A pyrimidineamine NUAK inhibitor and its preparation method and use

Abstract

The present application provides a pyrimidineamine compound, characterized in that the pyrimidineamine compound is a compound represented by Formula I, or its stereoisomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs, and pharmaceutically acceptable salts. The pyrimidineamine compound described herein has excellent NUAK1 / NUAK2 kinase inhibitory activity and can be used to prevent or treat the following diseases by inhibiting NUAK1 / NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis, and skin fibrosis. #imgabs0#

Description

Technical Field

[0001] 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

[0002] 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 a phosphate group 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).

[0003] 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).

[0004] 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).

[0005] 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).

[0006] 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).

[0007] 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).

[0008] 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. Heterozygous NUAK1 deficiency 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 deficiency 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).

[0009] 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.

[0010] 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.

[0011] 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. Summary of the Invention

[0012] 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).

[0013] 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:

[0014]

[0015] wherein A is a phenyl group which may be substituted by m R3s, or a 5-membered heteroaryl group which may be substituted by p R4s;

[0016] B is an optionally substituted 3-10 membered cycloalkyl group or an optionally substituted 3-10 membered heterocycloalkyl group;

[0017] 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;

[0018] D is X is N or CH. When X is CH, H may be replaced by R5 or R6;

[0019] E is -C(O)-NHR7R8, -C(O)-R7R8, -NR7-C(O)R8, -NR7-S(O)2-R8, -optionally substituted C3-8heterocycloalkyl-S(O)2-R8, -optionally substituted C3-8cycloalkyl-S(O)2-R8, R8 is optionally substituted C1-8 alkyl, optionally substituted C3-8cycloalkyl, optionally substituted C3-8cycloalkylC1-8 alkyl, and the substituents on the cycloalkyl or heterocycloalkyl are 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;

[0020] R1 and R7 are each independently H, optionally substituted C1-8 alkyl;

[0021] R2, R3, R4, R5, and R6 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;

[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, m is 0, 1, 2, 3 or 4, and p, q, r are 0, 1, 2 or 3 respectively.

[0024] In one group of embodiments, E is one of the following formulae:

[0025]

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

[0027] Furthermore, B is one of the following formulae:

[0028]

[0029] R9 is 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;

[0030] R 10is H or optionally substituted C1-8 alkyl;

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

[0032] s and t are 0, 1, 2, 3 or 4 respectively, and u is 0, 1, 2 or 3.

[0033] Furthermore, B is one of the following formulae:

[0034]

[0035] In one group of embodiments, A is one of the following optionally substituted groups:

[0036]

[0037] The top of A is connected to the amino group, and the bottom of A is connected to the B group.

[0038] Furthermore, A is an optionally substituted phenyl group; furthermore, A is substituted by a methoxy group, preferably substituted at the ortho position of the amino group.

[0039] In one group of embodiments, R2 is chloro, preferably substituted in the para position relative to the amino group.

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

[0041]

[0042]

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

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

[0045] On the other hand, the present application also provides a method for preparing the pyrimidineamine compound, which comprises the following steps:

[0046] Prepare a compound of formula I from a compound of formula II and a compound of formula III;

[0047] or

[0048] When D is When the compound of formula V is prepared from the compound of formula IV and formula II, the compound of formula I is then prepared from the compound of formula V;

[0049]

[0050] Wherein, Y is a leaving group, preferably Y is halogen, more preferably Y is Cl, and the definitions of the other groups are as described above.

[0051] 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.

[0052] In one set of embodiments, when D is When the imino group on the D ring of the compound of formula IV is protected, the amino group or imino group on the B ring of formula II is optionally protected, and then the protected compound of formula IV is coupled with the optionally protected compound of formula II, and the substituent on the D ring is removed to prepare the compound of formula V, and finally the coupling reaction is carried out and the protecting group on the B ring is optionally removed to obtain the compound of formula I.

[0053] 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.

[0054] The present application provides the use of the pyrimidineamine compounds for preparing NUAK1 or NUAK2 inhibitors.

[0055] 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.

[0056] In one embodiment, 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.

[0057] The beneficial effects of the present invention are:

[0058] 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

[0059] 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.

[0060] 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.

[0061] 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.

[0062] definition

[0063] Unless otherwise specified, the following terms used in the present invention have the following definitions.

[0064] 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.

[0065] In the present invention, the ring groups connected to It represents that there may be n R2, q R5, r R6, s R9, t R9, or u R9 connected to any possible position of the ring group.

[0066] In the present invention, some substituents The place represents the connection site. When , the left side of D is connected to pyrimidine, and the right side of D is connected to the E group.

[0067] 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.

[0068] 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-membered aromatic monocyclic ring containing 1-2 atoms selected from N or S, or a 5-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.

[0069] 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.

[0070] 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.

[0071] 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.

[0072] 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.

[0073] The term "halogen" refers to F, Cl, Br, I.

[0074] 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.

[0075] 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 physical and chemical 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.

[0076] 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.

[0077] 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, 3H, 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.

[0078] The term "hydrate" refers to an association of one or more water molecules with a compound of the present invention.

[0079] The term "solvate" refers to an association between one or more solvent molecules and a compound of the present invention.

[0080] 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.

[0081] 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.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] Hereinafter, the effects of the specific compounds of the present invention will be described in detail with reference to Examples.

[0086] Example

[0087] Example 1 General method for synthesizing compound 534 (TDM-181134)

[0088]

[0089] Step 1: Compound 534c

[0090] tert-Butyl 4-(2-chloropyrimidin-4-yl)-1H-pyrazole-1-carboxylate

[0091] To a 100 mL three-necked flask were added compound 534a (1 g, 6.7 mmol), compound 534b (1.97 g, 6.7 mmol), 1,1'-bis(diphenylphosphinoferrocenepalladium)dichloride (490 mg, 0.67 mmol), cesium carbonate (4.4 g, 2 mmol), 1,4-dioxane (60 mL) and water (6 mL). The reaction solution was replaced with argon several times, heated to 70°C and stirred for one hour. The reaction was detected to be complete. Post-treatment: the reaction solution was poured into ice water (200 mL), and the aqueous phase was extracted twice with ethyl acetate (3*100 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography [eluent: (PE / EA) = 0-30%] to give the title compound (compound 534c, 900 mg, 47.8% yield) as a yellow solid. LCMS [M+1] + =282.

[0092] Step 2: Compound 534d

[0093] 2-Chloro-4-(1H-pyrazol-4-yl)pyrimidine

[0094] To a solution of compound 534c (1 g, 3.56 mmol) in dichloromethane (80 mL) and methanol (8 mL) was added a solution of hydrochloric acid-dioxane (8.9 mL, 35.6 mmol). The reaction mixture was heated to 40°C and stirred for 6 hours. The reaction was detected to be complete. Post-treatment: the reaction mixture was directly drained to obtain the target compound (compound 534d, 850 mg, yield 68%) as a white solid. LCMS [M+1] + =181.

[0095] Step 3: Compound 534f

[0096] tert-Butyl 3-(4-(2-chloropyrimidin-4-yl)-1H-pyrazol-1-yl)pyrrolidine-1-carboxylate

[0097] To a solution of compound 534d (755.2 mg, 4.18 mmol) in acetonitrile (150 mL) were added compound 534e (1.255 g, 5.02 mmol) and cesium carbonate (4.08 g, 12.54 mmol). The reaction mixture was heated to 50°C and stirred overnight. The reaction was detected to be complete. Post-treatment: The reaction mixture was directly drained, extracted with water and ethyl acetate, and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and drained. The crude product was purified by column chromatography (eluent: EA / PE = 0-50%) to obtain the title compound (compound 534f, 136 mg, 9.3% yield) as a white solid. LCMS [M+1] + =350.

[0098] Step 4: Compound 534g

[0099] 2-Chloro-4-(1-(pyrrolidin-3-yl)-1H-pyrazol-4-yl)pyrimidine

[0100] To a solution of compound 534f (136 mg, 0.39 mmol) in dichloromethane (15 mL) was added a solution of hydrochloric acid-dioxane (1.5 mL, 0.86 mmol). The reaction mixture was stirred at room temperature for 2 hours, and the reaction was determined to be complete. Post-treatment: The reaction mixture was directly drained to obtain the title compound (compound 534g, 97 mg, 93% yield) as a white solid. LCMS [M+1] + =250.

[0101] Step 5: Compound 534i

[0102] 2-Chloro-4-(1-(ethylsulfonyl)pyrrolidin-3-yl)-1H-pyrazol-4-yl)pyrimidine

[0103] To a solution of compound 534g (97.38 mg, 0.39 mmol) in N,N-dimethylformamide (6 mL) were added triethylamine (78.9 mg, 0.78 mmol) and compound 534h (75.2 mg, 0.59 mmol). The reaction mixture was stirred at room temperature for 2 hours, and the reaction was determined to be complete. Post-treatment: The reaction mixture was poured into water, and the aqueous phase was extracted with ethyl acetate (3 x 50 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography (eluent: EA / PE = 0-50%) to afford the title compound (compound 534i, 70 mg, yield 52.5%) as a white solid. LCMS [M+1] + =342.

[0104] Step 6: Compound 534k

[0105] tert-Butyl-4-(4-((4-(1-(ethylsulfonyl)pyrrolidin-3-yl)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0106] To a 100 mL three-necked flask were added compound 534i (48.5 mg, 0.14 mmol), compound 534j (45.3 mg, 0.17 mmol), palladium acetate (6.37 mg, 0.03 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (32.84 mg, 0.06 mmol), cesium carbonate (92.46 mg, 0.28 mmol), and 1,4-dioxane (8 mL). The reaction solution was replaced with argon several times, heated to 100 °C and stirred for 2 hours. The reaction was detected to be complete. Post-treatment: The reaction solution was drained, ethyl acetate (100 mL) and water (100 mL) were added for extraction, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and drained. The crude product was purified by column chromatography [eluent: (dichloromethane:methanol=10:1) / DCM=0-50%] to give the title compound as a white solid (compound 534k, 59 mg, yield 50.5%), LCMS [M+1] + =573.

[0107] Step 7: Compound 534

[0108] 4-(1-(1-(ethylsulfonyl)pyrrolidin-3-yl)-1H-pyrazol-4-yl)-N-(1-(piperidin-4-yl]-1H-pyridin-4-yl]pyrimidin-2-amine

[0109] To a solution of compound 534k (59.1 mg, 0.1 mmol) in dichloromethane (5 mL) and methanol (1 mL) was added a solution of hydrochloric acid-dioxane (0.52 mL, 2.07 mmol). The reaction was stirred at room temperature for two hours, and the reaction was determined to be complete. Post-treatment: The reaction solution was drained and the crude product was purified by preparative purification to obtain the title compound (compound 534, 7.7 mg, 5.5% yield) as a white solid. LCMS [M+1] + =472.3.

[0110] 1H NMR (400MHz, DMSO) δ9.37 (s, 1H), 8.53 (s, 1H), 8.35 (d, J = 5.0Hz, 2H), 8.16 (s, 1H), 7.97 (s ,1H),7.57(s,1H),7.00(d,J=5.1Hz,1H),5.20-5.09(m,1H),4.31(s,1H),3.84-3.79(m,2H ),3.63(d,J=6.3Hz,1H),3.53(d,J=5.6Hz,2H),3.21-3.11(m,4H),2.77(t,J=11.9Hz,2H) ,2.47-2.38(m,2H),2.03(d,J=11.2Hz,2H),1.92(d,J=11.1Hz,2H),1.22(t,J=7.4Hz,3H).

[0111] Example 2 General method for the synthesis of compound 535 (TDM-181135)

[0112]

[0113] Step 1: Compound 535c

[0114] 2-Chloro-4-nitropyrazolopyrimidine

[0115] Compound 535a (2 g, 0.013 mol) and compound 535b (1.44 g, 0.013 mol) in acetonitrile (24 mL) were added compound N, N-diisopropylethylamine (1.91 g, 0.01 mol), and the reaction solution was stirred at room temperature for 16 hours. After the reaction, the mixture was concentrated under reduced pressure, water (60 mL) was added to the residue and extracted with ethyl acetate (3*30 mL), the organic layers were combined, washed with saturated brine, 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 to 24 / 76) to obtain a white solid compound (compound 535c, 713.3 mg, 23.6% yield). LCMS [M+1] + =226.0.

[0116] Step 2: Compound 535d

[0117] 2-Chloropyrimidin-4-ylpyrazolamine

[0118] To a solution of compound 535c (50 mg, 0.222 mmol), ammonium chloride (59.27 mg, 1.11 mmol) and iron powder (61.89 mg, 1.108 mmol) in tetrahydrofuran (7.4 mL) was added water (7.4 mL), and the reaction solution was stirred at 65°C for 16 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, water (60 mL) was added to the residue and extracted with ethyl acetate (3*30 mL). The organic layers were combined, washed with saturated brine, and dried over sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-38 / 62) to obtain a yellow solid product (compound 535d, 45 mg, crude product). LCMS [M+1] + =196.0.

[0119] Step 3: Compound 535f

[0120] 1-(2-chloropyrimidin-4-yl)-4-pyrazolylpropanesulfonamide

[0121] To a dichloromethane solution of compound 535d (43.36 mg, 0.222 mmol) were added triethylamine (67.29 mg, 0.67 mmol) and compound 535e (63.17 mg, 0.443 mmol), and the mixture was stirred at 40°C for 16 hours. After the reaction, the mixture was concentrated under reduced pressure, water (30 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 (dichloromethane: dichloromethane containing 10% methanol = 0 to 44 / 56) to give a white solid compound (compound 535f, 42.9 mg, yield 64%), LCMS [M+1]. + =302.1.

[0122] Step 4: Compound 535h

[0123] tert-Butyl 4-(4-(4-propylsulfonamido)-1H-pyrazol-1-ylpyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0124] To a solution of compound 535f (50 mg, 0.166 mmol) and compound 535g (53.05 mg, 0.2 mmol) in dioxane hydrochloride (3 mL) were added 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (40.5 mg, 0.07 mmol), palladium acetate (6.74 mg, 0.03 mmol), and cesium carbonate (107.52 mg, 0.33 mmol). The mixture was degassed under vacuum, replaced with Ar, 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 yellow solid product (compound 535h, 64.9 mg, 73.5% yield). LCMS [M+1] + =532.2.

[0125] Step 5: Compound 535

[0126] 2-(1-(Piperidin-4-yl)-1H-pyrazol-4-yl)aminopyrimidin-4-yl)-1H-pyrazol-4-ylpropanesulfonamide

[0127] To a solution A (dichloromethane:methanol = 10:1) of compound 535h (64.90 mg, 0.122 mmol) was added dioxane hydrochloride (0.61 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 afford a white solid compound (compound 535, 19.8 mg, 37.6% yield). LCMS [M+1] + =532.2.

[0128] 1 H NMR(400,DMSO)δ9.73(s,1H),8.48(d,J=4.6Hz,1H),8.35(d,J=5.8Hz,2H), 7.95(s,1H),7.73(s,1H),7.56(s,1H),7.12(d,J=5.4Hz,1H),4.36-4.27(m ,2H),3.22(d,J=12.6Hz,3H),3.11-3.06(m,2H),2.82(t,J=11.8Hz,3H),2. 06(s,2H),1.98(s,2H),1.70(dd,J=15.1,7.5Hz,2H),0.96(t,J=7.4Hz,4H).

[0129] 2-(2-methoxy-4-(4-methylpiperazine-1-phenyl)aminopyrimidin-4-yl)-1H-pyrazol-4-ylpropanesulfonamide (TDM-181136) was synthesized as a yellow solid (16.9 mg, 21% yield) using a similar method to Example 2; and 2-(2-(2-methoxy-4-piperidinylphenyl)aminopyrimidin-4-yl)-4-ylpyrazolylpropane-1-sulfonamide (TDM-181138) was synthesized as a white solid (7.5 mg, 16.3% yield).

[0130]

[0131]

[0132] Example 3 General method for the synthesis of compound 539 (TDM-181139)

[0133]

[0134] Step 1: Compound 539c

[0135] N-(Cyanomethyl)-1H-pyrazole-4-carboxamide

[0136] To a solution of compound 539b (124 mg, 1.338 mmol) in acetonitrile (5 mL) was added N,N-diisopropylethylamine (472 mg, 3.65 mmol), and the mixture was stirred. 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (407 mg, 1.07 mmol) and compound 539a (100 mg, 0.892 mmol) were then added, and 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 (10% methanol (NH3) in dichloromethane / dichloromethane = 0 to 80 / 20) to give a yellow oily product (compound 539c, 402 mg, crude product). LCMS [M+1] + =151.

[0137] Step 2: Compound 539e

[0138] 1-(2-chloropyrimidin-4-yl)-N-(cyanomethyl)-1H-pyrazole-4-carboxamide

[0139] Potassium carbonate (370 mg, 2.676 mmol) was added to a solution of compound 539d (133 mg, 0.892 mmol) and compound 539c (crude product) in acetonitrile (10 mL), and the mixture was heated to 50 degrees and stirred overnight. The mixture was added to water (100 mL) and extracted with ethyl acetate (40 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 = 0-40 / 60) to give a white solid product (compound 539e, 91 mg, 38.8% yield). LCMS [M+1] + =249.

[0140] 1 H NMR (400MHz, DMSO) δ9.28–9.22(m,2H),8.89(d,J=5.5Hz,1H),8.34(s,1H),7.98(d,J=5.5Hz,1H),4.34(d,J=5.5Hz,2H).

[0141] Step 3: Compound 539

[0142] N-(Cyanomethyl)-1-(2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-pyrazole-4-carboxamide

[0143] To a mixture of compound 539e (45 mg, 0.171 mmol), compound 539f (45.4 mg, 0.206 mmol), palladium acetate (7.7 mg, 0.034 mmol), xantphos (40 mg, 0.068 mmol) and cesium carbonate (111 mg, 0.342 mmol) was added dioxane (5 mL). The mixture was degassed under vacuum, replaced with argon several times, heated to 100 degrees 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 = 0-40 / 60) and preparative HPLC (mobile phase containing 0.5 parts per thousand formic acid) to give a brown solid product (compound 539, TDM-181139, 7.6 mg, 9% yield). LCMS [M+1] + =448.

[0144] 1H NMR (400MHz, DMSO) δ9.25(t,J=5.4Hz,1H),8.99(s,1H),8.50(d,J=5.3Hz,1H),8.35(s,1H),8.26(s,1H),8.15(s,1H),7.66(t,J=8.9Hz,1H),7.19 (d,J=5.3Hz,1H),6.66(d,J=2.3Hz,1H),6.54(dd,J=8.7,2.3Hz,1H),4.3 7-4.28(m,2H),3.81(s,3H),3.20-3.14(m,4H),2.53(s,4H),2.27(s,3H).

[0145] Example 4 General method for the synthesis of compound 540 (TDM-181140)

[0146]

[0147] Step 1: Compound 540c

[0148] tert-Butyl-4-(4-((4-(4-(cyanomethyl)carbamoyl)-1H-pyrazol-1-yl)pyrimidin-2-yl)amino)-1H-pyridin-1-yl)piperidine-1-carboxylate

[0149] To a mixture of compound 540a (100 mg, 0.381 mmol), compound 540b (122 mg, 0.457 mmol), palladium acetate (17 mg, 0.076 mmol), xantphos (88 mg, 0.152 mmol) and cesium carbonate (248 mg, 0.762 mmol) was added dioxane (8 mL). The mixture was degassed under vacuum, replaced with argon several times, heated to 100 degrees 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=0-45 / 55) to give a yellow solid product (compound 540c, 146 mg, 77.8% yield). LCMS [M+1-100] + =393.

[0150] Step 2: Compound 540

[0151] N-(Cyanomethyl)-1-(2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)1H-pyrazole-4-carboxamide

[0152] To a solution of compound 540c (60 mg, 0.122 mmol) in dichloromethane (5 mL) and methanol (1 mL) was added zinc bromide (220 mg, 0.976 mmol) under argon atmosphere, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to give a yellow solid product (compound 540, TDM-181140, 17.4 mg, 36.3% yield). LCMS [M+1] + =393.

[0153] 1 H NMR(400MHz,MeOD)δ9.09(s,1H),8.50(d,J=5.3Hz,1H),8.17(s,1H),8.12(s,1H),7.67(s,1H),7.28(d ,J=5.4Hz,1H),4.60(s,1H),4.34(s,2H),3.58(d,J=13.0Hz,2H),3.28-3.17(m,2H),2.41-2.20(m,4H).

[0154] Example 5 General method for the synthesis of compound 545 (TDM-181145)

[0155]

[0156] Step 1: Compound 545c

[0157] tert-Butyl 3-(3-methoxy-4-nitrophenyl)-2,5-dihydropyrrole-1-carboxylate

[0158] To a mixture of compound 545a (534.6 mg, 2.85 mmol), compound 545b (842.0 mg, 2.85 mmol), 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride (104.63 mg, 0.14 mmol) and sodium carbonate (694.76 mg, 6.56 mmol) were added dioxane (20 mL) and water (5 mL), and then the mixture was degassed under vacuum and replaced with nitrogen. The mixture was heated to 102°C and stirred for 16 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, water (100 mL) was added to the residue, and the mixture was extracted with ethyl acetate (60*3). The organic layers were combined, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0-44 / 56). After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 0-76 / 24) to obtain a yellow solid compound (Compound 545c, 478.3 mg, 52.4% yield). LCMS [M+1-56] + =265.1.

[0159] Step 2: Compound 545d

[0160] 3-Methoxy-4-nitrophenyl-2,5-dihydropyrrole

[0161] To a solution A (dichloromethane: methanol = 10: 1) (15 mL) of compound 545c (378.3 mg, 1.181 mmol) was added dioxane hydrochloride (5.91 mL), and the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, water (60 mL) was added to the residue, and extracted with ethyl acetate (30 * 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 reaction was completed and the mixture was concentrated under reduced pressure to obtain a brown solid compound (compound 545d, 194.6 mg, 74.8% yield). LCMS [M+1] + =221.2.

[0162] Step 3: Compound 545e

[0163] 3-Methoxy-4-nitrophenyl-2,5-dihydropyrrole

[0164] To a solution of compound 545d (194.6 mg, 0.884 mmol) in methanol (12 mL) was added formaldehyde (214.86 mg). The mixture was stirred at room temperature for half an hour, followed by the addition of sodium acetate borohydride (749.1 mg, 3.53 mmol). The reaction mixture was stirred for another ten minutes. Upon completion of the reaction, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 0% to 100%) to afford a yellow solid compound (compound 545e, 85.3 mg, 41.2% yield). LCMS [M+1] + =235.2.

[0165] Step 4: Compound 545f

[0166] 2-Methoxy-4-(1-methylpyrrolidin-3-yl)aniline

[0167] To a solution of compound 545e (85.3 mg, 0.36 mmol) in methanol (3 mL) was added palladium on carbon (20 mg) at room temperature, and the mixture was then degassed under vacuum, replaced with hydrogen, and stirred for 3 hours. Upon completion of the reaction, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane:dichloromethane containing 10% methanol = 0-77 / 23) to give a yellow solid compound (compound 545f, 29 mg, 38.7% yield). Upon completion of the reaction, the mixture was concentrated under reduced pressure, and the residue was used directly in the next reaction. LCMS [M+1] +=207.2.

[0168] Step 5: Compound 545

[0169] N-cyanomethyl-1-(2-methoxy-4-(1-methylpyrrolidin-3-phenyl)amino)pyrimidin-4-yl)-1H-pyrazole-4-carboxamide

[0170] To a solution of compound 545f (36.76 mg, 0.14 mmol) and compound 545g (29 mg, 0.14 mmol) in dioxane (4 mL) were added 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (32.4 mg, 0.06 mmol), palladium acetate (6.27 mg, 0.028 mmol), and cesium carbonate (91.23 mg, 0.28 mmol). The mixture was degassed under vacuum, replaced with argon, and the reaction solution was stirred at 100° C. for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, water (30 mL) was added to the residue, and the mixture was extracted with ethyl acetate (30*3). The organic layers were combined, washed with brine (50 mL), and dried over sodium sulfate. The filtrate was concentrated under reduced pressure. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane:dichloromethane containing 10% methanol = 0-100 / 0) and then purified by preparative HPLC (formic acid) to obtain a white solid compound (Compound 545, 6.5 mg, 10.7% yield). LCMS [M+1] + =433.2.

[0171] 1 H NMR (400MHz, DMSO) δ9.28(s,1H),9.06(s,1H),8.57(d,J=5.3Hz,1H),8.39(s,1H),8.26(d,J=11 .4Hz,2H),7.94(d,J=8.1Hz,1H),7.27(d,J=5.3Hz,1H),7.01(s,1H),6.94(d,J=8.1Hz,1H),4.34 (d,J=5.2Hz,2H),3.86(s,3H),3.42-3.33(m,1H),3.04(t,J=8.6Hz,1H),2.80(t,J=6.9Hz,2H),2 .64(t,J=8.5Hz,1H),2.43(s,3H),2.36-2.22(m,1H),1.87(dd,J=12.6,7.6Hz,1H),1.23(s,1H).

[0172] Example 6 General method for the synthesis of compound 546 (TDM-181146)

[0173]

[0174] Step 1: Compound 546c

[0175] 1-(2-chloropyrimidin-4-yl)-1H-pyrrole-3-carboxylic acid methyl ester

[0176] To a solution of compound 546b (1 g, 9.59 mmol) in N,N-dimethylformamide (20 mL) at 0°C was added sodium hydroxide (384 mg, 9.59 mmol). The reaction mixture was warmed to room temperature and stirred for 30 minutes. The temperature was then lowered to 0°C, and compound 546a (1.43 g, 9.59 mmol) was added to the solution. The reaction mixture was stirred at room temperature for 3 hours. The reaction was determined to be complete. Post-treatment: the reaction mixture was poured into water and extracted with ethyl acetate (60 mL x 3). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography (eluent: (EA:PE) = 0-15%) to afford the title compound (compound 546c, 987 mg, yield 41%) as a white solid. LCMS [M+1] + =238.

[0177] 1 H NMR (400MHz, DMSO) δ8.83(d,J=5.7Hz,1H),8.39(s,1H),8.08(d,J=5.7Hz,1H),7.88-7.79(m,1H),6.74(dd,J=3.1,1.4Hz,1H),3.79(s,3H).

[0178] Step 2: Compound 546d

[0179] 1-(2-Chloropyrimidin-4-yl)-1H-pyrrole-3-carboxylic acid

[0180] A solution of compound 546c (550 mg, 2.3 mmol) in acetic acid (17.5 mL) and concentrated hydrochloric acid (8.75 mL) was heated to 50°C and stirred for 7 hours. The reaction was detected to be complete. Post-treatment: the reaction solution was poured into water, and the aqueous phase was extracted with ethyl acetate (2*100 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography [eluent: (dichloromethane:methanol=10:1) / dichloromethane=0-80%] to give the title compound (compound 546d, 95.8 mg, yield 18.6%) as a white solid. LCMS [M+1] + =224.

[0181] Step 3: Compound 546f

[0182] 1-(2-chloropyrimidin-4-yl)-N-(cyanomethyl)-1H-pyrrole-3-carboxamide

[0183] To a solution of compound 546d (64 mg, 0.29 mmol) in N,N-dimethylformamide (10 mL) were added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (163.1 mg, 0.43 mmol), N,N-diisopropylethylamine (92.4 mg, 0.72 mmol) and compound 546e (26.4 mg, 0.29 mmol). The reaction solution was stirred at room temperature for 2 hours and the reaction was detected to be complete. Post-treatment: The reaction solution was poured into water, extracted with ethyl acetate (3*100 mL), and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography [eluent: (dichloromethane:methanol=10:1) / dichloromethane=0-80%] to give the title compound as a white solid (compound 546f, 59.7 mg, yield 53.3%). LCMS [M+1] + =262.

[0184] Step 4: Compound 546

[0185] N-(Cyanomethyl)-1-(2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-pyrrole-3-carboxamide

[0186] To a 100 mL three-necked flask were added compound 546f (30 mg, 0.12 mmol), compound 546g (30.5 mg, 0.14 mmol), palladium acetate (5.16 mg, 0.02 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (26.6 mg, 0.05 mmol), cesium carbonate (75 mg, 0.23 mmol), and 1,4-dioxane (6 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 concentrated to dryness, then extracted with ethyl acetate (100 mL) and water (100 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and dried. The crude product was purified to yield the title compound (compound 546, 59 mg, 50.5% yield) as a white solid. LCMS [M+1] + =447.2.

[0187] 1H NMR (400MHz, DMSO) δ8.84(t,J=5.6Hz,1H),8.42(d,J=5.5Hz,1H),8.25(s,2H),8. 17(s,1H),7.72-7.67(m,1H),7.64(d,J=8.7Hz,1H),7.10(d,J=5.5Hz,1H),6.74(d d,J=3.2,1.6Hz,1H),6.65(d,J=2.4Hz,1H),6.53(dd,J=8.8,2.4Hz,1H),4.27(d, J=5.5Hz,2H),3.81(s,3H),3.19-3.12(m,4H),2.49(d,J=5.1Hz,4H),2.25(s,3H).

[0188] N-(Cyanomethyl)-1-(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-pyrrole-3-carboxamide (TDM-181147) was synthesized by a method similar to Example 6 as a white solid (11.8 mg, 24.6% yield).

[0189]

[0190]

[0191] Example 7 General method for the synthesis of compound 548 (TDM-181148)

[0192]

[0193] Step 1: Compound 548c

[0194] tert-Butyl-4-(4-((4-(4-(cyanomethyl)carbamoyl)-1H-pyrazol-1-yl)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidine-1-carboxylate

[0195] To a mixture of compound 548a (70 mg, 0.266 mmol), compound 548b (98 mg, 0.319 mmol), palladium acetate (12 mg, 0.053 mmol), xantphos (61.6 mg, 0.106 mmol) and cesium carbonate (173 mg, 0.532 mmol) was added dioxane (5 mL). The mixture was degassed under vacuum, replaced with argon several times, heated to 100 degrees 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 = 0-14 / 86) to give a yellow solid product (compound 548c, 81 mg, 57.2% yield). LCMS [M+1] + =533.

[0196] Step 2: Compound 548

[0197] N-(Cyanomethyl)-1-(2-((2-methoxy-4-(piperidin-4-yl)phenyl)amino)pyrimidin-4-yl)-1H-pyrazole-4-carboxamide

[0198] To a solution of compound 548c (73 mg, 0.137 mmol) in dichloromethane (5 mL) and methanol (0.5 mL) was added zinc bromide (247 mg, 1.096 mmol) under argon atmosphere, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to give the desired product (compound 548, TDM-181148, 6 mg, 10.1% yield) as a colorless oil. LCMS [M+1] + =433.

[0199] 1 H NMR (400MHz, DMSO) δ9.46 (s, 1H), 9.08 (s, 1H), 8.57 (d, J = 5.4Hz, 1H), 8.48 (s, 1H), 8. 40(s,1H),8.28(d,J=6.2Hz,1H),7.98(d,J=8.2Hz,1H),7.28(d,J=5.4Hz,1H),6.96( s,1H),6.88(d,J=8.2Hz,1H),4.34(d,J=4.9Hz,2H),3.87(s,3H),3.67(s,1H),3.32( d,J=11.6Hz,2H),2.92(t,J=11.5Hz,2H),2.80(t,J=11.5Hz,1H),1.98–1.76(m,4H).

[0200] Example 8: General method for the synthesis of compound 552 (TDM-181152)

[0201]

[0202] Step 1: Compound 552c

[0203] tert-Butyl 4-(4-((4-(3-((cyanomethyl)carbamoyl)-1H-pyrrol-1-yl)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidine-1-carboxylate

[0204] To a mixture of compound 552a (80 mg, 0.306 mmol), compound 552b (112 mg, 0.367 mmol), palladium acetate (14 mg, 0.061 mmol), xantphos (70.8 mg, 0.112 mmol) and cesium carbonate (199 mg, 0.612 mmol) was added dioxane (5 mL). The mixture was degassed under vacuum, replaced with argon several times, heated to 100 degrees 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=0-55 / 45) to give a yellow solid product (compound 552c, 96 mg, 59% yield). LCMS[M+1] + =532.

[0205] Step 2: Compound 552

[0206] N-(Cyanomethyl)-1-(2-((2-methoxy-4-(piperidin-4-yl)phenyl)amino)pyrimidin-4-yl)-1H-pyrrole-3-carboxamide

[0207] To a solution of compound 552c (96 mg, 0.181 mmol) in dichloromethane (10 mL) and methanol (1 mL) was added zinc bromide (325 mg, 1.445 mmol) under argon atmosphere, and the mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to give the product as a white solid (compound 552, TDM-181152, 20 mg, 25.6% yield). LCMS [M+1] + =432.

[0208] 1 H NMR (400MHz, DMSO) δ8.91(t,J=5.3Hz,1H),8.49(d,J=5.5Hz,1H),8.40(s,1H),8.31(d,J=6.7Hz ,2H),7.94(d,J=8.2Hz,1H),7.74(s,1H),7.19(d,J=5.5Hz,1H),6.93(s,1H),6.85(d,J=8.1Hz,1 H),6.77(s,1H),4.28(d,J=5.4Hz,2H),3.86(s,3H),3.66(s,1H),3.28(d,J=11.7Hz,2H),2.87( t,J=11.7Hz,2H),2.77(t,J=11.8Hz,1H),1.89(d,J=12.3Hz,2H),1.78(dd,J=22.6,12.1Hz,2H).

[0209] N-(Cyanomethyl)-1-(2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)1H-pyrrole-3-carboxamide (TDM-181153) was synthesized as an off-white solid (8.5 mg, yield, 9%) by a method similar to that of Example 8; and N-(Cyanomethyl)-1-(2-((4-(piperidin-4-yl)phenyl)amino)pyrimidin-4-yl]-1H-pyrrole-3-carboxamide (TDM-181154) was synthesized as a white solid (25.2 mg, yield 28.6%).

[0210]

[0211]

[0212] Example 9: General method for the synthesis of compound 555 (TDM-181155)

[0213]

[0214] 1-(2,5-dichloropyrimidin-4-yl)-1H-pyrrole-3-carboxylic acid methyl ester

[0215] Potassium carbonate (1131.9 mg, 8.19 mmol) was added to a solution of compound 555a (500 mg, 2.73 mmol) and compound 555b (341 mg, 2.73 mmol) in acetonitrile (50 mL). The reaction mixture was heated to 50°C and stirred overnight. The reaction was determined to be complete. Post-treatment: The reaction mixture was poured into water and extracted with ethyl acetate (2 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography (eluent: TLC, PE / EA = 3:1, PE / EA = 100:0-85:15) to afford the title compound (compound 555c, 510 mg, 68.72% yield) as a white solid. LCMS [M+1] + =272.

[0216] Step 2: Compound 555d

[0217] 1-(2,5-dichloropyrimidin-4-yl)-1H-pyrrole-3-carboxylic acid methyl ester

[0218] Compound 555c (1.1 g, 4.04 mmol) was added to a solution of acetic acid (30 mL) and concentrated hydrochloric acid (15 mL). The reaction mixture was heated to 50°C and stirred for 10 hours. The reaction was detected to be complete. Post-treatment: The reaction mixture was poured into water, and the aqueous phase was extracted with ethyl acetate (2 x 100 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography (eluent: TLC, dichloromethane / methanol = 10:1, dichloromethane / methanol = 100:0-20:80) to obtain the title compound (compound 555d, 315 mg, 30% yield) as a white solid. LCMS [M+1] + =258,260.

[0219] Step 3: Compound 555f

[0220] 1-(2-chloropyrimidin-4-yl)-N-(cyanomethyl)-1H-pyrrole-3-carboxamide

[0221] To a solution of compound 555d (170 mg, 0.66 mmol) in N,N-dimethylformamide (17 mL) were added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (375.7 mg, 0.99 mmol), N,N-diisopropylethylamine (212.5 mg, 1.65 mmol) and compound 555e (61.2 mg, 0.66 mmol). The reaction solution was stirred at room temperature for 10 minutes and the reaction was detected to be complete. Post-treatment: The reaction solution was poured into water, and the aqueous phase was extracted with ethyl acetate (3*100 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography [eluent: TLC, PE / EA=10:1, PE / EA=100:0-50:50 during column chromatography] to afford the title compound as a white solid (compound 555f, 117 mg, 54% yield). LCMS [M+1] + =296,298.

[0222] Step 4: Compound 555

[0223] 1-(5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-N-(cyanomethyl)-1H-pyrrole-3-carboxamide

[0224] To a 100 mL three-necked flask were added compound 555f (20 mg, 0.07 mmol), compound 555g (15 mg, 0.07 mmol), palladium acetate (3 mg, 0.01 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (15.6 mg, 0.03 mmol), cesium carbonate (44 mg, 0.14 mmol), and 1,4-dioxane (4 mL). The reaction mixture was purged with argon several times, heated to 100°C, and stirred for 2 hours. The reaction was complete. Post-treatment: The reaction mixture was concentrated to dryness, and the residue was extracted with ethyl acetate (100 mL) and water (100 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified to yield the title compound (compound 555, 3.2 mg, 9.8% yield) as a yellow solid. LCMS [M+1] + =481,483.

[0225] 1 H NMR(400MHz,DMSO)δ8.86(t,J=5.6Hz,1H),8.71(s,1H),8.53(s,1H),8.16( s,1H),8.14(s,1H),7.53(s,1H),7.41(d,J=8.7Hz,1H),6.73(dd,J=3.2,1. 6Hz,1H),6.63(d,J=2.4Hz,1H),6.50(dd,J=8.7,2.5Hz,1H),4.27(d,J=5.5 Hz, 2H), 3.78 (s, 3H), 3.17–3.12 (m, 4H), 2.47 (d, J = 4.9Hz, 4H), 2.24 (s, 3H).

[0226] N-(cyanomethyl)-1-(2-((2-methoxy-4-(1-methylpiperidin-4-yl)phenyl)amino)pyrimidin-4-yl-1H-pyrrole-3-carboxamide (TDM-181156) was synthesized by a method similar to that in Example 9 as a white solid (7.4 mg, yield, 16.8%); 1-(5-chloro-2-((2-methoxy-4-(piperidin-4-yl)phenyl)amino)pyrimidin-4-yl]-N-(cyanomethyl)-1H-pyrrole-3-carboxamide (TDM-181157) was synthesized as a white solid (27.7 mg, yield 47%); and N-cyanomethyl-1-(2-methoxy-4-methylpyrrolidin-3-phenyl)amino)pyrimidin-4-ylpyrrole-3-carboxamide (TDM-181158) was synthesized as a white solid (6.2 mg, 3.4% yield).

[0227]

[0228]

[0229] Example 10: General method for the synthesis of compound 559 (TDM-181159)

[0230]

[0231] Step 1: Compound 559c

[0232] 2-Chloro-4-nitropyrazolopyrimidine

[0233] Compound 559a (3 g, 0.02 mol) and compound 559b (430.74 mg, 2.01 mmol) in acetonitrile (36 mL) were added compound N, N-ethyldiisopropylamine (2.84 g, 0.03 mol), and the reaction solution was stirred at room temperature for 16 hours. After the reaction, the mixture was concentrated under reduced pressure, water (100 mL) was added to the residue, and 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, and the residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 0 to 76 / 24) to obtain a colorless solid compound (compound 559c, 1.334 g, 29.6% yield). LCMS [M+1] + =226.1.

[0234] Step 2: Compound 559e

[0235] tert-Butyl 4-(4-(4-nitropyrazol-1-yl)aminopyrimidin-1-amino)-1H-pyrazol-1-ylpiperidine-1-carboxylate

[0236] To a solution of compound 559c (50 mg, 0.23 mmol) and compound 559d (50.2 mg, 0.23 mmol) in dioxane (2 mL) were added 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (53.3 mg, 0.09 mmol), palladium acetate (10.3 mg, 0.046 mmol), and cesium carbonate (149.9 mg, 0.46 mmol). The mixture was degassed under vacuum, replaced with argon, and stirred at 100°C for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure, and water (30 mL) was added to the residue. The base was adjusted with potassium carbonate solution, and then extracted with ethyl acetate (3 x 20 mL). The organic layers were combined. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0-50 / 50) to give a yellow compound (compound 559e, 55.7 mg, 55.2% yield), LCMS [M+1-56] + =400.1.

[0237] Step 3: Compound 559f

[0238] tert-Butyl 4-(4-(4-(4-aminopyrazol-1-yl)aminopyrimidin-2-yl)amino-1H-pyrazol-1-ylpiperidine-1-carboxylate

[0239] To a solution of compound 559e (390.4 mg, 0.86 mmol) in tetrahydrofuran (25 mL) were added ammonium chloride (229.24 mg, 4.29 mmol), iron powder (239.35 mg, 4.29 mmol), and water (25 mL), and the mixture was stirred at 65°C for 6 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure. Water (60 mL) was added to the residue, and the mixture was extracted with ethyl acetate (40*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 (dichloromethane: dichloromethane containing 10% methanol = 0-43 / 57) to give a white solid compound (compound 559f, 203.7 mg, yield 55.7%), LCMS [M+1]. + =426.2.

[0240] Step 4: Compound 559h

[0241] 4-(4-(4-(4-(2,2-difluorocyclopropane-1-carboxamido)-1H-pyrazol-1-ylaminopyrimidine)-1H-pyrazol-1-ylaminopiperidine

[0242] To a solution of compound 559g (34.4 mg, 0.28 mmol) in N,N-dimethylformamide (5 mL) were added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (107.2 mg, 0.28 mmol) and N,N-diisopropylethylamine (36.4 mg, 0.28 mmol), and the mixture was stirred at room temperature for 10 minutes. Compound 559f (80 mg, 0.19 mmol) was then added, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was filtered and concentrated under reduced pressure. Water (30 mL) was added to the residue and extracted with ethyl acetate (20*5). The organic layers were combined, washed with brine (3*30 mL), and dried over sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography. 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 = 0-37 / 63) to obtain a yellow solid product (Compound 559h, 84.3 mg, 86.6% yield). LCMS [M+1] + =530.3.

[0243] Step 5: Compound 559

[0244] 2,2-Difluoro-N-(1-2-(1-piperidin-4-yl)-1H-pyrazol-4-amino)pyrimidin-4-yl)-1H-pyrazol-4-yl)cyclopropane-1-carboxamide

[0245] To a solution A (dichloromethane:methanol = 10:1) (3 mL) of compound 559f (36.76 mg, 0.14 mmol) was added dioxane hydrochloride (0.38 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 (TDM-181159, compound 559, 18.1 mg, 49.5% yield). LCMS [M+1] + =430.1.

[0246] 1 H NMR(400MHz,MeOD)δ8.94(s,1H),8.54(s,1H),8.40(d,J=5.5Hz,1H),8.20(s,1 H),7.76(s,1H),7.57(s,1H),7.22(d,J=5.5Hz,1H),4.51(t,J=11.1Hz,1H),3. 51(dd,J=7.8,3.6Hz,2H),3.21–3.12(m,2H),2.73–2.64(m,1H),2.38(s,2H),2 .24(dd,J=24.6,12.9Hz,2H),2.09(dt,J=13.7,6.9Hz,1H),1.96–1.86(m,1H).

[0247] Example 11: General method for the synthesis of compound 569 (TDM-181169)

[0248]

[0249] Step 1: Compound 569c

[0250] 1-Methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine

[0251] Under nitrogen, a 100 mL three-necked flask was charged with compound 569b (146.6 mg, 1.3 mmol), compound 569a (149.3 mg, 1.3 mmol), triphenylphosphine (682 mg, 2.6 mmol), and anhydrous tetrahydrofuran (10 mL). The temperature was then lowered to 0°C and diisopropyl azodicarboxylate (0.46 mL, 2.6 mmol) was added. The reaction mixture was stirred at room temperature overnight. The reaction was detected 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 and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography (eluent: (dichloromethane:methanol = 10:1) / DCM = 0-40%) to afford the title compound (compound 569c, 110 mg, 40.3% yield) as a yellow oil. LCMS [M+1] + =211.

[0252] Step 2: Compound 569d

[0253] 1-(1-Methylpiperidin-4-yl)-1H-pyrazol-4-amine

[0254] To a solution of compound 569c (120 mg, 0.57 mmol) in methanol (10 mL) was added palladium on carbon (10 mg). The reaction solution was purged several times with a hydrogen balloon and stirred at room temperature for two hours. The reaction was determined to be complete. Post-treatment: The reaction solution was filtered, the filter cake was washed with methanol, and the filtrate was concentrated to dryness to obtain the title compound (compound 569d, 83.7 mg, 81.5% yield) as a white solid. LCMS [M+H] + =181.

[0255] Step 3: Compound 569g

[0256] tert-Butyl 4-(2-chloropyrimidin-4-yl)-1H-pyrazole-1-carboxylate

[0257] To a 100 mL three-necked flask were added compound 569e (500 mg, 3.36 mmol), compound 569f (988 mg, 3.36 mol), 1,1'-bis(diphenylphosphinoferrocenepalladium) dichloride (254.80 mg, 0.34 mmol), cesium carbonate (2189.3 mg, 6.72 mmol), 1,4-dioxane (25 mL), and water (2.5 mL). The reaction mixture was purged with argon several times, heated to 70°C, and stirred for 1 hour. The reaction was then checked for completion. Post-treatment: The reaction solution was poured into water and extracted with ethyl acetate (3*400 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled dry. The crude product was passed through a column [eluent: (PE / EA) = 0-30%] to give the target compound as a white solid (compound 569 g, 588.1 mg, yield 62.4%). LCMS [M+H] + =281.

[0258] Step 4: Compound 569h

[0259] tert-Butyl-4-(2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)1H-pyrazole-1-carboxylate

[0260] To a 100 mL three-necked flask were added compound 569g (504.8 mg, 1.8 mmol), compound 569d (389 mg, 2.16 mmol), palladium acetate (80.8 mg, 0.36 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (416.6 mg, 0.72 mmol), cesium carbonate (1173 mg, 3.6 mmol), and 1,4-dioxane (25 mL). The reaction solution was replaced with argon several times, heated to 100 ° C and stirred for two hours. The reaction was detected to be complete. Post-treatment: The reaction solution was concentrated to dryness, ethyl acetate (100 mL) and water (100 mL) were added, and the mixture was separated and extracted. 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: (dichloromethane:methanol=10:1) / DCM=0-100%] to give the target compound (compound 569h, 225.8 mg, yield 29%) as a yellow solid. LCMS [M+H] + =425.

[0261] Step 5: Compound 569i

[0262] N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-4-(1H-pyrazol-4-)pyrimidin-2-amine

[0263] To a solution of compound 569h (225.8 mg, 0.53 mmol) in dichloromethane (20 mL) was added trifluoroacetic acid (2.2 mL). The reaction mixture was stirred at room temperature for one hour, and the reaction was detected to be complete. Post-treatment: The reaction mixture was poured into water and extracted twice with ethyl acetate (2 x 100 mL). The aqueous phases were combined and the pH was adjusted to alkaline. The aqueous phases were extracted twice with ethyl acetate (2 x 100 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled to dryness to obtain the title compound (compound 569i, 172.5 mg, 92% yield) as a white solid. LCMS [M+H] + =325.

[0264] Step 6: Compound 569

[0265] Cyclopropyl(4-(2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)1H-pyrazol-1-yl)methanone

[0266] To a solution of compound 569j (61.1 mg, 0.71 mmol) in N,N-dimethylformamide (5 mL) were added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (270 mg, 0.71 mmol), N,N-diisopropylethylamine (92 mg, 0.71 mmol), and compound 569i (152.5 mg, 0.47 mmol). The reaction mixture was heated to 50°C and stirred for 2 hours. The reaction was then determined to be complete. Post-treatment: The reaction solution was poured into water, and ethyl acetate (3*100 mL) was added and extracted three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled dry. The crude product was passed through a column chromatography [eluent: (dichloromethane:methanol=10:1) / DCM=0-50%] and recrystallized from methanol to obtain the target compound as a white solid (compound 569, 11.7 mg, yield 9.2%). LCMS [M+H] + =393.2.

[0267] 1 H NMR(400MHz,)δ9.56(s,1H),9.34(s,1H),9.12(s,1H),8.54(s,1H),8.46(d,J=5.1Hz,1H),8.04(s,1H),7.62(s,1H),7.28(d,J=5.1Hz ,1H),4.48(s,1H),3.57(d,J=12.1Hz,2H),3.12(ddd,J=12.3,7.8,4.6Hz,3H),2.85(s,3H),2.21(d,J=31.1Hz,4H),1.35-1.12(m,4H).

[0268] Example 12: General method for the synthesis of compound 570 (TDM-181170)

[0269]

[0270] Step 1: Compound 570c

[0271] 1-(2,5-dichloropyrimidin-4-yl)-1H-pyrrole-3-carboxylic acid methyl ester

[0272] Potassium carbonate (2.21 g, 16 mmol) was added to a solution of compound 570a (1.46 g, 8 mmol) and compound 570b (1 g, 8 mmol) in acetonitrile (100 mL). The reaction mixture was heated to 50°C and stirred overnight. The reaction was determined to be complete. Post-treatment: The reaction mixture was poured into water (150 mL) and extracted twice with ethyl acetate (2 x 200 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: TLC: PE / EA = 3:1, PE:EA = 100:0-85:15) to afford the title compound (compound 570c, 1.89 g, 87.5% yield) as a white solid. LCMS [M+1] + =272.

[0273] Step 2: Compound 570d

[0274] 1-(2,5-Dichloropyrimidin-4-yl)-1H-pyrrole-3-carboxylic acid

[0275] To a solution of compound 570c (1.89 g, 6.95 mmol) in acetic acid (40 mL) was added concentrated hydrochloric acid (20 mL). The reaction mixture was heated to 50°C and stirred for 15 hours. The reaction was determined to be complete. Post-treatment: The reaction mixture was poured into water and extracted twice with ethyl acetate (2 x 100 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 = 0-15%) to afford the title compound (compound 570d, 495 mg, 26.7% yield) as a white solid. LCMS [M+1] + =258,260.

[0276] Step 3: Compound 570f

[0277] 1-(2,5-Dichloropyrimidin-4-yl)-N-methyl-1H-pyrrole-3-carboxamide

[0278] To a solution of compound 570d (158.6 mg, 0.62 mmol) in N,N-dimethylformamide (15 mL) were added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (353.6 mg, 0.93 mmol), N,N-diisopropylethylamine (200 mg, 1.55 mmol), and compound 570e (41.8 mg, 0.62 mmol). The reaction was stirred at room temperature for 10 minutes, and the reaction was determined to be complete. Post-treatment: The reaction solution was poured into water, extracted three times with ethyl acetate (3*100 mL), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and pulled dry. The crude product was purified by column chromatography [eluent: (dichloromethane:methanol=10:1) / dichloromethane=0-15%] to give the target compound (compound 570f, 129.8 mg, yield 65%) as a yellow solid. LCMS [M+1] + =271,273.

[0279] Step 4: Compound 570

[0280] 1-(5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-N-methyl-1H-pyrrole-3-carboxamide

[0281] To a 100 mL three-necked flask were added compound 570f (104.8 mg, 0.39 mmol), compound 570g (88.7 mg, 0.46 mmol), palladium acetate (17.4 mg, 0.08 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (89.6 mg, 0.16 mmol), cesium carbonate (252.2 mg, 0.77 mmol), and 1,4-dioxane (20 mL). The reaction mixture was purged with argon several times, heated to 100°C, and stirred for two hours until the reaction was complete. Post-treatment: The reaction solution was concentrated to dryness, and the crude product was purified by column chromatography [eluent: (dichloromethane:methanol=10:1) / dichloromethane=0-50%]. The crude product was further purified by preparative purification to obtain the target compound as a yellow solid (Compound 570, 48.9 mg, 24% yield). LCMS [M+1] + =426.2.

[0282] 1H NMR (400MHz, DMSO-d6) δ9.79(s,1H),8.59(s,1H),8.16(s,1H),8.09(q,J=2.4,2.0Hz,2H),7.56(dd,J=3.3,2.3Hz,1H),7.54-7.46(m,2H ),6.98-6.86(m,2H),6.73(dd,J=3.3,1.7Hz,1H),3.08(dd,J=6.4,3.6Hz,4H),2.74(d,J=4.5Hz,3H),2.46(d,J=4.9Hz,4H),2.23(s,3H).

[0283] 1-(5-Chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-N-(cyclopropylmethyl)-1H-pyrrole-3-carboxamide (TDM-181171) was synthesized as a yellow solid (65 mg, yield, 26%) using a similar method to Example 12.

[0284]

[0285] Example 13: General method for the synthesis of compound 577 (TDM-181177)

[0286]

[0287] Step 1: Compound 577c

[0288] 2,5-Dichloro-4-(4-nitropyrazol-1-yl)pyrimidine

[0289] To a solution of compound 577a (1.02 g, 0.006 mol) in acetonitrile (20 mL) was added compound 577b (0.6 g, 0.0052 mol) and then N,N-diisopropylethylamine (0.78 g, 0.006 mol). The reaction mixture was heated to 30°C and stirred for 3 hours. LCMS [M+1] + =260, indicating the reaction was complete. Post-treatment: The reaction solution was concentrated to dryness, and the crude product was separated by adding ethyl acetate (120 mL) and water (60 mL). The separated organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The crude product was purified by column chromatography (eluent: EA / PE = 0-30%) to obtain the target compound (compound 577c, 1.02 g, yield 65.4%) as a white solid. LCMS [M+1] + =260.

[0290] Step 2: Compound 577d

[0291] 1-(2,5-Dichloropyrimidin-4-yl)-1H-pyrazol-4-amine

[0292] To a solution of compound 577c (917.80 mg, 3.53 mmol) in tetrahydrofuran (30 mL) were added ammonium chloride (944.10 mg, 17.65 mmol) and iron powder (985.64 mg, 17.65 mmol), followed by water (30 mL). The reaction mixture was heated to 65°C and stirred for 3 hours. LCMS [M+1] + =230.1, indicating the reaction was complete. Post-treatment: Filtration, the organic phase was separated by adding ethyl acetate (100 mL) and water (50 mL). The separated organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The crude product was purified by column chromatography (eluent: EA / PE = 0-50%) to obtain the target compound (compound 577d, 741 mg, 82% yield) as a yellow solid. LCMS [M+1] + =230.1.

[0293] Step 3: Compound 577f

[0294] N-(1-(2,5-dichloropyrimidin-4-yl)-1H-pyrazol-4-yl)propane-1-sulfonamide

[0295] To a solution of compound 577d (400 mg, 1.74 mmol) in anhydrous tetrahydrofuran (20 mL) at 0°C were added pyridine (275.07 mg, 3.48 mmol) and compound 577e (488.80 mg, 3.43 mmol). The reaction mixture was stirred at room temperature for one hour, replaced with nitrogen several times, and then the reaction mixture was heated to 40°C and stirred overnight. LCMS [M+1] + =336, indicating the reaction was complete. Post-treatment: The reaction solution was concentrated to dryness, and the crude product was separated by adding ethyl acetate (100 mL) and water (50 mL). The separated organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The crude product was purified by column chromatography (eluent: EA / PE = 0-40%) to give the target compound (Compound 577f, 531 mg, 86% yield) as a yellow liquid. LCMS [M+1] + =336.

[0296] Step 4: Compound 577h

[0297] tert-Butyl 4-((5-chloro-4-(4-propylsulfonamido)-1H-pyrazol-1-yl)pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0298] Compound 577f (100 mg, 0.297 mmol) was added with compound 577g (79.10 mg, 0.297 mmol), palladium acetate (13.34 mg, 0.059 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (68.74 mg, 0.119 mmol), and cesium carbonate (193.53 mg, 0.594 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-100] + =466.2, indicating the reaction was complete. Post-treatment: The reaction solution was added with water (30 mL) and ethyl acetate (60 mL), and the separated organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The crude product was purified by column chromatography (eluent: EA / PE = 0-50%) to give the target compound (compound 577h, 41 mg, yield 24.39%) as a yellow liquid. LCMS [M-100] + =466.2.

[0299] Step 5: Compound 577

[0300] N-(1-(5-chloro-2-(1-(4-piperidinyl)-1H-pyrazol-4-amino)pyrimidin-4-yl)-1H-pyrazol-4-yl)propane-1-sulfonamide

[0301] Trifluoroacetic acid (0.3 mL) was added to a solution of compound 577h (31.9 mg, 0.06 mmol) in dichloromethane (3 mL) at 0°C. The reaction mixture was warmed to room temperature and stirred for one hour. LCMS [M+1] + =466.1, the reaction was detected to be complete. Post-treatment: The reaction solution was concentrated to dryness to obtain a crude product, which was then purified by preparative purification to obtain the target compound as a yellow solid (Compound 577, 19.8 mg, yield 70.82%), LCMS [M+1] + =466.1.

[0302] 1 H NMR (400MHz, DMSO) δ9.99(s,1H),8.59(s,1H),8.29(d,J=50.1Hz,2H),7.93(s,1H),7.77(s,1H),7.58(s,1H),4.39(s,1 H),3.29(s,2H),3.13-3.07(m,2H),2.93(t,J=11.1Hz,2H),2.17-1.95(m,5H),1.78-1.65(m,2H),0.97(t,J=7.4Hz,3H).

[0303] Example 14: General method for the synthesis of compound 579 (TDM-181179)

[0304]

[0305] Step 1: Compound 579c

[0306] Benzyl 4-(5-((5-chloro-4-(4-(propylsulfonamido)-1H-pyrazol-1-yl)pyrimidin-2-yl)amino)thiazol-2-yl)piperidine-1-carboxylate

[0307] To a mixture of compound 579a (39 mg, 0.117 mmol), compound 579b (37 mg, 0.115 mmol), palladium acetate (5 mg, 0.023 mmol), xantphos (27 mg, 0.047 mmol) and cesium carbonate (76 mg, 0.233 mmol) was added dioxane (5 mL). The mixture was degassed under vacuum, replaced with argon several times, 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=0-25 / 75) to give a brown solid product (compound 579c, 40 mg, 55.4% yield) LCMS [M+1] + =617.

[0308] Step 2: Compound 579

[0309] N-(1-(5-chloro-2-((2-(piperidin-4-yl)thiazol-5-yl)amino)pyrimidin-4-yl)-1H-pyrazol-4-yl)propane-1-sulfonamide

[0310] Compound 579c (40 mg, 0.065 mmol) was dissolved in HBr-AcOH (2 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 yellow solid product (compound 579, TDM-181179, 7 mg, 22.3% yield). LCMS [M+1] + =483.

[0311] 1H NMR (400MHz, DMSO) δ8.68(s,1H),8.48(s,1H),8.37(s,1H),7.79(s,1H),7.41(s,1H),3.20(d,J=12.2Hz,2H),3.16(s,1H),3.12-3.07 (m,2H),2.85(t,J=11.4Hz,2H),2.52(s,1H),2.09(d,J=12.2Hz,2H),1.84(s,2H),1.71(dq,J=14.9,7.5Hz,2H),0.96(t,J=7.4Hz,3H).

[0312] Test example: Enzyme activity inhibition assay of NUAK1 / NUAK2 kinase inhibitors

[0313] 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:

[0314] 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.

[0315] Test steps:

[0316] 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).

[0317] 2. Add kinase and mix gently (the final concentration of NUAK1 is 10 nM; the final concentration of NUAK2 is 50 nM);

[0318] 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;

[0319] 4. Join 33 P-labeled ATP (unlabeled ATP / labeled ATP ratio of 25:1 or 2.5:1);

[0320] 5. Incubate at room temperature for 2 hours;

[0321] 6. Detect kinase activity using P81 filter paper combined with hotspot technology.

[0322] The IC of the test compound against NUAK1 / NUAK2 was calculated according to the above assay. 50 , see Table 1 for specific results.

[0323] IC 50 Calculations were made using the formula derived from a sigmoidal dose-response curve (variable slope):

[0324] 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.

[0325] Table 1 IC values ​​of the test compounds of this application for NUAK1 / NUAK2 50 (nM)

[0326] Test compound NUAK1 NUAK2 TDM-181134 122 622 TDM-181135 34 265 TDM-181139 474 991 TDM-181140 305 210 TDM-181146 260 602 TDM-181147 19 77 TDM-181152 977 Not tested TDM-181153 84 107 TDM-181154 58 239 TDM-181155 43 69 TDM-181156 607 Not tested TDM-181157 194 487 TDM-181158 394 Not tested TDM-181159 337 419 TDM-181169 209 495 TDM-181170 14 78 TDM-181171 9 50 TDM-181177 9 72 TDM-181179 6 42

[0327] From the results in Table 1, it can be seen that the pyrimidine amine compound of the present application has excellent inhibitory activity against NUAK1 / NUAK2 and is a dual-target small molecule kinase inhibitor. The IC of the compound against NUAK1 / NUAK2 is 50 It can reach several nM / 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.

[0328] 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.

[0329] 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.

[0330] 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 pyrimidineamine compound, characterized in that The pyrimidineamine compound is a compound represented by Formula I shown below, or a stereoisomer, tautomer, isotopic derivative, or pharmaceutically acceptable salt thereof; wherein A is phenyl or phenyl substituted by one C1-8 alkyloxy group, or a 5-membered heteroaryl group which may be substituted by p R4 groups; B is an optionally substituted 5-6 membered cycloalkyl group or an optionally substituted 5-6 membered heterocycloalkyl group; The substituents on 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; D is X is N or CH. When X is CH, H may be replaced by R5 or R6; E is -C(O)-NHR7R8, -C(O)-R7R8, -NR7-C(O)R8, -NR7-S(O)2-R8, -optionally substituted C3-8heterocycloalkyl-S(O)2-R8, -optionally substituted C3-8cycloalkyl-S(O)2-R8, R8 is optionally substituted C1-8 alkyl, optionally substituted C3-8cycloalkyl, optionally substituted C3-8cycloalkylC1-8alkyl; the substituents on the cycloalkyl or heterocycloalkyl are 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; R1 and R7 are each independently H, optionally substituted C1-8 alkyl; R2R4, R5, and R6 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; The substituents on the C1-8 alkyl group are selected from one or more of halogen, amino, hydroxy, nitro, cyano, and mercapto; n is 0, 1 or 2, and p, q, and r are 0, 1, 2 or 3 respectively.

2. The compound according to claim 1, characterized in that E is one of the following formulas:

3. The compound according to claim 1, characterized in that B is optionally substituted piperidinyl, optionally substituted piperazinyl, or optionally substituted pyrrolidinyl.

4. The compound according to claim 3, characterized in that B is one of the following formulas: R9 is 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; R 10 is H or optionally substituted C1-8 alkyl; The substituents on the C1-8 alkyl group are selected from one or more of halogen, amino, hydroxy, nitro, cyano, and mercapto; s and t are 0, 1, 2, 3 or 4 respectively, and u is 0, 1, 2 or 3.

5. The compound according to claim 4, characterized in that B is one of the following formulas:

6. The compound according to claim 1, characterized in that A is or substituted by 1 C1-8 alkyloxy group or A is one of the following groups which are optionally substituted, 7. The compound according to claim 6, characterized in that A is methoxy substituted The methoxy group is substituted at the ortho position to the amino group.

8. The compound according to claim 1, characterized in that R2 is chlorine.

9. The compound according to claim 8, characterized in that R2 is substituted at the para position of the amino group.

10. The compound according to claim 1, characterized in that The compound represented by formula I is one of the following compounds: The compound according to claim 1 , wherein the pharmaceutically acceptable salt is formate. The compound according to claim 1 , wherein the isotopic derivative is a deuterated derivative.

13. A method for preparing the compound according to any one of claims 1 to 12, comprising the following steps: Prepare a compound of formula I from a compound of formula II and a compound of formula III; or When D is When the compound of formula V is prepared from the compound of formula IV and formula II, the compound of formula I is then prepared from the compound of formula V; Optionally, the preparation process includes necessary protection and deprotection steps; in, Y is a leaving group, and the definitions of the other groups are the same as those in claims 1-12.

14. The method for preparing the compound according to claim 13, wherein Y is halogen.

15. The method for preparing the compound according to claim 14, characterized in that: Y is Cl.

16. A pharmaceutical composition, characterized in that The active ingredient is the compound according to any one of claims 1 to 12.

17. The pharmaceutical composition according to claim 16, characterized in that Contains a pharmaceutically acceptable carrier or excipient.

18. Use of the compound according to any one of claims 1 to 12 for preparing a NUAK1 or NUAK2 inhibitor.

19. Use of the compound according to any one of claims 1 to 12 for preparing a medicament, 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.

20. The use according to claim 19, 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 scars after trauma and surgery.

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