Amino acid derivatives, pharmaceutical compositions, their preparation methods and applications

Abstract

This invention relates to amino acid derivatives, pharmaceutical compositions, methods of preparation thereof, and applications thereof. Specifically, it provides a compound represented by Formula I, which has good LRRK2 inhibitory activity and can be used to treat or prevent LRRK2-related symptoms and diseases, as well as to prepare drugs for such symptoms and diseases.

Description

[0001] This invention claims priority to an earlier application filed on April 6, 2022, with the China National Intellectual Property Administration, patent application number 202210360017.2, entitled "Amino Acid Derivatives, Pharmaceutical Compositions and Preparation Methods Thereof and Applications Thereof," the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of drug synthesis, specifically relating to amino acid derivatives, pharmaceutical compositions, their preparation methods and applications. Background Technology

[0003] Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease. Its main pathological features are the degeneration and loss of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies. Clinically, PD presents with motor symptoms such as resting tremor, bradykinesia, muscle rigidity, and postural instability, often accompanied by sleep disturbances, autonomic dysfunction, psychiatric symptoms, and cognitive impairment. The pathogenesis of PD is not fully understood, but it is generally believed to be the result of multiple factors, with environmental and genetic factors working together to ultimately lead to its development. The complexity and insidious nature of the disease often prevent a definitive diagnosis in its early stages, leading to uncontrollable symptoms in its terminal stages. The role of genes in the pathogenesis of Parkinson's disease has gradually attracted attention. With the maturity of gene sequencing technology, gene detection methods are widely used in the diagnosis of Parkinson's disease. To date, more than 20 pathogenic genes have been identified, including leucine-rich repeat kinase 2 (LRRK2). LRRK2, also known as PARK8, is located on chromosome 12q12, is approximately 144kb long, contains 51 exons, and encodes 2527 amino acids. It is composed of protein domains such as LRR, Ras protein complex (ROC), C-terminal repeat sequence of Ras protein complex (COR), kinase active region (MAPKKK), and WD40. ROC and COR have protein kinase functions, and WD40 is a repeat sequence composed of 40 tryptophan and aspartic acid residues. This region can participate in the interaction of LRRK2 with other proteins and can also reversibly bind to proteins, participating in their signal transduction and transport. The LRRK2 protein, in conjunction with GTP and protein kinases, is widely expressed in various tissues and participates in central nervous system functions such as the substantia nigra, striatum, and caudate nucleus. LRRK2 gene mutations are a common cause of autosomal dominant Parkinson's disease and are also associated with sporadic Parkinson's disease. Currently, over 80 missense mutations have been identified, distributed throughout the entire LRRK2 gene segment and affecting all predicted functional domains. Preclinical studies have found that LRRK2-mutant Parkinson's disease is associated with the loss of substantia nigra neurons, along with the formation of Lewy bodies, suggesting that the LRRK2 protein may contribute to Parkinson's disease along a common pathway with α-synuclein. Further research has shown that abnormally elevated LRRK2 kinase activity can lead to a 3-4 fold increase in kinase activity, resulting in dopaminergic neuronal damage and significantly increasing the risk of Parkinson's disease. Clinical studies have also found that Parkinson's disease patients carrying the LRRK2 variant gene may experience faster progression of motor severity.Currently, scientists have discovered that LRRK2 kinase inhibitors can protect neurons from neurodegeneration induced by LRRK2 overactivation, block the accumulation of increased α-synuclein in neurons, inhibit inclusion body formation, and slow disease progression. Therefore, LRRK2 is considered a novel and highly promising target for the treatment of Parkinson's disease. Clinical trials targeting LRRK2-specific therapies for LRRK2 mutation carriers have begun, highlighting the rapid progress made in this field over the past decade. Progress in ongoing clinical studies has also demonstrated that LRRK2 inhibitors hold promise as potential disease-modifying therapies for improving LRRK2-Parkinson's disease. The kinase activity of LRRK2 is important for pathogenesis, and the LRRK2 kinase domain modulates overall LRRK2 function. Despite progress in central nervous system research on LRRK2, there is still a need for inhibitors of the LRRK2 receptor suitable for treating various neurodegenerative diseases, such as Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS). This invention provides a novel amino acid derivative structure and has found that compounds with this structure exhibit good activity. Summary of the Invention

[0004] To address the aforementioned technical problems, the present invention, in a first aspect, provides a compound of Formula I, its racemic mixture, stereoisomer, tautomer, isotope label, solvate, polymorph, pharmaceutically acceptable salt, or prodrug thereof:

[0005]

[0006] Wherein, R1 is selected from unsubstituted or optionally substituted by one, two or more R1s. 11 The following groups are substituted: NH2, C 6-20 Aryl, 5-20 quinone heteroaryl; each R 11 Same or different, selected independently from C 1-40 Alkyl, Halogenated C 1-40 Alkyl, C 3-20 cycloalkyl, diC 1-40 alkyl-phosphoryl-C 6-20 Aryl;

[0007] R2 is selected from C 1-40 Alkyl, C 1-40 Alkoxy, halogenated C 1-40 Alkyl groups, halogens;

[0008] R3 is selected from H and C. 1-40 Alkyl, C 1-40 Alkoxy;

[0009] Alternatively, R1, R2, and the atoms they are connected to form a group consisting of one, two, or more R atoms. 21 The following groups are substituted: C3-20 Cycloalkyl, 3-20 membered heterocyclic, 5-20 membered heteroaryl; each R 21 They may be the same or different, and are independently selected from halogens, CN, oxo (=O), and C. 1-40 Alkyl, C 3-20 cycloalkyl;

[0010] Alternatively, R2, R3, and the atoms they are connected to form a group consisting of one, two, or more R atoms. 22 The following groups are substituted: C 3-20 Cycloalkyl, 3-20 membered heterocyclic, 5-20 membered heteroaryl; each R 22 They may be the same or different, and are independently selected from halogens, CN, oxo (=O), and C. 1-40 Alkyl, C 3-20 cycloalkyl;

[0011] X is selected from N or CH;

[0012] R4 is selected from H, halogen, CN, OH, unsubstituted or optionally surrounded by one, two or more R4 groups. 41 The following groups are substituted: C 1-40 Alkyl, C 2-40 alkenyl, C 2-40 alkynyl group, C 3-20 cycloalkyl, C 1-40 Alkoxy, C 3-20 Cycloalkyloxy; each R 41 Whether the same or different, they are independently selected from deuterium, halogens, and carbon. 1-40 Alkyl, Halogenated C 1-40 Alkyl, C 3-20 cycloalkyl;

[0013] And / or, X and one of R4, together with the ring atoms they are attached to and the imino group (NH), form a 5-6 membered heterocyclic group;

[0014] m is selected from integers from 0 to 6; for example, 1, 2, 3, 4, 5;

[0015] Ring A is selected from C 6-20 Aryl, 5-20 heteroaryl;

[0016] E is selected from chemical bonds or R E1 Selected from C 1-40 Alkyl, C 1-40 Alkoxy, halogenated C 1-40 Alkyl groups, halogens; G is selected from chemical bonds, C 6-20 Aryl or 5-20 heteroaryl; n is selected from 0, 1, 2, 3 or 4;

[0017] R5, R6, R7, and R8 may be the same or different, and are independently selected from H and C.1-40 Alkyl, C 1-40 Alkoxy, NH2, NH2-C 1-40 Alkyl groups may be linked together to form a ring, or connected to one of the R4 groups to form a C group fused with ring A. 3-20 cycloalkyl, C 3-20 Cycloalkenyl, C 3-20 Cycloalkynyl, 3-20 membered heterocyclic, 5-20 membered heteroaryl, or R5, R6 and the atoms attached to them form C 3-20 Cycloalkyl, 3-20 membered heterocyclic group; and at least one of R5, R6, R7, and R8 is NH2 or NH2-C. 1-40 alkyl;

[0018] Y is selected from OR a 、N(R b (R) c ); R a Selected from H, C 1-40 alkyl or R b R c They are either the same or different, and are independently selected from H and C. 1-40 alkyl.

[0019] According to an embodiment of the invention, R1 is selected from unsubstituted or optionally replaced by one, two or more R1s. 11 The following groups are substituted: NH2, C 6-14 Aryl; each R 11 Same or different, selected independently from C 1-12 Alkyl, Halogenated C 1-12 Alkyl, C 3-12 cycloalkyl, diC 1-12 alkyl-phosphoryl-C 6-14 Aryl;

[0020] R2 can be selected from C. 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl groups, halogens;

[0021] R3 can be selected from H and C. 1-12 Alkyl, C 1-12 Alkoxy;

[0022] Alternatively, R1, R2, and the atoms they are connected to form a group consisting of one, two, or more R atoms. 21 The following groups are substituted: C 3-12 Cycloalkyl, 3-14 membered heterocyclic, 5-14 membered heteroaryl; each R 21 They may be the same or different, and are independently selected from halogens, CN, oxo (=O), and C. 1-12 Alkyl, C 3-12 cycloalkyl;

[0023] Alternatively, R2, R3, and the atoms they are connected to form a group consisting of one, two, or more R atoms. 22 The following groups are substituted: C 3-12 Cycloalkyl, 3-14 membered heterocyclic, 5-14 membered heteroaryl; each R 22 They may be the same or different, and are independently selected from halogens, CN, oxo (=O), and C. 1-12 Alkyl, C 3-12 Cycloalkyl.

[0024] According to an embodiment of the invention, R1 is selected from unsubstituted or optionally replaced by one, two or more R1s. 11 The following groups are substituted: NH2, phenyl; each R 11 Same or different, selected independently from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl, diC 1-6 alkyl-phosphoryl-phenyl;

[0025] R2 can be selected from halogenated C. 1-6 Alkyl groups, halogens;

[0026] R3 can be H;

[0027] Alternatively, R1, R2, and the atoms they are connected to form a group consisting of one, two, or more R atoms. 21 Substituted 3-8 membered heterocyclic groups, 5-8 membered heteroaryl groups; each R 21 They are either the same or different, and are independently selected from halogens, oxygen (=O), and carbon. 1-6 Alkyl, C 3-6 cycloalkyl;

[0028] Alternatively, R2, R3, and the atoms they are connected to form a group consisting of one, two, or more R atoms. 22 Substituted 3-8 membered heterocyclic groups, 5-8 membered heteroaryl groups; each R 22 They may be the same or different, and are independently selected from halogens, CN, oxo (=O), and C. 1-6 Alkyl, C 3-6 Cycloalkyl.

[0029] According to an embodiment of the present invention, R1 is selected from methylamino, ethylamino, propylamino, isopropylamino, cyclopropylamino, cyclobutylamino, difluoroethylamino, trifluoroethylamino, tolyl, and dimethylphosphorylphenylamino.

[0030] R2 can be selected from trifluoromethyl, F, Cl, Br;

[0031] Alternatively, R1, R2, and the atoms they are connected to form a group consisting of one, two, or more R atoms.21 Substituted tetrahydropyrrole; each R 21 They may be the same or different, and are independently selected from F, oxo (=O), and cyclopropyl groups; for example...

[0032] Alternatively, R2, R3, and the atoms they are connected to form a group consisting of one, two, or more R atoms. 22 The following groups are substituted: tetrahydropyrrolyl, 1H-pyrrolyl; each R 22 They may be the same or different, and are independently selected from F, CN, oxo (=O), and cyclopropyl; for example...

[0033] According to embodiments of the present invention, R4 is selected from H, halogens, CN, unsubstituted or optionally replaced by one, two or more Rs. 41 The following groups are substituted: C 1-12 Alkyl, C 2-12 alkynyl group, C 3-12 cycloalkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyloxy; each R 41 Whether the same or different, they are independently selected from deuterium, halogens, and carbon. 1-12 Alkyl, Halogenated C 1-12 Alkyl, C 3-12 cycloalkyl;

[0034] And / or, X and one of R4, together with the ring atoms they are attached to and the imino group (NH), form a 5-6 membered heterocyclic group;

[0035] And / or, R5 and one of R4 are connected to form a C that fused with ring A. 3-12 cycloalkyl, C 3-12 Cycloalkenyl, 3-12 heterocyclic or 5-12 heteroaryl.

[0036] According to an embodiment of the present invention, R4 is selected from H, halogens, CN, and C. 1-12 Alkyl, C 2-12 alkynyl group, C 3-12 cycloalkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyloxy, halogenated C 1-12 Alkoxy, deuterated C 1-12 Alkoxy;

[0037] And / or, X and one of R4, together with the ring atoms they are attached to and the imino group (NH), form a 5-6 membered heterocyclic group;

[0038] And / or, R4 and R5 are connected to form a 5-6 membered heterocyclic group fused with ring A.

[0039] According to an embodiment of the present invention, R4 is selected from H, F, Cl, Br, I, CN, and C. 1-6 Alkyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyloxy, halogenated C 1-6 Alkoxy, deuterated C 1-6 Alkoxy;

[0040] And / or, X and one of R4, together with the ring atoms they are attached to and the imino group (NH), form a 6-membered heterocyclic group;

[0041] And / or, R5 and one of R4 are connected to form a 5-membered heterocyclic group or a 5-membered heteroaryl group fused with ring A.

[0042] According to an embodiment of the present invention, R4 is selected from H, F, Cl, CN, methyl, methoxy, ethoxy, cyclopropyl, cyclopropyloxy, difluoromethoxy, trifluoromethoxy, trideuterated methoxy, and ethynyl.

[0043] And / or, X and one of R4, together with the ring atoms they are attached to and the imino group (NH), form a morpholino group;

[0044] And / or, R4 and R5 are connected to form a ring fused with ring A.

[0045] According to an embodiment of the present invention, ring A is selected from C. 6-14 Aryl, 5-14 heteroaryl; for example, selected from C 6-8 Aryl, 5-8 membered heteroaryl; such as those selected from phenyl, pyridyl, thiophene, pyrazolyl.

[0046] According to an embodiment of the present invention, E is selected from chemical bonds or R E1 Selected from fluorine, chlorine, bromine, and iodine; G is selected from chemical bonds or C. 6-14 Aryl; n is selected from 1 or 2; preferably, when n is 1, R E1 It is an R configuration;

[0047] E, for example, is selected from chemical bonds, Preferably, Selected from

[0048] According to an embodiment of the present invention, R5, R6, R7, and R8 may be the same or different, and are independently selected from H and C. 1-12 Alkyl, NH2 or NH2-C 1-12 Alkyl group; and at least one of R5, R6, R7, and R8 is NH2 or NH2-C. 1-12 alkyl;

[0049] Alternatively, R5, R6 and the atoms they are connected to form C 3-12 Cycloalkyl, 3-12 membered heterocyclic groups.

[0050] According to an embodiment of the present invention, R5, R6, R7, and R8 may be the same or different, and are independently selected from H and C. 1-6 Alkyl, NH2 or NH2-C 1-6 Alkyl, such as H, methyl, NH2, NH2-methyl; and at least one of R5, R6, R7, R8 is NH2 or NH2-methyl, for example, when R5, R6, and R7 are all H, R8 is NH2; when R5, R7, and R8 are all H, R6 is NH2 or NH2-methyl; when R5 and R6 are both H, R7 is methyl and R8 is NH2; when R5 and R7 are both H, R6 is methyl or ethyl and R8 is NH2; preferably, when R8 is NH2, it is an S configuration;

[0051] Alternatively, R5, R6 and the atoms they are connected to form C 3-6 Cycloalkyl groups, such as cyclopropyl groups.

[0052] According to an embodiment of the present invention, Y is selected from OR a 、N(R b (R) c ); R a Selected from H, C 1-12 alkyl or R b R c Same or different, selected independently from C 1-12 alkyl.

[0053] According to an embodiment of the present invention, Y is selected from OR a 、N(R b (R) c ); R a Selected from H, C 1-6 alkyl or R b R c Same or different, selected independently from C 1-6 Alkyl; Y is selected, for example, from OH, methylamino, dimethylamino, isopropyloxy, tert-butyloxy, heptayloxy, or

[0054] According to embodiments of the present invention, the compound shown in Formula I may be selected from the structures shown in Formula II or Formula III:

[0055]

[0056] Among them, R1, R2, R3, R4, R5, R6, R7, R8, E, Y and m independently have the definitions described above.

[0057] According to embodiments of the present invention, the compound shown in Formula I may be selected from the structures shown in Formula I-1, I-2, I-3 or I-4:

[0058]

[0059] Among them, R1, R4, R5, R6, G and Y independently have the definitions described above.

[0060] According to a preferred embodiment of the present invention, the compound of representative formula I according to the present invention has a structure selected from the following:

[0061]

[0062]

[0063]

[0064] The present invention also provides a method for preparing a compound of formula I, comprising at least one of the following schemes:

[0065] Option 1: Deprotection of compound I-1 yields the compound shown in Formula I;

[0066]

[0067] Option 2: Deprotection of compound I-2 yields the compound shown in Formula I;

[0068]

[0069] Among them, R1, R2, R3, R4, R5, R6, R7, R8, m, A, E, X, and Y independently have the definitions described above, and PG is selected from amino protecting groups, such as tert-butoxycarbonyl and benzyloxycarbonyl.

[0070] According to an embodiment of the present invention, the deprotection reaction in Scheme 1 or Scheme 2 can be carried out in the presence of an acid or a catalyst, wherein the acid is preferably selected from HCl solution or trifluoroacetic acid; and the catalyst is preferably selected from palladium on carbon or palladium dioxide.

[0071] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the following: a compound of formula I, a racemic mixture, a stereoisomer, a tautomer, an isotope label, a solvate, a polymorph, a pharmaceutically acceptable salt, or a prodrug thereof.

[0072] According to embodiments of the present invention, the pharmaceutical composition further includes one or more pharmaceutically acceptable excipients.

[0073] According to embodiments of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0074] In a third aspect, the present invention provides the use of compounds of Formula I, their racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrugs thereof in the preparation of pharmaceuticals, for example, in the preparation of LRRK2 inhibitors.

[0075] According to embodiments of the present invention, the drug or drug composition is used for the prevention or treatment of diseases or symptoms mediated by LRRK2.

[0076] According to embodiments of the present invention, the diseases or symptoms associated with LRRK2 mediation are selected from Parkinson's disease, leprosy, IBD, Alzheimer's disease, L-DOPA-induced dyskinesia, dementia, amyotrophic lateral sclerosis, kidney cancer, breast cancer, prostate cancer, leukemia, papillary carcinoma, lung cancer, acute myeloid leukemia, multiple myeloma, leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis, or ankylosing spondylitis.

[0077] According to the present invention, compounds of Formula I, their racemates, stereoisomers, tautomers, isotope-labeled forms, solvates, polymorphs, pharmaceutically acceptable salts, or prodrugs thereof can be formulated into forms suitable for administration via any appropriate route, using conventional methods with one or more pharmaceutically acceptable carriers. Therefore, compounds of Formula I, their racemates, stereoisomers, tautomers, isotope-labeled forms, solvates, polymorphs, pharmaceutically acceptable salts, or prodrugs thereof can be formulated into various dosage forms for oral administration, injection (e.g., intravenous, intramuscular, or subcutaneous) administration, inhalation, or blow-through administration; they can also be formulated into sustained-release dosage forms, such as tablets, hard or soft capsules, aqueous or oily suspensions, emulsions, injections, dispersible powders or granules, suppositories, lozenges, or syrups.

[0078] The present invention also provides a method for treating or preventing LRRK2-mediated diseases, comprising administering to a patient a preventive or therapeutically effective amount of at least one of the following: a compound of Formula I, its racemic, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound, or a pharmaceutical composition thereof.

[0079] In some implementations, the patient is a mammal, preferably a human.

[0080] The present invention also provides at least one of the following: a compound of Formula I, a racemic mixture, a stereoisomer, a tautomer, an isotope label, a solvate, a polymorph, a pharmaceutically acceptable salt, or a prodrug compound thereof, or a pharmaceutical composition thereof, for the use of in the treatment or prevention of diseases mediated by LRRK2.

[0081] According to embodiments of the present invention, the diseases or symptoms associated with LRRK2 mediation are selected from Parkinson's disease, leprosy, IBD, Alzheimer's disease, L-DOPA-induced dyskinesia, dementia, amyotrophic lateral sclerosis, kidney cancer, breast cancer, prostate cancer, leukemia, papillary carcinoma, lung cancer, acute myeloid leukemia, multiple myeloma, leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis, or ankylosing spondylitis.

[0082] Beneficial effects

[0083] The compounds provided by this invention have good LRRK2 inhibitory effects and can be used to treat or prevent LRRK2-related conditions and diseases, as well as to prepare drugs for such conditions and diseases.

[0084] Terminology Definitions and Explanations

[0085] Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures should be understood as being within the scope of this application specification and / or claims.

[0086] Unless otherwise stated, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-40" is equivalent to describing each integer value in the numerical range "1-10", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and each integer value in the numerical range "11-40", namely 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40. Furthermore, when certain numerical ranges are defined as "numbers", it should be understood that they describe the two endpoints of the range, each integer within the range, and each decimal within the range. For example, "numbers from 0 to 10" should be understood as not only recording each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also recording at least the sum of each of these integers with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 and 0.9 respectively.

[0087] It should be understood that in the description of 1, 2 or more, "more" should refer to an integer greater than 2, such as an integer greater than or equal to 3, such as 3, 4, 5, 6, 7, 8, 9 or 10.

[0088] The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0089] Term "C" 1-40 "Alkyl" should be understood as referring to a straight-chain or branched saturated monovalent hydrocarbon group having 1 to 40 carbon atoms. For example, "C 1-10 "Alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. 1-8 "Alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. 1-6 "Alkyl" means a straight-chain or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers.

[0090] Term "C" 2-40"Alkenyl" should be understood as representing a straight-chain or branched monovalent hydrocarbon group containing one or more double bonds and having 2 to 40 carbon atoms, preferably "C". 2-10 "Alkenyl". "C" 2-10 "Alkenyl" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, more preferably "C 2-8 "Alkenyl". "C" 2-10 "Alkenyl" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7 or 8 carbon atoms, for example, having 2, 3, 4, 5 or 6 carbon atoms (i.e., C... 2-6 alkenyl), having 2 or 3 carbon atoms (i.e., C24, C34, C4 ... 2-3 Alkenyl). It should be understood that when the alkenyl group contains more than one double bond, the double bonds may be separable or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)- Pentyl-1-enyl, (Z)-pentyl-1-enyl, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl 2-Methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methyl But-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0091] Term "C" 2-40 "Alkyne group" should be understood as representing a straight or branched monovalent hydrocarbon group containing one or more triple bonds and having 2 to 40 carbon atoms, preferably "C". 2-10 "Alkyne group". The term "C"2-10 "Alkyne" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, for example, having 2, 3, 4, 5, 6, 7, or 8 carbon atoms (i.e., "C"). 2-8 "Alkyne group" has 2, 3, 4, 5 or 6 carbon atoms (i.e., "C"). 2-6 The alkynyl group ("C") has 2 or 3 carbon atoms ("C") 2-3 The alkynyl group is, for example, ethynyl, prop-1-alkynyl, prop-2-alkynyl, but-1-alkynyl, but-2-alkynyl, but-3-alkynyl, pent-1-alkynyl, pent-2-alkynyl, pent-3-alkynyl, pent-4-alkynyl, hex-1-alkynyl, hex-2-alkynyl, hex-3-alkynyl, hex-4-alkynyl, hex-5-alkynyl, 1-methylprop-2-alkynyl, 2-methylbut-3-alkynyl, 1-methylbut-3-alkynyl, 1-methylbut-2-alkynyl, 3-methylbut-1-alkynyl, 1-ethylprop-2-alkynyl, 3-methylpent-4-alkynyl, 2-methylpent-4-alkynyl, 1-methylpent-4-alkynyl -Alynyl, 2-methylpentan-3-ynyl, 1-methylpentan-3-ynyl, 4-methylpentan-2-ynyl, 1-methylpentan-2-ynyl, 4-methylpentan-1-ynyl, 3-methylpentan-1-ynyl, 2-ethylbutan-3-ynyl, 1-ethylbutan-3-ynyl, 1-ethylbutan-2-ynyl, 1-propylpropan-2-ynyl, 1-isopropylpropan-2-ynyl, 2,2-dimethylbutan-3-ynyl, 1,1-dimethylbutan-3-ynyl, 1,1-dimethylbutan-2-ynyl, or 3,3-dimethylbutan-1-ynyl. In particular, the ynyl group is ethynyl, propan-1-ynyl, or propan-2-ynyl.

[0092] Term "C" 3-40 "Cycloalkyl" should be understood to refer to saturated monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring) hydrocarbon rings or tricyclic alkanes, having 3 to 40 carbon atoms, preferably "C". 3-10 "Cycloalkyl", more preferably "C" 3-8 cycloalkyl. The term "C" 3-10 "Cycloalkyl" should be understood to refer to a saturated monovalent monocyclic, bicyclic (e.g., bridged, spirocyclic) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The C... 3-10Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or bicyclic hydrocarbon groups, such as borneolyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl; or tricyclic hydrocarbon groups, such as adamantyl.

[0093] The term "3-20 membered heterocyclic group" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, 6-, or 7-membered monocyclic ring, a 7-, 8-, 9-, 10-, 11-, or 12-membered bicyclic ring (such as a fused ring, bridged ring, or spirocyclic ring), or a 10-, 11-, 12-, 13-, 14-, or 15-membered tricyclic ring system, and contains at least one, for example, 1, 2, 3, 4, 5, or more heteroatoms selected from O, S, and N, wherein N and S may optionally be oxidized to various oxidation states to form nitrides, -S(O)-, or -S(O)2- states. Preferably, the heterocyclic group may be selected from "3-10 membered heterocyclic groups". The term "3-10 membered heterocyclic group" means a saturated or unsaturated non-aromatic ring or ring system containing at least one heteroatom selected from O, S, and N. The heterocyclic group can be connected to the rest of the molecule via any one of the carbon atoms or a nitrogen atom (if present). The heterocyclic group can include fused or bridged rings and spirocyclic rings. Specifically, the heterocyclic group can include, but is not limited to: 4-membered rings, such as azirrobutyl or oxobutyl; 5-membered rings, such as tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoalkyl, pyrazolealkyl, or pyrrololinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazineyl, or trithiaalkyl; or 7-membered rings, such as diazacycloheptyl. Optionally, the heterocyclic group can be benzofused. The heterocyclic group can be bicyclic, such as, but not limited to, a 5,5-membered ring, like a hexahydrocyclopentano[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, like a hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The heterocyclic group can be partially unsaturated, meaning it can contain one or more double bonds, such as, but not limited to, dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrroleyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl, or 4H-[1,4]thiazinyl, or it can be benzofused, such as, but not limited to, dihydroisoquinolinyl. When the 3-20-membered heterocyclic group is linked to other groups to form the compounds of the present invention, the carbon atom on the 3-20-membered heterocyclic group can be linked to other groups, or the heterocyclic atom on the 3-20-membered heterocyclic ring can be linked to other groups. For example, when the 3-20 membered heterocyclic group is selected from piperazine, the nitrogen atom on the piperazine group can be attached to other groups. Or when the 3-20 membered heterocyclic group is selected from piperidinium, the nitrogen atom on the piperidinium ring and the carbon atom at its para position can be attached to other groups.

[0094] Term "C" 6-20 "Aryl" should preferably be understood to represent a monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring), or tricyclic hydrocarbon ring having 6 to 20 carbon atoms and possessing monovalent aromaticity or partial aromaticity. It can be a monoaromatic ring or a polyaromatic ring fused together, preferably "C". 6-14Aryl. The term "C" 6-14 "Aryl" should be understood to preferably represent a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring ("C") having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms. 6-14 Aryl), particularly a ring with 6 carbon atoms (“C6 aryl”), such as phenyl; or biphenyl, or a ring with 9 carbon atoms (“C9 aryl”), such as indenyl or indenyl, or a ring with 10 carbon atoms (“C9 aryl”). 10 Aryl groups, such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl, or rings with 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups, or rings with 14 carbon atoms (“C”). 14 Aryl), for example, anthracene. When the C 6-20 When the aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, it can be ortho, para, or meta substituted.

[0095] The term "5-20-membered heteroaryl" should be understood to include monocyclic, bicyclic (e.g., fused, bridged, spirocyclic), or tricyclic aromatic ring systems having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O, and S, such as "5-14-membered heteroaryl". The term "5-14-membered heteroaryl" should also be understood to include monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5, 6, 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms independently selected from N, O, and S, and in each case, may be benzofused. Examples of monocyclic "heteroaryl" include, for example, pyridinyl, pyrimidinyl, pyrazinyl, thiazinyl, oxazinyl, triazinyl, thiadiazinyl, or oxadiazinyl. "Heteroaryl" also refers to a group in which a heteroaryl ring is fused with one or more aryl, alicyclic, or heterocyclic rings, wherein the attachment site is on the heteroaryl ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indazinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinazinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinazinyl. 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthidyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-pyrolinyl, 2-, 4-, 6- or 7-pteridyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aH Carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbazolyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenanthridyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-acridyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-pyridyl, 2-, 3-, 4-, 5-, 6-, 8- , 9- or 10-phenanthroline, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenthiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl, 2-, 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzisoquinolinyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazino[2,3-c]carbazole, 2-, 3-, 5-, 6- or 7-2H-furano[3,2-b]-pyranyl, 2-, 3-, 4-, 5-, 7- or 8-5H-pyrido[2,3-d]-o-azinyl, 1-, 3- or 5-1H-pyridinyl Azo[4,3-d]-azole, 2-, 4- or 54H-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-furano[3,4-c]cenolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10-, or 11-4H-pyrido[2,3-c]carbazole, 2-, 3-, 6-, or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thiophene, 2-, 4-, 5-, 6-, or 7-benzozolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-, 4-, 5-, 6-, or 7-benzothiazolyl 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7- or 8-benzoazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-4H-pyrrolo[1,2-b][2]benzozapinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thiophene, 2-, 4-, 5-, 6-, or 7-benzozozolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5-20-membered heteroaryl group is linked to other groups to form the compounds of the present invention, the carbon atom on the 5-20-membered heteroaryl ring may be linked to other groups, or the heteroatom on the 5-20-membered heteroaryl ring may be linked to other groups. When the 5-20-membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution sites; for example, hydrogen atoms bonded to carbon atoms on the heteroaryl ring can be substituted, or hydrogen atoms bonded to heteroatoms on the heteroaryl ring can be substituted.

[0096] The term "spirocycle" refers to a ring system in which two rings share a single ring atom.

[0097] The term "fused ring" refers to a ring system in which two rings share two cyclic atoms.

[0098] The term "bridged ring" refers to a ring system in which two rings share three or more cyclic atoms.

[0099] Unless otherwise stated, heterocyclic, heteroaryl, or heteroaryl groups include all possible isomers, such as their positional isomers. Thus, for some illustrative, non-limiting examples, forms may include those in which one, two, or more of the following positions (if present) are substituted or bonded to other groups, including pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thiophene or thiophene groups include thiophene-2-yl, thiophene-2-yl, thiophene-3-yl, and thiophene-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0100] The term "oxo" refers to the substitution of a carbon, nitrogen, or sulfur atom in a substituent with an oxygen atom (=O) formed by oxidation.

[0101] The term "alkylamino" refers to -NH-(alkyl) or -N-(alkyl)2, where alkyl is defined as described above. Non-limiting examples of alkylamino include: methylamino, ethylamino, propylamino, isopropylamino, butylamino, dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, etc.

[0102] The term "alkyloxy" refers to -O-(alkyl), where alkyl is defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, and butoxy. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkyloxy, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, or heterocycloalkyloxy.

[0103] The terms "alkyleneoxy" and "oxyalkylene" refer to -alkylene-O- or -O-alkylene-, where alkylene represents a straight-chain or branched saturated divalent hydrocarbon group. The definition of the number of carbon atoms in "alkylene" follows the definition of "alkyl" above. Those skilled in the art will understand that alkyleneoxy or oxyalkylene can be attached to the remainder of the molecule containing it in any orientation; that is, the two are used interchangeably.

[0104] "Halogenated alkyl" refers to an alkyl group that has been substituted with one or more halogens, wherein the alkyl group is as defined above.

[0105] In this invention, the compounds involved also include isotopically labeled compounds, which are the same as those shown in Formula I, but in which one or more atoms are replaced by atoms with atomic masses or mass numbers different from those normally found in nature. Examples of isotopes that can be incorporated into the compounds of this invention include isotopes of H, C, N, O, S, F, and Cl, respectively such as 2 H, 3 H, 13 C 11 C 14 C 15 N、 18 O、 17 O、 32 P, 35 S, 18 F and 36 Cl. Compounds of the present invention, their prodrugs, or pharmaceutically acceptable salts of said compounds or prodrugs containing the aforementioned isotopes and / or other isotopes are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as those doped with radioactive isotopes (e.g.,... 3 H and 14 Compounds in (C) can be used for drug and / or substrate tissue distribution assays. Tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, heavier isotopes (such as deuterium, i.e., 2 Hydrogen (H or D) substitution can provide certain therapeutic advantages derived from greater metabolic stability (e.g., increased in vivo half-life or reduced dose requirement), and may therefore be preferred in certain circumstances. The compounds of the invention claimed in the claims are particularly defined as being substituted with deuterium or tritium. Furthermore, the presence of hydrogen in the substituents without a separate mention of the terms deuterium or tritium does not exclude deuterium or tritium, but may also include deuterium or tritium.

[0106] Those skilled in the art will understand that the compounds shown in Formula I can exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they can form acid addition salts; if these compounds have an acidic center, they can form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they can also form internal salts.

[0107] The compounds of the present invention may exist as solvates (such as hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, may be stoichiometric or non-stoichiometric.

[0108] Depending on their molecular structure, the compounds of the present invention can be chiral, and therefore may exist in various enantiomeric forms. Thus, these compounds can exist in racemic or optically active forms. The compounds of the present invention encompass isomers of each chiral carbon in the R or S configuration, or mixtures thereof, and racemates. The compounds of the present invention or their intermediates can be isolated as enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are obtained from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid in both R and S forms, 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. Chromatographic enantiomeric separation can also be advantageously performed using optically active resolving agents (e.g., dinitrobenzoylphenylglycine immobilized on silica gel, cellulose triacetate or other carbohydrate derivatives, or chiral derivatized isobutylene ester polymers). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, such as hexane / isopropanol / acetonitrile.

[0109] The corresponding stable isomers can be separated using known methods, such as extraction, filtration, or column chromatography.

[0110] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0111] The term “therapeutic effective amount” refers to the amount of an active compound or drug that researchers, veterinarians, physicians, or other clinicians are searching for in tissues, systems, animals, individuals, or humans to elicit a biological or medical response. It includes one or more of the following: (1) prevention of disease: e.g., prevention of disease, disorder, or condition in individuals susceptible to disease, disorder, or symptom but not yet experiencing or exhibiting the pathology or symptoms of the disease; (2) suppression of disease: e.g., suppression of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptoms of the disease, disorder, or symptom (i.e., prevention of further development of the pathology and / or symptoms); (3) relief of disease: e.g., relief of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptoms of the disease, disorder, or symptom (i.e., reversal of the pathology and / or symptoms). Detailed Implementation

[0112] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0113] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0114] Preparation of some raw materials:

[0115] 1. Synthesis of 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine

[0116]

[0117] 2,4-Dichloro-5-trifluoromethylpyrimidine (20 g, 0.089 mol) and methanol (45 mL) were added to a reaction flask. The system was stirred at -10 °C for 10 minutes. Then, triethylamine (12.5 mL, 0.089 mol) and a methanol solution of methylamine (2 M, 45 mL) were added. The system was stirred for another hour, then naturally heated and stirred overnight at room temperature. The mixture was concentrated, and water was added to the reaction system. The mixture was extracted three times with ethyl acetate. The organic phases were combined and washed with saturated brine. The solution was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography to give 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidine-4-amine (8.5 g, 45%).

[0118] MS m / z(ESI): 212[M+1] + .

[0119] 2. Synthesis of 2-chloro-N-ethyl-5-(trifluoromethyl)pyrimidin-4-amine

[0120]

[0121] For synthesis, see 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine.

[0122] MS m / z(ESI): 226[M+1] + .

[0123] 3. Synthesis of 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidine-4-amine

[0124]

[0125] For synthesis, see 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine.

[0126] MS m / z(ESI): 238[M+1] + .

[0127] 4. Synthesis of 2-chloro-N-(2,2-difluoroethyl)-5-(trifluoromethyl)pyrimidine-4-amine

[0128]

[0129] For synthesis, see 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine.

[0130] MS m / z(ESI): 262[M+1] + .

[0131] 5. Synthesis of 2-chloro-N-(2,2,2-trifluoroethyl)-5-(trifluoromethyl)pyrimidine-4-amine

[0132]

[0133] For synthesis, see 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine.

[0134] MS m / z(ESI): 280[M+1] + .

[0135] Example 1

[0136] (S)-2-amino-3-(3-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0137]

[0138] first step

[0139] Preparation of (R)-2-((benzyloxycarbonyl)amino)-3-iodopropionate benzyl ester

[0140] Triphenylphosphine (1.2 g, 4.5 mmol) and imidazole (0.3 g, 4.5 mmol) were added to a round-bottom flask, and nitrogen was purged three times. Then, dichloromethane (10 mL) and iodine (1.1 g, 4.5 mmol) were added. The reaction mixture was stirred at room temperature for 10 minutes and then cooled to 0 °C. A dichloromethane solution of (N-benzyloxycarbonyl)-L-serine benzyl ester Cpd-01A (1.0 g, 3.0 mmol) was then slowly added dropwise. The reaction mixture was stirred at 0 °C for 2 hours. After the reaction was complete, the solid in the reaction mixture was removed by filtration. The filtrate was evaporated to dryness and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1) to obtain (R)-2-((benzyloxycarbonyl)amino)-3-iodopropionate benzyl ester Cpd-01B (1.2 g, white solid), yield: 83%.

[0141] MS m / z(ESI): 462 [M+23] + .

[0142] Step 2

[0143] Preparation of (R)-(3-(benzyloxy)-2-(benzyloxycarbonyl)amino)-3-oxopropyl)zinc iodide

[0144] Activated zinc powder (0.75 g, 11.5 mmol) and iodine (0.15 g, 0.6 mmol) were added to a three-necked flask. After purging with nitrogen three times, the iodine was sublimated using a blower. Then, a solution of (R)-2-((benzyloxycarbonyl)amino)-3-iodopropionate Cpd-01B (1.0 g, 2.3 mmol) in N,N-dimethylformamide (10 mL) was added to the flask all at once. The mixture was stirred for 10 minutes while maintaining the temperature inside the flask. The reaction solution was withdrawn with a syringe and used directly in the next reaction without any further processing.

[0145] Step 3

[0146] Preparation of (S)-2-((benzyloxycarbonyl)amino)-3-(3-methoxy-4-nitrophenyl)propionate benzyl ester

[0147] 4-Bromo-2-methoxy-1-nitrobenzene (0.27 g, 1.2 mmol), bis(triphenylphosphine)palladium dichloride (0.08 g, 0.1 mmol), cuprous iodide (0.05 g, 0.3 mmol), and N,N-dimethylformamide were added to a three-necked flask. After purging with nitrogen three times, the flask was preheated in an oil bath at 80 °C for 10 minutes. Subsequently, a solution of (R)-(3-(benzyloxy)-2-(benzyloxycarbonyl)amino)-3-oxopropyl)zinc iodide Cpd-01C in N,N-dimethylformamide was slowly added dropwise using a syringe. The reaction mixture was stirred at 80 °C for 0.5 hours. After the reaction was complete, the reaction mixture was filtered through diatomaceous earth, then poured into water (200 mL), and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain (S)-2-((benzyloxycarbonyl)amino)-3-(3-methoxy-4-nitrophenyl)propionate Cpd-01D (0.45 g, brown oil), yield: 70%.

[0148] MS m / z(ESI): 487 [M+23] + .

[0149] Step 4

[0150] Preparation of (S)-3-(4-amino-3-methoxyphenyl)-2-((benzyloxycarbonyl)amino)propionate benzyl ester

[0151] (S)-2-((benzyloxycarbonyl)amino)-3-(3-methoxy-4-nitrophenyl)propionate Cpd-01D (400 mg, 0.86 mmol), iron powder (385 mg, 6.89 mmol), and ethanol (5 mL) were added to a reaction flask, followed by the addition of saturated ammonium chloride solution (5 mL). The reaction mixture was stirred at 90 °C for 1 hour. After the reaction was complete, the reaction mixture was filtered through diatomaceous earth, diluted with water, and extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by preparative agar (petroleum ether / ethyl acetate = 3 / 1) to obtain (S)-3-(4-amino-3-methoxyphenyl)-2-((benzyloxycarbonyl)amino)propionate Cpd-01E (340 mg, light brown oil), yield: 73%.

[0152] MS m / z(ESI): 435[M+1] + .

[0153] 1 H NMR(400MHz,DMSO-d6)δ7.78(d,J=7.9Hz,1H),7.50-7.09(m,10H),6.70(s,1H),6.52(s,2H) ,5.09(s,2H),4.99(s,2H),4.57(s,2H),4.33-4.16(m,1H),3.68(s,3H),2.96-2.67(m,2H).

[0154] Step 5

[0155] Preparation of (S)-2-((benzyloxycarbonyl)amino)-3-(3-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)benzyl propionate

[0156] (S)-3-(4-amino-3-methoxyphenyl)-2-((benzyloxycarbonyl)amino)propionate Cpd-01E (226 mg, 0.52 mmol) and 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (111 mg, 0.52 mmol) were added to tert-butanol (5 mL), followed by the addition of trifluoroacetic acid (5 mg, 0.052 mmol) and purging with nitrogen three times. The reaction solution was reacted at 110 °C for 1 hour. After the reaction was complete, the solvent was removed by rotary evaporation, and the residue was dispersed in ethyl acetate (5 mL). The residue was filtered, and the solid was washed twice with ethyl acetate (2 mL). After vacuum drying, benzyl (S)-2-((benzyloxycarbonyl)amino)-3-(3-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate Cpd-01F (260 mg, white solid) was obtained, with a yield of 74%.

[0157] MS m / z(ESI): 610[M+1] + .

[0158] Step 6

[0159] Preparation of (S)-2-amino-3-(3-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0160] (S)-2-((benzyloxycarbonyl)amino)-3-(3-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate Cpd-01F (250 mg, 0.41 mmol) and palladium on carbon (10 mg) were added to methanol (5 mL). The reaction mixture was stirred at room temperature for 1 hour under a hydrogen atmosphere. After the reaction was completed, the palladium on carbon was removed by diatomaceous earth filtration, and the filtrate was evaporated to dryness. The crude product was purified by reversed-phase chromatography to obtain (S)-2-amino-3-(3-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate Cpd-01 (40 mg), with a yield of 25%.

[0161] MS m / z(ESI): 386[M+1] + .

[0162] 1H NMR (400MHz, DMSO-d6) δ8.26-8.18(m,1H),8.14(s,1H),8.10(d,J=8.1Hz,1H),7.95(s,1H),7.15(d,J=4.2Hz,1H),6.9 8(s,1H),6.82(d,J=8.2Hz,1H),3.85(s,3H),3.50(s,1H),3.22-3.06(m,1H),2.90(d,J=4.4Hz,3H),2.87-2.77(m,1H).

[0163] Example 2

[0164] (S)-2-amino-3-(4-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid

[0165]

[0166] Following the synthetic steps of Example 1, except that 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine was replaced with 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine, yielding compound (S)-2-amino-3-(4-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-02 (180 mg), yield 62%. MS m / z (ESI): 400 [M+1] + .

[0167] 1 H NMR (400MHz, DMSO-d6) δ8.14(s,1H),8.05(d,J=8.2Hz,1H),7.94(s,1H),7.38(s,2H),7.15(s,1H),6.97(s,1H),6.81(d,J=8.3Hz,1H ),3.84(d,J=7.9Hz,3H),3.46(m,2H),3.38(m,1H),3.14(dd,J=14.3,3.9Hz,1H),2.79(dd,J=14.4,8.8Hz,1H),1.15(t,J=7.1Hz,3H).

[0168] Example 3

[0169] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid

[0170]

[0171] The synthesis steps of Example 1 were followed, except that 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine was replaced with 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidin-4-amine to obtain (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-03 (45 mg), yield: 30%. MS m / z (ESI): 412 [M+1] + .

[0172] 1 H NMR (400MHz, DMSO-d6) δ8.34(s,1H),8.17(s,1H),7.95(s,1H),7.29(d,J=73.0Hz,3H),6.98(s,1H),6.82(d,J=8.3Hz,1H),3.86(s ,3H),3.45-3.24(m,1H),3.14(dd,J=14.4,3.6Hz,1H),2.79(dd,J=14.4,8.8Hz,2H),0.78(d,J=5.4Hz,2H),0.67(d,J=2.8Hz,2H).

[0173] Example 4

[0174] (S)-2-amino-3-(4-((4-((2,2-difluoroethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid

[0175]

[0176] Following the synthetic steps of Example 1, except that 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine was replaced with 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine, to obtain compound (S)-2-amino-3-(4-((4-(((2,2-difluoroethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-04 (50 mg), yield: 33%.

[0177] MS m / z(ESI): 436[M+1] + .

[0178] 1H NMR (400MHz, DMSO-d6) δ8.30(s,1H),8.21(s,1H),7.82(d,J=8.1Hz,1H),7.37(s,3H),6.99(s,1H),6.80(d,J=8.1Hz,1H),6.20(t,J=56.6Hz ,1H),3.83(s,3H),3.76(t,J=12.6Hz,2H),3.36(ddt,J=36.7,26.0,5.2Hz,1H),3.14(dd,J=14.3,3.7Hz,1H),2.79(dd,J=14.2,8.6Hz,1H).

[0179] Example 5

[0180] Preparation of (S)-2-amino-3-(3-methoxy-4-((4-((2,2,2-trifluoroethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0181]

[0182] Following the synthetic steps of Example 1, except that 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine was replaced with 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine, to obtain compound (S)-2-amino-3-(3-methoxy-4-((4-((2,2,2-trifluoroethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid Cpd-05 (100 mg), yield: 57%.

[0183] MS m / z(ESI): 454[M+1] + .

[0184] 1 H NMR (400MHz, DMSO-d6) δ8.30(s,1H),8.26(s,1H),7.82(d,J=8.1Hz,1H),7.54(br,3H),6.99(d,J=1.2Hz,1H),6.87–6.73(m,1H ), 4.23 (d, J = 9.3Hz, 2H), 3.82 (s, 3H), 3.38 (dd, J = 8.6, 4.1Hz, 1H), 3.14 (dd, J = 14.6, 4.2Hz, 1H), 2.80 (dd, J = 14.3, 8.7Hz, 1H).

[0185] Example 6

[0186] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-N-methylpropionamide

[0187]

[0188] Steps one through four were performed sequentially as steps two through five in Example 1, with the difference that compound Cpd-07A was used as the starting compound to replace (R)-2-((benzyloxycarbonyl)amino)-3-iodopropionate benzyl ester Cpd-01B and 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidin-4-amine was used to replace 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine, to obtain compound (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate methyl ester Cpd-07E (1.8 g, white solid), yield: 35%.

[0189] MS m / z(ESI): 526 [M+1] + Step 5

[0190] Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid

[0191] Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate Cpd-07E (1.8 g, 3.4 mmol) was dissolved in tetrahydrofuran (10 mL), and an aqueous solution of lithium hydroxide (0.4 g, 10.2 mmol) (10 mL) was added dropwise. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the pH was adjusted to 4 with 1 mol / L HCl. The precipitated solid was collected by filtration and washed with methanol and ethyl acetate. After vacuum drying, (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate Cpd-07F (1.2 g, white solid) was obtained, yield: 65%.

[0192] MS m / z(ESI): 512[M+1] + .

[0193] Step 6

[0194] Preparation of (S)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-1-(methylamino)-1-oxopropane-2-yl)carbamate tert-butyl

[0195] (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-07F (100 mg, 19.5 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (148 mg, 39.0 mmol), and N,N-diisopropylethylamine (126 mg, 97.5 mmol) were added to N,N-dimethylformamide (10 mL). After reacting at room temperature for 30 minutes, methylamine hydrochloride (20 mg, 29.2 mmol) was added, and the reaction was continued for 2 hours. After the reaction was completed, water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed three times with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain (S)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-1-(methylamino)-1-oxopropane-2-yl)carbamate tert-butyl Cpd-07G (100 mg, white solid), yield: 92%.

[0196] MS m / z(ESI): 525 [M+1] + .

[0197] Step 7

[0198] Preparation of (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-N-methylpropylamine

[0199] (S)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-1-(methylamino)-1-oxopropane-2-yl)carbamate tert-butyl ester Cpd-07G (100 mg, 19.0 mmol) was dissolved in 10 mL of dioxane HCl and stirred at room temperature for 1 hour. The reaction solution was concentrated and dried under vacuum to give (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-N-methylpropionamide Cpd-07 (70 mg) hydrochloride, yield: 82%.

[0200] MS m / z(ESI): 425[M+1]+ .

[0201] 1 H NMR(400MHz,MeOD)δ8.26(s,2H),7.05(s,1H),6.92(d,J=8.0Hz,1H),4.13–4.00(m,1H),3.98(s,3H),3.20(dd,J =13.8,6.9Hz,1H),3.08(dd,J=13.6,7.6Hz,1H),2.96(s,1H),2.74(s,3H),1.04–0.86(m,2H),0.86–0.71(m,2H).

[0202] Example 7

[0203] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-N,N-dimethylpropionamide

[0204]

[0205] Step 1: Following the synthesis steps of Example 6, compound Cpd-07F was obtained. Then, following step 6 of Example 6, methylamine hydrochloride was replaced with dimethylamine hydrochloride.

[0206] Step 2: Referring to Step 7 of Example 6, the hydrochloride salt of compound (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-N,N-dimethylpropionamide Cpd-08 (70 mg, white solid) was obtained with a yield of 90%.

[0207] MS m / z(ESI): 439[M+1] + .

[0208] 1 H NMR(400MHz,MeOD)δ8.29(s,2H),7.05(s,1H),6.95(d,J=8.2Hz,1H),4.69(t,J=7.3Hz,1H),3.98(s,3H), 3.14(d,J=7.3Hz,2H),3.03–2.94(m,1H),2.93(s,3H),2.82(s,3H),0.99–0.90(m,2H),0.86–0.75(m,2H).

[0209] Example 8

[0210] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate isopropyl ester

[0211]

[0212] first step

[0213] Preparation of isopropyl (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate

[0214] Isopropanol (40.3 mg, 67.1 mmol) was cooled to 0 °C, followed by the slow dropwise addition of acetyl chloride (5.8 mg, 10.9 mmol). The mixture was stirred at 0 °C for 1 hour, and then (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-07F (80.0 mg, 15.6 mmol) was added. The mixture was then heated to 70 °C and reacted overnight. After the reaction was complete, the reaction solution was concentrated and dried under vacuum to give isopropyl (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid isopropyl ester Cpd-09 (50 mg) hydrochloride, yield: 66%.

[0215] MS m / z(ESI): 454[M+1] + .

[0216] 1 H NMR(400MHz,MeOD)δ8.36(s,1H),8.24(s,1H),7.04(s,1H),6.93(d,J=8.3Hz,1H),5.15–4.99(m,1H),4.31(t,J=7.0Hz,1H),3.97 (s,3H),3.28–3.13(m,2H),3.00–2.86(m,1H),1.29(d,J=6.2Hz,3H),1.23(d,J=6.2Hz,3H),0.97–0.88(m,2H),0.81–0.72(m,2H).

[0217] Example 9

[0218] (S)-2-amino-3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0219]

[0220] Following the synthetic steps of Example 1, compound Cpd-01C was first prepared. Then, in step three, 4-bromo-2-methoxy-1-nitrobenzene was replaced with 1-bromo-2-fluoro-5-methoxy-4-nitrobenzene, and the reaction continued, ultimately yielding compound (S)-2-amino-3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid Cpd-10 (180 mg), yield 72%. MS m / z (ESI): 404 [M+1] + .

[0221] 1 H NMR (400MHz, DMSO-d6) δ8.39(br,2H),8.25-8.16(m,2H),8.03(s,1H),7.32(d,J=4.3Hz,1H),7.06(d,J=7 .0Hz,1H),4.08(t,J=6.6Hz,1H),3.87(s,3H),3.12(ddd,J=35.9,14.3,6.7Hz,2H),2.93(d,J=4.3Hz,3H).

[0222] Example 10

[0223] (S)-2-amino-3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate isopropyl ester

[0224]

[0225] Following the synthetic steps of Example 8, starting with compound Cpd-10, isopropyl (S)-2-amino-3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate isopropyl hydrochloride Cpd-11 (27 mg) was obtained in 75% yield. MS m / z (ESI): 446 [M+1] + .

[0226] 1 H NMR(400MHz,MeOD)δ8.32(s,1H),8.05(s,1H),7.10(d,J=6.7Hz,1H),5.06(dt,J=12.5,6 .3Hz,1H),4.28(t,J=7.2Hz,1H),3.97(s,3H),3.14(s,3H),1.23(dd,J=23.6,6.3Hz,6H).

[0227] Example 11

[0228] 4-Amino-3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)butyric acid

[0229]

[0230] first step

[0231] Preparation of tert-butyl (4-bromo-5-fluoro-2-methoxyphenyl)carbamate

[0232] 4-Bromo-5-fluoro-2-methoxyaniline Cpd-12A (2.0 g, 9.2 mmol), di-tert-butyl dicarbonate (3.0 g, 6.5 mmol), and cesium carbonate (9.9 g, 45.8 mmol) were dissolved in methanol (60 mL) and stirred overnight at 70 °C. After the reaction was complete, the crude product was obtained by rotary evaporation and purified by column chromatography (petroleum ether / ethyl acetate = 20 / 1) to give (4-bromo-5-fluoro-2-methoxyphenyl)carbamate tert-butyl Cpd-12B (2.25 g, pale yellow oil), yield: 77%.

[0233] MS m / z(ESI): 320[M+1] + .

[0234] Step 2

[0235] Preparation of (E)-3-(4-((tert-butoxycarbonyl)amino)-2-fluoro-5-methoxyphenyl)acrylate

[0236] (4-Bromo-5-fluoro-2-methoxyphenyl)carbamate tert-butyl ester Cpd-12B (500 mg, 1.57 mmol), (E)-2-(ethoxycarbonyl)vinylboronic acid pinacol ester (530 mg, 2.35 mmol), bis(triphenylphosphine)palladium dichloride (110 mg, 0.16 mmol), and sodium carbonate (248 mg, 2.35 mmol) were dissolved in 1,4-dioxane (10 mL) and water (1 mL) and reacted at 100 °C for 12 hours. After the reaction was completed, the mixture was cooled to room temperature and filtered through diatomaceous earth. The reaction solution was quenched with water (5 mL), extracted with ethyl acetate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 9 / 1) to give (E)-3-(4-((tert-butoxycarbonyl)amino)-2-fluoro-5-methoxyphenyl)acrylate Cpd-12C (450 mg, white oil), yield: 84.5%.

[0237] MS m / z(ESI): 340[M+1] + .

[0238] Step 3

[0239] Preparation of ethyl 3-(4-((tert-butoxycarbonyl)amino)-2-fluoro-5-methoxyphenyl)-4-nitrobutyrate

[0240] Nitromethane (0.8 g, 13.2 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.0 g, 6.6 mmol) were dissolved in acetonitrile (7.5 mL) and stirred at 0 °C for 10 minutes. Ethyl (E)-3-(4-((tert-butoxycarbonyl)amino)-2-fluoro-5-methoxyphenyl)acrylate Cpd-12C (450 mg, 1.3 mmol) was slowly added to the above reaction solution, and the reaction was carried out at 0 °C for 12 hours. After the reaction was completed, the reaction solution was poured into a large amount of water, the pH was adjusted to 2 with 10% HCl, and the product was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give ethyl 3-(4-((tert-butoxycarbonyl)amino)-2-fluoro-5-methoxyphenyl)-4-nitrobutyrate Cpd-12D (0.39 g, white oil), yield: 73.8%.

[0241] MS m / z(ESI): 401[M+1] + .

[0242] 1 H NMR (400MHz, CDCl3) δ7.93(d,J=12.6Hz,1H),7.74(d,J=16.1Hz,1H),7.22(s,1H),6.88(d,J=6.4Hz,1H),6.36(d,J =16.1Hz,1H),4.23(q,J=7.1Hz,2H),3.86(s,3H),1.51(s,9H),1.31(t,J=7.1Hz,3H),0.85(dt,J=8.3,3.0Hz,2H).

[0243] Step 4

[0244] Preparation of ethyl 3-(4-amino-2-fluoro-5-methoxyphenyl)-4-nitrobutyrate

[0245] Ethyl 3-(4-((tert-butoxycarbonyl)amino)-2-fluoro-5-methoxyphenyl)-4-nitrobutyrate Cpd-12D (0.39 g, 0.98 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2.2 g, 19.5 mmol) was added. The mixture was stirred at 25 °C for 1 hour. After the reaction was complete, rotary evaporation yielded ethyl 3-(4-amino-2-fluoro-5-methoxyphenyl)-4-nitrobutyrate Cpd-12E (0.4 g, white solid), yield: 99%.

[0246] MS m / z(ESI): 301[M+1] + .

[0247] Step 5

[0248] Preparation of ethyl 3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-4-nitrobutyrate

[0249] Ethyl 3-(4-amino-2-fluoro-5-methoxyphenyl)-4-nitrobutyrate Cpd-12E (0.4 g, 0.96 mmol) and 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (0.2 g, 0.98 mmol) were dissolved in n-butanol (10 mL), and trifluoroacetic acid (0.12 g, 0.98 mmol) was added. The mixture was reacted at 90 °C for 1 hour. After the reaction was completed, the mixture was cooled to room temperature, quenched with water (5 mL), extracted with ethyl acetate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (dichloromethane / methanol = 30 / 1) to give ethyl 3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-4-nitrobutyrate Cpd-12F (0.21 g, white solid), yield: 46.8%.

[0250] MS m / z(ESI): 476[M+1] + .

[0251] Step 6

[0252] Preparation of 3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-4-nitrobutyric acid

[0253] Ethyl 3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-4-nitrobutyric acid Cpd-12F (0.12 g, 0.25 mmol) was dissolved in tetrahydrofuran (5 mL), and saturated lithium hydroxide aqueous solution (0.5 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the pH was adjusted to 4 with dilute hydrochloric acid, and the mixture was extracted with a mixed solvent (dichloromethanol / methanol = 10 / 1). The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography to obtain 3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-4-nitrobutyric acid Cpd-12G (0.1 g, white solid), yield: 89%.

[0254] MS m / z(ESI): 448[M+1] + .

[0255] Step 7

[0256] Preparation of 4-amino-3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)butyric acid

[0257] 0.1 g (0.22 mmol) of 3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-4-nitrobutyric acid Cpd-12G was dissolved in methanol (5 mL), and Raney nickel (0.5 mL) was added. The reaction was carried out at room temperature under a hydrogen atmosphere for 2 hours. After the reaction was completed, the mixture was filtered through diatomaceous earth and rotary evaporated to obtain 0.02 g (4-amino-3-(2-fluoro-5-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)butyric acid Cpd-12, yield: 21.8%.

[0258] MS m / z(ESI): 418[M+1] + .

[0259] 1 H NMR (400MHz, DMSO-d6) δ8.19(s,1H),8.09(d,J=12.3Hz,1H),7.96(s,1H),7.25(s,1H),6.92(d,J=6.6Hz ,1H),3.84(s,3H),3.30(s,9H),2.92(d,J=3.7Hz,3H),2.80(s,1H),2.25(d,J=11.7Hz,1H),1.23(s,2H).

[0260] Example 12

[0261] 4-Amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)butyric acid

[0262]

[0263] Following the synthetic steps of Example 11, except that in the first step of Example 11, 4-bromo-2-methoxyaniline Cpd-13A was replaced with 4-bromo-5-fluoro-2-methoxyaniline Cpd-12A, and in the fifth step, 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidin-4-amine was replaced with 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine, to obtain compound 4-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)butyric acid Cpd-13 (68 mg), yield: 29%. MS m / z (ESI): 426 [M+1] + .

[0264] 1 H NMR(400MHz,DMSO-d6)δ8.34(s,1H),8.17(s,1H),7.97(s,1H),7.19(s,1H),6.9 3(s,1H),6.80(d,J=8.3Hz,1H),3.87(s,3H),3.06(dd,J=10.2,5.7Hz,1H),2.99( d,J=8.9Hz,1H),2.90(dd,J=11.7,3.4Hz,1H),2.82(d,J=3.1Hz,1H),2.71(dd,J =16.1,9.0Hz,1H),2.42(d,J=13.0Hz,1H),0.84–0.70(m,2H),0.70–0.50(m,2H).

[0265] Example 13

[0266] (S)-2-amino-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionic acid

[0267]

[0268] first step

[0269] Preparation of 1-bromo-2-chloro-5-methoxy-4-nitrobenzene

[0270] 1-Bromo-2-chloro-5-fluoro-4-nitrobenzene Cpd-16A (2.0 g, 7.9 mmol) was dissolved in methanol (20 mL), stirred at 0 °C, and sodium methoxide (1.4 g, 7.9 mmol) was added. The mixture was reacted at 0 °C for 1.5 h. After the reaction was complete, water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 9 / 1) to give 1-bromo-2-chloro-5-methoxy-4-nitrobenzene Cpd-16B (2.1 g, yellow solid), yield: 95%.

[0271] 1 H NMR (400MHz, CDCl3) δ7.99(s,1H),7.36(s,1H),3.97(s,3H).

[0272] Step 2

[0273] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-5-methoxy-4-nitrophenyl)propionate

[0274] Add zinc powder (0.98 g, 18.8 mmol) and iodine (0.19 g, 0.91 mmol) to a three-necked flask, purging with nitrogen, and heat until the iodine sublimates. Dissolve 1-bromo-2-chloro-5-methoxy-4-nitrobenzene Cpd-16B (1.0 g, 3.70 mmol) in N,N-dimethylformamide (10 mL), and quickly pour the solution into a three-necked flask with stirring. Simultaneously, add cuprous iodide (0.07 g, 0.40 mmol), palladium dichloride bis(triphenylphosphine) dichloride (0.11 g, 0.20 mmol), methyl (R)-2-((tert-butoxycarbonyl)amino)-3-iodopropionate (0.40 g, 1.80 mmol), and N,N-dimethylformamide (10 mL) to another three-necked flask, purging with nitrogen, and preheat to 80 °C. A mixed solution containing zinc powder was drawn up with a syringe and slowly dripped into a three-necked flask reacting at 80°C. The reaction was continued for 1 hour. After the reaction was completed, the solution was cooled to room temperature and filtered through diatomaceous earth. The solution was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-5-methoxy-4-nitrophenyl)propionate Cpd-16C (400 mg, yellow oil), yield: 28%.

[0275] MS m / z(ESI): 389[M+1] + .

[0276] Step 3

[0277] Preparation of methyl (S)-3-(4-amino-2-chloro-5-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propionate

[0278] Methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-5-methoxy-4-nitrophenyl)propionate Cpd-16C (400 mg, 1.03 mmol), iron powder (287 mg, 5.15 mmol), and saturated aqueous solution of ammonium chloride (275 mg, 5.15 mmol) were dissolved in ethanol (5 mL) and refluxed for 2 hours. After the reaction was completed, the mixture was cooled to room temperature and filtered through diatomaceous earth. The reaction solution was quenched with water (5 mL), extracted with ethyl acetate (10 mL × 3), washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain methyl (S)-3-(4-amino-2-chloro-5-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propionate Cpd-16D (200 mg, yellow oil), yield: 54%.

[0279] MS m / z(ESI): 359[M+1] + .

[0280] Step 4

[0281] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionate

[0282] Methyl (S)-3-(4-amino-2-chloro-5-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propionate Cpd-16D (200 mg, 0.56 mol), 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidin-4-amine (160 mg, 0.67 mmol) and trifluoroacetic acid (32 mg, 0.28 mmol) were dissolved in n-butanol (10 mL) and reacted at 90 °C for 1 hour. After the reaction was completed, the mixture was cooled to room temperature, quenched with water (5 mL), extracted with ethyl acetate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionate Cpd-16E (100 mg, white solid), yield: 32%.

[0283] MS m / z(ESI): 560[M+1] + .

[0284] Step 5

[0285] Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionic acid

[0286] Methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionate Cpd-16E (100 mg, 0.18 mmol) was dissolved in tetrahydrofuran (5 mL), and lithium hydroxide aqueous solution (0.5 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the pH was adjusted to 4 with dilute hydrochloric acid, and the mixture was extracted with a mixed solvent (dichloromethanol / methanol = 10 / 1). The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionate Cpd-16F (70 mg, white solid), yield: 87%.

[0287] MS m / z(ESI): 546 [M+1] + .

[0288] Step 6

[0289] Preparation of (S)-2-amino-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionic acid

[0290] (S)-2-((tert-butoxycarbonyl)amino)-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionic acid Cpd-16F (70 mg, 0.13 mmol) was added to a hydrogen chloride methanol solution (4 mL, 4 mol / L). The reaction solution was reacted at 25 °C for 4 hours. After the reaction was completed, the reaction solution was concentrated to give (S)-2-amino-3-(2-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionic acid Cpd-16 (30 mg) hydrochloride, yield: 51%.

[0291] MS m / z(ESI): 446[M+1] + .

[0292] 1H NMR(400MHz,DMSO-d6)δ8.73(s,1H),8.23(s,1H),8.02(s,1H),7.39(s,1H),7.10(s,1H),3.88(s, 3H), 3.61 (s, 1H), 3.37 (d, J = 5.1Hz, 1H), 2.91-2.70 (m, 2H), 0.93-0.85 (m, 2H), 0.74-0.66 (m, 2H).

[0293] Example 14

[0294] (S)-2-amino-3-(5-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypyridin-2-yl)propionic acid

[0295]

[0296] Referring to the synthesis steps of Example 13, except that compound Cpd-16A was replaced with compound 2,4-dibromo-5-nitropyridine Cpd-17A to obtain compound (S)-2-amino-3-(5-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypyridin-2-yl)propionic acid Cpd-17.

[0297] MS m / z(ESI): 413[M+1] + .

[0298] 1 H NMR (400MHz, DMSO-d6) δ9.40(s,1H),8.88(s,1H),8.27(s,1H),7.47(d,J=18.4Hz,2H),4.47(t,J=6.7Hz,1H),4.04(s,3H),3 .44(dd,J=15.3,6.3Hz,1H),3.33(dd,J=15.3,7.2Hz,1H),2.97–2.72(m,1H),0.79(dd,J=6.9,4.6Hz,2H),0.71–0.60(m,2H).

[0299] Example 15

[0300] (S)-2-amino-3-(2-cyano-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionic acid

[0301]

[0302] first step

[0303] Preparation of 2-bromo-4-fluoro-5-nitrobenzene

[0304] 2-Bromo-4-fluorobenzonitrile Cpd-18A (3.0 g, 15 mmol) and concentrated sulfuric acid (5 mL) were added to a reaction flask, and potassium nitrate (1.6 g, 16 mmol) was added at 0 °C. The reaction was carried out at room temperature for 1.5 hours. After the reaction was completed, the mixture was poured into ice water (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and concentrated to give 2-bromo-4-fluoro-5-nitrobenzene Cpd-18B (3.4 g, pale yellow solid), yield: 83%.

[0305] 1 H NMR (400MHz, DMSO-d6) δ8.89 (d, J = 7.6 Hz, 1H), 8.38 (d, J = 10.7 Hz, 1H).

[0306] Referring to the synthesis steps of Example 13, the difference is that in the first step of Example 13, compound Cpd-16A was replaced with compound Cpd-18B to obtain (S)-2-amino-3-(2-cyano-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-5-methoxyphenyl)propionic acid Cpd-18.

[0307] MS m / z(ESI): 437[M+1] + .

[0308] 1 H NMR (400MHz, DMSO-d6) δ13.90(s,1H),9.32(s,1H),8.86(s,1H),8.66(s,3H),8.48(s,1H),8.19(s,1H),7.45( s,1H),4.25(d,J=4.8Hz,1H),3.98(s,3H),3.47–3.22(m,2H),2.84(dt,J=6.9,3.1Hz,1H),1.02–0.67(m,4H).

[0309] Example 16

[0310] (S)-2-amino-3-(4-methoxy-3-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0311]

[0312] first step

[0313] Preparation of (S)-2-((benzyloxycarbonyl)amino)-3-(4-hydroxy-3-nitrophenyl)propionate benzyl ester

[0314] (Benzyloxycarbonyl)-L-tyrosine benzyl ester Cpd-21A (1.0 g, 2.5 mmol) was dissolved in dichloromethane (10 mL), and 20% dilute nitric acid (240 mg, 3.7 mmol) was slowly added. The reaction mixture was allowed to react at room temperature for 8 hours. After the reaction was complete, the mixture was diluted with dichloromethane (20 mL) and washed with water (50 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give (S)-2-((benzyloxycarbonyl)amino)-3-(4-hydroxy-3-nitrophenyl)propionate benzyl ester Cpd-21B (730 mg, yellow solid), yield: 60%.

[0315] 1 H NMR (400MHz, DMSO-d6) δ10.83(s,1H),7.90(d,J=8.2Hz,1H),7.80(d,J=1.8Hz,1H),7.43(dd,J=8.5,2.1Hz,1H),7.30(tt,J=7.6,6.6Hz,10H ),7.02(d,J=8.5Hz,1H),5.12(s,2H),4.99(s,2H),4.34(td,J=9.9,5.3Hz,1H),3.05(dd,J=13.9,5.2Hz,1H),2.87(dd,J=13.8,10.3Hz,1H).

[0316] Step 2

[0317] Preparation of (S)-2-((benzyloxycarbonyl)amino)-3-(4-methoxy-3-nitrophenyl)propionate benzyl ester

[0318] (S)-2-((benzyloxycarbonyl)amino)-3-(4-hydroxy-3-nitrophenyl)propionate Cpd-21B (470 mg, 1.0 mmol) was dissolved in N,N-dimethylformamide (8 mL), and sodium bicarbonate (132 mg, 1.5 mmol) and methyl iodoforme (444 mg, 3.0 mmol) were added. The reaction mixture was reacted at 25 °C for 8 hours. After the reaction was complete, water (10 mL) was added, followed by extraction with ethyl acetate (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give (S)-2-((benzyloxycarbonyl)amino)-3-(4-methoxy-3-nitrophenyl)propionate Cpd-21C (510 mg, yellow oil), yield: 81%.

[0319] MS m / z(ESI): 487 [M+23] + .

[0320] Following the synthetic steps of Example 1, except that compound Cpd-01D was replaced with compound Cpd-21C, compound (S)-2-amino-3-(4-methoxy-3-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid Cpd-21 (230 mg) was obtained with a yield of 88%.

[0321] MS m / z(ESI): 386[M+1] + .

[0322] 1 H NMR (400MHz, DMSO-d6) δ13.83(s,1H),8.21(s,1H),8.17(s,1H),8.00(s,1H),7.21(d,J=4.4Hz,1H),7.01(d,J= 8.4Hz, 1H), 6.92 (dd, J = 8.4, 1.8Hz, 1H), 4.04 (s, 1H), 3.85 (s, 3H), 3.04 (d, J = 6.3Hz, 2H), 2.93 (d, J = 4.3Hz, 3H).

[0323] Example 17

[0324] (S)-2-amino-3-(3-methoxy-4-((4-m-tolyl-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0325]

[0326] first step

[0327] Preparation of 2-chloro-4-m-tolyl-5-(trifluoromethyl)pyrimidine

[0328] 2,4-Dichloro-5-(trifluoromethyl)pyrimidine Cpd-27A (3.24 g, 14.8 mmol), pinacol m-methylphenylboronic acid (3.24 g, 14.8 mmol), palladium dichloride bis(triphenylphosphine) (1.04 g, 1.48 mmol), and sodium carbonate (2.36 g, 22.2 mmol) were dissolved in ethylene glycol (52 mL) and water (13 mL) and purged with nitrogen. The reaction was carried out at 40 °C for 12 hours. After the reaction was completed, the mixture was cooled to room temperature and filtered through diatomaceous earth. Water (50 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and rotary evaporated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give 2-chloro-4-m-tolyl-5-(trifluoromethyl)pyrimidine Cpd-27B (600 mg, white oil). Yield: 15%.

[0329] MS m / z(ESI): 273[M+1] + .

[0330] 1 H NMR (400MHz, CDCl3) δ8.93 (s, 1H), 7.39 (dd, J = 16.0, 9.5Hz, 4H), 2.43 (s, 3H).

[0331] Referring to steps four through six of Example 13, compound Cpd-07D was reacted with compound Cpd-27B to obtain compound (S)-2-amino-3-(3-methoxy-4-((4-m-tolyl-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid Cpd-27.

[0332] MS m / z(ESI): 447[M+1] + .

[0333] 1 H NMR (400MHz, DMSO-d6) δ9.15(s,1H),8.77(s,1H),8.42(d,J=3.5Hz,3H),7.72(d,J=8.1Hz,1H),7.42-7.27(m,4H),7.06( d,J=1.5Hz,1H),6.83(dd,J=8.1,1.3Hz,1H),4.26-4.10(m,1H),3.83(s,3H),3.13(qd,J=14.3,6.2Hz,2H),2.37(s,3H).

[0334] Example 18

[0335] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethoxyphenyl)propionic acid

[0336]

[0337] first step

[0338] Preparation of 4-bromo-2-ethoxy-1-nitrobenzene

[0339] 5-Bromo-2-nitrophenol Cpd-28A (1.0 g, 4.6 mmol), iodoethane (1.0 g, 6.9 mmol), and cesium carbonate (3.0 g, 9.2 mmol) were added to acetone (10 mL) and reacted at 65 °C for 3 hours. After the reaction was complete, water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to give 4-bromo-2-ethoxy-1-nitrobenzene Cpd-28B (1.1 g, yellow solid), yield: 91%.

[0340] 1 H NMR (400MHz, DMSO-d6) δ7.85(d,J=8.6Hz,1H),7.61(d,J=1.9Hz,1H),7.34(dd,J=8.6,1.9Hz,1H),4.27(q,J=7.0Hz,2H),1.35(t,J=7.0Hz,3H).

[0341] Referring to steps two through six of Example 13, the difference is that compound Cpd-16B is replaced with compound Cpd-28B to obtain compound (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethoxyphenyl)propionic acid Cpd-28.

[0342] MS m / z(ESI): 426[M+1] + .

[0343] 1 H NMR(400MHz,DMSO-d6)δ13.84(s,1H),8.78(s,1H),8.38(s,5H),7.89(s,1H),7.05(s,1H),6.86(d ,J=8.2Hz,1H),4.27–4.00(m,2H),3.11(s,2H),2.88(s,1H),1.49–1.23(m,4H),0.94–0.58(m,4H).

[0344] Example 19

[0345] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethynylphenyl)propionic acid

[0346]

[0347] first step

[0348] Preparation of methyl (S)-3-(4-amino-3-iodophenyl)-2-((tert-butoxycarbonyl)amino)propionate

[0349] Methyl (S)-3-(4-aminophenyl)-2-((tert-butoxycarbonyl)amino)propionate Cpd-32A (1.8 g, 6.1 mmol) was dissolved in N,N-dimethylformamide (30 mL), and N-iodosuccinimide (1.5 g, 6.7 mmol) was added. The mixture was stirred at 70 °C for 16 hours. Ethyl acetate (100 mL) was added to the reaction solution, and the mixture was washed once with water (100 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give methyl (S)-3-(4-amino-3-iodophenyl)-2-((tert-butoxycarbonyl)amino)propionate Cpd-32B (1.8 g, yellow oil), yield: 67%.

[0350] MS m / z(ESI): 443[M+23] + .

[0351] Step 2

[0352] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-iodophenyl)propionate

[0353] Methyl (S)-3-(4-amino-3-iodophenyl)-2-((tert-butoxycarbonyl)amino)propionate Cpd-32B (1.3 g, 3.1 mmol) was dissolved in n-butanol (20 mL), and 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidine-4-amine (0.74 g, 3.1 mmol) and trifluoroacetic acid (0.18 g, 1.6 mmol) were added. The mixture was stirred at 80 °C for 16 hours. Ethyl acetate (150 mL) was added to the reaction solution. The organic phase was washed successively with water and saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-iodophenyl)propionate Cpd-32C (0.75 g, pale yellow oil), yield: 35%.

[0354] MS m / z(ESI): 622[M+1] + .

[0355] Step 3

[0356] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-((trimethylsilyl)ethynyl)phenyl)propionate

[0357] Methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-iodophenyl)propionate Cpd-32C (200 mg, 0.32 mmol) was dissolved in acetonitrile (5 mL), and ethynyltrimethylsilane (35 mg, 0.35 mmol), triethylenediamine (180 mg, 1.6 mmol), allyl palladium(II) dimer (47 mg, 0.13 mmol), and tri-tert-butylphosphine (13 mg, 0.06 mmol) were added. The mixture was stirred at 40 °C for 4 hours under nitrogen protection. The reaction solution was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-((trimethylsilyl)ethynyl)phenyl)propionate Cpd-32D (60 mg, white solid), yield: 47%.

[0358] MS m / z(ESI): 592 [M+1] + .

[0359] Step 4

[0360] Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethynylphenyl)propionic acid

[0361] Methyl propionate Cpd-32D (80 mg, 0.13 mmol) was dissolved in a 1 / 4 (2 mL) mixture of tetrahydrofuran and water. Lithium hydroxide monohydrate (11 mg, 0.27 mmol) was added to the solution, and the mixture was stirred at room temperature for 1 hour. The reaction solution was extracted with ethyl acetate (20 mL × 2). The aqueous phase was adjusted to pH 3 to 4 with 20% hydrochloric acid, extracted with ethyl acetate (50 mL × 2), washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethynylphenyl)propionic acid Cpd-32E (60 mg, yellow oil), yield: 87%.

[0362] MS m / z(ESI): 506 [M+1] + .

[0363] Step 5

[0364] Preparation of (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethynylphenyl)propionic acid

[0365] (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethynylphenyl)propionic acid Cpd-32E (50 mg, 0.10 mmol) was added to a hydrochloric acid / dioxane (4 N, 2 mL) solution and stirred at room temperature for 1 hour. The reaction solution was concentrated, and the residue was lyophilized with water to obtain (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-ethynylphenyl)propionic acid Cpd-32 (40 mg), 99%

[0366] MS m / z(ESI): 406[M+1] + .

[0367] 1H NMR (400MHz, DMSO-d6) δ8.89(s,1H),8.37(d,J=27.3Hz,5H),7.85(s,1H),7.44(s,1H),7.35(d,J=8.6 Hz,1H),4.70(s,1H),4.20(d,J=5.5Hz,1H),3.15-3.06(m,2H),2.85-2.83(m,1H),0.81-0.69(m,4H).

[0368] Example 20

[0369] (S)-2-amino-3-(4-((5-chloro-4-((2-(dimethylphosphoryl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid

[0370]

[0371] Referring to steps four through six of Example 13, compound Cpd-07D was reacted with compound 2-chloro-N-(2-(dimethylphosphoryl)phenyl)-5-chloropyrimidin-4-amine to finally obtain compound (S)-2-amino-3-(4-((5-chloro-4-((2-(dimethylphosphoryl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-34.

[0372] Cpd-34:

[0373] MS m / z(ESI): 490[M+1] + .

[0374] 1 H NMR (400MHz, DMSO-d6) δ11.20(s,1H),8.50(dd,J=8.3,4.2Hz,1H),8.15(s,1H),8.06(s,1H),7.79(d,J=8.1Hz,1H),7.54(m,3H),7.16(t,J=7.2Hz,1 H),6.98(s,1H),6.79(d,J=8.1Hz,1H),3.82(s,3H),3.56(s,1H),3.16(dd ,J=14.2,4.2Hz,1H),2.85(dd,J=14.3,8.3Hz,1H),1.78(d,J=13.5Hz,6H).

[0375] Example 21

[0376] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-(deuterated methoxy)phenyl)propionic acid:

[0377]

[0378] first step

[0379] Preparation of 4-bromo-2-(deuterated methoxy)-1-nitrobenzene

[0380] 5-Bromo-2-nitrophenol Cpd-35A (1.0 g, 4.6 mmol), deuterated iodomethane (1.0 g, 6.9 mmol), and cesium carbonate (3.0 g, 9.2 mmol) were added to acetone (10 mL) and reacted at 65 °C for 3 hours. After the reaction was complete, water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to give 4-bromo-2-(deuterated methoxy)-1-nitrobenzene Cpd-35B (1.0 g, yellow solid), yield: 87%.

[0381] 1 H NMR (400MHz, DMSO-d6) δ7.85 (d, J = 8.6 Hz, 1H), 7.61 (d, J = 1.9 Hz, 1H), 7.34 (dd, J = 8.6, 1.9 Hz, 1H).

[0382] Referring to the synthesis steps of Example 13, except that compound Cpd-16B was replaced with compound Cpd-35B to obtain compound (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-(deuterated methoxy)phenyl)propionic acid Cpd-35.

[0383] MS m / z(ESI): 415[M+1] + .

[0384] 1 H NMR (400MHz, DMSO-d6) δ13.85(s,1H),8.80(s,1H),8.40(d,J=36.3Hz,5H),7.87(s,1H),7.05(d,J=1.3Hz,1H),6. 86(dd,J=8.3,1.5Hz,1H),4.19(s,1H),3.11(qd,J=14.3,6.4Hz,2H),2.87(s,1H),1.25(s,1H),0.86–0.68(m,4H).

[0385] Example 22

[0386] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)tert-butyl propionate

[0387]

[0388] first step

[0389] Preparation of tert-butyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate

[0390] (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-07F (120 mg, 0.23 mmol) was dissolved in dichloromethane (5 mL). Tert-butanol (173 mg, 2.30 mmol) and 4-dimethylaminopyridine (3 mg, 0.02 mmol) were added at 0 °C and reacted for 10 minutes. N,N'-dicyclohexylcarbodiimide (97 mg, 0.46 mmol) was then added at 0 °C. The reaction mixture was heated from 0 °C to room temperature and reacted overnight. After the reaction was complete, the reaction solution was filtered with diatomaceous earth, and the crude product was obtained by rotary evaporation. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate tert-butyl ester Cpd-36B (140 mg, white solid), yield: 95%.

[0391] MS m / z(ESI): 568 [M+1] + .

[0392] Step 2

[0393] Preparation of tert-butyl (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate

[0394] A solution of dioxane (5 mL, 4 mol / L) hydrogen chloride was added to (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate tert-butyl ester Cpd-36B (100 g, 0.24 mmol). The reaction solution was reacted at 25 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated to give the hydrochloride of (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate tert-butyl ester Cpd-36 (15 mg), yield: 14%.

[0395] MS m / z(ESI): 468[M+1] + .

[0396] 1 H NMR (400MHz, CD3OD) δ8.31(s,2H),7.12(s,1H),6.98(d,J=8.1Hz,1H),4.27(t,J=6.9Hz,1H),3.99 (s,3H),3.23(d,J=5.1Hz,2H),2.99(s,1H),1.46(s,9H),0.96(d,J=5.6Hz,2H),0.86-0.75(m,2H).

[0397] Example 23

[0398] (S)-((2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionyl)oxy)neopentate methyl ester

[0399]

[0400] first step

[0401] Preparation of methyl neopentanoate (S)-((2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionyl)oxy)neopentate

[0402] (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionic acid Cpd-07F (250 mg, 0.48 mmol), methyl iodide pentovalinate (142 mg, 0.58 mmol), and potassium carbonate (134 mg, 0.97 mmol) were added to N,N-dimethylformamide (5 mL). The reaction was carried out at room temperature for 8 hours. After the reaction was completed, the reaction solution was added to water (10 mL), extracted with ethyl acetate (20 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. Methyl (S)-((2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionyl)oxy)neopentaate Cpd-37B (320 mg, white oil) was obtained in 99% yield.

[0403] MS m / z(ESI): 626[M+1] + .

[0404] Step 2

[0405] Preparation of (S)-((2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionyl)oxy)neopentaic acid methyl ester

[0406] Methyl (S)-((2-((tert-Butoxycarbonyl)amino)-3-(4-(((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionyl)oxy)neopentaate Cpd-37B (320 mg, 0.51 mmol)) was dissolved in hydrochloric acid / dioxane (4 M, 10 mL) and stirred at room temperature for 1 hour. The reaction solution was concentrated and dried under vacuum to give methyl (S)-((2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionyl)oxy)neopentaate Cpd-37 (250 mg) hydrochloride, yield: 88%.

[0407] MS m / z(ESI): 526 [M+1] + .

[0408] 1H NMR (400MHz, DMSO-d6) δ8.61(s,3H),8.36(s,2H),7.08(s,1H),6.84(d,J=8.2Hz,1H),5.84(d,J=5.9Hz,1H) ,5.78(d,J=5.9Hz,1H),4.46(s,2H),3.90(s,3H),3.11(s,2H),2.88(s,1H),1.15(s,9H),0.89–0.65(m,4H).

[0409] Example 24

[0410] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)heptyl propionate

[0411]

[0412] Referring to the synthesis steps of Example 22, except that heptanol was used instead of tert-butanol, compound (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propionate hepta-ester Cpd-38 was obtained.

[0413] MS m / z(ESI): 510 [M+1] + .

[0414] 1 H NMR (400MHz, CD3OD) δ8.29(s,2H),7.07(s,1H),6.94(d,J=8.2Hz,1H),4.37(t,J=7.1Hz,1H),4.19(t,J=6.6Hz,2H),3.98(s,3H),3.24(dd,J =6.2,4.4Hz,2H),3.05-2.89(m,1H),1.66-1.54(m,2H),1.27(s,8H),0.95(dd,J=7.1,5.4Hz,2H),0.87(t,J=6.9Hz,3H),0.83-0.73(m,2H).

[0415] Example 25

[0416] (S)-2-amino-3-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propionic acid

[0417]

[0418] first step

[0419] Preparation of (S)-2-((benzyloxycarbonyl)amino)-3-(3-methyl-4-nitro-1H-pyrazol-1-yl)benzyl propionate

[0420] 3-Methyl-4-nitro-1H-pyrazole Cpd-39A (0.8 g, 6.5 mmol), (R)-2-((benzyloxycarbonyl)amino)-3-iodopropionate benzyl ester (3.0 g, 6.5 mmol), and cesium carbonate (2.7 g, 7.8 mmol) were dissolved in N,N-dimethylformamide (20 mL) and stirred overnight at room temperature. After the reaction was complete, water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and rotary evaporated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give (S)-2-((benzyloxycarbonyl)amino)-3-(3-methyl-4-nitro-1H-pyrazole-1-yl)propionate Cpd-39B (2.2 g, white oil), yield: 70%.

[0421] MS m / z(ESI): 439[M+1] + .

[0422] 1 H NMR(400MHz, DMSO-d6)δ8.72(s,1H),7.96(dd,J=11.9,8.5Hz,1H),7.37-7.30(m,10H),5.15(s,2H ), 5.02 (d, J = 3.7Hz, 2H), 4.68 (dt, J = 8.3, 4.2Hz, 1H), 4.03 (q, J = 7.1Hz, 2H), 1.17 (t, J = 7.1Hz, 3H).

[0423] Step 2

[0424] Preparation of (S)-3-(4-amino-3-methyl-1H-pyrazol-1-yl)-2-((benzyloxycarbonyl)amino)propionate

[0425] (S)-2-((benzyloxycarbonyl)amino)-3-(3-methyl-4-nitro-1H-pyrazol-1-yl)propionate Cpd-39B (1.0 g, 2.3 mmol), iron powder (0.96 g, 17 mmol), and saturated aqueous solution of ammonium chloride (0.18 g, 3.5 mmol) were dissolved in ethanol (10 mL) and refluxed for 2 hours. After the reaction was completed, the mixture was cooled to room temperature and filtered through diatomaceous earth. Water (5 mL) and ethyl acetate (10 mL × 3) were added for extraction. The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 2) to give (S)-3-(4-amino-3-methyl-1H-pyrazol-1-yl)-2-((benzyloxycarbonyl)amino)propionate Cpd-39C (0.4 g, brown oil), yield: 42%.

[0426] MS m / z(ESI): 409[M+1] + .

[0427] Step 3

[0428] Preparation of (S)-2-((benzyloxycarbonyl)amino)-3-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)benzyl propionate

[0429] (S)-3-(4-amino-3-methyl-1H-pyrazol-1-yl)-2-((benzyloxycarbonyl)amino)propionate Cpd-39C (0.4 g, 0.98 mmol), 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (0.2 g, 0.98 mmol) were dissolved in n-butanol (10 mL), and trifluoroacetic acid (0.12 g, 0.98 mmol) was added. The mixture was reacted at 90 °C for 1 hour. After the reaction was completed, the mixture was cooled to room temperature, quenched with water (5 mL), extracted with ethyl acetate (10 mL × 3), washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give (S)-2-((benzyloxycarbonyl)amino)-3-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propionate Cpd-39D (0.29 g, 0.5 mmol), yield: 51%.

[0430] MS m / z(ESI): 584 [M+1] + .

[0431] Step 4

[0432] Preparation of (S)-2-amino-3-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propionic acid

[0433] (S)-2-((benzyloxycarbonyl)amino)-3-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propionate Cpd-39D (0.2 g, 0.34 mmol) and palladium on carbon (0.04 g) were added to methanol (10 mL) and purged with hydrogen. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, and the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by reverse preparation (acetonitrile / water (trifluoroacetic acid 1‰)) to obtain (S)-2-amino-3-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propionate Cpd-39 (0.08 g, 0.22 mmol), yield: 67%.

[0434] MS m / z(ESI): 360[M+1] + .

[0435] 1 H NMR (400MHz, MeOD) δ7.98-7.72(m,2H),4.61(s,2H),4.36(s,1H),3.05(s,3H),2.20(s,3H).

[0436] Example 26

[0437] (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionic acid

[0438]

[0439] first step

[0440] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(3-methyl-4-nitro-1H-pyrazol-1-yl)propionate

[0441] 3-Methyl-4-nitro-1H-pyrazole Cpd-40A (1.9 g, 15.2 mmol), methyl (R)-2-((tert-butyloxycarbonyl)amino)-3-iodopropionate (5.0 g, 15.2 mmol), and cesium carbonate (6.0 g, 18.2 mmol) were dissolved in N,N-dimethylformamide (40 mL) and stirred overnight at room temperature. After the reaction was complete, water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and rotary evaporated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give methyl (S)-2-((tert-butyloxycarbonyl)amino)-3-(3-methyl-4-nitro-1H-pyrazole-1-yl)propionate Cpd-40B (3.5 g, white oil), yield: 70%.

[0442] MS m / z(ESI): 329[M+1] + .

[0443] Step 2

[0444] Preparation of methyl (S)-3-(4-amino-3-methyl-1H-pyrazol-1-yl)-2-((tert-butoxycarbonyl)amino)propionate

[0445] Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(3-methyl-4-nitro-1H-pyrazol-1-yl)propionate Cpd-40B (0.5 g, 1.5 mmol) and palladium on carbon (0.05 g) were added to methanol (10 mL), and the mixture was purged with hydrogen. The reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was filtered through diatomaceous earth, and the filtrate was evaporated to dryness to obtain crude (S)-3-(4-amino-3-methyl-1H-pyrazol-1-yl)-2-((tert-Butoxycarbonyl)amino)propionate Cpd-40C (0.4 g, pink oil), yield: 88%.

[0446] MS m / z(ESI): 299[M+1] + .

[0447] Step 3

[0448] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionate

[0449] Methyl (S)-3-(4-amino-3-methyl-1H-pyrazol-1-yl)-2-((tert-butoxycarbonyl)amino)propionate Cpd-40C (0.40 g, 1.3 mmol), 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidin-4-amine (0.32 g, 1.3 mmol) were dissolved in n-butanol (8 mL), and trifluoroacetic acid (0.15 g, 1.3 mmol) was added. The mixture was reacted at 90 °C for 1 hour. After the reaction was completed, the mixture was cooled to room temperature, quenched with water (5 mL), extracted with ethyl acetate (10 mL × 3), washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionate Cpd-40D (0.2 g, 0.4 mmol), yield: 30%.

[0450] MS m / z(ESI): 500[M+1] + .

[0451] Step 4

[0452] Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionic acid

[0453] Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionate Cpd-40D (0.2 g, 0.4 mmol) was dissolved in tetrahydrofuran (5 mL), and saturated lithium hydroxide aqueous solution (0.5 mL) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was completed, the pH was adjusted to 4 with dilute hydrochloric acid, and the mixture was extracted with a mixed solvent (dichloromethanol / methanol = 10 / 1). The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography to obtain (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionic acid Cpd-40E (0.15 g, white solid), yield: 77%.

[0454] MS m / z(ESI): 486[M+1] + .

[0455] Step 5

[0456] Preparation of (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionic acid

[0457] (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionic acid Cpd-40E (0.15 g, 0.31 mmol) was dissolved in HCl / 1,4-dioxane (15 mL) and stirred at 25 °C for 1 hour. After the reaction was completed, the filtrate was evaporated to dryness to obtain the crude product, which was purified by reverse preparation (acetonitrile / water (formic acid 1‰)) to obtain (S)-2-amino-3-(4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)propionic acid Cpd-40 (46 mg), yield: 39%.

[0458] MS m / z(ESI): 386[M+1] + .

[0459] 1 H NMR(400MHz,DMSO-d6)δ9.14(s,1H),8.22(s,1H),8.12(s,1H),7.61(s,3H),7.02(s,1H),4.39(s,1H),4 .20(dd,J=14.4,8.0Hz,1H),3.55(d,J=4.8Hz,1H),2.92(s,1H),2.19(s,3H),0.81(s,2H),0.64(s,2H).

[0460] Example 27

[0461] (S)-2-amino-3-(3-cyclopropyl-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propionic acid

[0462]

[0463] Referring to the synthesis steps of Example 26, except that 3-methyl-4-nitro-1H-pyrazole Cpd-40A was replaced with compound 3-cyclopropyl-4-nitro-1H-pyrazole Cpd-41A as the starting compound, to obtain compound (S)-2-amino-3-(3-cyclopropyl-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazole-1-yl)propionic acid Cpd-41.

[0464] MS m / z(ESI): 412[M+1]+ .

[0465] 1 H NMR (400MHz, DMSO-d6) δ9.18 (s, 1H), 8.29–8.09 (m, 3H), 7.02 (s, 1H), 4.36 (d, J = 13.2Hz, 1H), 4. 15(dd,J=14.4,7.8Hz,1H),3.49(d,J=4.8Hz,2H),2.93(s,1H),2.18(s,1H),0.80–0.61(m,8H).

[0466] Example 28

[0467] (S)-2-amino-3-(4-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionic acid

[0468]

[0469] first step

[0470] Preparation of (3S,4S)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester

[0471] (3S,4R)-3-fluoro-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester Cpd-42A (2.0 g, 91 mmol), 4-nitro-1H-pyrazole (1.1 g, 10 mmol), and triphenylphosphine (3.6 g, 14 mmol) were dissolved in tetrahydrofuran (100 mL) under nitrogen protection. Diisopropyl azodicarbonate (2.8 g, 14 mmol) was added dropwise at 0 °C. After the addition was complete, the mixture was stirred overnight at 35 °C. After the reaction was complete, the mixture was concentrated under reduced pressure. Dichloromethane (100 mL) was added to the residue, and the mixture was washed once with water. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give compound (3S,4S)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester Cpd-42B (2.6 g, pale yellow oil), yield: 86%.

[0472] 1 H NMR (400MHz, CDCl3) δ8.27(s,1H),8.13(s,1H),4.83-4.69(m,2H),4.28(dt,J=10.5,6.3Hz,2H),2.87(s,2H),2.28-2.14(m,2H),1.49(s,9H).

[0473] Step 2

[0474] Preparation of tert-butyl (3S,4S)-4-(5-chloro-4-nitro-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid

[0475] (3S,4S)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester Cpd-42B (2.6 g, 8.2 mmol) was dissolved in tetrahydrofuran (50 mL) under nitrogen protection, and a tetrahydrofuran solution of bis(trimethylsilylamino)lithium (16.4 mL, 1 mol / L, 16.4 mmol) was added dropwise at -78 °C. After the addition was complete, the mixture was stirred at -78 °C for 40 minutes. Subsequently, a tetrahydrofuran solution of hexachloroethane (3.9 g, 16.4 mmol) was added dropwise at -78 °C. After the addition was complete, the mixture was stirred at -78 °C for 30 minutes. After the reaction was complete, saturated ammonium chloride (50 mL) was added to quench the reaction mixture. The mixture was extracted with ethyl acetate (100 mL × 2), and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain (3S,4S)-4-(5-chloro-4-nitro-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester Cpd-42C (2.5 g, pale yellow oil), yield: 83%.

[0476] 1 H NMR (400MHz, CDCl3) δ8.24(s,1H),4.97-4.78(m,1H),4.57-4.51(m,2H),4.27(s ,1H),2.90(d,J=10.8Hz,2H),2.26-2.18(m,1H),2.00-1.98(m,1H),1.49(s,9H).

[0477] Step 3

[0478] Preparation of (3S,4S)-4-(4-amino-5-chloro-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester

[0479] (3S,4S)-4-(5-chloro-4-nitro-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester Cpd-42C (2.4 g, 6.9 mmol) was dissolved in a mixed solvent (60 mL, ethanol / water = 5 / 1), and reduced iron powder (2.9 g, 51 mmol) and ammonium chloride (1.1 g, 21 mmol) were added. The mixture was stirred at 75 °C for 2 hours. After the reaction was completed, the reaction solution was filtered through diatomaceous earth while hot, concentrated under reduced pressure, and extracted with ethyl acetate (200 mL) to the residue. The residue was washed once with water, and the organic layer was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain (3S,4S)-4-(4-amino-5-chloro-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester Cpd-42D (1.7 g, yellow oil), yield: 74%.

[0480] MS m / z(ESI): 319[M+1] + .

[0481] Step 4

[0482] Preparation of tert-butyl (3S,4S)-4-(5-chloro-4-((4-methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid

[0483] (3S,4S)-4-(4-amino-5-chloro-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester Cpd-42D (1.6 g, 5.0 mmol) was dissolved in ethylene glycol monomethyl ether (30 mL), and 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (2.1 g, 10 mmol) and trifluoroacetic acid (0.57 g, 5.0 mmol) were added. The mixture was stirred at room temperature for 16 hours. Ethyl acetate (500 mL) was added to the reaction solution. The organic phase was washed successively with water and saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain (3S,4S)-4-(5-chloro-4-((4-methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester Cpd-42E (1.9 g, pale yellow oil), yield: 72%.

[0484] MS m / z(ESI): 494[M+1] + .

[0485] Step 5

[0486] N 2-(5-chloro-1-((3S,4S)-3-fluoropiperidin-4-yl)-1H-pyrazole-4-yl)-N 4 Preparation of 2,4-methyl-5-)trifluoromethyl)pyrimidine-2,4-diamine

[0487] In a mixture of (3S,4S)-4-(5-chloro-4-((4-methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-carboxylic acid tert-butyl ester Cpd-42E (1.9 g, 3.8 mmol) and dichloromethane (10 mL), trifluoroacetic acid (5 mL) was added, and the mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated under reduced pressure, and the pH of the residue was adjusted to 8-9 with saturated sodium carbonate solution. The residue was extracted with ethyl acetate (200 mL × 2), and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain N. 2 -(5-chloro-1-((3S,4S)-3-fluoropiperidin-4-yl)-1H-pyrazole-4-yl)-N 4 1,4-(methyl-5-)trifluoromethyl)pyrimidine-2,4-diamine Cpd-42F (1.6 g, pale yellow solid), yield: 95%.

[0488] MS m / z(ESI): 394[M+1] + .

[0489] Step 6

[0490] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionate

[0491] N 2 -(5-chloro-1-((3S,4S)-3-fluoropiperidin-4-yl)-1H-pyrazole-4-yl)-N 45-(methyl-5-)trifluoromethyl)pyrimidine-2,4-diamine Cpd-42F (500 mg, 1.3 mmol) was dissolved in dioxane (10 mL), and N-(tert-butoxycarbonyl)-L-4-bromophenylalanine methyl ester (1.4 g, 3.8 mmol), cesium carbonate (495 mg, 1.5 mmol) and XPhos Pd G3 (214 mg, 0.25 mmol) were added. The mixture was reacted at 105 °C for 16 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth. Ethyl acetate (50 mL) was added to the filtrate, and the mixture was washed once with water. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionate Cpd-42G (100 mg, pale yellow oil), yield: 11%.

[0492] MS m / z(ESI): 671[M+1] + .

[0493] Step 7

[0494] Preparation of methyl (S)-2-amino-3-(4-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionate

[0495] Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(4-(((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionate Cpd-42G (90 mg, 0.13 mmol) was dissolved in dioxane (0.1 mL), and HCl / dioxane (2 mL, 4 M) was added. The mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to give methyl (S)-2-amino-3-(4-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionate Cpd-42H (70 mg, pale yellow solid), yield: 87%.

[0496] MS m / z(ESI): 571 [M+1] + .

[0497] Step 8

[0498] Preparation of (S)-2-amino-3-(4-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionic acid

[0499] Preparation of methyl (S)-2-amino-3-(4-(((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionate: Cpd-42H (70 mg, 0.12 mmol) was dissolved in a mixed solvent (1 mL, methanol / water = 10 / 1), and lithium hydroxide (6 mg, 0.25 mmol) was added. The mixture was stirred at room temperature for 1 hour. The reaction solution was adjusted to pH 3 to 4 with 20% hydrochloric acid. The resulting solution was then directly purified by reverse-phase preparation (ACN / H2O (0.1% TFA)) to obtain (S)-2-amino-3-(4-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionic acid Cpd-42 (10 mg), yield: 14%.

[0500] MS m / z(ESI): 557[M+1] + .

[0501] H NMR (400MHz, DMSO-d6) δ9.00(s,1H),8.09(s,1H),7.90(s,1H),7.14(d,J=8.2Hz,2H),7.03(s,1H),6.97(d,J=8.3Hz,2H),5.05-4.86(m,1H) ,4.68-4.59(m,1H),4.13(d,J=11.8Hz,1H),3.77(s,1H),3.32(s,1H), 3.03-2.97(m,4H),2.82(s,3H),2.21-2.13(m,1H),2.11-2.00(m,1H).

[0502] Example 29

[0503] (S)-2-amino-3-(3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionic acid

[0504]

[0505] Referring to the synthetic steps of Example 28, compound Cpd-42F was first prepared. In step six, N-(tert-butoxycarbonyl)-L-3-bromophenylalanine methyl ester was replaced with (S)-3-(4-bromophenyl)-2-(tert-butoxycarbonyl)amino)propionate methyl ester to obtain compound (S)-2-amino-3-(3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)phenyl)propionate Cpd-43 (35 mg), yield: 49%.

[0506] MS m / z(ESI): 557[M+1] + .

[0507] 1 H NMR (400MHz, DMSO-d6) δ10.50(s,1H),8.45(d,J=38.1Hz,4H),8.09(d,J=41.9Hz,2H),7.20(t,J=7.8Hz,1H),7.02-6.97(m,2H),6.73(d,J= 7.4Hz,1H),4.96(d,J=52.1Hz,1H),4.76-4.71(m,,1H),4.19(s,2H),3.85-3.82(m,1H),3.15-3.03(m,4H),2.92(s,3H),2.22-2.10(m,2H).

[0508] Example 30

[0509] (S)-2-amino-3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid

[0510]

[0511] first step

[0512] Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid

[0513] N 2 -(5-chloro-1-((3S,4S)-3-fluoropiperidin-4-yl)-1H-pyrazole-4-yl)-N 4(-Methyl-5-)trifluoromethyl)pyrimidin-2,4-diamine Cpd-42F (200 mg, 0.51 mmol) was dissolved in dichloromethane (10 mL), and compound F-1 (142 mg, 0.76 mmol) was added. The reaction was carried out at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give (S)-2-((tert-butoxycarbonyl)amino)-3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid Cpd-44B (60 mg, pale yellow oil), yield: 19%.

[0514] MS m / z(ESI): 581 [M+1] + .

[0515] 1 H NMR (400MHz, DMSO-d6) δ12.63(s,1H),8.99(s,1H),8.09(s,1H),7.87(s,1H),7.00(d,J=11 .3Hz,2H),4.88-4.70(m,1H),4.40-4.35(m,1H),4.16-4.04(m,4H),2.88-2.75(m,6H),2.34 -2.29(m,2H),2.02-1.89(m,1H),1.40(s,9H).

[0516] Step 2

[0517] (S)-2-amino-3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid

[0518] (S)-2-((tert-Butoxycarbonyl)amino)-3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid Cpd-44B (50 mg, 0.09 mmol) was dissolved in dioxane (0.5 mL), and HCl / dioxane (2 mL, 4 M) was added. The mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated and dried under vacuum to give (S)-2-amino-3-((3S,4S)-4-(5-chloro-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid Cpd-44 (27 mg) hydrochloride, yield: 50%.

[0519] MS m / z(ESI): 481[M+1] + .

[0520] 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),8.44(s,4H),8.02(s,1H),5.09(d,J=48.5Hz,1H),4.51-4.30(m ,2H),3.77-3.49(m,1H),3.05-2.91(m,6H),2.67(s,1H),2.19(dd,J=17.8,10.2Hz,1H),2.01(s,1H).

[0521] Example 31

[0522] (S)-2-amino-3-((3S,4S)-4-(5-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid

[0523]

[0524] first step

[0525] Preparation of tert-butyl (3S,4S)-4-(5-chloro-4-((4-cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid

[0526] (3S,4S)-4-(4-amino-5-chloro-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester Cpd-42D (500 mg, 1.6 mmol) was dissolved in ethylene glycol monomethyl ether (10 mL), and 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidine-4-amine (447 mg, 1.9 mmol) and trifluoroacetic acid (178 mg, 1.6 mmol) were added. The mixture was stirred at room temperature for 16 hours. Ethyl acetate (150 mL) was added to the reaction solution. The organic phase was washed successively with water and saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain (3S,4S)-4-(5-chloro-4-((4-cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester Cpd-45B (580 mg, pale yellow oil), yield: 70%.

[0527] MS m / z(ESI): 520 [M+1] + .

[0528] Step 2: Referring to Step 5 of Example 28, deprotect compound Cpd-45B from Boc to obtain compound N. 2 -(5-chloro-1-((3S,4S)-3-fluoropiperidin-4-yl)-1H-pyrazole-4-yl)-N 4 -Cyclopropyl-5-(trifluoromethyl)pyrimidine-2,4-diamine Cpd-45C (180 mg, pale yellow solid), yield: 98%.

[0529] MS m / z(ESI): 420[M+1] + .

[0530] Steps three and four are performed in accordance with steps one and two of Example 30, to obtain the hydrochloride salt of compound (S)-2-amino-3-((3S,4S)-4-(5-chloro-4-((4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)-3-fluoropiperidin-1-yl)propionic acid Cpd-45 (30 mg), yield: 50%.

[0531] MS m / z(ESI): 507 [M+1] + .

[0532] 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.49(d,J=22.2Hz,4H),8.17(s,1H),5.15(d,J=48.9Hz,1H),4.61-4.56(m,1H ),4.42(s,1H),3.83-3.73(m,1H),3.30-3.11(m,3H),2.90(s,3H),2.30(m,1H),2.09(s,1H),0.74(d,J=3.9Hz,4H).

[0533] Example 32

[0534] (S)-2-amino-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0535]

[0536] first step

[0537] Preparation of 1-bromo-5-chloro-2-methoxy-4-nitrobenzene

[0538] 2-Bromo-4-chloro-5-nitrophenol Cpd-54A (1.0 g, 3.9 mmol), methyl iodoform (1.0 g, 5.9 mmol), and cesium carbonate (3.0 g, 7.9 mmol) were added to acetone (20 mL) and reacted at 65 °C for 5 hours. After the reaction was complete, water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give 1-bromo-5-chloro-2-methoxy-4-nitrobenzene Cpd-54B (1.0 g, yellow solid), yield: 97%.

[0539] 1 H NMR (400MHz, DMSO-d6) δ8.11(s,1H),7.82(s,1H),3.95(s,3H).

[0540] Step 2

[0541] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-nitrobenzene)propionate

[0542] 1-Bromo-5-chloro-2-methoxy-4-nitrobenzene Cpd-54B (1.0 g, 3.8 mmol), bis(triphenylphosphine)palladium dichloride (0.27 g, 0.38 mmol), cuprous iodide (0.17 g, 0.92 mmol), and N,N-dimethylformamide (20 mL) were added to a three-necked flask. After purging with nitrogen three times, the flask was preheated in an oil bath at 80 °C for 10 minutes. Subsequently, a solution of (R)-2-(tert-butoxycarbonyl)amino)-3-iodopropionate methyl iodide zinc iodide in N,N-dimethylformamide was slowly added dropwise using a syringe. The reaction mixture was stirred at 80 °C for 0.5 hours. After the reaction was completed, the reaction mixture was filtered through a diatomaceous earth liner, then poured into water (30 mL), extracted with ethyl acetate (50 mL × 3), combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-nitrobenzene)propionate Cpd-54C (700 mg, yellow solid), yield: 47%.

[0543] MS m / z(ESI): 411[M+23] + .

[0544] Step 3

[0545] Preparation of methyl (S)-3-(4-amino-5-chloro-2-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propionate

[0546] Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-nitrobenzene)propionate Cpd-54C (700 mg, 1.8 mmol), ammonium chloride (144 mg, 2.7 mmol), and iron powder (753 mg, 13 mmol) were added to a mixed solvent (ethanol / water = 5 / 1, 18 mL) and reacted at 75 °C under a nitrogen atmosphere for 1 hour. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, the filtrate was concentrated and poured into water (30 mL), extracted with ethyl acetate (50 mL × 3), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain methyl (S)-3-(4-amino-5-chloro-2-methoxyphenyl)-2-((tert-Butoxycarbonyl)amino)propionate Cpd-54D (500 mg, orange-red solid), yield: 78%.

[0547] MS m / z(ESI): 381 [M+23] + .

[0548] Step 4

[0549] Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate

[0550] Methyl (S)-3-(4-amino-5-chloro-2-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propionate Cpd-54D (500 mg, 1.4 mmol), 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (357 mg, 1.7 mmol) were added to tert-butanol (10 mL), followed by the addition of trifluoroacetic acid (80 mg, 0.70 mmol) and dichloromethane (10 mL), and the mixture was purged with nitrogen three times. The reaction solution was reacted at 80 °C for 1 hour. After the reaction was complete, the reaction solution was concentrated, then ethyl acetate was added, stirred, filtered, and the filter cake was washed with ethyl acetate to obtain methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate Cpd-54E (360 mg, white solid), yield: 48%.

[0551] MS m / z(ESI): 534 [M+1] + .

[0552] Step 5

[0553] Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0554] Methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionate Cpd-54E (360 mg, 0.67 mmol) was dissolved in tetrahydrofuran (5 mL), and a solution of lithium hydroxide (49 mg, 2.0 mmol) in water (5 mL) was added dropwise. The mixture was reacted at room temperature for 1 hour. After the reaction was complete, the solution was poured into water (20 mL). The pH of the aqueous phase was adjusted to 4 with dilute hydrochloric acid (1 mol / L), and the solution was extracted with ethyl acetate (20 mL × 3). The organic phases were combined and concentrated to give (S)-2-((tert-butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid Cpd-54F (190 mg, white solid), yield: 54%.

[0555] MS m / z(ESI): 520 [M+1] + .

[0556] Step 6

[0557] Preparation of (S)-2-amino-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid

[0558] (S)-2-((tert-Butoxycarbonyl)amino)-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid Cpd-54F (190 mg, 0.37 mmol) was dissolved in a dioxane solution of hydrogen chloride (10 mL) and stirred at room temperature for 1 hour. The reaction solution was concentrated and dried under vacuum to give (S)-2-amino-3-(5-chloro-2-methoxy-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)propionic acid Cpd-54 (168 mg), yield: 95%.

[0559] MS m / z(ESI): 420[M+1] + .

[0560] 1H NMR(400MHz,DMSO-d6)δ13.73(s,1H),9.54(s,1H),8.35(s,4H),8.00(s,1H),7.67(s,1H), 7.35(s,1H),4.06(d,J=5.5Hz,1H),3.78(s,3H),3.19–2.96(m,2H),2.91(d,J=4.4Hz,3H).

[0561] Example 33

[0562] 2-Amino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionic acid

[0563]

[0564] first step

[0565] Preparation of 2,5-dimethoxy-4-nitrobobenzene

[0566] 2-Bromo-1,4-dimethoxybenzene Cpd-55A (1.0 g, 4.6 mmol) was dissolved in acetonitrile (2 mL), stirred at room temperature, and concentrated nitric acid (0.4 mL) was slowly added. The mixture was allowed to react at room temperature for 15 minutes. After the reaction was complete, water (10 mL) was added to the reaction solution, and the reaction solution was filtered to obtain a solid. The crude product was rotary evaporated to give 2,5-dimethoxy-4-nitrobromobenzene Cpd-55B (1.1 g, yellow solid), yield: 92%.

[0567] 1 H NMR (400MHz, DMSO-d6) δ7.68 (d, J = 1.9Hz, 1H), 7.64 (s, 1H), 3.91 (s, 3H), 3.87 (s, 3H).

[0568] Step 2

[0569] Preparation of methyl 2-(tert-Butoxycarbonyl)amino-3-(2,5-dimethoxy-4-nitrophenyl)propionate

[0570] Zinc powder (0.98 g, 15 mmol) and iodine (0.19 g, 0.75 mmol) were placed in a three-necked flask, purged with nitrogen, and heated to sublimate the iodine. 2,5-Dimethoxy-4-nitrobromobenzene Cpd-55B (0.4 g, 1.5 mmol) was dissolved in N,N-dimethylformamide (10 mL), and the solution was rapidly poured into the three-necked flask with stirring. Simultaneously, in another three-necked flask, cuprous iodide (0.07 g, 0.36 mmol), palladium dichloride bis(triphenylphosphine) dichloride (0.11 g, 0.15 mmol), and methyl(R)-2-((tert-butoxycarbonyl)amino)-3-iodopropionate (1.0 g, 3 mmol) were dissolved in N,N-dimethylformamide (10 mL) and purged with nitrogen. The mixture was reacted at 80 °C for 30 minutes. The zinc powder mixture was drawn up with a syringe and quickly injected into a three-necked flask at 80°C, and the reaction was continued for 1 hour. After the reaction was completed, the mixture was cooled to room temperature and filtered through diatomaceous earth. The solution was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give methyl 2-(tert-butoxycarbonyl)amino-3-(2,5-dimethoxy-4-nitrophenyl)propionate Cpd-55C (400 mg, yellow oil). Yield: 35%.

[0571] MS m / z(ESI): 385[M+1] + .

[0572] Step 3

[0573] Preparation of methyl 3-(4-amino-2,5-dimethoxyphenyl)-2-(tert-butoxycarbonyl)amino)propionate

[0574] Methyl 2-(tert-Butoxycarbonyl)amino-3-(2,5-dimethoxy-4-nitrophenyl)propionate Cpd-55C (400 mg, 1 mmol) was dissolved in methanol (10 mL), and palladium on carbon (109 mg, 0.1 mmol) was added. The reaction was carried out at room temperature for 1 hour under hydrogen protection. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, washed with methanol, and rotary evaporated to obtain crude methyl 3-(4-amino-2,5-dimethoxyphenyl)-2-(tert-Butoxycarbonyl)amino)propionate Cpd-55D (200 mg, yellow oil). Yield: 55%.

[0575] MS m / z(ESI): 355[M+1] + .

[0576] Step 4

[0577] Preparation of methyl 2-tert-butoxycarbonylamino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionate

[0578] Methyl 3-(4-amino-2,5-dimethoxyphenyl)-2-(tert-butoxycarbonyl)amino)propionate Cpd-55D (200 mg, 0.56 mol), 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidin-4-amine (159.61 mg, 0.67 mmol), and trifluoroacetic acid (32 mg, 0.28 mmol) were dissolved in n-butanol (10 mL) and reacted at 90 °C for 1 hour. After the reaction was completed, the mixture was cooled to room temperature and rotary evaporated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give methyl 2-tert-butoxycarbonylamino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionate Cpd-55E (100 mg, white solid). Yield: 34%.

[0579] MS m / z(ESI): 530 [M+1] + .

[0580] Step 5

[0581] Preparation of 2-tert-butoxycarbonylamino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionic acid

[0582] Methyl 2-tert-butoxycarbonylamino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionate Cpd-55E (100 mg, 0.18 mmol) was dissolved in tetrahydrofuran (5 mL), and lithium hydroxide aqueous solution (0.5 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the pH was adjusted to neutral with dilute hydrochloric acid, and the mixture was extracted with dichloromethanol / methanol (10 / 1). The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and rotary evaporated to obtain 2-tert-butoxycarbonylamino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionate Cpd-55F (70 mg, white solid), yield: 76%.

[0583] MS m / z(ESI): 516[M+1] + .

[0584] Step 6

[0585] Preparation of 2-amino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionic acid

[0586] 2-tert-Butoxycarbonylamino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionic acid Cpd-55F (70 mg, 0.13 mmol) was added to a dioxane solution of hydrogen chloride (4 mL, 4 mol / L). The reaction solution was reacted at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated and purified by reverse preparative reaction to obtain 2-amino-3-(2,5-dimethoxy-4-(4-methylamino)-5-trifluoromethylpyrimidin-2-ylamino)phenyl)propionic acid Cpd-55 (30 mg). Yield: 56%.

[0587] MS m / z(ESI): 416[M+1] + .

[0588] 1 H NMR(400MHz,CD3OD)δ8.29(s,1H),8.16(s,1H),8.10(s,1H),6.92(s,1H),3.90(s,3H),3.87(s, 3H),3.87–3.80(m,2H),3.37(dd,J=14.3,4.1Hz,1H),3.09(s,3H),2.95(dd,J=14.3,8.8Hz,1H).

[0589] Biological evaluation

[0590] Test Example 1: Determination of the inhibitory activity of the compound of the present invention against recombinant human LRRK2 protein.

[0591] (I) Biochemical determination of compounds

[0592] • Experimental objective:

[0593] The energy signal transfer (520 nM / 485 nM fluorescence signal ratio) generated after the phosphate group of phosphorylated Fluorescein-ERM(LRRKtide)peptide binds to LanthaScreen.Tb-pERM(pLRRKtide)Antibody antibody was detected by homogeneous time-resolved fluorescence. The LRRK2 kinase inhibition IC50 value of the test compound was calculated.

[0594] Experimental materials:

[0595] 1. Reaction solution:

[0596] LRRK2:Invitrogen-PR8604B

[0597] Fluorescein-ERM(LRRKtide)peptide:Invitrogen-PV4901

[0598] ATP: Sigma-A7699

[0599] Tb-anti-pLRRKtide antibody:Invitrogen-PV4900

[0600] 2. Final concentration of LRRK2

[0601] LRRK2:2nM

[0602] Fluorescein-ERM(LRRKtide)peptide:400nM

[0603] ATP: 38μM

[0604] Tb-anti-pLRRKtide antibody:0.25nM

[0605] • Experimental procedure:

[0606] 1. The compounds were diluted using Echo, with final concentrations ranging from 10 μM to 0.17 nM;

[0607] 2. Add 5 μL of the protease and peptide mixture (containing the compound) to the wells of the detection plate;

[0608] 3. Centrifuge at 1000 rpm for about 15 seconds, then incubate at 23°C for 15 minutes;

[0609] 4. Add 5 μL of ATP solution per well to start the reaction;

[0610] 5. Centrifuge the detection plate at 1000 rpm for approximately 15 seconds, and then seal it with a sealing film;

[0611] Incubate at 6.23℃ for 120 minutes;

[0612] 7. Add 10 μL / well of antibody detection solution to stop the reaction;

[0613] 8. Separate the detection plate at 1000 rpm for about 15 seconds, then incubate at 23°C for 60 minutes;

[0614] 9. Read the detection board in Envision.

[0615] 10. The relative enzyme activity inhibition relative to the DMSO blank was calculated by the signal ratio, and the IC50 value was calculated by fitting the curve using the XLfit5 software.

[0616] • Experimental results:

[0617] Table 1 Results of LRRK2 kinase inhibitory activity assay

[0618]

[0619]

[0620] This experiment shows that these compounds have good LRRK2 enzyme inhibitory activity.

[0621] (II) Pharmacokinetic assays

[0622] • Experimental objective:

[0623] In vivo pharmacokinetics of the compound in C57BL / 6 mice: brain tissue and plasma drug concentration ratio (B / P ratio)

[0624] Experimental materials:

[0625] C57BL / 6 mice (male, 8 weeks old, weighing 25g-30g)

[0626] • Experimental procedure:

[0627] 1. The candidate compound was prepared into a 0.6 mg / mL solution and administered to mice via a single intravenous injection. The solvent was 5% dimethyl sulfoxide / 10% solubilitol / 85% physiological saline. Male C57BL / 6 mice were used in the experiment. The dosage was 3 mg / kg, and the administration volume was 5 mL / kg. Blood and whole brain tissue samples were collected at 0.25, 1, 4, and 24 hours after administration.

[0628] 2. Brain tissue was homogenized with 3x 0.9% NaCl and the homogenate was stored at -80°C for subsequent LC-MS / MS analysis.

[0629] 3. Within half an hour of blood sample collection, centrifuge at 4000 rpm for 15 minutes to separate the supernatant and obtain plasma. Store the plasma sample in polypropylene tubes.

[0630] 4. Before LC-MS / MS analysis, brain homogenate and plasma were added to acetonitrile solution containing internal standard to precipitate proteins. After thorough mixing and centrifugation, the supernatant was injected into the plasma. LC-MS / MS was used to quantitatively analyze the brain and plasma drug concentrations, and pharmacokinetic parameters such as peak concentration (Cmax), half-life (T1 / 2), time to peak concentration (Tmax), drug concentration in different tissues at different time points (AUC0-last), and the ratio of drug concentration in brain tissue to plasma (B / P) were calculated. Since Cpd-11 is a prodrug of Cpd-10, the pharmacokinetics of Cpd-11 were assessed by detecting the concentration of Cpd-10 in mice.

[0631] • Experimental results:

[0632] Table 2: Results of in vivo pharmacokinetic assays

[0633]

[0634] This experiment shows that these compounds have good brain distribution characteristics in mice.

[0635] The embodiments of the technical solution of the present invention have been described above by way of example. It should be understood that the protection scope of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the protection scope of the claims of this application.

Claims

1. A compound of Formula I, its racemate, isotope label, or pharmaceutically acceptable salt: in, R1 is selected from unsubstituted or arbitrarily assigned to one, two or more R1s. 11 The following groups are substituted: NH2; each R 11 Same or different, selected independently from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl, diC 1-6 alkyl-phosphoryl-phenyl; or R1 is selected from unsubstituted or optionally dominated by one C 1-6 Alkyl-substituted phenyl; R2 is selected from halogenated C 1-6 Alkyl groups, halogens; R3 is H; Alternatively, R2, R3, and the atoms they are bonded to form... X is selected from N; R4 is selected from H, F, Cl, Br, I, CN, C 1-6 Alkyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyloxy, halogenated C 1-6 Alkoxy, deuterated C 1-6 Alkyl group; m is selected from 1, 2, 3, 4, 5; Ring A is selected from C6 aryl and 5-membered heteroaryl groups; E is selected from chemical bonds or R E1 Selected from fluorine, chlorine, bromine, and iodine; G is selected from chemical bonds and C6 aryl groups; n is selected from 1 or 2; R5, R6, R7, and R8 may be the same or different, and are independently selected from H and C. 1-6 Alkyl, NH2 or NH2-C 1-6 Alkyl group; and at least one of R5, R6, R7, and R8 is NH2; Y is selected from OR a 、N(R b (R) c ); R a Selected from H, C 1-6 alkyl or R b R c Same or different, selected independently from C 1-6 alkyl.

2. The compound, its racemate, isotope label, or pharmaceutically acceptable salt according to claim 1, characterized in that, R1 is selected from methylamino, ethylamino, propylamino, isopropylamino, cyclopropylamino, cyclobutylamino, difluoroethylamino, trifluoroethylamino, tolyl, and dimethylphosphorylphenylamino. R2 is selected from trifluoromethyl, F, Cl, Br; Alternatively, R2, R3, and the atoms they are bonded to form...

3. The compound according to claim 1, its racemate, isotope label, or pharmaceutically acceptable salt, characterized in that, R4 is selected from H, F, Cl, CN, methyl, methoxy, ethoxy, cyclopropyl, cyclopropyloxy, difluoromethoxy, trifluoromethoxy, trideuterated methoxy, and ethynyl. And / or, R5, R6, R7, and R8 may be the same or different, and are independently selected from H, methyl, NH2, and NH2-methyl; and at least one of R5, R6, R7, and R8 is NH2 or NH2-methyl.

4. The compound according to claim 1, its racemate, isotope label, or pharmaceutically acceptable salt, characterized in that, When R5, R6, and R7 are all H, R8 is NH2; when R5, R7, and R8 are all H, R6 is NH2 or NH2-methyl; when R5 and R6 are both H, R7 is methyl and R8 is NH2; when R5 and R7 are both H, R6 is methyl or ethyl and R8 is NH2; and / or, when R8 is NH2, it is an S configuration; 5. The compound according to claim 1, its racemate, isotope label, or pharmaceutically acceptable salt, characterized in that, Ring A is selected from phenyl or pyrazolyl groups; E is selected from chemical bonds or When n is 1, R E1 It is an R configuration; Y is selected from OH, methylamino, dimethylamino, isopropyloxy, tert-butyloxy, heptayloxy, or...

6. The compound according to claim 5, its racemate, isotope label, or pharmaceutically acceptable salt, characterized in that, E is selected from chemical bonds, 7. The compound according to claim 6, its racemate, isotope label, or pharmaceutically acceptable salt, characterized in that, Selected from 8. The compound according to claim 1, its racemate, isotope label, or pharmaceutically acceptable salt, characterized in that, The compound shown in Formula I is selected from the structure shown in Formula II or Formula III: R1, R2, R3, R4, R5, R6, R7, R8, E, Y, and m independently have the definitions described in claim 1.

9. The compound according to claim 1, its racemate, isotope label, or pharmaceutically acceptable salt, characterized in that, The compound shown in Formula I is selected from the structure shown in Formula I-1, I-2, I-3 or I-4: R1, R4, R5, R6, G, and Y each have the definition described in claim 1.

10. The following compounds, their racemates, isotopic labels, or pharmaceutically acceptable salts:

11. A method for preparing the compound, its racemate, isotope label, or pharmaceutically acceptable salt according to any one of claims 1-10, characterized in that, Includes either Option 1 or Option 2: Option 1: Deprotection of compound I-1 yields the compound shown in Formula I; Option 2: Deprotection of compound I-2 yields the compound shown in Formula I; Wherein, R1, R2, R3, R4, R5, R6, R7, R8, m, A, E, X, and Y independently have the definition as described in any one of claims 1-10, and PG is selected from amino protecting groups.

12. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds of any one of claims 1-10, a racemic mixture thereof, an isotopic label thereof, or a pharmaceutically acceptable salt thereof.

13. The use of the compound of any one of claims 1-10, its racemate, isotopic label, or pharmaceutically acceptable salt, or the pharmaceutical composition of claim 12, in the preparation of an LRRK2 inhibitor.

14. The use of the compound of any one of claims 1-10, its racemate, isotopic label, or pharmaceutically acceptable salt, or the pharmaceutical composition of claim 12, in the preparation of a medicament for the prevention or treatment of diseases or symptoms mediated by LRRK2.

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