)CH3, -CH2CH2CH2CH2NH2, -CH2CH2CH2CH2NHCH3, and -CH2CH2CH2CH2N(CH3)2.

[0149] In one embodiment of a compound of formula (I), R 4The 4-6 membered heterocyclic alkyl group is selected from azirrobutane, oxacyclobutane, thioheterobutyl, pyrrolidinyl (e.g., 1-pyrrolidinyl, 2-pyrrolidinyl and 3-pyrrolidinyl), tetrahydrofuranyl (e.g., 1-tetrahydrofuranyl, 2-tetrahydrofuranyl and 3-tetrahydrofuranyl), tetrahydrothiophenyl (e.g., 1-tetrahydrothiophenyl, 2-tetrahydrothiophenyl and 3-tetrahydrothiophenyl), piperidinyl (e.g., 1-piperidinyl, 2-piperidinyl, 3-piperidinyl and 4-piperidinyl), tetrahydropyranyl (e.g., 4-tetrahydropyranyl), tetrahydrothiophenyl (e.g., 4-tetrahydrothiophenyl), morpholinyl (e.g., morpholino), thiomorpholinyl, dioxane or piperazine.

[0150] In a preferred embodiment, R 4 Selected from

[0151] Optimal R 4 Selected from

[0152] In one embodiment of a compound of formula (I), -XR 4 -C 0-6 Alkyl group - a 4-6 membered heterocyclic alkyl group comprising 1, 2 or 3 heteroatoms independently selected from N, O or S, preferably -C 0-3 Alkyl group – a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, more preferably a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heterocyclic alkyl group has the above pairs as R 4 The meaning of the embodiment, preferred or more preferred embodiment given by the heterocyclic alkyl group. In an embodiment of a compound of formula (I), -XR 4 -OC 0-6 Alkyl group - a 4-6 membered heterocyclic alkyl group containing 1, 2 or 3 heteroatoms independently selected from N, O or S, preferably - OC 0-3 Alkyl group – a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S; more preferably – an O-type 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heterocyclic alkyl group has the above pairs as R 4 The heterocyclic alkyl group in the text indicates the meaning of the embodiments, preferred or more preferred embodiments. In an embodiment of a compound of formula (I), -XR 4 -NH-C 0-6 Alkyl group - a 4-6 membered heterocyclic alkyl group comprising 1, 2 or 3 heteroatoms independently selected from N, O or S, preferably -NH-C 0-3Alkyl group – a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S; more preferably – NH group – a 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heterocyclic alkyl group has the above pairs as R 4 The meaning of the embodiments, preferred or more preferred embodiments given by the heterocyclic alkyl groups.

[0153] In one embodiment of a compound of formula (I), R 4 -C 0-3 Alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, preferably comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heteroaryl group is optionally substituted with 1, 2, or 3 C atoms independently selected from halogens, optionally substituted with halogens. 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R cSubstitution of groups 2. In specific embodiments, exemplary substituents include, but are not limited to: F, Cl, Br, I, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)CH3, -CH2CH(CH3)CH3, -CH2CH2CH2CH3, -C(CH3)(CH3)(CH3), -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CH2CF3, -CH(CF3)CH3, -CH2CH(CF3)CH3, -CH2CH2CH2CF3, -C(CF3)(CH3)(CH 3), -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CH2CH(OH)CH3, -CH2CH(CH2OH)CH3, -CH2CH2CH2CH2OH, -C(CH2OH)(CH3)(CH3), -OH, -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH(CH3)(CH3), -CH2OCH3, -CH2OCH2CH3, -CH2CH2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2OCH3 , -CH2CH(OCH3)CH3, -CH2CH(CH2OCH3)CH3, -CH2CH2CH2CH2OCH3, -NH2, -NHCH3, -NHCH2CH3, -N(CH3)2, -N(CH3)(CH2CH3), -N( CH2CH3)2, -CH2NH2, -CH2NHCH3, -CH2N(CH3)2, -CH2NHCH2CH3, -CH2N(CH3)(CH2CH3), -CH2N(CH2CH3)2, -CH2CH2NH2, -CH2CH2 NHCH3, -CH2CH2N(CH3)2, -CH(CH3)N(CH3)2, CH2CH2NHCH2CH3, -CH2CH2N(CH2CH3)2, -CH2CH2CH2NH2, -CH2CH2CH2NHCH3, -CH2 CH2CH2N(CH3)2, -CH2CH(NH2)CH3, -CH2CH(CH2NH2)CH3, -CH2CH2CH2CH2NH2, -CH2CH2CH2CH2NHCH3, and -CH2CH2CH2CH2N(CH3)2.

[0154] In one embodiment of a compound of formula (I), R 4 The 5-6 heteroaryl groups are selected from pyrrole, furanyl, thiophene, imidazolyl, furazonyl, oxazolyl, oxadiazolyl, oxtriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl, tetrazolyl, pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl, and triazinyl.

[0155] In a preferred embodiment, R 4 Selected from Most preferably, R 4 Selected from

[0156] In one embodiment of a compound of formula (I), -XR 4 -C 0-6 Alkyl group - comprising 1, 2 or 3 heteroatoms independently selected from N, O or S, 5-6 membered heteroaryl group, preferably -C 0-3 Alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, more preferably 5-6-membered heteroaryl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heteroaryl group has the above pairs as R 4 The meaning of the embodiments, preferred or more preferred embodiments given by the heteroaryl group in formula (I). In an embodiment of a compound of formula (I), -XR 4 -OC 0-6 Alkyl group – comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, 5-6 membered heteroaryl group, preferably OC. 0-3 Alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, more preferably 5-6-membered heteroaryl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heteroaryl group has the above pairs as R 4 The meaning of the embodiments, preferred or more preferred embodiments given by the heteroaryl group in formula (I). In an embodiment of a compound of formula (I), -XR 4 -NH-C 0-6 Alkyl group - comprising 1, 2 or 3 heteroatoms independently selected from N, O or S, 5-6 membered heteroaryl group, preferably -NH-C 0-3 Alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, more preferably 5-6-membered heteroaryl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heteroaryl group has the above pairs as R 4 The meaning of the embodiments, preferred or more preferred embodiments given by the heteroaryl group.

[0157] In one embodiment of a compound of formula (I), as R 4 -(CR) of substituents c R c ) 0-6 -SR c 、-(CR c R c ) 0-6 -(CO) 0-1 -ORc 、-(CR c R c ) 0-6 -(CO) 0-1 -N(R c )2 are preferably -(CR) c R c ) 0-6 -SR c 、-(CR c R c ) 0-6 -OR c 、-(CR c R c ) 0-6 -N(R c )2; More preferably -(CR c R c ) 0-3 -SR c 、-(CR c R c ) 0-3 -OR c 、-(CR c R c ) 0-3 -N(R c )2.

[0158] In one embodiment of a compound of formula (I), E is a halogen, preferably F.

[0159] In one embodiment of a compound of formula (I), E is -OR d R d C can be arbitrarily replaced by halogens 1-6 Alkyl, preferably C 1-6 Alkyl groups, such as, but not limited to, methyl, ethyl, propyl, isopropyl, trifluoromethyl, difluoromethyl, trifluoroethyl, etc.

[0160] In one embodiment of a compound of formula (I), R 5 for Among them, at most two of Z, G, Y, B, and D are not C. An example of R... 5 Including but not limited to

[0161] In a preferred embodiment, R 5 for Among them, at most two of Z, Y, B, and D are not C. An example of R... 5 Including but not limited to In a more preferred embodiment, R 5 for Optimal R 5 for

[0162] In a preferred embodiment, R 5 for More

[0163] In one embodiment of a compound of formula (I), R 6 R 7 and R 8 At least one of them is a halogen, preferably F. In a more preferred embodiment, R 6 Let F be the integer part of the integer part, and R be the integer part of the integer part. 7 and R 8 For H.

[0164] In one embodiment of a compound of formula (I), R 6 R 7 and R 8 At least one of them is a halogen-substituted C 1-6 Alkyl group, preferably -CF3.

[0165] In one embodiment of a compound of formula (I), R c For H or C 1-6 alkyl.

[0166] The present invention also provides compounds of formula (II), isomers thereof, pharmaceutically acceptable salts or solvates.

[0167]

[0168] Where R 1 R 2 R 3 R 4 R 6 R 7 R 8 A, E, R b Each has the meaning given above for the compound of formula (I).

[0169] In the preferred embodiment of compound (II),

[0170] A is selected from CR a Or N, where R a Selected from halogens;

[0171] R 1 R 2 and R 3 Each is independently selected from H, halogen, or C. 1-6 alkyl;

[0172] Rb Each time it appears, it is independently selected from H, halogen, CN, or C that is optionally substituted by halogen or CN. 1-6 alkyl;

[0173] X is selected from the bond, -O-, or -NH-;

[0174] R 4 Selected from H, halogens, C 1-6 Alkyl, -C 0-3 Alkyl-C 6-10 Aryl, -C 0-3 Alkyl-C 3-8 cycloalkyl, -C 0-3 Alkyl-C 3-8 Cycloalkenyl, -C 0-3 Alkyl group - a 3- to 8-membered heterocyclic alkyl group containing 1, 2, or 3 independent heteroatoms selected from N, O, or S, -C 0-3 Alkyl group – a 3- to 8-membered heterocyclic alkenyl group containing 1, 2, or 3 independent heteroatoms selected from N, O, or S; -C 0-3 Alkyl group – comprising 1, 2, or 3 independent heteroatoms selected from N, O, or S, of 5-10 membered heteroaryl groups, wherein the alkyl, aryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, and heteroaryl groups are optionally substituted by 1, 2, or 3 groups independently selected from: halogen, C group optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -OR c 、-(CR c R c ) 0-6 -N(R c )2, wherein -(CR) is attached to an aryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, or heteroaryl group. c R c ) 0-6 -N(R c )2. Selectively form a 4-7 member nitrogen-containing heterocycle by means of the group attached to the N, together with the atoms on the cyclic group attached thereto and the adjacent atoms;

[0175] R c Each time it appears, it is independently selected from H or C that is optionally substituted with halogen. 1-6 alkyl;

[0176] E is selected from halogens, -OR d or -N(R) d )2-, where R d Each is independently H or C that is optionally substituted with a halogen. 1-6 alkyl;

[0177] R6 R 7 and R 8 Each is independently selected from H, halogens, and C that is optionally substituted with a halogen. 1-6 alkyl;

[0178] Or its isomers, pharmaceutically acceptable salts or solvates.

[0179] In a preferred embodiment of the compound of formula (II), A is CR a , where R a Selected from F or Cl, with Cl being preferred.

[0180] In a preferred embodiment of the compound of formula (II), R b For H.

[0181] In a preferred embodiment of the compound of formula (II), R b It is not H, and there exists 1 R. b Or there exist 2 R b .

[0182] In a preferred embodiment of the compound of formula (II), -XR 4 For H.

[0183] In a preferred embodiment of the compound of formula (II), -XR 4 Not H.

[0184] In a preferred embodiment of the compound of formula (II), R 6 R 7 and R 8 At least one of them is a halogen, preferably F; more preferably R. 6 Let F be the integer part of the integer part, and R be the integer part of the integer part. 7 and R 8 For H.

[0185] In a preferred embodiment of the compound of formula (II), E is a halogen, preferably F.

[0186] In a preferred embodiment of the compound of formula (II), E is -OR d R d C can be arbitrarily replaced by halogens 1-6 Alkyl, preferably C 1-6 Alkyl groups, such as, but not limited to, methyl, ethyl, propyl, isopropyltrifluoromethyl, difluoromethyl, trifluoroethyl, etc.

[0187] In a preferred embodiment of the compound of formula (II), R 4 Selected from H, halogens, C 1-6 Alkyl, phenyl, C 3-6Cycloalkyl, 4-6-membered heterocycloalkyl comprising 1, 2, or 3 independent heteroatoms selected from N, O, or S, and 5-6-membered heteroaryl comprising 1, 2, or 3 independent heteroatoms selected from N, O, or S, wherein the alkyl, phenyl, cycloalkyl, heterocycloalkyl, and heteroaryl groups are optionally substituted by 1, 2, or 3 groups independently selected from: halogen, C groups optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R c )2, wherein -(CR) is attached to a cycloalkyl, heterocycloalkyl, or heteroaryl group. c R c ) 0-6 -N(R c )2. Selectively form a 4-7 member nitrogen-containing heterocycle by means of the group attached to the N, together with the atoms on the cyclic group attached thereto and the adjacent atoms.

[0188] In a preferred embodiment of the compound of formula (II), R 4 Selected from halogens, preferably F or Cl.

[0189] In a preferred embodiment of the compound of formula (II), R 4 Selected from C 1-6 Alkyl groups, such as methyl, ethyl, propyl, isopropyl, etc., are optionally substituted by one, two, or three groups independently selected from the following: halogen, C-groups optionally substituted with halogens. 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R c )2.

[0190] In a preferred embodiment of the compound of formula (II), R 4 Selected from phenyl, C substituted with halogen or halogen. 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R c )2 replaces.

[0191] In a preferred embodiment of the compound of formula (II), R 4 Selected from C 3-6 Cycloalkyl, optionally substituted by one, two or three groups independently selected from the following: halogen, C group optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R c )2.

[0192] In a preferred embodiment of the compound of formula (II), R 4 Selected from 4-6 membered heterocyclic alkyl groups containing 1, 2, or 3 independent heteroatoms selected from N, O, or S, optionally substituted by 1, 2, or 3 groups independently selected from: halogen, C optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R c )2.

[0193] In a preferred embodiment of the compound of formula (II), R 4 Selected from 5-6-membered heteroaryl groups comprising 1, 2, or 3 independent heteroatoms selected from N, O, or S, optionally substituted by 1, 2, or 3 groups independently selected from: halogen, C optionally substituted with halogen 1-6 Alkyl, -(CR c R c ) 0-6 -OR c and -(CR c R c ) 0-6 -N(R c )2.

[0194] In the above embodiments, the -(CR) group attached to the aryl, cycloalkyl, heterocycloalkyl, or heteroaryl group... c R c ) 0-6 -N(R c )2. Selectively form a 4-7 member nitrogen-containing heterocycle by means of the group attached to the N, together with the atoms on the cyclic group attached thereto and the adjacent atoms.

[0195] In a preferred embodiment of the compound of formula (II), Rc Each time it appears, it is independently selected from H or C. 1-6 alkyl.

[0196] The present invention also provides embodiments of compounds of formula (II), wherein R 1 R 2 R 3 R 4 R 6 R 7 R 8 A, E, R b Each has the meaning of the embodiments, preferred, more preferred or most preferred embodiments of the compound of formula (I) given above.

[0197] It should be noted that the compounds of the present invention cover the various independent embodiments or specific embodiments described above, as well as embodiments consisting of any combination or sub-combination of the various embodiments or specific embodiments described above, and embodiments consisting of any combination of any preferred or exemplary embodiments described above.

[0198] Specific embodiments of the compounds of the present invention include the following specific compounds or isomers thereof, pharmaceutically acceptable salts or solvates.

[0199]

[0200]

[0201]

[0202] The compounds defined above and their various specific embodiments herein are inhibitors of Ras mutations, particularly KRas mutations, including mutations at codons G12, G13, and Q61, such as G12C, G12D, and G13D mutations. The compounds of the present invention, particularly those specifically exemplified in the context herein, have shown inhibitory activity against Ras-mutated, particularly KRas G12C-mutated, cells in cytoassays, as shown in the Activity Examples section below. Therefore, the compounds of the present invention can be used to treat or prevent diseases mediated by Ras mutations, preferably KRas mutations, most preferably KRas G12C mutations, such as diseases or conditions that can be treated by inhibiting Ras mutations, preferably KRas mutations, most preferably KRas G12C mutations, or diseases or conditions in which Ras mutation activity plays a role or is involved, particularly treating or preventing tumors or cancers by inhibiting Ras mutations, preferably KRas mutations, most preferably KRas G12C mutations.

[0203] In addition to exhibiting inhibitory activity against KRas G12C mutations, some compounds of the present invention also exhibit inhibitory activity against KRas G12D mutations, and others exhibit inhibitory activity against KRas G13D mutations.

[0204] In addition to exhibiting Ras mutation and preferably KRas mutation inhibitory activity, the compounds defined herein and their various specific embodiments, especially the compounds in the examples, possess improved structural patterns. Compared with existing KRas mutant protein inhibitors, they retain comparable or enhanced, or even significantly enhanced, KRas mutant protein and related cancer cell proliferation inhibitory activities; they have different bioactivity spectra and can be used for new indications; they have improved metabolic stability, resulting in better pharmacokinetic properties; and they have improved physicochemical properties, thus exhibiting good drug-like properties, such as easier absorption in vivo.

[0205] Based on the above, the present invention also provides technical solutions in the following aspects.

[0206] On the one hand, the present invention provides compounds of the present invention, their isomers, or pharmaceutically acceptable salts or solvates thereof for use as medicines.

[0207] On the other hand, the present invention provides compounds, isomers thereof, or pharmaceutically acceptable salts or solvates thereof for the treatment and / or prevention of diseases mediated by Ras mutations, preferably KRas mutations.

[0208] Pharmaceutical Compositions and Their Administration

[0209] On the other hand, the present invention provides pharmaceutical compositions comprising compounds of formula (I) or formula (II) as defined above, their isomers, or pharmaceutically acceptable salts or solvates thereof, and pharmaceutically acceptable carriers, diluents, or excipients. The pharmaceutical compositions of the present invention can be used to treat or prevent diseases mediated by Ras mutations, particularly KRas mutations, such as KRas G12C, KRas G12D, or KRas G13D mutations, such as tumors or cancers.

[0210] The pharmaceutical compositions of the present invention described above can be formulated using techniques known to those skilled in the art, such as those disclosed in Remington's Pharmaceutical Sciences, 20th edition.

[0211] The administration and delivery of the pharmaceutical compositions of this invention conform to good medical practice. Factors to be considered in this context include the specific disorder being treated, the specific mammal being treated, the individual patient's clinical condition, the cause of the disorder, the location of drug delivery, the method of administration, the administration schedule, and other factors well known to the physician practitioner. The optimal dosage level and frequency of administration of the pharmaceutical compositions of this invention will be determined through clinical trials required in the pharmaceutical field. Typically, for example, the daily dose range for oral administration is from about 0.001 mg to about 100 mg per kg of patient body weight, often from 0.01 mg to about 50 mg per kg of body weight, for example from 0.1 to 10 mg per kg of body weight, preferably from about 0.01 to about 35 mg per kg of body weight, administered as a single dose or in divided doses. For a 70 kg human subject, a suitable dose range is from about 0.07 to about 7000 mg / day, preferably from about 0.7 to about 2500 mg / day. It should be understood that it may be necessary to use doses exceeding these limits in certain circumstances.

[0212] The compositions of the present invention can be administered in any suitable manner, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal, inhalation, epidural, and intranasal administration, and, if local treatment is required, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, oral administration is used.

[0213] The compositions of the present invention can be administered in any convenient form, such as tablets, powders, capsules, lozenges, granules, solutions, dispersants, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. The compositions may contain conventional components of pharmaceutical preparations, such as diluents (e.g., glucose, lactose, or mannitol), carriers, pH adjusters, buffers, sweeteners, fillers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow aids, processing aids, colorants, flavorings, tasters, other known additives, and other active agents. Suitable carriers and excipients are well known to those skilled in the art and detailed in, for example, Ansel, Howard C., et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004.

[0214] Treatment methods and uses

[0215] As described above, the compounds of formula (I) or formula (II) of the present invention and the compounds of various specific embodiments thereof, especially the compounds specifically prepared and characterized in the examples, exhibit inhibitory effects on Ras mutations, especially KRas mutations, such as KRasG12C, KRas G12D or KRas G13D mutations.

[0216] Therefore, on the other hand, the present invention provides a method for inhibiting Ras mutations, especially KRas mutations, preferably KRasG12C, KRas G12D or KRas G13D mutations, and most preferably KRas G12C mutations in cells, comprising contacting cells with a compound of formula (I) or formula (II) of the present invention, its isomers or pharmaceutically acceptable salts or solvates thereof to inhibit the activity of Ras mutations, especially KRas mutations, preferably KRas G12C, KRas G12D or KRas G13D mutations, and most preferably KRas G12C mutations in cells.

[0217] Based on the same properties, the present invention also provides a method for inhibiting abnormal cell growth in mammals, comprising administering to the mammal a therapeutically effective amount of a compound of formula (I) or formula (II) of the present invention, its isomers or pharmaceutically acceptable salts or solvates thereof, or a pharmaceutical composition comprising a compound of formula (I) or formula (II) of the present invention, its isomers or pharmaceutically acceptable salts or solvates thereof.

[0218] On the other hand, the present invention provides a method for treating and / or preventing diseases mediated by Ras mutations, particularly KRas mutations, preferably KRasG12C, KRas G12D or KRas G13D mutations, most preferably KRas G12C mutations, comprising administering to a subject in need a therapeutically effective amount of a compound of formula (I) or formula (II) of the present invention, its isomers or pharmaceutically acceptable salts or solvates thereof, or a pharmaceutical composition comprising a compound of formula (I) or formula (II) of the present invention, its isomers or pharmaceutically acceptable salts or solvates thereof.

[0219] On the other hand, the present invention provides the use of compounds of formula (I) or (II) of the present invention, their isomers or pharmaceutically acceptable salts or solvates thereof, or pharmaceutical compositions comprising compounds of formula (I) or (II) of the present invention, their isomers or pharmaceutically acceptable salts or solvates thereof, for inhibiting Ras mutations in cells, especially KRas mutations, preferably KRas G12C, KRas G12D or KRas G13D mutations, most preferably KRas G12C mutations, or for inhibiting abnormal cell growth in mammals, or for treating and / or preventing diseases mediated by Ras mutations, especially KRas mutations, preferably KRas G12C, KRas G12D or KRas G13D, most preferably KRas G12C mutations.

[0220] On the other hand, the present invention provides the use of compounds of formula (I) or (II) of the present invention, their isomers or pharmaceutically acceptable salts or solvates thereof, or pharmaceutical compositions comprising compounds of formula (I) or (II) of the present invention, their isomers or pharmaceutically acceptable salts or solvates thereof, in the preparation of medicaments for treating and / or preventing diseases mediated by Ras mutations, especially KRas mutations, preferably KRas G12C, KRas G12D or KRas G13D mutations, most preferably KRas G12C mutations.

[0221] With respect to the various methods and applications provided by the present invention, the abnormal cell growth or diseases mediated by Ras mutations, especially KRas mutations, preferably KRas G12C, KRas G12D or KRas G13D, and most preferably KRas G12C mutations, particularly refer to cancer or tumors. Exemplary examples of 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 eye, uterine cancer, ovarian cancer, rectal cancer, anal region cancer, stomach cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small bowel 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 ureter cancer, renal cell carcinoma, renal pelvis cancer, central nervous system tumors (CNS), primary CNS lymphoma, spinal tumors, brainstem glioma, or pituitary adenoma.

[0222] With respect to the various methods and applications provided by the present invention, the abnormal cell growth or diseases mediated by Ras mutations, especially KRas mutations, preferably KRas G12C, KRas G12D or KRas G13D are preferably selected from lung adenocarcinoma, lung cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, and leukemia; most preferably selected from lung adenocarcinoma, colon cancer, rectal cancer, pancreatic cancer, and bile duct cancer.

[0223] Therefore, in a preferred embodiment of this aspect, the present invention provides methods and uses for treating or preventing cancer or tumors by inhibiting KRas-G12C mutations. In a further preferred embodiment, the present invention provides methods and uses for treating or preventing lung adenocarcinoma, colon cancer, rectal cancer, pancreatic cancer, and bile duct cancer by inhibiting KRas-G12C mutations.

[0224] Drug combination

[0225] The compounds of the present invention can be administered as the sole active ingredient or in combination with other drugs or therapies.

[0226] Therefore, on the other hand, the present invention provides pharmaceutical compositions comprising, or consisting of, compounds of formula (I) or formula (II) of the present invention, their isomers or pharmaceutically acceptable salts or solvates thereof, and other active agents, or both. Such pharmaceutical compositions are used to inhibit abnormal cell growth in mammals, or to treat and / or prevent diseases mediated by Ras mutations, preferably KRas mutations.

[0227] The other active agents may be one or more other compounds of the present invention, or may be a second or other (e.g., a third) compound that is compatible with the compounds of the present invention, i.e., does not adversely affect each other, or has complementary activity. For example, these active agents may be compounds known to regulate other biological active pathways, or may be compounds that regulate different components in the biological active pathways involved by the compounds of the present invention, or even compounds that overlap with the biological targets of the compounds of the present invention.

[0228] In one 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, mitosis inhibitors, topoisomerase inhibitors, anti-hormonal drugs, angiogenesis inhibitors, and cytotoxic agents.

[0229] 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 routes of administration. These other active agents can be administered co-administered with the compounds of the present invention in a single pharmaceutical composition, or separately administered in different discrete units, such as combination products, preferably in the form of a pillbox. When administered separately, they can be administered simultaneously or sequentially, with the sequential administration occurring close together or spaced apart in time. They can be prepared and / or formulated by the same or different manufacturers. Furthermore, the compounds of the present invention and other active agents can be (i) before the combination product is sent to a physician (e.g., in the case of a pillbox containing the compounds of the present invention and other drugs); (ii) by the physician themselves (or under the guidance of a physician) before administration; or (iii) by the patient themselves, for example, during the sequential administration of the compounds of the present invention and other active agents, in combination therapy.

[0230] The compounds of the present invention can also be combined with antitumor therapies, including but not limited to surgery, radiation therapy, transplantation (e.g., stem cell transplantation, bone marrow transplantation), tumor immunotherapy, and chemotherapy.

[0231] Therefore, on the other hand, the present invention also provides a pillbox comprising two or more separate pharmaceutical compositions, at least one of which comprises a compound of formula (I) or formula (II) of the present invention, its isomers, or pharmaceutically acceptable salts or solvates thereof, and means for respectively containing said compositions, such as containers, dispensing bottles, or separate foil packages, for example blister packs for packaging tablets, capsules, etc. The pillbox of the present invention is particularly suitable for administering different dosage forms, such as oral and parenteral dosage forms, or for administering different compositions at different dose intervals.

[0232] With respect to the above-described pharmaceutical compositions, drug combinations, or kits of the present invention, the abnormal cell growth involved or the diseases mediated by Ras mutations, especially KRas mutations, preferably KRas G12C, KRas G12D, or KRas G13D, most preferably KRas G12C mutations, are as defined above for the methods and uses of the present invention.

[0233] For the compounds, pharmaceutical compositions, methods, uses, pharmaceutical combinations and kits of the present invention described above, compounds of formula (II) described above, their isomers or pharmaceutically acceptable salts or solvates thereof are preferred, and compounds defined in specific embodiments of formula (II) and the specific compounds listed above, namely compounds 1-25, are more preferred.

[0234] When this document describes a dose of a drug or its pharmaceutically acceptable salt, it should be understood that the dose is based on the weight of the free base and does not include any of its hydrates or solvates, unless the package insert indicates that the dose is based on the weight of the salt, hydrate, or solvate. Attached Figure Description

[0235] Appendix Figure 1 This demonstrates the antitumor activity of the compound from Example 9 in a human non-small cell lung cancer NCI-H358 xenograft mouse model and its effect on body weight.

[0236] Appendix Figure 2 This demonstrates the antitumor activity and effect on body weight of the compound from Example 9 in a BALB / c nude mouse model of human pancreatic cancer Mia PaCa-2 cell subcutaneous xenograft tumor.

[0237] Preparation method of the compound of the present invention

[0238] On the other hand, the present invention also provides a method for preparing the compound defined in the present invention.

[0239] The compounds of formula (I) or (II) of the present invention, their isomers, or their pharmaceutically acceptable salts or solvates can be prepared by a variety of methods, including those given below, those given in the examples, or similar methods. General synthetic schemes for synthesizing the compounds of the present invention are illustrated below.

[0240] For each reaction step of each general synthetic scheme, appropriate reaction conditions are known to those skilled in the art or can be conventionally determined. The method steps for synthesizing the compounds of the present invention can be performed under reaction conditions known per se (including those specifically mentioned), in the absence or generally in the presence of solvents or diluents (including, for example, solvents or diluents that are inert to the reagents used and soluble in them), in the absence or in the presence of catalysts, condensing agents, or neutralizing agents (e.g., ion exchangers, such as cation exchangers, such as H+). + In the case of (form), depending on the nature of the reaction and / or reactants, at reduced, normal or elevated temperatures (e.g., from about -100°C to about 190°C, including, for example, from about -78°C to about 150°C, for example, from about 0°C to about 125°C, room temperature, -20 to 40°C or reflux temperature), at atmospheric pressure or in a closed container, under pressure when appropriate, and / or in an inert atmosphere such as argon or nitrogen.

[0241] Depending on the reactivity of the compounds used, the above reactions are usually carried out at a temperature between room temperature and the boiling temperature of the solvent used.

[0242] The raw materials and reagents used in the preparation of these compounds are generally commercially available, or can be prepared by the methods described below, similar to those described below, or by methods known in the art.

[0243] Unless otherwise specified in the description of the method, solvents suitable for any particular reaction include: those solvents specifically mentioned, or, for example, water; esters, such as lower alkyl esters of lower alkanes, such as ethyl acetate; ethers, such as aliphatic ethers, such as diethyl ether, or cyclic ethers, such as tetrahydrofuran or dioxane; liquid aromatic hydrocarbons, such as benzene or toluene; alcohols, such as methanol, ethanol, or 1- or 2-propanol; nitriles, such as acetonitrile; halogenated hydrocarbons, such as dichloromethane or chloroform; amides, such as dimethylformamide, N-methylpyrrolidone-2-one, or dimethylacetamide; bases, such as heterocyclic nitrogen bases, such as pyridine or triethylamine; carboxylic anhydrides, such as lower alkanes anhydrides, such as acetic anhydride; cyclic, straight-chain, or branched hydrocarbons, such as cyclohexane, hexane, or isopentane; or mixtures of these solvents, such as aqueous solutions. Such solvent mixtures may also be used for post-processing, such as post-processing by chromatography or partitioning.

[0244] If necessary, the raw materials and intermediates in the synthetic reaction process can be separated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, and chromatography. If the intermediates and final products are obtained in solid form, purification can also be performed by recrystallization or aging. The materials can be characterized using conventional methods, including physical constants and spectroscopic data.

[0245] The reaction mixture is post-processed in a conventional manner, such as by mixing with water, separating the phases, and purifying the crude product by chromatography where appropriate.

[0246] Those skilled in the art will recognize the presence of a stereocenter in the compound of formula (I) or formula (II). At all stages of the reaction, the mixture of isomers formed can be separated into individual isomers, such as diastereomers or enantiomers, or into any desired mixture of isomers, such as racemates or mixtures of diastereomers, see, for example, “Stereochemistry of Organic Compounds” by E.L. Leel, S.H. Wilen, and L.N. Mander (Wiley-Interscience, 1994).

[0247] In certain specific cases, it may be necessary to protect specific reactive groups with appropriate protecting groups to avoid side reactions with other reactive groups that may be present in compounds of formula (I) or (II) and may compete with or interfere with the reaction. By way of example only, if one or more groups in a compound of formula (I) or (II) are or contain the groups C(O)OH, NH2, or OH and that group has a similar or even stronger reactivity than the desired reaction position, it is advantageous to protect these groups before the desired reaction occurs. In these cases, it may be necessary to perform additional deprotection steps to remove these protecting groups after the desired reaction has been completed. Suitable protecting groups and methods for protecting and deprotecting different substituents with such suitable protecting groups are well known to those skilled in the art; examples can be found in T. Greene and P. Wuts, Protective Groups in Organic Synthesis (3rd ed.), John Wiley & Sons, NY (1999).

[0248] The present invention also relates to preparation methods in which a compound obtainable as an intermediate in any step of the various preparation methods and processes described below is used as a starting material and the remaining method steps are carried out, or in which the starting material is formed in situ under reaction conditions or used as a derivative, for example, in a protected form or in salt form, or a compound obtainable according to the method of the present invention is generated under the method conditions and further treated in situ.

[0249] Synthesis Scheme I:

[0250] The compounds of the present invention can be prepared according to the following exemplary schemes or similarly, wherein, unless otherwise stated, the variables are as defined above.

[0251]

[0252] This invention prepares compounds of formula (I) or (II) via synthetic scheme I. In step A, compound 2 is obtained through the chlorination reaction of an aromatic compound. Then, compound 3 is obtained through methyl esterification in step B, followed by amino acid condensation in step C to obtain compound 4. Compound 4 is cyclized under basic conditions to obtain compound 5, which is then chlorinated to obtain intermediate IntA. The key intermediate IntA reacts with a piperazine or a piperazine derivative with a single protecting group under basic conditions via nucleophilic substitution to obtain intermediate IntB. The latter is then reacted via aromatic nucleophilic substitution (when X = O) or metal-catalyzed coupling (when X = directly connected) to obtain compound 6. In step H, compound 6 is catalytically coupled to introduce a protecting group R... 5The group yields compound 7. Compound 7 is deprotected to yield compound 8, which is then acylated with an acyl chloride or carboxylic acid to give compound of general formula I.

[0253] The typical reaction conditions and reagents used in the chlorination, esterification, condensation, cyclization, nucleophilic substitution, catalytic coupling, and acylation reactions involved in Synthesis Scheme 1 are well known in the art and fall within the scope of conventional experience of those skilled in the art. Alternatively, they can be determined by those skilled in the art by making appropriate modifications based on the typical conditions of such reactions in the art, and based on the characteristics of the raw materials and the target product.

[0254] Synthesis Scheme II:

[0255] Among them -XR 4 The compounds of the present invention with the form H can be synthesized according to the following exemplary general schemes.

[0256]

[0257] In synthetic scheme II, intermediate Int B is subjected to catalytic reductive dechlorination to obtain intermediate Int C, which is then subjected to the same reaction steps H, I, and J as in scheme I to obtain -XR. 4 The compound of this invention is H.

[0258] Synthesis Scheme III:

[0259] The compounds of the present invention where A is N can be synthesized according to the following exemplary general schemes.

[0260]

[0261] In this synthetic scheme, compound 11 undergoes an iodination reaction to yield compound 12, which in turn undergoes a metal-catalyzed carbonyl insertion reaction to yield compound 13. Starting from compound 13, the compound of the present invention, wherein A is N, is obtained through the same steps C-J as in Scheme I.

[0262] Synthesis Scheme IV:

[0263] Where A is N and -XR 4 The compounds of the present invention with the form H can be synthesized according to the following exemplary general schemes.

[0264]

[0265] In this synthetic scheme, starting from the intermediate Int E, the method is the same as in Scheme II, to synthesize where A is N and -XR. 4 The compound of this invention is H.

[0266] In the above synthetic schemes II, III, and IV, The typical reaction conditions and reagents used in each of the reactions involved are well known in the art and fall within the scope of the conventional experience of those skilled in the art, or can be determined by those skilled in the art based on the typical conditions of such reactions in the art, and based on the characteristics of the raw materials and the target product.

[0267] Synthesis Examples

[0268] The present invention will be further described below with reference to embodiments. It should be noted that the following embodiments are exemplary and should not be regarded as limiting the scope of protection of the present invention.

[0269] In the description of the implementation schemes and subsequent specific embodiments, the following abbreviations are used:

[0270] Boc (tert-butoxycarbonyl); n-BuLi (n-butyllithium); t-BuOK (potassium tert-butoxide); CDCl3 (deuterated chloroform); DCM (dichloromethane); DIEA (N,N-diisopropylethylamine); DME (ethylene glycol dimethyl ether); DMF (N,N-dimethylformamide); DMSO (dimethyl sulfoxide); DMSO-d6 (hexadeuterated dimethyl sulfoxide); EA (ethyl acetate); EDTA-K2 (dipotassium ethylenediaminetetraacetate); EtOH (ethanol); FCC (fast column chromatography); g (gram); h (hour); HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethyl) Urea hexafluorophosphate); HCl (hydrogen chloride); HLM (human liver microsomes); H2O (water); H2SO4 (sulfuric acid); IV (intravenous administration); K2CO3 (potassium carbonate); KOH (potassium hydroxide); LCMS (liquid chromatography-mass spectrometry); LC-MS / MS (liquid chromatography-mass spectrometry-mass spectrometry); MeCN (acetonitrile); MeOH (methanol); Methanol-d4 (tetradeuterated methanol); mg (milligram); MHz (megahertz); min (minute); mL (milliliter); mmol (millimole); m / z (mass-to-charge ratio); N2 (nitrogen); NaCl (sodium chloride); NaH (sodium hydride); NaHCO3 3 (Sodium bicarbonate); NaOH (Sodium hydroxide); Na2SO3 (Sodium sulfite); Na2SO4 (Sodium sulfate); NCCH2CO2H (2-cyanoacetic acid); NCS (chlorosuccinimide); NH4Cl (Ammonium chloride); NIS (iodosuccinimide); NMI (N-methylimidazolium); NMP (N-methylpyrrolidone); NMR (Nuclear magnetic resonance); Pd / C (Palladium on carbon); Pd2(dba)3 (Tris(dibenzylacetone)dipalladium); Pd(dppf)Cl2 (1,1'-bis(diphenylphosphine)ferrocene palladium dichloride); Pd(PPh3)4 (tetraphenylphosphine palladium); PE (petroleum ether); P EG (polyethylene glycol); PO (oral administration); POCl3 (phosphorus oxychloride); rt (room temperature); SiO2 (silica gel); TCFH (N,N,N',N'-tetramethylchloromethanesulfonyl hexafluorophosphate); TEA (triethylamine); TFA (trifluoroacetic acid); THF (tetrahydrofuran); TLC (thin-layer chromatography); TsOH (p-toluenesulfonic acid); TsOH·H2O (p-toluenesulfonic acid monohydrate); μL (microliter); μM (micromolar concentration); μmol (micromolar); v / v (volume ratio); w / w (mass ratio); Xantphos (4,5-bisdiphenylphosphine-9,9-dimethyloxanthracene).

[0271] In the following examples, the names and structures of the synthesized target compounds are given. Any discrepancies between the names and structures are not intentional; in this case, the structure is definitive.

[0272] Experimental methods not specifically described in the following examples are generally performed under standard conditions for such reactions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight. Unless otherwise stated, liquid ratios are volume ratios.

[0273] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial sources, prepared according to existing methods, or prepared according to methods similar to those disclosed in this application.

[0274] In the following embodiments, 1 ¹H-NMR spectra were recorded using a Bruker (400 MHz) instrument, and chemical shifts are expressed as δ (ppm) relative to the deuterated solvent peaks (CDCl₃: δ = 7.26 ppm; CD₃OD: δ = 3.31 ppm; DMSO-d₆: δ = 2.50 ppm). Mass spectra were recorded using an Aglient 1100 liquid chromatograph + Aglient G6100 LCMS system.

[0275] Synthesis of intermediate a

[0276]

[0277] (2-((tert-Butoxycarbonyl)amino)-7-fluorobenzothiazo-4-yl)boronic acid

[0278]

[0279] Step A: N-((2-bromo-5-fluorophenyl)aminomethanesulfonyl)benzamide

[0280] In a round-bottom flask equipped with a magnetic stir bar, benzoyl isothiocyanate (28 g, 210 mmol) was dissolved in THF (300 mL). After cooling in an ice bath, a THF solution of 2-bromo-5-fluoroaniline (40 g, 210 mmol) was added dropwise to the system with stirring. After the addition was complete, the reaction was stirred for 1 h. The reaction was confirmed by LCMS to be complete. The crude product was obtained after concentration under reduced pressure and directly used in the next step. LCMS (m / z): 353.1 (M+H).

[0281] Step B: 1-(2-bromo-5-fluorophenyl)thiourea

[0282] Under stirring at room temperature, a solution of NaOH (16.8 g, 420 mmol) in water (150 mL) was added to a solution of N-((2-bromo-5-fluorophenyl)aminomethanesulfonyl)benzamide (the crude product obtained in step A) in THF (300 mL), and the mixture was heated to 80 °C for 16 h. After concentration under reduced pressure, the solution was diluted with EA (200 mL), washed with water (150 mL), and the aqueous and organic phases were separated. The aqueous phase was then extracted twice with EA (100 mL × 2). The organic phases were combined, washed with saturated sodium chloride aqueous solution, and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to obtain the crude product. 200 mL of petroleum ether was added and the mixture was stirred and filtered to obtain a white solid 1-(2-bromo-5-fluorophenyl)thiourea (40 g, yield 77%). LCMS (m / z): 249.0, 251.0 (M+H).

[0283] Step C: 4-Bromo-7-fluorobenzo[d]thiazol-2-amine hydrobromide

[0284] With stirring, a solution of liquid bromine (23 g, 141 mmol) in chloroform (70 mL) was slowly added dropwise to a solution of 1-(2-bromo-5-fluorophenyl)thiourea (35 g, 141 mmol) in chloroform (300 mL) cooled in an ice bath. After the addition was complete, the ice bath was removed, and the system was heated to 70 °C and reacted for 48 h. After the reaction was complete, the solution was concentrated under reduced pressure, slurried with EA (100 mL), and filtered to give a white solid 4-bromo-7-fluorobenzo[d]thiazol-2-amine hydrobromide (30.5 g, yield 66%). LCMS (m / z): 247.0, 249.0 (M+H).

[0285] Step D: (4-bromo-7-fluorobenzo[d]thiazolyl)tert-butyl carbamate

[0286] At room temperature, a solution of di-tert-butyl dicarbonate (61 g, 280 mmol) in DCM (60 mL) was slowly added to a solution of 4-bromo-7-fluorobenzo[d]thiazol-2-amine hydrobromide (30 g, 93 mmol) and 4-dimethylaminopyridine (14 g, 112 mmol) in DCM (300 mL). The reaction was allowed to proceed for 3 h at room temperature, and the reaction was detected as complete by LCMS. The reaction solution was concentrated under reduced pressure, diluted with EA (200 mL), washed with water (150 mL), and the aqueous and organic phases were separated. The aqueous phase was then extracted twice with EA (200 mL × 2). The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and slurried with petroleum ether and ethyl acetate (20:1). The mixture was filtered to give a white solid (18 g, 56% yield) of tert-butyl 4-bromo-7-fluorobenzo[d]thiazol-2-yl)carbamate. 1H NMR (400MHz, Chloroform-d) δ9.29 (s, 1H), 7.54 (dd, J = 8.6, 4.8Hz, 1H), 6.94–6.81 (m, 1H), 1.52 (s, 9H). LCMS (m / z): 291.0, 293.0 (M+H).

[0287] Step E: (2-((tert-Butoxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boronic acid

[0288] Under nitrogen protection and stirring, NaH (60% w / w, 3 g, 75.6 mmol) was added to a THF (200 mL) solution of (17.5 g, 50.4 mmol) tert-butyl carbamate (4-bromo-7-fluorobenzo[d]thiazolyl-2-yl)carbamate cooled in an ice bath. Stirring was continued at this temperature for 30 min, then transferred to a -68 °C dry ice / ethanol bath. Then, n-butyllithium (30.2 mL, 75.6 mmol, 2.5 M / THF) was slowly added, and the mixture was stirred for 10 min. Trimethyl borate (156 g, 151 mmol) was then added, and the reaction was continued for another 30 min. The reaction was quenched with saturated ammonium chloride aqueous solution (50 mL), diluted with EA (200 mL), washed with water (150 mL), and the aqueous and organic phases were separated. The aqueous phase was then extracted twice with EA (100 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was added to petroleum ether and slurried, then filtered to obtain a white solid (2-((tert-butoxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boronic acid (10.7 g, yield 68%). 1 H NMR (400MHz, DMSO-d6) δ12.25 (s, 1H), 8.50 (s, 2H), 7.84 (dd, J = 8.0, 6.4Hz, 1H), 7.22–7.13 (m, 1H), 1.54 (s, 9H). LCMS (m / z): 313.2 (M+H).

[0289] Synthesis of intermediate Int A

[0290]

[0291] 7-Bromo-2,4,6-trichloro-8-fluoroquinoline-3-carbamate

[0292]

[0293] Step A: 2-Amino-4-bromo-5-chloro-3-fluorobenzoic acid

[0294] NCS (13.7 g, 102.6 mmol) was added in portions to DMF (140 mL) containing 2-amino-4-bromo-3-fluorobenzoic acid (20 g, 85.5 mmol). The reaction mixture was then heated to 75 °C and stirred at this temperature for 20 h. After cooling to room temperature, the mixture was poured into 700 mL of an ice-water mixture, causing a precipitate to form. The solid was collected by filtration and washed with water (500 mL) and petroleum ether (150 mL), respectively. The product was dried under vacuum to give a pale yellow solid of 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (20 g, yield 87%). LCMS (m / z): 268.0 (M+H).

[0295] Step B: Methyl 2-amino-4-bromo-5-chloro-3-fluorobenzoate

[0296] 2-Amino-4-bromo-5-chloro-3-fluorobenzoic acid (20 g, 74.5 mmol) was dispersed in methanol (150 mL). The mixture was cooled in an ice-water bath, and thionyl chloride (50 mL) was added dropwise to the reaction solution with stirring. After the addition was complete, the reaction mixture was placed at 65 °C and reacted for 20 h. After the reaction was complete as monitored by TLC, the solvent was concentrated to dryness to obtain the target product, methyl 2-amino-4-bromo-5-chloro-3-fluorobenzoate (15.8 g, yield 75%). LCMS (m / z): 282.0 (M+H).

[0297] Step C: Methyl 4-bromo-5-chloro-2-(2-cyanoacetamido)-3-fluorobenzoate

[0298] At room temperature, methyl 2-amino-4-bromo-5-chloro-3-fluorobenzoate (15.8 g, 55.9 mmol), TCFH (18.8 g, 67.1 mmol), and cyanoacetic acid (5.2 g, 61.3 mmol) were dissolved in acetonitrile (100 mL). N-methylimidazole (6.7 mL, 83.9 mmol) was added with stirring. The reaction mixture was heated to 60 °C and stirred for 20 h. After the reaction was complete as monitored by TLC, the reaction solution was cooled to room temperature and poured into water (500 mL), resulting in a white precipitate. The solid was collected by filtration and washed with water (500 mL) and ethyl acetate / petroleum ether (1:4, 150 mL), respectively. After vacuum drying, a white solid of methyl 4-bromo-5-chloro-2-(2-cyanoacetamido)-3-fluorobenzoate (15 g, 76% yield) was obtained. LCMS (m / z): 349.0 (M+H).

[0299] Step D: 7-Bromo-6-chloro-8-fluoro-2,4-dihydroxyquinoline-3-carbamate

[0300] Methyl 4-bromo-5-chloro-2-(2-cyanoacetamido)-3-fluorobenzoate (6 g, 17.2 mmol) was dispersed in THF (120 mL). Potassium tert-butoxide (3.7 g, 32.6 mmol) was added in portions with stirring in an ice bath. After the addition was complete, the reaction mixture was stirred in an ice bath for 1 h. The reaction was monitored by LCMS until complete. After removing the solvent by vacuum distillation, water (200 mL) was added to the system, and the pH of the mixture was adjusted to 7 with 1 N hydrochloric acid. The mixture was filtered and the solid was collected. The obtained solid was washed with water (100 mL) and petroleum ether (150 mL), and dried under vacuum to give a white solid, 7-bromo-6-chloro-8-fluoro-2,4-dihydroxyquinoline-3-carbamate (5.4 g, 99% yield). LCMS (m / z): 317.0 (M+H).

[0301] Step E: 7-Bromo-2,4,6-trichloro-8-fluoroquinoline-3-carbamate

[0302] 7-Bromo-6-chloro-8-fluoro-2,4-dihydroxyquinoline-3-carbamate (5.4 g, 17 mmol) was dispersed in phosphorus oxychloride (100 mL) with stirring at room temperature. The system was heated to 120 °C and stirred for 24 h. After cooling to room temperature, most of the phosphorus oxychloride was removed by vacuum concentration, and then acetonitrile (20 mL) was added. The acetonitrile solution of the product was slowly poured into water (250 mL), precipitating a yellow solid. The solid was filtered and collected, washed with water (200 mL) and petroleum ether (100 mL), and dried under vacuum to give a yellow solid 7-bromo-2,4,6-trichloro-8-fluoroquinoline-3-carbamate (5.4 g, 90% yield). LCMS (m / z): 353.0 (M+H).

[0303] Synthesis of intermediate Int B1

[0304]

[0305] (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester

[0306]

[0307] DIEA (7.3 mL, 44 mmol) and (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (4.4 g, 22 mmol) were added sequentially to a THF (100 mL) solution of 7-bromo-2,4,6-trichloro-8-fluoroquinoline-3-carbamate (intermediate Int A, 7.8 g, 22 mmol) under stirring at room temperature. The resulting mixture was stirred for 1 h at room temperature. After the reaction was complete, the sample was concentrated by rotary evaporation. The crude product was washed with a PE / THF mixed solvent (PE / THF = 10:1, 500 mL) to give a yellow solid (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (9.0 g, yield 79%). LCMS (m / z): 519.2 (M+H).

[0308] Synthesis of intermediate Int B2

[0309]

[0310] 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0311] The synthesis of intermediate Int B2 was performed as described in intermediate Int B1, with piperazine-1-carboxylic acid tert-butyl ester used instead of (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester. LCMS (m / z): 503.2 (M+H).

[0312] Synthesis of intermediate Int C1

[0313]

[0314] (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester

[0315]

[0316] In a round-bottom flask equipped with a magnetic flask, (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate Int B1, 206 mg, 0.4 mmol), Pd(PPh3)4 (23 mg, 20 μmol), formic acid (55 mg, 1.2 mmol), and triethylamine (80.5 mg, 0.8 mmol) were added. After purging with nitrogen three times, DMF (4 mL) was added. The reaction system was heated to 50 °C and stirred for 6 h. After the reaction was completed, the sample was concentrated using a rotary evaporator. The crude product was purified by FCC (SiO2, EA / PE = 0-50%) to obtain (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (140 mg, yield 73%). LCMS(m / z):485.3(M+H).

[0317] Synthesis of intermediate Int C2

[0318]

[0319] 4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0320] The synthesis of intermediate Int C2 was carried out as described in intermediate Int C1, with (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate B1) replaced by 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate B1). LCMS (m / z): 459.2, 471.2 (M+H).

[0321] Synthesis of intermediate Int D

[0322]

[0323] 2,4-Dichloro-7-bromo-8-fluoro-1,6-naphthidine-3-carbamate

[0324]

[0325] Step A: 2-Chloro-3-fluoro-5-iodopyridine-4-amine

[0326] In a round-bottom flask at room temperature, 2-chloro-3-fluoropyridin-4-amine (3 g, 20.47 mmol) and NIS (6.91 g, 30.71 mmol) were dissolved in 30 mL of acetonitrile. With stirring, TsOH·H₂O (400 mg, 2 mmol) was added. The reaction mixture was heated to 70 °C and reacted for 16 h. After the reaction was complete as monitored by TLC, the reaction mixture, cooled to room temperature, was poured into 100 mL of saturated brine and extracted with ethyl acetate (25 mL × 3). The combined organic phases were washed with saturated Na₂SO₃ aqueous solution and saturated brine, dried over anhydrous Na₂SO₄, and the solvent was concentrated to give a pale yellow solid, 2-chloro-3-fluoro-5-iodopyridin-4-amine (5.4 g, 97% yield). LCMS (m / z): 273.0 (M+H). Step B: Ethyl 4-amino-6-chloro-5-fluoronicotinic acid.

[0327] Compound 2-chloro-3-fluoro-5-iodopyridin-4-amine (5.4 g, 19.82 mmol), Pd(dppf)Cl2 (695 mg, 0.991 mmol), and triethylamine (555 mg, 49.6 mmol) were dispersed in anhydrous ethanol and reacted at 90 °C for 40 h under a carbon monoxide atmosphere. After the reaction was complete, the reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated and purified by FCC (SiO2, EA / DCM = 0-100%) to give a yellow solid ethyl 4-amino-6-chloro-5-fluoronicotinic acid (3.5 g, yield 81%). LCMS (m / z): 219.0 (M+H).

[0328] Step C: Ethyl 6-chloro-4-(2-cyanoacetamido)-5-fluoronicotinic acid

[0329] Cyanoacetic acid (778 mg, 9.15 mmol) and trifluoroacetic anhydride (1.44 g, 6.86 mmol) were dissolved in acetonitrile (5 mL). After stirring at 55 °C for 2 h, the reaction solution and ethyl 4-amino-6-chloro-5-fluoronicotinic acid (1.0 g, 4.57 mmol) were placed in a 20 mL microwave-safe tube and microwave-treated at 105 °C for 2 h. After the reaction was complete, the reaction solution was poured into 25 mL of water, and ethyl acetate (15 mL) was added. The organic phase was washed with water, saturated sodium bicarbonate solution, and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated in solvent, and purified by FCC (SiO2, THF / DCM = 0-40%) to obtain a pale yellow solid, ethyl 6-chloro-4-(2-cyanoacetamido)-5-fluoronicotinic acid (550 mg, yield 42%). LCMS (m / z): 286.0 (M+H).

[0330] Step D: 7-Chloro-8-fluoro-2,4-dihydroxy-1,6-naphthidine-3-carbamate

[0331]

[0332] Ethyl 6-chloro-4-(2-cyanoacetamido)-5-fluoronicotinic acid (300 mg, 1.05 mmol) was dissolved in THF (10 mL) and stirred for 2 min in an ice bath. Potassium tert-butoxide (236 mg, 2.10 mmol) was then slowly added to the reaction solution. The mixture was stirred for 1 h under conditions of natural temperature recovery in an ice bath. After the reaction was complete as determined by LCMS, the reaction solution was added dropwise to a stirred saturated NH4Cl solution (50 mL). The pH of the mixture was adjusted to 7 with 1 N hydrochloric acid. During this process, a white flocculent precipitate formed. The precipitate was filtered, and the solid was collected. The solid was washed with water (10 mL) and petroleum ether (10 mL), and dried under vacuum to obtain a white solid, 7-chloro-8-fluoro-2,4-dihydroxy-1,6-naphthidine-3-carbamate (200 mg, yield 79%). LCMS (m / z): 240.1 (M+H).

[0333] Step E: 2,4,7-Trichloro-8-fluoro-1,6-naphthidine-3-carbamate

[0334]

[0335] 200 mg (0.83 mmol) of 7-chloro-8-fluoro-2,4-dihydroxy-1,6-naphthidine-3-carbamate was dissolved in POCl3 (15 mL). The reaction mixture was heated to 120 °C and stirred overnight. The reaction was monitored by TLC until completion. After cooling to room temperature, the mixture was concentrated under vacuum to remove most of the phosphorus oxychloride, and then acetonitrile (1 mL) was added. The acetonitrile solution of the product was poured into a saturated NaHCO3 aqueous solution (30 mL), and a yellow solid was produced. The solid was filtered, collected, and washed with water (10 mL) and petroleum ether (10 mL), respectively. After vacuum drying, a yellow solid 2,4,7-trichloro-8-fluoro-1,6-naphthidine-3-carbamate (190 mg, yield 82%) was obtained.

[0336] Synthesis of intermediate Int E1

[0337]

[0338] 4-(2,7-dichloro-3-cyano-8-fluoro-1,6-naphthid-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0339] The synthesis of intermediate Int E1 was carried out as described in intermediate Int B1, with 2,4,7-trichloro-8-fluoro-1,6-naphthidium-3-carbamate (intermediate Int D) replacing 7-bromo-2,4,6-trichloro-8-fluoroquinoline-3-carbamate (intermediate Int A), and piperazine-1-carboxylic acid tert-butyl ester replacing (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester. LCMS (m / z): 426.3 (M+H).

[0340] Synthesis of intermediate Int F

[0341]

[0342] (S)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinolin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester

[0343] The synthesis of intermediate Int F was carried out as described in intermediate Int B1, with (S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester salt replaced by (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester. LCMS (m / z): 576.3, 578.3 (M+H).

[0344] Example 1

[0345]

[0346] 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-3-carbamate

[0347] Step A: (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester

[0348] NaH (60% w / w, 9.3 mg, 0.23 mmol) was added to a 2 mL solution of (3R,4R)-4-methoxy-1-methylpyrrolidone-3-ol (27.8 mg, 0.21 mmol) in THF at room temperature with stirring. The mixture was stirred for another 10 min at room temperature. Then, (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinolin-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate B1, 110 mg, 0.21 mmol) was added in a single batch, and the mixture was stirred for 1 h at room temperature. LCMS monitoring showed that intermediate B1 remained. Then, a 1 mL solution of NaH (60% w / w, 5 mg, 0.12 mmol) and (3R,4R)-4-methoxy-1-methylpyrrolidone-3-ol (10 mg, 0.78 mmol) in THF was added again after stirring for 5 min. The resulting mixture was stirred for another 30 min. The reaction was monitored by LCMS until completion. The reaction was quenched with saturated NH4Cl aqueous solution (2 mL), diluted with EA (30 mL), and the resulting organic phase was washed with saturated NaCl (20 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated to obtain crude (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (130 mg, crude product), which was directly added to the next step. LCMS (m / z): 612.4 (M+H).

[0349] Step B: (2R)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester

[0350] At room temperature, a solution of (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidone-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (170 mg, 0.28 mmol), (2-((tert-butoxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boric acid (112 mg, 0.36 mmol), Pd(dppf)Cl2 (24 mg, 0.03 mmol), and K3PO4 (104 mg, 0.49 mmol) in 1,4-dioxane / H2O (v / v = 3:1, 1.6 mL) was added to a microwave-safe reaction tube containing a magnetic induction magnet. After purging the system with nitrogen for 10 min, the reaction tube was sealed and microwave-heated to 100 °C for 1 h. Cool to room temperature, dilute with EA (20 mL), wash with saturated NaCl aqueous solution (20 mL × 3), dry with anhydrous Na2SO4, concentrate under reduced pressure, and preliminarily purify with FCC (SiO2, MeOH / DCM = 0-20%) to obtain (2R)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (150 mg, 62% purity), which was directly used in the next step of the reaction. LCMS (m / z): 800.6 (M+H).

[0351] Step C: 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)-4-((R)-3-methylpiperazin-1-yl)quinoline-3-carbamate

[0352] Under stirring at room temperature, 1 mL of TFA was added dropwise to 2 mL of DCM solution containing (2R)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (150 mg, 62% purity). The resulting mixture was stirred at room temperature for 30 min, and the reaction was monitored by LCMS to indicate completion. The solvent TFA was removed by vacuum concentration, and a small amount of DCM was added for further vacuum concentration. This process was repeated three times to remove residual TFA. The resulting crude product was directly used in the next step without further purification.

[0353] Step D: 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-3-carbamate

[0354] The crude 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidone-3-yl)oxy)-4-((R)-3-methylpiperazin-1-yl)quinoline-3-carbamate obtained in step C was dissolved in DCM (2 mL), cooled in an ice-water bath, and DIEA (103 μL, 0.62 mmol) was added. After stirring for 5 min, a solution of acryloyl chloride (10 μL, 0.12 mmol) in DCM (0.1 mL) was slowly added dropwise. After the addition was complete, the mixture was stirred for 20 min in an ice-water bath. The reaction was monitored by LCMS until it was complete. H2O (5 mL) and EA (30 mL) were added to the system. The aqueous phase was removed by separation. The organic phase was washed with saturated NaCl aqueous solution (20 mL × 3), dried over anhydrous Na2SO4, concentrated, and purified by preparative high performance liquid chromatography to obtain a white solid 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-3-carbamate (15.2 mg, total yield of four steps 7.5%). 1 H NMR(400MHz, Methanol-d4)δ8.12–7.99(m,1H),7.33–7.22(m,1H),7.07–6.97(m,1H),6.87 (dd,J=16.7,10.7Hz,1H),6.30(d,J=16.8Hz,1H),5.83(d,J=10.7Hz,1H),5.65(s,1H),4.24 –4.20(m,1H),4.09–3.89(m,2H),3.83–3.77(m,1H),3.62–3.48(m,5H),3.43–3.38(m,2H),3 .26–3.18(m,2H),3.09–3.03(m,1H),2.98–2.92(m,1H),2.60(s,3H),1.46(d,J=6.8Hz,3H). 19 F NMR (376MHz, Methanol-d4) δ-114.94,-120.48. LCMS (m / z): 654.5 (M+H).

[0355] Example 2

[0356]

[0357] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-3-carbamate

[0358] The synthesis of Example 2 was carried out in accordance with the method described in Example 1, except that in step A, 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate B2) was used instead of (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate B1). 1 H NMR (400MHz, DMSO-d6) δ8.02–7.87(m,3H),7.24(dt,J=8.4,5.1Hz,1H),7.07(t,J=8.8Hz,1H),6. 90(dd,J=16.7,10.5Hz,1H),6.19(dd,J=16.7,2.4Hz,1H),5.75(dd,J=10.4,2.4Hz,1H),5.39–5.3 1(m,1H),4.06–3.96(m,1H),3.94–3.81(m,4H),3.75–3.61(m,4H),3.32(s,3H),3.08–3.00(m,1H) ,2.92(dd,J=11.0,6.2Hz,1H),2.73–2.64(m,1H),2.33(dt,J=9.3,4.3Hz,1H),2.26–2.21(m,3H). 19 F NMR (376MHz, DMSO-d6) δ-112.27 (d, J=4.4Hz), -119.59 (d, J=22.4Hz). LCMS (m / z): 640.5 (M+H).

[0359] Example 3

[0360]

[0361] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-(3-((dimethylamino)methyl)phenyl)-8-fluoroquinoline-3-carbamate

[0362] Step A: 4-(7-bromo-6-chloro-3-cyano-2-(3-((dimethylamino)methyl)phenyl)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0363] Compound 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate B2, 300 mg, 0.60 mmol), (3-((dimethylamino)methyl)phenyl)boronic acid (108 mg, 0.60 mmol), Pd(PPh3)4 (35 mg, 0.03 mmol), K2CO3 (166 mg, 1.2 mmol), acetonitrile (3 mL), and water (1 mL) were added to a microwave tube. After purging with nitrogen three times, the tube was microwave-heated to 40 °C and reacted for 1 h. After cooling to room temperature, the reaction solution was concentrated, and the crude product was purified by FCC (SiO2, EA / PE = 0-60%) to obtain a yellow solid 4-(7-bromo-6-chloro-3-cyano-2-(3-((dimethylamino)methyl)phenyl)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (260 mg, yield 72%). LCMS (m / z): 602.4 (M+H).

[0364] Steps B to D: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-(3-((dimethylamino)methyl)phenyl)-8-fluoroquinoline-3-carbamate

[0365] The subsequent synthesis of Example 3 was carried out in accordance with the synthesis described in Example 1, except that in step B, 4-(7-bromo-6-chloro-3-cyano-2-(3-(((dimethylamino)methyl)phenyl)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester was used instead of (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester. 1 HNMR(400MHz,DMSO-d6)δ8.11-8.10(m,1H),7.92(s,2H),7.81-7.70(m,2H) ,7.53-7.46(m,2H),7.31-7.27(m,1H),7.11-7.09(m,1H),6.91(dd,J=16.6, 10.4Hz,1H),6.22-6.17(m,1H),5.77-5.74(m,1H),3.98-9.84(m,4H),3.83- 3.70(m,4H),3.51-3.46(m,2H),2.24-2.13(m,6H).LCMS(m / z):645.5(M+H).

[0366] Example 4

[0367]

[0368] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-(1-(2-(dimethylamino)ethyl)-1H-pyrazol-5-yl)-8-fluoroquinoline-3-carbamate

[0369] The synthesis of Example 4 was carried out in accordance with the method described in Example 3, except that (1-(2-(dimethylamino)ethyl)-1H-pyrazol-5-yl)boronic acid was used instead of (3-((dimethylamino)methyl)phenyl)boronic acid in step A. 1 H NMR (400MHz, CDCl3) δ7.96(s,1H),7.63(s,1H),7.40–7.30(m,1H),7.01–6.93(m,2H),6.68-6.59(m,1H),6.39(d,J=16.8Hz ,1H),6.26(s,2H),5.80(d,J=10.6Hz,1H),4.66(t,J=7.5Hz,2H),4.04–3.71(m,8H),3.39(s,1H),2.98(s,1H),2.38(s,6H). 19 F NMR (376MHz, CDCl3) δ-111.07, -115.38. LCMS (m / z): 648.5 (M+H).

[0370] Example 5

[0371]

[0372] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-(3-(1-(dimethylamino)ethyl)phenyl)-8-fluoroquinoline-3-carbamate

[0373] The synthesis of Example 5 was carried out in accordance with the method described in Example 3, except that (3-(1-(dimethylamino)ethyl)phenyl)boronic acid was used instead of (3-((dimethylamino)methyl)phenyl)boronic acid in step A. 1H NMR(400MHz,MeOH-d4)δ8.16(d,J=1.6Hz,1H),7.87(s,1H),7.84–7.78(m,1H ),7.61–7.52(m,2H),7.30(dd,J=8.5,5.4Hz,1H),7.09–6.99(m,1H),6.89(d d,J=16.8,10.6Hz,1H),6.31(dd,J=16.7,1.9Hz,1H),5.84(dd,J=10.6,1.9H z,1H),4.12–4.00(m,4H),3.97–3.84(m,4H),2.30(s,6H),1.57–1.45(m,3H). 19 F NMR (376MHz, MeOH-d4) δ -114.82, -118.65 (d, J=9.3Hz). LCMS (m / z): 658.5 (M+H).

[0374] Example 6

[0375]

[0376] 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-(3-((dimethylamino)methyl)phenyl)-8-fluoroquinoline-3-carbamate

[0377] The synthesis of Example 6 was carried out in accordance with the description in Example 3, except that in step A, (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate Int B1) was used instead of 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate Int B2). 1H NMR (400MHz, DMSO-d6) δ8.14(d,J=3.3Hz,1H),7.92(d,J=19.9Hz,2H),7.82(s,1H),7.77(d,J= 7.6Hz,1H),7.56–7.46(m,2H),7.33–7.27(m,1H),7.13–7.06(m,1H),6.94–6.84(m,1H),6.23– 6.14(m,1H),5.78–5.72(m,1H),4.95–4.23(m,4H),4.08(t,J=12.7Hz,1H),4.00–3.90(m,1H), 3.77(t,J=12.7Hz,1H),3.53(s,2H),2.21(s,6H),1.36–1.27(m,3H).LCMS(m / z):658.5(M+H).

[0378] Example 7

[0379]

[0380] 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-(1-(2-(dimethylamino)ethyl)-1H-pyrazol-5-yl)-8-fluoroquinoline-3-carbamate

[0381] The synthesis of Example 6 was carried out in accordance with the description in Example 3. In step A, (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate Int B1) was used instead of 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate Int B2), and (1-(2-(dimethylamino)ethyl)-1H-pyrazol-5-yl)boronic acid was used instead of (3-((dimethylamino)methyl)phenyl)boronic acid. 1H NMR (400MHz, DMSO-d6) δ8.14(s,1H),7.90(d,J=11.8Hz,2H),7.62(d,J=1.9Hz,1H) ,7.29(dd,J=8.6,5.3Hz,1H),7.09(t,J=8.8Hz,1H),6.94–6.83(m,2H),6.17(d,J=1 6.7Hz,1H),5.79–5.70(m,1H),4.62–4.32(m,4H),4.11–3.92(m,3H),3.81–3.66(m ,2H),3.55(s,2H),2.10–1.83(m,6H),1.34–1.20(m,3H).LCMS(m / z):662.5,(M+H).

[0382] Example 8

[0383]

[0384] 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate Step A: (2R)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester

[0385] Add (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate C1, 120 mg, 0.25 mmol), (2-((tert-butoxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boronic acid (232 mg, 0.75 mmol), Pd(dppf)Cl2 (18.3 mg, 25 μmol), and K3PO4 (211 mg, 1.0 mmol) to a microwave tube. After purging with nitrogen three times, add 1,4-dioxane / water (v / v = 3:1, 4 ml), place the tube in a microwave reactor, heat to 90 °C, and react for 1 h. After the reaction was complete, the system was cooled to room temperature, and the reaction solvent was evaporated to dryness using a rotary evaporator. The crude product was purified by FCC (SiO2, EA / PE = 0-80%) to give a yellow solid (2R)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (92 mg, yield 47%). LCMS (m / z): 671.5 (M+H).

[0386] Step B: 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-4-((R)-3-methylpiperazin-1-yl)quinoline-3-carbamate

[0387] Under stirring at room temperature, 2 mL of TFA was added dropwise to a 1 mL solution of (2R)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (92 mg, 0.14 mmol). Stirring continued for one hour, and the reaction was monitored by LCMS to indicate completion. The reaction solvent was evaporated to dryness using a rotary evaporator, and a small amount of DCM was added to concentrate the solution. This process was repeated three times to remove any remaining TFA, yielding crude 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-4-((R)-3-methylpiperazine-1-yl)quinoline-3-carbamate, which was used directly in the next step. LCMS (m / z): 471.3 (M+H).

[0388] Step C: 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate

[0389] Add 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-4-((R)-3-methylpiperazin-1-yl)quinoline-3-carbamate (the crude product obtained in step B) to a round-bottom flask equipped with a magnetic stir bar, and add DCM (3 mL). Cool to 0 °C in an ice-water bath, add DIPEA (2 mL), followed by dropwise addition of a solution of acryloyl chloride (11 mg, 0.12 mmol) in DCM (1.0 mL). After the addition is complete, continue stirring in an ice-water bath for 30 min. After the reaction is complete, add water (5 mL) to quench the reaction, and extract with DCM (10 mL × 3). Combine the organic phases, wash with saturated NaCl (10 mL × 3), and dry with anhydrous Na₂SO₄. After concentration by rotary evaporation, the solution was purified by preparative high performance liquid chromatography to obtain a white solid 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate (26 mg, yield 35%). 1H NMR (400MHz, DMSO-d6) δ8.88 (s, 1H), 8.08 (dd, J = 4.3, 1.4Hz, 1H), 7.92 (s, 2H), 7. 38–7.24(m,1H),7.12–7.04(m,1H),6.86(dd,J=16.6,10.5Hz,1H),6.17(dd,J=16. 6,2.4Hz,1H),5.81–5.68(m,1H),4.47(d,J=125.4Hz,3H),3.94(t,J=12.8Hz,1H), 3.87–3.64(m,2H),3.56–3.41(m,1H),1.49–1.19(m,3H).LCMS(m / z):525.3(M+H).

[0390] Example 9

[0391]

[0392] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate

[0393] The synthesis of Example 9 was carried out in accordance with the description in Example 8, except that in step A, 4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate Int C2) was used instead of (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate Int C1). 1 H NMR (400MHz, Methanol-d4) δ8.78(s,1H),8.10(d,J=1.8Hz,1H),7.26(dd,J=8.4,5.4Hz,1H),7.06–6.97(m,1H),6.87(dd ,J=16.8,10.7Hz,1H),6.30(dd,J=16.8,1.9Hz,1H),5.83(dd,J=10.6,1.9Hz,1H),4.05–3.95(m,4H),3.90–3.78(m,4H). 19 F NMR (376MHz, Methanol-d4) δ-114.73,-119.40. LCMS (m / z): 511.3 (M+H).

[0394] Example 10

[0395]

[0396] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)quinoline-3-carbamate

[0397] Step A: 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0398] In a microwave-safe reaction tube equipped with a magnetic stir bar, add 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate B2, 150 mg, 0.3 mmol), (2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)boric acid (243 mg, 0.9 mmol), and K3PO4 (189 mg, 0.9 mmol) in CH3CN / H2O (v / v = 3:1, 2 mL), and purge with nitrogen for 5 min. Add Pd(dppf)Cl2 (22 mg, 0.03 mmol), and continue purging with nitrogen for 5 min. Microwave heat to 45 °C for 4 hours, then cool to room temperature. Add EA (10 mL) and H2O (5 mL), separate the layers, and extract the aqueous phase with EA (10 mL × 2). The organic phases were combined, washed with saturated NaCl (30 mL), dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by FCC (SiO₂, EA / PE = 0-100%) to give a brown solid, 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (80 mg, yield 44%). LCMS (m / z): 614.4 (M+H).

[0399] Steps B to D: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)quinoline-3-carbamate

[0400] The subsequent synthesis of Example 10 was carried out in accordance with the synthesis described in Example 1, except that in step B, 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester was used instead of (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester. 1H NMR (400MHz, Methanol-d4) δ8.15(s,1H),7.36–7.22(m,4H),7.01(t,J=8.8Hz,1H),6.86(dd,J=16.8,10.6Hz,1H),6.29(dd,J=16.7,1.9Hz,1H),5.82(dd,J =10.5,1.9Hz,1H),4.57(s,2H),3.99(s,4H),3.85(s,4H),3.74(s,2H),2.84( d, J=5.7Hz, 1H), 2.73 (t, J=5.9Hz, 1H), 2.45 (s, 3H). LCMS (m / z): 654.5 (M+H).

[0401] Example 11

[0402]

[0403] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)-1,6-naphthidium-3-carbamate

[0404] The synthesis of Example 11 was carried out in accordance with the description in Example 1, except that in step A, 4-(2,7-dichloro-3-cyano-8-fluoro-1,6-naphthid-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate E1) was used instead of (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate B1). 1 H NMR(400MHz, DMSO-d6)δ9.14(s,1H),7.95(s,2H),7.44(dd,J=8.5,5.8Hz,1H),7.13–7.04(m,1H),6 .91(dd,J=16.6,10.4Hz,1H),6.19(dd,J=16.7,2.4Hz,1H),5.75(dd,J=10.4,2.4Hz,1H),5.47–5.39 (m,1H),4.08–3.99(m,1H),3.89(s,2H),3.83(s,6H),3.34(s,3H),3.08(dd,J=10.0,6.4Hz,1H),2. 98(dd,J=11.1,6.2Hz,1H),2.75(dd,J=11.3,2.8Hz,1H),2.40(dd,J=10.0,4.9Hz,1H),2.29(s,3H). 19F NMR (376MHz, DMSO-d6) δ-111.71, -135.46. LCMS (m / z): 607.5 (M+H).

[0405] Example 12

[0406]

[0407] 4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-2'-amino-6-chloro-5',8-difluoro-[7,8'-bisquinoline]-3-carbonyl

[0408] The synthesis of Example 12 was carried out as described in Example 8, except that in step A, (5-fluoro-2-((4-methoxybenzyl)amino)quinoline-8-yl)boronic acid was used instead of (2-((tert-butoxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boronic acid. LCMS (m / z): 519.1 (M+H).

[0409] Example 13

[0410]

[0411] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-methoxyquinoline-3-carbamate

[0412] The synthesis of Example 13 was carried out in accordance with the description in Example 1, except that in step A, 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate Int B2) was used instead of (R)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate Int B1), and sodium methoxide was used instead of the NaH mixture of (3R,4R)-4-methoxy-1-methylpyrrolidine-3-ol. 1 H NMR (400MHz, DMSO-d6) δ7.94(d,J=5.9Hz,3H),7.25(dd,J=8.4,5.6Hz,1H),7.08(t,J=8.8Hz,1H),6.91(dd,J=16.7,10. 5Hz,1H),6.20(dd,J=16.7,2.2Hz,1H),5.76(dd,J=10.4,2.2Hz,1H),4.05(s,3H),3.87(d,J=15.7Hz,4H),3.68(s,4H). 19 F NMR (376MHz, DMSO) δ-112.27,-119.27. LCMS (m / z): 541.3 (M+H).

[0413] Example 14

[0414]

[0415] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-(1-methyl-1H-pyrazol-5-yl)quinoline-3-carbamate

[0416] The synthesis of Example 14 was carried out in accordance with the description in Example 3, except that (1-methyl-1H-pyrazole-5-yl)boronic acid was used instead of (3-((dimethylamino)methyl)phenyl)boronic acid in step A. 1 H NMR (400MHz, DMSO-d6) δ8.12(s,1H),7.94(s,2H),7.61(s,1H),7.29(dd,J=8.3,5.7Hz,1H),7.10(t,J=8.8Hz,1H), 7.00–6.86(m,2H),6.20(d,J=16.7Hz,1H),5.77(d,J=8.7Hz,1H),4.04(s,3H),3.90(d,J=14.7Hz,4H),3.79(s,4H). 19 FNMR (376MHz, DMSO) δ-112.07,-118.03. LCMS (m / z): 591.4 (M+H).

[0417] Example 15

[0418]

[0419] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-(1,3-dimethyl-1H-pyrazol-4-yl)-8-fluoroquinoline-3-carbamate

[0420] The synthesis of Example 15 was carried out in accordance with the description in Example 3, except that (1,3-dimethyl-1H-pyrazole-4-yl)boronic acid was used instead of (3-((dimethylamino)methyl)phenyl)boronic acid in step A. 1H NMR (400MHz, DMSO-d6) δ8.30(s,1H),8.05(d,J=1.5Hz,1H),7.94(s,2H),7.28(dd,J=8.5,5.6Hz,1H),7.09(t,J=8.8Hz,1H),6.93(dd,J=1 6.7,10.4Hz,1H),6.20(dd,J=16.6,2.4Hz,1H),5.77(dd,J=10.4,2.4Hz,1H),3.89(d,J=11.9Hz,7H),3.73(d,J=5.5Hz,4H),2.41(s,3H). 19 F NMR (376MHz, DMSO) δ-112.23, -118.42. LCMS (m / z): 605.4 (M+H).

[0421] Example 16

[0422]

[0423] 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-4-(4-(2-fluoroacryloyl)piperazin-1-yl)quinoline-3-carbamate

[0424]

[0425] Compound 7-(2-amino-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-8-fluoro-4-(piperazin-1-yl)quinoline-3-carbamate (88 mg, 0.13 mmol), compound 2-fluoroacrylic acid (13.3 mg, 0.15 mmol), and HATU (77 mg, 0.20 mmol) were dissolved in DCM (5 mL). Under stirring at room temperature, DIPEA (52 mg, 0.40 mmol) was added dropwise to the reaction mixture. After the reaction was complete as monitored by LCMS, 20 mL of water was added to the reaction system. The organic phase was separated and collected, concentrated, dried, and purified by preparative high-performance liquid chromatography to obtain a white solid 7-(2-amino-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-8-fluoro-4-(4-(2-fluoroacryloyl)piperazin-1-yl)quinoline-3-carbamate (9 mg, yield 13%). 1 H NMR(400MHz,DMSO-d6)δ8.89(s,1H),8.06(s,1H),7.94(s,2H),7.29(dd,J=8.4,5.6Hz, 1H),7.09(t,J=8.8Hz,1H),5.38–5.33(m,1H),5.32(dd,J=64,4.1Hz,1H),3.91-3.84(m 4H),3.79-3.74(m,4H).19 F NMR(376MHz, DMSO-d6)δ-105.25,-112.15,-117.60.LCMS:529.3(M+H).

[0426] Example 17

[0427]

[0428] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-methylquinoline-3-carbamate

[0429] Step A: 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-methylquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0430] Add 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate B2, 500 mg, 0.99 mmol), methylboronic acid (71 mg, 1.19 mmol), K2CO3 (411 mg, 2.98 mmol), and DME (3 mL) to the reaction flask, and purge with nitrogen for 3 min. Add Pd(dppf)Cl2 (72 mg, 0.099 mmol), continue purging with nitrogen for 3 min, then seal the flask and react at 90 °C for 1 hour. Cool to room temperature. Add EA (10 mL) and H2O (5 mL), separate the layers, and extract the aqueous phase with EA (20 mL × 2). The organic phases were combined, washed with saturated NaCl (30 mL), dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by FCC (SiO₂, EA / PE = 0-100%) to give a yellow solid, 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-methylquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (120 mg, yield 25%). LCMS (m / z): 483.3 (M+H).

[0431] Steps B to D: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-methylquinoline-3-carbamate

[0432] The subsequent synthesis of Example 17 was carried out in accordance with the synthesis described in Example 1, except that in step B, 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-methylquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester was used instead of (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester.1 H NMR (400MHz, DMSO-d6) δ8.04–8.01(m,1H),7.94(s,2H),7.27(dd,J=8.4,5.6Hz,1H),7.12–7.05(m,1H),6.92(dd,J=16. 7,10.5Hz,1H),6.20(dd,J=16.6,2.4Hz,1H),5.76(dd,J=10.5,2.4Hz,1H),3.93–3.82(m,4H),3.69(s,4H),2.74(s,3H). 19 F NMR (376MHz, DMSO-d6) δ-112.20,-118.13. LCMS (m / z): 525.4 (M+H).

[0433] Example 18

[0434]

[0435] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-cyclopropyl-8-fluoroquinoline-3-carbamate

[0436] The synthesis of Example 18 was carried out in accordance with the method described in Example 3, except that cyclopropylboronic acid was used instead of (3-((dimethylamino)methyl)phenyl)boronic acid in step A. 1H NMR (400MHz, DMSO-d6) δ8.01(d,J=1.5Hz,1H),7.96(s,2H),7.27(dd,J=8.5,5.6Hz,1H),7.13–7.05(m,1H),6.94(dd,J=16.7,10.4Hz,1H),6.22(dd ,J=16.6,2.4Hz,1H),5.78(dd,J=10.4,2.4Hz,1H),4.00–3.81(m,4H),3. 79–3.65(m,4H),2.57–2.54(m,1H),1.25–1.12(m,4H).LCMS:551.5(M+H).

[0437] Example 19

[0438]

[0439] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-hydroxyquinoline-3-carbamate

[0440] Step A: 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0441] Under stirring at room temperature, KOH (44 mg, 0.78 mmol) was added to a mixed solution of 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.39 mmol) in THF and H2O (v / v = 2:1, 4 mL). The resulting mixture was stirred at 80 °C for 3 h. The reaction was monitored by LCMS until completion. The reaction solution was concentrated under reduced pressure to remove THF, and then poured into a saturated NH4Cl aqueous solution (4 mL). 1N dilute hydrochloric acid was slowly added dropwise to adjust the pH to 5-6, causing the product to precipitate. The product was filtered, and the filter cake was washed twice with water and petroleum ether, and dried under vacuum to obtain 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (190 mg, crude product), which was directly added to the next step. LCMS(m / z):429.2,431.1(M-56).

[0442] Steps B to D: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-hydroxyquinoline-3-carbamate

[0443] The subsequent synthesis of Example 19 was carried out in accordance with the synthesis described in Example 1, except that in step B, 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester was used instead of (R)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(((3R,4R)-4-methoxy-1-methylpyrrolidine-3-yl)oxy)quinoline-4-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester. 1 H NMR (400MHz, DMSO-d6) δ12.11(s,1H),7.98(s,2H),7.74(s,1H),7.22(dd,J=8.5,5.6Hz,1H),7.11–7.04(m 1H), 6.90 (dd, J=16.6, 10.4Hz, 1H), 6.19 (dd, J=16.7, 2.3Hz, 1H), 5.76 (dd, J=10.4, 2.3Hz, 1H), 3.85 (d, J=19.0Hz, 4H), 3.65 (s, 4H). 19 F NMR (376MHz, DMSO) δ-112.03,-123.05. LCMS (m / z): 527.3 (M+H).

[0444] Example 20

[0445]

[0446] 4-(4-Acryloylpiperazin-1-yl)-2-amino-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate Step A: 4-(7-bromo-6-chloro-3-cyano-2-((diphenylmethylene)amino)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0447] Add 350 mg (694 μmol) of 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester, 126 mg (694 μmol), Pd2(dba)3 (64 mg, 69.4 μmol), XantPhos (80 mg, 139 μmol), Cs2CO3 (679 mg, 2.08 mmol), and dioxane (5 mL) to a microwave tube. After purging with nitrogen for 1 minute, cover the tube and heat it to 80 °C on a microwave-safe surface for 1 hour. After the reaction was complete, the system was cooled to room temperature, and the reaction solution was poured into water (50 mL). Extraction was performed using EA (50 mL × 3), the extract was collected, concentrated, and further purified by FCC (SiO2, EA / PE = 0-50%) to obtain a yellow solid, 4-(7-bromo-6-chloro-3-cyano-2-((diphenylmethylene)amino)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (280 mg, yield 62%). LCMS (m / z): 650.4 (M+H).

[0448] Step B: 4-(7-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-2-((diphenylmethylene)amino)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0449] At room temperature, tert-butyl 4-(7-bromo-6-chloro-3-cyano-2-((diphenylmethylene)amino)-8-fluoroquinoline-4-yl)piperazine-1-carboxylate (280 mg, 431 μmol), (2-((tert-butyloxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boric acid (135 mg, 431 μmol), Pd(dppf)Cl2 (32 mg, 43 μmol), Na2CO3 (137 mg, 1.29 mmol), and a 1,4-dioxane / H2O (v / v = 4:1, 8 mL) solution were added to a reaction tube equipped with a magnetic stir bar. After purging the system with nitrogen for 1 min, the reaction tube was sealed and microwaved to 115 °C for 2 h. Cool to room temperature, dilute with water (60 mL), extract with EA (50 mL × 3), dry under anhydrous Na2SO4, and concentrate under reduced pressure to give 4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-2-((diphenylmethylene)amino)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (310 mg, yield 86%). LCMS (m / z): 836.6 (M+H).

[0450] Step C: 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-((diphenylmethylene)amino)-8-fluoro-4-(piperazin-1-yl)quinoline-3-carbamate

[0451] At room temperature, TFA:CH2Cl2 = 1:1 (15 mL) was added to 4-(7-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-3-cyano-2-((diphenylmethylene)amino)-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (310 mg, 371 μmol). The mixture was stirred for half an hour. After the reaction was detected by LCMS, the mixture was diluted with EA (10 mL) and poured into saturated NaHCO3 (20 mL). The mixture was extracted with EA (30 mL × 3), dried over anhydrous Na2SO4, and the extract was collected to obtain 7-(2-amino-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-2-((diphenylmethylene)amino)-8-fluoro-4-(piperazine-1-yl)quinoline-3-carbamate (140 mg, yield 59%). LCMS(m / z): 636.6(M+H).

[0452] Step D: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-2-((diphenylmethylene)amino)-8-fluoroquinoline-3-carbamate

[0453] Under ice bath conditions, a diluted solution of acryloyl chloride (24 mg, 264 μmol) in CH2Cl2 (0.5 mL) was added dropwise to a mixed solution of 7-(2-amino-7-fluorobenzo[d]thiazolyl)-6-chloro-2-((diphenylmethylene)amino)-8-fluoro-4-(piperazin-1-yl)quinoline-3-carbamate (140 mg, 220 μmol), DIPEA (85 mg, 660 μmol), and CH2Cl2 (5 mL) using a syringe, and stirred for 10 minutes. After the reaction was completed as determined by LCMS, the reaction solution was poured into water (50 mL), extracted with CH2Cl2 (30 mL × 3), the extract was collected, concentrated, and a yellow solid 4-(4-acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-2-((diphenylmethylene)amino)-8-fluoroquinoline-3-carbamate (150 mg, crude product) was obtained. LCMS (m / z): 690.0 (M+H).

[0454] Step E: 4-(4-Acryloylpiperazin-1-yl)-2-amino-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate

[0455] At room temperature, 150 mg (217 μmol) of 4-(4-acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-2-((diphenylmethylene)amino)-8-fluoroquinoline-3-carbamate was dissolved in 30 mL of ethanol, and 1.5 mL of saturated citric acid aqueous solution was added. The mixture was heated to 60 °C and stirred overnight. The product was detected by LCMS. The reaction solution was concentrated, and then water (30 mL) was added to the system. The crude product was concentrated using EA (50 mL × 3) and further separated by preparative high-performance liquid chromatography to obtain 11 mg (10% yield) of 4-(4-acryloylpiperazin-1-yl)-2-amino-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate. 1 H NMR(400MHz,Chloroform-d)δ7.71(d,J=1.6Hz,1H),7.25–7.15(m,1H),7.05–6.91(m,1H),6.69–6.55(m,1H), 6.38(dd,J=16.7,1.8Hz,1H),5.80(dd,J=10.6,1.8Hz,1H),5.68(s,2H),4.06–3.80(m,4H),3.76–3.56(m,4H). 19 F NMR (376MHz, Chloroform-d) δ-111.62, -119.98. LCMS (m / z): 526.4 (M+H).

[0456] Synthesis of compounds 21-2 and 22-2

[0457]

[0458] (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (21-2) and (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (22-2)

[0459] Step A: (S)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (21-1) and (S)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (22-1)

[0460] In a round-bottom flask equipped with a magnetic flask, add (S)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (Int F, 500 mg, 866.2 μmol), Pd(PPh3)4 (100 mg, 87 μmol), formic acid (159 mg, 3.46 mmol), and triethylamine (438 mg, 4.33 mmol). After purging with nitrogen three times, add DMF (15 mL). Heat the reaction system to 55 °C and stir for 6 h. After the reaction was complete, the mixture was cooled to room temperature and poured into 150 mL of water. A yellow solid was produced. The mixture was filtered, and the filter cake was washed with PE (15 mL) and H2O (15 mL). The solid was collected to obtain a mixture (530 mg) of (S)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (21-1) and (S)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (22-1). LCMS (m / z): 544.3 (M+H) (21-1) and LCMS (m / z): 558.3 ​​(M+H) (22-1).

[0461] Step B: (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (21-2) and (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (22-2)

[0462] At room temperature, a mixture (S)-4-(7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (21-1) and (S)-4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (22-1) was added (530m) to a reaction tube equipped with a magnetic stir bar. The reaction mixture consisted of g (976.43 μmol), (2-((tert-butyloxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boronic acid (335.24 mg, 1.07 mmol), Pd(dppf)Cl2 (71.64 mg, 97.64 μmol), Na2CO3 (313 mg, 2.93 mmol), and a 1,4-dioxane / H2O (4:1, 10 mL) solution. After purging with nitrogen for 1 min, the reaction tube was sealed and microwaved to 115 °C for 2 h. The mixture was then cooled to room temperature, diluted with water (60 mL), and extracted with EA (50 mL × 3). The extracts were combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by FCC (SiO2, EA / PE = 0-60%) to give (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (21-2) (80 mg, yield 17%). LCMS (m / z): 730.5 (M+H); and (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (22-2) (380 mg, yield 78%). LCMS (m / z): 746.5 (M+H), 646.4 (M-100+H).

[0463] Example 21

[0464]

[0465] 4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate

[0466] Step A: (4-(6-chloro-3-cyano-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoroquinoline-7-yl)-7-fluorobenzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0467] At room temperature, TMSI (110 mg, 548 μmol) was added dropwise to a mixed solution of (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (80 mg, 110 μmol) and MeCN (3 mL), and the mixture was stirred at room temperature for 5 hours. Then, Et3N (1 mL) was added and stirring was continued for 10 minutes. After the reaction was completed as determined by LCMS, the reaction solution was poured into H2O (30 mL), extracted with EA (30 mL × 3), and the extract was collected to obtain tert-butyl carbamate (68 mg, crude product). LCMS (m / z): 596.4 (M+H).

[0468] Step B: 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoroquinoline-3-carbamate

[0469] At room temperature, TFA:DCM (v / v = 1:1, 5 mL) was added to (4-(6-chloro-3-cyano-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoroquinoline-7-yl)-7-fluorobenzo[d]thiazolyl-2-yl)carbamate tert-butyl ester (68 mg, crude product), and the mixture was stirred at room temperature for half an hour. After the reaction was completed as detected by LCMS, most of the reaction solution was directly concentrated to remove the reaction solution. The reaction solution was then diluted with EA (5 mL) and poured into saturated NaHCO3 (20 mL). The solution was extracted with EA (30 mL × 3), dried over anhydrous Na2SO4, and the extract was collected to obtain 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoroquinoline-3-carbamate (60 mg, crude product). LCMS(m / z):496.3(M+H).

[0470] Step C: 4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate

[0471] Under ice bath conditions, a solution of acryloyl chloride (11 mg, 121 μmol) diluted in DCM (0.5 mL) was added dropwise to a mixture of 7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoroquinoline-3-carbamate (60 mg, crude), saturated NaHCO3 (1.5 mL), and DCM (3 mL) using a syringe, and stirred for 10 minutes. After the reaction was detected by LCMS, the reaction solution was poured into water (30 mL), extracted with DCM (20 mL × 2), the extract was collected and concentrated, and the crude product was further separated by preparative high performance liquid chromatography to obtain a white solid 4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoroquinoline-3-carbamate (7 mg, total yield of three steps 10%). 1 H NMR(400MHz, Methanol-d4)δ8.77(s,1H),8.07(d,J=1.6Hz,1H),7.26–7.11(m,1H),7.00–6.88(m,1H),6.82(s,1H),6.26(dd,J=16.7,1.9Hz,1H ),5.80(dd,J=10.7,1.8Hz,1H),5.24(d,J=24.8Hz,1H),4.21(s,1H),4. 10–3.71(m,4H),3.43(d,J=12.4Hz,1H),3.15(s,1H),3.01–2.86(m,1H). 19 F NMR (376MHz, Methanol-d4) δ-114.73,-118.76. LCMS (m / z): 550.4 (M+H).

[0472] Example 22

[0473]

[0474] 4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-hydroxyquinoline-3-carbamate

[0475]

[0476] Step A: 7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoro-2-hydroxyquinoline-3-carbamate

[0477] At room temperature, TMSI (510 mg, 2.55 mmol) was added dropwise to a mixed solution of (2S)-4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (380 mg, 510 μmol) and MeCN (10 mL), and the mixture was stirred at room temperature for 5 hours. Then, Et3N (3 mL) was added and stirring was continued for 10 minutes. After the reaction was completed as determined by LCMS, the reaction solution was poured into H2O (30 mL), extracted with EA (50 mL × 2), and the extract was collected to give 200 mg of 7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoro-2-hydroxyquinoline-3-carbamate yellow solid (yield 76%). LCMS (m / z): 512.3 (M+H).

[0478] Step B: 4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-hydroxyquinoline-3-carbamate

[0479] Under ice bath conditions, a solution of acryloyl chloride (38 mg, 430 μmol) diluted in CH2Cl2 (0.5 mL) was added dropwise to a mixed solution of 7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-4-((S)-3-(cyanomethyl)piperazin-1-yl)-8-fluoro-2-hydroxyquinoline-3-carbamate (200 mg, 391 μmol), DIPEA (151 mg, 1.17 mmol), and DCM (5 mL) using a syringe, and stirred for 10 minutes. After the reaction was detected by LCMS, the reaction solution was poured into water (50 mL), extracted with DCM (30 mL × 3), the extract was collected and concentrated, and the crude product was further separated by preparative high performance liquid chromatography to obtain a yellow solid 4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-8-fluoro-2-hydroxyquinoline-3-carbamate (52 mg, yield 24%). 1H NMR(400MHz, Methanol-d4)δ7.83(s,1H),7.27–7.16(m,1H),7.04–6.95(m,1H),6.88(s,1H),6.32(dd,J=16.6,1.9Hz,1H ),5.86(dd,J=10.6,1.9Hz,1H),5.48–5.12(m,1H),5.02(s,1H),4.09–3.80(m,4H),3.53–3.39(m,2H),3.05–2.87(m,1H). 19 F NMR (376MHz, DMSO-d6) δ-112.18,-123.14. LCMS (m / z): 566.4 (M+H).

[0480] Examples 23-31

[0481] The following compounds were prepared using a method similar to that described above:

[0482]

[0483]

[0484] Example 32

[0485]

[0486] 4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoro-2-methylquinoline-3-onitrile

[0487] The synthesis of Example 32 was carried out as described in Example 17, except that in step A, (S)-4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate F) was used instead of 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate B2). LCMS (m / z): 564.2 (M+H).

[0488] Example 33

[0489]

[0490] 4-(4-Acryloylpiperazin-1-yl)-2-amino-7-(2-aminobenzo[d]thiazo-4-yl)-6-chloro-8-fluoroquinoline-3-onitrile

[0491] The synthesis of Example 33 was carried out in accordance with the description in Example 20, except that in step B, (2-((tert-butoxycarbonyl)amino)benzo[d]thiazol-4-yl)boronic acid was used instead of (2-((tert-butoxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boronic acid. 1 HNMR(400MHz,DMSO-d6)δ7.80–7.69(m,2H),7.63(s,2H),7.19–7.01(m,4H),6.91(dd,J=16.6,10.5Hz,1H),6.19(d d, J=16.6, 2.4Hz, 1H), 5.75 (dd, J=10.3, 2.4Hz, 1H), 3.97–3.76 (m, 4H), 3.66–3.49 (m, 4H). LCMS (m / z): 508.4 (M+H).

[0492] Example 34

[0493]

[0494] 4-(4-Acryloylpiperazin-1-yl)-2-amino-7-(2-amino-5-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoroquinoline-3-onitrile

[0495] The synthesis of Example 34 was carried out in accordance with the description in Example 20, except that in step B, (2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl)boronic acid was used instead of (2-((tert-butoxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boronic acid. 1 H NMR (400MHz, DMSO-d6) δ8.00–7.67(m,4H),7.24–7.08(m,2H),7.08–6.99(m,1H),6.91(dd,J=16.6,10.5Hz,1H) ,6.19(d,J=16.6Hz,1H),5.76(d,J=10.8Hz,1H),3.99–3.73(m,4H),3.73–3.46(m,4H).LCMS(m / z):526.4(M+H).

[0496] Example 35

[0497]

[0498] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoro-2-(methylamino)quinoline-3-carboxynitrile

[0499] Step A: 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(methylamino)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0500] At room temperature, NaH (143 mg, 3.6 mmol) was added to a mixed solution of methylamine hydrochloride (121 mg, 1.8 mmol) and THF (8 mL) and stirred for 10 minutes. Then, 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (300 mg, 0.60 mmol) was added to the mixture and reacted at room temperature for 2 hours. After LCMS analysis of product formation, the reaction solution was added to H2O (30 mL) and extracted with EA (50 mL × 3). The combined organic phases were washed with saturated brine and dried over anhydrous Na2SO4 to give a yellow solid 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(methylamino)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (200 mg, yield 67%). LCMS (m / z): 499.6 (M+H).

[0501] Steps B to D: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-fluoro-2-(methylamino)quinoline-3-carboxynitrile

[0502] The subsequent synthesis steps of Example 35 were carried out as described in Example 19, except that in step B, 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-(methylamino)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester was used instead of 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester. LCMS (m / z): 540.4 (M+H).

[0503] Example 36

[0504]

[0505] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-2-(dimethylamino)-8-fluoroquinoline-3-onitrile

[0506] The synthesis of Example 36 was performed in accordance with the method described in Example 35, except that dimethylamine hydrochloride was used instead of methylamine hydrochloride in step A. 1H NMR (400MHz, DMSO-d6) δ7.91(s,2H),7.80(s,1H),7.23-7.20(m,1H),7.08–7.03(m,1H),6.94–6.87( m,1H),6.19(d,J=1.6Hz,1H),5.76(d,J=1.2Hz,1H),3.87(d,J=1.6Hz,4H),3.64(s,4H),3.16(s,6H). 19 F NMR (376MHz, DMSO-d6) δ-112.51,-120.42. LCMS (m / z): 554.5 (M+H).

[0507] Example 37

[0508]

[0509] N-(4-(4-acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-2-yl)acetamide

[0510] Step A: 4-(2-acetamido-7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0511] At room temperature, 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (600 mg, 1.2 mmol), acetamide (77 mg, 1.3 mmol), Cs₂CO₃ (1.16 g, 3.6 mmol), and dioxane (10 mL) were added to a round-bottom flask equipped with a magnetic flask. After purging with nitrogen, Pd₂(dba)₃ (109 mg, 0.12 mmol) and Xantphos (65 mg, 0.12 mmol) were added to the system, followed by further purging with nitrogen and heating to 90 °C. The reaction mixture was stirred overnight. The reaction was monitored by LCMS until completion. The reaction solution was then poured into water and extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. This crude product was purified by FCC (SiO2, EA / PE = 0-100%) to give 4-(2-acetamido-7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (205 mg, yield 33%). LCMS (m / z): 526.1 (M+H).

[0512] Steps B to D: N-(4-(4-acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-2-yl)acetamide

[0513] The subsequent synthesis steps of Example 37 were carried out in accordance with the description of Example 19, except that in step B, 4-(2-acetamido-7-bromo-6-chloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester was used instead of 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoro-2-hydroxyquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester. 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.03(d,J=1.5Hz,1H),7.97–7.90(m,2H),7.27(dd,J=8.4,5.6Hz,1H),7.13–7.04(m,1H),6.91(d d,J=16.6,10.4Hz,1H),6.19(dd,J=16.7,2.4Hz,1H),5.76(dd,J=10.4,2.4Hz,1H),3.97–3.81(m,4H),3.74–3.60(m,4H),2.13(s,3H). 19 F NMR (376MHz, DMSO-d6) δ-112.16,-118.37. LCMS (m / z): 568.1 (M+H).

[0514] Example 38

[0515]

[0516] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-2-ethynyl-8-fluoroquinoline-3-nitrile Step A: 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-((trimethylsilyl)ethynyl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0517] At room temperature, 4-(7-bromo-2,6-dichloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (500 mg, 0.99 mmol), ethynyltrimethylsilane (292 mg, 3.0 mmol), Pd(PPh3)4 (115 mg, 0.099 mmol), CuI (38 mg, 0.20 mmol), and TEA (301 mg, 3.0 mmol) were added to DMF (10 mL), purged three times with nitrogen, and reacted at 70 °C for 16 hours. After the product was detected, the reaction solution was added to H2O (100 mL) and extracted with EA (100 mL × 2). The mixture was washed with saturated brine, dried over anhydrous Na2SO4, concentrated, and then subjected to FCC (SiO2, EA / PE = 0-50%) to obtain a yellow solid 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-((trimethylsilyl)ethynyl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (350 mg, yield 62%). LCMS (m / z): 566.0 (M+H).

[0518] Step B: 4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoro-2-((trimethylsilyl)ethynyl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0519] At room temperature, 4-(7-bromo-6-chloro-3-cyano-8-fluoro-2-((trimethylsilyl)ethynyl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (210 mg, 0.37 mmol), (2-((tert-butyloxycarbonyl)amino)-7-fluorobenzothiazol-4-yl)boric acid (139 mg, 0.45 mmol), K₂CO₃ (154 mg, 1.1 mmol), and dioxane / water (v / v = 3:1, 5 mL) were added to a reaction flask equipped with a magnetic stir bar. After purging with nitrogen, Pd(dppf)Cl₂ (27 mg, 0.037 mmol) was added, followed by another nitrogen purging. The mixture was then heated to 90 °C and stirred for 1 hour. After cooling to room temperature, the reaction solution was poured into water and extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. This crude product was purified by FCC (SiO2, EA / PE = 0-100%) to give 4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoro-2-((trimethylsilyl)ethynyl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (90 mg, yield 32%). LCMS (m / z): 753.2 (M+H).

[0520] Step C: 4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-2-ethynyl-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0521] A mixture of 4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-3-cyano-8-fluoro-2-((trimethylsilyl)ethynyl)quinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (90 mg, 0.12 mmol), K₂CO₃ (33 mg, 0.24 mmol), and acetonitrile (2 mL) was stirred at room temperature for 1 h. The reaction was monitored by LCMS until completion, and the mixture was poured into water and extracted with EA. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 4-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-6-chloro-3-cyano-2-ethynyl-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (70 mg, crude), which was used directly in the next reaction without further purification. LCMS(m / z): 681.2(M+H).

[0522] Steps D and E: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-2-ethynyl-8-fluoroquinoline-3-nitrile

[0523] The steps for protecting group removal and allyl chloride introduction in the subsequent synthesis of Example 38 can be basically carried out in accordance with the method described in Example 1. LCMS (m / z): 535.1 (M+H).

[0524] Example 39

[0525]

[0526] 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-methoxyquinoline-3-nitrile Step A: 4-(7-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-methoxyquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0527] Add 70 mg (0.11 mmol) of 4-(7-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-fluoroquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester and 20 mg (0.37 mmol) to a reaction flask equipped with a magnetic flask. Molecular sieve (100 mg) and dioxane (2 mL) were added, the bottle cap was tightened, and the mixture was heated to 110 °C and stirred for 8 h. After the reaction was complete, EA (20 mL) was added to dilute the reaction system, and the mixture was washed successively with water (15 mL) and saturated brine (15 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, concentrated under reduced pressure, and the crude product was purified by FCC (SiO2, EA / PE = 0-80%) to obtain 4-(7-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazolyl-4-yl)-6-chloro-3-cyano-8-methoxyquinoline-4-yl)piperazine-1-carboxylic acid tert-butyl ester (42 mg, yield 59%). LCMS (m / z): 669.1 (M+H).

[0528] Steps B and C: 4-(4-Acryloylpiperazin-1-yl)-7-(2-amino-7-fluorobenzo[d]thiazo-4-yl)-6-chloro-8-methoxyquinoline-3-nitrile

[0529] The steps for removing the protecting group and introducing allyl chloride involved in the subsequent synthesis of Example 39 can be basically carried out with reference to the method described in Example 1. 1 H NMR(400MHz, DMSO-d6)δ8.86(s,1H),7.96(s,1H),7.89–7.82(m,2H),7.16(dd,J=8.4,5.7Hz,1H),7.07–7.01(m,1H),6.91(dd,J= 16.7,10.5Hz,1H),6.19(dd,J=16.7,2.4Hz,1H),5.75(dd,J=10.4,2.4Hz,1H),3.94–3.82(m,4H),3.78(s,3H),3.70–3.64(m,4H). 19 F NMR(376MHz,DMSO-d6)δ-113.34.LCMS(m / z):523.1(M+H).

[0530] Active Examples

[0531] Example 1: Inhibitory effect of the compound of the present invention on the proliferation of KRas G12C mutant cells

[0532] This experiment uses Promega's... The Luminescent Cell Viability Assay kit was used to evaluate and validate the inhibitory activity of the compounds of this invention against the proliferation of KRas G12C-mutant NCI-H358 human non-small cell lung cancer cells. [Experimental Materials]: NCI-H358 cell line (Cell Resource Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Resource No.: 3111C0001CCC000470), 96-well clear flat-bottomed black-walled cell culture plates (Greiner BioOne, Catalog No. #655096), RPMI-1640 medium (GE, Catalog No. #SH30809.01), fetal bovine serum (FBS) (Thermo Fisher Scientific, Catalog No. #10099-141). Luminescent Cell Viability Assay Kit (Promega, catalog number #G7573), PBS (Solarbio, catalog number #P1020), trypsin (Thermo Fisher, catalog number #25200072), DMSO (Sigma, catalog number #D2650), and Methylcellulose (SIGMA, catalog number #9004-67-5).

[0533] [Experimental Procedure]: Add 180 μL of cell suspension (RPMI 1640 solution containing 1% methylcellulose and 10% FBS) to a 96-well cell culture plate to achieve a cell density of 1500 viable cells / well. Set up a control group (culture medium control) containing only 3D complete culture medium (RPMI 1640 solution containing 1% methylcellulose and 10% FBS) without cells or the compound, and a control group (cell control) containing cells without the compound. During the assay, use compound AMG510 or a lower reference compound as a positive control. Incubate the cell culture plate overnight in a cell culture incubator. Prepare a 10-fold drug solution (RPMI 1640 solution containing 1% DMSO and 10% FBS) at a concentration of 10 μM. Add 20 μL of the drug solution to each well of a 96-well plate seeded with cells, bringing the final concentration of the compound in each well to 1 μM. Set up three replicates for each compound. The DMSO content is 0.1%. Prepare and add a solution of compound AMG510 or a lower reference compound to the positive control wells in the same manner. Incubate the cell culture plates in a cell culture incubator for 120 h. For endpoint detection, melt the CellTiter-Glo reagent and allow the cell culture plates to equilibrate to room temperature for 30 min. Add 100 μL of CellTiter-Glo to each well of the cell culture plate and shake on a track-mounted shaker for 5 min to fully lyse the cells. Place the cell culture plates at room temperature for 20 min to stabilize the bioluminescence signal. Scan the bioluminescence values ​​of each well using a multi-mode microplate reader (Molecular Devices, Spectramax M3 microplate reader) at all wavelengths.

[0534] [Test Samples] Compounds from Examples 1-39, and reference compound A (Prepared and characterized according to the method described in WO2020 / 081282A1) and reference compound B (Prepared and characterized according to the method described in WO2015054572).

[0535] [Data Analysis] The cell inhibition rate under the action of each compound was calculated using the following formulas and GraphPad Prism 7.0 software:

[0536] Inhibition rate % = [1-(Lum)] 待测药 -Lum 培养液对照 ) / (Lum 细胞对照 -Lum 培养液对照 )】×100%

[0537] IC 50 The dose-response curve was obtained by using GraphPad Prism 7.0 software and nonlinear S-curve regression to fit the data, and then the values ​​were calculated.

[0538] [Experimental Results] The compounds of this invention exhibited satisfactory anti-proliferative activity against KRas G12C-mutant NCI-H358 human non-small cell lung cancer cells. Specifically, all the compounds tested in the examples showed anti-proliferative activity, with an IC50 value of [missing value]. 50 The values ​​are generally <1 μM, for example <0.5 μM, <0.1 μM, preferably <50 nM, more preferably <20 nM, and most preferably <10 nM. For example, compounds in Examples 17, 20, 32, 33, 34, and 37 all showed IC50 values ​​<50 nM. 50 Values, compounds in Examples 1, 2, 3, 5, 6, 8, 9, 10, and 39 showed IC50 values ​​<20 nM. 50 Values. Specific data for some representative example compounds are shown in Table 1.

[0539] Table 1. Inhibitory activity of compounds from representative examples on NCI-H358 cell proliferation (inhibition rate at 1 μM concentration and IC50) 50 )

[0540] Example 1 94.7 0.015 Example 2 97.6 0.003 Example 3 97.7 0.009 Example 4 94.8 0.053 Example 5 94.9 0.007 Example 6 95.0 0.017 Example 7 90.0 0.26 Example 8 94.0 0.018 Example 9 98.8 0.0086 Example 10 96.1 0.018 Reference compound A 96.9 0.011 Reference compound B / 0.35

[0541] Example 2: Pharmacokinetic characteristics of the compounds of the present invention in rats

[0542] 2.1 The pharmacokinetic characteristics of some compounds of the present invention were evaluated by a rat cassette pharmacokinetic experiment. [Experimental Materials]: Male SD rats, 6-8 weeks old, weighing 220-250g, purchased from Zhaoyan (Suzhou) New Drug Research Center Co., Ltd.; Tolbutamide (Aladdin, catalog number H1401054); Sulfobutyl β-cyclodextrin (Captisol, Shandong Binzhou Zhiyuan Biotechnology, catalog number 20191013); Propylene glycol (15) stearate (Solutol, Meilun Biotechnology, catalog number S0206A); DMSO (Vetec, catalog number WXBD0293V); Acetonitrile (Sigma-Aldrich, catalog number WXBD1744V); Methanol (Sigma-Aldrich, catalog number WXBD2831V).

[0543] [Experimental Procedure]: The compounds were prepared in a solvent of 5% DMSO / 10% Solutol / 85% (20% Captisol) to achieve a final concentration of 1 mg / mL for each compound. The drug formulation was injected into SD rats via the tail vein at an injection volume of 1 mL / kg. Blood was collected from the external jugular vein at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h. The plasma was collected by centrifugation at low temperature for 20 minutes and stored at -80℃ for analysis.

[0544] [Sample Analysis]: Establish LC-MS / MS analytical methods for compounds.

[0545] Preparation of standard curves: For each compound, take 20 μL of 1 mg / mL DMSO stock solution and transfer it to 900 μL of 50% methanol working solution. Dilute stepwise to obtain a standard curve working solution with concentrations of 20000, 10000, 5000, 1000, 500, 100, 50, 20, and 10 ng / mL. Then, take 5 μL of the standard curve working solution and mix it with 45 μL of rat blank plasma to obtain a standard curve with concentrations of 2000, 1000, 500, 100, 50, 10, 5, 2, and 1 ng / mL, which is used for quantification of unknown samples.

[0546] Sample pretreatment: Add 250 μL of acetonitrile containing internal standard as a precipitant to 50 μL of unknown plasma sample and standard curve sample to precipitate plasma proteins, extract the test compound from the plasma, centrifuge at low temperature for 20 minutes, take the supernatant, mix the supernatant with 0.1% formic acid aqueous solution, and inject 5 μL to analyze the drug blood concentration.

[0547] [Data Processing]: Standard curves were plotted using mass spectrometry analysis software, unknown samples were quantified, and pharmacokinetic parameters were calculated using Winnonlin 8.2 based on the drug concentrations of the unknown samples at each time point.

[0548] [Experimental Results]: The experimental results show that, in the cassette-based pharmacokinetic evaluation, the compounds of this invention exhibit good or even improved pharmacokinetic properties.

[0549] Table 2.

[0550]

[0551]

[0552] Compound AMG510: Prepared according to the method described in Lanman B et al., J. Med. Chem. 2020, 63, 52-65.

[0553] 2.2 Pharmacokinetic properties of representative compounds of this invention

[0554] [Experimental Materials] Same as 2.1 above.

[0555] [Experimental Procedure]: Rats were administered the drug intravenously using a method similar to that described in 2.1 above, with a compound concentration of 3 mg / mL. Drug formulation, administration, and sample collection were performed as described in 2.1. For the oral administration group, the drug was prepared as a 3 mg / mL suspension of 0.5% methylcellulose MC-400cp (Aladdin, M112866) and administered by gavage at a concentration of 10 mL / kg. Blood samples were collected from the external jugular vein at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. The plasma was collected by centrifugation at low temperature for 20 minutes and stored at -80℃ for analysis.

[0556] Subsequent sample analysis and data processing methods are similar to those described in 2.1 and should be performed accordingly.

[0557] [Experimental Results]: The results show that Example 9 has excellent pharmacokinetic properties, as detailed in Table 3.

[0558] Table 3

[0559]

[0560] Example 3: Antitumor activity of the compounds of the present invention in a human non-small cell lung cancer NCI-H358 xenograft mouse model.

[0561] This experiment evaluated and verified the proliferation inhibitory activity of the compounds of this invention in a human non-small cell lung cancer NCI-H358 xenograft mouse model.

[0562] [Experimental Materials]: The NCI-H358 cell line carrying the KRAS G12C mutation was provided by Kangyuan Biotech (Beijing) Co., Ltd. (derived from ATCC, catalog number CRL-5807). Female NPSG mice were provided by Beijing Finoc Biotechnology Co., Ltd.

[0563] [Experimental Procedure]: 6-8 week old female NPSG mice were subcutaneously inoculated with 5×10⁵ lbsin injections in the right shoulder area. 6 One NCI-H358 cell (containing 50% matrix gel) was seeded at a volume of 0.1 mL. When the tumor grew to an average volume of 160–220 mm... 3 Mice were randomly assigned to groups based on tumor size and body weight for drug administration. Drug administration began immediately after grouping, with the day of administration considered day 0. Administered once daily by gavage at a dose of 10 mg / kg or a solvent control (50 mM citrate buffer containing 10% cyclodextrin, pH 5.0). Tumor volume and body weight were measured twice weekly during the experiment. Tumor volume was calculated using the formula V = D × d × d / 2, where D is the long axis of the tumor and d is the short axis. The mean tumor inhibition rate (TGI%) was calculated as follows: [(C...] 平均值 -C 0平均值 )-(T 平均值 -T 0平均值)] / (C 平均值 -C 0平均值 )*100%, where T is the tumor volume of the treatment group, T0 is the initial tumor volume of the treatment group, C is the tumor volume of the control group, and C0 is the initial tumor volume of the control group.

[0564] [Experimental Results]:

[0565] The experimental results, shown in Table 4, indicate that the representative compounds of this invention significantly inhibited the growth of NCI-H358 tumors. Under the same dosage conditions, the compounds of this invention showed comparable or superior tumor-suppressing effects compared to the control compound AMG510. Meanwhile, there were no significant changes in the body weight of mice in any group.

[0566] Table 4

[0567] Solvent control group 1170 / Example 1 478.4 71 Example 2 497.3 69 Example 8 519.2 67 AMG510 525.1 66

[0568] In addition, using the same materials and methods as described above, the tumor-suppressive activity of the compound of Example 9 at different doses in the aforementioned non-small cell lung cancer mouse model, as well as its effect on body weight, were further investigated. The results are attached. Figure 1 As shown, the compounds in the representative examples exhibit excellent tumor-suppressive activity and have no significant side effects on body weight.

[0569] Example 4: Inhibition test of the compound of the present invention on cytochrome P450

[0570] This experiment evaluates the inhibitory effect of the invented compound on cytochrome P450.

[0571] [Experimental Materials]: Human liver microsomes (Corning, catalog number 452161); reduced nicotinamide adenine dinucleotide phosphate (NADPH, MCE, catalog number HY-F0003 / CS-4998); phenacetin, diclofenac, α-naphthylflavonoid, omeprazole, and ketoconazole were purchased from TCI; S-mpheniramine and testosterone were purchased from CAYMAN; midazolam was purchased from Bioreclamation IVT; quinidine was purchased from Damas-beta; sulfamethoxazole was purchased from MCE; and bufrolol was purchased from TRC. [Experimental Procedure]:

[0572] Prepare 0.1M potassium phosphate buffer (K-buffer): Prepare 100mM potassium phosphate buffer (K-buffer) using potassium dihydrogen phosphate and dipotassium hydrogen phosphate, and adjust the pH to 7.4.

[0573] Preparation of 400× test compound and reference inhibitor: Dissolve 8 μL of 10 mM test compound stock solution in 12 μL acetonitrile. Preparation of mixed solutions of CYP1A2, CYP2C9 and CYP2D6 inhibitors: Mix 12 μL of 1 mM α-naphthylflavonoid, 10 μL of 40 mM sulfadiazine, 10 μL of 10 mM quinidine and 8 μL of DMSO solution. Preparation of inhibitor solutions of CYP3A4 and CYP2C19: Dissolve 8 μL of DMSO solution in 12 μL acetonitrile.

[0574] Prepare 4× NADPH potassium phosphate solution: Add 66.7 mg NADPH to 10 mL of 0.1 M K-buffer, pH 7.4. Prepare 4× substrate potassium phosphate solution: Prepare solutions of different substrates to the required concentrations (4 times the concentration) using 10 mL of 0.1 M K-buffer.

[0575] Prepare a 0.2 mg / mL human liver microsome (HLM) solution: Add 10 μL of 20 mg / mL human liver microsomes to 990 μL of K-buffer and store on ice until use.

[0576] Add 600 μL of 0.2 mg / mL HLM to a 96-well plate, followed by 3 μL of a 400-fold dilution of the test compound solution; add 200 μL of 0.2 mg / mL HLM to the same well, followed by 1 μL of diluted positive control inhibitor solution. Aliquot 30 μL of the compound-human liver microsome mixture into the 96-well plate, and then add 15 μL of the substrate solution. Preheat the obtained solutions and prepared NADPH solution at 37 °C for 5 min. Add 15 μL of the preheated NADPH solution to the reaction plate, mix well, and begin the reaction. Incubate the reaction plate at 37 °C. React 3A4 for 5 min; 1A2, 2C9, 2D6 for 10 min; and 2C19 for 45 min. At the end of the reaction, terminate the reaction by adding 120 μL of acetonitrile containing the internal standard. Vortex the sample for 10 min, centrifuge at 5594 g for 15 min, and then analyze the prepared sample by LC-MS / MS.

[0577] [Experimental Results]:

[0578] The experimental results (Table 5) show that, at the tested concentrations, compared with AMG510 and reference compound A, the representative compounds of this invention did not significantly inhibit the key CYP subtypes of drug metabolism, demonstrating better drug-drug interaction safety.

[0579] Table 5

[0580]

[0581] Example 5: Inhibitory effect of the compound of the present invention on the proliferation of a series of KRas mutant cells

[0582] This experiment used the CellTiter-Glo (CTG) kit from Promega to evaluate the antiproliferative activity of the representative compounds of this invention against 12 KRas mutant tumor cell lines.

[0583]

Experimental Materials

[0584] [Experimental Method]:

[0585]

[0586] Each of the above cell lines was cultured separately in the complete culture medium shown at 37°C and 5% CO2. Cells in the logarithmic growth phase were harvested and counted using a platelet counter. Cell viability was assessed using the trypan blue rejection assay to ensure it was above 90%. Cell density was adjusted using complete culture medium, and then 90 μL of the mixture was seeded into 96-well cell culture plates, for a total of 3000 cells. The cells in the 96-well plates were incubated at 37°C and 5% CO2.

[0587] Prepare 10-fold culture medium solutions of the test compounds, with a maximum detection concentration of 10 μM. Nine concentrations were prepared and diluted 3.16-fold. Then, 10 μL of each diluted compound solution was transferred to the corresponding wells of a 96-well cell culture plate, with three replicates per concentration. Cells in the drug-treated 96-well plates were cultured at 37°C and 5% CO2 for 72 hours, followed by CTG analysis.

[0588] Melt the CTG reagent and equilibrate the cell plate to room temperature for 30 minutes. Add 100 μL CTG solution to each well. Initiate cell lysis by shaking on a track-mounted shaker for 5 minutes. Place the cell plate at room temperature for 20 minutes to stabilize the cryo-light signal, read the cryo-light values, and collect data. Analyze the data using GraphPad Prism 7.0 software. Utilize nonlinear S-curve regression to fit the data to derive the dose-response curve, and calculate the relative and absolute IC50 values ​​using methods well-known to those skilled in the art. 50 Value, and maximum inhibition rate %.

[0589] Inhibition rate % = [1-(Lum)] 待测药 -Lum 培养液对照 ) / (Lum 细胞对照 -Lum 培养液对照 )】×100%

[0590] [Experimental Results]:

[0591] Table 6.

[0592]

[0593]

[0594] Table 6 shows the experimental results, which indicate that the representative compound of this invention exhibits superior antiproliferative activity compared to reference compound A in a range of KRas tumor cells, and also demonstrates good selectivity.

[0595] Example 6: Inhibitory effect of the compound of the present invention on the proliferation of cells in a KRas G12C inhibitor-resistant model.

[0596] Acquired resistance is one of the key factors affecting the therapeutic effect of KRas G12C inhibitors. This experiment evaluates the potential application value of the compound of the present invention in a known resistance model by measuring the acquired resistance of the KRas G12C inhibitor Adagrasib (Engl J Med 2021; 384:2382-93.).

[0597] This experiment used the CellTiter-Glo (CTG) kit provided by Promega to evaluate the effect of the test compounds on the cell proliferation of five KRas cell lines (which have acquired resistance to the KRas G12C inhibitor Adagrasib), with reference compound A and AMG510 as control compounds.

[0598] [Experimental Materials]: The experimental materials and equipment used are the same as those listed in Example 5 above. All cell lines used in this experiment were obtained from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd.

[0599] [Experimental Method]:

[0600]

[0601] The inhibitory activity of the test compounds on cell growth was investigated using the same methods and conditions as in Example 5, and the results are shown in Table 7.

[0602] Table 7.

[0603]

[0604]

[0605] The results showed that the representative compound of the present invention exhibited superior inhibitory activity compared to reference compound A and AMG510 in the above-mentioned drug-resistant cell model.

[0606] Example 7: In vivo pharmacodynamic study of the compounds of the present invention against a BALB / c nude mouse model of subcutaneous xenografted human pancreatic cancer Mia PaCa-2 cells.

[0607] The in vivo efficacy of the representative compound of this invention in a human pancreatic cancer Mia PaCa-2 cell subcutaneous xenograft tumor model was evaluated.

[0608] [Experimental Materials]: BALB / c nude mice, 6-8 weeks old, female, from Zhejiang Vital River Laboratory Animal Technology Co., Ltd. Human pancreatic cancer Mia PaCa-2 cells (ATCC, catalog number: CRL-1420). Matrigel (Matrigel, Corning, catalog number 356234).

[0609] [Experimental Method]: 0.2 mL (5 × 10⁻⁶) 6 Mia PaCa-2 cells (with matrix gel, volume ratio 1:1) were subcutaneously inoculated into the right posterior dorsal region of each mouse, resulting in an average tumor volume of approximately 137 mm². 3 Mice were randomly assigned to experimental groups (solvent control group; AMG510 10 mpk; compound 9 of Example 9 3 mpk; compound 9 of Example 9 10 mpk; compound 9 of Example 9 30 mpk), with 8 mice in each group. Drug administration was initiated once daily by gavage to a 0.5% methylcellulose (400 cp) aqueous solution. Tumor volume and body weight were measured twice weekly during the experiment.

[0610] The tumor volume is calculated using the following formula: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively.

[0611] The tumor inhibition efficacy (TGI) of the test compound was calculated according to the following formula: TGI (%) = [1 - (mean tumor volume at the end of treatment - mean tumor volume at the beginning of treatment) / (mean tumor volume at the end of treatment in solvent control group - mean tumor volume at the beginning of treatment in solvent control group)] × 100%.

[0612] [Experimental Results]: See Table 8 and appendix for experimental results. Figure 2 The results showed that the representative compound exhibited excellent target-related tumor suppression activity in human pancreatic cancer Mia PaCa-2, which was superior to the control compound AMG510, and there was no significant change in body weight in any of the groups of animals.

[0613] Table 8.

[0614] Solvent control group 1202 / Example 9 (30mpk) 162 97.7％ Example 9 (10mpk) 246 89.8％ Example 9 (3mpk) 665 50.4％ AMG510 (10mpk) 432 72.3％ .

Claims

1. Compounds of formula (I), (I) in, A is selected from CR a , where R a Selected from halogens; R 1 R 2 and R 3 Each is independently selected from H; R b Each time it appears, it is independently selected from H; -X-R4 is selected from -OC 1-6 Alkyl groups, and -O- A 4-6 membered heterocyclic alkyl group comprising 1, 2, or 3 heteroatoms independently selected from N, O, or S, wherein the heterocyclic alkyl group is composed of 1, 2, or 3 heteroatoms independently selected from C. 1-6 Alkyl and -(CR) c R c ) 0-6 -OR c Substitution of groups, R c Selected from H or C 1-6 alkyl; E is selected from halogens; R 5 for ; R 6 For halogens, R 7 and R 8 Each is independently represented by H; Or its isomers or pharmaceutically acceptable salts.

2. The compound of formula (I) according to claim 1, or an isomer thereof or a pharmaceutically acceptable salt thereof, wherein A is a CR a , where R a It is Cl.

3. The compound of formula (I) according to any one of claims 1-2, or an isomer thereof or a pharmaceutically acceptable salt thereof, wherein -X-R4 is -OC. 1-6 alkyl.

4. The compound of formula (I) according to claim 3, or an isomer thereof or a pharmaceutically acceptable salt thereof, wherein -X-R4 is -O-CH3.

5. The compound of formula (I) according to any one of claims 1-2, or an isomer thereof or a pharmaceutically acceptable salt thereof, wherein -X-R4 is a -O- 4-6 membered heterocyclic alkyl group containing one N heteroatom, wherein the heterocyclic alkyl group is C 1-6 Alkyl and -OR c Replace, R c -C 1-6 alkyl.

6. The compound of formula (I) according to claim 5, or an isomer thereof or a pharmaceutically acceptable salt thereof, wherein R4 in -X-R4 is .

7. The compound of formula (I) according to any one of claims 1-2, or an isomer thereof or a pharmaceutically acceptable salt thereof, wherein E is F.

8. The compound of formula (I) according to any one of claims 1-2, or an isomer thereof or a pharmaceutically acceptable salt thereof, wherein R 5 for .

9. A pharmaceutical composition comprising a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof, according to any one of claims 1-8, and a pharmaceutically acceptable excipient.

10. Use of a compound, isomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of claims 1-8, in the preparation of a medicament for the prevention or treatment of diseases mediated by KRas G12C mutations, wherein the diseases are selected from lung adenocarcinoma, lung cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, and esophageal cancer.

11. The use according to claim 10, wherein the disease mediated by the KRas G12C mutation is selected from lung adenocarcinoma, colon cancer, rectal cancer, pancreatic cancer, and bile duct cancer.

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