PRMT5-MTA inhibitor

Abstract

The present invention provides a compound as a PRMT5-MTA inhibitor, which is a compound represented by formula (I) or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof. The present invention also provides a pharmaceutical composition comprising the compound and its use in the treatment of cancer.

Description

[0001] This invention claims the priority of Chinese Patent Application No. 202210757379.5, filed on June 29, 2022. Technical Field

[0002] This invention belongs to the field of medicine and particularly relates to PRMT5-MTA inhibitors. Background Art

[0003] Protein arginine methyltransferase (PRMT) can methylate histones and non-histones to participate in the regulation of biological processes such as gene transcription, signal transduction, protein stability, cell proliferation, differentiation, apoptosis, and tumor formation. Currently, 11 PRMT family members have been discovered. According to the different ways of catalyzing arginine methylation, they can be divided into types I, II, and III. Among them, PRMT5 belongs to type II, and its catalytic form is symmetric dimethylation.

[0004] As an epigenetic enzyme, PRMT5 participates in various biological processes, including transcriptional regulation, RNA metabolism, ribosome biosynthesis, and cell cycle regulation. The PRMT5 protein is overexpressed in various cancer types, including B and T cell lymphomas, metastatic melanoma, neuroblastoma, glioblastoma, ovarian cancer, breast cancer, etc. More and more evidence indicates that it plays an important role in tumorigenesis and development. On this basis, PRMT5 inhibitors have become a research hotspot for tumor treatment drugs.

[0005] Early PRMT5 inhibitors can be divided into two categories. One is substrate-competitive inhibitors, with the representative drug being GSK3326595; the other is SAM-competitive inhibitors, with the representative drug being JNJ64619178. Both of these two types of drugs have strong inhibitory activity against PRMT5 and show strong anti-tumor activity. However, due to their strong inhibitory activity against PRMT5 in both normal cells and tumor cells, strong hematotoxicity has been observed, which limits their clinical application and thus affects their clinical treatment effects.

[0006] In 2016, a paper published in Science revealed the synthetic lethality between MTAP deficiency and PRMT5. MTAP has a high deletion rate in a variety of solid tumors, including pancreatic cancer, glioma, etc. MTAP is an intracellular MTA degrading enzyme. Since MTAP deficiency can lead to the intracellular accumulation of MTA, and MTA can compete with the functional substrate methyl donor SAM of PRMT5 for binding to PRMT5, thereby inhibiting the function of PRMT5. Due to the specific accumulation of MTA in MTAP-deficient tumor cells, by strengthening the binding inhibition of MTA and PRMT5, it is possible to specifically inhibit the activity of PRMT5 in tumor cells, while having a weak inhibitory effect on the PRMT5 activity of normal cells, thus providing a therapeutic safety window and reducing toxicity while ensuring anti-tumor efficacy. Currently, MTA cooperative PRMT5 inhibitors have obtained preclinical validation data. The research and development of MTA cooperative PRMT5 inhibitors have great potential in the treatment of MTAP-deficient tumors.

[0007] Currently, despite the progress in PRMT5 research, Mirati Therapeutics reported its PRMT5-MTA inhibitor MRTX1719, but there is still a lack of effective and selective PRMT5-MTA inhibitors. Summary of the Invention

[0008] In one aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

[0009]

[0010] Wherein,

[0011] R1 is selected from C 1-6 alkyl, C 1-6 haloalkyl, -(CH2) 1-4 -NH2, -(CH2) 1-4 -OH, C 3-7 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl, which may optionally be substituted by 1, 2, 3, 4 or 5 R*; R* is selected from halogen, CN, OH, NH2, C 1-6 alkyl, C 3-6 cycloalkyl or C 1-6 haloalkyl;

[0012] R2 is selected from CN, OH or NH2;

[0013] R3 is selected from H, -O-C 3-7 cycloalkyl or -O-3- to 7-membered heterocyclic group;

[0014] R4 is selected from H or -(CH2) 1-4 -3- to 7-membered heterocyclic group, wherein said -(CH2) 1-4 -3- to 7-membered heterocyclic group may optionally be substituted by 1, 2, 3, 4 or 5 substituents selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl;

[0015] R5 is selected from H, -C≡C-R 5a or -P(O)-(C 1-6 alkyl)2;

[0016] R 5a is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl, which may optionally be substituted by 1, 2, 3, 4 or 5 R#; R# is selected from halogen, CN, OH, NH2, C 1-6 alkyl or C 1-6 haloalkyl;

[0017] R6 is halogen;

[0018] R7 is selected from H, halogen or C 1-6 alkyl;

[0019] R8 is selected from H, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy or C 1-6 haloalkoxy.

[0020] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention and optionally a pharmaceutically acceptable excipient.

[0021] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient, which further comprises other therapeutic agents.

[0022] In another aspect, the present invention provides the use of a compound of the present invention in the preparation of a drug for treating and / or preventing diseases mediated by PRMT5 arginine methyltransferase.

[0023] In another aspect, the present invention provides a method for treating and / or preventing diseases mediated by PRMT5 arginine methyltransferase in a subject, comprising administering to the subject a compound of the present invention or a composition of the present invention.

[0024] In another aspect, the present invention provides a compound or a composition of the present invention for treating and / or preventing diseases mediated by PRMT5 arginine methyltransferase.

[0025] In a specific embodiment, the diseases treated by the present invention include cancers selected from the following: acoustic neuroma, adenocarcinoma, adrenal cancer, anal cancer, angiosarcoma (such as lymphangiosarcoma, lymphangioendotheliosarcoma, hemangioma), appendiceal cancer, benign monoclonal gammopathy, cholangiocarcinoma, bladder cancer, brain cancer (such as meningioma, glioma, such as astrocytoma, oligodendroglioma, medulloblastoma), bronchial cancer, carcinoid tumor, cervical cancer (such as cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (such as colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial cancer, ependymoma, endothelial sarcoma (such as Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial cancer (such as uterine cancer, uterine sarcoma), esophageal cancer (such as esophageal adenocarcinoma, Barrett's adenocarcinoma), Ewing's sarcoma, eye cancer (such as intraocular melanoma, retinoblastoma), eosinophilia, gallbladder cancer, gastric cancer (such as gastric adenocarcinoma), gastrointestinal stromal tumor (GIST), head and neck cancer (such as head and neck squamous cell carcinoma, oral cancer (such as oral squamous cell carcinoma, laryngeal cancer (such as laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer))), hematopoietic system cancers (such as leukemia, such as acute lymphoblastic leukemia (ALL) (such as B-cell ALL, T-cell ALL), acute myeloid leukemia (AML) (such as B-cell AML, T-cell AML), chronic myeloid leukemia (CML) (such as B-cell CML, T-cell CML), chronic lymphocytic leukemia (CLL) (such as B-cell CLL, T-cell CLL), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), marginal zone B-cell lymphoma (such as mucosa-associated lymphoid tissue (MALT) lymphoma, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma, and primary central nervous system (CNS) lymphoma; and T-cell non-Hodgkin lymphoma, such as precursor T lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (such as cutaneous T-cell lymphoma (such as mycosis fungoides, Sézary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); a mixture of one or more of the above leukemias / lymphomas;Multiple myeloma (MM), heavy chain diseases (such as alpha chain disease, gamma chain disease, mu chain disease), hemangioblastoma, inflammatory myofibroblastic tumor, immunocytic amyloidosis, renal cancer (such as nephroblastoma, renal cell carcinoma), liver cancer (such as hepatocellular carcinoma, malignant hepatocellular carcinoma), lung cancer (such as bronchial carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma, leiomyosarcoma (LMS), mastocytosis (such as systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative diseases (MPD) (such as polycythemia vera (PV), essential thrombocythemia (ET), idiopathic myelofibrosis with extramedullary hematopoiesis (AMM), chronic idiopathic myelofibrosis, chronic myelogenous leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES), neuroblastoma, neurofibroma (such as neurofibromatosis type 1 or 2, schwannomatosis), neuroendocrine carcinoma (such as gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer (such as cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma, penile cancer.;

[0026] Other objects and advantages of the present invention will be apparent to those skilled in the art from the following specific embodiments, examples and claims.

[0027] Definitions

[0028] Chemical Definitions

[0029] The definitions of specific functional groups and chemical terms are described in more detail below.

[0030] When a numerical range is listed, each value and sub-range within the range are intended to be included. For example, "C 1-6 alkyl" includes C1, C2, C3, C4, C5, C6, C 1-6 、C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-5 、C 2-4 、C 2-3 、C 3-6 、C 3-5 、C 3-4 、C 4-6 、C 4-5 and C 5-6 alkyl.

[0031] "C 1-6 alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms. In some embodiments, C 1-4 alkyl and C1-2 An alkyl group is preferred. C 1-6 Examples of alkyl groups include: methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). The term "C 1-6 alkyl" also includes heteroalkyls in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkyl group may be optionally substituted with one or more substituents, e.g., substituted with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. Conventional alkyl abbreviations include: Me (-CH3), Et (-CH2CH3), iPr (-CH(CH3)2), nPr (-CH2CH2CH3), n-Bu (-CH2CH2CH2CH3), or i-Bu (-CH2CH(CH3)2).

[0032] "C 1-6 alkylene" refers to a divalent group formed by removing another hydrogen of the C 1-6 alkyl, and may be substituted or unsubstituted. In some embodiments, C 1-4 alkylene, C 2-4 alkylene, and C 1-3 alkylene are preferred. Unsubstituted alkylene includes, but is not limited to: methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-), pentylene (-CH2CH2CH2CH2CH2-), hexylene (-CH2CH2CH2CH2CH2CH2-), and the like. Exemplary substituted alkylene, e.g., alkylene substituted with one or more alkyl groups (methyl), includes, but is not limited to: substituted methylene (-CH(CH3)-, -C(CH3)2-), substituted ethylene (-CH(CH3)CH2-, -CH2CH(CH3)-, -C(CH3)2CH2-, -CH2C(CH3) 2- ), substituted propylene (-CH(CH3)CH2CH2-, -CH2CH(CH3)CH2-, -CH2CH2CH(CH3)-, -C(CH3)2CH2CH2-, -CH2C(CH3)2CH2-, -CH2CH2C(CH3)2-), and the like.

[0033] "Halogenated" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0034] Thus, "C 1-6 haloalkyl" refers to the above-mentioned "C 1-6 alkyl" which is substituted by one or more halogen groups. In some embodiments, C 1-4 haloalkyl is particularly preferred, and more preferably C 1-2 haloalkyl. Exemplary haloalkyls include, but are not limited to: -CF3, -CH2F, -CHF2, -CHFCH2F, -CH2CHF2, -CF2CF3, -CCl3, -CH2Cl, -CHCl2, 2,2,2-trifluoro-1,1-dimethylethyl, and the like. The haloalkyl group can be substituted at any available attachment point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0035] "C 1-6 alkoxy" refers to an -OR group, where R is the C 1-6 alkyl defined above. C 1-4 alkoxy is preferred.

[0036] "C 1-6 haloalkoxy" refers to the above-mentioned "C 1-6 alkoxy" which is substituted by one or more halogen groups. In some embodiments, C 1-4 haloalkoxy is particularly preferred, and more preferably C 1-2 haloalkoxy. Exemplary haloalkyls include, but are not limited to: -OCF3, -OCH2F, -OCHF2, -OCHFCH2F, -OCH2CHF2, -OCF2CF3, -OCCl3, -OCH2Cl, -OCHCl2, and the like. The haloalkyl group can be substituted at any available attachment point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0037] "C 3-10 cycloalkyl" refers to a non-aromatic cycloaliphatic hydrocarbon group having 3 to 10 ring carbon atoms and zero heteroatoms. In some embodiments, C 4-10 cycloalkyl, C 3-7 cycloalkyl, C 3-6 cycloalkyl and C 3-5 cycloalkyl are particularly preferred, and more preferably C 5-6Cycloalkyl. The cycloalkyl also includes a ring system in which the above cycloalkyl ring is fused with one or more aryl or heteroaryl groups, where the point of attachment is on the cycloalkyl ring, and in such cases, the number of carbons continues to represent the number of carbons in the cycloalkyl system. Exemplary cycloalkyls include, but are not limited to: cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), and so on. The cycloalkyl group may be optionally substituted with one or more substituents, for example, substituted with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0038] "3- to 10-membered heterocyclic group" means a group of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. In a heterocyclic group containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom as long as the valence allows. In some embodiments, a 4- to 9-membered heterocyclic group is preferred, which is a 4- to 9-membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 5- to 8-membered heterocyclic group is preferred, which is a 5- to 8-membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 3- to 8-membered heterocyclic group is preferred, which is a 3- to 8-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms; a 3- to 7-membered heterocyclic group is preferred, which is a 3- to 7-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; a 4- to 7-membered heterocyclic group is preferred, which is a 4- to 7-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; a 4- to 6-membered heterocyclic group is preferred, which is a 4- to 6-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; more preferably, a 5- to 6-membered heterocyclic group, which is a 5- to 6-membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms. The heterocyclic group also includes a ring system in which the above heterocyclic group ring is fused to one or more cycloalkyl groups, wherein the point of attachment is on the cycloalkyl ring, or a ring system in which the above heterocyclic group ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclic group ring; and in such cases, the number of ring members continues to represent the number of ring members in the heterocyclic group ring system. Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to: aziridinyl, oxiranyl, thiorenyl. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to: azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to: tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrol-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to: dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to: triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to: piperidinyl, tetrahydropyranyl, dihydropyridyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to: piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to: triazinanyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to: azepanyl, oxepanyl, and thiepanyl.Exemplary 5-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 5,6-bicyclic heterocyclic groups) include, but are not limited to: indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 6,6-bicyclic heterocyclic groups) include, but are not limited to: tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like. The heterocyclic groups may be optionally substituted with one or more substituents, for example, substituted with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0039] "C 6-10 aryl" refers to a group having a monocyclic or polycyclic (e.g., bicyclic) 4n+2 aromatic ring system with 6-10 ring carbon atoms and zero heteroatoms (e.g., having 6 or 10 π electrons shared in a cyclic arrangement). In some embodiments, the aryl has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, the aryl has ten ring carbon atoms ("C 10 aryl"; e.g., naphthyl, e.g., 1-naphthyl and 2-naphthyl). The aryl also includes ring systems in which the above aryl ring is fused to one or more cycloalkyl or heterocyclic groups, and the point of attachment is on the aryl ring, in which case the number of carbon atoms continues to represent the number of carbon atoms in the aryl ring system. The aryl groups may be optionally substituted with one or more substituents, for example, substituted with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0040] "5-10 membered heteroaryl" refers to a group having a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system with ring carbon atoms and 1-4 ring heteroatoms (e.g., having 6 or 10 π electrons shared in a cyclic arrangement), wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In a heteroaryl containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom as long as the valence allows. The heteroaryl bicyclic system can include one or more heteroatoms in one or both rings. The heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl or heterocyclic groups, and the point of attachment is on the heteroaryl ring. In this case, the number of carbon atoms continues to represent the number of carbon atoms in the heteroaryl ring system. In some embodiments, 5-9 membered heteroaryl is preferred, which is a 5-9 membered monocyclic or bicyclic 4n+2 aromatic ring system with ring carbon atoms and 1-4 ring heteroatoms. In other embodiments, 5-6 membered heteroaryl is particularly preferred, which is a 5-6 membered monocyclic or bicyclic 4n+2 aromatic ring system with ring carbon atoms and 1-4 ring heteroatoms. Exemplary 5-membered heteroaryls containing one heteroatom include, but are not limited to: pyrrolyl, furyl, and thienyl. Exemplary 5-membered heteroaryls containing two heteroatoms include, but are not limited to: imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryls containing three heteroatoms include, but are not limited to: triazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl), and thiadiazolyl. Exemplary 5-membered heteroaryls containing four heteroatoms include, but are not limited to: tetrazolyl. Exemplary 6-membered heteroaryls containing one heteroatom include, but are not limited to: pyridyl. Exemplary 6-membered heteroaryls containing two heteroatoms include, but are not limited to: pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryls containing three or four heteroatoms include, but are not limited to: triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryls containing one heteroatom include, but are not limited to: azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryls include, but are not limited to: indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuryl, isobenzofuryl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryls include, but are not limited to: naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. The heteroaryl group can be optionally substituted by one or more substituents, e.g., by 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0041] Alkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, etc. as defined herein are optionally substituted groups.

[0042] Exemplary substituents on a carbon atom include, but are not limited to: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR aa , -ON(R bb )2, -N(R bb )2, -N(R bb )3 + X - , -N(OR cc )R bb , -SH, -SR aa , -SSR cc , -C(=O)R aa , -CO2H, -CHO, -C(OR cc )2, -CO2R aa , -OC(=O)R aa , -OCO2R aa , -C(=O)N(R bb )2, -OC(=O)N(R bb )2, -NR bb C(=O)R aa , -NR bb CO2R aa , -NR bb C(=O)N(R bb )2, -C(=NR bb )R aa , -C(=NR bb )OR aa , -OC(=NR bb )R aa , -OC(=NR bb )OR aa , -C(=NR bb )N(R bb )2, -OC(=NR bb )N(R bb )2, -NR bb C(=NR bb )N(R bb )2, -C(=O)NR bb SO2R aa , -NR bb SO2R aa , -SO2N(R bb )2, -SO2R aa , -SO2OR aa , -OSO2R aa , -S(=O)R aa , -OS(=O)R aa , -Si(R aa)3, -OSi(R aa )3, -C(=S)N(R bb )2, -C(=O)SR aa , -C(=S)SR aa , -SC(=S)SR aa , -SC(=O)SR aa , -OC(=O)SR aa , -SC(=O)OR aa , -SC(=O)R aa , -P(=O)2R aa , -OP(=O)2R aa , -P(=O)(R aa )2, -OP(=O)(R aa )2, -OP(=O)(OR cc )2, -P(=O)2N(R bb )2, -OP(=O)2N(R bb )2, -P(=O)(NR bb )2, -OP(=O)(NR bb )2, -NR bb P(=O)(OR cc )2, -NR bb P(=O)(NR bb )2, -P(R cc )2, -P(R cc )3, -OP(R cc )2, -OP(R cc )3, -B(R aa )2, -B(OR cc )2, -BR aa (OR cc ), alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd groups;

[0043] Or two geminal hydrogens on a carbon atom are substituted by the groups =O, =S, =NN(R bb )2, =NNR bb C(=O)R aa , =NNR bb C(=O)OR aa , =NNR bb S(=O)2R aa , =NR bb or =NOR cc substituted;

[0044] R aa Each of which is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl, or two R aa groups combine to form a heterocyclic group or a heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd groups;

[0045] R bb Each of which is independently selected from: hydrogen, -OH, -OR aa , -N(R cc )2, -CN, -C(=O)R aa , -C(=O)N(R cc )2, -CO2R aa , -SO2R aa , -C(=NR cc )OR aa , -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc , -SOR aa , -C(=S)N(R cc )2, -C(=O)SR cc , -C(=S)SR cc , -P(=O)2R aa , -P(=O)(R aa )2, -P(=O)2N(R cc )2, -P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl, or two R bb groups combine to form a heterocyclic group or a heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd groups;

[0046] R cc Each of which is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl, or two R cc groups combine to form a heterocyclic group or a heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd groups;

[0047] R ddEach is independently selected from: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee 、-ON(R ff )2、-N(R ff )2,、-N(R ff )3 + X - 、-N(OR ee )R ff 、-SH、-SR ee 、-SSR ee 、-C(=O)R ee 、-CO2H、-CO2R ee 、-OC(=O)R ee 、-OCO2R ee 、-C(=O)N(R ff )2、-OC(=O)N(R ff )2、-NR ff C(=O)R ee 、-NR ff CO2R ee 、-NR ff C(=O)N(R ff )2、-C(=NR ff )OR ee 、-OC(=NR ff )R ee 、-OC(=NR ff )OR ee 、-C(=NR ff )N(R ff )2、-OC(=NR ff )N(R ff )2、-NR ff C(=NR ff )N(R ff )2、-NR ff SO2R ee 、-SO2N(R ff )2、-SO2R ee 、-SO2OR ee 、-OSO2R ee 、-S(=O)R ee 、-Si(R ee )3、-OSi(R ee )3、-C(=S)N(R ff )2、-C(=O)SR ee 、-C(=S)SR ee 、-SC(=S)SR ee 、-P(=O)2Ree 、 -P(=O)(R ee )2, -OP(=O)(R ee )2, -OP(=O)(OR ee )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg groups, or two geminal R dd substituents may combine to form =O or =S;

[0048] Each R ee is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic group and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg groups;

[0049] Each R ff is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl, or two R ff groups combine to form a heterocyclic group or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg groups;

[0050] Each R gg is independently: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC 1-6 alkyl, -ON(C 1-6 alkyl)2, -N(C 1-6 alkyl)2, -N(C 1-6 alkyl)3 + X - , -NH(C 1-6 alkyl)2 + X - , -NH2(C 1-6 alkyl) + X - , -NH3 + X - , -N(OC 1-6 alkyl)(C 1-6 alkyl), -N(OH)(C 1-6 alkyl), -NH(OH), -SH, -SC 1-6 alkyl, -SS(C 1-6 alkyl), -C(=O)(C 1-6alkyl), -CO2H, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 alkyl)2, -OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 alkyl)2, -NHC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -C(=NH)O(C 1-6 alkyl), -OC(=NH)(C 1-6 alkyl), -OC(=NH)OC 1-6 alkyl, -C(=NH)N(C 1-6 alkyl)2, -C(=NH)NH(C 1-6 alkyl), -C(=NH)NH2, -OC(=NH)N(C 1-6 alkyl)2, -OC(NH)NH(C 1-6 alkyl), -OC(NH)NH2, -NHC(NH)N(C 1-6 alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1-6 alkyl), -SO2N(C 1-6 alkyl)2, -SO2NH(C 1-6 alkyl), -SO2NH2, -SO2C 1-6 alkyl, -SO2OC 1-6 alkyl, -OSO2C 1-6 alkyl, -SOC 1-6 alkyl, -Si(C 1-6 alkyl)3, -OSi(C 1-6 alkyl)3, -C(=S)N(C 1-6 alkyl)2, C(=S)NH(C 1-6 alkyl), C(=S)NH2, -C(=O)S(C 1-6 alkyl), -C(=S)SC 1-6 alkyl, -SC(=S)SC 1-6 alkyl, -P(=O)2(C 1-6 alkyl), -P(=O)(C 1-6 alkyl)2, -OP(=O)(C 1-6(alkyl)2, -OP(=O)(OC 1-6 (alkyl)2, C 1-6 alkyl, C 1-6 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, C6-C 10 aryl, C3-C7 heterocycloalkyl, C5-C 10 heteroaryl; or two geminal R gg substituents may combine to form =O or =S; wherein, X - is a counterion.

[0051] Exemplary substituents on the nitrogen atom include, but are not limited to: hydrogen, -OH, -OR aa (, -N(R cc )2, -CN, -C(=O)R aa (, -C(=O)N(R cc )2, -CO2R aa (, -SO2R aa (, -C(=NR bb )R aa (, -C(=NR cc )OR aa (, -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc (, -SO2OR cc (, -SOR aa (, -C(=S)N(R cc )2, -C(=O)SR cc (, -C(=S)SR cc (, -P(=O)2R aa (, -P(=O)(R aa )2, -P(=O)2N(R cc )2, -P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, or two R cc groups attached to the nitrogen atom combine to form a heterocycloalkyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 R dd groups, and wherein R aa , R bb , R cc and R dd are as described above.

[0052] Other definitions

[0053] As used herein, the term "pharmaceutically acceptable salt" refers to those carboxylate salts and amino acid addition salts of the compounds of the present invention that are suitable for contact with patient tissues within the scope of sound medical judgment, do not produce undue toxicity, irritation, allergic reactions, etc., are commensurate with a reasonable benefit / risk ratio, and are effective for their intended use, including (where possible) the zwitterionic forms of the compounds of the present invention.

[0054] "Subjects" to whom administration is made include, but are not limited to: humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults or elderly adults)) and / or non-human animals, e.g., mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats and / or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms "human", "patient" and "subject" may be used interchangeably herein.

[0055] "Disease", "disorder" and "condition" are used interchangeably herein.

[0056] Generally, an "effective amount" of a compound refers to an amount sufficient to elicit a desired biological response. As will be understood by those of ordinary skill in the art, the effective amount of the compounds of the present invention may vary depending on factors such as, for example, the biological target, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health status and symptoms of the subject. Effective amounts include therapeutically effective amounts and prophylactically effective amounts.

[0057] "Combination" and related terms refer to the administration of the compounds of the present invention and other therapeutic agents either simultaneously or sequentially. For example, the compounds of the present invention may be administered simultaneously or sequentially with other therapeutic agents in separate unit dosage forms, or simultaneously with other therapeutic agents in a single unit dosage form. Specific embodiments

[0058] As used herein, "compounds of the present invention" refers to compounds of formula (I) below (including sub-generic formulas, such as formula (II), formula (III), formula (IV), formula (V), etc.), their pharmaceutically acceptable salts, enantiomers, diastereomers, solvates, hydrates or isotopic variants, and mixtures thereof.

[0059] In one embodiment, the present invention relates to compounds of formula (I), or their pharmaceutically acceptable salts, isotopic variants, tautomers, stereoisomers, prodrugs, polymorphs, hydrates or solvates:

[0060]

[0061] Wherein,

[0062] R1 is selected from C 1-6 alkyl, C 1-6 haloalkyl, -(CH2) 1-4 -NH2, -(CH2) 1-4 -OH, C 3-7 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl, which may optionally be substituted by 1, 2, 3, 4 or 5 R*; R* is selected from halogen, CN, OH, NH2, C 1-6 alkyl, C 3-6 cycloalkyl or C 1-6 haloalkyl;

[0063] R2 is selected from CN, OH or NH2;

[0064] R3 is selected from H, -O-C 3-7 cycloalkyl or -O-3- to 7-membered heterocyclic group;

[0065] R4 is selected from H or -(CH2) 1-4 -3- to 7-membered heterocyclic group, and the -(CH2) 1-4 -3- to 7-membered heterocyclic group may optionally be substituted by 1, 2, 3, 4 or 5 substituents selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl;

[0066] R5 is selected from H, -C≡C-R 5a or -P(O)-(C 1-6 alkyl)2;

[0067] R 5a is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl, which may optionally be substituted by 1, 2, 3, 4 or 5 R#; R# is selected from halogen, CN, OH, NH2, C 1-6 alkyl or C 1-6 haloalkyl;

[0068] R6 is halogen;

[0069] R7 is selected from H, halogen or C 1-6 alkyl;

[0070] R8 is selected from H, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy or C 1-6 haloalkoxy.

[0071] R1

[0072] In one specific embodiment, R1 is C 1-6 alkyl; in another specific embodiment, R1 is C 1-4 alkyl; in another specific embodiment, R1 is C 1-2 alkyl; in another specific embodiment, R1 is C 1-6 haloalkyl; in another specific embodiment, R1 is C 1-4 haloalkyl; in another specific embodiment, R1 is -(CH2) 1-4 -NH2; in another specific embodiment, R1 is -(CH2) 1-3 -NH2; in another specific embodiment, R1 is -(CH2) 1-4 -OH; in another specific embodiment, R1 is -(CH2) 1-3 -OH; in another specific embodiment, R1 is C 3-7 cycloalkyl; in another specific embodiment, R1 is C 3-5 cycloalkyl; in another specific embodiment, R1 is a 3- to 10-membered heterocyclic group; in another specific embodiment, R1 is a 3- to 7-membered heterocyclic group; in another specific embodiment, R1 is C 6-10 aryl; in another specific embodiment, R1 is a 5- to 10-membered heteroaryl.

[0073] In one specific embodiment, R1 is -CH3; in another specific embodiment, R1 is cyclopropyl; in another specific embodiment, R1 is -CHF2; in another specific embodiment, R1 is -CH2CF3; in another specific embodiment, R1 is -CH2CH2OH; in another specific embodiment, R1 is In another specific embodiment, R1 is In another specific embodiment, R1 is In another specific embodiment, R1 is

[0074] In one specific embodiment, R1 is unsubstituted; in another specific embodiment, R1 is substituted with one R*; in another specific embodiment, R1 is substituted with two R*; in another specific embodiment, R1 is substituted with three R*; in another specific embodiment, R1 is substituted with four R*; in another specific embodiment, R1 is substituted with five R*.

[0075] R2

[0076] In one specific embodiment, R2 is CN; in another specific embodiment, R2 is OH; in another specific embodiment, R2 is NH2.

[0077] R3

[0078] In one specific embodiment, R3 is H; in another specific embodiment, R3 is -O-C 3-7 cycloalkyl; in another specific embodiment, R3 is -O-C 3-5 cycloalkyl; in another specific embodiment, R3 is -O-cyclopropyl; in another specific embodiment, R3 is -O-3- to 7-membered heterocyclic group;

[0079] R4

[0080] In one specific embodiment, R4 is H; in another specific embodiment, R4 is -(CH2) 1-4 -3- to 7-membered heterocyclic group.

[0081] In one specific embodiment, R4 is -(CH2) 1-4 -3- to 7-membered heterocyclic group, and the -(CH2) 1-4 -3- to 7-membered heterocyclic group is unsubstituted; in another specific embodiment, R4 is -(CH2) 1-4 -3- to 7-membered heterocyclic group, and the -(CH2) 1-4 -3- to 7-membered heterocyclic group is substituted by 1 substituent selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl; in another specific embodiment, R4 is -(CH2) 1-4 -3- to 7-membered heterocyclic group, and the -(CH2) 1-4 -3- to 7-membered heterocyclic group is substituted by 2 substituents selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl; in another specific embodiment, R4 is -(CH2) 1-4 -3- to 7-membered heterocyclic group, and the -(CH2) 1-4 -3- to 7-membered heterocyclic group is substituted by 3 substituents selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl; in another specific embodiment, R4 is -(CH2) 1-4 -3- to 7-membered heterocyclic group, and the -(CH2) 1-4 -3- to 7-membered heterocyclic group is substituted by 4 substituents selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl; in another specific embodiment, R4 is -(CH2) 1-4 -3- to 7-membered heterocyclic group, and the -(CH2)1-4 -3-7 membered heterocyclic group is substituted by 5 substituents selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl.

[0082] In one specific embodiment, R4 is

[0083] R5

[0084] In one specific embodiment, R5 is H; in another specific embodiment, R5 is -C≡C-R 5a ; in another specific embodiment, R5 is -P(O)-(C 1-6 alkyl)2; in another specific embodiment, R5 is -P(O)-(C 1-4 alkyl)2; in another specific embodiment, R5 is -P(O)-(CH3)2.

[0085] In one specific embodiment, R 5a is H; in another specific embodiment, R 5a is C 1-6 alkyl; in another specific embodiment, R 5a is C 1-4 alkyl; in another specific embodiment, R 5a is C 1-6 haloalkyl; in another specific embodiment, R 5a is C 1-4 haloalkyl; in another specific embodiment, R 5a is C 3-10 cycloalkyl; in another specific embodiment, R 5a is C 3-7 cycloalkyl; in another specific embodiment, R 5a is C 3-5 cycloalkyl; in another specific embodiment, R 5a is a 3- to 10-membered heterocyclic group; in another specific embodiment, R 5a is C 6-10 aryl; in another specific embodiment, R 5a is a 5- to 10-membered heteroaryl.

[0086] In one specific embodiment, R 5a is H; in another specific embodiment, R 5a is -CH3; in another specific embodiment, R 5a is -CF3; in another specific embodiment, R 5a is -C(CH3)2OH; in another specific embodiment, R5a is cyclopropyl.

[0087] In one specific embodiment, R 5a is unsubstituted; in another specific embodiment, R 5a is substituted by one R#; in another specific embodiment, R 5a is substituted by two R#; in another specific embodiment, R 5a is substituted by three R#; in another specific embodiment, R 5a is substituted by four R#; in another specific embodiment, R 5a is substituted by five R#.

[0088] R6

[0089] In one specific embodiment, R6 is a halogen, such as F.

[0090] R7

[0091] In one specific embodiment, R7 is H; in another specific embodiment, R7 is a halogen; in another specific embodiment, R7 is C 1-6 alkyl; in another specific embodiment, R7 is C 1-4 alkyl.

[0092] R8

[0093] In one specific embodiment, R8 is H; in another specific embodiment, R8 is a halogen, such as Cl; in another specific embodiment, R8 is C 1-6 alkyl; in another specific embodiment, R8 is C 1-4 alkyl, such as methyl; in another specific embodiment, R8 is C 1-6 haloalkyl; in another specific embodiment, R8 is C 1-4 haloalkyl; in another specific embodiment, R8 is C 1-6 alkoxy; in another specific embodiment, R8 is C 1-4 alkoxy, such as methoxy or ethoxy; in another specific embodiment, R8 is C 1-6 haloalkoxy; in another specific embodiment, R8 is C 1-4 haloalkoxy.

[0094] R*

[0095] In one specific embodiment, R* is a halogen; in another specific embodiment, R* is CN; in another specific embodiment, R* is OH; in another specific embodiment, R* is NH2; in another specific embodiment, R* is C 1-6alkyl; in another specific embodiment, R* is C 1-4 alkyl; in another specific embodiment, R* is C 1-6 haloalkyl; in another specific embodiment, R* is C 1-4 haloalkyl.

[0096] R#

[0097] In one specific embodiment, R# is halogen; in another specific embodiment, R# is CN, OH; in another specific embodiment, R# is NH2; in another specific embodiment, R# is C 1-6 alkyl; in another specific embodiment, R# is C 1-4 alkyl; in another specific embodiment, R# is C 1-6 haloalkyl; in another specific embodiment, R# is C 1-4 haloalkyl.

[0098] Any technical solution or any combination thereof in any of the above specific embodiments can be combined with any technical solution or any combination thereof in other specific embodiments. For example, any technical solution or any combination thereof of L can be combined with any technical solution or any combination thereof of R1-R8, R*, R#, etc. The present invention aims to include all such combinations of technical solutions, and for the sake of brevity, they are not listed one by one here.

[0099] In a more specific embodiment, the present invention provides the compound of formula (I) above, or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0100] R1 is selected from C 1-4 alkyl, C 1-4 haloalkyl, -(CH2) 1-3 -NH2, -(CH2) 1-3 -OH, C 3-5 cycloalkyl or a 3- to 7-membered heterocyclic group, which may optionally be substituted by one, two or three R*; R* is selected from halogen or C 1-4 alkyl;

[0101] R2 is selected from CN, OH or NH2;

[0102] R3 is selected from H or -O-C 3-5 cycloalkyl; preferably -O-C 3-5 cycloalkyl;

[0103] R4 is selected from H or -(CH2) 1-3 -3- to 7-membered heterocyclic group, the said -(CH2) 1-3-3-7-membered heterocyclic group may optionally be substituted with 1, 2 or 3 C 1-4 alkyl groups; preferably H;

[0104] R5 is selected from H, -C≡C-R 5a or -P(O)-(C 1-4 alkyl)2; preferably -C≡C-R 5a ;

[0105] R 5a is selected from H, C 1-4 alkyl, C 1-4 haloalkyl or C 3-5 cycloalkyl, which may optionally be substituted with 1, 2 or 3 R#; R# is selected from OH or C 1-4 alkyl;

[0106] R6 is a halogen;

[0107] R7 is selected from H, a halogen or C 1-4 alkyl;

[0108] R8 is H, a halogen, C 1-4 alkyl or C 1-4 alkoxy.

[0109] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0110] R1 is selected from -CH3, cyclopropyl, -CH2CF3, -CH2CH2OH,

[0111] R2 is CN;

[0112] R3 is selected from H or -O-cyclopropyl; preferably -O-cyclopropyl;

[0113] R4 is selected from H or preferably H;

[0114] R5 is selected from H, -C≡C-R 5a or -P(O)-(CH3)2; preferably -C≡C-R 5a ;

[0115] R 5a is selected from H, -CH3, -CF3, -C(CH3)2OH or cyclopropyl;

[0116] R6 is F;

[0117] R7 is H;

[0118] R8 is H, Cl, methyl, methoxy or ethoxy.

[0119] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, which has the following structure:

[0120]

[0121] wherein each group is as defined above.

[0122] In a more specific embodiment, the present invention provides a compound of formula (II), or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

[0123]

[0124] wherein,

[0125] R1 is selected from C 1-6 alkyl or C 1-6 haloalkyl;

[0126] R3 is selected from -O-C 3-7 cycloalkyl or -O-3-7-membered heterocyclic group;

[0127] R 5a is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, C 3-10 cycloalkyl, 3-10-membered heterocyclic group, C 6-10 aryl or 5-10-membered heteroaryl, which may optionally be substituted by 1, 2, 3, 4 or 5 R#; R# is selected from halogen, CN, OH, NH2, C 1-6 alkyl or C 1-6 haloalkyl;

[0128] R6 is halogen;

[0129] R7 is selected from H, halogen or C 1-6 alkyl.

[0130] In a more specific embodiment, the present invention provides a compound of formula (II) above, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0131] R1 is C 1-6 alkyl;

[0132] R3 is -O-C 3-5 cycloalkyl;

[0133] R5a Selected from H, C 1-6 alkyl, C 1-6 haloalkyl or C 3-7 cycloalkyl, which may optionally be substituted by one, two or three R#; R# is selected from halogen, OH, NH2, C 1-6 alkyl or C 1-6 haloalkyl;

[0134] R6 is halogen;

[0135] R7 is selected from H, halogen or C 1-4 alkyl.

[0136] In a more specific embodiment, the present invention provides a compound of formula (III), or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

[0137]

[0138] wherein,

[0139] R1 is C 1-4 alkyl;

[0140] R 5a is selected from H, C 1-4 alkyl, C 1-4 haloalkyl or C 3-5 cycloalkyl, which may optionally be substituted by one, two or three R#; R# is selected from OH or C 1-4 alkyl;

[0141] R6 is halogen.

[0142] In a more specific embodiment, the present invention provides the above compound of formula (III), or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0143] R1 is C 1-2 alkyl, preferably -CH3;

[0144] R 5a is selected from H, -CH3, -CF3, -C(CH3)2OH or cyclopropyl;

[0145] R6 is halogen, preferably F.

[0146] In a more specific embodiment, the present invention provides a compound of formula (II), or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

[0147]

[0148] Among them,

[0149] R1 is selected from C 1-6 alkyl, C 1-6 haloalkyl, -(CH2) 1-4 -NH2, -(CH2) 1-4 -OH, C 3-7 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl, which may optionally be substituted by 1, 2, 3, 4 or 5 R*; R* is selected from halogen, CN, OH, NH2, C 1-6 alkyl or C 1-6 haloalkyl;

[0150] R3 is -O-C 3-7 cycloalkyl;

[0151] R 5a is selected from C 1-6 alkyl or C 1-6 haloalkyl;

[0152] R6 is halogen;

[0153] R7 is selected from H, halogen or C 1-6 alkyl.

[0154] In a more specific embodiment, the present invention provides the compound of formula (II) above, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0155] R1 is selected from C 1-6 alkyl, C 1-6 haloalkyl, -(CH2) 1-4 -NH2, -(CH2) 1-4 -OH, C 3-7 cycloalkyl or 3- to 10-membered heterocyclic group, which may optionally be substituted by 1, 2 or 3 R*; R* is selected from halogen, C 1-6 alkyl or C 1-6 haloalkyl;

[0156] R3 is -O-C 3-5 cycloalkyl;

[0157] R 5a is C 1-6 alkyl or C 1-6 haloalkyl;

[0158] R6 is halogen;

[0159] R7 is selected from H, halogen or C1-4 Alkyl

[0160] In a more specific embodiment, the present invention provides a compound of formula (III), or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

[0161]

[0162] Wherein,

[0163] R1 is selected from C 1-4 alkyl, C 1-4 haloalkyl, -(CH2) 1-3 -NH2, -(CH2) 1-3 -OH, C 3-5 cycloalkyl or a 3- to 7-membered heterocyclic group, which may optionally be substituted by 1, 2 or 3 R*; R* is selected from halogen or C 1-4 alkyl;

[0164] R 5a is C 1-4 alkyl or C 1-4 haloalkyl;

[0165] R6 is halogen.

[0166] In a more specific embodiment, the present invention provides the above compound of formula (III), or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0167] R1 is selected from -CH3, -CHF2, -CH2CF3, -CH2CH2OH, cyclopropyl,

[0168] R 5a is C 1-2 alkyl, preferably -CH3;

[0169] R6 is halogen, preferably F.

[0170] In a more specific embodiment, the present invention provides a compound of formula (IV), or a pharmaceutically acceptable salt, isotope variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

[0171]

[0172] Wherein,

[0173] R1 is selected from C 1-6 alkyl or C 1-6 haloalkyl;

[0174] R3 is selected from -O-C 3-7 cycloalkyl or -O-3- to 7-membered heterocyclic group;

[0175] R 5a is selected from C 1-6 alkyl or C 1-6 haloalkyl;

[0176] R6 is halogen;

[0177] R7 is selected from H, halogen or C 1-6 alkyl;

[0178] R8 is selected from H, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy or C 1-6 haloalkoxy;

[0179] In a more specific embodiment, the present invention provides a compound of formula (IV) above, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0180] R1 is C 1-6 alkyl;

[0181] R 5a is C 1-6 alkyl;

[0182] R3 is -O-C 3-7 cycloalkyl, preferably -O-C 3-5 cycloalkyl;

[0183] R6 is halogen;

[0184] R7 is selected from H, halogen or C 1-4 alkyl;

[0185] R8 is selected from halogen, C 1-6 alkyl or C 1-6 alkoxy, preferably halogen or C 1-6 alkyl;

[0186] In a more specific embodiment, the present invention provides a compound of formula (V), or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

[0187]

[0188] wherein,

[0189] R1 is C 1-4 alkyl;

[0190] R 5a is C 1-4 alkyl;

[0191] R6 is halogen;

[0192] R8 is selected from halogen, C 1-4 alkyl or C 1-4 alkoxy, preferably halogen or C 1-4 alkyl;

[0193] In a more specific embodiment, the present invention provides a compound of formula (V) above, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof, wherein,

[0194] R1 is C 1-2 alkyl, preferably methyl;

[0195] R 5a is C 1-2 alkyl, preferably methyl;

[0196] R6 is halogen, preferably F;

[0197] R8 is selected from halogen, C 1-2 alkyl or C 1-2 alkoxy, preferably halogen or C 1-2 alkyl, more preferably Cl or methyl.

[0198] In a more specific embodiment, the present invention provides a compound, or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from:

[0199]

[0200] The compounds of the present invention may include one or more asymmetric centers and, therefore, may exist in various stereoisomeric forms, for example, enantiomeric and / or diastereomeric forms. For example, the compounds of the present invention may be individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. The isomers can be separated from the mixture by methods known to those skilled in the art, including: chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or the preferred isomers can be prepared by asymmetric synthesis.

[0201] The compounds of the present invention may also exist as tautomers. For compounds that exist in different tautomeric forms, a given compound is not limited to any particular tautomeric form, but is intended to encompass all tautomeric forms.

[0202] Those skilled in the art will understand that organic compounds can form complexes with solvents, which react in the solvent or precipitate or crystallize out from the solvent. These complexes are called "solvates". When the solvent is water, the complex is called a "hydrate". The present invention encompasses all solvates of the compounds of the present invention.

[0203] The term "solvate" refers to a form of a compound or its salt that is combined with a solvent, usually formed by a solvolysis reaction. This physical association may include hydrogen bonding. Conventional solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, etc. The compounds described herein can be prepared, for example, in crystalline form and can be solvated. Suitable solvates include pharmaceutically acceptable solvates and further include stoichiometric solvates and non-stoichiometric solvates. In some cases, the solvate will be capable of being isolated, for example, when one or more solvent molecules are incorporated into the lattice of the crystalline solid. "Solvate" includes solvates in solution state and isolable solvates. Representative solvates include hydrates, ethanolates, and methanolates.

[0204] The term "hydrate" refers to a compound combined with water. Generally, the ratio of the number of water molecules contained in the hydrate of a compound to the number of molecules of that compound in the hydrate is determined. Thus, a hydrate of a compound can be represented, for example, by the general formula R·xH2O, where R is the compound and x is a number greater than 0. A given compound can form more than one type of hydrate, including, for example, monohydrates (x is 1), lower hydrates (x is a number greater than 0 and less than 1, e.g., hemihydrate (R·0.5H2O)), and polyhydrates (x is a number greater than 1, e.g., dihydrate (R·2H2O) and hexahydrate (R·6H2O)).

[0205] The compounds of the present invention can be in amorphous or crystalline form (polymorphs). In addition, the compounds of the present invention can exist in one or more crystalline forms. Thus, the present invention includes within its scope all amorphous or crystalline forms of the compounds of the present invention. The term "polymorph" refers to a crystalline form (or its salt, hydrate, or solvate) of a compound with a specific crystal packing arrangement. All polymorphs have the same elemental composition. Different crystalline forms usually have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, photoelectric properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, and other factors can result in one crystalline form being dominant. The various polymorphs of a compound can be prepared by crystallization under different conditions.

[0206] The present invention also includes isotopically labeled compounds (isotope variants) which are equivalent to those described by formula (I), but in which one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number commonly found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, for example 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F and 36 Cl. The compounds of the present invention containing the above isotopes and / or other isotopes of other atoms, their prodrugs, and the pharmaceutically acceptable salts of said compounds or said prodrugs are all within the scope of the present invention. Certain isotopically labeled compounds of the present invention, for example those incorporating radioactive isotopes (e.g. 3 H and 14 C), can be used for drug and / or substrate tissue distribution assays. Tritium, i.e. 3 H and carbon-14, i.e. 14 C isotopes are particularly preferred because they are readily prepared and detected. Furthermore, replacement with heavier isotopes, such as deuterium, i.e. 2 H, can provide therapeutic benefits such as an extended in vivo half-life or a reduced dosage requirement due to higher metabolic stability, and may thus be preferred in some cases. The isotopically labeled compounds of formula (I) of the present invention and their prodrugs can generally be prepared by using readily available isotopically labeled reagents in place of non-isotopically labeled reagents when carrying out the processes and / or the procedures and preparation examples disclosed below.

[0207] In addition, prodrugs are also included within the context of the present invention. As used herein, the term "prodrug" refers to a compound that is converted in vivo, for example, by hydrolysis in the blood, into its active form that has a medical effect. Pharmaceutically acceptable prodrugs are described in T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, and D. Fleisher, S. Ramon, and H. Barbra, "Improved oral drug delivery: solubility limitations overcome by the use of prodrugs", Advanced Drug Delivery Reviews (1996) 19(2) 115-130, each incorporated herein by reference.

[0208] Pharmaceutical Compositions and Kits

[0209] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention (also referred to as the "active ingredient") and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises an effective amount of a compound of the present invention. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of a compound of the present invention. In some embodiments, the pharmaceutical composition comprises a prophylactically effective amount of a compound of the present invention.

[0210] A pharmaceutically acceptable excipient for use in the present invention refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compounds formulated together. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present invention include (but are not limited to) ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffering substances (such as phosphates), glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polypropylene-block polymers, polyethylene glycol, and lanolin.

[0211] The present invention also includes a kit (e.g., a pharmaceutical package). The provided kit may include a compound of the present invention, other therapeutic agents, and first and second containers (e.g., vials, ampoules, bottles, syringes, and / or dispersible packages or other suitable containers) containing the compound of the present invention and other therapeutic agents. In some embodiments, the provided kit may also optionally include a third container containing a pharmaceutical excipient for diluting or suspending the compound of the present invention and / or other therapeutic agents. In some embodiments, the combination of the compound of the present invention and other therapeutic agents provided in the first and second containers forms a unit dosage form.

[0212] Administration

[0213] The pharmaceutical compositions provided by the present invention can be administered by many routes, including but not limited to: oral administration, parenteral administration, inhalation administration, topical administration, rectal administration, nasal administration, buccal administration, vaginal administration, administration by implant, or other administration methods. For example, parenteral administration as used herein includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, intra-articular administration, intra-arterial administration, intra-synovial cavity administration, intrasternal administration, intrathecal administration, intralesional administration, and intracranial injection or infusion techniques.

[0214] Generally, an effective amount of the compound provided herein is administered. Depending on the circumstances, including the disorder being treated, the route of administration selected, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, etc., the amount of the compound actually administered can be determined by a physician.

[0215] When used to prevent the disorders described herein, the compound provided herein is administered to a subject at risk of developing the disorder, typically on the advice of a physician and under the physician's supervision, at a dosage level as described above. Subjects at risk of developing a specific disorder generally include those with a family history of the disorder, or those determined to be particularly sensitive to developing the disorder by genetic testing or screening.

[0216] The pharmaceutical compositions provided herein can also be administered over a long period of time (“chronic administration”). Chronic administration refers to the administration of a compound or its pharmaceutical composition over a long period of time, e.g., 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or can be administered indefinitely, e.g., for the remainder of the subject's life. In some embodiments, chronic administration is intended to provide a constant level of the compound in the blood over a long period of time, e.g., within a therapeutic window.

[0217] A variety of administration methods can be used to further deliver the pharmaceutical compositions of the present invention. For example, in some embodiments, the pharmaceutical composition can be administered by bolus injection, for example, to increase the concentration of the compound in the blood to an effective level. The bolus dose depends on the target systemic level of the active ingredient through the body. For example, an intramuscular or subcutaneous bolus dose results in a slow release of the active ingredient, while a bolus directly delivered to a vein (e.g., by IV infusion) enables a more rapid delivery, causing the concentration of the active ingredient in the blood to quickly rise to an effective level. In other embodiments, the pharmaceutical composition can be administered in the form of a continuous infusion, for example, by IV infusion, to provide a steady-state concentration of the active ingredient in the body of the subject. Additionally, in other embodiments, a bolus dose of the pharmaceutical composition can be administered first, followed by a continuous infusion.

[0218] Oral compositions can be in the form of bulk liquid solutions or suspensions or bulk powders. However, more commonly, the compositions are provided in unit dosage form for ease of accurate dosing. The term "unit dosage form" refers to physically discrete units suitable as unit doses for human patients and other mammals, each unit containing a predetermined quantity of the active substance, which is suitable for producing the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampoules or syringes of liquid compositions, or pills, tablets, capsules, etc. in the case of solid compositions. In such compositions, the compound is typically the minor component (from about 0.1 to about 50% by weight, or preferably from about 1 to about 40% by weight), with the remainder being various carriers or excipients and processing aids useful for forming the desired dosage form.

[0219] For oral dosing, a representative regimen is one to five oral doses per day, especially two to four oral doses, typically three oral doses. Using these dosing patterns, each dose provides from about 0.01 to about 20 mg / kg of the compound of the present invention, with preferred doses each providing from about 0.1 to about 10 mg / kg, especially from about 1 to about 5 mg / kg.

[0220] To provide blood levels similar to or lower than those obtained with injection doses, transdermal doses are typically selected in an amount of from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight.

[0221] From about 1 to about 120 hours, particularly 24 to 96 hours, the infusion dosage level ranges from about 0.1 mg / kg / hour to at least 10 mg / kg / hour. To achieve a sufficient steady-state level, a loading bolus of about 0.1 mg / kg to about 10 mg / kg or more may also be administered. For a human patient weighing 40 to 80 kg, the maximum total dose should not exceed about 2 g / day.

[0222] Liquid forms suitable for oral administration may include suitable aqueous or non-aqueous carriers, as well as buffering agents, suspending and dispersing agents, coloring agents, flavoring agents, and the like. Solid forms may include, for example, any of the following components, or compounds with similar properties: binders, such as microcrystalline cellulose, tragacanth, or gelatin; excipients, such as starch or lactose; disintegrants, such as alginic acid, Primogel, or corn starch; lubricants, such as magnesium stearate; glidants, such as colloidal silicon dioxide; sweeteners, such as sucrose or saccharin; or flavoring agents, such as peppermint, methyl salicylate, or orange flavoring.

[0223] Injectable compositions are typically based on injectable sterile saline or phosphate-buffered saline, or other injectable excipients known in the art. As previously mentioned, in such compositions, the active compound is typically the minor component, often about 0.05 to 10% by weight, with the remainder being injectable excipients and the like.

[0224] Transdermal compositions are typically formulated as topical ointments or creams containing the active component. When formulated as an ointment, the active component is typically combined with paraffin or a water-miscible ointment base. Alternatively, the active component may be formulated as a cream with, for example, an oil-in-water cream base. Such transdermal preparations are well known in the art and typically include other components for enhancing the steady skin penetration of the active component or the preparation. All such known transdermal preparations and components are included within the scope provided by the present invention.

[0225] The compounds of the present invention may also be administered by transdermal devices. Thus, transdermal administration may be achieved using patches of the reservoir or porous membrane type, or various solid matrices.

[0226] The above components of the compositions for oral, injectable, or topical administration are merely representative. Other materials, as well as processing techniques, etc., are described in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, which is hereby incorporated by reference.

[0227] The compounds of the present invention can also be administered in a sustained release form or from a sustained release delivery system. Descriptions of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

[0228] The present invention also relates to pharmaceutically acceptable formulations of the compounds of the present invention. In one embodiment, the formulation comprises water. In another embodiment, the formulation comprises a cyclodextrin derivative. The most common cyclodextrins are α-, β- and γ-cyclodextrins, which are composed of 6, 7 and 8 α-1,4-linked glucose units, respectively, and optionally include one or more substituents on the linked sugar moieties, including but not limited to: methylated, hydroxyalkylated, acylated and sulfonated alkyl ether substitutions. In some embodiments, the cyclodextrin is sulfonated alkyl ether β-cyclodextrin, for example, sulfobutyl ether β-cyclodextrin, also known as Captisol. See, e.g., U.S. 5,376,645. In some embodiments, the formulation comprises hexapropyl-β-cyclodextrin (e.g., in water, 10-50%).

[0229] Examples

[0230] The reagents used in the present invention are commercially available reagents purchased directly or synthesized by common methods well known in the art.

[0231] Notes on common abbreviations:

[0232] PE = petroleum ether; EA = ethyl acetate; MeOH = methanol; DCM = dichloromethane; DCE = dichloroethane; CH3CN = acetonitrile; 1,4-dioxane = 1,4-dioxane; DMSO = dimethyl sulfoxide; HFIP = hexafluoroisopropanol; DMF = N,N-dimethylformamide; Hex = n-hexane; IPA = isopropanol; NMP = N-methylpyrrolidone; NMO = N-methylmorpholine-N-oxide; TEA = triethylamine; DIEA = diisopropylethylamine; CuI = copper iodide; CuCN = copper cyanide; triphosgene = triphosgene; p-TsOH = p-toluenesulfonic acid.

[0233] The specific reaction routes or steps of the following examples are used in the present invention, specifically as follows:

[0234] Example 1

[0235] Preparation of key intermediates

[0236] Synthesis of intermediates a1, a6-a8

[0237]

[0238] Step 1: Under nitrogen protection, in an ice bath, dissolve NaH (1.27 g, 54.0 mmol) in 50 mL of anhydrous tetrahydrofuran. Slowly add cyclopropyl alcohol (4.35 g, 63.6 mmol). After stirring for 1 hour, add starting material a1-1 (8.5 g, 49.1 mmol). Heat to 60 °C and react for 2 hours. Monitor the reaction completion by LC-MS. Stop the reaction, add 100 mL of water to the system, extract with ethyl acetate. Combine the organic phases, dry over anhydrous sodium sulfate, and separate by column chromatography (PE / EA, 9 / 1) to obtain a pale yellow solid a1-2 (2.5 g), yield: 24%.

[0239] Step 2: At -78 °C, under nitrogen protection, dissolve the intermediate a1-2 from the previous step (2.5 g, 11.8 mmol) in 50 mL of anhydrous tetrahydrofuran. Slowly add lithium diisopropylamide LDA (17.7 mmol, 2 M). After stirring for 0.5 hour, add elemental iodine (6.0 g, 23.6 mmol). Slowly warm to room temperature and react for 12 hours. Monitor the reaction completion by LC-MS. Add 100 mL of water to the system, extract with ethyl acetate. Combine the organic phases, dry over anhydrous sodium sulfate, and separate by column chromatography (PE / EA, 9 / 1) to obtain a white solid intermediate a1-3 (3.6 g), yield: 90%.

[0240] Step 3: Under nitrogen protection, dissolve the intermediate a1-3 from the previous step (3.6 g, 10.6 mmol), starting material N-1-methyl-5-pyrazoleboronic acid pinacol ester a1-4 (6.6 g, 31.8 mmol) and Na2CO3 (2.2 g, 21.2 mmol) in a mixed solution of 15 mL of 1,4-dioxane and water (v / v, 5 / 1). Add Pd(dtbpf)Cl2 (684 mg, 1.1 mmol). Heat to 80 °C and react for 12 hours. Cool to room temperature, add water to the system, extract with ethyl acetate. Combine the organic phases, dry over anhydrous sodium sulfate, and separate by column chromatography (PE / EA, 5 / 1) to obtain a pale yellow solid a1 (1.4 g), yield: 45%. LC-MS: [M+H] + = 292.

[0241] Refer to the synthetic route of intermediate a1 to synthesize the following intermediates.

[0242]

[0243] Synthesis of intermediates a2, a3, a9 - a11

[0244]

[0245] Procedure: Under ice bath and nitrogen protection, dissolve intermediate a1 (1.4 g, 4.8 mmol) and N-iodosuccinimide NIS (1.08 g, 4.8 mmol) in 20 mL of glacial acetic acid, heat to 80 °C and react for 3 hours, then cool to room temperature. Add 100 mL of ice water to the system, extract with ethyl acetate, wash the organic phase with saturated aqueous sodium bicarbonate solution, dry over anhydrous sodium sulfate, concentrate, and separate the crude product by column chromatography (PE / EA, 4 / 1) to obtain intermediate a2 (1.3 g), yield: 65%. LC-MS: [M+H] + = 417.

[0246] Refer to the synthetic route of intermediate a2 to synthesize the following intermediates.

[0247]

[0248]

[0249] Synthesis of intermediate a4

[0250]

[0251] Procedure: Under nitrogen protection, dissolve raw material a4-1 (1.0 g, 2.83 mmol), bis(pinacolato)diboron B2Pin2 (1.44 g, 5.66 mmol) and KOAc (555 mg, 5.66 mmol) in 15 mL of 1,4-dioxane, add Pd(dppf)Cl2 (205 mg, 0.28 mmol), heat to 100 °C and react for 2 hours. Cool to room temperature, add water to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by flash column chromatography (PE / EA, 2 / 1) to obtain yellow solid a4 (970 mg), yield: 85%. LC-MS: [M+H] + = 402.

[0252] Synthesis of intermediate a5

[0253]

[0254] Step 1: Dissolve 3-fluoro-5-cyanobenzaldehyde a5-1 (4.0 g, 26.8 mmol) and N-methylpiperazine a5-2 (4.0 g, 40.2 mmol) in 80 mL of tetrahydrofuran. Add sodium triacetoxyborohydride (17.1 g, 80.5 mmol) and 0.5 mL of acetic acid, and react at room temperature for 12 hours. After detecting the completion of the reaction by LC-MS, add 30 mL of ice water to the system, evaporate the organic solvent under reduced pressure, extract with dichloromethane, dry over anhydrous sodium sulfate, concentrate, and separate by column chromatography (DCM / MeOH, 10 / 1) to obtain a yellow oil a5-3 (6.0 g), yield: 96%. LC-MS: [M+H] + = 234.

[0255] Step 2: Under nitrogen protection, at -78 °C, dissolve the intermediate a5-3 (4.0 g, 17.2 mmol) obtained in the previous step in 80 mL of tetrahydrofuran, slowly dropwise add LDA (5.51 g, 51.4 mmol), and stir for 2 hours. Dissolve elemental iodine (8.7 g, 34.3 mmol) in 20 mL of tetrahydrofuran, and gradually add it dropwise to the reaction system. Slowly warm up to room temperature and continue stirring for 12 hours. Quench the reaction by adding saturated ammonium chloride aqueous solution to the reaction solution, extract with dichloromethane, dry over anhydrous sodium sulfate, concentrate, and separate by column chromatography (DCM / MeOH, 10 / 1) to obtain a gray solid a5-4 (5.6 g), yield: 90%. LC-MS: [M+H] + = 360.

[0256] Step 3: Dissolve the intermediate a5-4 (2.0 g, 5.6 mmol), Na2CO3 (1.8 g, 16.8 mmol), and raw material a1-4 (3.48 g, 16.7 mmol) in a mixed solution of 50 mL of 1,4-dioxane and water (v / v, 9 / 1), and add the catalyst Pd(dtbpf)Cl2 (360 mg, 0.56 mmol). Heat up to 80 °C and react for 4 hours, then stop the reaction. Filter, evaporate the solvent under reduced pressure, and separate by column chromatography (DCM / MeOH, 10 / 1) to obtain a red solid a5-5 (1.2 g), yield: 68%. LC-MS: [M+H] + = 314.

[0257] Step 4: In an ice bath, dissolve the intermediate a5-5 (500 mg, 1.47 mmol) obtained in the previous step in 8 mL of chloroform, slowly dropwise add liquid bromine (720 mg, 4.5 mmol), continue the reaction for 3 hours, and then stop the reaction. Quench the reaction by adding saturated sodium thiosulfate aqueous solution to the system, extract with dichloromethane, dry the organic phase over anhydrous sodium sulfate, concentrate, and separate by column chromatography (DCM / MeOH, 8 / 1) to obtain the intermediate a5 (220 mg), yield: 38%. LC-MS: [M+H] += 392.

[0258] Synthesis of Intermediate b1 - b4

[0259]

[0260] Step 1: Dissolve 5 - bromo - 7 - chloro - isobenzofuran - 1(3H) - one b1 - 1 (9.5 g, 38.4 mmol) and tert - butoxybis(dimethylamino)methane b1 - 2 (8.0 g, 46.1 mmol) in 95 mL of toluene. Heat the mixture to 105 °C and react for 3 hours. Monitor the reaction by LC - MS until completion, then cool to room temperature. Evaporate the solvent under reduced pressure. Add 25 mL of methyl tert - butyl ether to the mixture, precipitate the solid, filter by suction, and dry the filter cake to obtain a yellow solid b1 - 3 (6.3 g), yield: 54%. LC - MS: [M + H] + = 302.

[0261] Step 2: Under nitrogen protection, dissolve the intermediate b1 - 3 (6.3 g, 20.7 mmol) obtained in the previous step in 126 mL of ethanol. Slowly add hydrazine hydrate (4.1 g, 82.6 mmol), heat the mixture to 95 °C and react for 48 hours, then cool to room temperature. Evaporate the solvent under reduced pressure. Add 25 mL of methyl tert - butyl ether to the mixture, precipitate the solid, filter by suction, and dry the filter cake to obtain a yellow solid b1 - 4 (5.2 g), yield: 80%. LC - MS: [M + H] + = 316.

[0262] Step 3: Under nitrogen protection, dissolve the intermediate b1 - 4 (5.2 g, 16.4 mmol) obtained in the previous step in 104 mL of tetrahydrofuran. Add isobutyl chloroformate b1 - 5 (2.7 g, 19.7 mmol), react at room temperature for 16 hours, and then stop the reaction. Evaporate the solvent under reduced pressure. Add 100 mL of water to the reaction solution, extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate, and separate by flash column chromatography (PE / EA, 5 / 1) to obtain a gray solid b1 - 6 (2.4 g), yield: 48%. LC - MS: [M + H] + = 306.

[0263] Step 4: Under ice - bath and nitrogen protection, dissolve bis(tert - butoxycarbonyl)amine NH(Boc)2 (1.7 g, 7.8 mmol) in 46 mL of anhydrous tetrahydrofuran. Slowly add LiHMDS (8.3 mL, 1 M), and stir at ice - bath temperature for 0.5 hour. Add the intermediate b1 - 6 (2.3 g, 7.5 mmol) obtained in the previous step to the reaction solution, heat to room temperature and react for 2 hours, then stop the reaction. Add 100 mL of saturated ammonium chloride aqueous solution to the system to quench the reaction, extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate, and obtain the intermediate b1 - 7 (2.1 g), yield: 55%. LC - MS: [M + H]+ = 489.

[0264] Step 5: Under nitrogen protection, dissolve the intermediate b1-7 (2.1 g, 4.3 mmol) from the previous step in 42 mL of acetonitrile, slowly add magnesium perchlorate (200 mg, 0.86 mmol), raise the temperature to 50 °C and react for 3 hours, then stop the reaction. Evaporate the solvent under reduced pressure, perform suction filtration, wash the filter cake with water and acetonitrile respectively, and dry to obtain a gray solid b1-8 (1.2 g), yield: 76%. LC-MS: [M+H] + = 389.

[0265] Step 6: Under nitrogen protection, dissolve the intermediate b1-8 (800 mg, 2.06 mmol), KOAc (606 mg, 6.2 mmol) and bis(pinacolato)diboron (784 mg, 3.1 mmol) in 16 mL of 1,4-dioxane, add the catalyst Pd(dppf)Cl2 (75 mg, 0.1 mmol). Raise the temperature to 10 °C and react for 1 hour, then stop the reaction. Filter, wash the filter cake with petroleum ether to obtain a gray solid b1 (665 mg), yield: 68%. LC-MS: [M+H] + = 436.

[0266] Refer to the synthetic route of intermediate b1 to synthesize the following intermediate.

[0267]

[0268] Example 2: Synthesis of control molecule M0

[0269]

[0270] Step 1: Under nitrogen protection, dissolve intermediate a2 (1.3 g, 3.1 mmol), intermediate a4 (1.5 g, 3.7 mmol) and K2CO3 (2.4 g, 18.6 mmol) in a mixed solution of 50 mL of 1,4-dioxane and water (v / v, 9 / 1), add RuPhosPd-G3 (251 mg, 0.3 mmol), raise the temperature to 60 °C and react for 12 hours. Cool to room temperature, add water to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by column chromatography (DCM / MeOH, 20 / 1) to obtain a pale yellow oil M0-1 (410 mg), yield: 23%. LC-MS: [M+H] + = 565.

[0271] Step 2: Dissolve the oily substance M0-1 (165 mg, 0.4 mmol) from the previous step in 5 mL of dichloromethane, add 1 mL of trifluoroacetic acid, stir at room temperature for 0.5 h, and detect by LCMS that the reaction is complete. Evaporate the solvent under reduced pressure, add 50 mL of saturated aqueous NaHCO3 to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by column chromatography (DCM / MeOH, 10 / 1) to obtain a pale yellow solid M0 (45 mg), yield: 33%, LC-MS: [M+H] + = 465.

[0272] 1H NMR (400 MHz, DMSO-d6) δ 12.61 (s, 1H), 8.32 (s, 1H), 8.17–8.19 (d, J = 8.4 Hz, 1H), 8.03–8.05 (d, J = 6.0 Hz, 1H), 7.80 (s, 1H), 7.65 - 7.67 (d, J = 8.4 Hz, 1H), 4.20–4.23 (dd, J = 5.8, 3.2 Hz, 1H), 3.99 (s, 2H), 3.81 (s, 3H), 0.97–0.91 (m, 2H), 0.84–0.85 (m, 2H).

[0273] Step 3: The compound M0 is separated by SFC chiral column chromatography to obtain the target molecules M0-a and M0-b.

[0274] Example 3: Synthesis of the target molecule P1

[0275]

[0276] Step 1: Under nitrogen protection, dissolve intermediate a5 (200 mg, 0.48 mmol), intermediate a4 (577 mg, 1.4 mmol) and K2CO3 (200 mg, 1.44 mmol) in a mixed solution of 10 mL of 1,4-dioxane and water (v / v, 9 / 1), add Pd(dppf)Cl2 (32 mg, 0.05 mmol), and react at 100 °C for 4 h. Cool to room temperature, add water to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by column chromatography (DCM / MeOH, 8 / 1) to obtain a gray solid P1-1 (110 mg), yield: 39%. LC-MS: [M+H] + = 587.

[0277] Step 2: Dissolve the oily substance P-1 (110 mg, 0.19 mmol) from the previous step in 5 mL of dichloromethane, add 1 mL of trifluoroacetic acid, stir at room temperature for 0.5 h, and detect by LCMS that the reaction is complete. Evaporate the solvent under reduced pressure, add 40 mL of saturated aqueous NaHCO3 to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by flash reverse column chromatography (CH3CN / H2O) to obtain white solid P1 (70 mg), yield: 76%, LC-MS: [M+H] + = 487.

[0278] 1 1H NMR (400 MHz, DMSO-d6) δ 12.87 (s, 1H), 8.43 (s, 2H), 8.35 (s, 1H), 8.11 (d, J = 8.3 Hz, 1H), 7.91 (s, 1H), 7.83 (d, J = 8.4 Hz, 2H), 7.41–7.36 (m, 1H), 4.35 (s, 2H), 3.76 (d, J = 5.3 Hz, 5H), 3.42 (s, 2H), 3.11 (d, J = 8.4 Hz, 2H), 2.99 (d, J = 8.4 Hz, 2H), 2.81 (s, 3H), 2.49–2.40 (m, 2H).

[0279] Example 4: Synthesis of the target molecule P2

[0280]

[0281] Step 1: Under nitrogen protection, dissolve compound M0-1 (100 mg, 0.18 mmol), dimethylphosphine oxide (16 mg, 0.2 mmol) and K3PO4 (42 mg, 0.2 mmol) in 8 mL of DMF, add the catalyst Pd(OAc)2 (3 mg, 0.009 mmol) and Xantphos (6.3 mg, 0.011 mmol), heat to 130 °C and react for 16 h, then cool to room temperature. Add water to the system, extract with ethyl acetate, dry over anhydrous sodium sulfate, and concentrate. The crude product is separated by flash column chromatography (DCM / MeOH, 10 / 1) to obtain compound P2-1 (20 mg), yield: 18%. LC-MS: [M+H] + = 607.

[0282] Step 2: Dissolve the intermediate P2-1 (20 mg, 0.033 mmol) from the previous step in 3 mL of dichloromethane, add 0.5 mL of trifluoroacetic acid, stir at room temperature for 0.5 h, and monitor the reaction completion by LCMS. Evaporate the solvent under reduced pressure, add 40 mL of saturated aqueous NaHCO3 to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by flash reverse column chromatography (CH3CN / H2O) to obtain the white solid P1 (2 mg), LC-MS: [M+H] + = 507.

[0283] 1 H NMR (400 MHz, DMSO-d6) δ 12.91 (s, 1H), 8.40 (s, 1H), 8.17 - 8.23 (d, J = 8.4 Hz, 1H), 8.02 - 8.08 (dd, J = 12.4, 4.6 Hz, 1H), 7.76 (s, 1H), 7.55 - 7.60 (d, J = 9.2 Hz, 1H), 4.26 - 4.38 (m, 2H), 4.23 - 4.26 (m, 1H), 3.82 (s, 3H), 1.83 - 1.88 (d, J = 14.0 Hz, 3H), 1.75 - 1.82 (d, J = 14.0 Hz, 3H), 0.94 - 0.98 (m, 2H), 0.90 - 0.92 (m, 2H).

[0284] Example 5: Synthesis of the target molecule P3

[0285]

[0286] Step 1: Under nitrogen protection, dissolve the compound M0-1 (100 mg, 0.18 mmol) and the raw material tri-n-butyltin propynide P3-1 (240 mg, 0.72 mmol) in 10 mL of 1,4-dioxane, add Pd(PPh3)4 (42 mg, 0.036 mmol), heat to 120 °C and react for 72 h, and monitor the reaction completion by LCMS. Add 30 mL of ice water to the system, extract with dichloromethane, dry over anhydrous sodium sulfate, concentrate, and separate by column chromatography (DCM / MeOH, 10 / 1) to obtain the pale yellow solid P3-2 (50 mg), yield: 49%. LC-MS: [M+H] + = 569. LC-MS: [M+Na] + = 591.

[0287] Step 2: Separate the compound P3-2 (60 mg) by SFC chiral column chromatography to obtain the compounds P3-3 (20 mg) and P3-4 (19 mg).

[0288] Resolution conditions: Prep-SFC (column: CHIRALPAK IE, 2 * 25 cm, 5 μm; mobile phase A: MtBE (0.5% 2M NH3-MeOH), mobile phase B: MeOH:DCM = 1:1; flow rate: 20 mL / min; retention time of P3-3 (min): 8.509; retention time of P3-4 (min): 11.694).

[0289] Step 3: Dissolve the intermediate P3-3 (20 mg, 0.04 mmol) from the previous step in 1 mL of 1,4-dioxane, add 1 mL of a 1,4-dioxane solution of hydrogen chloride (1 M), stir at room temperature for 1 hour, and detect the completion of the reaction by LCMS. Evaporate the solvent under reduced pressure, add 40 mL of saturated aqueous NaHCO3 to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by flash reverse column chromatography (CH3CN / H2O) to obtain white solid P3-a (9.1 mg), yield: 55%. LC-MS: [M + H] + = 469.

[0290] 1 H NMR (400 MHz, DMSO-d6) δ 12.46 (s, 1H), 8.26 (s, 1H), 8.16 (d, J = 8.2 Hz, 1H), 7.79 (d, J = 5.7 Hz, 1H), 7.73–7.66 (m, 2H), 4.16 (tt, J = 6.1, 3.0 Hz, 1H), 3.76 (d, J = 3.0 Hz, 5H), 2.18 (s, 3H), 1.89 (s, 2H), 0.92–0.79 (m, 4H).

[0291] Step 4: Refer to the synthetic route of compound P3-a to obtain compound P3-b (10.1 mg), yield: 61%. LC-MS: [M + H] + = 469.

[0292] 1 H NMR (400 MHz, DMSO-d6) δ 12.46 (s, 1H), 8.26 (s, 1H), 8.16 (d, J = 8.2 Hz, 1H), 7.79 (d, J = 5.6 Hz, 1H), 7.73–7.67 (m, 2H), 4.20–4.10 (m, 1H), 3.77 (s, 5H), 2.18 (s, 3H), 1.98 (d, J = 30.4 Hz, 2H), 0.94–0.87 (m, 2H), 0.80 (d, J = 3.6 Hz, 2H).

[0293] Refer to the synthetic route of compound P3-a or P3-b, and use similar raw materials or intermediates to synthesize the following target molecules.

[0294]

[0295]

[0296] Example 6: Synthesis of Target Molecules P7 - P13

[0297]

[0298] Step 1: Under nitrogen protection, dissolve intermediate a9 (258 mg, 0.6 mmol), intermediate a4 (392 mg, 0.98 mmol) and Na2CO3 (138 mg, 1.3 mmol) in a mixed solution of 5 mL of 1,4 - dioxane and water (v / v, 9 / 1). Add Pd(dtbpf)Cl2 (42 mg, 0.07 mmol), and heat the mixture to 80 °C and react for 2 hours. Cool to room temperature, add water to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by flash reverse column chromatography (CH3CN / H2O, 3 / 4) to obtain a pale yellow solid P7 - 1 (127 mg), yield: 33%. LC - MS: [M + H] + = 591.

[0299] Step 2: Under nitrogen protection, dissolve compound P7 - 1 (127 mg, 0.22 mmol) and raw material tri - n - butylpropynylstannane P3 - 1 (282 mg, 0.86 mmol) in 3 mL of 1,4 - dioxane. Add catalyst Pd(dppe)4 (25 mg, 0.02 mmol), and heat the mixture to 110 °C and react for 12 hours. Monitor the reaction completion by LCMS. Add 30 mL of ice - water to the system, extract with dichloromethane, dry over anhydrous sodium sulfate, concentrate, and separate by preparative chromatography (chromatographic column: GreenSep Naphthyl, 4.6 * 100 mm, 3 um; mobile phase B: MeOH(1% 2M NH3 - MeOH); flow rate: 4 mL / min;) to obtain a white solid P7 - 2 (15 mg), yield: 12%. LC - MS: [M + H] + = 595.

[0300] Step 3: Dissolve the intermediate P7-2 (15 mg, 0.03 mmol) from the previous step in 1 mL of 1,4-dioxane, add 1 mL of a 1,4-dioxane solution of hydrogen chloride (1 M), stir at room temperature for 1 hour, and detect the completion of the reaction by LCMS. Evaporate the solvent under reduced pressure, add 40 mL of saturated aqueous NaHCO3 to the system, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and separate by flash reverse column chromatography (CH3CN / H2O) to obtain the white solid hydrochloride P7 (8.9 mg), yield: 67%. LC-MS: [M+H] + = 495.

[0301] 1 H NMR (400 MHz, DMSO-d6) δ 12.90 (s, 1H), 8.50 (s, 3H), 8.32 (d, J = 1.9 Hz, 1H), 8.16 (d, J = 8.4 Hz, 1H), 7.89–7.75 (m, 2H), 7.44 (dd, J = 8.4, 1.6 Hz, 1H), 4.38 (s, 2H), 4.20 (s, 1H), 3.60–3.48 (m, 1H), 2.18 (s, 3H), 1.24 (s, 3H), 1.02 (d, J = 3.9 Hz, 2H), 0.92 (dd, J = 6.8, 3.7 Hz, 3H), 0.83 (t, J = 12.6 Hz, 2H).

[0302] Refer to the synthetic route of compound P7, and use similar raw materials or intermediates to synthesize the following target molecules.

[0303] * indicates a chiral center and has not been resolved.

[0304]

[0305] Example 7:

[0306] Intracellular Arginine Symmetric Dimethylation Inhibition Assay

[0307] Seed HCT116 wild-type or MTAP-deficient cells (HTC116-MTAP del) cultured in RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin on a 384-well microplate with a culture volume of 30 μL, and incubate overnight at 37 °C and 5% carbon dioxide. Add 60 nL of different concentrations of the compound to each well using ECHO, and incubate at 37 °C and 5% carbon dioxide for 96 hours.

[0308] Then, 50 μL of 4% paraformaldehyde solution was added to each well, and the cells were fixed by incubating at room temperature for 20 minutes. The solution in the wells was removed, and each well was washed 4 times with PBS solution containing 0.1% Tween 20 (PBST). 30 μL of methanol pre-cooled in an ice bath was added to each well and placed at -20 °C for incubation for 10 minutes; the methanol was removed and washed 4 times with PBST. 30 μL of Odyssey blocking solution containing 0.5% Tween 20 was added to each well, and the mixture was incubated with shaking at room temperature for 2 hours; the blocking solution was removed, and 30 μL of primary antibody (Symmetric Di-Methyl Arginine Motif [sdme-RG] MultiMab TM Rabbit mAb mix) diluted 1:500 in Odyssey blocking solution containing 0.5% Tween 20 was added to each well and incubated overnight at 4 °C; the primary antibody was removed, and each well was washed 4 times with PBST for 5 minutes each time; 30 μL of secondary antibody (goat anti-rabbit IRDye 800CW, 1:800) and nuclear stain (DRAQ5, 1:10,000) diluted in Odyssey blocking solution containing 0.5% Tween 20 were added to each well, and the mixture was incubated in the dark at room temperature for 2 hours. The secondary antibody was removed and washed 4 times with PBST.

[0309] The sdme-RG and DRAQ5 signals were scanned at 800 nm and 700 nm respectively using a Li-Cor Odyssey instrument and the signal values were recorded. The ratio of sdme-RG / DRAQ5 was used to calculate the inhibition percentage of arginine symmetric dimethylation (SDMA), and the IC 50 value was calculated using GraphPad Prism software.

[0310] Table 1: Inhibitory effect of compounds on arginine symmetric methylation in HTC116-MTAP del and wild-type colorectal cancer HCT-116 cell lines

[0311]

[0312] N.D. = Not tested

[0313] The above experimental results show that the excellent compounds of the present invention have an obvious effect of inhibiting arginine symmetric methylation in MTAP-deficient tumor cells by inhibiting the PRMT5-MTA complex, while having a weaker inhibitory effect on wild-type (less MTA).

[0314] The activities of some compounds are similar to or better than those of the control molecule M0-b.

[0315] MTA = Methylthioadenosine

[0316] Example 8:

[0317] HCT116 wild-type and MTAP-deficient cells were cultured in McCoy's 5A medium containing 10% FBS and 1% penicillin-streptomycin, placed in a 37°C, 5% CO2 incubator, and 40 μL of cell suspension was added to each well of a 384-well microplate. 40 nL of compounds at different concentrations were added to each well using an Echo, and the cells were cultured in a 37°C, 5% CO2 incubator for 7 days. 40 μL of CTG solution (Promega, Cat No. G7573) was added to each well, and the plate was incubated in the dark at 37°C, 5% CO2 for 30 minutes. The luminescence value was read using an Envision multimode microplate reader (Perkin Elmer, catalog number Envision 2104). The light signal is proportional to the amount of ATP in the system, and the content of ATP directly represents the number of viable cells in the system.

[0318] IC 50 Value calculation:

[0319] Y = lower plateau signal + (upper plateau signal - lower plateau signal) / (1 + 10^((LogIC 50 -X) × Hill slope))

[0320] X: Log value of compound concentration

[0321] Y: Inhibition rate (%)

[0322] Table 2: 2D anti-proliferation effects of compounds on HTC116-MTAP del and wild-type colorectal cancer HCT-116 cell lines

[0323] Compound <![CDATA[HCT116-MTAPdel / IC 50 / nM]]> <![CDATA[HCT116-MTAPwt / IC 50 / nM]]> P3-a 553 1332 P3-b 8 626 P4-a 460 1741 P4-b 31 764 P5-a 698 2399 P5-b 106 2657 P6-a 16 997 P6-b 810 9680 P7 474 5830 P8 965 5801 P9 2356 8205 P10 44 1896 P11 35 2336 P12 112 1954 P13 89 546

[0324] The above experimental results show that the excellent compounds of the present invention have obvious anti-proliferation effects on MTAP-deficient tumor cells by inhibiting the PRMT5-MTA complex.

[0325] Example 9:

[0326] Hepatic microsomal stability experiment of compounds. Specifically as follows:

[0327] The hepatic microsomal stability of the compounds of the present invention was studied. The test compounds were co-incubated with hepatic microsomes of different species with or without NADPH. The final concentration of the test compound in the test system was 1 μM, the final concentration of NADPH was 1 mM, and the final concentration of hepatic microsomes was 0.5 mg / mL. The concentration of the compound in the incubation supernatant at different time points within 60 minutes was detected and pharmacokinetic parameters (such as clearance Clint) were calculated.

[0328] The results indicate that the molecules of the present invention have good metabolic stability (especially in humans, with good metabolic stability).

[0329] Compound Human Clint / (mL / min / kg) P3-b 20.1 Control M0-b 22.9

[0330] Compared with the control molecule M0-b, some compounds are more stable in human metabolism.

[0331] Example 10:

[0332] Pharmacokinetics experiment of the compound in mice. The specific steps are as follows:

[0333] Using CD1 female mice as the test animals, oral / intravenous administration (oral dosage is 10 mg / kg, intravenous is 2 mg / kg). Oral solvent: 0.1% Tween 80 + 0.5% methylcellulose + saline; intravenous solvent: 5% DMSO + 95% "20% HP-β-CD saline".

[0334] Experimental protocol: Each group in the intravenous group has three mice. Collect plasma samples before dosing (0 h) and after dosing (0.083, 0.25, 0.5, 1, 2, 4, 8, 24 h); use LC / MS / MS method to measure the blood concentration in the plasma of mice after intravenous administration respectively, and the collected data is calculated by AB Sciex QTRAP 6500 software. The experimental results are as follows:

[0335]

[0336] The above experimental results show that, compared with the control molecule M0-b, the compounds of the present invention in mice are consistent with in vitro microsomes, with a lower in vivo clearance rate and a higher exposure.

[0337] Example 11:

[0338] Efficacy experiment in BALB / c nude mice. The specific steps are as follows:

[0339] Cultivate HCT116 (MTAPdel) tumor cells and inoculate these tumor cells into female BALB / c nude mice at 6 - 8 weeks old (with a body weight of about 20 g), and all mice are inoculated subcutaneously. The mice are cultured in a SPF - level experimental environment, and all mice can freely access a commercially certified standard diet. When the average tumor volume of the mice grows to about 160 mm3, the test compound is started to be administered orally daily. The dosing regimens are as follows: the vehicle for the blank group is Saline containing 0.1% Tween 80 and 0.5% methyl cellulose. The dosing regimen for the P3 - b group is 100 mg / kg once a day, and the dosing regimen for the positive control drug GSK3326595 is 100 mg / kg twice a day. The tumor volume is measured three times a week using a two - dimensional caliper, and the animals are weighed every day. After 19 days of continuous dosing, the inhibition rate (TGI / 100%) is calculated based on the final tumor volume. The volume calculation formula is: V = 1 / 2a*b 2 , where a represents the long diameter of the tumor and b represents the short diameter of the tumor.

[0340] Test drug Dosing dose <![CDATA[Tumor volume (mm 3 )]]> TGI Blank group 0 1087 0％ P3-b 100 mg / kg, QD 257 76％ GSK3326595 100 mg / kg, BID 378 65％

[0341] The results indicate that the molecule of the present invention has good in - vivo efficacy, which is superior to the drug GSK3326595 that has been in clinical development.

[0342] Structure of GSK3326595:

Claims

1. A compound, or a tautomer or pharmaceutically acceptable salt thereof, wherein the compound is selected from:

2. A pharmaceutical composition comprising the compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition of claim 2, wherein, The composition further contains other therapeutic agents.

4. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment and / or prevention of diseases mediated by PRMT5 methyltransferase.

5. The use of claim 4, wherein the disease mediated by PRMT5 methyltransferase is cancer, and the cancer is selected from: acoustic neuroma, adenocarcinoma, adrenal cancer, anal cancer, angiosarcoma, appendiceal cancer, benign monoclonal gammopathy, cholangiocarcinoma, bladder cancer, brain cancer, bronchial cancer, carcinoid tumor, cervical cancer, choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer, epithelial cancer, ependymoma, endothelial sarcoma, endometrial cancer, esophageal cancer, Ewing's sarcoma, eye cancer, eosinophilia, gallbladder cancer, gastric cancer, gastrointestinal stromal tumor (GIST), head and neck cancer, oral cancer, laryngeal cancer, hematopoietic system cancer, multiple myeloma (MM), heavy chain disease, hemangioblastoma, inflammatory myofibroblastic tumor, immunocytic amyloidosis, kidney cancer, liver cancer, lung cancer, lung adenocarcinoma, leiomyosarcoma (LMS), mastocytosis, myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disease (MPD), neuroblastoma, neurofibroma, neuroendocrine cancer, osteosarcoma, ovarian cancer, papillary adenocarcinoma or penile cancer.

6. Use according to claim 5, wherein, The angiosarcoma is selected from lymphangiosarcoma, lymphatic endothelial sarcoma or hemangioma.

7. Use according to claim 5, wherein, The brain cancer is selected from meningioma or glioma.

8. Use according to claim 7, wherein, The glioma is selected from astrocytoma, oligodendroglioma or medulloblastoma.

9. Use according to claim 5, wherein, The cervical cancer is cervical adenocarcinoma.

10. Use according to claim 5, wherein, The colorectal cancer is selected from colon cancer, rectal cancer or colorectal adenocarcinoma.

11. Use according to claim 5, wherein, The endothelial sarcoma is selected from Kaposi's sarcoma or multiple idiopathic hemorrhagic sarcoma.

12. Use according to claim 5, wherein, The endometrial cancer is selected from uterine cancer or uterine sarcoma.

13. Use according to claim 5, wherein, The esophageal cancer is selected from esophageal adenocarcinoma or Barrett's adenocarcinoma.

14. Use according to claim 5, wherein, The eye cancer is selected from intraocular melanoma or retinoblastoma.

15. Use according to claim 5, wherein, The gastric cancer is gastric adenocarcinoma.

16. Use according to claim 5, wherein, The head and neck cancer is head and neck squamous cell carcinoma.

17. Use according to claim 5, wherein, The oral cancer is oral squamous cell carcinoma.

18. Use according to claim 5, wherein, The laryngeal cancer is selected from pharyngeal cancer, nasopharyngeal cancer or oropharyngeal cancer. Use according to claim 5, wherein, The hematopoietic system cancer is selected from leukemia and T-cell non-Hodgkin lymphoma. Use according to claim 5, wherein, The hematopoietic system cancers are selected from acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), marginal zone B-cell lymphoma, primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma, precursor T lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma, angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma or anaplastic large cell lymphoma. Use according to claim 5, wherein, The hematopoietic system cancers are selected from B-cell ALL, T-cell ALL, B-cell AML, T-cell AML, B-cell CML, T-cell CML, B-cell CLL, T-cell CLL, mucosa-associated lymphoid tissue (MALT) lymphoma, nodal marginal zone B-cell lymphoma or splenic marginal zone B-cell lymphoma.

22. Use according to claim 20, wherein, The peripheral T-cell lymphoma is selected from cutaneous T-cell lymphoma, angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma or anaplastic large cell lymphoma.

23. Use according to claim 22, wherein, The cutaneous T-cell lymphoma is selected from mycosis fungoides or Sézary syndrome.

24. Use according to claim 5, wherein, The heavy chain diseases are selected from alpha chain disease, gamma chain disease or mu chain disease. Use according to claim 5, wherein, The renal cancers are selected from nephroblastoma or renal cell carcinoma.

26. Use according to claim 5, wherein, The liver cancers are selected from hepatocellular carcinoma or malignant hepatocellular carcinoma. Use according to claim 5, wherein, The lung cancers are selected from bronchial carcinoma, small cell lung cancer (SCLC) or non-small cell lung cancer (NSCLC). Use according to claim 5, wherein, The mastocytosis is systemic mastocytosis. Use according to claim 5, wherein, The myeloproliferative diseases (MPD) are selected from polycythemia vera (PV), essential thrombocythemia (ET), idiopathic myelofibrosis with myeloid metaplasia (AMM), chronic idiopathic myelofibrosis or hypereosinophilic syndrome (HES). Use according to claim 5, wherein, The neurofibromas are selected from neurofibromatosis type 1 or type 2 or schwannomatosis.

31. Use according to claim 5, wherein, The neuroendocrine cancers are selected from gastroenteropancreatic neuroendocrine tumors (GEP-NET) or carcinoid tumors. Use according to claim 5, wherein, The ovarian cancers are selected from cystadenocarcinoma, ovarian embryonal carcinoma or ovarian adenocarcinoma.

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