Pyrimidinamine NUAK inhibitor, and preparation method therefor and use thereof

Abstract

Provided in the present application is a pyrimidinamine compound, which is characterized in that the pyrimidinamine compound is a compound as shown in formula I, or a stereoisomer, tautomer, isotopic derivative, hydrate, solvate, prodrug and pharmaceutically acceptable salt thereof. The pyrimidinamine compound of the present application has an excellent NUAK1 / NUAK2 kinase inhibitory activity, and can be used for preventing or treating the following diseases by inhibiting NUAK1 / NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrotic diseases and skin fibrotic diseases.

Description

A pyrimidineamine NUAK inhibitor and its preparation method and use

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application No. 202310860161.7 filed on July 13, 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention belongs to the field of small molecule compounds, specifically to a pyrimidineamine-based NUAK inhibitor, its preparation method, and use. By inhibiting NUAK1 / NUAK2, the compound can be used to prevent or treat the following diseases: neuropsychiatric disorders, metabolic diseases, tumors, visceral fibrosis, skin fibrosis, or to alleviate scarring after trauma and surgery. Background Art

[0004] Protein kinases are a group of important functional proteins involved in regulating cellular metabolism, polarity, growth, division, and differentiation. The human genome encodes over 500 protein kinases, which transfer phosphate groups from ATP (adenosine triphosphate) to specific serine, threonine, or tyrosine residues on substrate proteins, thereby phosphorylating them. Most protein kinases are serine / threonine kinases, while there are fewer than 100 tyrosine kinases. Adenosine monophosphate-activated protein kinase (AMPK) belongs to the serine / threonine kinases (STKs) and is a key regulator of cellular energy homeostasis in mammals. It regulates glucose and lipid metabolism, cell proliferation, and cell polarity by sensing changes in the intracellular AMP (adenosine monophosphate) / ATP or ADP (adenosine diphosphate) / ATP ratios in response to metabolic stress (such as hypoxia and heat shock), thus regulating glucose and lipid metabolism, cell proliferation, and cell polarity. It is considered a metabolic sensor protein. AMPK is highly conserved in evolution and is a heterotrimeric protein composed of an α subunit containing a kinase domain (KD), and β and γ subunits that regulate kinase activity. The γ subunit contains four CBS functional domains (Cystathionine-β-synthase (CBS) domains), which are responsible for detecting changes in the AMP / ATP and ADP / ATP ratios in the cell (Hardie DG, Trends in Cell Biology. 2016, 26: 190).

[0005] AMPK dysfunction can lead to a variety of diseases such as obesity, diabetes, inflammatory diseases and tumors, so the body's regulation of AMPK function is very critical. First, the activation of AMPK requires upstream kinases to phosphorylate the threonine (T172) at position 172 of its α subunit. Currently, there are three known upstream kinases of AMPK: liver kinase B1 (LKB1), calcium ion / calmodulin-dependent protein kinase 2 (Ca2+), and AMPK-dependent protein kinase 2 (CMP-2). 2+ / calmodulin-dependent PK kinase 2, CaMKK2), and transforming growth factor-β-activated kinase 1 (TAK1). Corresponding to the activation of AMPK by upstream kinases through phosphorylation of T172 site, three protein phosphatases are known to inhibit AMPK activity by removing the phosphate group on T172, including protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), and magnesium / manganese-dependent protein phosphatase 1E (Mg / Mn-dependent protein phosphatase 1E). 2+ - / Mn 2+ -dependent protein phosphatase 1E, PPM1E). When cells are in a low-energy state (high AMP / ATP or ADP / ATP ratio), the α subunit KD and γ subunit of AMPK are tightly cross-linked, and the β subunit is myristoylated, ensuring that the protein phosphatase cannot access the T172 site and remains activated. When cells are in a high-energy state, the KD and γ subunits are loosened, exposing T172 to the phosphatase and causing AMPK inactivation (Steinberg GR, Nature Reviews Drug Discovery. 2019, 18:527).

[0006] In addition to the upstream kinases and protein phosphatases that activate and inactivate AMPK, a class of AMPK-related kinases (ARKs) also participate in regulating AMPK function. Currently, there are 12 ARKs (BRSK1, BRSK2, NUAK1, NUAK2, QIK, QSK, SIK, MARK1, MARK2, MARK3, MARK4, and MELK). All are serine / threonine protein kinases. The kinase domains of ARKs share high homology with the α subunit of AMPK. With the exception of MELK, all ARKs can be activated by LKB1. Their kinase activation phosphorylation site is equivalent to T172 of AMPK. Functionally, all ARKs participate in the regulation of cellular metabolism, proliferation, and polarity. However, unlike AMPK, ARKs cannot be directly regulated by the intracellular AMP / ATP ratio because they possess no regulatory subunits (Bright NJ, Acta Physiologica. 2009, 196:15).

[0007] ARKs are further divided into several ARK subfamilies due to differences in protein structure and functional characteristics. The NUAK (Nu(novel) and AMPK-related kinase) ARK subfamily contains two members, NUAK1 (originally named ARK5) and NUAK2 (originally named sucrose nonfermenting-like / AMPK-related kinase, SNARK). The amino acid sequences of the two members are approximately 55% homologous. Based on the amino acid sequences, the estimated molecular weights of NUAK1 and NUAK2 are 76 and 69 kDa, respectively. Their protein structures are very similar, with a kinase domain at the amino terminus and a ubiquitin-associated domain at the carboxyl terminus. It is currently unclear whether NUAK is also a heterotrimeric structure like AMPKs. NUAK1 and NUAK2 are expressed in most tissues, with NUAK1 expression significantly higher in organs and tissues such as the brain, skin, muscle, upper digestive tract, and endocrine system than in other parts of the body. NUAK2 expression is highest in the digestive tract, female reproductive system, skin, bone, brain, and endocrine system, with NUAK2 expression showing greater tissue specificity. It is worth noting that NUAK1 and other ARKs and AMPK proteins are primarily distributed in the cytoplasm, while NUAK2 is primarily distributed in the nucleus. There are also indications that NUAK2 acts as a transcriptional regulator in the expression of genes associated with metabolic stress (Sun X, J of Molecular Endocrinology. 2013, 51: R15).

[0008] As serine / threonine protein kinases, NUAK1 and NUAK2 phosphorylate a variety of protein substrates, including proteins involved in cell signaling, metabolism, cell proliferation, apoptosis, autophagy, and cytoskeletal organization. NUAK1 is known to phosphorylate AMPK, LATS1 / 2, the p53 tumor suppressor protein, and myosin phosphatase target subunit 1 (Mypt1), which regulates actin cytoskeletal organization. NUKA2 phosphorylates the Hedghog signaling pathway transcription factor Gli3, FoxO1, kinesin light chain 1 (KLC1), and Rho GDP dissociation inhibitor α (Rho GDIα).

[0009] The functional regulation of NUAK1 and NUAK2 involves multiple intracellular signaling systems. Besides being activated by LKB1 phosphorylation (T211, equivalent to T172 in AMPK) and calcium / PKC activation, NUAK1 is the only member of the AMPK-related protein family that can be activated by Akt. Increased NUAK1 activity is also observed in response to activation of growth factor signaling pathways, such as insulin-like growth factor 1 (IGF1). NUAK1 activity is also enhanced during skeletal muscle contraction. Both NUAK1 and NUAK2 are activated by LKB1 (T211 / NUAK1, T208 / NUAK2, equivalent to T172 in AMPK), but NUAK2 can also undergo autophosphorylation. NUAK2 interacts with ubiquitin-specific protease 9 (USP9X), a deubiquitinating enzyme on the X chromosome, to maintain NUAK2 activity. In different cells, stimuli such as low osmotic pressure, DNA damage, oxidation, and malnutrition can lead to the activation of NUAK2. NUAK2 activation is also seen in the contraction of skeletal muscle cells (Brooks D, Data in Brief. 2022, 43:108482).

[0010] Numerous studies have shown that NUAK plays a crucial role in the pathogenesis of metabolic diseases, tumors, neurodegenerative diseases, and fibrotic disorders. Homozygous knockout of NUAK1 and NUAK2 in mice generally leads to embryonic lethality. NUAK1 heterozygous deletion in mice leads to dysplasia of the body and neural tissues (such as the cerebral cortex and peripheral neurons). Heterozygous mutations in human NUAK1 are associated with autism spectrum disorders (ASD), cognitive deficits, attention deficit / hyperactivity disorder (AD / HD), and schizophrenia. NUAK2 gene deletion in humans leads to anencephaly, a severe neural tube defect that causes fetal developmental defects. Its mechanism is related to the functional loss of YAP (Bonnard C, J Exp Med. 2020, 217:e20191561). Skeletal muscle cell-specific knockout of the NUAK1 gene can prevent subclinical diabetes induced by a high-fat diet; the phenotype of NUAK2 heterozygous deletion mice is similar to a series of clinical manifestations of human type 2 diabetes accompanied by obesity, which may be related to an imbalance in the cellular autophagy mechanism (Bennison SA, Cellular Signaling. 2022, 100: 110472; Blazejewski SM, Scientific Reports. 2011, 11: 8156).

[0011] NUAK1 activation by Akt and other protein kinases promotes tumor cell survival in energy-deficient environments and protects tumor cells from apoptosis. NUAK2 also inhibits TNFα- and CD95-induced apoptosis through similar mechanisms. Furthermore, activated NUAK1 promotes tumor cell invasion and metastasis by upregulating matrix metalloproteinases (MMPs), while NUAK2 participates in tumor metastasis by enhancing tumor cell activity (Hou X, Oncogen. 201, 30:2933; Chen Y, Cell Death and Disease. 2020, 11:712; Molina E, Cells. 10:2760; Humbert N, The EMBO Journal. 2010, 29:376). Numerous lines of evidence suggest that NUAK1 and NUAK2 play a role in tumor formation and metastasis. Compared to NUAK1, NUAK2 has a stronger role in promoting tumor formation and metastasis. Therefore, NUAK2 inhibitors are more likely to become new targeted anti-tumor drugs than NUAK1 inhibitors.

[0012] Recent studies have shown that NUAK plays a key role in the development of tissue fibrosis through interaction with the transforming growth factor-β (TGF-β) signaling pathway. First, TGF-β upregulates NUAK1 and NUAK2 gene transcription in various epithelial cells, such as keratinocytes and human dermal fibroblasts, while inactivation of MAPK (ERK1 / 2 and p38) signaling inhibits TGF-β-dependent NUAK2 expression. Furthermore, NUAK2 inhibits the intracellular degradation of SMAD3 by binding to the crosslinking domain and MH2 domain of the TGF-β intracellular signaling mediator SMAD3. A similar mechanism also exists between NUAK2 and TβRI. TGF-β-induced gene expression of pro-fibrotic molecules such as fibronectin (FN), plasminogen activator inhibitor 1 (PAI1), and tissue inhibitor of metalloproteinase-1 (TIMP1) depends on the presence of NUAK2, which shows that NUAK2 promotes the occurrence of fibrosis. Studies have also shown that NUAK1 promotes renal, lung, and liver fibrosis by upregulating the TGF-β and YAP signaling pathways. In animal experiments, inhibiting NUAK1 can reduce scars caused by new trauma and old scar tissue (Gill MK, Nature Communications. 2018.9:3510). Interestingly, studies have found that the expression of the gene encoding FN is upregulated in keratinocytes with NUAK1 knockout genes. NUAK1 may affect TGF-β signaling through a negative feedback mechanism, thereby inhibiting fibrosis (van de Vis RAJ, Cancers. 2021, 13:3377). In general, inhibiting NUAK may effectively inhibit the fibrosis process of participating tissues and organs.

[0013] Because kinases have long been ideal targets for small molecule drugs based on their structure and function, and NUAK1 and NUAK2 are involved in the pathogenesis of various diseases, several NUAK inhibitors are currently in early stages of development, but their efficacy and safety remain uncertain (Banerjee S, The Biochemical Journal. 2014, 457:215). Therefore, the development of NUAK-selective inhibitors and NUAK1 / 2 dual-target inhibitors has the potential to provide innovative treatments for neuropsychiatric disorders (such as Parkinson's disease and Alzheimer's disease), metabolic diseases (such as diabetes, hyperlipidemia, and obesity), tumors (such as liver cancer, leukemia, lymphoma, and tumor metastasis), visceral fibrotic diseases (such as cirrhosis, renal fibrosis, pulmonary fibrosis, and myocardial sequelae), and skin fibrotic diseases (such as scleroderma, keloids, and hypertrophic scars, or simply to alleviate scarring after trauma and surgery). The development of a new generation of NUAK inhibitors has enormous potential for clinical application.

[0014] Summary of the Invention

[0015] The purpose of the present invention is to obtain effective NUAK1 / NUAK2 inhibitors that can be used to prepare drugs for preventing or treating the following diseases: neuropsychiatric diseases (such as Parkinson's disease, Alzheimer's disease), metabolic diseases (such as diabetes, hyperlipidemia, obesity), tumors (such as liver cancer, leukemia, lymphoma, tumor metastasis), visceral fibrosis diseases (such as cirrhosis, renal fibrosis, pulmonary fibrosis, and sequelae of myocarditis) and skin fibrosis diseases (such as scleroderma, keloids, hypertrophic scars, or simply for alleviating traumatic and post-operative scars).

[0016] To achieve the above objectives, in one aspect, the present invention provides a pyrimidineamine compound, which is a compound represented by Formula I shown below, or its stereoisomers, geometric isomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs, and pharmaceutically acceptable salts:

[0017] Wherein, X is a direct bond, -O-, -S-, -NR7- or -CR8R9-, and Y is -CO- or -S(O)2-;

[0018] A is an optionally substituted 6-10 membered aryl or 5-10 membered heteroaryl, B is an optionally substituted 3-10 membered cycloalkyl or 3-10 membered heterocycloalkyl, and the substituents on A or B are selected from one or more of halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino;

[0019] R1, R2, R6, R7, R8, and R9 are each independently H, or an optionally substituted C1-8 alkyl group;

[0020] R3 and R4 are each independently halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, or optionally substituted C1-8 alkylamino;

[0021] R5 is an optionally substituted 3-10 membered cycloalkyl, an optionally substituted 3-10 membered heterocycloalkyl, wherein the substituent on the cycloalkyl or heterocycloalkyl is selected from one or more of halogen, amino, hydroxy, nitro, cyano, thiol, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino;

[0022] The substituents on the C1-8 alkyl group are selected from one or more of halogen, amino, hydroxy, nitro, cyano, and mercapto;

[0023] n is 0, 1 or 2, and m is 0, 1, 2, 3, 4 or 5.

[0024] In one group of embodiments, R5 is an optionally substituted 3-10 membered cycloalkyl, preferably an optionally substituted 3-6 membered cycloalkyl, preferably a halogen substituted 3-6 membered cycloalkyl, more preferably R5 is a fluorine substituted 3-6 membered cycloalkyl.

[0025] Furthermore, R5 is Preferably R5 is

[0026] In one group of embodiments, R6 is methyl.

[0027] In one group of embodiments, R2 is H.

[0028] In one group of embodiments, B is an optionally substituted 3-6 membered heterocycloalkyl, preferably B is an optionally substituted 5-6 membered heterocycloalkyl, more preferably B is an optionally substituted piperidinyl or an optionally substituted piperazinyl.

[0029] In one group of embodiments, the compound represented by Formula I is Formula I-1 or Formula I-2,

[0030] R 10 Selected from halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino;

[0031] R 11 is H or optionally substituted C1-8 alkyl;

[0032] The substituents on the C1-8 alkyl group are selected from one or more of halogen, amino, hydroxy, nitro, cyano, and mercapto;

[0033] q is 0, 1, 2, 3 or 4.

[0034] Furthermore, R 11 is H or methyl.

[0035] In one group of embodiments, A is optionally substituted phenyl or optionally substituted 5-membered heteroaryl.

[0036] Furthermore, A is one of the following groups which are optionally substituted:

[0037] Preferably, it is connected to the amino group at the top and to the B group at the bottom.

[0038] In one group of embodiments, R3 is chloro, preferably substituted in the para position relative to the amino group.

[0039] In one group of embodiments, the substituent on the A ring is methoxy, preferably substituted at the ortho position to the amino group.

[0040] In one embodiment, the compound represented by formula I is one of the following compounds:

[0041] In one group of embodiments, the pharmaceutically acceptable salt is a formate salt.

[0042] In one set of embodiments, the isotopic derivative is a deuterated derivative.

[0043] On the other hand, the present application also provides a method for preparing the pyrimidineamine compound, which comprises the following steps: preparing a compound of formula I from a compound of formula II and a compound of formula III;

[0044] or

[0045] Prepare a compound of formula V from compounds of formula IV and formula II, and then prepare a compound of formula I from the compound of formula V;

[0046] Optionally, the preparation process includes necessary protection and deprotection steps;

[0047] Wherein, Z is a leaving group, preferably Z is halogen, more preferably Z is Cl, and the definitions of the other groups are as defined above.

[0048] In one embodiment, the amino group or imino group on the B ring in formula II is first protected, coupled with the compound of formula III, and then the protecting group is removed to prepare the compound of formula I.

[0049] In one group of embodiments, the amino or imino group attached to R2 in the compound of formula IV is first protected, and the amino or imino group on the B ring in formula II is optionally protected. The protected compound of formula IV is then coupled with the optionally protected compound of formula II, and the protecting group on the amino or imino group attached to R2 is removed to prepare the compound of formula V. The compound of formula V is then reacted with R5-Y-OH, and the protecting group on the B ring is optionally removed to prepare the compound of formula I.

[0050] The present application provides a pharmaceutical composition, which uses the pyrimidineamine compound described above in the present application as an active ingredient; the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0051] The present application provides the use of the pyrimidineamine compounds for preparing NUAK1 or NUAK2 inhibitors.

[0052] The present application provides the use of the pyrimidineamine compound for preparing a drug, characterized in that the compound is used to prevent or treat the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases.

[0053] In one group of embodiments, the disease is Parkinson's disease, Alzheimer's disease, diabetes, hyperlipidemia, obesity, liver cancer, leukemia, lymphoma, tumor metastasis, cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or is used alone to reduce scars after trauma and surgery.

[0054] The beneficial effects of the present invention are:

[0055] The present invention provides a class of pyrimidineamine compounds. In vitro kinase activity inhibition assays demonstrate that the compounds exhibit excellent inhibitory activity against NUAK1 / NUAK2 kinases. These compounds can be used to prevent or treat the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric disorders, metabolic diseases, tumors, visceral fibrosis, and skin fibrosis. DETAILED DESCRIPTION

[0056] The following is a detailed description of the specific embodiments of the present invention. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention and are not intended to limit the present invention.

[0057] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0058] Before describing the present invention in detail, it should be understood that the terminology used herein is intended only to describe specific embodiments and is not intended to limit the scope of the invention, which is defined solely by the appended claims. In order to more fully understand the invention described herein, the following terms are used and their definitions are as follows. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as understood by one of ordinary skill in the art to which the invention belongs.

[0059] definition

[0060] Unless otherwise specified, the following terms used in the present invention have the following definitions.

[0061] Features illustrated or described as part of one embodiment or group of embodiments can be used on another embodiment or group of embodiments to yield a still further embodiment.

[0062] In the present invention, the ring group It means there can be n R3, m R4, q R 10 Attachment is at any possible position on the ring group.

[0063] In the present invention, some substituents Indicates the connection site.

[0064] Unless otherwise indicated, the term "optionally substituted" means that the hydrogen on the substituted group is not replaced or one or more substitutable sites of the substituent group are independently replaced by substituents, and the substituents are independently selected from one or more of deuterium, halogen, amino, hydroxyl, nitro, cyano, mercapto, oxo, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino; the substituent on the C1-8 alkyl group is selected from one or more of deuterium, halogen, amino, hydroxyl, nitro, cyano, and mercapto; when the substituent is selected from "oxo", it means that two hydrogen atoms at the same substitution position are replaced by oxygen atoms.

[0065] The term "aryl" refers to a monocyclic or bicyclic aromatic carbon ring system containing 6 to 10 carbon atoms. Examples of aryl groups include phenyl and naphthyl.

[0066] The term "heteroaryl" refers to an aromatic monocyclic or polycyclic ring system having a 5-10 membered structure, or preferably a 5-8 membered structure, more preferably a 5-6 membered structure, wherein 1, 2, 3, 4 or more ring atoms are heteroatoms and the remaining atoms are carbon, the heteroatoms are independently selected from O, N or S, and the number of heteroatoms is preferably 1, 2, 3 or 4. The heteroaryl group may be a 5-6 membered aromatic monocyclic ring containing 1-2 atoms selected from N or S, or a 5-6 membered aromatic monocyclic ring containing 1-2 N atoms. Examples of heteroaryl groups include, but are not limited to, furyl, thienyl, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiodiazolyl, triazinyl, phthalazinyl, quinolyl, isoquinolyl, pteridinyl, purinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, benzothiophenyl, benzopyridinyl, benzopyrimidinyl, benzo pyrazinyl, benzimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridinyl, imidazo[1,2-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazolo[1,5-a]pyridinyl, etc.

[0067] The term "cycloalkyl" refers to a saturated monocyclic, bicyclic or tricyclic ring system containing 3-10 carbon atoms, wherein the monocyclic, bicyclic or tricyclic ring does not contain an aromatic ring. Bicyclic groups include bridged ring groups, spirocyclic groups, and annular groups. Preferably, the cycloalkyl group contains 3-10 carbon atoms (C3-10 cycloalkyl), and more preferably, the cycloalkyl group contains 3-8 carbon atoms (C3-8 cycloalkyl), 3-6 carbon atoms (C3-6 cycloalkyl), 4-6 carbon atoms (C4-6 cycloalkyl), and 5-6 carbon atoms (C5-6 cycloalkyl). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, etc.

[0068] The term "heterocycloalkyl" refers to a saturated monocyclic, bicyclic, or polycyclic hydrocarbon group, preferably containing 3-10 ring atoms, of which 1, 2, 3, or more ring atoms are selected from N, O, or S, and the remaining ring atoms are C, including bridged ring groups, spiro ring groups, and annular ring groups. Preferably, it contains 3-8 ring atoms (3-8 membered heterocycloalkyl), or 3-6 ring atoms (3-6 membered heterocycloalkyl), or 4-6 ring atoms (4-6 membered heterocycloalkyl), or 5-6 ring atoms (5-6 membered heterocycloalkyl). The number of heteroatoms is preferably 1-4, more preferably 1-3 (i.e., 1, 2, or 3). The heterocycloalkyl group may be a 5-6 membered monocyclic heterocycloalkyl group containing 1-2 nitrogen atoms. Examples of heterocycloalkyl groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, pyranyl, aziridine, oxirane, thiirane, azetidinyl, oxetanyl, thietanyl, oxane, morpholinyl, thiomorpholinyl, dioxanyl, dithioxanyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, imidazolinidine, and the like.

[0069] The term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, preferably a straight or branched chain group containing 1 to 8 carbon atoms (C1-8 alkyl), wherein the number of carbon atoms is between 1 and 8, specifically 1, 2, 3, 4, 5, 6, 7, or 8, and more preferably a C1-6 alkyl group containing 1 to 6 carbon atoms, specifically 1, 2, 3, 4, 5, or 6. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0070] The terms "alkyloxy," "alkylthio," and "alkylamino" refer to -O-alkyl, -S-alkyl, -NH-alkyl, or dialkylamino, respectively, wherein alkyl is as defined above. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, and tert-butoxy; methylthio, ethylthio, propylthio, isopropylthio, butylthio, 1-methylpropylthio, 2-methylpropylthio, and tert-butylthio; and methylamino, ethylamino, propylamino, dimethylamino, diethylamino, dipropylamino, and methylethylamino.

[0071] The term "halogen" refers to F, Cl, Br, I.

[0072] The active compounds of the present invention are interpreted to include the compounds and their stereoisomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts. The stereoisomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs, isotopic derivatives, or pharmaceutically acceptable salts are obtained by conventional techniques in the art and exert the same or similar effects in vivo and in vitro through substantially the same mechanism of action as the compounds.

[0073] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including configurational isomers and conformational isomers, wherein configurational isomers further include geometric isomers (or cis-trans isomers) and optical isomers (including enantiomers and diastereomers). Geometric isomers may exist in the present compound. Optical isomers refer to substances with identical molecular structures and similar physicochemical properties but different optical rotations. The compounds of the present invention may contain asymmetrically substituted carbon atoms in the R or S configuration, wherein the terms "R" and "S" are defined as in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem. (1976) 45, 13-10. Compounds with asymmetrically substituted carbon atoms (having equal numbers of R and S configurations) are racemic at those carbon atoms. Atoms having an excess of one configuration (relative to the other) result in that configuration being present in a higher number, preferably in an excess of about 85% to 90%, more preferably in an excess of about 95% to 99%, and even more preferably in an excess of greater than about 99%. Accordingly, the present invention includes racemic mixtures, relative and absolute optical isomers, and mixtures of relative and absolute optical isomers.

[0074] The term "tautomer" refers to structural isomers with different energies that can be interconverted through a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of the tautomers can be reached. For example, proton tautomers (also known as prototropic tautomers) include interconversions that occur through proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions that occur through the reorganization of some of the bonding electrons.

[0075] The term "isotopic derivative" means that the compounds of the present invention may exist in an isotopically-tagged or enriched form containing one or more atoms having an atomic mass or mass number that is different from the atomic mass or mass number of the atom found in the largest amount in nature. Isotopes may be radioactive or non-radioactive. Isotopes of atoms such as hydrogen, carbon, phosphorus, sulfur, fluorine, chlorine, and iodine include, but are not limited to: 2 H,3 H, 13 C, 14 C, 15 N, 18 O, 32 P, 35 S, 18 F, 36 Cl and 125 I. Compounds containing other isotopes of these and / or other atoms are within the scope of the present invention. Isotopically labeled compounds of the present invention can be prepared by general methods well known to those skilled in the art.

[0076] The term "hydrate" refers to an association of one or more water molecules with a compound of the present invention.

[0077] The term "solvate" refers to an association between one or more solvent molecules and a compound of the present invention.

[0078] The term "prodrug" is a derivative of an active drug that is designed to improve some defined, undesirable physical or biological property. Physical properties are usually related to solubility (too high or insufficient lipid or water solubility) or stability, while problematic biological properties include rapid metabolism or poor bioavailability, which themselves may be related to physicochemical properties.

[0079] The term "pharmaceutically acceptable salt" refers to salts that are suitable for use in contact with mammalian tissues, particularly human tissues, without excessive toxicity, irritation, or allergic reaction, and are commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment. If the compound is basic, pharmaceutically acceptable salts include salts prepared from inorganic acids and also include salts prepared from organic acids. If the compound is acidic, pharmaceutically acceptable salts include salts prepared from inorganic bases and / or organic bases.

[0080] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant government regulatory authorities as acceptable for use by humans or livestock.

[0081] As used herein, the term "treatment" refers to any administration of a therapeutic agent according to a therapeutic regimen that achieves the desired effect, i.e., partially or completely alleviates, improves, alleviates, inhibits, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms or characteristics of a particular disease, disorder, and / or condition; in some embodiments, the administration of a therapeutic agent according to a therapeutic regimen is associated with the achievement of the desired effect. Such treatment may be directed to subjects who do not exhibit the relevant disease, disorder, and / or condition and / or to subjects who only exhibit early signs of the disease, disorder, and / or condition. Alternatively or in addition, such treatment may be directed to subjects who exhibit one or more determined signs of the relevant disease, disorder, and / or condition. In some embodiments, treatment may be directed to subjects who have been diagnosed with the relevant disease, disorder, and / or condition. In some embodiments, treatment may be directed to subjects who are known to have one or more susceptibility factors that are statistically associated with an increased risk of developing the relevant disease, disorder, and / or condition.

[0082] According to the present invention, the medicament prepared for the pharmaceutical use described herein may contain, in addition to the pyrimidineamine compound of the present invention as an active ingredient, another agent useful for preventing or treating related diseases as another active ingredient. When the medicament contains multiple active ingredients, each active ingredient may be administered simultaneously, sequentially, or separately at the physician's discretion.

[0083] Hereinafter, the effects of the specific compounds of the present invention will be described in detail with reference to Examples.

[0084] Example

[0085] Example 1 General method for the synthesis of compound 537 (TDM-181137)

[0086] Step 1: Compound 537c

[0087] tert-Butyl 1-(2-chloropyrimidin-4-yl)-4-methylpiperidin-4-carbamate

[0088] To a solution of compound 537a (300 mg, 2.01 mmol) and compound 537b (430.74 mg, 2.01 mmol) in acetonitrile (30 ml) was added potassium carbonate (555.60 mg, 4.02 mmol), and the reaction solution was stirred at 50° C. for 1 hour. After the reaction, the mixture was concentrated under reduced pressure, water (60 mL) was added to the residue, and the mixture was extracted with ethyl acetate (3*30 ml). The organic layers were combined, washed with brine (50 mL), and dried over sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 0-24 / 76) to obtain a colorless solid compound (compound 537c, 548.3 mg, 83.5% yield). LCMS [M+1] + =327.1.

[0089] Step 2: Compound 537d

[0090] 4-Chloropyrimidine-4-chloropyridin-4-amine

[0091] To a solution of compound 537c (548.3 mg, 1.68 mmol) in dichloromethane (22 mL) was added trifluoroacetic acid (2.49 mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated under reduced pressure, water (60 mL) was added to the residue, and the base was adjusted with potassium carbonate solution. Ethyl acetate (3*50 ml) was added for extraction, and the organic layers were combined, washed with brine (50 mL), and dried over sodium sulfate. The filtrate was concentrated under reduced pressure to obtain a brown compound (compound 537d, 337.2 mg, 88.5% yield), and the crude product was used directly in the subsequent step. LCMS [M+1] + =227.1.

[0092] Step 3: Compound 537f

[0093] N-(1-(2-chloropyrimidin-4-yl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0094] To a solution of compound 537d (337.2 mg, 1.49 mmol) in N,N-dimethylformamide (15 mL) were added triethylamine (0.62 mL, 4.46 mmol) and compound 537e (0.21 mL, 2.23 mmol), and the mixture was stirred at room temperature for 2 hours. After the reaction, the mixture was concentrated under reduced pressure, water (60 mL) was added to the residue, and the mixture was extracted with ethyl acetate (40 mL*3). The organic layers were combined, washed with brine (50 mL), and dried over sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0 to 44 / 56) to obtain a white solid compound (compound 537f, 231.8 mg, yield 47%). LCMS [M+1] + =331.1.

[0095] Step 4: Compound 535h

[0096] tert-Butyl 4-(4-(4-(4-cyclopropanesulfonamido)-4-methylpiperidin-1-yl)pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0097] To a solution of compound 537f (100 mg, 0.3 mmol) and compound 537g (96.6 mg, 0.36 mmol) in dioxane hydrochloride (5 mL) were added 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (69.89 mg, 0.12 mmol), palladium acetate (13.56 mg, 0.06 mmol), and cesium carbonate (195.49 mg, 0.6 mmol). The mixture was degassed under vacuum, replaced with argon, and the reaction solution was stirred at 100° C. for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane:dichloromethane containing 10% methanol = 0-60 / 40) to give a brown solid product (compound 537h, 106.4 mg, 63.7% yield). LCMS [M+1] + =561.2.

[0098] Step 5: Compound 535

[0099] N-(4-Methyl-1-(2-(1-(4-piperidinyl)-1H-pyrazol-4-amino)pyrimidin-4-ylpiperidin-4-yl)cyclopropanesulfonamide

[0100] To a solution A (dichloromethane:methanol = 10:1) of compound 537h (100 mg, 0.178 mmol) was added dioxane hydrochloride (0.89 mL), and the reaction solution was stirred at room temperature for 2 h. Upon completion of the reaction, the mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to yield a white solid compound (compound 537, 18.9 mg, 23% yield). LCMS [M+1] + =461.2.

[0101] 1 H NMR (400MHz, MeOD) δ7.93 (s, 1H), 7.66 (s, 2H), 6.58 (d, J = 7.3Hz, 1H), 4.59 (s,2H),3.91(s,1H),3.66(s,2H),3.57(d,J=12.7Hz,2H),3.23(t,J=10.8 Hz,2H),2.63(dq,J=7.9,4.9Hz,1H),2.33(s,4H),2.19(d,J=13.9Hz,2H), 1.65(t,J=10.6Hz,2H),1.52(s,3H),1.15–1.10(m,2H),1.09–1.03(m,2H).

[0102] Example 2: General method for synthesizing compound 563 (TDM-181163)

[0103] Step 1: Compound 563c

[0104] tert-Butyl (1-(2-chloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamate

[0105] Potassium carbonate (557 mg, 4.028 mmol) was added to a solution of compound 563a (300 mg, 2.014 mmol) and compound 563b (453 mg, 2.115 mmol) in acetonitrile (30 mL). The mixture was heated to 50°C and stirred overnight. The mixture was added to water (150 mL) and extracted with ethyl acetate (50 mL*3). The organic layers were combined, washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (ethyl acetate / petroleum ether = 0-45 / 55) to give a white solid product (compound 563c, 528.5 mg, 80.3% yield). LCMS [M+1] + =327.

[0106] Step 2: Compound 563e

[0107] Benzyl-4-(4-((4-(4-(tert-butoxycarbonyl)amino)-4-methylpiperidin-1-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0108] To a mixture of compound 563c (109 mg, 0.333 mmol), compound 563d (100 mg, 0.333 mmmol), palladium acetate (7.5 mg, 0.033 mmol), xantphos (38 mg, 0.067 mmol) and cesium carbonate (217 mg, 0.666 mmol) was added dioxane (5 mL). The mixture was degassed under vacuum, replaced with argon several times, then heated to 100 ° C and stirred for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane=0~62 / 38) to give a white solid product (compound 563e, 131.3 mg, 66.7% yield). LCMS[M+1] + =591.

[0109] Step 3: Compound 563f

[0110] Benzyl 4-(4-((4-(4-amino-4-methylpiperidin-1-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0111] To a solution of compound 563e (90 mg, 0.152 mmol) in dichloromethane (9 mL) was added trifluoroacetic acid (0.7 mL) at room temperature. The mixture was stirred for 1 hour. The mixture was then concentrated under reduced pressure, the residue was added with water and neutralized with aqueous sodium carbonate solution, extracted with ethyl acetate (20 mL*3), the organic layers were combined, washed with brine, and the filtrate was concentrated under reduced pressure to give a yellow solid product (compound 563f, 73.5 mg, 98.5% yield). LCMS [M+1] + =491.

[0112] Step 4: Compound 563h

[0113] Benzyl (S)-4-(4-((4-(4-(2,2-difluorocyclopropane-1-carboxamido)-4-methylpiperidin-1-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0114] To a solution of compound 563f (73.5 mg, 0.15 mmol) and N,N-diisopropylethylamine (29 mg, 0.225 mmol) in N,N-dimethylformamide (5 mL) was added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (85 mg, 0.22 mmol) and compound 563f (27.5 mg, 0.225 mmol) at room temperature. The mixture was stirred for 1 hour. The mixture was then added with water and extracted with ethyl acetate (30 mL*3). The organic layers were combined and washed with brine. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0-70 / 30) to give a yellow solid product (compound 563h, 80 mg, 61.1% yield). LCMS [M+1] + =595.

[0115] Step 5: Compound 563

[0116] (S)-2,2-Difluoro-N-(4-methyl-1-(2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)piperidin-4-ylcyclopropane-1-carboxamide

[0117] Compound 563h (70 mg, 0.118 mmol) was dissolved in HBr / CH3COOH (2 mL) at 0°C, and the mixture was stirred at 0°C for 30 minutes. The mixture was then concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to give a white solid product (compound 563, TDM-181163, 5.9 mg, 10.9% yield). LCMS [M+1] + =486.3.

[0118] 1 H NMR (400MHz, DMSO) δ8.84(s,1H),8.18(s,1H),7.99(s,1H),7.89(d,J=6.0Hz,1H),7.80(s, 1H),7.51(s,1H),6.19(d,J=6.1Hz,1H),4.39(dt,J=14.9,5.4Hz,1H),3.90(s,2H),3.36(d, J=12.8Hz,2H),3.26(s,3H),3.05–2.97(m,2H),2.67(ddd,J=14.7,10.9,8.0Hz,1H),2.13(d ,J=13.7Hz,4H),2.09–1.97(m,2H),1.90–1.72(m,2H),1.49(d,J=11.1Hz,2H),1.32(s,3H).

[0119] Example 3: General method for the synthesis of compound 564 (TDM-181164)

[0120] Step 1: Compound 564

[0121] (S)-2,2-Difluoro-N-(4-methyl-1-(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)piperidin-4-yl)cyclopropane-1-carboxamide

[0122] To a solution of compound 564a (30 mg, 0.079 mmol) and N,N-diisopropylethylamine (15.2 mg, 0.118 mmol) in N,N-dimethylformamide (3 mL) was added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (44.8 mg, 0.118 mmol) and compound 564b (19.2 mg, 0.157 mmol) at room temperature. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (100% methanol) and preparative HPLC (formic acid) to give a yellow solid product (compound 564, 6.2 mg, 16.2% yield). LCMS [M+1] + =486.3.

[0123] 1 H NMR (400MHz, DMSO) δ8.75 (s, 1H), 8.16 (s, 2H), 7.97 (s, 1H), 7.89 (d, J = 6.0Hz, 1H), 7.53 (d, J = 9.0Hz, 2H), 6.84 (d, J = 9.1Hz, 2H), 6.21 (d, J = 6.1Hz, 1H),3.92(s,2H),3.23(dt,J=13.7,9.0Hz,2H),3.10–3.02(m,4H),2.66(ddd,J=14.4,11.0,8.1Hz,1H),2.57– 2.52(m,4H),2.28(s,3H),2.13(d,J=10.6Hz,2H),1.91–1.71(m,2H),1.46(td,J=10.5,4.0Hz,2H),1.31(s,3H)

[0124] Example 4: General method for the synthesis of compound 565 (TDM-181165)

[0125] Step 1: Compound 565

[0126] N-(4-methyl-1-(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)piperidin-4-yl)cyclopropanesulfonamide

[0127] To a solution of compound 565a (30 mg, 0.079 mmol) and N,N-diisopropylethylamine (15.2 mg, 0.118 mmol) in dichloromethane (2 mL) was added 4-dimethylaminopyridine (9.6 mg, 0.079 mmol) and compound 565b (19.2 mg, 0.157 mmol) at room temperature. The mixture was heated to 40 degrees Celsius and stirred for 3 hours. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0-75 / 25) and preparative HPLC (formic acid) to give a white solid product (compound 565, 13.3 mg, 34.7% yield). LCMS [M+1] + =486.3.

[0128] 1 H NMR (400MHz, DMSO) δ8.74 (s, 1H), 8.14 (s, 2H), 7.89 (d, J = 6.1 Hz, 1H), 7.53 (d, J = 9. 0Hz,2H),6.86(d,J=4.1Hz,2H),6.84(s,1H),6.21(d,J=6.1Hz,1H),3.79(s,2H),3. 51–3.44(m,2H),3.07(s,4H),2.65–2.60(m,1H),2.57(s,4H),2.30(s,3H),1.93(d, J=14.1Hz,2H),1.57–1.46(m,2H),1.40(s,3H),0.97(d,J=3.4Hz,2H),0.95(s,2H).

[0129] Example 5: General method for the synthesis of compound 572 (TDM-181172)

[0130] Step 1: Compound 572c

[0131] (1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamate

[0132] Potassium carbonate (1.5 g, 10.9 mmol) was added to a solution of compound 572a (1 g, 5.45 mmol) and compound 572b (1.17 g, 5.45 mmol) in acetonitrile (50 mL). The reaction solution was stirred at 50° C. for 1 hour. The LCMS [M+1] +=361.2 The product was detected. The mixture was added to water (50 mL) and extracted with ethyl acetate (2*50 mL). The organic phases were combined, washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (40 g column, 35 minutes) using PE / EA = 0-60% as the eluent to obtain the product as a white solid (compound 572c, 1.65 g, 84% yield). LCMS [M+1] + =361.2.

[0133] Step 2: Compound 572d

[0134] 1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-amine

[0135] Trifluoroacetic acid (10 ml, 134.62 mmol) was added to a solution of compound 572c (1.1 g, 3.05 mmol) in dichloromethane (100 mL) at 0°C, and the mixture was stirred at room temperature for 1 hour. LCMS [M+1] + =261.1 detected product. The mixture was added with water (60 mL), extracted with ethyl acetate (2*60 mL), and the organic layers were combined. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (40 g column, 35 minutes) using DCM / (DCM:MeOH=10:1)=0-50% as eluent to give a yellow solid (compound 572d, 870 mg, crude) as the product. LCMS [M+1] + =261.1.

[0136] Step 3: Compound 572f

[0137] (S)-N-(1-(2,5-dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0138] To a 50 mL flask was added a solution of compound 572d (350 mg, 1.35 mmol), compound 572e (246 mg, 2.02 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (767 mg, 2.05 mmol) in DMF (25 ml), and N,N-diisopropylethylamine (261 mg, 2.01 mmol) was added, and the mixture was stirred at room temperature for 1 hour. By LCMS [M+1] +The product was detected at 365.1. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (40 g column, 35 minutes) using PE / EA = 0-75% to afford the product as a yellow oil (compound 572f, 670 mg, crude). LCMS [M+1] = 365.1.

[0139] Step 4: Compound 572

[0140] (S)-N-(1-(2,5-dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0141] Compound 572f (74.5 mg, 0.2 mmol), compound 572e (45 mg, 0.2 mol), p-toluenesulfonic acid (77 mg, 0.407 mmol) were dissolved in n-butanol (4 mL) in a 50 mL beaker, and the mixture was stirred at 110° C. for 4 hours. LCMS [M+1] + =550.3 product detection. The mixture was added to water (20 mL) and extracted with ethyl acetate (2*20 mL), and the organic layers were combined. The organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by HPLC to give a white solid product (Compound 572, 5.3 mg, 13% yield). LCMS [M+1] + =550.3.

[0142] 1 H NMR(401MHz,DMSO)δ8.20(s,1H),7.98(s,1H),7.95(s,1H),7.72–7.67(m,2H),6.61(d ,J=2.5Hz,1H),6.45(dd,J=8.9,2.5Hz,1H),3.84(d,J=14.4Hz,2H),3.80(s,3H),3.26 –3.20(m,2H),3.14–3.08(m,4H),2.71–2.61(m,1H),2.47–2.42(m,4H),2.23(s,3H),2 .15(d,J=13.0Hz,2H),1.88–1.72(m,2H),1.56(dd,J=13.8,10.4Hz,2H),1.32(s,3H).

[0143] N-(1-(5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide (TDM-181173) was synthesized as a white solid (52.2 mg, 37.17% yield) using a method similar to that of Example 5. (S)-N-(1-(5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide (TDM-181174) was synthesized as a white solid (5 mg, 60% yield).

[0144] Example 6: General Method for Compound 575 (TDM-181175)

[0145] Step 1: Compound 575c

[0146] (1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamate

[0147] Potassium carbonate (1.5 g, 10.9 mmol) was added to a solution of compound 575a (1 g, 5.45 mmol) and compound 575b (1.17 g, 5.45 mmol) in acetonitrile (50 mL). The reaction solution was stirred at 50° C. overnight. The LCMS [M+1] + =361.2 The product was detected. The mixture was added to water (50 mL) and extracted with ethyl acetate (2*50 mL). The organic phases were combined, washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (40 g column, 35 minutes) using PE / EA = 0-60% as the eluent to obtain the product as a white solid (Compound 575c, 1.65 g, 84% yield). LCMS [M+1] + =361.2.

[0148] Step 2: Compound 575d

[0149] 1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-amine

[0150] A hydrochloric acid-dioxane solution (10 ml, 134.62 mmol) was added to a dioxane (100 mL) solution of compound 575c (1.1 g, 3.05 mmol) at 0°C, and the mixture was stirred at room temperature for 1 hour. LCMS [M+1] +=261.1 detected product. The mixture was added with water (60 mL), extracted with ethyl acetate (2*60 mL), and the organic layers were combined. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (40 g column, 35 minutes) using DCM / (DCM:MeOH=10:1)=0-50% as the eluent to give a yellow solid (compound 575d, 870 mg, crude) as the product. LCMS [M+1] + =261.1.

[0151] Step 3: Compound 575f

[0152] N-(1-(2,5-dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0153] To a solution of compound 575d (430.9 g, 1.66 mmol) in anhydrous dichloromethane (20 mL) was added N,N-diisopropylethylamine (321.21 mL, 2.49 mmol), followed by 4-dimethylaminopyridine (202.43 mg, 1.66 mmol) and compound 575e (349.40 mg, 2.49 mmol) at 0°C. The reaction mixture was heated to 40°C and stirred for 3 hours. LCMS [M+1] + =365, indicating the reaction was complete. Post-treatment: The reaction solution was poured into water, and the aqueous phase was extracted with dichloromethane (2*40 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to dryness. The crude product was purified by column chromatography (eluent: EA / PE = 0-30%) to give the target compound (compound 575f, 554.9 mg, 82% yield) as a yellow liquid. LCMS [M+1] + =365.

[0154] Step 4: Compound 575

[0155] N-(1-(5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0156] Compound 575h (109.05 mg, 0.49 mmol), palladium acetate (18.41 mg, 0.082 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (94.89 mg, 0.164 mmol), and cesium carbonate (267.16 mg, 0.82 mmol) were added to compound 575f (150 mg, 0.41 mmol). The reaction solution was purged with argon several times, and anhydrous 1,4-dioxane (10 mL) was added. The reaction solution was purged with argon three times. The temperature was raised to 100°C and stirred for two hours. LCMS [M+1]+ =550.2, indicating the reaction was complete. Post-treatment: The reaction solution was concentrated and dried to obtain a crude product, which was then purified by column chromatography using an eluent of (dichloromethane / methanol = 10:1) / DCM = 0-95%. The crude product was then purified by preparative purification to afford the title compound (Compound 575, 75.7 mg, 33.56% yield) as a white solid. LCMS [M+1] + =550.2.

[0157] 1 H NMR (400MHz, MeOD) δ8.44(s,1H),8.01(d,J=8.8Hz,1H),7.91(d,J=1.7Hz,1H),6.68(d,J=2.3 Hz,1H),6.56(dd,J=8.8,2.3Hz,1H),3.97(d,J=13.1Hz,2H),3.89(s,3H),3.58(t,J=11.8Hz, 2H),3.28(d,J=5.3Hz,4H),3.03(d,J=4.7Hz,4H),2.66(d,J=2.7Hz,3H),2.63–2.54(m,1H),2 .09(d,J=13.9Hz,2H),1.74–1.65(m,2H),1.50(s,3H),1.13–1.07(m,2H),1.06–0.98(m,2H).

[0158] Example 7: General method for the synthesis of compound 578 (TDM-181178)

[0159] Step 1: Compound 578c

[0160] (1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamate

[0161] Potassium carbonate (1.5 g, 10.9 mmol) was added to a solution of compound 578a (1 g, 5.45 mmol) and compound 578b (1.17 g, 5.45 mmol) in acetonitrile (50 mL). The reaction solution was stirred at 50° C. overnight. The LCMS [M+1] + =361.2 The product was detected. The mixture was added to water (50 mL) and extracted with ethyl acetate (2*50 mL). The organic phases were combined, washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (40 g column, 35 minutes) using PE / EA = 0-60% as the eluent to obtain the product as a white solid (compound 578c, 1.65 g, 84% yield). LCMS [M+1] + =361.2.

[0162] Step 2: Compound 578e

[0163] Benzyl-4-(4-((4-(4-(tert-butoxycarbonyl)amino)-4-methylpiperidin-1-yl)-5-chloropyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0164] In a 50 mL flask, compound 578c (251 mg, 0.7 mmol), compound 578d (210 mg, 0.7 mmol), palladium acetate (31.3 mg, 0.14 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (161.26 mg, 0.279 mmol) and cesium carbonate (454 mg, 1.4 mmol) were dissolved in ultra-dry 1,4-dioxane (25 mL), and the mixture was stirred at 100 degrees Celsius under argon for 2 hours. The reaction was monitored by TLC and LCMS. By LCMS [M+1] + =625.3 confirmed product. The mixture was added with water (50 mL), extracted with ethyl acetate (2*50 mL), and the organic layers were combined. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (12 g column, 25 minutes), eluting with DCM / (DCM:MeOH=10:1)=0-50% to obtain a white solid (compound 578e, 240 mg, 50% yield) product LCMS [M+1] + =625.3.

[0165] Step 3: Compound 578f

[0166] Benzyl 4-(4-((4-(4-amino-4-methylpiperidin-1-yl)-5-chloropyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0167] Compound 578e (100 mg, 0.16 mmol) was dissolved in dichloromethane (10 mL) at 0°C, trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC and LCMS. By LCMS [M+1] + =525.3 product was detected. The reaction mixture was concentrated under reduced pressure, poured into water (30 mL), and adjusted to pH>8 with potassium carbonate (50%). Water (30 mL) and ethyl acetate (2*30 mL) were added to extract the mixture, and the organic layers were combined. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow solid as the product (compound 578f, 170 mg, crude). LCMS [M+1] + =525.3.

[0168] Step 4: Compound 578h

[0169] Benzyl (S)-4-(4-((5-chloro-4-(2-(2,2-difluorocyclopropane-1-carboxamido)-4-methylpiperidin-1-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0170] Take a 50mL flask, compound 578f (140mg, 0.27mmol), compound 578g (48.8mg, 0.4mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (152mg, 0.4mmol), dissolve in N,N-dimethylformamide (15mL), add N,N-diisopropylethylamine (51.6mg, 0.4mmol), and stir the mixture at room temperature for 1 hour. The reaction was detected by TLC and LCMS. The product was confirmed by LCMS [M+1] + = 629.2. The mixture was added with water (30mL), extracted with ethyl acetate (2*30mL), and the organic layers were combined. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The organic layer was concentrated and purified by flash chromatography (12 g column, 25 minutes) using DCM / (DCM:MeOH=10:1) = 0-100% as eluent to afford the target product as a yellow oil (compound 578h, 133 mg, 74% yield). LCMS [M+1] + =629.2.

[0171] Step 5: Compound 578

[0172] (S)-2,2-Difluoro-N-(4-methyl-1-(5-methyl-2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)piperidin-4-ylcyclopropane-1-carboxamide

[0173] Take a 25 mL flask, add compound 578h (40 mg, 0.064 mmol) at 0°C, cool for a while, then add a solution of hydrobromic acid in acetic acid (2 ml) dropwise, and stir the mixture at 0°C for 0.5 h. The reaction is monitored by TLC and LCMS. By LCMS [M+1] + =495.2 The product was detected. The mixture was concentrated under reduced pressure. The crude product was purified by HPLC to give a white solid product (Compound 578, 12 mg, 37.5% yield). LCMS [M+1] + =495.2.

[0174] 1H NMR(401MHz,DMSO)δ9.24(s,1H),8.29(s,1H),8.01(s,2H),7.78(s,1H),7.5 2(s,1H),4.26(s,1H),3.85(d,J=8.6Hz,2H),3.31(s,2H),3.20(d,J=12.0Hz ,2H),2.80(t,J=11.5Hz,2H),2.72–2.63(m,1H),2.19(d,J=11.3Hz,2H),2.0 3(d,J=11.8Hz,2H),1.97–1.64(m,5H),1.60(t,J=11.9Hz,2H),1.33(s,3H).

[0175] Example 8: General method for the synthesis of compound 582 (TDM-181182)

[0176] Step 1: Compound 582c

[0177] 2,5-Dichloro-4-vinylpyrimidine

[0178] To a solution of compound 582a (2.01 g, 10.96 mmol) in dioxane (100 mL) were added compound 582b (1.69 g, 10.96 mmol), 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (800 mg, 1.1 mmol), cesium carbonate (7.14 g, 21.92 mmol), and water (10 mL). The reaction mixture was purged with argon several times and heated to 70°C for 1 hour, whereupon the reaction was determined to be complete. Workup: The reaction mixture was poured into water, and the aqueous phase was extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled to dryness. The crude product was purified by column chromatography (eluent: EA / PE = 0-5%) to afford the title compound (compound 582c, 960 mg, 47% yield) as a yellow oil. LCMS [M+1] + =175,177.

[0179] Step 2: Compound 582d

[0180] 2,5-Dichloropyrimidine-4-carbaldehyde

[0181] Compound 582c (700 mg, 60% purity) was dissolved in dichloromethane (15 mL), and the solution was cooled to -78°C and purged with ozone for 30 minutes. Dimethyl sulfide was then added and stirred for 3 minutes. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (ethyl acetate / petroleum ether = 0 to 30 / 70) to give a white solid product (compound 582d, 323 mg, crude product). LCMS [M+1] +=177&179.

[0182] Step 3: Compound 582f

[0183] tert-Butyl (1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0184] Compound 582e (430 mg, 2.008 mmol) and acetic acid (2 drops) were added to a solution of compound 582d (323 mg, 1.825 mmol) in dichloromethane (18 mL) at room temperature, and the mixture was stirred for 1 hour. Sodium acetate borohydride (580 mg, 2.738 mmol) was added, and the mixture was stirred for 20 minutes, then concentrated under reduced pressure. The residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0 to 15 / 85) to give a white solid product (compound 582f, 350 mg, 51.1% yield). LCMS [M+1] + =375&377.

[0185] Step 4: Compound 582h

[0186] tert-Butyl (1-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0187] To a mixture of compound 582f (125 mg, 0.342 mmol), compound 582g (72 mg, 0.376 mmol), palladium acetate (15 mg, 0.0684 mmol), xantphos (79 mg 1.1318 mmol) and cesium carbonate (223 mg, 0.684 mmol) was added dioxane (10 mL) at room temperature, and the mixture was heated to 100° C. and stirred for 2 hours. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane=0-80 / 20) to give a white solid product (compound 582h, 125 mg, 69.1% yield). LCMS [M+1] + =530.

[0188] Step 5: Compound 582i

[0189] 4-((4-amino-4-methylpiperidin-1-yl)methyl)-5-chloro-N-(4-(4-methylpiperazin-1-yl)phenyl)pyrimidin-2-amine

[0190] To a solution of compound 582h (125 mg, 0.236 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (1.5 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. The mixture was added with water and neutralized with aqueous potassium carbonate solution, extracted with ethyl acetate, and the organic layers were combined and dried over sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was used directly in the next reaction. LCMS [M+1] + =430.

[0191] Step 6: Compound 582

[0192] (S)-N-(1-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0193] To a solution of compound 582i (50 mg, 0.116 mmol) in N,N-dimethylformamide (5 mL) was added compound 582j (21 mg, 0.174 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (66 mg, 0.174 mmol) and N,N-diisopropylethylamine (22.5 mg, 0.174 mmol) at room temperature, and the mixture was stirred for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0-100 / 0) and preparative HPLC (formic acid) to give a yellow solid product (compound 582, TDM-181182, 8.8 mg, 14.2% yield). LCMS [M+1] + =534.

[0194] 1 H NMR (400MHz, DMSO) δ10.44(s,1H),9.74(s,1H),8.49(s,1H),8.14(s,1H),7.57(d,J=7.9Hz,2H),6.97(d,J=8.8Hz,2H),4.57(s,2 H),3.68(s,4H),3.45(s,2H),2.99(s,6H),2.82(s,3H),2.65(d,J=12.9Hz,2H),2.33(s,2H),1.83(d,J=6.7Hz,4H),1.31(s,3H).

[0195] Example 9: General method for the synthesis of compound 583 (TDM-181183)

[0196] Step 1: Compound 583c

[0197] 2,5-Dichloro-4-vinylpyrimidine

[0198] To a solution of compound 583a (2.01 g, 10.96 mmol) in dioxane (100 mL) were added compound 583b (1.69 g, 10.96 mmol), 1,1'-bis(diphenylphosphinoferrocenepalladium) dichloride (800 mg, 1.1 mmol), cesium carbonate (7.14 g, 21.92 mmol), and water (10 mL). The reaction mixture was purged with argon several times and heated to 70°C for 1 hour, whereupon the reaction was determined to be complete. Workup: The reaction mixture was poured into water, and the aqueous phase was extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled to dryness. The crude product was purified by column chromatography (eluent: EA / PE = 0-5%) to afford the title compound (compound 583c, 960 mg, 47% yield) as a yellow oil. LCMS [M+1] + =175,177.

[0199] Step 2: Compound 583d

[0200] 2,5-Dichloropyrimidine-4-carbaldehyde

[0201] A solution of compound 583c (960 mg, 5.48 mmol) in dichloromethane (130 mL) was added to a 100 mL three-necked flask. The reaction mixture was cooled to -78°C and a sufficient amount of ozone was introduced. The reaction mixture was allowed to react for 15 minutes until completion was detected. Post-treatment: Dimethyl sulfide was added dropwise at -78°C, the temperature was raised to room temperature, and the mixture was stirred for 30 minutes. The mixture was concentrated to dryness, and the crude product was purified by column chromatography (eluent: EA / PE = 0-30%) to afford the title compound (compound 583d, 90 mg, 13.3% yield) as a yellow oil. LCMS [M+1] + =177,179.

[0202] Step 3: Compound 583f

[0203] tert-Butyl (1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0204] To a solution of compound 583d (90 mg, 0.51 mmol) in dichloromethane (10 mL) were added compound 583e (109 mg, 0.51 mmol) and acetic acid (1 drop). The reaction solution was stirred at room temperature for 1.5 hours, followed by the addition of sodium acetate borohydride (162.1 mg, 0.77 mmol) and continued stirring for 30 minutes. The reaction was detected to be complete. Post-treatment: The reaction solution was poured into water and extracted three times with ethyl acetate (3*50 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and dried. The crude product was purified by column chromatography (eluent: EA / PE = 0-30%) to give the title compound (compound 583f, 76 mg, 53% yield) as a yellow solid. LCMS [M+1] + =375,377.

[0205] Step 4: Compound 583g

[0206] 1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-amine

[0207] To a solution of compound 583f (100 mg, 0.27 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for 1 hour, and the reaction was detected to be complete. Post-treatment: The reaction mixture was poured into water, and ethyl acetate (1*50 mL) was added to extract and remove impurities. The aqueous phase was adjusted to pH = 11 with potassium carbonate, and extracted three times with ethyl acetate (3*80 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled to dryness to obtain the target compound (compound 583g, 86.5 mg, 93.3% yield) as a yellow solid. LCMS [M+1] + =275,277.

[0208] Step 5: Compound 583i

[0209] N-(1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0210] To a solution of compound 583g (86.5 mg, 0.31 mmol) in dichloromethane (8 mL) were added 4-dimethylaminopyridine (38.4 mg, 0.31 mmol), N,N-diisopropylethylamine (81.3 mg, 0.63 mmol), and compound 583h (88.4 mg, 0.63 mmol). The reaction mixture was heated to 40°C and stirred for 3 hours. The reaction was determined to be complete. Post-treatment: The reaction mixture was poured into water and extracted with ethyl acetate (3 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled to dryness. The crude product was purified by column chromatography (eluent: (dichloromethane:methanol = 10:1) / dichloromethane = 0-20%) to afford the title compound (compound 583i, 49.6 mg, 34% yield) as a yellow solid. LCMS [M+1] + =379,381.

[0211] Step 6: Compound 583

[0212] N-(1-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0213] To a 100 mL three-necked flask were added compound 583i (49.6 mg, 0.13 mmol), compound 583j (22.5 mg, 0.12 mmol), palladium acetate (5.8 mg, 0.03 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (30 mg, 0.05 mmol), cesium carbonate (84.7 mg, 0.26 mmol), and 1,4-dioxane (5 mL). The reaction solution was purged with argon several times, heated to 100°C, and stirred for 2 hours. The reaction was determined to be complete. Post-treatment: The reaction solution was directly drained, and the crude product was purified by column chromatography (eluent: (dichloromethane:methanol = 10:1 / dichloromethane = 0-100%) to obtain the target compound (compound 583, 8.6 mg, 12.4% yield) as a yellow solid. LCMS [M+1] + =534.2.

[0214] 1H NMR (400MHz, DMSO-d6) δ9.57(s,1H),8.38(s,1H),8.16(s,2H),7.58(d,J=8.9Hz ,2H),6.94–6.83(m,2H),6.68(s,1H),3.53(s,2H),3.06(t,J=5.0Hz,4H),2.65–2 .58(m,2H),2.58–2.52(m,4H),2.47(d,J=4.7Hz,4H),2.24(s,3H),1.91(d,J=13. 2Hz, 2H), 1.53 (ddd, J=12.9, 8.5, 3.6Hz, 2H), 1.33 (s, 3H), 0.92 (d, J=6.4Hz, 4H).

[0215] N-(1-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide (TDM-181184) was synthesized as a yellow solid (8.8 mg, 12.4% yield) using a similar method to Example 9.

[0216] Example 10: General method for the synthesis of compound 585 (TDM-181185)

[0217] Step 1: Compound 585c

[0218] tert-Butyl (1-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0219] To a mixture of compound 585a (116 mg, 0.294 mmol), compound 585b (62 mg, 0.279 mmol), palladium acetate (13 mg, 0.0588 mmol), xantphos (68 mg, 0.1176 mmol) and cesium carbonate (191 mg, 0.588 mmol) was added dioxane (10 mL) at room temperature, and the mixture was heated to 100° C. and stirred for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0 to 80 / 20) to give a white solid product (compound 585c, 145 mg, 87.9% yield). LCMS [M+1] + =560.

[0220] Step 2: Compound 585d

[0221] 4-((4-amino-4-methylpiperidin-1-yl)methyl)-5-chloro-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyrimidin-2-amine

[0222] To a solution of compound 585c (145 mg, 0.315 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (1.5 mL) at room temperature, and the mixture was stirred at 20°C for 1 hour. The mixture was added with water and neutralized with aqueous potassium carbonate solution, extracted with ethyl acetate, and the organic layers were combined and dried over sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was used directly in the next reaction. LCMS [M+1] + =460.

[0223] Step 3: Compound 585

[0224] (S)-N-(1-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0225] To a solution of compound 585d (55 mg, 0.120 mmol) in N,N-dimethylformamide (5 mL) was added compound 585e (21.9 mg, 0.179 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (68 mg, 0.179 mmol) and N,N-diisopropylethylamine (23 mg, 0.179 mmol) at room temperature, and the mixture was stirred for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0-100 / 0) and preparative HPLC (formic acid) to give the product as a yellow solid (compound 585, TDM-181185, 10 mg, 14.8% yield). LCMS [M+1] + =564.3.

[0226] 1H NMR (400MHz, DMSO) δ8.31(s,1H),8.24(s,1H),8.15(s,2H),7.75(s,1H),7.59(d,J=8.7Hz,1H ),6.62(d,J=2.4Hz,1H),6.47(dd,J=8.8,2.4Hz,1H),3.77(s,3H),3.49(s,2H),3.18–3.11(m, 4H),2.67(td,J=6.5,4.4Hz,1H),2.62(dd,J=8.8,5.8Hz,2H),2.57–2.54(m,4H),2.42–2.34(m ,2H),2.30(s,3H),2.05(s,2H),1.88–1.70(m,2H),1.47(dd,J=11.2,6.6Hz,2H),1.25(s,3H).

[0227] Example 11: General method for the synthesis of compound 586 (TDM-181186)

[0228] Step 1: Compound 586b

[0229] 1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-amine

[0230] To a solution of compound 586a (116 mg, 0.309 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (2 mL) at room temperature, and the mixture was stirred for 1 hour. The mixture was added with water and neutralized with aqueous sodium hydroxide solution, extracted with ethyl acetate, the organic layers were combined, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (10% methanol (NH3) in dichloromethane / dichloromethane = 0-100 / 0) to give a white solid product (compound 586b, 68 mg, 80% yield). LCMS [M+1] + =275&277.

[0231] Step 2: Compound 586d

[0232] (S)-N-(1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0233] To a solution of compound 586b (48 mg, 0.1475 mmol) in N,N-dimethylformamide (5 mL) were added compound 586c (32 mg, 0.262 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (100 mg, 0.262 mmol) and N,N-diisopropylethylamine (34 mg, 0.262 mmol) at room temperature, and the mixture was stirred for 1 hour. The mixture was added with water and extracted with ethyl acetate. The organic layers were combined, washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0-15 / 85) to give a white solid product (compound 586d, 56.8 mg, 85.6% yield). LCMS [M+1] + =379&381.

[0234] Step 3: Compound 586f

[0235] Benzyl (S)-4-(4-((5-chloro-4-((4-(2,2-difluorocyclopropane-1-carboxamido)-4-methylpiperidin-1-yl)methyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0236] To a solution of compound 586d (47 mg, 0.124 mmol) in dioxane (5 mL) were added compound 586e (41 mg, 0.136 mmol) and p-toluenesulfonic acid monohydrate (47 mg, 0.248 mmol) at room temperature, and the mixture was heated to 100° C. and stirred for 24 hours. The mixture was added with water and extracted with ethyl acetate. The organic layers were combined, washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0 to 50 / 50) to give a yellow solid product (compound 586f, 21 mg, 26.3% yield). LCMS [M+1] + =643.

[0237] Step 4: Compound 586

[0238] (S)-N-(1-((5-chloro-2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0239] Compound 586f (21 mg, 0.033 mmol) was dissolved in HBr-AcOH (1.5 mL) at 0°C, and the mixture was stirred at 0°C for 30 minutes. The mixture was concentrated under reduced pressure at 45°C, and the residue was purified by preparative HPLC (formic acid) to give a gray solid product (compound 586, TDM-181186, 6.7 mg, 40.4% yield). LCMS [M+1] + =509.

[0240] 1 H NMR(400MHz,DMSO)δ9.74(s,1H),8.39(s,1H),8.23(s,2H),7.96(s,1H),7.79(s,1H) ,7.59(s,1H),4.39(d,J=10.4Hz,1H),4.10–3.99(m,1H),3.54(s,2H),3.34(d,J=12. 3Hz,2H),2.98(t,J=11.0Hz,2H),2.73–2.66(m,1H),2.65–2.55(m,2H),2.39(s,2H), 2.08(dd,J=27.2,12.6Hz,6H),1.88–1.70(m,2H),1.48(d,J=8.9Hz,2H),1.26(s,3H).

[0241] Test example: Enzyme activity inhibition assay of NUAK1 / NUAK2 kinase inhibitors

[0242] The inhibitory effect of the pyrimidine amine small molecules involved in this application on the NUAK1 and NUAK2 kinases was determined using a kinase activity assay based on P81 filter paper combined with hotspot technology (Nat Biotechnol. 2011, 29: 1039), which is briefly described as follows:

[0243] The assay buffer system contained 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.01% Brij35 (L23 polyoxyethylene lauryl ether), 0.02 mg / ml BSA (bovine serum albumin), 0.1 mM Na3VO4, 2 mM DTT, and 1% DMSO.

[0244] Test steps:

[0245] 1. Prepare a 20 μM substrate solution using freshly prepared assay buffer. The substrate is a CHKtide polypeptide fragment (Sanchez Y. Science. 1997, 277: 1497-1501).

[0246] 2. Add kinase and mix gently (the final concentration of NUAK1 is 10 nM; the final concentration of NUAK2 is 50 nM);

[0247] 3. Add the test compound dissolved in 100% DMSO to the above kinase reaction mixture using acoustic pipetting technology (Echo550; nanoliter) and incubate at room temperature for 5 minutes;

[0248] 4. Join 33 P-labeled ATP (unlabeled ATP / labeled ATP ratio of 25:1 or 2.5:1);

[0249] 5. Incubate at room temperature for 2 hours;

[0250] 6. Detect kinase activity using P81 filter paper combined with hotspot technology.

[0251] The IC of the test compound against NUAK1 / NUAK2 was calculated according to the above assay. 50 , see Table 1 for specific results.

[0252] IC 50 Calculations were made using the formula derived from a sigmoidal dose-response curve (variable slope):

[0253] Y = Bottom + (Top - Bottom) / (1 + 10^((LogEC50 - X) * slope, where X is the Log value of the compound concentration and Y is the response (inhibition rate of kinase activity). Y rises from the bottom to the top along the S-shaped curve as the concentration increases.

[0254] Table 1 IC values ​​of the test compounds of this application for NUAK1 / NUAK2 50 (nM)

[0255] From the results in Table 1, it can be seen that the pyrimidine amine compound of the present application has excellent inhibitory activity against both NUAK1 and NUAK2, and is a dual-target small molecule kinase inhibitor. The IC value of the compound for NUAK1 is 50 The IC value for NUAK2 can be lower than 1nM. 50 It can reach several nM or tens of nM. Therefore, the above experiments have proved that the pyrimidine amine compounds of the present application can be used as NUAK1 / NUAK2 inhibitors.

[0256] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0257] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0258] In addition, the various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. A pyrimidine amine compound, characterized in that The pyrimidine amine compound is a compound represented by formula I, or a stereoisomer, tautomer, isotope derivative, hydrate, solvate, prodrug, and pharmaceutically acceptable salt thereof; Wherein, X is a direct bond, -O-, -S-, -NR7- or -CR8R9-, and Y is -CO- or -S(O)2-; A is an optionally substituted 6-10 membered aryl or 5-10 membered heteroaryl, B is an optionally substituted 3-10 membered cycloalkyl or 3-10 membered heterocycloalkyl, and the substituents on A or B are selected from one or more of halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino; R1, R2, R6, R7, R8, and R9 are each independently H, or an optionally substituted C1-8 alkyl group; R3 and R4 are each independently halogen, amino, hydroxyl, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, or optionally substituted C1-8 alkylamino; R5 is an optionally substituted 3-10 membered cycloalkyl, an optionally substituted 3-10 membered heterocycloalkyl, wherein the substituent on the cycloalkyl or heterocycloalkyl is selected from one or more of halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino; The substituent on the C1-8 alkyl group is selected from one or more of halogen, amino, hydroxyl, nitro, cyano, and mercapto; n is 0, 1 or 2, and m is 0, 1, 2, 3, 4 or 5.

2. The compound according to claim 1, characterized in that R5 is an optionally substituted 3-10 membered cycloalkyl group.

3. The compound according to claim 2, characterized in that R5 is an optionally substituted 3-6 membered cycloalkyl group.

4. The compound according to claim 3, characterized in that R5 is a 3-6 membered cycloalkyl group substituted with halogen.

5. The compound according to claim 4, characterized in that R5 is a fluorine-substituted 3-6-membered cycloalkyl group.

6. The compound according to claim 3, characterized in that R5 is 7. The compound according to claim 6, characterized in that R5 is 8. The compound according to any one of claims 1 to 7, characterized in that R6 is methyl.

9. The compound according to any one of claims 1 to 8, characterized in that R2 is H.

10. The compound according to any one of claims 1 to 9, characterized in that B is an optionally substituted 3-6-membered heterocycloalkyl group.

11. The compound according to claim 10, characterized in that B is an optionally substituted 5-6-membered heterocycloalkyl group.

12. The compound according to claim 11, characterized in that B is optionally substituted piperidinyl or optionally substituted piperazinyl.

13. The compound according to claim 12, characterized in that The compound represented by formula I is formula I-1 or formula I-2, R 10 is selected from halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino; R 11 is H or optionally substituted C1-8 alkyl; The substituent on the C1-8 alkyl group is selected from one or more of halogen, amino, hydroxyl, nitro, cyano, and mercapto; q is 0, 1, 2, 3 or 4.

14. The compound according to claim 13, characterized in that R 11 It is H or methyl.

15. The compound according to any one of claims 1 to 14, characterized in that A is optionally substituted phenyl or optionally substituted 5-membered heteroaryl.

16. The compound according to claim 15, characterized in that A is one of the following groups which are optionally substituted:

17. The compound according to any one of claims 1 to 16, characterized in that R3 is chlorine, preferably substituted at the para position of the amino group.

18. The compound according to any one of claims 1 to 17, characterized in that The substituent on the A ring is a methoxy group, preferably substituted at the ortho position of the amino group.

19. The compound according to any one of claims 1 to 18, characterized in that The compound represented by the formula I is the following compound:

20. The compound according to any one of claims 1 to 19, wherein the pharmaceutically acceptable salt is a formate salt.

21. The compound according to any one of claims 1 to 20, wherein the isotopic derivative is a deuterated compound.

22. A method for preparing a compound according to any one of claims 1 to 21, comprising the following steps: The compound of formula I is prepared from the compound of formula II and the compound of formula III; or The compound of formula V is prepared from the compound of formula IV and formula II, and the compound of formula I is prepared from the compound of formula V; Optionally, the preparation process includes necessary protection and deprotection steps; in, Z is a leaving group, preferably Z is halogen, more preferably Z is Cl, and the definitions of other groups are the same as those in claims 1-21.

23. A pharmaceutical composition, characterized in that The compound according to any one of claims 1 to 21 is used as an active ingredient.

24. The pharmaceutical composition according to claim 23, characterized in that Contains a pharmaceutically acceptable carrier or excipient.

25. Use of the compound according to any one of claims 1 to 21 for preparing a NUAK1 or NUAK2 inhibitor.

26. Use of the compound according to any one of claims 1 to 21 for preparing a medicament, characterized in that The compound is used for preventing or treating the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases.

27. The use according to claim 26, characterized in that The disease is Parkinson's disease, Alzheimer's disease, diabetes, hyperlipidemia, obesity, liver cancer, leukemia, lymphoma, tumor metastasis, cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or is used alone to reduce trauma and postoperative scars.

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