Substituted fused ring compounds and pharmaceutical compositions, methods of manufacture and uses thereof

Abstract

The present invention provides a substituted fused ring compound represented by formula (I) and its pharmaceutical composition, preparation method and use, which has good LRRK2 kinase regulating / inhibiting activity and can be used for treating symptoms and diseases associated with LRRK2 kinase activity, and for producing drugs for such symptoms and diseases.

Description

Detailed Description of the Invention

[0001] This application claims priority to two prior Chinese patent applications, namely, an earlier application filed by the applicant with the State Intellectual Property Office on February 25, 2022, bearing application number 202210178818.7 and entitled "Substituted fused ring compounds and pharmaceutical compositions, preparation methods and use thereof," and an earlier application filed with the State Intellectual Property Office on April 28, 2022, bearing application number 202210470959.6 and entitled "Substituted fused ring compounds and pharmaceutical compositions, preparation methods and use thereof." Both of the above two prior patent applications are incorporated herein by reference in their entireties.

[0002] [Technical field] The present disclosure relates to the field of LRRK2 kinase drugs, specifically to substituted fused ring compounds and their pharmaceutical compositions, methods of manufacture and uses.

[0003] [Background technology] Parkinson's disease (PD) is one of the common neurodegenerative disorders, affecting 1-2% of the elderly population. Genome-wide association studies (GWAS) have linked 28 genetic risk variants at 24 sites to non-familial PD, among which LRRK2 (Park8) mutations were also found in the inherited form, confirming a common molecular pathway-driven pathogenesis for familial and non-familial PD, and are the most common cause of associated disease (Simon-Sanchez J, Schulte C, Bras JM, Sharma M, Gibbs JR, et al. Genome-wide association study reveals genetic risk underlying Parkinson's disease. Nat Genet, 2009, 41, 1308-1312.). PD pathogenic LRRK2 mutations map mainly to the kinase (G2019S, I2020T) and ROC-COR domains (R1441C / G / H, Y1699C), implying that the activity of these enzymes is crucial for the pathology. The frequency of pathogenic mutations is very rare, accounting for approximately 2% of all cases, but in some ethnic groups, the most common mutation G2019S is found in up to 40% of patients and activates the kinase 2-3-fold (West AB, Moore DJ, Biskup S, Bugayenko A, Smith WW, et al. From The Cover: Parkinson's disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity. Proceedings of the National Academy of Sciences, 2005, 102, 16842-16847.).In addition to pathogenic mutations, common genetic variants in LRRK2 are risk factors for sporadic PD (Tan, EK Identification of a common genetic risk variant(LRRK2 Gly2385Arg)in Parkinson's disease. Ann Acad Med Singapore. 2006, 35, 840-842.).

[0004] In 2004, LRRK2 was identified as the gene responsible for PD inheritance, and was found to be correlated with the PARK8 locus and composed of 51 exons, producing one large (268 kDa) protein. Since then, many LRRK2 primary structure variants have been identified, including dominant mutations distant from familial PD that also appear in sporadic PD, and polymorphisms in the LRRK2 locus, which increase the lifetime risk of developing sporadic PD.

[0005] LRRK2 is a multidomain protein with two enzymatic functions in its core. The GTPase domain contains the Ras of complex protein (ROC), which terminates in a spacer domain at the C-terminus of the Roc domain (COR), followed by a kinase domain belonging to a serine / threonine kinase. The enzymatic core is surrounded by protein-protein interaction domains, including Armadillo, ankyrin, and a leucine-rich repeat (LRR) domain at the N-terminus of LRRK2. The C-terminus of LRRK2 contains a WD40 domain, a region required for protein folding and capable of regulating LRRK2 function and kinase activity (Rudenko IN, Kaganovich A, Hauser DN, Beylina A, Chia R, et al. The G2385R variant of leucine-rich repeat kinase 2 associated with Parkinson's disease is a partial loss-of-function mutation. Biochemical Journal, 2012, 446, 99-111.). Interestingly, the new description of dominant pathogenic mutations occurring within the enzymatic core of LRRK2 reveals that modification of LRRK2 activity has a profound effect on the development and progression of PD.

[0006] To date, nearly 40 single amino acid substitution mutations in LRRK2 have been associated with autosomal dominant PD. The most common LRRK2 mutation type in Europe involves the substitution of the amino acid Gly2019 to Ser residue, accounting for approximately 6% of familial PD cases and 3% of sporadic PD cases. Gly2019 is a conserved DYG-Mg 2+It is located within the binding motif and in subdomain-VII of the kinase domain. Recent reports reveal that such mutations enhance LRRK2 autophosphorylation and increase its ability to phosphorylate myelin basic protein by 2-3 fold. When G2019S-LRRK2 is overexpressed in cell lines and primary neuronal cultures, both in the presence and absence of oxidative stress, cytotoxicity and inclusion formation are observed. These results, together with the genetic inactivation of LRRK2 kinase activity showing a protective effect against such toxic phenotypes, reveal that alterations in LRRK2 kinase activity may be relevant to the neurotoxicity and pathogenic mechanisms of LRRK2-PD.

[0007] Studies have found that induced pluripotent stem cells (iPSCs) from LRRK2 G2019S Parkinson's disease patients exhibit neurite outgrowth defects and increased sensitivity to rotenone, which can be ameliorated by genetic correction of the G2019S mutation or by treating the cells with small molecule inhibitors of LRRK2 kinase activity (Reinhardt P, Schmid B, Burbulla Lena F, Schoendorf David C, Wagner L, et al. Genetic Correction of a LRRK2 Mutation in Human iPSCs Links Parkinsonian Neurodegeneration to ERK-Dependent Changes in Gene Expression. Cell Stem Cell, 2013, 12, 354-367.). The increased mitochondrial damage correlated with the LRRK2 G2019S mutation in iPSCs is also blocked by genetic correction of the G2019S mutation.

[0008] Summary of the Invention In order to improve the above technical problems, the present disclosure provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester, or a prodrug compound thereof,

[0009] [ka]

[0010] Among them, W is selected from N or O; R 1 is unsubstituted or contains one, two or more R a C replaced with 1-10 Alkyl group, C 3-10 a cycloalkyl group, or a 3- to 10-membered heterocyclyl group; Each R a are identical or different and independently represent a halogen, a hydroxyl group, C 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, and a 3- to 10-membered heterocyclyloxy group; Or, alternatively, two R a are unsubstituted or optionally substituted with one, two or more R c C replaced with 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 forming a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, or a 3- to 10-membered heterocyclyloxy group; R 2 is absent or is H or C 1-10 alkyl groups, in which when W is selected from O, R 2 does not exist and W is chosen from N, then R 2 is H or C 1-10 selected from alkyl groups, Or, alternatively, R 1 , R 2 is unsubstituted or optionally one, two or more R b forming a 3- to 10-membered heterocyclyl group substituted with Each R b are identical or different and independently represent a halogen, a hydroxyl group, C 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, and a 3- to 10-membered heterocyclyloxy group; Or, alternatively, two R b are unsubstituted or optionally substituted with one, two or more R c C replaced with 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 forming a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, or a 3- to 10-membered heterocyclyloxy group; Each R c are identical or different and independently represent a halogen, a hydroxyl group, C 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, and a 3- to 10-membered heterocyclyloxy group; X is one, two or more R 3 substituted 5-10 membered heteroaryl groups, Each R 3 are identical or different and are independently unsubstituted or substituted with one, two or more R 4 C replaced with 1-10 Alkyl group, C 3-10 selected from a cycloalkyl group, a 3- to 10-membered heterocyclyl group, and a 5- to 10-membered heteroaryl group; Or, alternatively, two R 3are unsubstituted or optionally substituted with one, two or more R 4 C replaced with 3-10 Forming a cycloalkyl group or a 3- to 10-membered heterocyclyl group; Each R 4 are identical or different and independently represent a halogen, a hydroxyl group, a cyano group, no substitution or one, two or more R d C replaced with 1-10 Alkyl group, C 1-10 Alkylene group, C 3-10 A cycloalkyl group, a 3- to 10-membered heterocyclyl group, and a 5- to 10-membered heteroaryl group, among which R 4 C 1-6 When selected from alkylene groups, R 4 The carbon atom in R 3 means to form a double bond with the carbon atom linked to Each R d are identical or different and each independently represents a halogen, a hydroxyl group, a cyano group, C 1-6 Alkyl group, C 1-6 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 It is selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, a 3- to 10-membered heterocyclyloxy group, a 5- to 10-membered heteroaryl group, and a 5- to 10-membered heteroaryloxy group.

[0011] According to an embodiment of the present disclosure, R 1 is unsubstituted or contains one, two or more R a C replaced with 1-8 Alkyl group, C 3-8 a cycloalkyl group, and a 3- to 8-membered heterocyclyl group; For example, R 1 is unsubstituted or contains one, two or more R a C replaced with 1-6 Alkyl group, C 3-7 a cycloalkyl group, and a 3- to 7-membered heterocyclyl group; According to an embodiment of the present disclosure, each R aare identical or different and independently represent F, Cl, Br, C 1-6 selected from alkyl groups, For example, R 1 teeth,

[0012] [ka]

[0013] -CH2CF3, -CH(CH3)CF3.

[0014] According to an embodiment of the present disclosure, R 1 , R 2 is unsubstituted or optionally one, two or more R b Replaced with

[0015] [ka]

[0016] Form.

[0017] According to an embodiment of the present disclosure, X is one, two or more R 3 substituted 5-6 membered heteroaryl groups, For example, X can be one, two or more R 3 Replaced with

[0018] [ka]

[0019] Selected from For example, X is

[0020] [ka]

[0021] Selected from.

[0022] According to an embodiment of the present disclosure, R 3 is unsubstituted or contains one, two or more R 4 C replaced with 1-6 Alkyl group, C 3-8 selected from a cycloalkyl group, a 3- to 8-membered heterocyclyl group, and a 5- to 6-membered heteroaryl group; For example, R 3 is unsubstituted or contains one, two or more R 4 methyl, ethyl, propyl, isopropyl, cyclopropane, cyclobutane substituted with

[0023] [ka]

[0024] Selected from Each R 4 are identical or different and independently represent fluorine, chlorine, bromine, a hydroxyl group, a cyano group, unsubstituted or one, two or more R d C replaced with 1-6 Alkyl group, C 1-6 Alkylene group, C 3-8 a cycloalkyl group, and a 3- to 8-membered heterocyclyl group; For example, R 3 is an isopropyl group,

[0025] [ka]

[0026] Selected from.

[0027] According to a preferred embodiment of the present disclosure, the compound represented by formula (I) is preferably a compound represented by the following formula (II):

[0028] [ka]

[0029] Among them, R 5 is C substituted with one, two or more halogens 1-6 Alkyl groups, preferably C substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 halogens 1-6 alkyl groups, for example methyl or ethyl groups substituted by 1, 2, 3, 4 or 5 fluorine or chlorine atoms, for example trifluoromethyl groups, R 6 is C 1-6 alkyl groups, such as methyl, ethyl, propyl or isopropyl groups; R 7 , R 8 , R 9 , R 10 are homologous or different and independently represent H or C 1-6 an alkyl group, for example selected from H, methyl, ethyl, propyl or isopropyl; R 11 , R 12 are homologous or different and independently represent H or C 1-6 Alkyl groups, for example, selected from H, methyl, ethyl, propyl or isopropyl groups.

[0030] According to a preferred embodiment of the present disclosure, the compound represented by formula (I) is preferably a compound represented by the following formula (III):

[0031] [ka]

[0032] Among them, R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 has the definition set forth above.

[0033] According to an exemplary embodiment of the present disclosure, the compound represented by formula (I) is the following compound:

[0034] [ka] JPEG2025506822000011.jpg235169JPEG2025506822000012.jpg80169

[0035] may be selected from.

[0036] The present disclosure further provides a process for preparing a compound of formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester, or a prodrug compound thereof, comprising the reaction:

[0037] [ka]

[0038] Among them, R 1 , R 2 , W and X have the definitions described above; P 1 is selected from amino protecting groups.

[0039] Preferably, the compound of formula (M2) is a protecting group P 1 is reacted under conditions that eliminate the aryl group to obtain a compound represented by formula (I).

[0040] According to an embodiment of the manufacturing method of the present disclosure, the above P 1is selected from amino protecting groups known to those skilled in the art. For example, suitable amino protecting groups may be selected from alkyloxycarbonyl groups (e.g., tert-butoxycarbonyl group, etc.), cycloalkylalkoxycarbonyl groups (e.g., cyclohexylmethoxycarbonyl group), substituted alkyloxycarbonyl groups (e.g., trichloroethoxycarbonyl group, etc.), substituted arylalkyloxycarbonyl groups (e.g., p-nitrobenzyloxycarbonyl group), arylalkyl groups (e.g., triphenylmethyl group or diphenylmethyl group), heteroarylalkyl groups (e.g., pyridylalkyl groups, e.g., 2-pyridylmethyl group, 4-pyridylmethyl group), alkoxyarylalkyl groups (e.g., alkoxyphenylalkyl groups, e.g., p-methoxybenzyl group, abbreviated as "PMB"), and the like.

[0041] According to an embodiment of the process of the present disclosure, there is further provided a process for preparing a compound of formula (M2), comprising the reaction:

[0042] [ka]

[0043] Among them, R 1 , R 2 , W, X and P 1 has the definition set forth above, L 1 and L 2 are the same or different and are selected from leaving groups which enable the coupling reaction independently of each other.

[0044] According to an embodiment of the present disclosure, the preparation of the compound of formula (M2) can be carried out by coupling reactions known in the art, for example, Stille coupling reaction or Suzuki coupling reaction.

[0045] As the Stille coupling reaction or Suzuki coupling reaction known to those skilled in the art, L 1 and L 2is known to those skilled in the art. For example, L 1 may be selected from halogens such as iodine, L 2 may be selected from an alkyl tin group or a borate ester group.

[0046] The present disclosure further provides a pharmaceutical composition comprising at least one of the compounds of formula (I), their racemates, stereoisomers, tautomers, isotopically labeled compounds, N-oxides, hydrates, solvates, crystalline polymorphs, metabolites, pharma- ceutically acceptable salts, pharma- ceutically acceptable esters, or prodrug compounds thereof.

[0047] The present disclosure further provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds of formula (I), its racemates, stereoisomers, tautomers, isotopically labeled compounds, N-oxides, hydrates, solvates, crystalline polymorphs, metabolites, pharma- ceutically acceptable salts, pharma-ceutically acceptable esters, or prodrug compounds thereof.

[0048] According to an embodiment of the present disclosure, the pharmaceutical composition further comprises one or more pharma- ceutically acceptable adjuvants.

[0049] According to embodiments of the present disclosure, the pharmaceutical composition may further comprise one or more additional therapeutic agents.

[0050] According to embodiments of the present disclosure, a compound of formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester or prodrug compound thereof, or the pharmaceutical composition described above, can be used to modulate the activity of LRRK kinase, particularly LRRK2 kinase.

[0051] According to an embodiment of the present disclosure, the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester or prodrug compound thereof, or said pharmaceutical composition can be used for the prophylaxis and / or treatment of a disease or condition, including, but not limited to, a neurodegenerative disease, a proliferative disease, a disease associated with a protein kinase, a lysosomal disease, a tau disease, and a dopaminergic disease. The present invention may include one, two or more selected from diseases caused by decreased alpha-synuclein levels, such as cancer, Parkinson's disease (PD) associated with GBA mutations, other alpha-synucleinopathies, tauopathies, Alzheimer's disease, Gaucher disease, Niemann-Pick disease type C (NPC), argyrophilic grain disease, Pick's disease, corticobasal degeneration, progressive supranuclear palsy, hereditary frontotemporal dementia and Parkinson's disease linked to chromosome 17 (FTDP-17), and withdrawal / relapse associated with drug addiction.

[0052] The disclosure further provides methods for modulating the activity of LRRK kinase, particularly LRRK2 kinase, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one of a compound of formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester, or prodrug compound thereof.

[0053] According to an embodiment of the present disclosure, the LRRK kinase is preferably LRRK2 kinase or a mutant or isoform thereof, or a combination thereof.

[0054] The disclosure further provides at least one of a compound of formula (I), a racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester or prodrug compound thereof, or a pharmaceutical composition thereof, for use in treating a disease or condition associated with LRRK kinase, particularly LRRK2 kinase activity.

[0055] According to an embodiment of the present disclosure, the LRRK kinase is preferably LRRK2 kinase or a mutant or isoform thereof, or a combination thereof.

[0056] The above-mentioned diseases or conditions associated with LRRK kinase, particularly LRRK2 kinase activity, include one, two or more selected from neurodegenerative diseases, proliferative diseases, protein kinase-associated diseases, lysosomal diseases, tauopathies and diseases caused by reduced dopamine levels, such as cancer, Parkinson's disease (PD) associated with GBA mutations, other α-synucleinopathies, tauopathies, Alzheimer's disease, Gaucher disease, Niemann-Pick disease type C (NPC), argyrophilic grain disease, Pick's disease, corticobasal degeneration, progressive supranuclear palsy, hereditary frontotemporal dementia and Parkinson's disease linked to chromosome 17 (FTDP-17), withdrawal / relapse associated with drug addiction, and the like.

[0057] The present disclosure relates to a method for preventing and / or treating a disease or condition comprising administering to a patient at least one of a compound of formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester, or prodrug compound thereof, wherein the disease or condition is selected from the group consisting of neurodegenerative diseases, proliferative diseases, protein kinase-associated diseases, lysosomal diseases, tau diseases, and dopaminergic diseases. The present invention further provides a method for treating a disease caused by decreased pacin levels, which may include one, two or more selected from the group consisting of cancer, Parkinson's disease (PD) associated with GBA mutations, other α-synucleinopathies, tauopathies, Alzheimer's disease, Gaucher disease, Niemann-Pick disease type C (NPC), argyrophilic grain disease, Pick's disease, corticobasal degeneration, progressive supranuclear palsy, hereditary frontotemporal dementia and Parkinson's disease linked to chromosome 17 (FTDP-17), withdrawal / relapse associated with drug addiction, and the like.

[0058] In some embodiments, the patient with the disease or condition is a human.

[0059] The disclosure further provides a method for treating or preventing a disease or condition associated with LRRK kinase, particularly LRRK2 kinase activity, comprising administering to a patient at least one of a compound of formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester, or prodrug compound thereof.

[0060] The present disclosure further provides the use of at least one of the compounds of formula (I), its racemates, stereoisomers, tautomers, isotopically labeled compounds, N-oxides, hydrates, solvates, crystalline polymorphs, metabolites, pharma- ceutically acceptable salts, pharma- ceutically acceptable esters or prodrug compounds thereof in the manufacture of a medicament.

[0061] The above-mentioned drugs can be used for the prevention and / or treatment of diseases or conditions associated with LRRK kinase, particularly LRRK2 kinase activity.

[0062] Alternatively, the drug may be used for the prevention and / or treatment of a disease or condition, which may include one, two or more selected from neurodegenerative diseases, proliferative diseases, diseases associated with protein kinases, lysosomal diseases, tauopathies and diseases caused by decreased dopamine levels, such as cancer, Parkinson's disease (PD) associated with GBA mutations, other α-synucleinopathies, tauopathies, Alzheimer's disease, Gaucher disease, Niemann-Pick disease type C (NPC), argyrophilic grain disease, Pick's disease, corticobasal degeneration, progressive supranuclear palsy, hereditary frontotemporal dementia and Parkinson's disease associated with chromosome 17 (FTDP-17), withdrawal / relapse associated with drug addiction, and the like.

[0063] When used as a drug, the compounds of the present disclosure can be administered in the form of pharmaceutical compositions. These compositions can be prepared according to methods well known in the pharmaceutical art and can be administered via various routes depending on whether local or systemic treatment is required and the area to be treated. Administration can be topical (e.g., transdermal, to the skin, eyes, and mucous membranes, including intranasal, vaginal, and rectal), pulmonary (e.g., by inhaling or blowing powders or aerosols, including by nebulizer; intratracheal, intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion, or intracranial, such as intrathecal or intraventricular administration. They can be administered parenterally in a single bolus form or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, powder formulations, and powders. Conventional pharmaceutical carriers, water, powder or oily bases, thickeners and the like may be necessary or required.

[0064] In preparing the compositions of the present disclosure, the active ingredient is generally mixed with an excipient, diluted by the excipient, or placed in a carrier, such as, for example, a capsule, sachet, paper, or other container form. When used as a diluent, the excipient may be a solid, semi-solid, or liquid substance used as a solvent, carrier, or vehicle for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, topical applications, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or liquid solvents), ointments containing up to 10% by weight of the active compound, soft or hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaged powders.

[0065] Some examples of suitable excipients include lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulation may further include lubricants such as talc powder, sodium stearate, magnesium stearate, sodium oleate, sodium benzoate, sodium acetate, sodium chloride, and mineral oil, wetting agents, emulsifying and suspending agents, preservatives such as methyl benzoate and hydroxypropyl benzoate, sweeteners, and flavoring agents. The compositions of the present disclosure can be prepared in a manner known in the art so as to provide immediate, sustained, or delayed release of the active ingredient after administration to a patient.

[0066] The pharmaceutical compositions may be prepared in unit dosage form, each unit dosage containing from about 1 to 1000 mg, for example, about 5 to 1000 mg, more typically about 5 to 500 mg, for example 5 to 100 mg, 100 to 500 mg or 50 to 200 mg of the active ingredient. Illustratively, each dose may contain 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg or 300 mg of active ingredient. The term "unit dosage form" refers to physically discrete single-dose units suitable for use in human patients and other mammals, each unit containing a predetermined quantity of active material(s) mixed with a suitable pharmaceutical excipient(s) calculated to produce the desired therapeutic effect.

[0067] The effective dosage range of the active compound may be very large and is usually administered in a pharmacologic effective amount. However, it will be understood that the amount of the compound actually administered will usually be determined by the physician depending on the relevant circumstances, including the disease being treated, the route of administration selected, the actual compound being administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc.

[0068] The therapeutic dose of the compounds of the present disclosure can be determined, for example, by the specific use of the treatment, the method of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compounds of the present disclosure in a pharmaceutical composition may vary and depends on a variety of factors, including dose, chemical properties (e.g., hydrophobicity), and route of administration. For example, at least one of the compounds of the present disclosure, its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma-ceutically acceptable salt, pharma-ceutically acceptable ester, or prodrug compound thereof, can be provided for parenteral administration by an aqueous physiological buffer solution containing about 0.1-10% (w / v) of the compound. Some typical dose ranges are about 1 μg / kg to about 1 g / kg body weight / day. In some embodiments, the dose range is from about 0.01 mg / kg to about 100 mg / kg body weight per day, e.g., 0.1 mg / kg body weight per day, 0.2 mg / kg body weight per day, 0.3 mg / kg body weight per day, 0.4 mg / kg body weight per day, 0.5 mg / kg body weight per day, 0.6 mg / kg body weight per day, 0.7 mg / kg body weight per day, 0.8 mg / kg body weight per day, 0.9 mg / kg body weight per day, 1 mg / kg body weight per day, 5 mg / kg body weight per day, 10 mg / kg body weight per day, 15 mg / kg body weight per day, 20 mg / kg body weight per day, 25 mg / kg body weight per day, 30 mg / kg body weight per day, 35 mg / kg body weight per day, 40 mg / kg body weight per day, 45 mg / kg body weight per day, or 50 mg / kg body weight per day. By way of illustration, if the subject is a human, the dose administered to the patient may be 57 mg / 70 kg to 171 mg / 70 kg. The dose will likely depend on such variables as, for example, the type and extent of the disease or condition, the general health of the particular patient, the relative biological potency of the compound selected, the excipient formulation and its route of administration. Effective doses may be obtained by extrapolation from dose-response curves derived from in vitro or animal model test systems.

[0069] For the preparation of solid compositions such as tablets, the primary active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition that is a homogenous mixture containing a compound of the present disclosure. When these preformulation compositions are referred to as homogenous, it is meant that the active ingredient is generally uniformly distributed throughout the composition such that the composition may be readily divided into equally effective unit dosage forms such as tablets, pills, and capsules. The solid preformulation is then divided into unit dosage forms of the type described above, containing, for example, about 0.1-1000 mg of the active ingredient of the present disclosure, with each unit dosage form containing about 1-1000 mg, for example about 5-1000 mg, more typically about 5-500 mg, for example 5-100 mg, 100-500 mg, or 50-200 mg of active ingredient. Illustratively, each dose may contain 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg or 300 mg of active ingredient.

[0070] The tablets or pills of the present disclosure can be coated or compounded to obtain a dosage form that provides the advantage of sustained action. For example, the tablet or pill can include an inner dosage composition and an outer dosage composition, the latter being a coating form of the former. The two compositions can be separated by an enteric layer to prevent disintegration in the stomach, so that the inner composition passes completely through the duodenum or is released late. Such enteric layers or coatings can use a variety of materials, including a variety of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0071] Liquid forms for oral or injectable administration into which the compounds and compositions of the present disclosure may be incorporated include aqueous solutions, suitably flavored syrups, water or oil suspensions, emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, elixirs and similar pharmaceutical vehicles.

[0072] Compositions for inhalation or blowing include solutions and suspensions in pharma- ceutically acceptable water or organic solvents or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharma- ceutically acceptable excipients as described above. In some embodiments, the compositions are administered via the oral or nasal respiratory route to achieve a local or systemic effect. The compositions can be atomized by using an inert gas. The atomized solution may be directly inhaled by the atomizing device, or the atomizing device may be connected to a face mask tent or intermittent positive pressure respirator. The solution, suspension, or powder compositions may be administered orally or nasally by a device that delivers the formulation in an appropriate manner.

[0073] The amount of the compound, its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester or prodrug compound thereof, or pharmaceutical composition administered to a patient is not fixed and depends on the drug administered, the purpose of administration, such as prevention or treatment, the condition of the patient, the method of administration, and the like. In therapeutic applications, a sufficient amount of the composition to cure or at least partially inhibit the symptoms of the disease and its complications can be administered to a patient suffering from the disease. The effective dose should depend on the condition of the disease being treated and the judgment of the attending clinician, which depends on factors such as, for example, the severity of the disease, the age, weight, and general condition of the patient. Some typical dose ranges are from about 1 μg / kg to about 1 g / kg body weight / day. In some embodiments, the dose range is from about 0.01 mg / kg to about 100 mg / kg body weight per day, e.g., 0.1 mg / kg body weight per day, 0.2 mg / kg body weight per day, 0.3 mg / kg body weight per day, 0.4 mg / kg body weight per day, 0.5 mg / kg body weight per day, 0.6 mg / kg body weight per day, 0.7 mg / kg body weight per day, 0.8 mg / kg body weight per day, 0.9 mg / kg body weight per day, 1 mg / kg body weight per day, 5 mg / kg body weight per day, 10 mg / kg body weight per day, 15 mg / kg body weight per day, 20 mg / kg body weight per day, 25 mg / kg body weight per day, 30 mg / kg body weight per day, 35 mg / kg body weight per day, 40 mg / kg body weight per day, 45 mg / kg body weight per day, or 50 mg / kg body weight per day. By way of example, if the subject is a human, the dosage administered to the patient may be between 57 mg / 70 kg and 171 mg / 70 kg.

[0074] The compositions administered to a patient may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or sterile filtration. The aqueous solutions may be packaged as is or lyophilized and the lyophilized formulation may be mixed with a sterile aqueous carrier prior to administration. The pH of the compound formulation is generally 3-11, more preferably 5-9, and most preferably 7-8. It is understood that pharmaceutical salts may be formed using some of the above-mentioned excipients, carriers, or stabilizers.

[0075] The present disclosure provides the use of at least one of the compounds according to formula (I), its racemates, stereoisomers, tautomers, isotopically labeled compounds, N-oxides, hydrates, solvates, crystalline polymorphs, metabolites, pharma- ceutically acceptable salts, pharma- ceutically acceptable esters or prodrug compounds thereof in an analytical test, said analytical test being used to identify compounds capable of inhibiting one or more kinases, said kinase being preferably LRRK, more preferably LRRK2.

[0076] Preferably, the analytical assay is a competitive binding assay.

[0077] More preferably, the competitive binding assay involves contacting a compound of the present disclosure with a kinase, and detecting any change in the interaction between a compound of the present disclosure and the kinase.

[0078] Another aspect of the disclosure provides a method for detecting binding of a compound of the disclosure to a kinase, the method comprising: (i) contacting at least one of a compound of the present disclosure, a racemate, a stereoisomer, a tautomer, an isotopically labeled compound, an N-oxide, a hydrate, a solvate, a crystalline polymorph, a metabolite, a pharma- ceutically acceptable salt, a pharma-ceutically acceptable ester, or a prodrug compound thereof, with a kinase in the presence of the kinase and a known substrate; (ii) detecting any change in the interaction between the kinase and the known substrate.

[0079] Beneficial effects The compounds provided by the present disclosure have good LRRK2 kinase regulating / inhibiting effects, and can be used to treat symptoms and diseases associated with LRRK2 kinase activity, and to manufacture drugs for such symptoms and diseases. The compounds of the present disclosure have a simple preparation method and good application prospects.

[0080] Definitions and Explanations of Terms Unless otherwise specified, the definitions of groups and terms described in the specification and claims of this application, including their illustrative definitions, exemplary definitions, preferred definitions, definitions described in the tables, definitions of specific compounds in the examples, etc., can be arbitrarily combined or combined with each other. Such combinations and combined group definitions and compound structures should be understood to be within the scope described in the specification and / or claims of this application.

[0081] Unless otherwise specified, the numerical ranges described in the present specification and claims are equivalent to at least describing each specific integer value therein. For example, the numerical range "1-10" is equivalent to describing each integer value in the numerical range "1-10", 1, 2, 3, 4, 5, 6, 7, 8, 9, 10. It should be understood that when some numerical ranges are defined as "numbers", the two end points of the range, each integer in the range, and each decimal point in the range are described. For example, "numbers from 0 to 10" is equivalent to describing not only each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, but also at least the sum of each integer therein and each of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, and 0.9.

[0082] It should be understood that, when referring to one, two or more in this specification, "more" should refer to an integer greater than two, such as 3 or greater, for example 3, 4, 5, 6, 7, 8, 9 or 10.

[0083] The term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0084] "C 1-40 The term "alkyl group" should be understood to denote a linear or branched, saturated monovalent hydrocarbon group having 1 to 40 carbon atoms. For example, "C 1-10 "Alkyl group" refers to straight and branched chain alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms; 1-6The term "alkyl group" refers to straight-chain and branched-chain alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, an isopropyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an isopentyl group, a 2-methylbutyl group, a 1-methylbutyl group, a 1-ethylpropyl group, a 1,2-dimethylpropyl group, a neopentyl group, a 1,1-dimethylpropyl group, a 4-methylpentyl group, a 3-methylpentyl group, a 2-methylpentyl group, a 1-methylpentyl group, a 2-ethylbutyl group, a 1-ethylbutyl group, a 3,3-dimethylbutyl group, a 2,2-dimethylbutyl group, a 1,1-dimethylbutyl group, a 2,3-dimethylbutyl group, a 1,3-dimethylbutyl group, a 1,2-dimethylbutyl group, or the like, or an isomer thereof.

[0085] "C 3-10 The term "cycloalkyl group" should be understood to represent a saturated monovalent monocyclic, bicyclic (e.g., bridged, spiro) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. 3-10 The cycloalkyl group may be a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl; or a bicyclic hydrocarbon group such as bornyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, dicyclo[2.1.1]hexyl, dicyclo[2.2.1]heptyl, dicyclo[2.2.1]heptene, 6,6-dimethyldicyclo[3.1.1]heptyl, 2,6,6-trimethyldicyclo[3.1.1]heptyl, dicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonane, 2,6-diazaspiro[3,4]octane; or a tricyclic hydrocarbon group such as adamantyl.

[0086] Unless otherwise defined, the term "3- to 10-membered heterocyclyl group" refers to a saturated or unsaturated non-aromatic ring or ring system, for example a 4-, 5-, 6-, or 7-membered monocyclic, a 7-, 8-, 9-, or 10-membered bicyclic (e.g., fused, bridged, spiro) or a 10-membered tricyclic ring system, and containing at least one, for example 1, 2, 3, 4, 5, or more heteroatoms selected from O, S, and N, of which N and S may be optionally oxidized to various oxidation states, such as to form nitrogen oxide, -S(O)-, or -S(O)2- states. Preferably, the heterocyclyl group may be selected from "3- to 10-membered heterocyclyl groups". The heterocyclyl group may be linked to the remainder of the molecule by any one of the carbon atoms or the nitrogen atom (if present). The heterocyclyl group may include fused or bridged rings and spirocyclic rings. In particular, the heterocyclyl group may include, but is not limited to, a 4-membered ring such as azetidinyl or oxetanyl, a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, or pyrrolinyl, or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl, or a 7-membered ring such as diazepanyl. Optionally, the heterocyclyl group may be benzo-fused. The heterocyclyl group may be bicyclic, for example, but is not limited to, a 5,5-membered ring such as a hexahydrocyclopenta[c]pyrrol-2(1H)-yl ring, or a 5,6-membered bicyclic ring such as a hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The heterocyclyl group may be partially unsaturated, i.e., contain one or more double bonds, such as, but not limited to, a dihydrofuranyl group, a dihydropyranyl group, a 2,5-dihydro-1H-pyrrolyl group, a 4H-[1,3,4]thiadiazinyl group, a 4,5-dihydrooxazolyl group or a 4H-[1,4]thiazinyl group, or may be benzo-fused, such as, but not limited to, a dihydroisoquinolyl group.When the 3- to 10-membered heterocyclyl group is linked to another group to form a compound of the present disclosure, a carbon atom in the 3- to 10-membered heterocyclyl group may be linked to another group, or a hetero ring atom in the 3- to 10-membered heterocyclyl group ring may be linked to another group. For example, when the 3- to 10-membered heterocyclyl group is selected from a piperazinyl group, a nitrogen atom in the piperazinyl group may be linked to another group. Alternatively, when the 3- to 10-membered heterocyclyl group is selected from a piperidinyl group, a nitrogen atom in the piperidinyl group ring and a carbon atom at the para position thereof may be linked to another group.

[0087] "C 6-10 The term "aryl group" preferably denotes a monovalent aromatic or partially aromatic monocyclic or bicyclic (e.g. fused, bridged, spiro) hydrocarbon ring having from 6 to 10 carbon atoms, including monoaromatic rings or polyaromatic rings fused together, in particular rings having 6 carbon atoms, such as phenyl or biphenyl groups ("C6 aryl group"), or rings having 9 carbon atoms, such as indanyl or indenyl groups ("C9 aryl group"), or rings having 10 carbon atoms, such as naphthyl groups ("C 10 It should be understood that the above C may be an aryl group. 6-10 When the aryl group is substituted, it may be mono- or polysubstituted, and the substitution site is not limited, and may be, for example, substituted at the ortho, para, or meta position.

[0088] The term "5- to 10-membered heteroaryl group" should be understood to include monovalent monocyclic, bicyclic (e.g., fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 10 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, preferably 1 to 3 heteroatoms independently selected from N, O and S, and which may further be benzo-fused in each case. "Heteroaryl group" also refers to groups in which a heteroaryl ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the connecting anchor or point is on the heteroaryl ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7- or 8-indazinyl groups, 1-, 3-, 4-, 5-, 6- or 7-isoindolyl groups, 2-, 3-, 4-, 5-, 6- or 7-indolyl groups, 2-, 3-, 4-, 5-, 6- or 7-indazolyl groups, 2-, 4-, 5-, 6-, 7- or 8-purine groups, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-quinoazinyl groups, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-quinoazinyl groups, -, 6-, 7- or 8-quinolyl group, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl group, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl group, 2-, 3-, 4-, 5- or 6-naphthalidinyl group, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl group, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl group, 2-, 4-, 6- or 7-pteridyl group, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aHcarbazolyl group, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolylcarbazolyl group, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbolinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenanthridinyl group, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-acridinyl group, 1- , 2-, 4-, 5-, 6-, 7-, 8- or 9-dinyl group, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenothiazinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl group, 2-, 3-, 4-, 5-,6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzoisoquinolyl group, 2-, 3-, 4- or thieno[2,3-b]furanyl group, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazino[2,3-c]carbazolyl group, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]pyranyl group, 2-, 3-, 4-, 5-, 7- or 8- -5H-pyrido[2,3-d]-o-azinyl, 1-, 3- or 5-1H-pyrazolo[4,3-d]-thiazolyl, 2-, 4- or 5-4H-imidazo[4,5-d]thiazolyl, 3-, 5- or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5- or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8- or 9-furo[3,4-c]cinno linyl group, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10- or 11-4H-pyrido[2,3-c]carbazolyl group, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl group, 7-benzo[b]thienyl group, 2-, 4-, 5-, 6- or 7-benzoxazolyl group, 2-, 4-, 5-, 6- or 7-benzimidazolyl group, 2-, 4-, 4- , 5-, 6- or 7-benzothiazolyl group, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-benzoxapinyl group, 2-, 4-, 5-, 6-, 7- or 8-benzoazinyl group, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-1H-pyrrolo[1,2-b][2]benzoazapinyl group. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl groups, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl groups, 2-, 3-, 4-, 5-, 6- or 7-indolyl groups, 2-, 3-, 4-, 5-, 6- or 7-benzo[b]thienyl groups, 2-, 4-, 5-, 6- or 7-benzoxazolyl groups, 2-, 4-, 5-, 6- or 7-benzimidazolyl groups and 2-, 4-, 5-, 6- or 7-benzothiazolyl groups. When the above 5- to 10-membered heteroaryl groups are linked to other groups to form the compounds of the present disclosure, the carbon atoms in the 5- to 10-membered heteroaryl group ring may be linked to other groups,A heteroatom in the 5- to 10-membered heteroaryl ring may be linked to another group. When the 5- to 10-membered heteroaryl group is substituted, it may be mono- or polysubstituted. The substitution site is not limited, and for example, a hydrogen atom linked to a carbon atom in the heteroaryl ring may be substituted, or a hydrogen atom linked to a heteroatom in the heteroaryl ring may be substituted.

[0089] The term "spirocycle" refers to a ring system in which two rings share one ring-forming atom.

[0090] The term "fused ring" refers to a ring system in which two rings share two ring-forming atoms.

[0091] The term "bridged ring" refers to a ring system in which two rings share three or more ring-forming atoms.

[0092] Unless otherwise stated, heterocyclyl, heteroaryl or heteroarylene groups include all possible isomeric forms thereof, for example, positional isomers thereof.Thus, for some illustrative non-limiting examples, they may include one, two or more positions, such as 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-positions (if present), substituted or bonded to other groups, and include pyridin-2-yl, pyridinide-2-yl, pyridin-3-yl, pyridinide-3-yl, pyridin-4-yl and pyridinide-4-yl, thienyl or thienylene groups, including thien-2-yl, thien-3-yl and thienylene-3-yl, pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0093] The term "oxo" refers to an oxy substituent (=O) formed by oxidation of a carbon, nitrogen or sulfur atom in a substituent.

[0094] Unless otherwise stated, the definitions of terms in this specification apply equally to groups which contain the term, e.g., C 1-10 The definition of an alkyl group is C1-10 Alkyloxy group, C 3-10 This also applies to cycloalkyloxy groups.

[0095] Those skilled in the art will appreciate that the compounds of formula (I) can exist in various pharma- ceutically acceptable salt forms: if they contain a basic center, they can form acid addition salts; if they contain an acidic center, they can form base addition salts; and if they contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they can further form inner salts.

[0096] The compounds of the present disclosure may exist in the form of solvates (e.g., hydrates) in which the compounds of the present disclosure contain polar solvents, particularly, for example, water, methanol or ethanol, which are components of the crystal lattice of the compounds. The amount of polar solvent, particularly water, may be present in a stoichiometric or non-stoichiometric ratio.

[0097] The compounds of the present disclosure may be chiral due to their molecular structure, and therefore may exist in various enantiomeric forms. Thus, these compounds can exist in racemic or optically active forms. The compounds of the present disclosure include isomers or mixtures thereof, racemates, in which each chiral carbon is in the R or S configuration. The compounds of the present disclosure or intermediates thereof can be isolated into enantiomeric compounds or used in this form for synthesis by chemical or physical methods well known to those skilled in the art. In the case of racemic amines, diastereomers are produced from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as, for example, R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulfonic acids. Enantiomer resolution can also be achieved by chromatography using optically active resolving agents (e.g., dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers immobilized on silica gel). Suitable eluents for this purpose are solvent mixtures containing water or alcohols, e.g., hexane / isopropanol / acetonitrile.

[0098] The corresponding stable isomer can be isolated by known methods, such as by extraction, filtration, or column chromatography.

[0099] The term "patient" refers to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse or primate, and most preferably a human.

[0100] The term "therapeutically effective amount" refers to an amount of an active compound or drug that elicits a biological or medical response that a researcher, veterinarian, physician, or other clinician is looking for in a tissue, system, animal, individual, or human, including one or more of the following: (1) prevention of disease: e.g., preventing a disease, disorder, or condition in an individual susceptible to the disease, disorder, or condition, but not experiencing or manifesting the pathology or symptoms of the disease. (2) inhibition of disease: e.g., inhibiting a disease, disorder, or condition (i.e., preventing further progression of the pathology and / or symptoms) in an individual experiencing or manifesting the pathology or symptoms of the disease, disorder, or condition. (3) mitigation of disease: e.g., mitigating a disease, disorder, or condition (i.e., reversing the pathology and / or symptoms) in an individual experiencing or manifesting the pathology or symptoms of the disease, disorder, or condition. Therapeutically effective amounts can be initially estimated by cell culture assays, and initial doses can be estimated from in vivo data. Following these initial guidelines, one of skill in the art can determine effective doses for humans. It should be noted that standard pharmaceutical methods in cell cultures or experimental animals can be used to demonstrate, for example, LD 50 and ED 50 By measuring the toxicity and therapeutic efficacy of the compounds described herein, one can determine.

[0101] The term "disease or condition associated with LRRK kinase or LRRK2 kinase activity" refers to a disease or condition characterized by inappropriate kinase activity or overactivity of the kinase as defined herein. Inappropriate activity refers to (i) kinase expression in cells that do not normally express the kinase, (ii) increased kinase expression resulting in undesirable cell proliferation, differentiation and / or growth, or (iii) decreased kinase expression resulting in undesirable reduction in cell proliferation, differentiation and / or growth. Kinase overactivity refers to the amplification of a gene encoding a particular kinase or the occurrence of a certain level of kinase activity, which correlates with abnormal cell proliferation, differentiation and / or growth (i.e., as kinase levels increase, the severity of one or more symptoms of the cellular abnormality increases). Overactivity may be the result of ligand-independent or constitutive activation due to mutations, such as deletion of a fragment of the kinase responsible for ligand binding.

[0102] Examples of the "adjuvants" described in this disclosure are provided in the Handbook of Pharmaceutical Excipients, 2nd edition, 1994, edited by A Wade & PJ Weller.

[0103] The "carriers" or "diluents" described in this disclosure are well known in the pharmaceutical art and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (edited by AR Gennaro 1985).

[0104] The choice of drug carrier, adjuvant or diluent can be selected according to the desired route of administration and standard pharmaceutical practice. The pharmaceutical composition may or may not include any suitable binders, lubricants, suspending agents, coating agents, solubilizing agents, buffers, flavoring agents, surfactants, thickening agents, preservatives (including antioxidants) as carriers, adjuvants or diluents, as well as substances included to render the formulation isotonic with the blood of the subject.

[0105] Among them, pharmaceutical preparations in which the carrier is solid and which are adapted for oral administration are most preferably in unit dose dosage form, for example, pills, capsules or tablets each containing a predetermined amount of active compound. Tablets can be produced by pressing or molding, optionally with one or more accessory ingredients. Pressed tablets can be produced by compressing the active compound in a free-flowing form (e.g., powder or particles) in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, lubricating substance, surface active agent or dispersing agent. Molded tablets can be produced by molding the active compound and an inert liquid diluent in a mold. Tablets can be optionally coated, or if uncoated, can be optionally printed with a symbol. Capsules can be produced by filling the capsule shell with the active compound alone or in a mixture with one or more accessory ingredients, and sealing in the usual manner. Cachets are similar to capsules, in which the active compound, together with any accessory ingredients, is sealed in a rice paper film. The active compound can also be prepared into dispersible particles, which can be suspended in water or sprinkled on food before administration, for example. The particles may be packaged, for example, in a sachet, in which the carrier may be a liquid formulation suitable for oral administration, which may be a solution or suspension in an aqueous or non-aqueous liquid form, or may be presented as an oil-in-water liquid emulsion.

[0106] The term "pharmaceutically acceptable salts" includes suitable acid addition and alkali salts thereof. Suitable pharmaceutical salts are disclosed in J Pharm Sci, 66, 199, 1977, Berge et al. For example, salts may be formed using strong inorganic acids such as mineral acids such as hydrohalides (e.g., hydrochloric acid, hydrobromic acid, and hydroiodic acid), sulfuric acid, phosphoric sulfates, hydrogen sulfates, hemisulfates, thiocyanates, persulfates, and sulfonic acids; strong organic carboxylic acids such as alkane carboxylic acids having 1 to 4 carbon atoms, unsubstituted or substituted (e.g., by halogen), such as acetic acid; saturated or unsaturated dicarboxylic acids, such as oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, phthalic acid, and 4-phthalic acid; hydroxycarboxylic acids, such as ascorbic acid, ethanolic acid, lactic acid, malic acid, tartaric acid, and citric acid; amino acids, such as aspartic acid and glutamic acid; benzoic acid; and organic sulfonic acids, such as (C1-C4)-alkyl- or aryl-sulfonic acids, unsubstituted or substituted (e.g., by halogen), such as methane- and p-toluenesulfonic acid.

[0107] Suitable salts are, for example, acetate, trifluoroacetate, lactate, gluconate, citrate, tartrate, maleate, malate, pantothenate, adipate, alginate, aspartate, benzoate, butyrate, digluconate, cyclopentanecarboxylate, glucoheptonate, glycerophosphate, oxalate, heptanoate, caproate, fumarate, nicotinate, palmitate, pectinate, 3-phenylpropionate, picrate, neopentanoate, proline, glycerol, phosphate, phosphate-gluconate ... These include organic sulfonic acids such as pionate, tartrate, lactobionate, pivolate, camphorate, undecanoate and succinate, methanesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, camphorsulfonate, 2-naphthalenesulfonate, benzenesulfonate, p-chlorobenzenesulfonate and p-toluenesulfonate, and inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, hydrogen sulfate, hemisulfuric acid, thiocyanic acid, persulfuric acid, phosphoric acid and sulfonic acid.

[0108] The term "pharmaceutical acceptable ester" refers to the use of organic acids or alcohols / hydroxides to form esters with esterifiable functional groups in the structure of the compounds of the present disclosure. Organic acids include carboxylic acids such as alkane carboxylic acids having 1-12 carbon atoms, unsubstituted or substituted (e.g., by halogen), such as acetic acid, saturated or unsaturated dicarboxylic acids such as oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, phthalic acid, and 4-phthalic acid, hydroxycarboxylic acids such as ascorbic acid, ethanolic acid, lactic acid, malic acid, tartaric acid, and citric acid, amino acids such as aspartic acid and glutamic acid, benzoic acid, and organic sulfonic acids such as (C1-C4)-alkyl- or aryl-sulfonic acids, unsubstituted or substituted (e.g., by halogen), such as methane- and p-toluenesulfonic acid. Suitable hydroxides include inorganic hydroxides, such as sodium hydroxide, potassium hydroxide, calcium hydroxide, and aluminum hydroxide. Alcohols include alkanols having 1 to 12 carbon atoms which may be unsubstituted or substituted (eg, by a halogen).

[0109] The term "isotopically labeled compound" refers to at least one atom of a compound of the present disclosure that is replaced with an isotope. Examples of such isotopes include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, e.g., the corresponding 2 H, 3 H, 13 C. 14 C. 15 N, 17 O. 18 O. 31 P, 32 P, 35 S, 18 F and 36 Contains Cl. Deuterium (i.e. 2Substitution with isotopes such as H may be advantageous in some cases because it can provide certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dose requirements. For example, the present disclosure includes compounds of general formula (I) in which any one hydrogen atom is replaced by a deuterium atom.

[0110] The term "prodrug compound" refers to a covalently bonded compound that releases an active parent drug according to general formula (I) in vivo. Such prodrugs are generally compounds of the present disclosure in which one or more suitable groups have already been modified such that the modification can be reversed after administration to a human or mammalian subject. Although a second drug can be administered with such a prodrug for in vivo reversal, the reversal is generally performed by naturally occurring enzymes in such subjects. Examples of such modifications include the pharma- ceutically acceptable esters mentioned above, of which the reversal can be performed by esterases and the like.

[0111] The term "crystalline polymorphs" refers to various crystalline, polycrystalline and hydrated forms of the compounds of the present disclosure. It has been found in the pharmaceutical industry that chemical compounds can be isolated in any of these forms by purification and / or separation of the solvents used in the synthesis and preparation of such compounds.

[0112] The term "administration" denotes that the pharmaceutical composition of the present disclosure is applicable for rectal, intranasal, intrabronchial, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intraarterial and intradermal), intraperitoneal or intrathecal administration. Preferably, the formulation is an oral administration formulation. The formulation may conveniently be presented in unit dosage form, i.e., a form containing a unit dose or discrete portions of a unit dose or a multiple or sub-unit of a unit dose. Illustratively, the formulation may be in the form of tablets and sustained release capsules, and may be prepared by any method well known in the art of pharmacy.

[0113] Formulations of the present disclosure for oral administration may be presented as discrete units such as capsules, gels, lozenges, cachets, pills or tablets, each containing a predetermined amount of the active agent, as a powder or particles, as a solution, emulsion or suspension of the active agent in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as a bolus formulation, etc. Preferably, these compositions contain 1-250 mg, and more preferably 10-100 mg, of active ingredient per dose.

[0114] Compositions for oral administration (e.g., tablets and capsules) may further include solvents and / or commonly used adjuvants, such as syrup, gum arabic, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, binders such as sucrose and starch, fillers and carriers such as corn starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dibasic calcium phosphate, sodium chloride and alginic acid, and lubricants such as magnesium stearate, sodium stearate and other metal stearates, glyceryl stearate, stearic acid, silicone fluids, paraffin wax, oils and colloidal silica. Flavorings, such as mint, oil of wintergreen, cherry flavoring, and the like, may also be used. Colorings may need to be added to facilitate identification of the dosage form. Tablets may be coated by methods well known in the art.

[0115] Other formulations adapted for oral administration include topical coatings which generally contain a flavored matrix which is the active agent in sucrose and gum arabic or tragacanth, lozenges which contain the active agent in an inert matrix such as gelatin and glycerin, or sucrose and gum arabic, and mouthwashes which contain the active agent in a suitable liquid carrier.

[0116] Other administration forms include solutions or emulsions that are administered intravenously, intraarterially, intrathecally, subcutaneously, intradermally, intraperitoneally or intramuscularly and are prepared in a sterile or sterilizable solution. Injectable forms generally contain between 10 and 1000 mg, preferably between 10 and 250 mg, of active ingredient per formulation.

[0117] The above-mentioned administration modes may be combination administrations in which one or more compounds of the present disclosure are administered in combination with one or more other active agents, in which case the compounds of the present disclosure and one or more other active agents may be administered sequentially, simultaneously, or sequentially.

[0118] Analytical Testing Another aspect of the present disclosure relates to the use of the above-described compounds in assays used to identify other candidate compounds capable of inhibiting one or more kinases, more preferably LRRK, and even more preferably LRRK2.

[0119] Preferably, the assay is a competitive binding assay.

[0120] More preferably, the competitive binding assay comprises contacting a compound of the present disclosure with a kinase, preferably LRRK, more preferably LRRK2, and a candidate compound, and detecting any change in the interaction between the compound of the present disclosure and the kinase.

[0121] Preferably, candidate compounds are generated by SAR modification of a compound of the present disclosure.

[0122] As used herein, the term "SAR modification" refers to the standard method of altering a given compound by chemical derivatization.

[0123] Thus, in one embodiment, the identified compounds can be used as models (e.g., templates) for the development of other compounds. The compounds used in this assay can be free in solution, immobilized on a solid support, loaded on a cell surface, or intracellular. The formation of a binding complex or activity between the compound and the drug to be assayed can be measured.

[0124] The analytical test of the present disclosure may be a screen, so that many drugs are measured. In one aspect, the analytical measurement method of the present disclosure is a high throughput screen.

[0125] The present disclosure further contemplates the use of competitive drug screening assays in which neutralizing antibodies capable of binding to a compound specifically compete with the test compound for binding to the compound.

[0126] For high throughput screening (HTS) of reagents with suitable binding affinity to a substance, an alternative technique for screening is provided and is based on the method detailed in WO84 / 03564.

[0127] The analytical test method of the present disclosure is expected to be applicable to small-scale and large-scale screening of test compounds, as well as to quantitative testing.

[0128] Preferably, a competitive binding assay involves contacting a compound of the disclosure with a kinase in the presence of a known substrate of the kinase, and detecting any change in the interaction between the kinase and the known substrate.

[0129] Another aspect of the disclosure provides a method for detecting binding of a ligand to a kinase, the method comprising: (i) contacting a ligand with a kinase in the presence of a known substrate of said kinase; (ii) detecting any change in the interaction between the kinase and the known substrate; And wherein the ligand is a compound of the present disclosure.

[0130] One aspect of the present disclosure is (a) performing the measurement method; (b) identifying one or more ligands capable of binding to the ligand binding domain; (c) modifying said one or more ligands capable of binding to the ligand-binding domain; (d) performing such a measurement method; (e) optionally preparing a pharmaceutical composition comprising said one or more ligands; The present invention relates to a method comprising the steps of:

[0131] The present disclosure further relates to ligands identified by the above methods.

[0132] Another aspect of the disclosure relates to a pharmaceutical composition comprising a ligand identified by the above method.

[0133] Another aspect of the disclosure relates to the use of a ligand identified by the above method in the manufacture of a pharmaceutical composition, which is used for the treatment of one or more disease conditions.

[0134] The above method can be used to screen for ligands that can be used as inhibitors of one or more kinases.

[0135] BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is an exemplary diagram showing the Western blot results of LRRK2 expression levels in brain tissue from LRRK2-G2019S mice in Example 12.

[0136] FIG. 2 is a graph showing the effect of compound 527 of Example 12 on the phosphorylation level of LRRK2 in LRRK2-G2019S mouse brain tissue, where the data is the mean ± sample standard deviation (mean ± SE), n=8, * indicates p<0.05 compared to the solvent control group, *** indicates p<0.001 compared to the solvent control group, and single-factor ANOVA was used.

[0137] [Mode for carrying out the invention] The technical solution of the present disclosure will be described in more detail below in accordance with specific examples. It should be understood that the following examples are merely illustrative and explanatory of the present disclosure, and should not be construed as limiting the scope of the claims of the present disclosure. Any technology realized based on the above content of the present disclosure is included in the scope of the claims of the present disclosure.

[0138] Unless otherwise specified, all materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0139] In the context of this disclosure, the abbreviations have the following meanings: DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene DCM Dichloromethane DEA Diethylamine DIEA Diisopropylethylamine DMA N,N-Dimethylacetamide DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide EA, EtOAc, ethyl acetate IPA Isopropanol MTBE tert-butyl methyl ether NIS N-iodosuccinimide NMR nuclear magnetic resonance PE Petroleum Ether PMBCl p-Methoxybenzyl chloride R T, t R retention time SFC Supercritical Fluid Chromatography Tf Trifluoromethylsulfonyl group TfOH Trifluoromethanesulfonic acid TFA Trifluoroacetic acid THF Tetrahydrofuran UV ultraviolet light.

[0140] Chromatography High pressure liquid chromatography was used and monitored by a multi-wavelength UV detector. Typical mobile phases used in the separation process were PE / EA, DCM / MeOH or water / MeCN.

[0141] Analysis method Unless otherwise stated, measurements were performed using a Bruker AV 400 spectrometer in the above solvents at room temperature. 1 H nuclear magnetic resonance (NMR) spectroscopy was performed. In each case, the NMR data are consistent with the submitted structures. Common abbreviations are used to designate the main peaks and indicate characteristic chemical shifts (δ) in parts per million: e.g., s, singlet; d, doublet; t, triplet; q, quartet; dd, doublet of doublets; br, broad line. Mass spectra were recorded on an Agilent 1290 Infinity / 6460 triple Quad LCMS. When thin layer chromatography (TLC) was used, it refers to silica gel TLC.

[0142] Compound Production Where the preparation of starting materials is not described, these starting materials may be commercially available, known in the literature, or readily obtainable by one skilled in the art using standard procedures. Where the preparation of compounds is described by conventional examples or intermediates, it will be understood by those skilled in the art that the reaction times, number of equivalents of reagents and temperatures for each particular reaction may vary, and that the employment of different work-up or purification techniques may be necessary or desirable.

[0143] Example 1: Preparation of Compound 390

[0144] [ka]

[0145] Step 1: Synthesis of (2S,6R)-4-(6-chloropyrimidin-4-yl)-2,6-dimethylmorpholine (1-1) At room temperature, DIEA (87.7 g, 0.680 mol) and (2R,6S)-2,6-dimethylmorpholine (40.5 g, 0.350 mol) were added to a solution of 4,6-dichloropyrimidine (50.0 g, 0.340 mol) in IPA (1000 mL). The mixture was stirred at room temperature for 1 h. The solvent was removed by concentration, and the residue was diluted with EtOAc (500 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the residue. The residue was purified by silica gel column chromatography (PE / EA=1:1) to give a white solid (2S,6R)-4-(6-chloropyrimidin-4-yl)-2,6-dimethylmorpholine (1-1, 76.0 g, 98.4%). m / z (ESI) + : 228.0 [M+H] + .

[0146] Step 2: Synthesis of (2S,6R)-2,6-dimethyl-4-(6-(trimethylstannyl)pyrimidin-4-yl)morpholine (1-2) A dry three-neck flask containing xylene (100 mL) was heated to 150 °C under N2 atmosphere, then a solution of compound 1-1 (10.0 g, 44.1 mmol), a degassed mixture of 1,1,1,2,2,2-hexamethylstilbene (18.8 g, 57.3 mmol), Pd(PPh3)2Cl2 (3.00 g, 4.40 mmol) and PPh3 (2.30 g, 8.80 mmol) in xylene (50 mL) was added dropwise. The resulting mixture was stirred at 150 °C for 2 h. The reaction mixture was cooled to room temperature, diluted with PE and then filtered through diatomaceous earth. The filtrate was concentrated in vacuo to give the crude product (1-2, 16.0 g, ca. 60% purity) as a yellow solid, which was used in the next step without further purification. m / z (ESI) + : 358.1 [M+H] + .

[0147] Step 3: Synthesis of 4-chloro-3-iodo-1H-pyrazolo[4,3-c]pyridine (1-3) NIS (88.0 g, 0.390 mol) was added to a solution of 4-chloro-1H-pyrazolo[4,3-c]pyridine (50.0 g, 0.326 mol) in DMF (500 mL) at room temperature, and after the addition was complete, the mixture was heated to 80° C. and stirred for 2 h. The mixture was then concentrated to give a residue, which was treated with saturated aqueous NaHSO3 (500 mL) and stirred for 30 min. The solid formed was collected by filtration, washed with water (2 L), and then dried in vacuum to give the brown solid product (1-3, 83.0 g, 91.2%). m / z (ESI) + : 279.8 [M+H] + .

[0148] Step 4: Synthesis of 4-chloro-3-iodo-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridine (1-4) In an ice-water bath, PMBCl (49.6 g, 0.320 mol) and NaOH (23.2 g, 0.580 mol) were added to a solution of compound 1-3 (80.0 g, 0.290 mol) in DMF (800 mL). The mixture was stirred at room temperature for 20 h, concentrated to remove the solvent, and the residue was dissolved in EtOAc (500 mL), washed with brine, dried over anhydrous Na2SO4, and after filtration, the solution was concentrated to give the residue, which was slurried with PE and EA (PE / EA=1:1) and stirred overnight. Filtration and drying of the solid gave compound 1-4 (104 g, 89.7%) as an off-white solid. m / z (ESI) + : 399.7 [M+H] + .

[0149] Step 5: Synthesis of N-cyclopropyl-3-iodo-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (1-5) A mixture of compound 1-4 (7.00 g, 17.5 mmol) and cyclopropanamine (10.0 g, 175 mmol) in DIEA (50 mL) was heated to 140° C. and stirred for 2 days. It was concentrated in vacuo to remove the solvent, then diluted with EA and brine. The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (100% EA) to give compound 1-5 (5.00 g, 68.0%) in the form of a brown solid. m / z (ESI) + : 421.1 [M+H] + .

[0150] Step 6: Synthesis of N-cyclopropyl-3-(6-((2S,6R)-2,6-dimethylmorpholino)pyrimidin-4-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (1-6) To a mixture of compound 1-5 (200 mg, 0.476 mmol) and crude compound 1-2 (purity about 60%, 425 mg, 0.714 mmol) in DMF (10 mL) under N2 atmosphere, Pd(PPh3)2Cl2 (34 mg, 0.048 mmol), PPh3 (25 mg, 0.095 mmol) and CuI (9 mg, 0.043 mmol) were added. The mixture was degassed and purged with nitrogen gas three times, then stirred at 120 °C for 1 h. It was cooled to room temperature, diluted with EA, the organics were washed with saturated aqueous KF and brine, the organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (DCM / MeOH=19:1) to give compound 1-6 (50.0 mg, 21.6%) in the form of a light brown solid. m / z (ESI) + : 486.1 [M+H] + .

[0151] Step 7: Synthesis of N-(3,3-difluorocyclobutyl)-3-(6-(2S,6R)-2,6-dimethylmorpholino)pyrimidin-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (390) To a solution of compound 1-6 (50.0 mg, 0.103 mmol) in TFA (5 mL) was added TfOH (0.5 mL). The resulting mixture was stirred at room temperature for 1 h. It was concentrated to remove the solvent and diluted with DCM. It was then washed with 1 M aqueous NaOH and brine, the organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was purified by preparative TLC (DCM / MeOH=10:1) to give compound 390 (6 mg) as an off-white solid. 1 HNMR (400 MHz, CD3OD) δ 8.68 (d, J = 1.0 Hz, 1H), 7.72-7.65 (m, 2H), 7.10 (d, J = 7.2 Hz, 1H), 4.26-4.72 (m, 2H), 3.76-3.64 (m, 2H), 2.94-2.86 (m, 1H), 2.78-2.65 (m, 2H), 1.28 (d, J = 6.2 Hz, 6H), 1.20-1.13 (m, 2H), 0.95-0.88 (m, 2H). m / z (ESI) + : 366.0 [M+H] + .

[0152] With reference to Example 1, by using the corresponding starting materials in Step 5, the compounds shown in Table 1 were prepared.

[0153] [Table 1]

[0154] Example 2: Preparation of Compound 397

[0155] [ka]

[0156] Step 1: Synthesis of (cis)-2,6-dimethyltetrahydro-2H-pyran-4-ol (2-1) To a solution of 2,6-dimethyl-4H-pyran-4-one (5.00 g, 40.3 mmol) in ethanol (50 mL) was added Pd / C (10%, 0.500 g) and the reaction mixture was stirred at 35 °C and H2 (4 atm) for 20 h. The reaction mixture was filtered through diatomaceous earth and the filtrate was concentrated under reduced pressure to give compound 2-1 (5.00 g, 95.4%) in the form of pale yellow crystals. m / z (ESI) + : 131.1 [M+H] + .

[0157] Step 2: Synthesis of 1-isopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzaldehyde-2-yl)-1H-pyrazole (2-2) To a suspension of 4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzofuran-2-yl)-1H-pyrazole (5.00 g, 25.8 mmol) and Cs2CO3 (12.6 g, 38.6 mmol) in acetonitrile (100 mL) at room temperature was added 2-iodopropane (6.57 g, 38.6 mmol). The mixture was heated to 60° C. and stirred for 20 h. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA=4:1) to give colorless oily compound 2-2 (5.80 g, 95.2%). 1 HNMR (300 MHz, CDCl3) δ 7.78 (s, 1H), 7.73 (s, 1H), 4.51 (hept, J = 6.7 Hz, 1H), 1.49 (d, J = 6.7 Hz, 6H), 1.31 (s, 12H).

[0158] Step 3: Synthesis of 4-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)oxy)-3-iodo-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridine (2-3): At room temperature, NaH (60%) (187 mg, 4.68 mmol) was added to a solution of compound 2-1 (510 mg, 3.90 mmol) in DMA (10 mL). After stirring for 10 min, compound 1-4 (1.57 g, 3.9 mmol) was added. The resulting mixture was heated to 120° C. and stirred for 20 h. After cooling to room temperature, water (100 mL) was added and the resulting mixture was extracted with EtOAc (100 mL×2). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=3:2) to give compound 2-3 (680 mg, 35.3%) as a pale yellow oil. m / z (ESI) + : 493.7 [M+H] + .

[0159] Step 4: Synthesis of 4-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)oxy)-3-(1-isopropyl-1H-pyrazol-4-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridine (2-4): To a mixture of compound 2-3 (176 mg, 0.357 mmol), compound 2-2 (90 mg, 0.38 mmol) and Na2CO3 (121 mg, 1.14 mmol) in DMSO / H2O (5 mL / 0.5 mL) was added Pd(dppf)Cl2 (27 mg, 0.038 mmol). The mixture was degassed and purged with nitrogen gas three times, then stirred at 80 °C under nitrogen gas atmosphere for 20 h. After cooling to room temperature, EA and water were added. The organic layer was separated, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=32:3) to give compound 2-4 (100 mg, 59.0%) in the form of a light brown solid. m / z (ESI) + : 475.8 [M+H] + .

[0160] Step 5: Synthesis of 4-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)oxy)-3-(1-isopropyl-1H-pyrazol-4-yl)-1H-pyrazolo[4,3-c]pyridine (397): In a manner similar to step 7 of Example 1, 4-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)oxy)-3-(1-isopropyl-1H-pyrazol-4-yl)-1H-pyrazolo[4,3-c]pyridine (397) was synthesized from compound 2-4 to give a white solid. 1 HNMR (400 MHz, CD3OD) δ 8.31 (s, 1H), 8.14 (s, 1H), 7.86 (d, J = 6.1 Hz, 1H), 7.07 (d, J = 6.1 Hz, 1H), 5.54-5.45 (m, 1H), 4.67-4.59 (m, m / z (ESI) + : 355.8 [M+H] + .

[0161] With reference to Example 2, the compounds shown in Table 2 were prepared using the appropriate corresponding starting materials.

[0162] [Table 2]

[0163] Example 3: Preparation of Compounds 407 and 421

[0164] [ka]

[0165] Step 1: Synthesis of (2R,6S)-2,6-dimethyltetrahydro-4H-pyran-4-one (3-1) To a solution of 2,6-dimethyl-4H-pyran-4-one (100 g, 0.810 mol) in ethanol (800 mL) was added Pd / C (10%, 10 g), and the mixture was stirred at 30° C. and H2 (2 atm) for 8 h. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (PE / EA=3:1) to give crude compound 3-1 (53.0 g, 51.1%) as a colorless oil. m / z (ESI)+: 129.1 [m+H] + .

[0166] Step 2: Synthesis of (2R,6S)-2,6-dimethyltetrahydro-4H-pyran-4-one oxime (3-2) A suspension of compound 3-1 (15 g, 0.12 mol), hydroxylamine hydrochloride (8.1 g, 0.12 mol) and NaOAc (19.7 g, 0.24 mol) in ethanol (150 mL) was heated to 60 °C and stirred for 3 h. After cooling to room temperature, the mixture was filtered and the filtrate was concentrated in vacuum to give a residue. The residue was diluted with EtOAc and washed with brine. The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=3:7) to give compound 3-2 (16.0 g, 95.6%) as a white solid. m / z (ESI)+: 144.2 [m+H] + .

[0167] Step 3: Synthesis of (2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-amine (3-3) To a suspension of LiAlH4 (5.7 g, 0.15 mol) in THF (200 mL) was added dropwise a solution of compound 3-2 (7.2 g, 0.050 mol) at 0°C. The mixture was then heated to 60°C and stirred for 6 h. After cooling to 0°C in an ice-water bath, the reaction was quenched by sequential addition of water (5.7 mL), 15% aqueous NaOH (5.7 mL), and water (17.1 mL). Mg2SO4 was then added and stirred at room temperature for 1 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give compound 3-3 (6.10 g, 94.5%) as a colorless oil. m / z (ESI)+: 130.1[m+H] + .

[0168] Step 4: Synthesis of 4-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)oxy)-3-(1-isopropyl-1H-pyrazol-4-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridine (3-4) A mixture of compound 1-4 (18.6 g, 46.5 mmol) and compound 3-3 (6.00 g, 46.5 mmol) in DIEA (100 mL) was stirred at 140° C. for 36 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (DCM / MeOH=97:3) to give compound 3-4 (10.0 g, 43.7%) as a brown solid. m / z (ESI)+: 492.8[m+H] + .

[0169] Step 5: Synthesis of 3-(6-((2S,6R)-2,6-dimethylmorpholino)pyrimidin-4-yl)-N-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (3-5) Under nitrogen gas protection, Pd(PPh3)2Cl2 (280 mg, 0.400 mmol) and CuI (76 mg, 0.400 mmol) were added to a mixture of intermediate 3-4 (1.00 g, 2.00 mmol) and crude intermediate 1-2 (purity about 60%, 1.4 g, 2.35 mmol) in DMF (20 mL). The mixture was degassed and purged with nitrogen gas three times, then stirred at 120 °C for 2 h. After cooling to room temperature and diluting with EtOAc and water, the separated organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=1:9) to give intermediate 3-5 (300 mg, 26.9%) as a light brown solid. m / z (ESI)+: 557.7 [m+H] + .

[0170] Step 6: Synthesis of 3-(6-((2S,6R)-2,6-dimethylmorpholino)pyrimidin-4-yl)-N-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (407) and 3-(6-((2S,6R)-2,6-dimethylmorpholino)pyrimidin-4-yl)-N-((2R,4S,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (421) To a solution of intermediate 3-5 (300 mg, 0.54 mol) in TFA (20 mL) was added TfOH (2 mL). The mixture was stirred at room temperature for 1 h. It was concentrated, diluted with EtOAc, washed with 1 mol / L aqueous NaOH and brine, dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=1:9) and then separated by preparative HPLC (water, Gemini 5 μm, C18 150×21.2 mm, 20 mL / min, ACN / H2O (0.1% TFA)=33%:67%, 10 min hold) to give compound 407 (16.5 mg) RT=6.55 min and compound 421 (8.3 mg) RT=8.07 min as white solids.

[0171] Compound 407: 1 HNMR (400 MHz, CD3OD) δ 8.65 (s, 1H), 7.68 (s, 1H), 7.56 (d, J = 7.2 Hz, 1H), 7.03 (d, J = 7.2 Hz, 1H), 4.50-4.30 (m, 2H), 4.08-3.96 (m, m / z (ESI) + : 437.9 [M+H] + .

[0172] Compound 421: 1 HNMR (400 MHz, CD3OD) δ 8.55 (s, 1H), 7.74 (s, 1H), 7.59 (d, J = 7.2 Hz, 1H), 7.07 (d, J = 7.2 Hz, 1H), 4.63-4.36 (m, 2H), 4.32 (s, 1H), 4.01-3.93 (m, 2H), 3.73-3.64 (m, 2H), 2.70 (t, J = 11.8 Hz, 2H), 2.03-1.94 (m, 2H), 1.76-1.66 (m, 2H), 1.30-1.24 (m, 12H); m / z (ESI) + : 437.9 [M+H] + .

[0173] With reference to Example 1 and Example 3, the compounds in Table 3 were synthesized by using an appropriate amine instead of (2R,6S)-2,6-dimethylmorpholine in Step 1 of Example 1, and / or by using an appropriate amine instead of 3-3 in Step 4 of Example 3. Among them, compounds 527, 528, 561, and 562, which do not specify the retention time RT, were prepared by selecting chiral raw materials.

[0174] [Table 3] JPEG2025506822000021.jpg223169

[0175] Example 4: Preparation of Compound 422 and Compound 445

[0176] [ka]

[0177] Step 1: Synthesis of 4-iodo-1-isopropyl-1H-imidazole (4-1) To a solution of 4-iodo-1H-imidazole (20 g, 0.1 mol) and 2-iodopropane (21 g, 0.12 mol) in CH3CN (50 mL) was added Cs2CO3 (50 g, 0.15 mol). The resulting mixture was stirred at 60 °C overnight. It was cooled to room temperature, filtered, and the filtrate was concentrated. The residue was dissolved in EtOAc, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE / EA=1:1) to give compound 4-1 (20 g, 82%) as a colorless oil. m / z (ESI)+: 236.8 [m+H] + .

[0178] Step 2: Synthesis of (1-isopropyl-1H-imidazol-4-yl)boronic acid (4-2) Under nitrogen gas protection, a solution of n-butyllithium (n-BuLi) in n-hexane (2.4 M, 16 mL, 38 mmol) was slowly added dropwise to a solution of compound 4-1 (3 g, 13 mmol) and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxybenzaldehyde (7.1 g, 38 mmol) in THF (20 mL) at -75 °C. After the addition was completed, the mixture was stirred at -75 °C for 2 h. The reaction was quenched by adding NH4Cl (saturated aqueous solution, 50 mL). Extraction with EtOAc was performed, and the organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to obtain a residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 10:1) to obtain compound 4-2 (1.7 g, 87%) as a white solid. m / z (ESI)+: 155.2 [m+H] + .

[0179] Step 3: Synthesis of N-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-imidazol-4-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (4-3) N-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-imidazol-4-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (4-3) was synthesized from compounds 4-2 and 3-4 in a manner similar to step 4 of Example 2, and was a white solid. m / z (ESI)+: 475.2 [m+H] + .

[0180] Step 4: Synthesis of N-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-imidazol-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (422) and N-((2R,4S,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-imidazol-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (445) To a solution of intermediate 4-3 (200 mg, 0.42 mol) in TFA (10 mL) was added TfOH (0.5 mL). The mixture was stirred at room temperature for 1 h. The mixture was concentrated, diluted with EtOAc, washed with 1 M aqueous NaOH and brine, dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=1:9) and then separated by SFC (chiral chromatography column chiralpak IC column, 4.6 mm×250 mm, 5 μm, eluted with CO2-EtOH (0.2% NH4OH)) to give compound 422 (9 mg) RT=15.6 min and compound 445 (3 mg) RT=10.09 min as white solids.

[0181] Compound 422: 1 HNMR (400 MHz, CD3OD) δ 7.95 (s, 1H), 7.79 (s, 1H), 7.59 (d, J = 6.6 Hz, 1H), 6.76 (d, J = 6.6 Hz, 1H), 4.60-4.52 (m, 1H), 4.17-4.07 (m, m / z (ESI) + : 355.2 [M+H] + .

[0182] Compound 445: 1HNMR (400 MHz, CD3OD) 7.88 (s, 1H), 7.84 (s, 1H), 7.61 (d, J = 6.6 Hz, 1H), 6.78 (d, J = 6.6 Hz, 1H), 4.65-4.55 (m, 1H), 4.45-4.37 (m, m / z (ESI) + : 355.2 [M+H] + .

[0183] With reference to Examples 3 and 4, by substituting the appropriate amine for amine 3-3 in step 4 of Example 3, the compounds shown in Table 4 can be prepared.

[0184] [Table 4]

[0185] Example 5: Preparation of Compounds 423 and 439

[0186] [ka]

[0187] Step 1: Synthesis of 1-isopropyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxybenzaldehyde-2-yl)-1H-pyrazole (5-1) 3-(4,4,5,5-tetramethyl-1,3,2-dioxybenzofuran-2-yl)-1H-pyrazole (1.04 g, 5.36 mmol) was added to a suspension of NaH (60%, 0.24 g, 6 mmol) in DMF (10 mL) under nitrogen gas atmosphere at 0°C. The mixture was stirred at 30°C for 30 min. Then it was cooled to 0°C and 2-iodopropane (0.8 mL, 8 mmol) was added dropwise. The resulting mixture was stirred at room temperature for another 30 min. The reaction was quenched by adding saturated aqueous NH4Cl to the mixture, which was then extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to give the residue. The residue was purified by silica gel column chromatography (PE / EA=4:1) to give compound 5-1 (550 mg, 43%) as a colorless oil. 1 HNMR (300 MHz, CD3OD) δ 7.67 (d, J = 2.3 Hz, 1H), 6.59 (d, J = 2.3 Hz, 1H), 4.66-4.52 (m, 1H), 1.48 (d, J = 6.7 Hz, 6H), 1.19(s, 12H).

[0188] Step 2: Synthesis of N-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-pyrazol-3-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (5-2): N-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-imidazol-4-yl)-1-(4-methoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (5-2) was synthesized from compounds 5-1 and 3-4 in a manner similar to step 4 of Example 2, and was a white solid. m / z (ESI) + : 475.2 [M+H] + .

[0189] Step 3: Synthesis of N-((2R,4R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-pyrazol-3-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (423) and N-((2R,4S,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-3-(1-isopropyl-1H-pyrazol-3-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (439) To a solution of intermediate 5-2 (200 mg, 0.42 mmol) in TFA (10 mL) was added TfOH (0.5 mL). The mixture was stirred at room temperature for 1 h. The mixture was concentrated, diluted with EtOAc, washed with 1 M aqueous NaOH and brine, dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by silica gel column chromatography (DCM / MeOH=10:1) and then separated by SFC (chiral chromatography column chiralpak IC column, 4.6 mm×250 mm, 5 μm, eluted with CO2-IPA (0.2% NH4OH)) to give compound 423 (50 mg) as a white solid, RT=10.55 min, and compound 439 (17 mg), RT=7.90 min.

[0190] Compound 423: 1 HNMR (400 MHz, CD3OD) δ 7.86 (d, J = 2.4 Hz, 1H), 7.61 (d, J = 6.9 Hz, 1H), 6.95-6.90 (m, 2H), 4.76-4.66 (m, 1H), 4.21-4.13 (m, 1H), 3.77-3.68 (m, 2H), 2.28-2.21 (m, 2H), 1.64 (d, J = 6.7 Hz, 6H), 1.44-1.33 (m, 2H), 1.29 (d, J = 6.2 Hz, 6H); m / z (ESI) + : 354.9 [M+H] + .

[0191] Compound 439: 1HNMR (400 MHz, CD3OD) δ 7.87 (d, J = 2.5 Hz, 1H), 7.70 (d, J = 6.4 Hz, 1H), 6.96 (d, J = 2.5 Hz, 1H), 6.76 (d, J = 6.4 Hz, 1H), 4.67-4.57 (m, 1H), 4.54-4.49 (m, 1H), 4.06-3.97 (m, 2H), 2.04-1.97 (m, 2H), 1.67-1.62 (m, 2H), 1.60 (d, J = 6.8 Hz, 6H), 1.23 (d, J = 6.2 Hz, 6H); m / z (ESI) + : 354.9 [M+H] + .

[0192] With reference to Examples 3 and 5, by substituting the appropriate amine for amine 3-3 in step 4 of Example 3, the compounds shown in Table 5 can be prepared.

[0193] [Table 5]

[0194] Example 6: Preparation of Compound 523

[0195] [ka]

[0196] Step 1: Synthesis of tert-butyl 2-(6-chloropyrimidin-4-yl)-2-cyanoacetate (6-1) Sodium hydride (60%, 6.71 g, 0.168 mol) was added to a solution of 4,6-dichloropyrimidine (10 g, 0.067 mol) and tert-butyl 2-cyanoacetate (23.7 g, 0.168 mol) in THF (100 mL) at 0°C under nitrogen gas. The reaction mixture was stirred at room temperature for 2 h. The mixture was quenched by adding saturated aqueous NH4Cl solution, and then the pH value was adjusted to 2-3 with 1 M aqueous HCl solution. Extracted twice with EtOAc, the combined organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to give compound 6-1 (16 g, 94%) as a white solid. m / z(ESI)+: 253.9[m+H] + .

[0197] Step 2: Synthesis of 2-(6-chloropyrimidin-4-yl)acetonitrile (6-2) To a solution of intermediate 6-1 (11 g, 0.043 mol) in toluene (100 mL) was added p-toluenesulfonic acid monohydrate (0.82 g, 0.0043 mol). The mixture was stirred at 100° C. for 3 h. Cooled to room temperature, diluted with EtOAc, the mixture was washed with saturated aqueous NaHCO3 and brine, dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=7:3) to give compound 6-2 (5.5 g, 84%) as a yellow solid. m / z(ESI)+: 154.0[m+H] + .

[0198] Step 3: Synthesis of 1-(6-chloropyrimidin-4-yl)cyclobutane-1-carbonitrile (6-3) In an ice-water bath, a solution of intermediate 6-2 (612 mg, 4 mmol) and 1,3-dibromopropane (965 mg, 4.8 mmol) in DMF (10 mL) was added dropwise to a suspension of potassium carbonate (1.38 g, 10 mmol) in DMF (10 mL). The reaction mixture was stirred at room temperature for 3 h. The mixture was diluted with EtOAc, then washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA=1:1) to give compound 6-3 (600 mg, 78%) as a yellow solid. m / z(ESI)+: 194.0 [m+H] + .

[0199] Step 4: Synthesis of 1-(6-(trimethylstannyl)pyrimidin-4-yl)cyclobutane-1-carbonitrile (6-4) 1-(6-(trimethylstannyl)pyrimidin-4-yl)cyclobutane-1-carbonitrile (6-4) was synthesized from compound 6-3 in a manner similar to step 2 of Example 1, and was a yellow solid. m / z(ESI)+: 323.9 [m+H] + .

[0200] Step 5: Synthesis of 3-iodo-1-(4-methoxybenzyl)-N-(2,2,2-trifluoroethyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (6-5) A suspension of intermediate 1-4 (30 g, 0.075 mol), 2,2,2-trifluoroethylamine hydrochloride (30.5 g, 0.225 mol) and DBU (23 g, 0.15 mmol) in xylene (250 mL) was sealed and heated to 190 °C in an oil bath and stirred for 3 h. It was cooled to room temperature, diluted with EtOAc, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to give a residue. The residue was stirred with MTBE at room temperature for 30 min and filtered to give compound 6-5 (24 g, 69%) as a white solid. m / z(ESI)+: 463.1 [m+H] + .

[0201] Step 6: Synthesis of 1-(6-(1-(4-methoxybenzyl)-4-(2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)pyrimidin-4-yl)cyclobutane-1-carbonitrile (6-6) 1-(6-(1-(4-Methoxybenzyl)-4-(2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)pyrimidin-4-yl)cyclobutane-1-carbonitrile (6-6) was synthesized from compounds 6-4 and 6-5 in a manner similar to step 5 of example 3, yellow solid. m / z(ESI)+: 493.9[m+H]+.

[0202] Step 7: Synthesis of 1-(6-(4-((2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)pyrimidin-4-yl)cyclobutane-1-carbonitrile (523) 1-(6-(4-((2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)pyrimidin-4-yl)cyclobutane-1-carbonitrile (523) was synthesized from compound 6-6 in a manner similar to step 6 of example 3, as a yellow solid. 1 HNMR (400 MHz, DMSO-d6) δ 13.65 (s, 1H), 10.26 (s, 1H), 8.08 (s, 1H), 7.80 (d, J = 6.0 Hz, 1H), 7.32 (s, 1H), 6.80 (d, J = 6.0 Hz, 1H), 4.48-4.35 (m, 2H), 3.92-3.81 (m, 2H), 2.34-2.27 (m, 2H), 1.90-1.80 (m, 2H); m / z (ESI) + : 373.8 [M+H] + .

[0203] By referring to Example 6 and substituting the appropriate halogenated hydrocarbon for 1,3-dibromopropane in Step 2, or substituting the appropriate amine for 2,2,2-trifluoroethylamine in Step 5, the compounds shown in Table 6 were prepared.

[0204] [Table 6]

[0205] Example 7: Preparation of Compounds 521 and 509

[0206] [ka]

[0207] Step 1: Synthesis of 2-(4,4,5,5-tetramethyl-1,3,2-dioxybenzaldehyde-2-yl)-1H-pyrazol-1-yl)acetonitrile (7-1) Sodium hydride (60%, 4.12 g, 0.103 mol) was added to a solution of 4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzofuran-2-yl)-1H-pyrazole (10 g, 0.052 mol) in DMF (150 mL) while stirring at 0°C. After stirring for 30 min, 2-bromoacetonitrile (12.35 g, 0.103 mol) was added. The mixture was stirred at room temperature overnight. The mixture was quenched by adding saturated aqueous NH4Cl solution and extracted twice with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by silica gel column chromatography (DCM / MeOH=97:3) to give compound 7-1 (2.0 g, 17%) as an off-white solid. m / z (ESI)+: 234.0[m+H] + .

[0208] Step 2: Synthesis of 5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzaldehyde-2-yl)-1H-pyrazol-1-yl)spiro[2.3]hexane-5-carbonitrile (7-2) Under nitrogen gas protection, a solution of intermediate 7-1 (2.05 g, 8.8 mmol) and 1,1-bis(bromomethyl)cyclopropane (2.0 g, 8.8 mmol) in THF (50 mL) was added dropwise to a solution of LiHMDS (1 M, 43 mL, 43 mmol) in THF (150 mL) at -5°C to 0°C. The mixture was stirred at that temperature for 10 minutes, and then the reaction was quenched with formic acid. The mixture was concentrated to obtain a residue. The residue was purified by silica gel column chromatography (DCM / MeOH=99:1) to obtain compound 7-2 (170 mg, 26%) as a colorless oil. m / z (ESI)+: 300.0 [m+H] + .

[0209] Step 3: Synthesis of 5-(4-(1-(4-methoxybenzyl)-4-(2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-1H-pyrazol-1-yl)spiro[2.3]hexane-5-carbonitrile (7-3) 5-(4-(1-(4-methoxybenzyl)-4-(2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-1H-pyrazol-1-yl)spiro[2.3]hexane-5-carbonitrile (7-3) was synthesized from compounds 7-2 and 6-5 in a manner similar to step 4 of example 2, and was a yellow solid. 1 HNMR (300 MHz, CDCl3) δ 8.04 (s, 1H), 7.96 (s, 1H), 7.87 (d, J = 6.1 Hz, 1H), 7.20-7.15 (m, 2H), 6.86-6.79 (m, 2H), 6.68 (d, J = 6.1 Hz, 1H), 5.43 (s, 2H), 5.16 (t, J = 6.3 Hz, 1H), 4.37-4.24 (m, 2H), 3.76 (s, 3H), 3.34-3.26 (m, 2H), 3.03-2.95 (m, 2H), 0.85-0.77 (m, 2H), 0.67-0.59 (m, 2H); m / z (ESI) + : 507.7 [M+H] + .

[0210] Step 4: Synthesis of 3-ethylidene-1-(4-(4-((2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (521) and 5-(4-((2,2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-1H-pyrazol-1-yl)spiro[2.3]hexane-5-carbonitrile (509) In a manner similar to step 7 of Example 1, 1-(6-(4-((2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)pyrimidin-4-yl)cyclobutane-1-carbonitrile (521) and 5-(4-(4-((2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-1H-pyrazol-1-yl)spiro[2.3]hexane-5-carbonitrile (509) were synthesized from compound 7-3 as white solids.

[0211] Compound 521: RT = 8.3 min (preparative HPLC (GILSON, Gemini 5 μm, C18 150 x 21.2 mm, 20 mL / min, ACN / H2O (0.05%NH4OH) = 35%:65%~60%:40%, within 10 min)); 1 HNMR (400 MHz, CD3OD) δ 8.26 (s, 1H), 7.99 (s, 1H), 7.84 (d, J = 6.2 Hz, 1H), 6.89 (d, J = 6.2 Hz, 1H), 5.60-5.52 (m, 1H), 4.28 (q, J = 9.5 Hz, 2H), 3.79-3.64 (m, 4H), 1.66-1.60 (m, 3H); m / z (ESI) + : 387.8 [M+H] + .

[0212] Compound 509: RT = 7.7 min (preparative HPLC (GILSON, Gemini 5 μm, C18 150 x 21.2 mm, 20 mL / min, ACN / H2O (0.05%NH4OH) = 35%:65%~60%:40%, within 10 min)); 1 HNMR (400 MHz, CD3OD) δ 8.30 (s, 1H), 7.99 (s, 1H), 7.84 (d, J = 6.2 Hz, 1H), 6.89 (d, J = 6.2 Hz, 1H), 5.60-5.52 (m, 1H), 4.29 (q, J = 9.5 Hz, 2H), 3.30-3.26 (m, 2H), 3.04-2.98 (m, 2H), 0.83-0.77 (m, 2H), 0.66-0.59 (m, 2H); + : 387.9 [M+H] + .

[0213] Example 8: Preparation of Compound 526

[0214] [ka]

[0215] Step 1: Synthesis of 3-(6-((1R,5S)-6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)pyrimidin-4-yl)-1-(4-methoxybenzyl)-N-(2,2,2-trifluoroethyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (8-1) 3-(6-((1R,5S)-6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)pyrimidin-4-yl)-1-(4-methoxybenzyl)-N-(2,2,2-trifluoroethyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (8-1) was synthesized from 4,6-dichloropyrimidine in a manner similar to that described in Example 1 using the appropriate reagents at the appropriate steps.

[0216] Step 2: Synthesis of (3S,5R)-1-(6-(4-((2,2,2-trifluoroethyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)pyrimidin-4-yl)piperidine-3,5-diol (526) To a solution of compound 8-1 (90 mg, 0.176 mmol) in TFA (10 mL) was added TfOH (0.5 mL). The resulting mixture was stirred at room temperature for 1 h, then concentrated, diluted with DCM, and then washed with 1 M aqueous NaOH and brine. The separated organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by preparative TLC (DCM / MeOH=5:1) and preparative HPLC (Gilson, Gemini 5 μm, C18 150×21.2 mm, 20 mL / min, ACN / H2O (0.1% FA)=35%:65%~50%:50% in 10 min) to give compound 526 (20 mg) as a white solid. 1 HNMR (400 MHz, CD3OD) δ 8.58 (d, J = 1.0 Hz, 1H), 7.77 (d, J = 6.3 Hz, 1H), 7.71 (d, J = 1.0 Hz, 1H), 6.87 (d, J = 6.3 Hz, 1H), 4.39 (q, J = m / z (ESI) + : 410.0 [M+H] + .

[0217] Example 9: Preparation of Compound 522

[0218] [ka]

[0219] Step 1: Synthesis of (2R,6S)-4-(2-bromopyridin-4-yl)-2,6-dimethylmorpholine (9-1) Under nitrogen atmosphere, cuprous iodide (70 mg, 0.352 mmol) and L-proline (80 mg, 0.704 mmol) were added to a suspension of 2-bromo-4-iodopyridine (1.0 g, 3.52 mmol), (2R,6S)-2,6-dimethylmorpholine (400 mg, 3.52 mmol) and K2CO3 (1.5 g, 10.5 mmol) in DMSO (20 mL). The resulting mixture was degassed and purged with nitrogen gas three times, then stirred at 60 °C for 1 h. The reaction mixture was diluted with EtOAc and washed with brine. The separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give the residue. The residue was purified by silica gel column chromatography (DCM / MeOH=99:1) to give compound 9-1 (400 mg, 42%) as a white solid. m / z (ESI)+: 270.8, 272.8 [m+H] + .

[0220] Step 2: Synthesis of 3-(4-((2S,6R)-2,6-dimethylmorpholino)pyridin-2-yl)-N-(2,2,2-trifluoroethyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (522) 3-(4-((2S,6R)-2,6-dimethylmorpholino)pyridin-2-yl)-N-(2,2,2-trifluoroethyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (522) was synthesized from compound 9-1 in a manner similar to that described in Example 1 using the appropriate reagents at the appropriate steps. 1 HNMR (400 MHz, CDCl3) δ 8.25 (d, J = 6.1 Hz, 1H), 7.87 (d, J = 6.1 Hz, 1H), 7.76 (d, J = 2.6 Hz, 1H), 6.71 (dd, J = 6.1, 2.6 Hz, 2H), 4.50-4.36 (m, 2H), 3.81-3.71 (m, 4H), 2.66-2.58 (m, 2H), 1.30 (d, J = 6.2 Hz, 6H. m / z (ESI) + : 406.8 [M+H] + .

[0221] Production Example: Synthesis of Reference Compound

[0222] [ka]

[0223] The above control compound was synthesized by referring to the method of Example 1, with the difference being that step 1 was not carried out, step 2 was carried out using 2-chloropyridine, the compound 1-2 in Example 1 was replaced with the prepared 2-(trimethylstannyl)pyridine, and CF3CH2NH2 was used instead of cyclopropanamine in step 5 to obtain the control compound.

[0224] Example 10. Measurement of kinase inhibition material Kinase LRRK2 G2019S protein was from Carna, LRRKtide was from Signalchem, ATP was from Promega, and DMSO was from Sigma.

[0225] Kinase Test Method All measurements were performed at room temperature. Compounds to be measured were dissolved and diluted, then added to a 384-well plate, 2.5 μL of 2× kinase reaction buffer was added, centrifuged, 2.5 μL of 2× substrate and ATP mix was added, centrifuged, 4 μL of ADP-Glo ​​reagent was added, and incubated at room temperature for 40 min. 8 μL of LRRKtide was added, incubated at room temperature for 40 min, and then the data was read using an Envision 2104 multi-label Reader.

[0226] result IC of compounds of the examples of the present disclosure for LRRK2 G2019S inhibition 50 The values ​​are shown in Table 3 below.

[0227] [Table 7]

[0228] Example 11. Measurement of HEK293 cell kinase inhibition HEK293 cells were cultured, pancreatin was added to harvest the cells, and medium was added to bring the cell density to 2 × 10 5 The liposome-DNA complex was constructed, mixed with HEK293 cells at 1:20, added to a 384-well plate, and incubated at 37°C for 20-30 hours. A mixture of enzyme substrate and compound to be measured was added, incubated at room temperature for 2-3 minutes, NanoBRET reagent (Promega) was added, and the bioluminescence of the donor luciferase protein (donor) and the acceptor fluorescent group (acceptor) at 450 nm and 610 nm, respectively, were read within 10 minutes using a microplate reader (Envision 2104). The BRET ratio of the test product including the background correction option was calculated according to the following formula:

[0229]

number

[0230] The inhibitory activity of the compounds to be measured against the cellular LRRK2 enzyme was calculated according to the following formula:

[0231] Percent inhibition=100−(test article BRET ratio−total microplate positive control average BRET ratio) / (total microplate negative control average BRET ratio−total microplate positive control average BRET ratio)×100.

[0232] IC of pending compounds against cellular LRRK2 enzyme 50 Calculation of: Nonlinear regression fitting (dose-response slope method) was performed on the percent inhibition and log concentration curves of the compounds using Graphpad software. Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)×Hill Slope)) X: log inhibitor concentration, Y: percent inhibition, Bottom: minimum value, Top: maximum value, Hill Slope: Hill slope.

[0233] result IC of some example compounds of the present disclosure for inhibition of LRRK2, LRRK2-2019S and LRRK2-R1441C in HEK293 cells 50 The values ​​are summarized in the table below.

[0234] [Table 8]

[0235] Example 12: In vivo efficacy study by oral administration to mice 1. Experimental Objective: The in vivo efficacy of compound 527 was measured by observing the inhibitory effect of oral administration of compound 527 on the LRRK2 phosphorylation level in the brain of mice with the G2019S mutant LRRK2 gene knocked in.

[0236] 2. Experimental Method: (1) Animals: B6.Cg-Tg(Lrrk2*G2019S)2Yue / J transgenic mice were purchased from Jackson Laboratory and crossed with C57BL / 6J mice to obtain LRRK2 heterozygous mice carrying the G2019S mutation (LRRK2-G2019S mice). Half male and half female, weighing 16–20 g, were 15–17 weeks old.

[0237] (2) Grouping and administration: Each group had 8 animals, 4 female mice and 4 male mice. The mice were randomly grouped into 5 experimental groups based on body weight. The specific administration of the experimental groups is shown in Table 1 below. Among them, group G1 was the solvent control group, and groups G2 to G5 were administered with 527 compound at doses of 1 mg / kg, 3 mg / kg, 10 mg / kg and 30 mg / kg, respectively.

[0238] [Table 9]

[0239] (3) Measurement of LRRK2 activity in the brain: 1 h after administration, the animals were euthanized with CO2, and brain tissue was collected and rapidly frozen in liquid nitrogen, then frozen at -80C until the next experiment. TM Brain tissue was homogenized in RIPA buffer containing protease inhibitor mixture (Roche, 04693124001) and phosphatase enzyme inhibitor mixture 2 (Sigma, R0278), and protein was quantified using the BCA method. Western blot was used to detect the phosphorylated LRRK2 level at pS935 (primary antibody Anti-LRRK2 (phospho S935) antibody, Abcam, ab133450) and total LRRK2 level (primary antibody Anti-LRRK2 antibody, Abcam, ab133518) in the homogenized tissue, and the brain LRRK2 activity level was expressed as the phosphorylation level at S935 of LRRK2, i.e., the ratio of pS935-LRRK2 to total LRRK2 (pS935-LRRK2 / LRRK2).

[0240] (4) Results and conclusions: Typical results of brain tissue Western blot are shown in FIG. 1, and the inhibitory effect of compound 527 on the phosphorylation of brain tissue LRRK2 is shown in FIG.

[0241] The results showed that the effective dose of compound 527 for inhibiting brain tissue LRRK2 phosphorylation by oral administration was 10 mg / kg, the maximum pharmacological dose (defined as inhibiting LRRK2 phosphorylation level by 60%) was 30 mg / kg, and the corresponding human equivalent dose (calculated based on a body weight of 70 kg) was 57 mg and 171 mg, respectively.

[0242] The exemplary embodiments of the present disclosure have been described above. However, the scope of the claims of the present disclosure is not limited to the above embodiments. Any modifications, equivalent replacements, or improvements made by those skilled in the art without departing from the spirit and principles of the present disclosure are included in the scope of the claims of the present disclosure. [Brief description of the drawings]

[0243] [Figure 1]FIG. 13 is an exemplary diagram showing the western blot results of LRRK2 expression levels in brain tissue of LRRK2-G2019S mice in Example 12. [Diagram 2] 1 is a graph showing the effect of compound 527 of Example 12 on the phosphorylation level of LRRK2 in LRRK2-G2019S mouse brain tissue, in which the data is the mean ± sample standard deviation (mean ± SE), n = 8, * represents p < 0.05 compared to the solvent control group, *** represents p < 0.001 compared to the solvent control group, and single-factor ANOVA is used.

Claims

1. A compound represented by formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester or a prodrug compound thereof, 【Chemistry 1】 Among them, W is selected from N or O; R 1 is unsubstituted or contains one, two or more R a C replaced with 1-10 Alkyl group, C 3-10 cycloalkyl groups, and 3- to 10-membered heterocyclyl groups; Each R a are identical or different and independently represent a halogen, a hydroxyl group, C 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, and a 3- to 10-membered heterocyclyloxy group; Or, alternatively, two R a are unsubstituted or optionally substituted with one, two or more R c C replaced with 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 forming a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, or a 3- to 10-membered heterocyclyloxy group; R 2 is absent or H or C 1-10 alkyl groups, in which when W is selected from O, R 2 does not exist and W is chosen from N, then R 2 is H or C 1-10 selected from alkyl groups, Or, alternatively, R 1 , R 2 is unsubstituted or optionally one, two or more R b forming a 3- to 10-membered heterocyclyl group substituted with Each R b are identical or different and independently represent a halogen, a hydroxyl group, C 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, and a 3- to 10-membered heterocyclyloxy group; Or, alternatively, two R b are unsubstituted or optionally substituted with one, two or more R c C replaced with 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 forming a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, or a 3- to 10-membered heterocyclyloxy group; Each R c are identical or different and independently represent a halogen, a hydroxyl group, C 1-10 Alkyl group, C 1-10 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, and a 3- to 10-membered heterocyclyloxy group; X is one, two or more R 3 5-10 membered heteroaryl groups substituted with Each R 3 are identical or different and are independently unsubstituted or substituted with one, two or more R 4 C replaced with 1-10 Alkyl group, C 3-10 selected from a cycloalkyl group, a 3- to 10-membered heterocyclyl group, and a 5- to 10-membered heteroaryl group; Or, alternatively, two R 3 are unsubstituted or optionally substituted with one, two or more R 4 C replaced with 3-10 forming a cycloalkyl group or a 3- to 10-membered heterocyclyl group; Each R 4 are identical or different and independently represent a halogen, a hydroxyl group, a cyano group, no substitution or one, two or more R d C replaced with 1-10 Alkyl group, C 1-10 Alkylene group, C 3-10 A cycloalkyl group, a 3- to 10-membered heterocyclyl group, and a 5- to 10-membered heteroaryl group, among which R 4 C 1-6 When selected from alkylene groups, R 4 The carbon atom in is R 3 means to form a double bond with the carbon atom linked to Each R d are identical or different and each independently represents a halogen, a hydroxyl group, a cyano group, C 1-6 Alkyl group, C 1-6 Alkyloxy group, C 3-10 Cycloalkyl groups, C 3-10 selected from a cycloalkyloxy group, a 3- to 10-membered heterocyclyl group, a 3- to 10-membered heterocyclyloxy group, a 5- to 10-membered heteroaryl group, and a 5- to 10-membered heteroaryloxy group; A compound represented by formula (I), its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester or prodrug compound thereof.

2. Each R a are identical or different and independently represent F, Cl, Br, C 1-6 selected from alkyl groups, R 1 is unsubstituted or contains one, two or more R a C replaced with 1-8 Alkyl group, C 3-8 cycloalkyl groups, and 3- to 8-membered heterocyclyl groups, e.g., R 1 is unsubstituted or contains one, two or more R a C replaced with 1-6 Alkyl group, C 3-7 cycloalkyl groups, and 3- to 7-membered heterocyclyl groups, e.g., R 1 teeth, 【Chemistry 2】 -CH 2 CF 3 , -CH(CH 3 )CF 3 Selected from Or, R 1 , R 2 is unsubstituted or optionally one, two or more R b Replaced with 【Chemistry 3】 Forming X is one, two or more R 3 For example, X is selected from one, two or more R 3 Replaced with 【Chemistry 4】 For example, X is selected from 【Chemistry 5】 Selected from Each R 4 are identical or different and independently represent fluorine, chlorine, bromine, a hydroxyl group, a cyano group, unsubstituted or one, two or more R d C replaced with 1-6 Alkyl group, C 1-6 Alkylene group, C 3-8 cycloalkyl groups, and 3- to 8-membered heterocyclyl groups; R 3 is unsubstituted or contains one, two or more R 4 C replaced with 1-6 Alkyl group, C 3-8 selected from a cycloalkyl group, a 3- to 8-membered heterocyclyl group, and a 5- to 6-membered heteroaryl group; For example, R 3 is unsubstituted or contains one, two or more R 4 methyl, ethyl, propyl, isopropyl, cyclopropane, cyclobutane substituted with 【Chemistry 6】 For example, R 3 is an isopropyl group, 【Chemistry 7】 Selected from The compound according to claim 1, its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester or prodrug compound thereof.

3. The compound of formula (I) has the structure of formula (II): 【Chemistry 8】 Among them, R 5 is C substituted with one, two or more halogens 1-6 Alkyl groups, preferably C substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 halogens 1-6 alkyl groups, for example methyl or ethyl groups substituted by 1, 2, 3, 4 or 5 fluorine or chlorine atoms, for example trifluoromethyl groups, R 6 is C 1-6 alkyl groups, such as methyl, ethyl, propyl or isopropyl groups; R 7 , R 8 , R 9 , R 10 are homologous or different and independently represent H or C 1-6 an alkyl group, for example selected from H, methyl, ethyl, propyl or isopropyl; R 11 , R 12 are homologous or different and independently represent H or C 1-6 Alkyl groups, for example selected from H, methyl, ethyl, propyl or isopropyl groups; The compound according to claim 1 or 2, its racemate, stereoisomer, tautomer, isotope-labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester or prodrug compound thereof.

4. The compound represented by formula (I) includes a compound represented by the following formula (III): 【Chemistry 9】 Among them, R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 has the definition set forth in claim 3, The compound according to claim 3, its racemate, stereoisomer, tautomer, isotope-labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma-ceutically acceptable ester or prodrug compound thereof.

5. The compound represented by formula (I) is 【Chemistry 10】 【change】 【change】 Selected from The compound according to claim 1, its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester or prodrug compound thereof.

6. A method for producing the compound according to any one of claims 1 to 5, its racemate, stereoisomer, tautomer, isotope-labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester, or a prodrug compound thereof, comprising the steps of: The method includes the following reaction: 【Chemistry 11】 Among them, R 1 , R 2 , W and X have the definitions described above; P 1 is selected from amino protecting groups, Preferably, the compound of formula (M2) is provided with a protecting group P 1 to obtain a compound represented by formula (I), Preferably, the P 1 is selected from alkyloxycarbonyl groups (e.g., tert-butoxycarbonyl groups, etc.), cycloalkylalkoxycarbonyl groups (e.g., cyclohexylmethoxycarbonyl groups), substituted alkyloxycarbonyl groups (e.g., trichloroethoxycarbonyl groups, etc.), substituted arylalkyloxycarbonyl groups (e.g., p-nitrobenzyloxycarbonyl groups), arylalkyl groups (e.g., triphenylmethyl groups or diphenylmethyl groups), heteroarylalkyl groups (e.g., pyridylalkyl groups, e.g., 2-pyridylmethyl groups, 4-pyridylmethyl groups), and alkoxyarylalkyl groups (e.g., alkoxyphenylalkyl groups, e.g., p-methoxybenzyl groups); Preferably, the process further comprises the following reaction: 【Chemistry 12】 Among them, R 1 , R 2 , W, X and P 1 has the definition set forth above, L 1 and L 2 are identical or different and are selected from leaving groups which enable coupling reactions independently of one another, Preferably, the compound of formula (M2) can be prepared by coupling reactions known in the art, for example by Stille coupling reaction or Suzuki coupling reaction, For example, L 1 may be selected from halogens such as iodine, L 2 may be selected from an alkyl tin group or a borate ester group; A manufacturing method comprising:

7. 1. A pharmaceutical composition comprising: The pharmaceutical composition comprises at least one of the compounds according to any one of claims 1 to 5, their racemates, stereoisomers, tautomers, isotopically labeled compounds, N-oxides, hydrates, solvates, crystalline polymorphs, metabolites, pharma- ceutically acceptable salts, pharma- ceutically acceptable esters, or prodrug compounds thereof; Preferably, the pharmaceutical composition further comprises one or more pharma- ceutically acceptable adjuvants, Preferably, the pharmaceutical compositions are in unit dosage form, each unit dosage containing from about 1 to 1000 mg, for example, from about 5 to 1000 mg, more typically from about 5 to 500 mg, for example, from 5 to 100 mg, 100 to 500 mg or 50 to 200 mg of the active ingredient; Preferably, the dosage range of the compound, its racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester or prodrug compound thereof is from about 1 μg / kg to about 1 g / kg body weight / day, e.g., from 0.01 mg / kg to about 100 mg / kg body weight / day; If the subject is a human, the dosage administered to the patient may be between 57 mg / 70 kg and 171 mg / 70 kg; A pharmaceutical composition comprising:

8. Use of at least one of the compounds according to any one of claims 1 to 5, their racemates, stereoisomers, tautomers, isotopically labeled compounds, N-oxides, hydrates, solvates, crystalline polymorphs, metabolites, pharma- ceutically acceptable salts, pharma- ceutically acceptable esters or prodrug compounds thereof in the manufacture of a medicament, comprising: The drug is used for treating a disease or condition associated with LRRK kinase, particularly LRRK2 kinase activity.

2. The use according to claim 1 ,

9. Use of at least one of the compounds according to any one of claims 1 to 5, their racemates, stereoisomers, tautomers, isotopically labeled compounds, N-oxides, hydrates, solvates, crystalline polymorphs, metabolites, pharma- ceutically acceptable salts, pharma- ceutically acceptable esters or prodrug compounds thereof in an analytical measurement, comprising: said assay being used to identify compounds capable of inhibiting one or more kinases, said kinase being preferably LRRK, more preferably LRRK2; Preferably, the analytical assay is a competitive binding assay. use.

10. 1. A method for detecting binding of a compound to a kinase, comprising: (i) contacting at least one of the compounds of any one of claims 1 to 5, or a racemate, stereoisomer, tautomer, isotopically labeled compound, N-oxide, hydrate, solvate, crystalline polymorph, metabolite, pharma- ceutically acceptable salt, pharma- ceutically acceptable ester, or a prodrug compound thereof, with a kinase in the presence of the kinase and a known substrate; (ii) detecting any change in the interaction between the kinase and the known substrate; method.

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