A pyrimidineamine NUAK inhibitor, its preparation method and uses

Abstract

The present application provides a pyrimidineamine compound, characterized in that the pyrimidineamine compound is a compound represented by Formula I, or its stereoisomer, tautomer, isotope derivative, hydrate, solvate, prodrug, and pharmaceutically acceptable salt. The pyrimidineamine compound described in the present application has 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 fibrosis diseases, and skin fibrosis diseases. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of small molecule compounds, and specifically relates to a pyrimidineamine-based NUAK inhibitor, its preparation method and uses. The compound can be used for preventing or treating the following diseases by inhibiting NUAK1 / NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, skin fibrosis diseases, or can be used alone to reduce trauma and postoperative scars. Background Art

[0002] Protein kinases are a group of important functional proteins involved in regulating cell metabolism, polarity, growth, division and differentiation. The human genome encodes more than 500 protein kinases, and these kinases can transfer the phosphate group on ATP (adenosine triphosphate) to specific serine, threonine or tyrosine residues of substrate proteins to phosphorylate them; most protein kinases are serine / threonine kinases, and there are less than 100 tyrosine kinases. Adenosine monophosphate-activated protein kinase (AMPK) belongs to serine / threonine kinases (STK) and is an important regulator of cellular energy homeostasis in mammals. It regulates glucose and lipid metabolism, cell proliferation and cell polarity by sensing the changes in the ratio of intracellular AMP (Adenosine monophosphate) / ATP or ADP (adenosine diphosphate) / ATP under metabolic stress (such as hypoxia, heat shock), and can be called a metabolic sensor protein. AMPK is very conserved in evolution and is a heterotrimeric protein composed of an α subunit containing a kinase domain (KD), a β subunit and a γ subunit that regulate kinase activity. The γ subunit contains four CBS functional domains (Cystathionine-β-synthase (CBS) domains) responsible for detecting the changes in the ratio of intracellular AMP / ATP and ADP / ATP (Hardie DG, Trends in Cell Biology. 2016, 26:190).

[0003] Dysfunction of AMPK can lead to various diseases such as obesity, diabetes, inflammatory diseases, and tumors. Therefore, the regulation of AMPK function by the body is very crucial. First of all, the activation of AMPK requires the phosphorylation of threonine (T172) at position 172 of its α subunit by upstream kinases. Currently, there are three known upstream kinases of AMPK: liver kinase B1 (LKB1), calcium / calmodulin-dependent protein kinase 2 (Ca 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 the T172 site, there are three known protein phosphatases that can inhibit the activity of AMPK by removing the phosphate group on T172, including protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), and magnesium / manganese-dependent protein phosphatase 1E (Mg 2+ - / Mn 2+ -dependent protein phosphatase 1E, PPM1E). When the cell is in a low-energy state (high AMP / ATP or ADP / ATP ratio), the KD of the α subunit and the γ subunit of AMPK are tightly cross-linked, and at the same time, the β subunit is myristoylated, ensuring that the protein phosphatase cannot access the T172 site and remains in the activated state. When the cell is in a high-energy state, the KD and γ subunit loosen, and T172 is exposed to the phosphatase, resulting in the inactivation of AMPK (Steinberg GR, Nature Reviews Drug Discovery. 2019, 18:527).

[0004] In addition to the fact that removing AMPK upstream kinases and protein phosphatases can activate and inactivate AMPK respectively, there is also a class of AMPK-related kinases (ARKs) that participate in regulating the function of AMPK. So far, a total of 12 ARKs (BRSK1, BRSK2, NUAK1, NUAK2, QIK, QSK, SIK, MARK1, MARK2, MARK3, MARK4, and MELK) have been discovered. They all belong to serine / threonine protein kinases. The kinase domains of ARKs have a high homology with the α subunit of AMPK. Except for MELK, they can all be activated by LKB1, and the phosphorylation sites for kinase activation are also equivalent to T172 of AMPK. Functionally, they all participate in the regulation of cell metabolism, proliferation, and polarity. However, since ARKs do not have regulatory subunits like AMPK, they cannot be directly regulated by the intracellular AMP / ATP ratio (Bright NJ, Acta Physiologica. 2009, 196:15).

[0005] ARKs are further divided into several ARK subfamilies according to different protein structures and functional characteristics. Among them, 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 are approximately 55% homologous. According to the amino acid sequences, the predicted molecular weights of NUAK1 and NUAK2 are 76 and 69 kDa respectively; their protein structures are very similar. The amino terminus is the kinase domain, and the carboxyl terminus is the ubiquitin-associated domain. It is not yet clear whether NUAK, like AMPKs, is also an heterotrimeric structure. NUAK1 and NUAK2 are expressed in most tissues. Among them, the expression of NUAK1 in organs and tissues such as the brain, skin, muscle, upper digestive tract, and endocrine is significantly higher than other parts. The organs and tissues with the highest expression of NUAK2 are the digestive tract, female reproductive system, skin, bone, brain, and endocrine, etc. Among them, the expression of NUAK2 shows more tissue specificity. It is worth mentioning that NUAK1 and other ARKs and AMPK proteins are mainly distributed in the cytoplasm, while NUAK2 is mainly distributed in the nucleus, and there are indications that NUAK2 participates in the gene expression related to metabolic stress as a transcriptional regulator (Sun X, J of Molecular Endocrinology. 2013, 51:R15).

[0006] As serine / threonine protein kinases, NUAK1 and NUAK2 can phosphorylate a variety of protein substrates, including proteins involved in cell signaling, metabolism, cell proliferation, apoptosis, autophagy, and cytoskeleton organization. NUAK1 is known to phosphorylate AMPK, LATS1 / 2, the p53 tumor suppressor protein, and myosin phosphatase target subunit 1 (Mypt1), which regulates actin cytoskeleton organization, etc. NUKA2 can phosphorylate the transcription factor Gli3, FoxO1, kinesin light chain 1 (KLC1), and Rho GDP dissociation inhibitor α (Rho GDIα) in the Hedgehog signaling pathway, etc.

[0007] The functional regulation of NUAK1 and NUAK2 involves multiple intracellular signaling systems. In addition to being phosphorylated and activated by LKB1 (T211, equivalent to T172 of AMPK) and activated by calcium / PKC, NUAK1 is the only member of the AMPK-related protein family that can be activated by Akt. An increase in NUAK1 activity is also seen in the activation of some growth factor signaling pathways such as insulin-like growth factor 1 (IGF1), and the activity of NUAK1 is enhanced when skeletal muscle cells are in a contracted state. Both NUAK1 and NUAK2 can be activated by LKB1 (T211 / NUAK1, T208 / NUAK2, equivalent to T172 of AMPK), but NUAK2 can be autophosphorylated and activated. NUAK2 can interact with ubiquitin-specific protease 9 on the X chromosome (USP9X), which is a deubiquitinating enzyme that can maintain the activity of NUAK2. In different cells, stimuli such as low osmotic pressure, DNA damage, oxidation, and nutrient deficiency can lead to the activation of NUAK2, and the activation of NUAK2 is also seen in the contraction of skeletal muscle cells (Brooks D, Data in Brief. 2022, 43:108482).

[0008] Multiple studies have shown that NUAK plays an important role in the pathogenesis of diseases such as metabolic diseases, tumors, neurodegenerative diseases, and fibrotic diseases. Homozygous knockout of NUAK1 and NUAK2 genes in mice basically leads to embryonic death. Heterozygous deletion of NUAK1 in mice results in dysplasia of the body and nervous tissues (such as cerebral cortex and peripheral neurons). Heterozygous mutations in the human NUAK1 gene are associated with autism spectrum disorders (ASD), cognitive deficits, attention deficit / hyperactivity disorder (AD / HD), and schizophrenia. Deletion of the human NUAK2 gene leads to the formation of anencephaly, a severe neural tube defect that causes fetal developmental defects, and its mechanism is related to the loss of function of YAP (Bonnard C, J Exp Med. 2020, 217:e20191561). Specific knockout of the NUAK1 gene in skeletal muscle cells can prevent subclinical diabetes induced by a high-fat diet; while the phenotypes of NUAK2 heterozygous deletion mice are similar to a series of clinical manifestations of human type 2 diabetes with obesity, and these phenomena may be related to the imbalance of the autophagy mechanism (Bennison SA, Cellular Signaling. 2022, 100:110472; Blazejewski SM, Scientific Reports. 2011, 11:8156).

[0009] Activation of NUAK1 by Akt and other protein kinases can promote the survival of tumor cells in an energy-deficient environment and protect tumor cells from apoptosis. NUAK2 also inhibits TNFα- and CD95-induced apoptosis through a similar mechanism. In addition, after activation, NUAK1 promotes the invasion and metastasis of tumor cells by upregulating matrix metalloproteinases (MMPs), while NUAK2 participates in the occurrence of tumor metastasis by enhancing the motility of tumor cells (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). A lot of evidence shows that NUAK1 and NUAK2 play a role in tumor formation and metastasis. Compared with NUAK1, NUAK2 has a stronger effect on promoting tumor formation and metastasis. NUAK2 inhibitors should be more likely to become new targeted anti-tumor drugs than NUAK1 inhibitors.

[0010] Recent studies have shown that NUAK plays a key role in the development of tissue fibrosis through its interaction with the transforming growth factor-β (TGF-β) signaling pathway. First, TGF-β can upregulate the gene transcription of NUAK1 and NUAK2 in a variety of epithelial cells such as keratinocytes and human dermal fibroblasts, while the inactivation of the MAPK (ERK1 / 2 and p38) signaling inhibits TGF-β-dependent NUAK2 expression. In addition, NUAK2 inhibits the intracellular degradation of SMAD3 by binding to the structural cross-linking domain and MH2 domain of the intracellular signal mediator SMAD3 protein of TGF-β. A similar mechanism also exists between NUAK2 and TβRI. The gene expression of pro-fibrotic molecules such as fibronectin (FN), plasminogen activator inhibitor 1 (PAI1), and tissue inhibitor of metalloproteinase-1 (TIMP1) induced by TGF-β depends on the presence of NUAK2, indicating that NUAK2 promotes the occurrence of fibrosis. Studies have also shown that NUAK1 promotes renal, pulmonary, and hepatic fibrosis by upregulating the TGF-β and YAP signaling pathways. In animal experiments, inhibiting NUAK1 can reduce scars caused by new wounds and old scar tissues (Gill MK, Nature Communications. 2018.9:3510). Interestingly, it was found that the gene expression of FN encoding was upregulated in NUAK1-knockout keratinocytes, and NUAK1 may affect TGF-β signal transduction through a negative feedback mechanism to inhibit fibrosis (van de Vis RAJ, Cancers. 2021, 13:3377). Generally speaking, inhibiting NUAK may effectively inhibit the fibrosis process of tissues and organs.

[0011] Since kinases have always been ideal small molecule drug targets in terms of structure and function, and NUAK1 and NUAK2 are involved in the occurrence of various diseases, there are currently some NUAK inhibitors in the early stage of research and development, but their effectiveness and safety remain doubtful (Banerjee S, The Biochemical Journal. 2014, 457: 215). Therefore, the development of NUAK selective inhibitors and NUAK1 / 2 dual-target inhibitors is expected to provide new directions for innovative treatments for 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 liver cirrhosis, renal fibrosis, pulmonary fibrosis, and sequelae of myocarditis), and skin fibrosis diseases (such as scleroderma, keloid, hypertrophic scar, or simply used to reduce scars after trauma and surgery). Developing a new generation of NUAK inhibitors has great potential clinical application value. Summary of the Invention

[0012] The object of the present invention is to obtain effective NUAK1 / NUAK2 inhibitors, which 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 liver cirrhosis, renal fibrosis, pulmonary fibrosis, and sequelae of myocarditis), and skin fibrosis diseases (such as scleroderma, keloid, hypertrophic scar, or simply used to reduce scars after trauma and surgery).

[0013] To achieve the above object, on the one hand, the present invention provides a pyrimidineamine compound, which is a compound represented by the following formula I, or its stereoisomer, geometric isomer, tautomer, isotope derivative, hydrate, solvate, prodrug, and pharmaceutically acceptable salt:

[0014]

[0015] Wherein, X is a direct bond, -O-, -S-, -NR7-, or -CR8R9-; Y is -CO- or -S(O)2-;

[0016] 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; 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 alkoxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino;

[0017] R1, R2, R6, R7, R8, and R9 are each independently H or an optionally substituted C1-8 alkyl group;

[0018] R3 and R4 are each independently a halogen atom, an amino group, a hydroxyl group, a nitro group, a cyano group, a mercapto group, an optionally substituted C1-8 alkyl group, an optionally substituted C1-8 alkoxy group, an optionally substituted C1-8 alkylthio group, or an optionally substituted C1-8 alkylamino group;

[0019] R5 is an optionally substituted 3- to 10-membered cycloalkyl group or an optionally substituted 3- to 10-membered heterocycloalkyl group, where the substituents on the cycloalkyl or heterocycloalkyl group are selected from one or more of a halogen atom, an amino group, a hydroxyl group, a nitro group, a cyano group, a mercapto group, an optionally substituted C1-8 alkyl group, an optionally substituted C1-8 alkoxy group, an optionally substituted C1-8 alkylthio group, and an optionally substituted C1-8 alkylamino group;

[0020] The substituents on the C1-8 alkyl group are selected from one or more of a halogen atom, an amino group, a hydroxyl group, a nitro group, a cyano group, and a mercapto group;

[0021] n is 0, 1, or 2, and m is 0, 1, 2, 3, 4, or 5.

[0022] In one set of embodiments, R5 is an optionally substituted 3- to 10-membered cycloalkyl group, preferably an optionally substituted 3- to 6-membered cycloalkyl group, more preferably a 3- to 6-membered cycloalkyl group substituted with a halogen atom, and even more preferably R5 is a 3- to 6-membered cycloalkyl group substituted with fluorine.

[0023] Furthermore, R5 is Preferably, R5 is

[0024] In one set of embodiments, R6 is a methyl group.

[0025] In one set of embodiments, R2 is H.

[0026] In one set of embodiments, B is an optionally substituted 3- to 6-membered heterocycloalkyl group, preferably an optionally substituted 5- to 6-membered heterocycloalkyl group, and more preferably an optionally substituted piperidinyl group or an optionally substituted piperazinyl group.

[0027] In one set of embodiments, the compound represented by Formula I is Formula I-

[0028]

[0029] R 10 is selected from a halogen atom, an amino group, a hydroxyl group, a nitro group, a cyano group, a mercapto group, an optionally substituted C1-8 alkyl group, an optionally substituted C1-8 alkoxy group, an optionally substituted C1-8 alkylthio group, and an optionally substituted C1-8 alkylamino group;

[0030] R 11is H or an optionally substituted C1-8 alkyl group;

[0031] The substituents on the C1-8 alkyl group are selected from one or more of halogen, amino, hydroxy, nitro, cyano, and mercapto;

[0032] q is 0, 1, 2, 3, or 4.

[0033] Further, R 11 is H or methyl.

[0034] In one set of embodiments, A is an optionally substituted phenyl group or an optionally substituted 5-membered heteroaryl group.

[0035] Further, A is one of the following optionally substituted groups:

[0036]

[0037]

[0038] Preferably, it is connected to the amino group above and to the B group below.

[0039] In one set of embodiments, R3 is chlorine, preferably substituted at the para position of the amino group.

[0040] In one set of embodiments, the substituent on the A ring is methoxy, preferably substituted at the ortho position of the amino group. In one set of embodiments, the compound represented by the formula I is one of the following compounds:

[0041]

[0042]

[0043] In one set of embodiments, the pharmaceutically acceptable salt is formate.

[0044] In one set of embodiments, the isotope derivative is a deuterated compound.

[0045] On the other hand, the present application also provides a method for preparing the pyrimidineamine compound, which comprises the following steps: preparing the compound of formula I from the compound of formula II and the compound of formula III;

[0046] or

[0047] preparing the compound of formula V from the compound of formula IV and the compound of formula II, and then preparing the compound of formula I from the compound of formula V;

[0048] Optionally, necessary protection and deprotection steps are included in the preparation process;

[0049]

[0050]

[0051] Wherein, Z is a leaving group, preferably Z is a halogen, more preferably Z is Cl, and the definitions of the other groups are as defined above.

[0052] In a set of embodiments, the amino or imino group on the B ring in formula II is first protected, and after coupling with the compound of formula III, the protecting group is then removed to prepare the compound of formula I.

[0053] In a set of embodiments, the amino or imino group connected to R2 in the compound of formula IV is first protected, and optionally the amino or imino group on the B ring in formula II is protected. Then, the protected compound of formula IV is coupled with the optionally protected compound of formula II, and the protecting group on the amino or imino group connected to R2 is removed to prepare the compound of formula V. Then, the compound of formula V reacts with R5-Y-OH, and optionally the protecting group on the B ring is removed to prepare the compound of formula I.

[0054] 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.

[0055] The present application provides the use of the pyrimidineamine compound for preparing a NUAK1 or NUAK2 inhibitor.

[0056] The present application provides the use of the pyrimidineamine compound for preparing a drug, 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, skin fibrosis diseases.

[0057] In a set of embodiments, the diseases are Parkinson's disease, Alzheimer's disease, diabetes, hyperlipidemia, obesity, liver cancer, leukemia, lymphoma, tumor metastasis, liver cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or used alone to reduce trauma and postoperative scars.

[0058] The beneficial effects of the present invention are:

[0059] The present invention provides a class of pyrimidineamine compounds. In vitro kinase activity inhibition tests show that the compounds of the present invention have excellent inhibitory activity against NUAK1 / NUAK2 kinases. They can be used for preventing or treating the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, skin fibrosis diseases. Detailed Embodiments

[0060] The following provides a detailed description of the specific embodiments of the present invention. It should be understood that the specific embodiments described herein are only for the purpose of illustrating and explaining the present invention, and are not intended to limit the present invention.

[0061] In the ranges disclosed herein, the endpoints and any values are not limited to the exact ranges or values. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values, they can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0062] Before describing the present invention in detail, it should be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of the present invention. The scope of the present invention is only defined by the appended claims. To more fully understand the present invention described herein, the following terms are used and their definitions are shown below. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains.

[0063] Definitions

[0064] Unless otherwise specified, the following terms referred to in the present invention have the following definitions.

[0065] Features described or depicted as part of one embodiment or a group of embodiments can be used in another embodiment or another group of embodiments to yield further embodiments.

[0066] In the present invention, on the ring group It means that there can be n R3s, m R4s, q Rs 10 Connected at any possible position on the said ring group.

[0067] In the present invention, in some substituents The position indicated represents the connection site.

[0068] Unless otherwise specified, the term "optionally substituted" means that the hydrogen on the group to be substituted is either unsubstituted or one or more of the substitutable sites of the substituent group are independently substituted by substituents, and the substituents are independently selected from one or more of deuterium, halogen, amino, hydroxy, nitro, cyano, mercapto, oxo, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino; the substituents on C1-8 alkyl are selected from one or more of deuterium, halogen, amino, hydroxy, nitro, cyano, mercapto; when the substituent is selected from "oxo", it means that two hydrogen atoms at the same substitution position are replaced by an oxygen atom.

[0069] The term "aryl" refers to a monocyclic or bicyclic aromatic carbocyclic system containing 6 to 10 carbon atoms. Examples of aryl groups include phenyl and naphthyl.

[0070] The term "heteroaryl" refers to an aromatic monocyclic or polycyclic ring system having a 5- to 10-membered structure, or preferably a 5- to 8-membered structure, more preferably a 5- to 6-membered structure, in which one, two, three, four 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 can be an aromatic monocyclic ring having a 5- to 6-membered structure containing 1 to 2 heteroatoms selected from N or S, or an aromatic monocyclic ring having a 5- to 6-membered structure containing 1 to 2 N atoms. Examples of heteroaryl include, but are not limited to, furyl, thienyl, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiodiazolyl, triazinyl, phthalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purinyl, indolyl, isoindolyl, indazolyl, benzofuryl, benzothienyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, benzimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridyl, imidazo[1,2-a]pyridyl, pyrazolo[1,5-a]pyridyl, 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]pyridyl, etc.

[0071] The term "cycloalkyl" refers to a saturated monocyclic, bicyclic or tricyclic system containing 3 to 10 carbon atoms, in which the monocyclic, bicyclic or tricyclic ring does not contain an aromatic ring. The bicyclic group includes a bridged ring group, a spiro ring group, a fused ring group, etc. Preferably, it contains 3 to 10 carbon atoms (C3-10 cycloalkyl), more preferably 3 to 8 carbon atoms (C3-8 cycloalkyl), 3 to 6 carbon atoms (C3-6 cycloalkyl), 4 to 6 carbon atoms (C4-6 cycloalkyl), 5 to 6 carbon atoms (C5-6 cycloalkyl). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, etc.

[0072] The term "heterocycloalkyl" refers to a saturated monocyclic, bicyclic or polycyclic cyclic hydrocarbon group, preferably containing 3-10 ring atoms, where 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, fused ring groups, etc. 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 heteroatoms are preferably 1-4, more preferably 1-3 (i.e., 1, 2 or 3). The heterocycloalkyl can be a 5-6 membered monocyclic heterocycloalkyl containing 1-2 N. Examples of heterocycloalkyl include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, pyranyl, aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, oxacyclohexane, morpholinyl, thiomorpholinyl, dioxolanyl, dithiane, oxazolidinyl, thiazolidinyl, pyrazolidinyl, imidazolidine, etc.

[0073] The term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, preferably a straight-chain or branched-chain group containing 1-8 carbon atoms (C1-8 alkyl) (the number of carbon atoms is between 1 and 8, specifically 1, 2, 3, 4, 5, 6, 7 or 8), more preferably containing 1-6 carbon atoms (i.e., C1-6 alkyl, the number of carbon atoms is between 1 and 6, 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, etc.

[0074] The terms "alkyloxy", "alkylthio", "alkylamino" refer to -O-alkyl, -S-alkyl, -NH-alkyl or dialkylamino respectively, where the alkyl is as defined above. Representative examples include but are not limited to methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, tert-butoxy, etc.; methylthio, ethylthio, propylthio, isopropylthio, butylthio, 1-methylpropylthio, 2-methylpropylthio, tert-butylthio, etc.; methylamino, ethylamino, propylamino, dimethylamino, diethylamino, dipropylamino, methylethylamino, etc.

[0075] The term "halogen" refers to F, Cl, Br, I.

[0076] The active compounds described in the present invention are construed to include the compounds and their stereoisomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs or pharmaceutically acceptable salts thereof. The stereoisomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs, isotopic derivatives or pharmaceutically acceptable salts of the compounds are obtained by conventional techniques in the art and exert the same or similar effects in vivo and in vitro with substantially the same mechanism of action as the compounds.

[0077] The term "stereoisomer" refers to isomers resulting from different arrangements of atoms in space within a molecule, including configurational isomers and conformational isomers, where configurational isomers include geometric isomers (or cis-trans isomers) and optical isomers (including enantiomers and diastereomers). Geometric isomers may be present in the compounds of the present invention. Optical isomers refer to substances with identical molecular structures, similar physicochemical properties, but different optical activities. The compounds of the present invention may contain asymmetrically substituted carbon atoms in the R or S configuration, where 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 (with an equal number of R and S configurations) are racemic at those carbon atoms. An excess of one configuration (relative to the other) of atoms results in a higher number of that configuration, preferably in an excess of about 85%-90%, more preferably in an excess of about 95%-99%, and even more preferably in an excess greater than about 99%. Accordingly, the present invention includes racemic mixtures, relative and absolute optical isomers, and mixtures of relative and absolute optical isomers.

[0078] The term "tautomer" refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also known as proton-transfer tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some bonding electrons.

[0079] The term "isotopic derivative" means that the compounds of the present invention may exist in isotopically labeled or enriched forms, containing one or more atoms whose atomic weights or mass numbers are different from those of the atoms found in the greatest abundance in nature. Isotopes may be radioactive or non-radioactive isotopes. Isotopes of atoms such as hydrogen, carbon, phosphorus, sulfur, fluorine, chlorine, and iodine include, but are not limited to: 2 H, 3H, 13 C, 14 C, 15 N, 18 O, 32 P, 35 S, 18 F, 36 Cl and 125 I. Compounds containing these and / or other isotopes of these atoms are within the scope of the present invention. The isotopically labeled compounds of the present invention can be prepared by general methods well known to those of ordinary skill in the art.

[0080] The term "hydrate" refers to an association formed by one or more water molecules with the compounds of the present invention.

[0081] The term "solvate" refers to an association formed by one or more solvent molecules with the compounds of the present invention.

[0082] The term "prodrug" is a derivative of a designed active drug that can improve certain undesirable physical or biological properties. Physical properties are usually related to solubility (excessive or insufficient lipid or water solubility) or stability, while problematic biological characteristics include too rapid metabolism or poor bioavailability, which may itself be related to physicochemical properties.

[0083] The term "pharmaceutically acceptable salt" refers to a salt that is suitable for contact with mammalian, particularly human, tissues within the scope of reasonable medical judgment, without excessive toxicity, irritation, allergic reaction, etc., and commensurate with a reasonable benefit / risk ratio. 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.

[0084] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent, or emulsifying agent, etc., that has been approved by the relevant government regulatory authorities for use in humans or livestock.

[0085] As used herein, the term "treatment" refers to any administration of a therapeutic agent according to a therapeutic regimen that achieves a desired effect, namely, partial or complete alleviation, amelioration, remission, inhibition, delay in onset, reduction in severity, and / or reduction in 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 can be directed to a subject who does not exhibit the relevant disease, disorder, and / or condition and / or to a subject who exhibits only early signs of the disease, disorder, and / or condition. Alternatively or additionally, such treatment can be directed to a subject who exhibits one or more identified signs of the relevant disease, disorder, and / or condition. In some embodiments, the treatment can be directed to a subject who has been diagnosed with the relevant disease, disorder, and / or condition. In some embodiments, the treatment can be directed to a subject known to have one or more risk factors that are statistically associated with an increased risk of developing the relevant disease, disorder, and / or condition.

[0086] According to the present invention, the drug prepared in the pharmaceutical use described in the present application may contain, in addition to the pyrimidineamine compound of the present invention as an active ingredient, other agents that can be used to prevent or treat related diseases as another active ingredient. When the drug contains multiple active ingredients, the active ingredients can be administered simultaneously, sequentially, or separately according to the judgment of a physician.

[0087] Hereinafter, the effects of specific compounds of the present invention will be described in detail by way of examples.

[0088] Example

[0089] General method for synthesizing compound 537 (TDM-181137) in Example 1

[0090]

[0091] Step 1: Compound 537c

[0092] tert-Butyl 1-(2-chloropyrimidin-4-yl)-4-methylpiperidin-4-ylcarbamate

[0093] 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). The reaction solution was stirred at 50 °C 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 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.

[0094] Step 2: Compound 537d

[0095] 4-chloropyrimidine-4-chloropyridin-4-amine

[0096] To a solution of compound 537c (548.3 mg, 1.68 mmol) in dichloromethane (22 mL) was added trifluoroacetic acid (2.49 mL). 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 pH was adjusted to basic with potassium carbonate solution. Then, the mixture was extracted with ethyl acetate (3 × 50 ml). 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). The crude product was directly used in the subsequent step. LCMS [M+1] + = 227.1.

[0097] Step 3: Compound 537f

[0098] N-(1-(2-chloropyrimidin-4-yl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0099] To a solution of compound 537d (337.2 mg, 1.49 mmol) in N,N-dimethylformamide (15 mL) was 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 h. After completion of 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), dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0 - 44 / 56) to give a white solid compound (compound 537f, 231.8 mg, yield 47%), LCMS [M+1] + = 331.1.

[0100] Step 4: Compound 535h

[0101] tert-Butyl 4-(4-(4-(4-cyclopropanesulfonamido)-4-methylpiperidin-1-yl)pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0102] 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) was 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, purged with argon, and the reaction solution was stirred at 100 °C for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure. 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.

[0103] Step 5: Compound 535

[0104] N-(4-Methyl-1-(2-(1-(4-piperidinyl)-1H-pyrazol-4-ylamino)pyrimidin-4-yl)piperidin-4-yl)cyclopropanesulfonamide

[0105] 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. After completion of the reaction, the mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to obtain a white solid compound (compound 537, 18.9 mg, 23% yield). LCMS [M+1] + = 461.2.

[0106] 1 H NMR (400 MHz, MeOD) δ 7.93 (s, 1H), 7.66 (s, 2H), 6.58 (d, J = 7.3 Hz, 1H), 4.59 (s, 2H), 3.91 (s, 1H), 3.66 (s, 2H), 3.57 (d, J = 12.7 Hz, 2H), 3.23 (t, J = 10.8 Hz, 2H), 2.63 (dq, J = 7.9, 4.9 Hz, 1H), 2.33 (s, 4H), 2.19 (d, J = 13.9 Hz, 2H), 1.65 (t, J = 10.6 Hz, 2H), 1.52 (s, 3H), 1.15–1.10 (m, 2H), 1.09–1.03 (m, 2H).

[0107] Example 2: General method for synthesizing compound 563 (TDM-181163)

[0108]

[0109] Step 1: Compound 563c

[0110] (1-(2-Chloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamic acid tert-butyl ester

[0111] To a solution of compound 563a (300 mg, 2.014 mmol) and compound 563b (453 mg, 2.115 mmol) in acetonitrile (30 mL) was added potassium carbonate (557 mg, 4.028 mmol), and the mixture was heated to 50 °C and stirred overnight. The mixture was added to water (150 mL), extracted with ethyl acetate (50 mL * 3), the combined organic layers were 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 obtain a white solid product (compound 563c, 528.5 mg, 80.3% yield). LCMS [M+1] + = 327.

[0112] Step 2: Compound 563e

[0113] Benzyl 4-(4-((4-(4-(tert-Butoxycarbonyl)amino)-4-methylpiperidin-1-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0114] To a mixture of compound 563c (109 mg, 0.333 mmol), compound 563d (100 mg, 0.333 mmol), 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, purged with argon several times, then heated to 100 °C and stirred for 2 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 62 / 38 with 10% methanol) to give the white solid product (compound 563e, 131.3 mg, 66.7% yield). LCMS [M+1] + = 591.

[0115] Step 3: Compound 563f

[0116] Benzyl 4-(4-((4-(4-Amino-4-methylpiperidin-1-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0117] To a solution of compound 563e (90 mg, 0.152 mmol) in dichloromethane (9 mL) at room temperature was added trifluoroacetic acid (0.7 mL). The mixture was stirred for 1 h. Then the mixture was concentrated under reduced pressure, water was added to the residue and neutralized with aqueous sodium carbonate solution, extracted with ethyl acetate (20 mL * 3), the combined organic layers were washed with brine, and the filtrate was concentrated under reduced pressure to give the yellow solid product (compound 563f, 73.5 mg, 98.5% yield). LCMS [M+1] + = 491.

[0118] Step 4: Compound 563h

[0119] (S)-Benzyl 4-(4-((4-(4-(2,2-Difluorocyclopropane-1-carboxamido)-4-methylpiperidin-1-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0120] 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) at room temperature was added 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (85 mg, 0.22 mmol) and compound 563f (27.5 mg, 0.225 mmol). The mixture was stirred for 1 hour. Then the mixture was diluted with water and extracted with ethyl acetate (30 mL * 3). The combined organic layers were washed with brine, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 70 / 30 with 10% methanol) to give a yellow solid product (compound 563h, 80 mg, 61.1% yield). LCMS [M+1] + = 595.

[0121] Step 5: Compound 563

[0122] (S)-2,2-difluoro-N-(4-methyl-1-(2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)piperidin-4-yl)cyclopropane-1-carboxamide

[0123] 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. Then the mixture was 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.

[0124] 1 H NMR (400 MHz, DMSO) δ 8.84 (s, 1H), 8.18 (s, 1H), 7.99 (s, 1H), 7.89 (d, J = 6.0 Hz, 1H), 7.80 (s, 1H), 7.51 (s, 1H), 6.19 (d, J = 6.1 Hz, 1H), 4.39 (dt, J = 14.9, 5.4 Hz, 1H), 3.90 (s, 2H), 3.36 (d, J = 12.8 Hz, 2H), 3.26 (s, 3H), 3.05–2.97 (m, 2H), 2.67 (ddd, J = 14.7, 10.9, 8.0 Hz, 1H), 2.13 (d, J = 13.7 Hz, 4H), 2.09–1.97 (m, 2H), 1.90–1.72 (m, 2H), 1.49 (d, J = 11.1 Hz, 2H), 1.32 (s, 3H).

[0125] Example 3: General method for synthesizing compound 564 (TDM-181164)

[0126]

[0127] Step 1: Compound 564

[0128] (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

[0129] At room temperature, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (44.8 mg, 0.118 mmol) and compound 564b (19.2 mg, 0.157 mmol) were added 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). 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

[0130] 1 H NMR (400 MHz, DMSO) δ 8.75 (s, 1H), 8.16 (s, 2H), 7.97 (s, 1H), 7.89 (d, J = 6.0 Hz, 1H), 7.53 (d, J = 9.0 Hz, 2H), 6.84 (d, J = 9.1 Hz, 2H), 6.21 (d, J = 6.1 Hz, 1H), 3.92 (s, 2H), 3.23 (dt, J = 13.7, 9.0 Hz, 2H), 3.10–3.02 (m, 4H), 2.66 (ddd, J = 14.4, 11.0, 8.1 Hz, 1H), 2.57–2.52 (m, 4H), 2.28 (s, 3H), 2.13 (d, J = 10.6 Hz, 2H), 1.91–1.71 (m, 2H), 1.46 (td, J = 10.5, 4.0 Hz, 2H), 1.31 (s, 3H)

[0131] Example 4: General method for synthesizing compound 565 (TDM-181165)

[0132]

[0133] Step 1: Compound 565

[0134] N-(4-Methyl-1-(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)piperidin-4-yl)cyclopropanesulfonamide

[0135] At room temperature, 4-dimethylaminopyridine (9.6 mg, 0.079 mmol) and compound 565b (19.2 mg, 0.157 mmol) were added 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). The mixture was heated to 40 °C and stirred for 3 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 75 / 25 with 10% methanol) and preparative HPLC (formic acid) to give the white solid product (compound 565, 13.3 mg, 34.7% yield). LCMS [M+1] + = 486.3.

[0136] 1 H NMR (400 MHz, DMSO) δ 8.74 (s, 1H), 8.14 (s, 2H), 7.89 (d, J = 6.1 Hz, 1H), 7.53 (d, J = 9.0 Hz, 2H), 6.86 (d, J = 4.1 Hz, 2H), 6.84 (s, 1H), 6.21 (d, J = 6.1 Hz, 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 = 14.1 Hz, 2H), 1.57–1.46 (m, 2H), 1.40 (s, 3H), 0.97 (d, J = 3.4 Hz, 2H), 0.95 (s, 2H).

[0137] Example 5: General method for the synthesis of compound 572 (TDM-181172)

[0138]

[0139] Step 1: Compound 572c

[0140] (1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamate

[0141] 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, by 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, the combined organic phase was 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), eluting with PE / EA = 0 - 60%, to give a white solid (Compound 572c, 1.65 g, 84% yield) as the product. LCMS [M+1] + = 361.2.

[0142] Step 2: Compound 572d

[0143] 1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-amine

[0144] 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 then the mixture was stirred at room temperature for 1 hour. The product was detected by LCMS [M+1] + = 261.1. The mixture was added to water (60 mL), extracted with ethyl acetate (2 * 60 mL), and the organic layers were combined. The combined organic phase was 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), eluting with DCM / (DCM:MeOH = 10:1) = 0 - 50%, to give a yellow solid (Compound 572d, 870 mg, crude) as the product. LCMS [M+1] + = 261.1.

[0145] Step 3: Compound 572f

[0146] (S)-N-(1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0147] To a 50 mL flask was added a solution of Compound 572d (350 mg, 1.3� 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), N,N-Diisopropylethylamine (261 mg, 2.01 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The product was detected by LCMS [M+1] +Product was detected at m / z = 365.1. The combined organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrated organic layer was purified by flash chromatography (40 g column, over 35 min) eluting with PE / EA = 0 - 75% to give a yellow oil (Compound 572f, 670 mg, crude) as the product. LCMS [M+1]+ = 365.1.

[0148] Step 4: Compound 572

[0149] (S)-N-(1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0150] To a 50 mL beaker was added Compound 572f (74.5 mg, 0.2 mmol), Compound 572e (45 mg, 0.2 mmol), and p-toluenesulfonic acid (77 mg, 0.407 mmol) dissolved in n-butanol (4 mL), and the mixture was stirred at 110 °C for 4 h. Product was detected by LCMS [M+1] + = 550.3. The mixture was added to water (20 mL) and extracted with ethyl acetate (2 x 20 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give a white solid product (Compound 572, 5.3 mg, 13% yield). LCMS [M+1] + = 550.3.

[0151] 1 H NMR (401 MHz, DMSO) δ 8.20 (s, 1H), 7.98 (s, 1H), 7.95 (s, 1H), 7.72–7.67 (m, 2H), 6.61 (d, J = 2.5 Hz, 1H), 6.45 (dd, J = 8.9, 2.5 Hz, 1H), 3.84 (d, J = 14.4 Hz, 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 = 13.0 Hz, 2H), 1.88–1.72 (m, 2H), 1.56 (dd, J = 13.8, 10.4 Hz, 2H), 1.32 (s, 3H).

[0152] N-(1-(5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide (TDM-181173), a white solid (52.2 mg, yield 37.17%), and (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), a white solid (5 mg, yield 60%) were synthesized by a method similar to that of Example 5.

[0153]

[0154]

[0155] Example 6: General method for preparing Compound 575 (TDM-181175)

[0156]

[0157] Step 1: Compound 575c

[0158] (1-(2,5-dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamate

[0159] 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 overnight at 50 °C, and the product was detected by LCMS [M+1] + = 361.2. The mixture was added to water (50 ml) and extracted with ethyl acetate (2 × 50 mL). The combined organic phases were 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), eluting with PE / EA = 0 - 60% to give the product as a white solid (Compound 575c, 1.65 g, yield 84%). LCMS [M+1] + = 361.2.

[0160] Step 2: Compound 575d

[0161] 1-(2,5-dichloropyrimidin-4-yl)-4-methylpiperidin-4-amine

[0162] The hydrochloric acid - dioxane solution (10 mL, 134.62 mmol) was added to a dioxane (100 mL) solution containing compound 575c (1.1 g, 3.05 mmol) at 0 °C, and then the mixture was stirred at room temperature for 1 hour. The product was detected by LCMS [M+1] + = 261.1. The mixture was added with water (60 mL), extracted with ethyl acetate (2×60 mL), and the organic layers were combined. The combined organic phase was 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), eluted with DCM / (DCM:MeOH = 10:1) = 0 - 50%, and a yellow solid (compound 575d, 870 mg, crude) was obtained as the product. LCMS [M+1] + = 261.1.

[0163] Step 3: Compound 575f

[0164] N-(1-(2,5 - Dichloropyrimidin - 4 - yl)-4 - methylpiperidin - 4 - yl)cyclopropanesulfonamide

[0165] 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), and 4 - dimethylaminopyridine (20,243 mg, 1.66 mmol) and compound 575e (349.40 mg, 2.49 mmol) were added at 0 °C. The reaction mixture was heated to 40 °C and stirred for 3 hours. LCMS [M+1] + = 365, and the reaction was detected to be complete. Workup: The reaction mixture was poured into water, and the aqueous phase was extracted with dichloromethane (2×40 mL). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness, and the resulting crude product was purified by column chromatography [eluent: EA / PE = 0 - 30%] to obtain the target compound as a yellow liquid (compound 575f, 554.9 mg, yield 82%), LCMS [M+1] + = 365.

[0166] Step 4: Compound 575

[0167] N-(1-(5 - Chloro - 2 - ((2 - methoxy - 4-(4 - methylpiperazin - 1 - yl)phenyl)amino)pyrimidin - 4 - yl)-4 - methylpiperidin - 4 - yl)cyclopropanesulfonamide

[0168] 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 mixture was purged with argon several times and then anhydrous 1,4-dioxane (10 mL) was added. The reaction mixture was purged with argon three times. The temperature was raised to 100 °C and stirred for two hours. LCMS [M+1] + = 550.2, indicating that the reaction was complete. Work-up: The crude product obtained by concentrating the reaction mixture to dryness was purified by column chromatography [eluent: (dichloromethane / methanol = 10:1) / DCM = 0 - 95%]. The crude product was further purified by preparative purification to obtain the white solid target compound (compound 575, 75.7 mg, yield 33.56%), LCMS [M+1] + = 550.2.

[0169] 1 H NMR (400 MHz, MeOD) δ 8.44 (s, 1H), 8.01 (d, J = 8.8 Hz, 1H), 7.91 (d, J = 1.7 Hz, 1H), 6.68 (d, J = 2.3 Hz, 1H), 6.56 (dd, J = 8.8, 2.3 Hz, 1H), 3.97 (d, J = 13.1 Hz, 2H), 3.89 (s, 3H), 3.58 (t, J = 11.8 Hz, 2H), 3.28 (d, J = 5.3 Hz, 4H), 3.03 (d, J = 4.7 Hz, 4H), 2.66 (d, J = 2.7 Hz, 3H), 2.63–2.54 (m, 1H), 2.09 (d, J = 13.9 Hz, 2H), 1.74–1.65 (m, 2H), 1.50 (s, 3H), 1.13–1.07 (m, 2H), 1.06–0.98 (m, 2H).

[0170] Example 7: General method for the synthesis of compound 578 (TDM-181178)

[0171]

[0172] Step 1: Compound 578c

[0173] (1-(2,5-Dichloropyrimidin-4-yl)-4-methylpiperidin-4-yl)carbamate

[0174] To a solution of compound 578a (1 g, 5.45 mmol) and compound 578b (1.17 g, 5.45 mmol) in acetonitrile (50 mL) was added potassium carbonate (1.5 g, 10.9 mmol). The reaction solution was stirred overnight at 50 °C, and the product was detected by LCMS [M+1] + = 361.2. The mixture was added to water (50 ml) and extracted with ethyl acetate (2 * 50 mL). The organic phases were combined, the combined organic phase was 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), eluting with PE / EA = 0 - 60%, to give a white solid (compound 578c, 1.65 g, 84% yield) as the product. LCMS [M+1] + = 361.2.

[0175] Step 2: Compound 578e

[0176] Benzyl 4-(4-((4-(4-(tert-butoxycarbonyl)amino)-4-methylpiperidin-1-yl)-5-chloropyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0177] A 50 mL flask was charged with 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(diphenylphosphino)-9,9-dimethyloxanthrene (161.26 mg, 0.279 mmol) and cesium carbonate (454 mg, 1.4 mmol) and dissolved in ultradry 1,4-dioxane (25 mL), and the mixture was stirred under argon at 100 °C for 2 hours. The reaction was monitored by TLC and LCMS. The product was confirmed by LCMS [M+1] + = 625.3. The mixture was added to water (50 mL), extracted with ethyl acetate (2 * 50 mL), and the organic layers were combined. The combined organic phase was 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 give a white solid (compound 578e, 240 mg, 50% yield) as the product LCMS [M+1] + = 625.3.

[0178] Step 3: Compound 578f

[0179] Benzyl 4-(4-((4-(4-amino-4-methylpiperidin-1-yl)-5-chloropyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0180] Dissolve compound 578e (100 mg, 0.16 mmol) in dichloromethane (10 mL) at 0 °C, add trifluoroacetic acid (1 mL), and then stir the mixture at room temperature for 1 hour. Monitor the reaction by TLC and LCMS. Detect the product by LCMS [M+1] + = 525.3. Concentrate the reaction mixture under reduced pressure, pour it into water (30 mL), and adjust the pH > 8 with potassium carbonate (50%). Add water (30 ml) and ethyl acetate (2 * 30 ml) to extract the mixture, and combine the organic layers. Wash the combined organic phase with brine, dry it over anhydrous sodium sulfate, and concentrate it under reduced pressure to obtain a yellow solid as the product (compound 578f, 170 mg, crude). LCMS [M+1] + = 525.3.

[0181] Step 4: Compound 578h

[0182] 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

[0183] Take a 50 mL flask, dissolve compound 57,8f (14 mg, 0.27 mmol), compound 578g (48.8 mg, 0.4 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (152 mg, 0.4 mmol) in N,N-dimethylformamide (15 mL), add N,N-diisopropylethylamine (51.6 mg, 0.4 mmol), and stir the mixture at room temperature for 1 hour. Monitor the reaction by TLC and LCMS. Confirm the product by LCMS [M+1]+ = 629.2. Add water (30 mL) to the mixture, extract it with ethyl acetate (2 * 30 mL), and combine the organic layers. Wash the combined organic phase with brine, dry it over anhydrous sodium sulfate, and concentrate it under reduced pressure. Concentrate the organic layer and purify it by flash chromatography (12 g column, 25 minutes), eluting with DCM / (DCM:MeOH = 10:1) = 0 - 100% to obtain the target product as a yellow oil (compound 578h, 133 mg, 74% yield). LCMS [M+1] + = 629.2.

[0184] Step 5: Compound 578

[0185] (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-yl)cyclopropane-1-carboxamide

[0186] Take a 25 mL flask, add compound 578h (40 mg, 0.064 mmol) at 0 °C and cool for a period of time. Then, dropwise add acetic acid solution of hydrobromic acid (2 mL). The mixture is stirred at 0 °C for 0.5 h. Detect the reaction by TLC and LCMS. The product is detected by LCMS [M+1] + = 495.2. Concentrate the mixture under reduced pressure. Purify the crude product by preparative HPLC to obtain a white solid product (compound 578, 12 mg, yield 37.5%), LCMS [M+1] + = 495.2.

[0187] 1 H NMR (401 MHz, DMSO) δ 9.24 (s, 1H), 8.29 (s, 1H), 8.01 (s, 2H), 7.78 (s, 1H), 7.52 (s, 1H), 4.26 (s, 1H), 3.85 (d, J = 8.6 Hz, 2H), 3.31 (s, 2H), 3.20 (d, J = 12.0 Hz, 2H), 2.80 (t, J = 11.5 Hz, 2H), 2.72–2.63 (m, 1H), 2.19 (d, J = 11.3 Hz, 2H), 2.03 (d, J = 11.8 Hz, 2H), 1.97–1.64 (m, 5H), 1.60 (t, J = 11.9 Hz, 2H), 1.33 (s, 3H).

[0188] Example 8: General method for synthesizing compound 582 (TDM-181182)

[0189]

[0190] Step 1: Compound 582c

[0191] 2,5-Dichloro-4-vinylpyrimidine

[0192] 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(diphenylphosphino)ferrocene dichloropalladium(0) (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 h. The reaction was monitored and found to be complete. Workup: The reaction mixture was poured into water and the aqueous layer was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and dried under reduced pressure. The crude product was purified by column chromatography [eluent: EA / PE = 0 - 5%] to afford the title compound as a yellow oil (compound 582c, 960 mg, 47% yield). LCMS [M+1] + = 175, 177.

[0193] Step 2: Compound 582d

[0194] 2,5-Dichloropyrimidine-4-carbaldehyde

[0195] Compound 582c (700 mg, 60% purity) was dissolved in dichloromethane (15 mL). The solution was cooled to -78 °C and purged with ozone for 30 min. Then dimethyl sulfide was added and the mixture was stirred for 3 min. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (ethyl acetate / petroleum ether = 0 - 30 / 70) to afford the product as a white solid (compound 582d, 323 mg, crude). LCMS [M+1] + = 177&179.

[0196] Step 3: Compound 582f

[0197] tert-Butyl (1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0198] To a solution of compound 582d (323 mg, 1.825 mmol) in dichloromethane (18 mL) at room temperature were added compound 582e (430 mg, 2.008 mmol) and acetic acid (2 drops). The mixture was stirred for 1 h and sodium triacetoxyborohydride (580 mg, 2.738 mmol) was added. The mixture was stirred for 20 min and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (dichloromethane / methanol in dichloromethane with 10% methanol = 0 - 15 / 85) to afford the product as a white solid (compound 582f, 350 mg, 51.1% yield). LCMS [M+1] + = 375&377.

[0199] Step 4: Compound 582h

[0200] tert-Butyl (1-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0201] At room temperature, dioxane (10 mL) was added 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). Then 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 (dichloromethane / methanol = 0 - 80 / 20 with 10% methanol) to give a white solid product (compound 582h, 125 mg, 69.1% yield). LCMS [M+1] + = 530.

[0202] Step 5: Compound 582i

[0203] 4-((4-Amino-4-methylpiperidin-1-yl)methyl)-5-chloro-N-(4-(4-methylpiperazin-1-yl)phenyl)pyrimidin-2-amine

[0204] At room temperature, trifluoroacetic acid (1.5 mL) was added to a solution of compound 582h (125 mg, 0.236 mmol) in dichloromethane (10 mL). Then the mixture was stirred at room temperature for 1 hour. The mixture was diluted with water and neutralized with aqueous potassium carbonate solution, extracted with ethyl acetate, the combined organic layers were dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was used directly in the next reaction. LCMS [M+1] + = 430.

[0205] Step 6: Compound 582

[0206] (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

[0207] At room temperature, 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-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (66 mg, 0.174 mmol) and N,N-diisopropylethylamine (22.5 mg, 0.174 mmol), and then the mixture was stirred for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (dichloromethane / methanol = 100 / 0 to 0 / 100 with 10% methanol) and preparative HPLC (formic acid) to give the yellow solid product (compound 582, TDM-181182, 8.8 mg, 14.2% yield). LCMS [M+1] + = 534.

[0208] 1 H NMR (400 MHz, DMSO) δ 10.44 (s, 1H), 9.74 (s, 1H), 8.49 (s, 1H), 8.14 (s, 1H), 7.57 (d, J = 7.9 Hz, 2H), 6.97 (d, J = 8.8 Hz, 2H), 4.57 (s, 2H), 3.68 (s, 4H), 3.45 (s, 2H), 2.99 (s, 6H), 2.82 (s, 3H), 2.65 (d, J = 12.9 Hz, 2H), 2.33 (s, 2H), 1.83 (d, J = 6.7 Hz, 4H), 1.31 (s, 3H).

[0209] Example 9: General method for the synthesis of compound 583 (TDM-181183)

[0210]

[0211] Step 1: Compound 583c

[0212] 2,5-Dichloro-4-vinylpyrimidine

[0213] 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(diphenylphosphino)ferrocene dichloropalladium (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, heated to 70 °C and reacted for 1 hour. The reaction was monitored and found to be complete. Work-up: The reaction mixture was poured into water, and the aqueous phase was extracted with ethyl acetate (3 × 50 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and dried to obtain a crude product. The crude product was purified by column chromatography [eluent: EA / PE = 0 - 5%] to obtain the target compound as a yellow oil (compound 583c, 960 mg, yield 47%). LCMS [M+1] + = 175, 177.

[0214] Step 2: Compound 583d

[0215] 2,5-Dichloropyrimidine-4-carbaldehyde

[0216] To a solution of compound 583c (960 mg, 5.48 mmol) in dichloromethane (130 mL) in a 100 mL three-necked flask, the reaction mixture was cooled to -78 °C and sufficient ozone was introduced. The reaction was carried out for 15 minutes. The reaction was monitored and found to be complete. Work-up: Dimethyl sulfide was added dropwise at -78 °C, and the reaction mixture was warmed to room temperature and stirred for 30 minutes. The mixture was concentrated and dried to obtain a crude product. The crude product was purified by column chromatography [eluent: EA / PE = 0 - 30%] to obtain the target compound as a yellow oil (compound 583d, 90 mg, yield 13.3%). LCMS [M+1] + = 177, 179.

[0217] Step 3: Compound 583f

[0218] tert-Butyl (1-((2,5-dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0219] To a solution of compound 583d (90 mg, 0.51 mmol) in dichloromethane (10 mL) was added compound 583e (109 mg, 0.51 mmol) and acetic acid (1 drop). The reaction mixture was stirred at room temperature for 1.5 h, then sodium triacetoxyborohydride (162.1 mg, 0.77 mmol) was added, and stirring was continued for 30 min. The reaction was monitored to completion. Workup: The reaction mixture was poured into water, and extracted three times with ethyl acetate (3×50 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and dried to obtain the crude product, which was purified by column chromatography [eluent: EA / PE = 0 - 30%] to give the target compound as a yellow solid (compound 583f, 76 mg, yield 53%). LCMS [M+1] + = 375, 377.

[0220] Step 4: Compound 583g

[0221] 1-((2,5-Dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-amine

[0222] 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 h. The reaction was monitored to completion. Workup: The reaction mixture was poured into water, and extracted with ethyl acetate (1×50 mL) to remove impurities. The aqueous phase was adjusted to pH = 11 with potassium carbonate, and extracted three times with ethyl acetate (3×80 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and dried to obtain the target compound as a yellow solid (compound 583g, 86.5 mg, yield 93.3%). LCMS [M+1] + = 275, 277.

[0223] Step 5: Compound 583i

[0224] N-(1-((2,5-Dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0225] To a solution of 583 g (86.5 mg, 0.31 mmol) of the compound in 8 mL of dichloromethane were added 4-dimethylaminopyridine (38.4 mg, 0.31 mmol), N,N-diisopropylethylamine (81.3 mg, 0.63 mmol) and 583 h (88.4 mg, 0.63 mmol). The reaction mixture was heated to 40 °C and stirred for 3 hours. The reaction was monitored and found to be complete. Work-up: The reaction mixture was poured into water, extracted with ethyl acetate (3 × 50 mL), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and dried under reduced pressure. The crude product was purified by column chromatography [eluent: (dichloromethane:methanol = 10:1) / dichloromethane = 0 - 20%] to obtain the target compound as a yellow solid (compound 583 i, 49.6 mg, yield 34%), LCMS [M+1] + = 379, 381.

[0226] Step 6: Compound 583

[0227] N-(1-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)cyclopropanesulfonamide

[0228] To a 100 mL three-necked flask were added compound 583 i (49.6 mg, 0.13 mmol), compound 583 j (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 mixture was purged with argon several times, heated to 100 °C and stirred for 2 hours. The reaction was monitored and found to be complete. Work-up: The reaction mixture was directly dried under reduced pressure. The crude product was purified by column chromatography [eluent: (dichloromethane:methanol = 10:1 / dichloromethane = 0 - 100%)], and the crude product was further purified by preparative purification to obtain the target compound as a yellow solid (compound 583, 8.6 mg, yield 12.4%), LCMS [M+1] + = 534.2.

[0229] 11H NMR (400 MHz, DMSO-d6) δ 9.57 (s, 1H), 8.38 (s, 1H), 8.16 (s, 2H), 7.58 (d, J = 8.9 Hz, 2H), 6.94–6.83 (m, 2H), 6.68 (s, 1H), 3.53 (s, 2H), 3.06 (t, J = 5.0 Hz, 4H), 2.65–2.58 (m, 2H), 2.58–2.52 (m, 4H), 2.47 (d, J = 4.7 Hz, 4H), 2.24 (s, 3H), 1.91 (d, J = 13.2 Hz, 2H), 1.53 (ddd, J = 12.9, 8.5, 3.6 Hz, 2H), 1.33 (s, 3H), 0.92 (d, J = 6.4 Hz, 4H).

[0230] 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 in a similar manner to Example 9, as a yellow solid (8.8 mg, yield 12.4%).

[0231]

[0232]

[0233] Example 10: General method for the synthesis of compound 585 (TDM-181185)

[0234] [[ID=?]]

[0235] Step 1: Compound 585c

[0236] tert-Butyl (1-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)carbamate

[0237] 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) at room temperature was added dioxane (10 mL). The mixture was 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 (dichloromethane / methanol = 0–80 / 20 with 10% methanol) to give the white solid product (compound 585c, 145 mg, 87.9% yield). LCMS [M+1] It should be noted that there seems to be a mislabeled "ID=?" in the original text which has been maintained as is in the translation.+ = 560.

[0238] Step 2: Compound 585d

[0239] 4 - ((4 - Amino - 4 - methylpiperidin - 1 - yl)methyl)-5 - chloro - N-(2 - methoxy - 4-(4 - methylpiperazin - 1 - yl)phenyl)pyrimidin - 2 - amine

[0240] To a solution of compound 585c (145 mg, 0.315 mmol) in dichloromethane (10 mL) at room temperature was added trifluoroacetic acid (1.5 mL), and then the mixture was stirred at 20 °C for 1 hour. The mixture was diluted with water and neutralized with aqueous potassium carbonate solution, extracted with ethyl acetate, the combined organic layers were dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was used directly in the next reaction. LCMS [M+1] + = 460.

[0241] Step 3: Compound 585

[0242] (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

[0243] To a solution of compound 585d (55 mg, 0.120 mmol) in N,N - dimethylformamide (5 mL) at room temperature 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), and then the mixture was stirred for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 100 / 0 with 10% methanol) and preparative HPLC (formic acid) to give a yellow solid product (compound 585, TDM - 181185, 10 mg, 14.8% yield). LCMS [M+1] + = 564.3.

[0244] 11H NMR (400 MHz, DMSO) δ 8.31 (s, 1H), 8.24 (s, 1H), 8.15 (s, 2H), 7.75 (s, 1H), 7.59 (d, J = 8.7 Hz, 1H), 6.62 (d, J = 2.4 Hz, 1H), 6.47 (dd, J = 8.8, 2.4 Hz, 1H), 3.77 (s, 3H), 3.49 (s, 2H), 3.18–3.11 (m, 4H), 2.67 (td, J = 6.5, 4.4 Hz, 1H), 2.62 (dd, J = 8.8, 5.8 Hz, 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, 6.6 Hz, 2H), 1.25 (s, 3H).

[0245] Example 11: General method for synthesizing compound 586 (TDM-181186)

[0246]

[0247] Step 1: Compound 586b

[0248] 1-((2,5-Dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-amine

[0249] To a solution of compound 586a (116 mg, 0.309 mmol) in dichloromethane (10 mL) at room temperature was added trifluoroacetic acid (2 mL), and the mixture was then stirred for 1 hour. The mixture was diluted with water and neutralized with aqueous sodium hydroxide, 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 the white solid product (compound 586b, 68 mg, 80% yield). LCMS [M+1] + = 275&277.

[0250] Step 2: Compound 586d

[0251] (S)-N-(1-((2,5-Dichloropyrimidin-4-yl)methyl)-4-methylpiperidin-4-yl)-2,2-difluorocyclopropane-1-carboxamide

[0252] At room temperature, to a solution of compound 586b (48 mg, 0.1475 mmol) in N,N-dimethylformamide (5 mL) was added compound 586c (32 mg, 0.262 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (100 mg, 0.262 mmol) and N,N-diisopropylethylamine (34 mg, 0.262 mmol), and then the mixture was stirred for 1 hour. The mixture was added with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 15 / 85 with 10% methanol) to obtain a white solid product (compound 586d, 56.8 mg, 85.6% yield). LCMS [M+1] + = 379&381.

[0253] Step 3: Compound 586f

[0254] 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

[0255] At room temperature, to a solution of compound 586d (47 mg, 0.124 mmol) in dioxane (5 mL) was added compound 586e (41 mg, 0.136 mmol) and p-toluenesulfonic acid monohydrate (47 mg, 0.248 mmol), and then the mixture was heated to 100 °C and stirred for 24 hours. The mixture was added with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 50 / 50 with 10% methanol) to obtain a yellow solid product (compound 586f, 21 mg, 26.3% yield). LCMS [M+1] + = 643.

[0256] Step 4: Compound 586

[0257] (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

[0258] 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 obtain a gray solid product (Compound 586, TDM-181186, 6.7 mg, yield 40.4%). LCMS [M+1] + = 509.

[0259] 1 H NMR (400 MHz, DMSO) δ 9.74 (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 = 10.4 Hz, 1H), 4.10–3.99 (m, 1H), 3.54 (s, 2H), 3.34 (d, J = 12.3 Hz, 2H), 2.98 (t, J = 11.0 Hz, 2H), 2.73–2.66 (m, 1H), 2.65–2.55 (m, 2H), 2.39 (s, 2H), 2.08 (dd, J = 27.2, 12.6 Hz, 6H), 1.88–1.70 (m, 2H), 1.48 (d, J = 8.9 Hz, 2H), 1.26 (s, 3H).

[0260] Test Example: Detection of Enzyme Activity Inhibition of NUAK1 / NUAK2 Kinase

[0261] The effects of the pyrimidineamine small molecules involved in this application on the inhibitory activities of NUAK1 and NUAK2 kinases were tested using a kinase activity test method based on the P81 filter paper binding hot spot technology (Nat Biotechnol. 2011, 29: 1039), which is briefly described as follows:

[0262] The test 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.

[0263] Test Procedure:

[0264] 1. Prepare a 20 μM substrate solution with freshly prepared test buffer. The substrate is the CHKtide polypeptide fragment (Sanchez Y. Science. 1997, 277: 1497-1501);

[0265] 2. Add the kinase and mix gently (final concentration of NUAK1 is 10 nM; final concentration of NUAK2 is 50 nM);

[0266] 3. Using acoustic liquid handling technology (Echo550; nanoliter scale), add the test compound dissolved in 100% DMSO to the above kinase reaction mixture and incubate at room temperature for 5 minutes;

[0267] 4. Add 33 P-labeled ATP (unlabeled ATP / labeled ATP ratio is 25:1 or 2.5:1);

[0268] 5. Incubate at room temperature for 2 hours;

[0269] 6. Detect the kinase activity by P81 filter paper binding hot spot technology.

[0270] Calculate the IC 50 of the test compound against NUAK1 / NUAK2, and the specific results are shown in Table 1.

[0271] IC 50 The calculation uses the formula obtained from a sigmoidal dose-response curve (variable slope):

[0272] Y = Bottom + (Top - Bottom) / (1 + 10^((LogEC50 - X) * slope)), where X is the Log value of the compound concentration, Y is the response (inhibition rate of kinase activity), and Y increases from bottom to top along the sigmoidal curve as the concentration increases.

[0273] Table 1 IC 50 (nM)

[0274]

[0275]

[0276] From the results in Table 1, it can be seen that the pyrimidinamine compounds of this application have excellent inhibitory activity against both NUAK1 / NUAK2, and are a dual-target small molecule kinase inhibitor. The IC 50 of the compound against NUAK1 can be lower than 1 nM, and the IC 50 against NUAK2 can reach several nM or dozens of nM. Therefore, through the above experiments, it has been proven that the pyrimidinamine compounds of this application can be used as NUAK1 / NUAK2 inhibitors.

[0277] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0278] In addition, it should be noted that, in the various specific technical features described in the above specific embodiments, they can be combined in any appropriate manner without contradiction. To avoid unnecessary repetition, the present invention will not separately describe various possible combination manners.

[0279] Furthermore, any combination can be made between various different embodiments of the present invention, as long as it does not violate the idea of the present invention, and it should also be regarded as the content disclosed by the present invention.

Claims

1. A pyrimidineamine compound, characterized in that, The pyrimidineamine compound is a compound represented by formula I-1 or I-2, or a stereoisomer, tautomer, isotope derivative, and pharmaceutically acceptable salt thereof; wherein X is a direct bond, -CR8R9-, and Y is -CO- or -S(O)2-; A is one of the following optionally substituted groups: The substituent on A is selected from one or more of halogen, amino, hydroxyl, 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, R8, and R9 are each independently H or an optionally substituted C1-8 alkyl group; R3 and R4 are each independently a halogen, amino group, hydroxyl group, nitro group, cyano group, mercapto group, optionally substituted C1-8 alkyl group, optionally substituted C1-8 alkoxy group, optionally substituted C1-8 alkylthio group, or optionally substituted C1-8 alkylamino group; R5 is an optionally substituted 3-6 membered cycloalkyl group, and the substituents on the cycloalkyl group are selected from one or more of a halogen, amino group, hydroxyl group, nitro group, cyano group, mercapto group, optionally substituted C1-8 alkyl group, optionally substituted C1-8 alkoxy group, optionally substituted C1-8 alkylthio group, and optionally substituted C1-8 alkylamino group; R 10 selected from the group consisting of halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino; R 11 is H or an optionally substituted C1-8 alkyl group; The substituents on the C1-8 alkyl group are selected from one or more of a halogen, amino group, hydroxyl group, nitro group, cyano group, and mercapto group; n is 0, 1, or 2, m is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, or 4.

2. The compound according to claim 1, wherein R5 is a halogen-substituted 3-6 membered cycloalkyl group.

3. The compound according to claim 2, wherein R5 is a fluorine-substituted 3-6 membered cycloalkyl group.

4. The compound according to claim 1, characterized in that, R5 is 5. The compound according to claim 4, characterized in that, R5 is 6. The compound according to any one of claims 1-5, characterized in that, R6 is methyl.

7. The compound according to any one of claims 1-5, characterized in that, R2 is H.

8. The compound according to any one of claims 1-5, characterized in that, R 11 is H or methyl.

9. The compound according to any one of claims 1-5, characterized in that, R3 is chlorine.

10. The compound according to claim 9, characterized in that, R3 is substituted at the para position of the amino group.

11. The compound according to any one of claims 1-5, characterized in that, The substituent on the A ring is a methoxy group.

12. The compound according to claim 11, wherein, The substituent on the A ring is substituted at the ortho position of the amino group.

13. The compound according to claim 1, characterized in that, The compound represented by formula I-1 or formula I-2 is the following compound:

14. The compound according to any one of claims 1-5, 13, wherein the pharmaceutically acceptable salt is formate.

15. The compound according to any one of claims 1-5, 13, wherein the isotope derivative is a deuterated compound.

16. A method for preparing the compound according to any one of claims 1-15, comprising the following steps: Preparing a compound of formula I-1 or formula I-2 from a compound of formula II-1 or formula II-2 and a compound of formula III; or Preparing a compound of formula V-1 or formula V-2 from a compound of formula IV and a compound of formula II-1 or formula II-2, and then preparing a compound of formula I-1 or formula I-2 from the compound of formula V-1 or formula V-2; Optionally, necessary protection and deprotection steps are included during the preparation process; Among them, Z is a leaving group, and the definitions of the other groups are the same as those defined in claims 1-15.

17. The preparation method according to claim 16, wherein The Z is a halogen.

18. The preparation method according to claim 17, wherein, The Z is Cl.

19. A pharmaceutical composition, characterized in that Using the compound according to any one of claims 1-15 as an active ingredient.

20. The pharmaceutical composition according to claim 19, characterized in that Containing a pharmaceutically acceptable carrier or excipient.

21. Use of the compound according to any one of claims 1-15 for preparing an inhibitor of NUAK1 or NUAK2. Use of the compound according to any one of claims 1 - 15 for the preparation of 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.

23. The use according to claim 22, characterized in that, The disease is Parkinson's disease, Alzheimer's disease, diabetes, hyperlipidemia, obesity, liver cancer, leukemia, lymphoma, tumor metastasis, liver cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or is used alone to reduce scars after trauma and surgery.

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