Pyrimidinamine NUAK inhibitor as well as preparation method therefor and use thereof

Abstract

Provided in the present application are pyrimidinamine compounds, characterized in that the pyrimidinamine compounds are compounds represented by formula I, or stereoisomers, tautomers, isotope derivatives, hydrates, solvates, prodrugs and pharmaceutically acceptable salts thereof. The pyrimidinamine compounds of the present application have excellent NUAK1 / NUAK2 kinase inhibitory activity, and thus can be used for preventing or treating the following diseases by means of inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases.

Description

A pyrimidineamine NUAK inhibitor and its preparation method and use

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application No. 202310866091.6 filed on July 13, 2023, the entire contents of which are incorporated herein by reference. Technical Field

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

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

[0005] AMPK dysfunction can lead to a variety of diseases such as obesity, diabetes, inflammatory diseases and tumors, so the body's regulation of AMPK function is very critical. First, the activation of AMPK requires upstream kinases to phosphorylate the threonine (T172) at position 172 of its α subunit. Currently, there are three known upstream kinases of AMPK: liver kinase B1 (LKB1), calcium ion / calmodulin-dependent protein kinase 2 (Ca2+), and calcium ion / calmodulin-dependent protein kinase 2 (Ca3+). 2+ / calmodulin-dependent PK kinase 2, CaMKK2), and transforming growth factor-β-activated kinase 1 (TAK1). Corresponding to the activation of AMPK by upstream kinases through phosphorylation of T172 site, three protein phosphatases are known to inhibit AMPK activity by removing the phosphate group on T172, including protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), and magnesium / manganese-dependent protein phosphatase 1E (Mg / Mn-dependent protein phosphatase 1E). 2+ - / Mn 2+ -dependent protein phosphatase 1E, PPM1E). When cells are in a low-energy state (high AMP / ATP or ADP / ATP ratio), the α subunit KD and γ subunit of AMPK are tightly cross-linked, and the β subunit is myristoylated, ensuring that the protein phosphatase cannot access the T172 site and remains activated. When cells are in a high-energy state, the KD and γ subunits are loosened, exposing T172 to the phosphatase and causing AMPK inactivation (Steinberg GR, Nature Reviews Drug Discovery. 2019, 18:527).

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

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

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

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

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

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

[0012] Recent studies have shown that NUAK plays a key role in the development of tissue fibrosis through interaction with the transforming growth factor-β (TGF-β) signaling pathway. First, TGF-β upregulates NUAK1 and NUAK2 gene transcription in various epithelial cells, such as keratinocytes and human dermal fibroblasts, while inactivation of MAPK (ERK1 / 2 and p38) signaling inhibits TGF-β-dependent NUAK2 expression. Furthermore, NUAK2 inhibits the intracellular degradation of SMAD3 by binding to the crosslinking domain and MH2 domain of the TGF-β intracellular signaling mediator SMAD3. A similar mechanism also exists between NUAK2 and TβRI. The expression of genes encoding 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, 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 to inhibit fibrosis (van de Vis RAJ, Cancers. 2021, 13: 3377). In general, inhibiting NUAK may effectively inhibit the fibrosis process of participating tissues and organs.

[0013] Because kinases have long been ideal small molecule drug targets due to 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, hypertrophic scars, or simply to alleviate scarring after trauma and surgery). The development of a new generation of NUAK inhibitors has enormous potential for clinical application.

[0014] Summary of the Invention

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

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

[0017] Among them, A is

[0018] X is N or CH, Y is O or NR9;

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

[0020] R3, R4, R7, R 10 、R 11Each is independently halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, or optionally substituted C1-8 alkylamino;

[0021] R6 is an optionally substituted 3-10 membered cycloalkyl, an optionally substituted 3-10 membered heterocycloalkyl, wherein the substituent on the cycloalkyl or heterocycloalkyl is selected from one or more of halogen, amino, hydroxy, nitro, cyano, thiol, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino;

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

[0023] n and r are 0, 1 or 2 respectively, and m, p, q are 0, 1, 2, 3 or 4 respectively.

[0024] In one group of embodiments, Y is O or NH.

[0025] In one group of embodiments, R6 is an optionally substituted 3-10 membered cycloalkyl, preferably an optionally substituted 3-6 membered cycloalkyl, preferably a halogen substituted 3-6 membered cycloalkyl, more preferably R6 is a fluorine substituted 3-6 membered cycloalkyl.

[0026] In one group of embodiments, R6 is

[0027] In one group of embodiments, R5 is Preferably, R5 is

[0028] In one group of embodiments, R5 is substituted at the ortho or meta position relative to Y.

[0029] In one embodiment, the compound represented by formula I is formula I-1,

[0030] In one group of embodiments, X is N and R8 is H or methyl.

[0031] In one group of embodiments, the compound represented by Formula I is Formula I-2 or Formula I-3,

[0032] In one embodiment, the compound represented by formula I is one of the following compounds:

[0033] In one group of embodiments, the pharmaceutically acceptable salt is a formate salt.

[0034] In one set of embodiments, the isotopic derivative is a deuterated derivative.

[0035] On the other hand, the present application also provides a method for preparing the pyrimidineamine compound, which comprises the following steps: preparing a compound of formula I from a compound of formula II and a compound of formula III;

[0036] or

[0037] Prepare a compound of formula V from compounds of formula IV and formula II, and then prepare a compound of formula I from the compound of formula V;

[0038] Wherein, Z is a leaving group, preferably Z is halogen, more preferably Z is Cl, and the definitions of the other groups are as described above.

[0039] In one embodiment, the amino or imino group on the six-membered nitrogen-containing heterocyclic 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.

[0040] In one group of embodiments, the amino or imino group attached to R2 in the compound of formula IV is first protected, and the amino or imino group on the six-membered nitrogen-containing heterocyclic ring in formula II is optionally protected. The protected compound of formula IV is then coupled with the optionally protected compound of formula II, and the protecting group on the amino or imino group attached to R2 is removed to prepare a compound of formula V. The compound of formula V is then reacted with R6-COOH, and the protecting group on ring B is optionally removed to prepare a compound of formula I.

[0041] 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.

[0042] The present application also provides the use of the pyrimidineamine compound for preparing a NUAK1 or NUAK2 inhibitor.

[0043] The present application also 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.

[0044] In one group of embodiments, the disease is Parkinson's disease, Alzheimer's disease, diabetes, hyperlipidemia, obesity, liver cancer, leukemia, lymphoma, tumor metastasis, cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or is used alone to reduce scars after trauma and surgery.

[0045] The beneficial effects of the present invention are:

[0046] 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 or 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

[0047] 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.

[0048] 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.

[0049] 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.

[0050] definition

[0051] Unless otherwise specified, the following terms used in the present invention have the following definitions.

[0052] 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.

[0053] In the present invention, the ring group It means there can be n R3, m R4, p R7, q R 10 、r R 11 Attachment is at any possible position on the ring group.

[0054] In the present invention, some substituents For the A group, it represents the connection site. For the A group, it represents the connection to the amino group at the top and the connection to the N-containing six-membered heterocycle at the bottom.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 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.

[0059] 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.

[0060] The term "halogen" refers to F, Cl, Br, I.

[0061] 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.

[0062] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including configurational isomers and conformational isomers, wherein configurational isomers further include geometric isomers (or cis-trans isomers) and optical isomers (including enantiomers and diastereomers). Geometric isomers may exist in the present compound. Optical isomers refer to substances with identical molecular structures and similar physicochemical properties but different optical rotations. The compounds of the present invention may contain asymmetrically substituted carbon atoms in the R or S configuration, wherein the terms "R" and "S" are defined as in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem. (1976) 45, 13-10. Compounds with asymmetrically substituted carbon atoms (having equal numbers of R and S configurations) are racemic at those carbon atoms. Atoms having an excess of one configuration (relative to the other) result in that configuration being present in a higher number, preferably in an excess of about 85% to 90%, more preferably in an excess of about 9% 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.

[0063] 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.

[0064] The term "isotopic derivative" means that the compounds of the present invention may exist in an isotopically-tagged or enriched form containing one or more atoms having an atomic mass or mass number that is different from the atomic mass or mass number of the atom found in the largest amount in nature. Isotopes may be radioactive or non-radioactive. Isotopes of atoms such as hydrogen, carbon, phosphorus, sulfur, fluorine, chlorine, and iodine include, but are not limited to: 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 32 P, 35 S, 18 F, 36 Cl and 125I. 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.

[0065] The term "hydrate" refers to an association of one or more water molecules with a compound of the present invention.

[0066] The term "solvate" refers to an association between one or more solvent molecules and a compound of the present invention.

[0067] 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] 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.

[0072] Hereinafter, the effects of the specific compounds of the present invention will be described in detail through examples.

[0073] Example

[0074] Example 1 General method for synthesizing compound 518 (TDM-181118)

[0075] Step 1: Compound 518c

[0076] Tert-Butyl (S)-4-(4-((4-(4-(2,2-difluorocyclopropane-1-carboxamido)phenyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate

[0077] 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, 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=0-27 / 73) to give a yellow solid product (compound 518c, 17.3 mg, 49.3% yield). LCMS [M+1] + =540.

[0078] Step 2: Compound 518

[0079] (S)-2,2-Difluoro-N-(4-(2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)cyclopropane-1-carboxamide

[0080] To a solution of compound 518c (17.3 mg, 0.028 mmol) in methanol (0.5 mL) and dichloromethane (2 mL) was added a 4 M hydrochloric acid solution in dioxane (0.07 mL), and the mixture was stirred at room temperature overnight. The reaction was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to give a white solid product (compound 518, TDM-181118, 13.5 mg, 33% yield). LCMS [M+1] + =440.

[0081] 1 H NMR (400MHz, DMSO) δ10.76(s,1H),9.47(s,1H),8.45(d,J=5.1Hz,1H),8.35(s,1H),8.13(d,J=8.7Hz,2H),7.98(s,1H),7.77(d ,J=8.5Hz,2H),7.62(s,1H),7.24(d,J=5.2Hz,1H),4.33(s,1H),3.21(d,J=12.5Hz,2H),2.93-2.75(m,3H),2.13-1.87(m,6H).

[0082] N-(3-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)oxyphenyl)-2,2-difluorocyclopropane-1-carboxamide (TDM-181120) was synthesized by a similar method to Example 1 as a white solid (19.5 mg, yield 13.7%).

[0083] Example 2 General method for the synthesis of compound 527 (TDM-181127)

[0084] Step 1: Compound 527c

[0085] 2,5-Dichloro-4-(3-nitrophenoxy)pyrimidine

[0086] 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), and the mixture was stirred at room temperature for 2 hours. The mixture was then added to water (250 mL) and extracted with ethyl acetate (80 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 slurried with dichloromethane and methanol and filtered to give a white solid product (compound 527c, 1.34 g, 85.9% yield). LCMS [M+1] + =286.

[0087] Step 2: Compound 527d

[0088] 3-((2,5-dichloropyrimidin-4-yl)oxy)aniline

[0089] 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), and the mixture was heated to 65° C. and stirred overnight. The mixture was then filtered and the filtrate was concentrated under reduced pressure to remove tetrahydrofuran. The residue was neutralized with aqueous sodium bicarbonate solution and extracted with ethyl acetate (30 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 (10% methanol in dichloromethane / dichloromethane = 0 to 1 / 99) to give a white solid product (compound 527d, 80 mg, 59.6% yield). LCMS [M+1] + =256.

[0090] Step 3: Compound 527f

[0091] N-(3-((2,5-dichloropyrimidin-4-yl)oxy)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0092] To a solution of compound 527d (80 mg, 0.312 mmol), compound 527e (57 mg, 0.469 mmol) and 2-(7-azabenzotriazole)-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), and the mixture was heated to 50°C and stirred for 2 hours. The mixture was added with water (80 mL), then extracted with ethyl acetate (50 mL*3), the organic layers were combined, washed with brine, dried over sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (10% methanol in dichloromethane / dichloromethane = 0 to 1 / 99) to give a yellow solid product (compound 527f, 70 mg, 62.3% yield). LCMS [M+1] + =360.

[0093] Step 4: Compound 527h

[0094] 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

[0095] 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 mmmol), 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, 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 = 0-30 / 70) to give a yellow solid product (compound 527h, 55 mg, 48% yield). LCMS [M+1] + =590.

[0096] Step 5: Compound 527

[0097] N-(3-((5-chloro-2-((1-(piperidin-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)oxy)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0098] To a solution of compound 527h (55 mg, 0.093 mmol) in methanol (1 mL) and dichloromethane (5 mL) was added a dioxane hydrochloride solution (0.35 mL), and the mixture was stirred at room temperature overnight. The mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (formic acid) to give a white solid product (compound 527, TDM-181127, 25 mg, 54.9% yield). LCMS [M+1] + =490.

[0099] 1 H NMR(400MHz,MeOD)δ8.55(s,1H),8.26(s,1H),7.99(s,1H),7.53(s,1H),7.23(d,J=30.0Hz,2H),7.10–6.81(m,2H),4 .01(s,1H),3.52-3.40(m,2H),3.15(dd,J=20.8,7.9Hz,2H),2.71(ddd,J=13.2,10.8,7.8Hz,1H),2.20-1.81(m,6H).

[0100] Example 3 General method for the synthesis of compound 530 (TDM-181130)

[0101] Step 1: Compound 530c

[0102] 2,5-Dichloro-N-methyl-N-(3-nitrophenyl)pyrimidin-4-amine

[0103] To a solution of compound 530b (730 mg, 4.8 mmol) in N,N-dimethylformamide (40 mL) at 0°C was added sodium hydroxide (384 mg, 9.6 mmol). The mixture was naturally warmed to room temperature and stirred for one hour. Compound 530a (1232 mg, 6.72 mmol) was then added and stirred at room temperature for two hours. The reaction was detected to be complete. Post-treatment: The reaction solution was poured into water, and the aqueous phase was extracted three times with ethyl acetate (3 x 150 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: EA / PE = 0-10%) to afford the title compound (compound 530c, 708.6 mg, 49.35% yield) as a yellow solid. LCMS [M+1] + =299,301.

[0104] Step 2: Compound 530d

[0105] N1-(2,5-Dichloropyrimidin-4-yl)-N1-methylbenzene-1,3-diamine

[0106] 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 detected to be complete. Post-treatment: The reaction mixture was cooled to room temperature and filtered. The filter cake was washed with ethyl acetate. The filtrate was extracted three times with ethyl acetate. 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 obtain the title compound (compound 530d, 603 mg, 95% yield) as a yellow solid. LCMS [M+1] + =269,271.

[0107] Step 3: Compound 530f

[0108] (S)-N-(3-((2,5-dichloropyrimidin-4-yl)(methyl)amino)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0109] To a solution of compound 530d (537.4 mg, 2 mmol) in pyridine (20 mL) was added compound 530e (244 mg, 2 mmol). The reaction solution was cooled to 0°C, and phosphorus oxychloride (0.28 mL, 3 mmol) was added. The mixture was stirred at this temperature for one hour until the reaction was complete. Post-treatment: The reaction solution was poured into ice water (30 mL). The aqueous phase was extracted twice with ethyl acetate (2 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography using an eluent of EA / PE = 0-40% to afford the title compound (compound 530f, 273.5 mg, 36.6% yield) as a yellow solid. LCMS [M+1] + =373,375.

[0110] Step 4: Compound 530

[0111] (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

[0112] To a solution of compound 530f (52 mg, 0.14 mmol) in n-butanol (5 mL) were 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 hours. The reaction was detected to be complete. Post-treatment: The reaction mixture was poured into water, and the aqueous phase was extracted twice with ethyl acetate (2 x 150 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled to dryness. The crude product was purified to obtain the title compound (compound 530, 15.4 mg, 19.7% yield) as a yellow solid. LCMS [M+1] + =558.

[0113] 1 H NMR (400MHz, DMSO) δ10.46(s,1H),7.98(s,1H),7.87(s,1H),7.73(d,J=8.7Hz,1H),7 .40(dd,J=8.0,5.0Hz,2H),7.31(t,J=8.0Hz,1H),6.91(d,J=9.1Hz,1H),6.63(d,J=2. 5Hz,1H),6.47(dd,J=8.8,2.5Hz,1H),3.83(s,3H),3.36(s,3H),3.15-3.07(m,4H),2. 77(ddd,J=13.6,10.9,8.1Hz,1H),2.47-2.43(m,4H),2.22(s,3H),2.06-1.90(m,2H).

[0114] (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 by a method similar to Example 3 as an off-white solid (9.4 mg, yield 10%).

[0115] Example 4 General method for the synthesis of compound 532 (TDM-181132)

[0116] Step 1: Compound 532c

[0117] 2,5-Dichloro-4-(2-nitrophenoxy)pyrimidine

[0118] To a solution of compound 532b (421 mg, 3.03 mmol) in N,N-dimethylformamide (20 mL) were 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 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: PE / EA = 0-15%) to obtain the title compound (compound 532c, 800 mg, 92.4% yield) as a white solid. LCMS [M+1] + =286,288.

[0119] Step 2: Compound 532d

[0120] 2-((2,5-dichloropyrimidin-4-yl)oxy)aniline

[0121] To a solution of compound 532c (1.54 g, 5.4 mmol) in tetrahydrofuran (150 mL) and water (150 mL) were 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. The reaction was detected to be complete. Post-treatment: The reaction mixture was cooled to room temperature and filtered. The filtrate was drained and extracted three times with water and ethyl acetate (20 mL). 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 using a solvent of (dichloromethane / methanol = 10:1) / DCM = 0-40% to obtain the title compound (compound 532d, 476 mg, 34% yield) as a yellow solid. LCMS [M+1] + =256,258.

[0122] Step 3: Compound 532f

[0123] N-(2-((2,5-Dichloropyrimidin-4-yl)oxy)phenyl)cyclopropanecarboxamide

[0124] 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). The reaction solution was cooled to 0°C and phosphorus oxychloride (0.22 mL, 2.39 mmol) was added. The mixture was stirred at this temperature for one hour until the reaction was complete. Post-treatment: The reaction solution was poured into ice water (30 mL). The aqueous phase was extracted twice with ethyl acetate (2 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography using an eluent of EA / PE = 0-40% to obtain the title compound (compound 532f, 396.8 mg, 59% yield) as a yellow solid. LCMS [M+1] + =325.

[0125] Step 4: Compound 532

[0126] N-(2-(5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)oxy)phenyl)cyclopropanecarboxamide

[0127] 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 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 two hours until the reaction was complete. Post-treatment: The reaction solution was poured into ice water (30 mL), and the aqueous phase was extracted twice with ethyl acetate (2 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled to dryness. The crude product obtained afforded the title compound (compound 532, 14.3 mg, 19.7% yield) as a yellow solid. LCMS [M+1] + =255,509.

[0128] 1 H NMR (400MHz, DMSO) δ8.30(s,1H),8.07(s,1H),7.72-7.64(m,1H),7.56(s,2H),7.23(d,J=5.9Hz,3H),6.58(d,J=2.2Hz,1 H), 6.32 (d, J = 8.4Hz, 1H), 3.77 (s, 3H), 3.08 (s, 4H), 2.45 (s, 4H), 2.22 (s, 3H), 1.76 (d, J = 5.0Hz, 1H), 1.00-0.86 (m, 4H).

[0129] Example 5 General method for the synthesis of compound 560 (TDM-181160)

[0130] Step 1: Compound 560c

[0131] 2,5-Dichloro-4-(2-nitrophenoxy)pyrimidine

[0132] To a solution of compound 560b (421 mg, 3.03 mmol) in N,N-dimethylformamide (20 mL) were 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 determined to be complete. Post-treatment: The reaction mixture was poured into water, and ethyl acetate (3 x 50 mL) was added. The mixture was extracted three times in separate layers. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography using an eluent of PE / EA = 0-15% to obtain the title compound (compound 560c, 800 mg, 92.4% yield) as a white solid. LCMS [M+1] + =286,288.

[0133] Step 2: Compound 560d

[0134] 2-((2,5-dichloropyrimidin-4-yl)oxy)aniline

[0135] To a solution of compound 560c (1.54 g, 5.4 mmol) in tetrahydrofuran (150 mL) and water (150 mL) were added iron powder (1.51 g, 27 mmol) and ammonium chloride (1.44 g, 27 mmol). The reaction solution was heated to 65°C and stirred for 3 hours. The reaction was detected to be complete. Post-treatment: The reaction solution was cooled to room temperature and filtered. The filter cake was washed with ethyl acetate. The filtrate was extracted three times with ethyl acetate. 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 using an eluent of PE / EA = 0-50% to obtain the title compound (compound 560d, 476 mg, 34% yield) as a yellow solid. LCMS [M+1] + =256,258.

[0136] Step 3: Compound 560f

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

[0138] 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). The reaction solution was cooled to 0°C and phosphorus oxychloride (0.22 mL, 2.39 mmol) was added. The reaction was incubated for 30 minutes until completion was detected. Post-treatment: The reaction solution was poured into ice water (30 mL). The aqueous phase was extracted twice 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 using an eluent of PE / EA = 0-20% to afford the title compound (compound 560f, 396.8 mg, 59% yield) as a yellow solid. LCMS [M+1] + =360,362.

[0139] Step 4: Compound 560

[0140] (S)-N-(2-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)oxy)benzene)-2,2-difluorocyclopropane-1-carboxamide

[0141] 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 detected to be complete. Post-treatment: The reaction mixture was poured into water, and the aqueous phase was extracted with ethyl acetate (2 x 150 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 using dichloromethane:methanol = 10:1 / DCM = 0-50%. The resulting crude product was purified to obtain the title compound (compound 560, 12.7 mg, 17% yield) as a white solid. LCMS [M+1] + =545.2.

[0142] 1H NMR (400MHz, DMSO) δ8.38 (s, 1H), 8.06 (s, 1H), 7.70 (dd, J = 7.0, 3.0Hz, 1H), 7.5 5(d,J=12.4Hz,2H),7.26(dd,J=6.3,3.7Hz,3H),6.58(d,J=2.5Hz,1H),6.31(d ,J=8.7Hz,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, 5.8Hz, 1H), 2.02 (td, J = 14.1, 7.9Hz, 1H).

[0143] Example 6 General method for the synthesis of compound 568 (TDM-181168)

[0144] Step 1: Compound 568b

[0145] N1-(2,5-Dichloropyrimidin-4-yl)-N1-methylbenzene-1,2-diamine

[0146] To a solution of compound 568a (243 mg, 0.813 mmol) in tetrahydrofuran (2.5 mL) and water (2.5 mL) were added iron powder (28 mg, 0.502 mmol) and ammonium chloride (26.8 mg, 0.502 mmol), and 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 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 (100% dichloromethane) to give a yellow solid product (compound 568b, 112.3 mg, 45.7% yield). LCMS [M+1] + =269,271.

[0147] Step 2: Compound 568d

[0148] (S)-N-(2-((2,5-dichloropyrimidin-4-yl)(methyl)amino)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0149] To a solution of compound 568b (112.3 mg, 0.417 mmol) and compound 568c (61 mg, 0.501 mmol) in pyridine (5 mL) was added phosphorus oxychloride (96 mg, 0.626 mmol) at 0°C under an argon atmosphere, and 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.

[0150] Step 3: Compound 568

[0151] (S)-N-(2-((5-chloro-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)(methyl)amino)phenyl)-2,2-difluorocyclopropane-1-carboxamide

[0152] 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) was added p-toluenesulfonic acid monohydrate (71.5 mg, 0.376 mmol), and the mixture was heated to 110° C. and stirred for 6 hours. The mixture was added with water (50 mL) and extracted with ethyl acetate (25 mL*3). The organic layers were combined and washed with brine. The filtrate was concentrated under reduced pressure, and 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.

[0153] 1 H NMR (400MHz, DMSO) δ9.86(s,1H),9.23(s,1H),8.16(s,1H),8.05(d,J=8.2Hz,1H),7.93(s,1H),7.59(d,J=9.0Hz,2 H),7.32-7.23(m,1H),7.09(d,J=4.1Hz,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).

[0154] Test example: Enzyme activity inhibition assay of NUAK1 / NUAK2 kinase inhibitors

[0155] 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:

[0156] 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.

[0157] Test steps:

[0158] 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).

[0159] 2. Add kinase and mix gently (the final concentration of NUAK1 is 10 nM; the final concentration of NUAK2 is 50 nM);

[0160] 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;

[0161] 4. Join 33 P-labeled ATP (unlabeled ATP / labeled ATP ratio of 25:1 or 2.5:1);

[0162] 5. Incubate at room temperature for 2 hours;

[0163] 6. Detect kinase activity using P81 filter paper combined with hotspot technology.

[0164] The IC of the test compound against NUAK1 / NUAK2 was calculated according to the above assay. 50 , see Table 1 for specific results.

[0165] IC 50 Calculations were made using the formula derived from a sigmoidal dose-response curve (variable slope):

[0166] 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.

[0167] Table 1 IC values ​​of the test compounds of this application for NUAK1 / NUAK2 50 (nM)

[0168] 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 The concentration can reach several nM or tens of nM. Therefore, the above experiments have demonstrated that the pyrimidineamine compounds of the present application can be used as NUAK1 / NUAK2 inhibitors. The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, various simple variations of the technical solution of the present invention can be made, and these simple variations all fall within the scope of protection of the present invention.

[0169] 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.

[0170] In addition, the various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. A pyrimidine amine compound, characterized in that The pyrimidineamine compound is a compound represented by formula I, or a stereoisomer, tautomer, isotope derivative, hydrate, solvate, prodrug, and pharmaceutically acceptable salt thereof; Among them, A is X is N or CH, Y is O or NR9; R1, R2, R8, and R9 are each independently H, or an optionally substituted C1-8 alkyl group; R3, R4, R7, R 10 , R 11 Each is independently halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino; R6 is an optionally substituted 3-10 membered cycloalkyl, an optionally substituted 3-10 membered heterocycloalkyl, wherein the substituent on the cycloalkyl or heterocycloalkyl is selected from one or more of halogen, amino, hydroxy, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkyloxy, optionally substituted C1-8 alkylthio, and optionally substituted C1-8 alkylamino; The substituent on the C1-8 alkyl group is selected from one or more of halogen, amino, hydroxyl, nitro, cyano, and mercapto; n and r are 0, 1 or 2 respectively, and m, p, q are 0, 1, 2, 3 or 4 respectively.

2. The compound according to claim 1, characterized in that Y is O or NH.

3. The compound according to any one of claims 1-2, characterized in that R6 is an optionally substituted 3-10 membered cycloalkyl group.

4. The compound according to claim 3, characterized in that R6 is an optionally substituted 3-6 membered cycloalkyl group.

5. The compound according to claim 4, characterized in that R6 is a 3-6 membered cycloalkyl group substituted with halogen.

6. The compound according to claim 5, characterized in that R6 is a fluorine-substituted 3-6-membered cycloalkyl group.

7. The compound according to claim 4, characterized in that R6 is 8. The compound according to claim 7, characterized in that R5 is 9. The compound according to claim 8, characterized in that R5 is 10. The compound according to any one of claims 1 to 9, characterized in that R5 is substituted at the ortho or meta position of Y.

11. The compound according to any one of claims 1 to 10, characterized in that The compound represented by formula I is 12. The compound according to claim 11, characterized in that X is N, and R8 is H or methyl.

13. The compound according to claim 12, characterized in that The compound represented by formula I is 14. The compound according to claims 1-13, characterized in that The compound represented by formula I is one of the following compounds:

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

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

17. A method for preparing a compound according to any one of claims 1 to 16, comprising the following steps: The compound of formula I is prepared from the compound of formula II and the compound of formula III; or The compound of formula V is prepared from the compound of formula IV and formula II, and the compound of formula I is prepared from the compound of formula V; Optionally, the preparation process includes necessary protection and deprotection steps; in, Z is a leaving group, preferably Z is halogen, more preferably Z is Cl, and the definitions of other groups are the same as those in claims 1-16.

18. A pharmaceutical composition, characterized in that The compound described in any one of claims 1 to 16 is used as an active ingredient.

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

20. Use of the compound according to any one of claims 1 to 16 for preparing a NUAK1 or NUAK2 inhibitor.

21. Use of the compound according to any one of claims 1 to 16 for preparing a medicament, characterized in that The compound is used for preventing or treating the following diseases by inhibiting NUAK1 or NUAK2: neuropsychiatric diseases, metabolic diseases, tumors, visceral fibrosis diseases, and skin fibrosis diseases.

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, cirrhosis, renal fibrosis, pulmonary fibrosis, sequelae of myocarditis, scleroderma, keloid, hypertrophic scar, or is used alone to reduce trauma and postoperative scars.

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