A pyrimidineamine NUAK inhibitor, its preparation method and uses

Abstract

The present application provides a pyrimidineamine compound, which is characterized in that the pyrimidineamine compound is a compound represented by Formula I, or its stereoisomer, tautomer, isotope derivative, hydrate, solvate, prodrug, and pharmaceutically acceptable salt. The pyrimidineamine compound of the present application has excellent NUAK1 / NUAK2 kinase inhibitory activity and can be used for preventing or treating the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases.

Description

Technical Field

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

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

[0003] Dysfunction of AMPK can lead to various diseases such as obesity, diabetes, inflammatory diseases and tumors. Therefore, the regulation of AMPK function by the body is very crucial. First, the activation of AMPK requires the phosphorylation of threonine (T172) at position 172 of its α subunit by upstream kinases. Currently, there are three known upstream kinases of AMPK: liver kinase B1 (LKB1), calcium / calmodulin-dependent protein kinase 2 (Ca 2+ / calmodulin-dependent PK kinase 2, CaMKK2), and transforming growth factor-β-activated kinase 1 (TAK1). Corresponding to the activation of AMPK by upstream kinases through phosphorylation of the T172 site, there are three known protein phosphatases that can inhibit the activity of AMPK by removing the phosphate group on T172, including protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), and magnesium / manganese-dependent protein phosphatase 1E (Mg 2+ - / Mn 2+ -dependent protein phosphatase 1E, PPM1E). When the cell is in a low-energy state (high AMP / ATP or ADP / ATP ratio), the KD of the α subunit and the γ subunit of AMPK are tightly cross-linked, and at the same time, the β subunit is myristoylated, ensuring that the protein phosphatase cannot access the T172 site and remains in an active state. When the cell is in a high-energy state, the KD and γ subunit loosen, and T172 is exposed to the phosphatase, resulting in the inactivation of AMPK (Steinberg GR, Nature Reviews Drug Discovery. 2019, 18:527).

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

[0005] ARKs are further divided into several ARK subfamilies according to different protein structure and function characteristics. Among them, the NUAK (Nu(novel) and AMPK-related kinase) ARK subfamily contains two members, NUAK1 (originally named ARK5) and NUAK2 (originally named sucrose nonfermenting-like / AMPK related kinase, SNARK). The amino acid sequences of the two are approximately 55% homologous. According to the amino acid sequences, the predicted molecular weights of NUAK1 and NUAK2 are 76 and 69 kDa respectively; their protein structures are very similar. The amino terminus is the kinase domain, and the carboxyl terminus is the ubiquitin-associated domain. It is not yet clear whether NUAK is also an heterotrimeric structure like AMPKs. NUAK1 and NUAK2 are expressed in most tissues. Among them, the expression of NUAK1 in organs and tissues such as the brain, skin, muscle, upper digestive tract, and endocrine is significantly higher than other parts. The organs and tissues with the highest expression of NUAK2 are the digestive tract, female reproductive system, skin, bone, brain, and endocrine, etc. Among them, the expression of NUAK2 shows more tissue specificity. It is worth mentioning that NUAK1 and other ARKs and AMPK proteins are mainly distributed in the cytoplasm, while NUAK2 is mainly distributed in the nucleus, and there are indications that NUAK2 participates in the gene expression related to metabolic stress as a transcriptional regulator (Sun X, J of Molecular Endocrinology. 2013, 51:R15).

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

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

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

[0009] Activation of NUAK1 by Akt and other protein kinases can promote the survival of tumor cells in an energy-deficient environment and protect tumor cells from apoptosis. NUAK2 also inhibits TNFα- and CD95-induced apoptosis through a similar mechanism. In addition, after activation, NUAK1 promotes the invasion and metastasis of tumor cells by upregulating matrix metalloproteinases (MMP), while NUAK2 participates in the occurrence of tumor metastasis by enhancing the motility of tumor cells (Hou X, Oncogen. 201, 30:2933; Chen Y, Cell Death and Disease. 2020, 11:712; Molina E, Cells. 10:2760; Humbert N, The EMBO Journal. 2010, 29:376). A lot of evidence shows that NUAK1 and NUAK2 play a role in tumor formation and metastasis. Compared with NUAK1, NUAK2 has a stronger effect on promoting tumor formation and metastasis. NUAK2 inhibitors should be more likely to become new targeted anti-tumor drugs than NUAK1 inhibitors.

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

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

[0012] The purpose of the present invention is to obtain effective NUAK1 / NUAK2 inhibitors, which can be used to prepare drugs for preventing or treating the following diseases: neuropsychiatric diseases (such as Parkinson's disease, Alzheimer's disease), metabolic diseases (such as diabetes, hyperlipidemia, obesity), tumors (such as liver cancer, leukemia, lymphoma, tumor metastasis), visceral fibrosis diseases (such as liver cirrhosis, renal fibrosis, pulmonary fibrosis, and sequelae of myocarditis), and skin fibrosis diseases (such as scleroderma, keloid, hypertrophic scar, or simply used to reduce scars after trauma and surgery).

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

[0014]

[0015] Wherein, A is

[0016] X is N or CH, Y is O or NR 9 ;

[0017] R 1 、R 2 、R 8 、R 9 are each independently H, optionally substituted C1-8 alkyl;

[0018] R 3 、R 4 、R7 , R 10 , R 11 Each independently is halogen, amino, hydroxyl, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino;

[0019] R 6 is optionally substituted 3-10 membered cycloalkyl, optionally substituted 3-10 membered heterocycloalkyl, wherein the substituents on the cycloalkyl or heterocycloalkyl are selected from one or more of halogen, amino, hydroxyl, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino;

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

[0021] n and r are each 0, 1 or 2, and m, p and q are each 0, 1, 2, 3 or 4.

[0022] In one group of embodiments, Y is O or NH.

[0023] In one group of embodiments, R 6 is optionally substituted 3-10 membered cycloalkyl, preferably optionally substituted 3-6 membered cycloalkyl, preferably 3-6 membered cycloalkyl substituted by halogen, more preferably R 6 is 3-6 membered cycloalkyl substituted by fluorine.

[0024] In one group of embodiments, R 6 is In one group of embodiments, R 5 is Preferably, R 5 is

[0025]

[0026] In one group of embodiments, R 5 is substituted at the ortho or meta position to Y.

[0027] In one group of embodiments, the compound represented by Formula I is Formula I-1,

[0028]

[0029] In one group of embodiments, X is N, and R 8 is H or methyl.

[0030] In one group of embodiments, the compound represented by Formula I is Formula I-2 or Formula I-3,

[0031]

[0032] In one group of embodiments, the compound represented by Formula I is one of the following compounds:

[0033]

[0034] In one group of embodiments, the pharmaceutically acceptable salt is formate.

[0035] In one group of embodiments, the isotope derivative is a deuterated compound.

[0036] On the other hand, the present application also provides a method for preparing the pyrimidineamine compound, which comprises the following steps: preparing the compound of Formula I from the compound of Formula II and the compound of Formula III;

[0037] Or

[0038] preparing the compound of Formula V from the compound of Formula IV and the compound of Formula II, and then preparing the compound of Formula I from the compound of Formula V;

[0039]

[0040] Wherein, Z is a leaving group, preferably Z is a halogen, more preferably Z is Cl, and the definitions of the other groups are as described above.

[0041] In one group of embodiments, the amino group or imino group on the six-membered nitrogen-containing heterocycle in Formula II is first protected, and after coupling with the compound of Formula III, the protecting group is removed to prepare the compound of Formula I.

[0042] In one group of embodiments, the amino group or imino group connected to R in the compound of Formula IV is first protected, optionally the amino group or imino group on the six-membered nitrogen-containing heterocycle in Formula II is protected, and then the protected compound of Formula IV is coupled with the optionally protected compound of Formula II, and the protecting group on the amino group or imino group connected to R is removed to prepare the compound of Formula V, and the compound of Formula V reacts with R 2 -COOH, and optionally the protecting group on the B ring is removed to prepare the compound of Formula I. 2 6 6 -COOH reaction, and optionally removing the protecting group on the B ring to prepare the compound of Formula I.

[0043] The present application provides a pharmaceutical composition using the pyrimidineamine compound described above in the present application as an active ingredient; the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0044] The present application also provides the use of the pyrimidineamine compound for preparing an inhibitor of NUAK1 or NUAK2.

[0045] The present application also provides the use of the pyrimidineamine compounds for the preparation of a medicament, characterized in that the compounds are used for preventing or treating the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases.

[0046] In a group of embodiments, the diseases are Parkinson's disease, Alzheimer's disease, diabetes, hyperlipidemia, obesity, liver cancer, leukemia, lymphoma, tumor metastasis, liver cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or used alone to reduce trauma and postoperative scars.

[0047] The beneficial effects of the present invention are as follows:

[0048] The present invention provides a class of pyrimidineamine compounds. In vitro kinase activity inhibition tests show that the compounds of the present invention have excellent inhibitory activity against NUAK1 or NUAK2 kinases. They can be used for preventing or treating the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases. Detailed Embodiments

[0049] The following provides a detailed description of the specific embodiments of the present invention. It should be understood that the specific embodiments described herein are only for the purpose of illustrating and explaining the present invention, and are not intended to limit the present invention.

[0050] In the ranges disclosed herein, the endpoints and any values are not limited to the exact ranges or values. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values, they can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0051] Before describing the present invention in detail, it should be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of the present invention. The scope of the present invention is only defined by the appended claims. To more fully understand the present invention described herein, the following terms are used, and their definitions are as follows. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as understood by those of ordinary skill in the art to which the present invention belongs.

[0052] Definitions

[0053] Unless otherwise specified, the following terms referred to in the present invention have the following definitions.

[0054] Features described or depicted as part of one or a group of embodiments may be used in another embodiment or another group of embodiments to yield further embodiments.

[0055] In the present invention, on the ring group It is indicated that there may be n Rs 3 , m Rs 4 , p Rs 7 , q Rs 10 , r Rs 11 connected at any possible position on the said ring group.

[0056] In the present invention, at the position of in some substituents indicates the connection site. For the A group, it represents being connected to an amino group above and a nitrogen-containing six-membered heterocycle below.

[0057] Unless otherwise specified, the term "optionally substituted" means that the hydrogen on the group to be substituted is unsubstituted or one or more substitutable sites on the substituting group are independently substituted by substituents, and the substituents are independently selected from one or more of deuterium, halogen, amino, hydroxy, nitro, cyano, mercapto, oxo, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino; the substituents on the C1-8 alkyl are selected from one or more of deuterium, halogen, amino, hydroxy, nitro, cyano, mercapto; when the substituent is selected from "oxo", it means that two hydrogen atoms at the same substitution position are replaced by an oxygen atom.

[0058] The term "cycloalkyl" denotes a saturated monocyclic, bicyclic or tricyclic system containing 3 to 10 carbon atoms, wherein the monocyclic, bicyclic or tricyclic system does not contain an aromatic ring. The bicyclic group includes bridged ring groups, spiro ring groups, fused ring groups, etc. Preferably, it contains 3 to 10 carbon atoms (C3-10 cycloalkyl), more preferably 3 to 8 carbon atoms (C3-8 cycloalkyl), 3 to 6 carbon atoms (C3-6 cycloalkyl), 4 to 6 carbon atoms (C4-6 cycloalkyl), 5 to 6 carbon atoms (C5-6 cycloalkyl). Examples include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, etc.

[0059] The term "heterocycloalkyl" refers to a saturated monocyclic, bicyclic or polycyclic cyclic hydrocarbon group, preferably containing 3-10 ring atoms, wherein 1, 2, 3 or more ring atoms are selected from N, O or S, and the remaining ring atoms are C, including bridged ring groups, spiro ring groups, fused ring groups, etc. Preferably, it contains 3-8 ring atoms (3-8 membered heterocycloalkyl), or 3-6 ring atoms (3-6 membered heterocycloalkyl), or 4-6 ring atoms (4-6 membered heterocycloalkyl), or 5-6 ring atoms (5-6 membered heterocycloalkyl). The number of heteroatoms is preferably 1-4, more preferably 1-3 (i.e., 1, 2 or 3). The heterocycloalkyl can be a 5-6 membered monocyclic heterocycloalkyl containing 1-2 N atoms. Examples of heterocycloalkyl include pyrrolidinyl, imidazolidinyl, tetrahydrofuryl, piperidinyl, piperazinyl, pyranyl, aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, oxane, morpholinyl, thiomorpholinyl, dioxolanyl, dithiane, oxazolidinyl, thiazolidinyl, pyrazolidinyl, imidazolidine, etc.

[0060] The term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, preferably a straight-chain or branched-chain group containing 1-8 carbon atoms (C1-8 alkyl) (the number of carbon atoms is between 1 and 8, specifically 1, 2, 3, 4, 5, 6, 7 or 8), more preferably containing 1-6 carbon atoms (i.e., C1-6 alkyl, the number of carbon atoms is between 1 and 6, specifically 1, 2, 3, 4, 5 or 6). Examples include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, etc.

[0061] The terms "alkyloxy", "alkylthio", "alkylamino" refer to -O-alkyl, -S-alkyl, -NH-alkyl or dialkylamino respectively, and the alkyl is as defined above. Representative examples include but are not limited to methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, tert-butoxy, etc.; methylthio, ethylthio, propylthio, isopropylthio, butylthio, 1-methylpropylthio, 2-methylpropylthio, tert-butylthio, etc.; methylamino, ethylamino, propylamino, dimethylamino, diethylamino, dipropylamino, methylethylamino, etc.

[0062] The term "halogen" refers to F, Cl, Br, I.

[0063] The active compounds described in the present invention are construed to include the compounds and their stereoisomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts thereof. The stereoisomers, tautomers, isotopic derivatives, hydrates, solvates, prodrugs, isotopic derivatives, or pharmaceutically acceptable salts of the compounds are obtained by conventional techniques in the art and exhibit the same or similar effects in vivo and in vitro through substantially the same mechanism of action as the compounds.

[0064] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including configurational isomers and conformational isomers. Among them, configurational isomers include geometric isomers (or cis-trans isomers) and optical isomers (including enantiomers and diastereomers). Geometric isomers may exist in the present compounds. Optical isomers refer to substances with identical molecular structures, similar physicochemical properties, but different optical activities. The compounds of the present invention may contain asymmetrically substituted carbon atoms in the R or S configuration, where the terms "R" and "S" are defined as in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem. (1976) 45, 13 - 10. Compounds with asymmetrically substituted carbon atoms (with an equal number of R and S configurations) are racemic at those carbon atoms. Atoms with an excess of one configuration (relative to the other) result in a higher quantity of that configuration, preferably in an excess of approximately 85% - 90%, more preferably in an excess of approximately 9% - 99%, and even more preferably in an excess greater than approximately 99%. Accordingly, the present invention includes racemic mixtures, relative and absolute optical isomers, and mixtures of relative and absolute optical isomers.

[0065] The term "tautomer" refers to structural isomers with different energies that can be interconverted through a low energy barrier. If tautomerism is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also known as proton transfer tautomers) include interconversions through proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions through the reorganization of some bonding electrons.

[0066] The term "isotopic derivative" means that the compounds of the present invention may exist in isotopically labeled or enriched forms, containing one or more atoms whose atomic weights or mass numbers are different from those of the atoms found in the greatest abundance in nature. Isotopes can be radioactive or non-radioactive isotopes. Isotopes of atoms such as hydrogen, carbon, phosphorus, sulfur, fluorine, chlorine, and iodine include, but are not limited to: 2 H, 3H, 13 C, 14 C, 15 N, 18 O, 32 P, 35 S, 18 F, 36 Cl and 125 I. Compounds containing these and / or other isotopes of these atoms are within the scope of the present invention. The isotopically labeled compounds of the present invention can be prepared by general methods well known to those of ordinary skill in the art.

[0067] The term "hydrate" refers to an association formed by one or more water molecules with the compounds of the present invention.

[0068] The term "solvate" refers to an association formed by one or more solvent molecules with the compounds of the present invention.

[0069] The term "prodrug" refers to a derivative of an active drug that is designed to improve certain defined, undesirable physical or biological properties. Physical properties are generally related to solubility (excessive or insufficient lipid or water solubility) or stability, while problematic biological characteristics include too rapid metabolism or poor bioavailability, which may in turn be related to physicochemical properties.

[0070] The term "pharmaceutically acceptable salt" refers to salts that are suitable, within the scope of sound medical judgment, for contact with the tissues of mammals, particularly humans, without undue toxicity, irritation, allergic response, etc., and that are commensurate with a reasonable benefit / risk ratio. If the compound is basic, pharmaceutically acceptable salts include salts prepared from inorganic acids and also include salts prepared from organic acids. If the compound is acidic, pharmaceutically acceptable salts include salts prepared from inorganic bases and / or organic bases.

[0071] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent, or emulsifying agent, etc., that has been approved by the relevant government regulatory authorities for use in humans or livestock.

[0072] As used herein, the term "treatment" refers to any administration of a therapeutic agent according to a therapeutic regimen that achieves a desired effect, i.e., partially or completely alleviating, ameliorating, relieving, inhibiting, delaying the onset, reducing the severity, and / or reducing the incidence of one or more symptoms or characteristics of a particular disease, disorder, and / or condition; in some embodiments, the administration of a therapeutic agent according to a therapeutic regimen is associated with the achievement of the desired effect. Such treatment can be directed to a subject who does not exhibit the relevant disease, disorder, and / or condition and / or to a subject who exhibits only early signs of the disease, disorder, and / or condition. Alternatively or additionally, such treatment can be directed to a subject who exhibits one or more of the identified signs of the relevant disease, disorder, and / or condition. In some embodiments, the treatment can be directed to a subject who has been diagnosed with the relevant disease, disorder, and / or condition. In some embodiments, the treatment can be directed to a subject known to have one or more risk factors that are statistically associated with an increased risk of developing the relevant disease, disorder, and / or condition.

[0073] According to the present invention, the drug prepared in the pharmaceutical use described in the present application may contain, in addition to the pyrimidineamine compound of the present invention as an active ingredient, other agents that can be used to prevent or treat related diseases as another active ingredient. When the drug contains multiple active ingredients, the active ingredients can be administered simultaneously, sequentially, or separately according to the judgment of a physician.

[0074] Hereinafter, the effects of specific compounds of the present invention will be described in detail by way of examples.

[0075] Example

[0076] General method for synthesizing compound 518 (TDM-181118) in Example 1

[0077]

[0078] Step 1: Compound 518c

[0079] tert-Butyl (S)-4-(4-((4-(4-(2,2-difluorocyclopropane-1-carboxamido)phenyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0080] To a mixture of compound 518a (20 mg, 0.065 mmol), compound 518b (26 mg, 0.097 mmol), palladium acetate, xantphos (37 mg, 0.065 mmol) and cesium carbonate (42 mg, 0.129 mmol) was added dioxane (2 mL). The mixture was degassed under vacuum, purged with argon several times, heated to 100 °C and stirred for 2 h. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 27 / 73 with 10% methanol) to give the yellow solid product (compound 518c, 17.3 mg, yield 49.3%). LCMS [M+1] + = 540.

[0081] Step 2: Compound 518

[0082] (S)-2,2-difluoro-N-(4-(2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)cyclopropane-1-carboxamide

[0083] To a solution of compound 518c (17.3 mg, 0.028 mmol) in methanol (0.5 mL) and dichloromethane (2 mL) was added 4 M hydrochloric acid dioxane solution (0.07 mL). The mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC (formic acid) to give the white solid product (compound 518, TDM-181118, 13.5 mg, yield 33%). LCMS [M+1] + = 440.

[0084] 1 H NMR (400 MHz, DMSO) δ 10.76 (s, 1H), 9.47 (s, 1H), 8.45 (d, J = 5.1 Hz, 1H), 8.35 (s, 1H), 8.13 (d, J = 8.7 Hz, 2H), 7.98 (s, 1H), 7.77 (d, J = 8.5 Hz, 2H), 7.62 (s, 1H), 7.24 (d, J = 5.2 Hz, 1H), 4.33 (s, 1H), 3.21 (d, J = 12.5 Hz, 2H), 2.93 - 2.75 (m, 3H), 2.13 - 1.87 (m, 6H).

[0085] N-(3-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)oxy)phenyl)-2,2-difluorocyclopropane-1-carboxamide (TDM-181120), a white solid (19.5 mg, yield 13.7%), was synthesized in a similar manner to Example 1.

[0086]

[0087]

[0088] Example 2 General method for synthesizing compound 527 (TDM-181127)

[0089]

[0090] Step 1: Compound 527c 2,5-dichloro-4-(3-nitrophenoxy)pyrimidine

[0091] Potassium carbonate (904 mg, 6.542 mmol) was added to a solution of compound 527a (1000 mg, 5.452 mmol) and compound 527b (758 mg, 5.452 mmol) in N,N-dimethylformamide (40 mL). The mixture was stirred at room temperature for 2 hours. Then the mixture was added to water (250 mL) and extracted with ethyl acetate (80 mL * 3). The combined organic layers were washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was triturated with dichloromethane and methanol and filtered to obtain a white solid product (compound 527c, 1.34 g, 85.9% yield). LCMS [M+1] + = 286.

[0092] Step 2: Compound 527d

[0093] 3-((2,5-Dichloropyrimidin-4-yl)oxy)aniline

[0094] Ammonium chloride (145 mg, 2.622 mmol) and iron powder (148 mg, 2.622 mmol) were added to a mixed solution of compound 527c (150 mg, 0.524 mmol) in tetrahydrofuran (10 mL) and water (10 mL). The mixture was heated to 65 °C and stirred overnight. Then the mixture was filtered and the filtrate was concentrated under reduced pressure to remove tetrahydrofuran. The residue was neutralized with aqueous sodium bicarbonate, extracted with ethyl acetate (30 mL * 3), the combined organic layers were washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 1 / 99 with 10% methanol) to obtain a white solid product (compound 527d, 80 mg, 59.6% yield). LCMS [M+1] + = 256.

[0095] Step 3: Compound 527f

[0096] N-(3-((2,5-Dichloropyrimidin-4-yl)oxy)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0097] To a solution of compound 527d (80 mg, 0.312 mmol), compound 527e (57 mg, 0.469 mmol) and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (178 mg, 0.469 mmol) in N,N-dimethylformamide (8 mL) was added N,N-diisopropylethylamine (89 mg, 0.469 mmol). The mixture was heated to 50 °C and stirred for 2 h. The mixture was added with water (80 mL), then extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 1 / 99 with 10% methanol) to give a yellow solid product (compound 527f, 70 mg, 62.3% yield). LCMS [M+1] + = 360.

[0098] Step 4: Compound 527h

[0099] tert-Butyl 4-(4-((5-chloro-4-(3-(2,2-difluorocyclopropane-1-carboxamido)phenoxy)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0100] To a mixture of compound 527f (70 mg, 0.194 mmol), compound 527g (62 mg, 0.233 mmol), palladium acetate (8.8 mg, 0.039 mmol), xantphos (45 mg, 0.078 mmol) and cesium carbonate (126 mg, 0.388 mmol) was added dioxane (7 mL). The mixture was degassed under vacuum, purged with argon several times, heated to 100 °C and stirred for 2 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (dichloromethane / methanol = 0 - 30 / 70 with 10% methanol) to give a yellow solid product (compound 527h, 55 mg, 48% yield). LCMS [M+1] + = 590.

[0101] Step 5: Compound 527

[0102] N-(3-((5-chloro-2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)oxy)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0103] To a solution of compound 527h (55 mg, 0.093 mmol) in methanol (1 mL) and dichloromethane (5 mL) was added dioxane hydrochloride solution (0.35 mL). The mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to give the white solid product (compound 527, TDM-181127, 25 mg, 54.9% yield). LCMS [M+1] + = 490.

[0104] 1 H NMR (400 MHz, MeOD) δ 8.55 (s, 1H), 8.26 (s, 1H), 7.99 (s, 1H), 7.53 (s, 1H), 7.23 (d, J = 30.0 Hz, 2H), 7.10–6.81 (m, 2H), 4.01 (s, 1H), 3.52 - 3.40 (m, 2H), 3.15 (dd, J = 20.8, 7.9 Hz, 2H), 2.71 (ddd, J = 13.2, 10.8, 7.8 Hz, 1H), 2.20 - 1.81 (m, 6H).

[0105] General procedure for the synthesis of compound 530 (TDM-181130) in Example 3

[0106]

[0107] Step 1: Compound 530c

[0108] 2,5-Dichloro-N-methyl-N-(3-nitrophenyl)pyrimidin-4-amine

[0109] To a solution of compound 530b (730 mg, 4.8 mmol) in N,N-dimethylformamide (40 mL) at 0 °C was added sodium hydride (384 mg, 9.6 mmol). The mixture was allowed to warm to room temperature and stirred for one hour, then compound 530a (1232 mg, 6.72 mmol) was added, and the mixture was stirred at room temperature for two hours. The reaction was monitored to completion. Workup: The reaction mixture was poured into water, and the aqueous phase was extracted three times with ethyl acetate (3 * 150 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The crude product obtained was purified by column chromatography [eluent: EA / PE = 0 - 10%] to give the yellow solid target compound (compound 530c, 708.6 mg, 49.35% yield), LCMS [M+1] + = 299, 301.

[0110] Step 2: Compound 530d

[0111] N1-(2,5-Dichloropyrimidin-4-yl)-N1-methylbenzene-1,3-diamine

[0112] To a solution of compound 530c (605.5 mg, 2.02 mmol) in tetrahydrofuran (60 mL) and water (60 mL) were added iron powder (565.3 mg, 10.12 mmol) and ammonium chloride (541.3 mg, 10.12 mmol). The reaction mixture was heated to 65 °C and stirred for 8 hours. The reaction was monitored to completion. Work-up: The reaction mixture was cooled to room temperature and filtered. The filter cake was washed with ethyl acetate. The filtrate was extracted with ethyl acetate three times. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography [eluent: (methylene chloride / methanol = 10:1) / DCM = 0 - 40%] to give the yellow solid target compound (compound 530d, 603 mg, yield 95%), LCMS [M+1] + = 269, 271.

[0113] Step 3: Compound 530f

[0114] (S)-N-(3-((2,5-Dichloropyrimidin-4-yl)(methyl)amino)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0115] To a solution of compound 530d (537.4 mg, 2 mmol) in pyridine (20 mL) was added compound 530e (244 mg, 2 mmol). After cooling the reaction mixture to 0 °C, phosphorus oxychloride (0.28 mL, 3 mmol) was added. The reaction mixture was stirred at this temperature for one hour. The reaction was monitored to completion. Work-up: The reaction mixture was poured into ice water (30 mL). The aqueous phase was extracted with ethyl acetate (2 * 50 mL) twice. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography, eluent: EA / PE = 0 - 40%, to give the yellow solid target compound (compound 530f, 273.5 mg, yield 36.6%), LCMS [M+1] + = 373, 375.

[0116] Step 4: Compound 530

[0117] (S)-N-(3-(5-Chloro-2-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)(methyl)amino)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0118] To a solution of compound 530f (52 mg, 0.14 mmol) in n-butanol (5 mL) was added compound 530g (31 mg, 0.14 mmol) and p-toluenesulfonic acid (53.3 mg, 0.28 mmol). The reaction mixture was heated to 110 °C and stirred for 4 h. The reaction was monitored and found to be complete. Work-up: The reaction mixture was poured into water, and the aqueous phase was extracted twice with ethyl acetate (2 × 150 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified by preparative TLC to give the yellow solid target compound (compound 530, 15.4 mg, yield 19.7%), LCMS [M+1] + = 558.

[0119] 1 H NMR (400 MHz, DMSO) δ 10.46 (s, 1H), 7.98 (s, 1H), 7.87 (s, 1H), 7.73 (d, J = 8.7 Hz, 1H), 7.40 (dd, J = 8.0, 5.0 Hz, 2H), 7.31 (t, J = 8.0 Hz, 1H), 6.91 (d, J = 9.1 Hz, 1H), 6.63 (d, J = 2.5 Hz, 1H), 6.47 (dd, J = 8.8, 2.5 Hz, 1H), 3.83 (s, 3H), 3.36 (s, 3H), 3.15 - 3.07 (m, 4H), 2.77 (ddd, J = 13.6, 10.9, 8.1 Hz, 1H), 2.47 - 2.43 (m, 4H), 2.22 (s, 3H), 2.06 - 1.90 (m, 2H).

[0120] (S)-N-(3-(5-chloro-2-(4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)(methyl)amino)phenyl)-2,2-difluorocyclopropane-1-carboxamide (compound 531, TDM-181131) was synthesized in a similar manner to Example 3 as an off-white solid (9.4 mg, yield 10%).

[0121]

[0122] General procedure for the synthesis of compound 532 in Example 4 (TDM-181132)

[0123]

[0124] Step 1: Compound 532c

[0125] 2,5-dichloro-4-(2-nitrophenoxy)pyrimidine

[0126] To a solution of compound 532b (421 mg, 3.03 mmol) in N,N-dimethylformamide (20 mL) was added compound 532a (550 mg, 3.03 mmol) and potassium carbonate (502 mg, 3.63 mmol). The reaction mixture was stirred at room temperature for two hours and the reaction was detected to be complete. Work-up: The reaction mixture was poured into water, and the aqueous phase was extracted with ethyl acetate (3 * 50 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product which was purified by column chromatography [eluent: PE / EA = 0 - 15%] to give the white solid target compound (compound 532c, 800 mg, yield 92.4%), LCMS [M+1] + = 286, 288.

[0127] Step 2: Compound 532d

[0128] 2-((2,5-Dichloropyrimidin-4-yl)oxy)aniline

[0129] To a solution of compound 532c (1.54 g, 5.4 mmol) in tetrahydrofuran (150 mL) and water (150 mL) was added iron powder (1.51 g, 27 mmol) and ammonium chloride (1.44 g, 27 mmol). The reaction mixture was heated to 65 °C and stirred for 3 hours, and the reaction was detected to be complete. Work-up: The reaction mixture was cooled to room temperature and filtered, and the filtrate was concentrated. Water and ethyl acetate (20 mL) were added and the mixture was extracted three times. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product which was purified by column chromatography [eluent: (dichloromethane / methanol = 10:1) / DCM = 0 - 40%] to give the yellow solid target compound (compound 532d, 476 mg, yield 34%), LCMS [M+1] + = 256, 258.

[0130] Step 3: Compound 532f

[0131] N-(2-((2,5-Dichloropyrimidin-4-yl)oxy)phenyl)cyclopropanecarboxamide

[0132] To a solution of compound 532d (408 mg, 1.59 mmol) in pyridine (25 mL) was added compound 532e (194.5 mg, 1.59 mmol). After cooling the reaction mixture to 0 °C, phosphorus oxychloride (0.22 mL, 2.39 mmol) was added, and the mixture was stirred at the same temperature for one hour. The reaction was monitored and found to be complete. Work-up: The reaction mixture was poured into ice-water (30 mL), and the aqueous layer was extracted twice with ethyl acetate (2 × 50 mL). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography using an eluent of EA / PE = 0 - 40%, affording the target compound as a yellow solid (compound 532f, 396.8 mg, yield 59%). LCMS [M+1] + = 325.

[0133] Step 4: Compound 532

[0134] N-(2-(5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)oxy)phenyl)cyclopropanecarboxamide

[0135] To a 100 mL three-necked flask were added compound 532f (30 mg, 0.12 mmol), compound 532g (30.5 mg, 0.14 mmol), palladium(II) 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 mixture was purged with argon several times and then stirred at 100 °C for two hours. The reaction was monitored and found to be complete. Work-up: The reaction mixture was poured into ice-water (30 mL), and the aqueous layer was extracted twice with ethyl acetate (2 × 50 mL). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by preparative TLC, affording the target compound as a yellow solid (compound 532, 14.3 mg, yield 19.7%). LCMS [M+1] + = 255, 509.

[0136] 1 H NMR (400 MHz, DMSO) δ 8.30 (s, 1H), 8.07 (s, 1H), 7.72 - 7.64 (m, 1H), 7.56 (s, 2H), 7.23 (d, J = 5.9 Hz, 3H), 6.58 (d, J = 2.2 Hz, 1H), 6.32 (d, J = 8.4 Hz, 1H), 3.77 (s, 3H), 3.08 (s, 4H), 2.45 (s, 4H), 2.22 (s, 3H), 1.76 (d, J = 5.0 Hz, 1H), 1.00 - 0.86 (m, 4H).

[0137] General method for synthesizing compound 560 (TDM - 181160) in Example 5

[0138]

[0139] Step 1: Compound 560c

[0140] 2,5 - Dichloro - 4-(2 - nitrophenoxy)pyrimidine

[0141] To a solution of compound 560b (421 mg, 3.03 mmol) in N,N - dimethylformamide (20 mL) was added compound 560a (550 mg, 3.03 mmol) and potassium carbonate (502 mg, 3.63 mmol). The reaction mixture was stirred at room temperature for 2 hours and the reaction was detected to be complete. Work - up: The reaction mixture was poured into water, and extracted three times with ethyl acetate (3 * 50 mL) for liquid - liquid separation. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered by suction, and concentrated to dryness. The obtained crude product was purified by column chromatography with the eluent: PE / EA = 0 - 15%, to obtain the white solid target compound (compound 560c, 800 mg, yield 92.4%), LCMS [M + 1] + = 286, 288.

[0142] Step 2: Compound 560d

[0143] 2 - ((2,5 - Dichloropyrimidin - 4 - yl)oxy)aniline

[0144] To a solution of compound 560c (1.54 g, 5.4 mmol) in tetrahydrofuran (150 mL) and water (150 mL) was added iron powder (1.51 g, 27 mmol) and ammonium chloride (1.44 g, 27 mmol). The reaction mixture was heated to 65 °C and stirred for 3 hours, and the reaction was detected to be complete. Work - up: The reaction mixture was cooled to room temperature, filtered by suction, the filter cake was washed with ethyl acetate, the filtrate was extracted three times with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered by suction, and concentrated to dryness. The obtained crude product was purified by column chromatography with the eluent: PE / EA = 0 - 50%, to obtain the yellow solid target compound (compound 560d, 476 mg, yield 34%), LCMS [M + 1] + = 256, 258.

[0145] Step 3: Compound 560f

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

[0147] To a solution of compound 560d (408 mg, 1.59 mmol) in pyridine (25 mL) was added compound 560e (194.5 mg, 1.59 mmol). After cooling the reaction mixture to 0 °C, phosphorus oxychloride (0.22 mL, 2.39 mmol) was added, and the mixture was kept warm for 30 minutes. The reaction was monitored to completion. Work-up: The reaction mixture was poured into ice water (30 mL). The aqueous phase was extracted twice with ethyl acetate (2 × 50 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product obtained was purified by column chromatography using an eluent of PE / EA = 0 - 20%, to give the yellow solid target compound (compound 560f, 396.8 mg, yield 59%). LCMS [M+1] + = 360, 362.

[0148] Step 4: Compound 560

[0149] (S)-N-(2-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)oxy)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0150] To a solution of compound 560f (339 mg, 0.94 mmol) in n-butanol (24 mL) were added compound 560g (208 mg, 0.94 mmol) and p-toluenesulfonic acid monohydrate (357 mg, 1.88 mmol). The reaction mixture was heated to 110 °C and stirred for 4 hours. The reaction was monitored to completion. Work-up: The reaction mixture was poured into water. The aqueous phase was extracted with ethyl acetate (2 × 150 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product obtained was purified by column chromatography using an eluent of (dichloromethane:methanol = 10:1 / DCM = 0 - 50%). The crude product was further purified by preparative purification to give the white solid target compound (compound 560, 12.7 mg, yield 17%). LCMS [M+1] + = 545.2.

[0151] 11H NMR (400 MHz, DMSO) δ 8.38 (s, 1H), 8.06 (s, 1H), 7.70 (dd, J = 7.0, 3.0 Hz, 1H), 7.55 (d, J = 12.4 Hz, 2H), 7.26 (dd, J = 6.3, 3.7 Hz, 3H), 6.58 (d, J = 2.5 Hz, 1H), 6.31 (d, J = 8.7 Hz, 1H), 3.77 (s, 3H), 3.11 - 3.07 (m, 4H), 2.99 - 2.89 (m, 1H), 2.49 - 2.44 (m, 4H), 2.25 (s, 3H), 2.13 (dt, J = 9.4, 5.8 Hz, 1H), 2.02 (td, J = 14.1, 7.9 Hz, 1H).

[0152] General method for synthesizing compound 568 in Example 6 (TDM - 181168)

[0153]

[0154] Step 1: Compound 568b

[0155] N1-(2,5 - Dichloropyrimidin - 4 - yl)-N1 - methylbenzene - 1,2 - diamine

[0156] To a solution of compound 568a (243 mg, 0.813 mmol) in tetrahydrofuran (2.5 mL) and water (2.5 mL) was added iron powder (28 mg, 0.502 mmol) and ammonium chloride (26.8 mg, 0.502 mmol). The mixture was heated to 65 °C and stirred. The mixture was filtered and the filtrate was concentrated under reduced pressure to remove some tetrahydrofuran. The residue was neutralized with aqueous sodium carbonate solution, 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 (100% dichloromethane) to give a yellow solid product (compound 568b, 112.3 mg, 45.7% yield). LCMS [M + 1] + = 269, 271.

[0157] Step 2: Compound 568d

[0158] (S)-N-(2 - ((2,5 - Dichloropyrimidin - 4 - yl)(methyl)amino)phenyl)-2,2 - difluorocyclopropane - 1 - carboxamide

[0159] Under an argon atmosphere at 0 °C, phosphorus oxychloride (96 mg, 0.626 mmol) was added to a solution of compound 568b (112.3 mg, 0.417 mmol) and compound 568c (61 mg, 0.501 mmol) in pyridine (5 mL). The mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (100% dichloromethane) to give a yellow solid product (compound 568d, 141 mg, 90.6% yield). LCMS [M+1] + = 373, 375.

[0160] Step 3: Compound 568

[0161] (S)-N-(2-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)(methyl)amino)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0162] p-Toluenesulfonic acid monohydrate (71.5 mg, 0.376 mmol) was added to a solution of compound 568d (70 mg, 0.188 mmol) and compound 568e (53.8 mg, 0.288 mmol) in n-butanol (5 mL). The mixture was heated to 110 °C and stirred for 6 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (25 mL × 3). The combined organic layers were washed with brine, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (formic acid) to give a white solid product (compound 568, TDM-181168, 73.9 mg, 58% yield) LCMS [M+1] + = 528.

[0163] 1 H NMR (400 MHz, DMSO) δ 9.86 (s, 1H), 9.23 (s, 1H), 8.16 (s, 1H), 8.05 (d, J = 8.2 Hz, 1H), 7.93 (s, 1H), 7.59 (d, J = 9.0 Hz, 2H), 7.32 - 7.23 (m, 1H), 7.09 (d, J = 4.1 Hz, 2H), 6.89 (s, 2H), 3.23 (s, 3H), 3.20–3.11 (m, 1H), 3.09 - 3.02 (m, 4H), 2.49 - 2.43 (m, 4H), 2.23 (s, 3H), 2.09 - 1.87 (m, 2H).

[0164] Test Example: Detection of Enzyme Activity Inhibition of NUAK1 / NUAK2 Kinase Inhibitor

[0165] The inhibitory activity of the pyrimidine amine small molecules involved in this application against NUAK1 and NUAK2 kinases was tested using a kinase activity assay based on the P81 filter paper binding hot spot technology (Nat Biotechnol. 2011, 29: 1039), which is briefly described as follows:

[0166] The test buffer system contains 20 mM Hepes (pH 7.5), 10 mM MgCl 2 , 1 mM EGTA, 0.01% Brij35 (L23 polyoxyethylene lauryl ether), 0.02 mg / ml BSA (bovine serum albumin), 0.1 mM Na 3 VO 4 , 2 mM DTT, 1% DMSO.

[0167] Test steps:

[0168] 1. Prepare a 20 μM substrate solution with freshly prepared test buffer. The substrate is the CHKtide polypeptide fragment (Sanchez Y. Science. 1997, 277: 1497 - 1501);

[0169] 2. Add the kinase and mix gently (the final concentration of NUAK1 is 10 nM; the final concentration of NUAK2 is 50 nM);

[0170] 3. Use acoustic liquid handling technology (Echo550; nanoliter scale) to add the test compound dissolved in 100% DMSO to the above kinase reaction mixture and incubate at room temperature for 5 minutes;

[0171] 4. Add 33 P - labeled ATP (the ratio of unlabeled ATP to labeled ATP is 25:1 or 2.5:1);

[0172] 5. Incubate at room temperature for 2 hours;

[0173] 6. Detect the kinase activity by the P81 filter paper binding hot spot technology.

[0174] Calculate the IC 50 of the test compound against NUAK1 / NUAK2. For the specific results, see Table 1.

[0175] IC 50 The calculation uses the formula obtained from a sigmoidal dose - response curve (variable slope):

[0176] Y = Bottom + (Top - Bottom) / (1 + 10^((LogEC50 - X)*slope)), where X is the Log value of the compound concentration, Y is the response (inhibition rate of kinase activity), and Y increases from bottom to top along the sigmoidal curve as the concentration increases.

[0177] Table 1 IC of the test compounds of the present application against NUA K1 / NUA K2 50 (nM)

[0178] Test compound NUAK1 NUAK2 TDM-181120 21 311 TDM-181127 161 352 TDM-181131 81 635 TDM-181132 24 37 TDM-181160 4 13

[0179] As can be seen from the results in Table 1, the pyrimidinamine compounds of the present application have excellent inhibitory activities against both NUA K1 / NUA K2, and are a dual-target small molecule kinase inhibitor. The IC of the compounds against NUA K1 / NUA K2 50 can reach several nM or dozens of nM. Therefore, it has been proven through the above experiments that the pyrimidinamine compounds of the present application can be used as NUA K1 / NUA K2 inhibitors. The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0180] In addition, it should be noted that, in the case of no conflict, the various specific technical features described in the above specific embodiments can be combined in any suitable manner. To avoid unnecessary repetition, the present invention will not separately describe various possible combination manners.

[0181] Furthermore, any combination can be made between various different embodiments of the present invention, as long as it does not violate the idea of the present invention, and it should also be regarded as the content disclosed by the present invention.

Claims

1. A pyrimidineamine compound, characterized in that, the pyrimidineamine compound is a compound represented by Formula I-1, or a stereoisomer, tautomer, isotope derivative thereof, and a pharmaceutically acceptable salt; where A is X is N or CH, and Y is O or NR 9 ; R 1 、R 2 、R 8 、R 9 Each independently is H, an optionally substituted C1-8 alkyl group; R 3 、 R 4 、 R 7 、 R 10 、 R 11 Each independently represents a halogen, amino group, hydroxyl group, nitro group, cyano group, mercapto group, optionally substituted C1-8 alkyl group, optionally substituted C1-8 alkoxy group, optionally substituted C1-8 alkylthio group, optionally substituted C1-8 alkylamino group; R 6 is an optionally substituted 3- to 10-membered cycloalkyl group, wherein the substituents on the cycloalkyl group 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; the substituent on the C1-8 alkyl is selected from one or more of halogen, amino, hydroxyl, nitro, cyano, and mercapto; n and r are each 0, 1, or 2, and m, p, and q are each 0, 1, 2, 3, or 4; The pyrimidineamine compound is not 2. The compound according to claim 1, characterized in that Y is O or NH.

3. The compound according to claim 1, characterized in that, R 6 is an optionally substituted 3- to 6-membered cycloalkyl group.

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

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

6. The compound according to claim 3, characterized in that, R 6 For 7. The compound according to claim 6, characterized in that, R 5 For 8. The compound according to claim 7, characterized in that, R 5 For 9. The compound according to any one of claims 1-8, characterized in that, R 5 is substituted at the ortho- or meta-position to Y.

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

11. The compound according to claim 10, characterized in that the compound represented by Formula I-1 is 12. The compound according to claim 1, characterized in that, the compound represented by Formula I is one of the following compounds:

13. For the compound according to claim 1 or 12, the pharmaceutically acceptable salt is formate.

14. For the compound according to claim 1 or 12, the isotope derivative is a deuterated compound.

15. A method for preparing the compound according to any one of claims 1-14, which comprises the following steps: preparing a compound of Formula I from a compound of Formula II and a compound of Formula III; or preparing a compound of Formula V from a compound of Formula IV and a compound of Formula II, and then preparing a compound of Formula I from the compound of Formula V; optionally, necessary protection and deprotection steps are included in the preparation process; wherein, Z is a leaving group, and the definitions of the other groups are the same as those defined in claims 1-14.

16. The preparation method according to claim 15, characterized in that, the Z is halogen.

17. The preparation method according to claim 16, characterized in that, the Z is Cl.

18. A pharmaceutical composition, characterized in that uses the compound according to any one of claims 1-14 as an active ingredient.

19. The pharmaceutical composition according to claim 18, characterized in that it contains a pharmaceutically acceptable carrier or excipient.

20. Use of the compound according to any one of claims 1-14 for preparing a NUAK1 or NUAK2 inhibitor.

21. Use of the compound according to any one of claims 1-14 for preparing a drug, characterized in that the compound is used for preventing or treating the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases.

22. The use according to claim 21, characterized in that, The disease is Parkinson's disease, Alzheimer's disease, diabetes, hyperlipidemia, obesity, liver cancer, leukemia, lymphoma, tumor metastasis, liver cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or is used alone to reduce scars after trauma and surgery.

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