Dihydrofuro[3,4-f]quinazoline compound, preparation method therefor and use thereof in medicine

Abstract

The present disclosure relates to a dihydrofuro[3,4-f]quinazoline compound, a preparation method therefor and a use thereof in medicine. Specifically, the present disclosure relates to a dihydrofuro[3,4-f]quinazoline compound represented by formula (1), a preparation method therefor, a pharmaceutical composition containing the compound, and a use of the compound as a therapeutic agent, particularly a use of the compound in preparation of a drug for inhibiting KRAS amplification and / or mutant activity.

Description

Dihydrofuro[3,4-f]quinazoline compounds, preparation methods thereof and application thereof in medicine TECHNICAL FIELD

[0001] The present disclosure belongs to the field of medicine, and relates to a dihydrofuro[3,4-f]quinazoline compound, a preparation method thereof and application thereof in medicine. In particular, the present disclosure relates to a dihydrofuro[3,4-f]quinazoline compound shown in Formula 1, a preparation method thereof, a pharmaceutical composition containing the compound, and the use thereof in the preparation of a medicament for inhibiting KRAS amplification and / or mutant activity. BACKGROUND

[0002] KRAS protein encoded by KARS gene is a small GTPase belonging to the RAS superfamily of proteins. In cells, KRAS protein is transformed between inactivation and activation, and is in an inactivated state when KRAS binds to guanosine diphosphate (GDP), and is in an activated state when KRAS binds to guanosine triphosphate (GTP), causing activation of downstream signaling pathways. KRAS can be activated by upstream tyrosine kinases such as EGFR, and the downstream pathways activated by KRAS are commonly the RAS-RAF-MEK-ERK and PI3K-AKT-mTOR signaling pathways that regulate cell proliferation and growth.

[0003] KRAS is one of the most common oncogenes in solid tumors, and about 19% of tumors have KRAS mutations, including ~90% of pancreatic cancer, ~50% of colon cancer, ~30% of lung adenocarcinoma, etc. There is also a certain proportion in other cancer types such as cholangiocarcinoma, cervical cancer, bladder cancer, liver cancer and breast cancer, etc. The most common sites of genetic mutations are codons 12, 13 and 61, of which the mutation of codon 12 is the most common. KRAS mutations cause RAS to be more in the activated state of GTP binding, activating downstream pathways. In addition, KRAS amplification / overexpression or upstream activation also occurs in tumors, which all cause persistent activation of RAS downstream pathways, leading to tumor occurrence.

[0004] Due to the lack of traditional small molecule binding sites on the surface of KRAS protein and the extremely high affinity for guanylate, it is difficult to be competitively inhibited by small molecules, and KRAS has been considered as an undruggable drug target for a long time. However, due to the importance and universality of KRAS abnormal activation in cancer progression, KRAS has always been and still is a target of great concern for drug development. Currently, only inhibitors targeting KRAS G12C and G12D have been approved or are in clinical phase, and therefore there is still a need to develop pan KRAS inhibitors for the treatment of various KRAS mutant tumors or KRAS-dependent tumors.

[0005] The currently published related patent applications include WO2023183585A1, etc. SUMMARY

[0006] Typical compounds of the present disclosure include, but are not limited to, those shown in Table A:

[0007] Typical intermediate compounds of the present disclosure include, but are not limited to, those shown in Table B:

[0008] Another aspect of the present disclosure relates to a pharmaceutical composition containing a compound shown in Table A of the present disclosure, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0009] The present disclosure further relates to the use of a compound shown in Table A, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, in the manufacture of a medicament for inhibiting KRAS amplification and / or KRAS mutant activity; in some embodiments the KRAS mutant is selected from one or more of KRAS G12A, G12C, G12D, G12V, G12R, G12S, G13A, G13C, G13D, G13R, G13S, G13V, Q61E, Q61H, Q61K, Q61L, Q61P, Q61R, A146T, A146P, A146V, and A146T mutations; in some embodiments KRAS G12D and / or KRAS G12V mutations.

[0010] The present disclosure further relates to the use of a compound shown in Table A, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, in the manufacture of a medicament for treating and / or preventing a disease or disorder mediated by KRAS amplification and / or KRAS mutant.

[0011] The present disclosure further relates to the use of a compound shown in Table A, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, in the manufacture of a medicament for treating and / or preventing a cancer; in some embodiments the cancer is selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pleural cancer, peritoneal cancer, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial cancer, urethral cancer, bladder cancer, anal cancer, joint cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, melanoma, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma, and glioblastoma; in some embodiments, from pancreatic cancer, colorectal cancer, and non-small cell lung cancer.

[0012] The present disclosure further relates to a method of inhibiting the activity of KRAS amplifications and / or KRAS mutant forms comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Table A or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0013] The present disclosure further relates to a method of treating and / or preventing a disease or disorder mediated by KRAS amplifications and / or KRAS mutant forms comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Table A or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0014] The present disclosure further relates to a method of treating and / or preventing cancer comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Table A or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0015] The present disclosure further relates to a compound of Table A or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use as a medicament.

[0016] The present disclosure further relates to a compound of Table A or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in inhibiting the activity of KRAS amplifications and / or KRAS mutant forms.

[0017] The present disclosure further relates to a compound of Table A or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in treating and / or preventing a disease or disorder mediated by KRAS amplifications and / or KRAS mutant forms.

[0018] The present disclosure further relates to a compound of Table A or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in treating and / or preventing cancer.

[0019] The disease or disorder referred to in the present disclosure is a disease or disorder that is treated and / or prevented by inhibiting the activity of KRAS amplifications and / or KRAS mutant forms.

[0020] The disease or disorder mediated by KRAS amplification and / or KRAS mutant described in the present disclosure is cancer, in some embodiments, the KRAS mutant is selected from one or more of KRAS G12A, G12C, G12D, G12V, G12R, G12S, G13A, G13C, G13D, G13R, G13S, G13V, Q61E, Q61H, Q61K, Q61L, Q61P, Q61R, A146T, A146P, A146V, and A146T mutations, in some embodiments, KRAS G12D and / or KRAS G12V mutations; in some embodiments, the cancer is selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, throat cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pleural cancer, peritoneal cancer, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial cancer, urethral cancer, bladder cancer, anal cancer, joint cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, melanoma, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma, and glioblastoma; in some embodiments, pancreatic cancer, colorectal cancer, and non-small cell lung cancer.

[0021] The colorectal cancer described in the present disclosure is colon cancer or rectal cancer.

[0022] The brain cancer described in the present disclosure is selected from glioblastoma multiforme or neuroblastoma; the soft tissue cancer is selected from fibrosarcoma, gastrointestinal stromal tumor, rhabdomyoma, leiomyosarcoma, dedifferentiated liposarcoma, pleomorphic liposarcoma, malignant fibrous histiocytoma, round cell sarcoma, and synovial sarcoma; the lymphoma is selected from Hodgkin's disease and non-Hodgkin's lymphoma (e.g., mantle cell lymphoma, diffuse large B-cell lymphoma, follicular center lymphoma, marginal zone B-cell lymphoma, lymphoplasmacytic lymphoma, and peripheral T-cell lymphoma); in some embodiments, the liver cancer is hepatocellular carcinoma; the lung cancer (also known as bronchogenic lung cancer) is selected from non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), and squamous cell carcinoma; the kidney cancer is selected from renal cell carcinoma, clear cell and renal oncocytoma; the leukemia is selected from chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia, acute lymphoblastic leukemia (ALL), T-cell acute lymphoblastic leukemia (T-ALL), chronic myelogenous leukemia (CML), and acute myeloid leukemia (AML); the skin cancer is selected from malignant melanoma, squamous cell carcinoma, basal cell carcinoma, and angiosarcoma; in some embodiments, the myeloma is multiple myeloma.

[0023] The active compounds can be prepared into various pharmaceutically acceptable dosage forms for administration by any of the routes conventionally used for such drugs, using routine methods familiar to pharmaceutical chemists. Thus, for example, the active compounds of the present disclosure can be prepared as various dosage forms for oral, injectable (e.g., intravenous, intramuscular, or subcutaneous), inhalant, or insufflation administration. The compounds of the present disclosure can also be formulated into dosage forms such as tablets, hard or soft gelatin capsules, aqueous or oily suspensions, emulsions, injectable solutions, dispersible powders or granules, suppositories, lozenges, or syrups, for example.

[0024] As a general guide, the active compounds of the present disclosure are in unit dosage form, or in a form that the patient can self-administer in a single dose. The expression unit dose of the compounds or compositions of the present disclosure can be tablets, capsules, cachets, vials of liquid, powders, granules, lozenges, suppositories, reconstitutable powders, or liquid preparations. Suitable unit doses can be 0.1 to 1000 mg.

[0025] The pharmaceutical compositions of the present disclosure can contain, in addition to the active compounds, one or more adjuvants selected from the following ingredients: fillers (diluents), binders, wetting agents, disintegrants, or excipients, etc. Depending on the method of administration, the compositions can contain 0.1 to 99% by weight of the active compounds.

[0026] In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg to 1000 mg.

[0027] In some embodiments, the pharmaceutical composition contains 0.01 to 99.99% of the aforementioned compound or pharmaceutically acceptable salt thereof or isotopically substituted thereof, based on the total weight of the composition. In some embodiments, the pharmaceutical composition contains 0.1 to 99.9% of the aforementioned compound or pharmaceutically acceptable salt thereof or isotopically substituted thereof. In some embodiments, the pharmaceutical composition contains 0.5 to 99.5% of the aforementioned compound or pharmaceutically acceptable salt thereof or isotopically substituted thereof. In some embodiments, the pharmaceutical composition contains 1 to 99% of the aforementioned compound or pharmaceutically acceptable salt thereof or isotopically substituted thereof. In some embodiments, the pharmaceutical composition contains 2 to 98% of the aforementioned compound or pharmaceutically acceptable salt thereof or isotopically substituted thereof.

[0028] In some embodiments, the pharmaceutical composition contains 0.01% to 99.99% of a pharmaceutically acceptable excipient, based on the total weight of the composition. In some embodiments, the pharmaceutical composition contains 0.1% to 99.9% of a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition contains 0.5% to 99.5% of a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition contains 1% to 99% of a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition contains 2% to 98% of a pharmaceutically acceptable excipient.

[0029] Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients can be inert excipients, granulating agents, disintegrating agents, binding agents, and lubricating agents. The tablets can be uncoated or they can be coated by known techniques to mask the unpleasant taste or odor of the drug or delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.

[0030] Oral preparations can also be provided in the form of soft gelatin capsules wherein the active ingredient is dissolved or suspended in an inert solid diluent or wherein the active ingredient is dissolved or suspended in an oleaginous vehicle.

[0031] Aqueous suspensions contain the active material in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, dispersing agents or wetting agents. The aqueous suspensions can also contain one or more preservatives, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents.

[0032] Oil suspensions can be formulated by suspending the active ingredient in a vegetable oil, or a mineral oil. The oil suspensions can contain a thickening agent. Sweetening agents and flavoring agents can be added to provide a palatable oral preparation. These compositions can be preserved by the addition of an anti-oxidant.

[0033] The pharmaceutical compositions of the present disclosure can also be in the form of oil-in- water emulsions. The oily phase can be a vegetable oil or a mineral oil, or a mixture of these. Suitable emulsifying agents can be naturally-occurring phosphatides, such as soybean lecithin, and mixtures of these. The emulsions can also contain sweetening agents, flavoring agents, preservatives and antioxidants.

[0034] The pharmaceutical compositions of the present disclosure can be in the form of a sterile injectable aqueous solution. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation can also be a sterile injectable oil-in-water microemulsion where the active ingredient is dissolved in the oily phase. The injectable formulations can be injected intramuscularly or subcutaneously, or they can be injected into the blood stream, including intravenously, intravascularly and intracardiacly. Alternatively, solutions and microemulsions are best administered in a manner consistent with the dosage ranges disclosed herein. To maintain the constant concentration of the compounds of the present disclosure in circulation, a continuous intravenous delivery device can be used. An example of such a device is the Deltec CADD-PLUS.TM. Model 5400 intravenous pump.

[0035] The pharmaceutical compositions of the present disclosure can be in the form of a sterile injectable aqueous or oleaginous suspension for intramuscular or subcutaneous administration. This suspension can be formulated according to known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

[0036] The compounds of the present disclosure can be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are

[0037] The compounds of the present disclosure can be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are

[0038] The dosage of the drug depends on a variety of factors, including but not limited to the following: the activity of the particular compound employed, the severity of the disease, the age of the patient, the body weight of the patient, the health status of the patient, the behavior of the patient, the diet of the patient, the time of administration, the mode of administration, the rate of excretion, the combination of drugs, etc. as is well known to those skilled in the art. In addition, the optimal mode of treatment, such as the mode of treatment, the daily dose of the compound, or the type of pharmaceutically acceptable salt, can be verified according to the conventional treatment regimen.

[0039] Explanation of Terms

[0040] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0041] The compounds of the present disclosure can contain all modes of rotation isomers and conformationally restricted states thereof. Also included are atropisomers, the term "atropisomer" is a stereoisomer that results from restricted rotation about a single bond, in which the energy difference due to steric strain or other contributing factors creates a high enough rotational barrier to allow separation of individual conformers. For example, certain compounds of the present disclosure can exist as a mixture of atropisomers (e.g., an equimolar mixture, a mixture enriched in one atropisomer, etc.) or as a purified atropisomer. Non-limiting examples include:

[0042] The compounds and intermediates of the present disclosure can also exist in different tautomeric forms, and all such forms are encompassed within the scope of the present disclosure. The term "tautomers" or "tautomeric forms" refers to different energy structures that can interconvert via a low energy barrier. For example, prototropic tautomers (also known as proton transfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine, lactam-lactim isomerization. An example of a keto-enol equilibrium is shown as follows:

[0043] All tautomeric forms are within the scope of the present disclosure. The naming of the compounds does not exclude any tautomers.

[0044] The compounds of the present disclosure can exist in particular stereoisomeric forms. The term "stereoisomers" refers to isomers having the same structure except they differ in the arrangement of atoms or groups in space. It includes cis- and trans- (or E- and Z-) isomers, (-)- and (+)- isomers, (R)- and (S)- enantiomers, diastereomers, (D)- and (L)- isomers, tautomers, atropisomers, conformational isomers, and mixtures thereof such as racemates, mixtures of diastereomers. Substituents on the compounds of the present disclosure can exist in additional asymmetric atom. All such stereoisomers, as well as mixtures thereof, are encompassed within the scope of the present disclosure. For all carbon-carbon double bonds, both the Z and E isomers are included, even if only one is specifically disclosed. The optically active (-)- and (+)- isomers, (R)- and (S)- enantiomers, and (D)- and (L)- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. One isomer of a certain compound of the present disclosure can be prepared by asymmetric synthesis, or derivatization with a chiral auxiliary, or, when a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group) is present in the molecule, by forming a diastereomeric salt with an appropriate optically active acid or base, and then separating the diastereomeric salt into its components by conventional means, such as fractional crystallization or chromatography. Additionally, the separation of enantiomers and diastereomers is typically accomplished by chromatography.

[0045] In the chemical structure of the compounds of the present disclosure, the bond represents unspecified configuration, i.e. if there is a chiral isomer in the chemical structure, the bond may be or simultaneously contains both configurations; the bond is not specified configuration, i.e. can be Z configuration or E configuration, or simultaneously contains both configurations.

[0046] The compounds of the present disclosure include all suitable isotopic variations of the compounds. The term "isotopic variations" means the replacement of at least one atom with an atom having the same atomic number but an atomic mass different from the predominant atomic isotope usually occurring in nature. Examples of isotopes that can be present in the compounds of the present disclosure include stable and radioactive isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, bromine and iodine, such as 2 H(deuterium, D), 3 H(tritium, T), 2c , 13 C, 14 C, 15 N, 17 O, 18 O, 32 p, 33 p, 33 S, 34 S, 35 S, 36 S, 18 F, 36 Cl, 82 Br, 123 I, 124 I, 125 I, 129 I, and 131 I, in some embodiments deuterium.

[0047] Compared with non-deuterated drugs, deuterated drugs have the advantages of reducing side effects, increasing drug stability, enhancing efficacy, prolonging drug biological half-life, etc. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are included within the scope of the present disclosure. Each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom, wherein the replacement of deuterium can be partial or complete, and the partial replacement of deuterium means that at least one hydrogen is replaced by at least one deuterium.

[0048] "Optionally" or "optional" means that the subsequently described event or circumstance can or can not occur, and the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted with halogen or cyano" means that the C 1-6"Alkyl" means that halogen or cyano can, but need not, be present, and the description includes cases where alkyl is substituted with halogen or cyano and cases where alkyl is not substituted with halogen and cyano.

[0049] "Substituted" means that one or more hydrogen atoms, such as 1 to 6, in some embodiments 1 to 3, hydrogen atoms are independently of each other replaced by a corresponding number of substituents. One of skill in the art can determine, without undue experimentation, through either experiment or theory, what substitutions are possible or impossible. For example, an amino or hydroxyl group with a free hydrogen can not be stable when bound to a carbon atom with an unsaturated (e.g., olefinic) bond.

[0050] "Pharmaceutical composition" means a mixture of one or more of the compounds described herein, or pharmaceutically acceptable salts or prodrugs thereof, with other chemical components, such as pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to a subject, and to facilitate absorption, resulting in the active ingredient exhibiting a biological activity.

[0051] "Pharmaceutically acceptable salt" means a salt of a compound of the disclosure that is safe and effective for use in a mammal, and possesses the desired biological activity. The salts can be prepared from the final isolated and purified compounds, or by reacting a suitable

[0052] The term "pharmaceutically acceptable" as used herein means that these compounds, materials, compositions, and / or dosage forms are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without undue toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio, and effective for their intended use.

[0053] As used herein, the singular forms "a", "an" and "the" include plural references unless the context clearly dictates otherwise.

[0054] When the term "about" is applied to a parameter such as pH, concentration, temperature, etc., it indicates that the parameter can vary by ±10%, and sometimes within ±5%. As will be appreciated by those skilled in the art, numbers are often given only to the nearest integer when the parameter is not critical, and for illustrative purposes only, not as a limitation. DETAILED DESCRIPTION

[0055] The following examples are provided to further illustrate the disclosure, but are not intended to limit the scope of the disclosure.

[0056] EXAMPLES

[0057] The structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR shift (δ) is given in units of 10 -6 (ppm). The determination of NMR is carried out by Bruker AVANCE NEO 500M NMR spectrometer, and the determination solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and the internal standard is tetramethylsilane (TMS).

[0058] The determination of MS is carried out by Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid chromatography-mass spectrometer (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), waters ACQuity UPLC-QD / SQD (manufacturer: waters, MS model: waters ACQuity Qda Detector / waters SQ Detector), THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0059] The high performance liquid chromatography (HPLC) analysis uses Agilent HPLC 1200 DAD, Agilent HPLC 1200 VWD and Waters HPLC e2695-2489 high pressure liquid chromatograph.

[0060] The determination of chiral HPLC analysis uses Agilent 1260 DAD high performance liquid chromatograph.

[0061] The high performance liquid preparation chromatography uses Waters 2767, Waters 2767-SQ Detector 2, Shimadzu LC-20AP and Gilson-281 preparation chromatograph.

[0062] The chiral preparation chromatography uses Shimadzu LC-20AP preparation chromatograph.

[0063] The CombiFlash rapid preparation instrument uses Combiflash Rf200 (TELEDYNE ISCO).

[0064] Thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the specification of silica gel plate used in thin layer chromatography (TLC) is 0.15mm-0.2mm, the specification of product used in thin layer chromatography separation and purification is 0.4mm-0.5mm.

[0065] Silica gel column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as carrier.

[0066] Kinase average inhibition rate and IC 50 The determination of value uses NovoStar enzyme marker (Germany BMG company).

[0067] The known starting materials of the present disclosure can be synthesized or purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals and the like according to the methods known in the art.

[0068] Unless otherwise specified in the examples, the reaction can be carried out under argon atmosphere or nitrogen atmosphere.

[0069] Argon atmosphere or nitrogen atmosphere refers to that the reaction bottle is connected with an argon or nitrogen balloon with a volume of about 1L.

[0070] Hydrogen atmosphere refers to that the reaction bottle is connected with a hydrogen balloon with a volume of about 1L.

[0071] The pressurized hydrogenation reaction uses Parr 3916EKX type hydrogenation instrument and Qinglan QL-500 type hydrogen generator or HC2-SS type hydrogenation instrument.

[0072] The hydrogenation reaction is usually vacuumed, filled with hydrogen, and the operation is repeated for 3 times.

[0073] Microwave reaction uses CEM Discover-S 908860 type microwave reactor.

[0074] Unless otherwise specified in the examples, the solution refers to aqueous solution.

[0075] Unless otherwise specified in the examples, the temperature of the reaction is room temperature.

[0076] The reaction progress in the examples is monitored by thin layer chromatography (TLC), and the developing agent used in the reaction, the eluent system of column chromatography used for purifying compounds and the developing agent system of thin layer chromatography include: A: dichloromethane / methanol system, B: petroleum ether / ethyl acetate, the volume ratio of the solvents is adjusted according to the polarity of the compounds, and a small amount of triethylamine and basic or acidic reagents such as acetic acid can also be added for adjustment.

[0077] Example 1

[0078] 2-amino-4-((S)-5-chloro-3-(((R)-2-(difluoromethyl)tetrahydro-1H-pyrrazin-7a(5H)- yl)methoxy)-7,9-dihydrofuro[3,4-f]quinolin-6-yl)-7-fluorobenzo[b]thiophene-3- carbonitrile 1

[0079] First Step

[0080] 2-(difluoromethyl)-5-oxotetrahydro-1H-pyrrazine-7a(5H)-carboxylic acid ethyl ester 1b

[0081] Ethyl 2,5-dioxotetrahydro-1H-pyrrazine-7a(5H)-carboxylate 1a (18.08 g, 85.60 mmol, Aikchem) and (triphenylphosphoranylidene)difluoroacetic acid inner salt (67.1 g, 188.3, mmol, Aikchem) were dissolved in N,N-dimethylformamide (350 mL) and stirred at 85 °C under nitrogen atmosphere for 17 hours. After the reaction solution was cooled to room temperature, saturated sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate (100 mL x 3). The combined organic phase was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using eluent system B to obtain the title compound 1b (4.83 g, yield: 23%).

[0082] MS m / z (ESI): 246.1 [M+1].

[0083] Second Step

[0084] (2-(difluoromethyl)tetrahydro-1H-pyrrazine-7a(5H)-yl)methanol 1c

[0085] (R)-(2-(difluoromethyl)tetrahydro-1H-pyrrazine-7a(5H)-yl)methanol 1c-1

[0086] (S)-(2-(difluoromethyl)tetrahydro-1H-pyrrazine-7a(5H)-yl)methanol 1c-2

[0087] Compound 1b (4.83 g, 19.7 mmol) was dissolved in tetrahydrofuran (250 mL), and 1M diisobutylaluminum hydride in tetrahydrofuran (98.5 mL) was added under ice bath. After the temperature was naturally recovered and stirred for 5 hours, sodium sulfate decahydrate was added and stirred for 0.5 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain the title crude compound 1c (2.1 g).

[0088] MS m / z (ESI): 190.1 [M+1].

[0089] Isomer mixture 1c (1 g) was separated by chiral column (Shimadzu LC-20AP, column: DAICEL CHIRALPAK IG, 20*250mm, 5um; mobile phase A: n-hexane, mobile phase B: ethanol (0.1% 7M NH3 in MeOH), gradient ratio: A:B: 80:20, flow rate: 20 mL / min) to give title compound 1c-1 (450 mg, yield: 45%) and 1c-2 (450 mg, yield: 45%).

[0090] 1c-1 Chiral HPLC analysis: retention time 3.220 min, purity: 99% (column: Agilent 1260 DAD, column: CHIRALPAK IG 150*4.6mm, 5um; mobile phase A: ethanol (0.1% diethylamine, mobile phase B: n-hexane), gradient ratio: A:B: 20:80, flow rate: 1 mL / min).

[0091] 1c-2 Chiral HPLC analysis: retention time 4.330 min, purity: 99% (column: Agilent 1260 DAD, column: CHIRALPAK IG 150*4.6mm, 5um; mobile phase A: ethanol (0.1% diethylamine, mobile phase B: n-hexane), gradient ratio: A:B: 20:80, flow rate: 1 mL / min).

[0092] Third step

[0093] 6-bromo-5-chloro-3-(ethylsulfonyl)-7,9-dihydrofuro[3,4-f]quinazoline 1e

[0094] 6-bromo-3-(ethylthio)-5-chloro-7,9-dihydrofuro[3,4-f]quinazoline 1d (1.94 g, 5.61 mmol, prepared by the method disclosed in the preparation 300 on page 192 of the specification of patent application “WO2023183585A1”) was dissolved in dichloromethane (50 mL), m-chloroperoxybenzoic acid (3.42 g, 16.8 mmol, 85% purity) was added, the reaction was stirred for 2 hours, saturated sodium bicarbonate solution was added to the reaction solution, extracted with dichloromethane (30 mL x 2), the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was slurried with ethyl acetate, filtered, and the filter cake was dried to give the crude title compound 1e (1.76 g), which was used directly in the next step without purification.

[0095] MS m / z (ESI): 376.9 [M+1].

[0096] Fourth step

[0097] (R)-6-bromo-5-chloro-3-((2-(difluoromethyl)tetrahydro-1H-pyrrazin-7a(5H)-yl)methoxy)- 7,9-dihydrofuro[3,4-f]quinoline 1f

[0098] Compound 1c-1 (142 mg, 750 μmol) was dissolved in tetrahydrofuran (10 mL), 2M bis(trimethylsilyl)aminium sodium tetrahydrofuran solution (374 μL) was added under ice bath, after 30 minutes of stirring at the temperature, compound 1e (225 mg, 596 μmol) tetrahydrofuran solution (5 mL) was added under ice bath, the reaction was carried out at 50°C for 1 hour, after the reaction solution was cooled to room temperature, saturated ammonium chloride solution was added to quench, extracted with ethyl acetate (20 mL x 2), the combined organic phase was dried with anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1f (176 mg, yield: 62.4%).

[0099] MS m / z (ESI): 472.2 [M+1].

[0100] Fifth step

[0101] (4-(5-chloro-3-(((R)-2-(difluoromethyl)tetrahydro-1H-pyrrazin-7a(5H)-yl)methoxy)-7,9- dihydrofuro[3,4-f]quinolin-6-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamic acid tert-butyl ester 1g

[0102] Compound 1f (176 mg, 372 μmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-7- fluoro benzo [b] thiophen-2-yl) carbamic acid tert-butyl ester (300 mg, 742 μmol) were dissolved in 1.4-dioxane (10 mL), cesium carbonate (303 mg, 0.93 mmol) and 1,1'- bis(diphenylphosphino)ferrocene palladium (II) dichloride (55 mg, 75 μmol) were added, the reaction was carried out at 100°C for 2 hours under nitrogen atmosphere, the reaction solution was concentrated under reduced pressure, the residue was dissolved in dichloromethane, water was added, the aqueous phase was extracted with dichloromethane (15 mL x 2), the combined organic phase was dried with anhydrous sodium sulfate, after removing the drying agent by filtration, the filtrate was concentrated under reduced pressure, the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1g (211 mg, yield: 82.8%).

[0103] MS m / z (ESI): 684.5 [M+1].

[0104] Sixth step

[0105] 2-amino-4-(5-chloro-3-(((R)-2-(difluoromethyl)tetrahydro-1H-pyrrozin-7a(5H)-yl)methoxy)- 7,9-dihydrofuro[3,4-f]quinazolin-6-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile 1h

[0106] 2-amino-4-(5-chloro-3-(((R)-2-(difluoromethyl)tetrahydro-1H-pyrrozin-7a(5H)-yl)methoxy)- 7,9-dihydrofuro[3,4-f]quinazolin-6-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile 1h

[0107] Compound 1g (211 mg, 308 μmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added, the reaction was stirred for 1 hour, the reaction solution was concentrated under reduced pressure, the residue was dissolved in dichloromethane, the pH was adjusted to >7 with saturated sodium bicarbonate, the aqueous phase was extracted with dichloromethane (10 mL x 2), the combined organic phase was dried with anhydrous sodium sulfate, after removing the drying agent by filtration, it was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (Waters-2545, column: YMC Triart-Exrs C18, 30 x 150 mm, 5 μm; mobile phase: aqueous phase (10 mmol / L ammonium bicarbonate) and acetonitrile, gradient ratio: acetonitrile 32%-45%, flow rate: 30 mL / min) to obtain the title compound 1h (95 mg, yield: 52.7%).

[0108] Isomer mixture 1h (90 mg) was separated by chiral column (Gilson-281, column: CHIRALPAK IG, 20 x 250 mm, 5 μm; mobile phase A: n-hexane, mobile phase B: ethanol (0.1% 7M ammonium methanol solution), gradient ratio: A:B:70:30, flow rate: 20 mL / min), and the compound with shorter retention time, i.e. 1 (30 mg, yield: 33.3%) was collected.

[0109] MS m / z (ESI): 584.2 [M+1].

[0110] 1H NMR (500 MHz, DMSO-d6): δ 9.49 (s, 1H), 8.10-8.08 (m, 2H), 7.26-7.24 (m, 1H), 7.17-7.13 (m, 1H), 5.56-5.55 (m, 2H), 4.90-4.87 (m, 1H), 4.74-4.71 (m, 1H), 4.38-4.36 (m, 1H), 4.24-4.22 (m, 1H), 3.71-3.68 (m, 1H), 3.32-3.31 (m, 1H), 3.04-3.00 (m, 1H), 2.72-2.69 (m, 1H), 2.59-2.55 (m, 1H), 2.48-2.45 (m, 1H), 2.06-2.00 (m, 1H), 1.92-1.78 (m, 3H).

[0111] Example 2

[0112] 2-amino-4-((S)-5-chloro-3-(((S)-2-(difluoromethylidene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,9-dihydrofuro[3,4-f]quinolin-6-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile 2

[0113] The title compound 2 (5 mg, yield: 8.4%) was obtained by using the synthetic route four to six in Example 1, replacing the fourth step raw material compound 1c-1 with compound 1c-2.

[0114] MS m / z (ESI): 584.2 [M+1].

[0115] 1 H NMR (500 MHz, DMSO-d6): δ 9.49 (s, 1H), 8.10-8.08 (m, 2H), 7.26-7.24 (m, 1H), 7.17-7.13 (m, 1H), 5.56-5.55 (m, 2H), 4.90-4.87 (m, 1H), 4.74-4.71 (m, 1H), 4.38-4.36 (m, 1H), 4.24-4.22 (m, 1H), 3.71-3.68 (m, 1H), 3.32-3.31 (m, 1H), 3.04-3.00 (m, 1H), 2.72-2.69 (m, 1H), 2.59-2.55 (m, 1H), 2.48-2.45 (m, 1H), 2.06-2.00 (m, 1H), 1.92-1.78 (m, 3H).

[0116] Example 3 (control)

[0117] The synthetic route four to six in Example 1 was adopted, and the raw material compound 1c-1 in the fourth step was replaced by ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a-yl)methanol to obtain the title compound 3 (300 mg, yield: 22.2%).

[0118] MS m / z (ESI): 554.5 [M+1].

[0119] 1 H NMR (500 MHz, CD3OD): δ 9.37 (s, 1H), 7.22 (dd, 1H), 7.06 (t, 1H), 5.62 (q, 2H), 5.44-5.20 (m, 1H), 4.98 (dt, 1H), 4.85-4.77 (m, 1H), 4.49 (d, 1H), 4.40 (d, 1H), 3.28-3.15 (m, 2H), 3.09-3.01 (m, 1H), 2.56-2.35 (m, 1H), 2.35-2.16 (m, 3H), 2.14-1.88 (m, 3H).

[0120] Biological evaluation

[0121] Test Example 1: 3D cell proliferation inhibition experiment

[0122] This experiment used the cells listed in the table below to evaluate the inhibitory effect of the compounds on the proliferation of KRAS mutant or KRAS amplified cells.

[0123] On the first day of the experiment, cells growing well, reaching 70%-80% confluence, were digested, resuspended with culture medium, and the cell density was adjusted to the required. 90 μL of cell suspension was added to each well of a U-shaped low-adsorption 96-well plate (Corning, 7007), and the cell density was as shown in the table above. After centrifugation of the cell plate at 2500 rpm for 5 minutes, it was placed in an incubator for 24 hours. On the second day, 20 mM of the test compound dissolved in DMSO was diluted with DMSO to the first concentration of 2 or 0.2 mM, and then diluted by 5 times gradient, a total of 9 concentration points, and the control well was DMSO. Then the gradient-diluted compound was further diluted 20 times with culture medium. 10 μL of the test compound diluted with culture medium was added to each well of the cell plate. The final concentration of the compound was 9 concentration points starting from 10 or 1 μM with 5 times gradient dilution. The cell well containing 0.5% DMSO was set as the solvent control well, and the well containing only culture medium and 0.5% DMSO was the blank control well. After centrifugation of the cell plate at 2000 rpm for 3 minutes, it was placed in an incubator for 5 days. On the seventh day, the 96-well cell culture plate was taken out, 50 μL of luminescent cell viability detection reagent 3D Cell Viability Assay (Promega, G9683), after shaking for 25 minutes at room temperature, blow and suck to mix and take out 100 μL per hole to transfer to white non-bottom OptiPlate TM -96-well plate (PerkinElmer, 6005290), using a multifunctional microplate luminometer (PerkinElmer, EnVision2105) to read the luminescence signal value.

[0124] The inhibition rate was calculated using the following formula: inhibition rate = (luminescence value 溶媒对照孔 - luminescence value 受试化合物 ) / (luminescence value 溶 媒对照孔 - luminescence value 空白对照孔 ) x 100%. The IC 50 values of the inhibitory activity of the compounds were calculated using Graphpad Prism software, and the results are shown in the table below.

[0125] Table 1, 3D cell proliferation inhibition activity data (unit: nM)

[0126] Conclusion: The compounds of the present disclosure have good inhibitory effect on the 3D proliferation of the above-mentioned cells.

[0127] Test Example 2, Pharmacokinetic Evaluation

[0128] 1. Abstract

[0129] SD rats were used as test animals, and LC-MS / MS method was used to determine the drug concentration in rat plasma at different time points after i.g. administration of the compound of the example, to study the pharmacokinetic behavior of the compound of the present disclosure in rats, and to evaluate its pharmacokinetic characteristics.

[0130] 2. Test Plan

[0131] 2.1. Test Drug

[0132] Compounds 1, 2 and 3.

[0133] 2.2. Test Animals

[0134] 8 female rats were provided by Vantianlihua Experimental Animal Technology Co., Ltd., with production license SCXK (Zhejiang) 2024-0001.

[0135] 2.3. Drug Preparation

[0136] Compound 1: A certain amount of test compound was weighed, and 5% DMSO + 10% HS15 + 17% (w / v) SBE-β-CD was added to prepare 2.0 mg / mL and 6.0 mg / mL colorless and clear solutions.

[0137] Compound 2 and 3: A certain amount of test compound was weighed, 5% DMSO + 10% HS15 + 17% (w / v) SBE-β-CD was added, and a 2.0 mg / mL yellow suspension was prepared.

[0138] 2.4, Administration

[0139] Compound 1: The administration dose was 20.0 mg / kg and 60 mg / kg, and the administration volume was 10 mL / kg.

[0140] Compound 2 and 3: The administration dose was 20.0 mg / kg, and the administration volume was 10 mL / kg.

[0141] 3, Operation

[0142] Before administration and at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, and 24.0 hours after administration, 0.2 mL of blood was taken from the eye socket, placed in an EDTA-K2 anticoagulant tube, centrifuged at 10,000 rpm for 2 minutes (4°C), and the plasma was separated within 1 hour and stored at -20°C or -80°C for testing. The blood collection to centrifugation process was operated under ice bath conditions.

[0143] The content of the test compound in the rat plasma after administration of different compounds was determined: Compound 1 (20 mg / kg / 60 mg / kg), 25 μL / 20 μL of rat plasma sample at each time point after administration was taken, 200 μL / 250 μL of internal standard acetonitrile solution was added, vortex mixed, and centrifuged at 4000 rpm for 15 minutes. The supernatant was analyzed by LC-MS / MS.

[0144] Compound 2, 20 μL of rat plasma sample at each time point after administration was taken, 200 μL of internal standard acetonitrile solution was added, vortex mixed, and centrifuged at 4000 rpm for 15 minutes. The supernatant was analyzed by LC-MS / MS.

[0145] Compound 3: 20 μL of rat plasma sample at each time point after administration was taken, 250 μL of internal standard acetonitrile solution was added, vortex mixed, and centrifuged at 4000 rpm for 15 minutes. The supernatant was analyzed by LC-MS / MS.

[0146] 4, Pharmacokinetic parameter results

[0147] Table 2, Pharmacokinetic parameters of the compounds of the present disclosure in rats

[0148] Conclusion: The compounds of the present disclosure have high blood drug concentrations and high exposure in rats, and have pharmacokinetic advantages.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, which is selected from the following compounds:

2. A compound or a salt thereof selected from:

3. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

4. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 3, in the preparation of a medicament for inhibiting KRAS amplification and / or KRAS mutant activity; the KRAS mutant is preferably KRAS G12D and / or KRAS G12V mutation.

5. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 3 in the preparation of a medicament for treating and / or preventing cancer, wherein the cancer is preferably selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial carcinoma, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma and glioblastoma; more preferably selected from pancreatic cancer, colorectal cancer and non-small cell lung cancer.

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