Kinase inhibitors

Abstract

This invention discloses compounds that inhibit kinases Lck or Btk, including pharmaceutically acceptable salts, hydrides, stereoisomers, and pharmaceutical compositions thereof.

Description

Background of the Invention

[0002] Autoimmune diseases, also known as autoimmune disorders, are characterized by an overactive immune response or an incorrect targeting of the body's own tissues. 1 Traditionally, autoimmune diseases were thought to be caused by damage mediated by T cells and / or autoantibodies. However, recent research suggests that T and B lymphocytes, as well as innate immunity, play important roles in the pathogenesis of these diseases. 2 At the molecular level, activation of T and B lymphocytes depends on a signaling cascade via immune receptors, such as the T-cell antigen receptor (TCR) and B-cell antigen receptor (BCR). These immune receptors are activated by ligand binding, which in turn leads to the activation of downstream kinase targets. A complex network of protein tyrosine kinases (PTKs) is associated with important pathways. Activated PTKs then phosphorylate receptor-associated transmembrane adaptors (ITAMs) on their cytoplasmic surface. Through a series of post-translational modifications, such as phosphorylation, downstream effectors, such as transcription factors that regulate the expression of various inflammatory cytokines, are initiated. 3,4 .

[0003] Protein kinases play a crucial role in the activation of B and T cells. As responses to changes in extracellular mediators and the environment, protein kinases participate in signaling events controlling T and B cell activation, growth, and differentiation. Some small molecule kinase inhibitors have shown potential for treating diseases such as rheumatoid arthritis and psoriasis. 5-9 .

[0004] In overactive T lymphocytes, TCR-mediated signaling pathways lead to the upregulation of inflammatory cytokines, such as IL-2 and interferon-γ (IFN-γ), which stimulate immune responses. It has been established that a key initiating event for T cell activation is the increased phosphorylation of the immune receptor tyrosine-based activation motif (ITAM) in the TCR-CD3 complex, a process mediated by Src family PTKs. The Src family consists of eight distinct members: Src, Lck, Fyn, Lyn, Hck, Fgr, Blk, and Yes. Among them, lymphocyte-specific protein kinase (Lck) and proto-oncogene tyrosine protein kinase (Fyn) are expressed within T cells and are located at the initiation stage of the TCR-mediated signaling cascade. 3,10,11 .

[0005] Lck is primarily expressed in T cells and its composition is associated with the cytoplasmic portions of CD4 and CD8 surface receptors. In mature peripheral T cells, Lck phosphorylates the TCR and initiates TCR-linked signal transduction. Lck is also expressed at all stages of thymocyte development and plays a crucial role in the selection and maturation of developing T cells. 10,11 .

[0006] Specifically, Lck plays a role in phosphorylating ITAM during the initiation of TCR-mediated signaling pathways, providing a binding site for the tandem SH2 domain of the Syk family kinase ZAP-70. Once recruited, ZAP-70 is phosphorylated and activated by Lck at Tyr493 of its activation loop. 12 This leads to the autophosphorylation of ZAP-70. Activated ZAP-70 then phosphorylates adaptor proteins, such as LAT, which act as a scaffold to recruit downstream signaling molecules. This cascade ends by initiating transcription of genes associated with cytokine release (especially IL-2), ultimately promoting T cell proliferation. 13 Interference with LCK kinase activity can produce strong immunosuppressive effects. For example, imatinib reduces TCR-induced proliferation and activation by inhibiting LCK. 14 Orally administered selective Lck inhibitor A-770041 (IC50) 50 =147 nM) can inhibit the production of IL-2 in whole blood stimulated by concanavalin A, and the EC50 of this effect is [missing information]. 50 Approximately 80 nM 15 At a dose of 10 mg / kg / day, A-770041 showed that it prevented rejection of heterotopic transplanted hearts across the major histocompatibility barrier for at least 65 days. 15,16 Rosmarinic acid (RosA), which inhibits LCK by targeting the SH2 domain, also showed inhibitory effects on T cell activation and proliferation. 17 When administered daily at a dose of 50 mg / kg / day for 15 consecutive days, RosA inhibited synovitis in a mouse model of collagen-induced arthritis. 18 The methyl ester derivative (RosA-Me) exhibits a significantly stronger inhibitory effect on IL-2 gene expression and T cell proliferation than its counterpart, RosA. Compared to RosA and MTX, oral administration of RosA-Me at a dose of 50 mg / kg / day to mice significantly reduced inflammation and arthritis markers. 19 .

[0007] As another member of the Src family, Fyn's function partially overlaps with Lck's in initiating tyrosine phosphorylation of the TCR. Both kinases interact with the TCR and increase the production of interleukin-2 (IL-2). 20,21 Genetic evidence demonstrates that Fyn activation is directly associated with TCR-induced Lck translocation, suggesting that Fyn participates in T cell activation. However, studies in Lck-deficient and Fyn-deficient mice have shown that these kinases play a limited role in development. 22-24 Studies on the Lck-deficient T cell line (JCaM1) have confirmed that the TCR can still be phosphorylated at certain tyrosine residues, which promotes the recruitment of ZAP-70 kinase. In contrast, the phosphorylation pattern of the TCR is altered, and ZAP-70 activation is defective. During this process, the expression level of the molecular marker CD69 is increased, while NFAT activation and interleukin-2 production are significantly reduced. These results indicate that Fyn can catalyze the phosphorylation of ITAM in the absence of Lck, but cannot largely compensate for the lack of Lck. Furthermore, the outcome of TCR signal transduction can be determined based on the choice of which member of the Src family of kinases initiates the signaling cascade. 25 .

[0008] The dual fatty acylation process of myristylation and palmitylation is crucial in the initiation of the TCR signaling pathway by Fyn and Lck. 2-Bromopalmitate can effectively block palmitoylation of Fyn. In Jurkat T cells, 2-Bromopalmitate blocks the localization of endogenously palmitoylated Fyn and Lck on the detergent membrane, thereby inhibiting the TCR signaling pathway and T cell activation. 26 Furthermore, it was found that unsaturated fatty acids (PUFAs), especially the n-3 series, can also inhibit the fatty acylation of Fyn. 26,27 Glucocorticoids (GCs) are used clinically as immunosuppressants. GCs are potent immunosuppressants that rapidly inhibit the recruitment of Fyn and Lck to the T-cell receptor complex. These results clarify Lck and Fyn kinases as molecular targets of GC mediated through the GC receptor-dependent pathway. 28 Simultaneous inhibition of Fyn and Lck can suppress autoimmune diseases.

[0009] The success of B-cell depletion therapy (BCDT), such as the clinical application of rituximab (MabThera / Rituxan; Biogen Idec / Genentech) in the treatment of rheumatoid arthritis, suggests the important role of B cells in certain autoimmune diseases. 29Tec family kinase BTK is expressed only in B cells, monocytes, macrophages, neutrophils, and mast cells; it has not been found to be expressed in T cells or natural killer (NK) cells. BTK is a key regulator of B cell development, activation, signaling, and survival. BTK is particularly important for B cell activation following the involvement of the B cell receptor (BCR). 30 Upon stimulation by BCR, BTK is activated by upstream Src family kinases Blk, Lyn, and Fyn. Activated BTK then phosphorylates and activates phospholipase Cγ2 (PLCγ2), which catalyzes the production of DAG and IP3, leading to stimulation of downstream signaling molecules such as transcription factors NF-κB and NFAT. Functional mutations in Btk induce X-linked immunodeficiency (XID) in mice, characterized by decreased serum Ig levels. 31 Btk functional nonsense mutations in humans lead to primary immunodeficiency diseases characterized by peripheral B cell deficiency and extremely low serum immunoglobulin (Ig) levels.

[0010] Some covalently bound compounds targeting specific residues (Cys481) in Btk, such as ibrutinib (PCI-32765) and AVL-292, have shown IC50 efficacy in established CIA mouse models. 50 High binding affinity below 0.5 nM and oral efficacy mean dose-dependent inhibition of clinical arthritis scores and anti-collagen autoantibody production. 6 Furthermore, in the MRL-Fas(lpr) lupus model, PCI-32765 also inhibited autoantibody production and the progression of kidney disease. 6 Another Btk inhibitor, RN486 (IC), 50 =4.0 nM) can completely halt disease progression in the NZB×NZW systemic lupus erythematosus (SLE) mouse model. RN486 administration significantly inhibits inflammatory responses in PCA (type I hypersensitivity) or rPCA (type III hypersensitivity) mouse models. In RA models, oral administration of RN486 reduces paw swelling and inflammatory markers by suppressing joint and systemic inflammation. 32 .

[0011] In addition to the kinases involved in the proximal signaling pathways of the TCR and BCR mentioned above, cumulative studies have suggested that p38MAPK plays an important role in the pathogenesis of several immune-mediated diseases, including rheumatoid arthritis (RA), Sjögren's syndrome, systemic lupus erythematosus (SLE), inflammatory bowel disease (IBD), and psoriasis. In the TCR and BCR signaling pathways, the MAPK cascade is activated by Vav. As a downstream effector, Rac1 acts as a MAPKK kinase (MAPKKK) and is located upstream of p38 MAPK. Upon activation, p38 MAPK regulates the expression of tumor necrosis factor (TNF)α, interferon (IFN)γ, and other cytokines such as IL-1, TL-6, and IL-17 through transcriptional and post-transcriptional mechanisms. 33,34 The success of antibody therapy suggests the crucial role of cytokines and p38 MAPK in the development of autoimmune diseases. Several small p38 inhibitors have shown promising results in preclinical and early clinical studies. However, most subsequent clinical trials have failed, either due to poor efficacy or severe toxicity. The ATP-occupying p38 MAPK inhibitor VX-745, despite promising results in a phase II clinical trial for RA, was discontinued due to hepatotoxicity. 35 In a DSS-induced colitis model, SB203580 treatment improved clinical scores by reducing the mRNA levels of pro-inflammatory cytokines in vivo. 36 In addition, some highly selective non-ATP competitive inhibitors, such as VX-702, SCIO469, and BIRB796, have also shown transient inhibitory effects on pro-inflammatory disease markers and have not been able to completely control disease exacerbation or progression. 37 .

[0012] Unacceptable toxicity and low efficacy in clinical trials suggest that p38 MAPK inhibitor administration may not be the best solution for anti-inflammatory treatment. Further research indicates that p38 MAPK plays a multifaceted role in regulating many cellular processes beyond inflammation. This functional diversity is the reason for the high toxicity and strong inhibitory effects. Moreover, the existence of compensatory mechanisms appears to further weaken the effectiveness of treatment. These studies all highlight the need to develop novel agents that simultaneously target more pathologically relevant upstream kinases. Inhibitors with multi-targeting properties and moderate but balanced affinity represent a better treatment option in terms of limited toxicity and sustained clinical response. Invention Overview

[0014] This invention provides compounds of formula I:

[0015] in:

[0016] R1–R5 are each independently H, halogen, hydroxyl, methyl, trifluoromethyl, or methoxy, especially wherein R1 and R5 are each independently halogen, hydroxyl, methyl, or trifluoromethyl; and

[0017] R6 is an optionally substituted Cn cyclic hydrocarbon group, wherein n = 3-18, and contains at most n-1 heteroatoms, each independently selected from N, O, S and P, in particular, wherein the hydrocarbon group is selected from cycloalkyl, aryl, heteroaryl or heterocyclic groups;

[0018] Or its stereoisomers or pharmaceutically acceptable salts.

[0019] In a specific implementation:

[0020] –n is 3, 4, 5, 6, 8, 9, or 10; or n is 3, 5, 6, or 9;

[0021] -R6 is a substituted or unsubstituted 5- or 6-membered allotropic ring or heterocyclic ring, or a 9- or 10-membered bicyclic aryl group;

[0022] -R6 is optionally substituted with: cyclopropyl; a 5-membered aryl group selected from pyrrole, azoles (e.g., pyrazole, imidazole, triazole, tetraazole, pentaazole, oxazole, isoxazole, thiazole, or isothiazole), furan, m-dioxacyclopentenethiophene, dithiacyclopentadiene, or oxothiacyclopentadiene and their reduced forms (e.g., dihydrofuran, dihydroimidazole); preferably a 2-substituted moiety such as 2-azole, 2-pyrrole, 2-azole (e.g., 2-pyrazole, 2-imidazole, 2-oxazole, 2-isooxazole, 2-thiazole, or 2-isothiaazole), 2-furan, 2-thiophene, 2-oxacyclopentadiene, m-dioxacyclopentadiene, or 2-thiacyclopentadiene (2-thiophene); a 6-membered aryl group selected from phenyl and pyridine; or a 9-membered aryl group is benzimidazole;

[0023] -R6 represents phenyl, 3-substituted phenyl, 3,4-substituted phenyl, or 3,4,5-substituted phenyl;

[0024] The substituents of -R6 are selected from: halogens, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR"CO2R', -NR'-C(O)NR"R'", -NR'-SO2NR"R'", -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR R', -S(O)R', -SO2R', -SO2NR'R", -NR"SO2R, -N3, -CH(Ph)2, perfluoro(C1-C4)alkoxy and perfluoro(C1-C4)alkyl, the number of substituents being 0 to the total number of open valences on the aromatic ring system; wherein R', R" and R'" are each independently selected from hydrogen, (C1-C8)alkyl and heteroalkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C1-C4)alkyl and (unsubstituted aryl)oxy-(C1-C4)alkyl;

[0025] -R2-R4 are H, and at least one of R1 and R5 is halogen, hydroxyl, methyl, trifluoromethyl or methoxy;

[0026] -R2-R4 is H, and R1 and R5 are halogens, hydroxyl groups, methyl groups, trifluoromethyl groups, or methoxy groups;

[0027] – All its combinations, such as, where:

[0028] R2-R4 are H, and R1 and R5 are halogens, hydroxyl groups, methyl groups, trifluoromethyl groups, or methoxy groups;

[0029] R6 is a substituted or unsubstituted 5- or 6-membered allotropic ring or heterocyclic ring, or a 9- or 10-membered bicyclic aryl group; and

[0030] The substituents of R6 are selected from: halogens, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR"CO2R', -NR'-C(O)NR"R'", -NR'-SO2NR"R'", -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -SO2R', -SO2NR'R", -NR"SO2R, -N3, -CH(Ph)2, perfluoro(C1-C4)alkoxy and perfluoro(C1-C4)alkyl, with the number of substituents ranging from 0 to the total number of open valences on the aromatic ring system; wherein R', R" and R'" are each independently selected from hydrogen, (C1-C8)alkyl and heteroalkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C1-C4)alkyl and (unsubstituted aryl)oxy-(C1-C4)alkyl; and / or

[0031] These compounds exhibited an IC50-like activity in kinase activity assays. 50 It exhibits inhibitory activity against Lck or Btk at 10 μM or less.

[0032] Representative and exemplary compounds include the following structures, or their stereoisomers, or their pharmaceutically acceptable salts:

[0033]

[0034]

[0035]

[0036]

[0037] This invention provides pharmaceutical compositions comprising a subject compound. In some aspects, the invention provides pharmaceutical compositions comprising a therapeutically effective amount of a subject compound in a unit dose form and one or more pharmaceutically acceptable excipients. In some aspects, the invention provides compositions comprising a therapeutically effective amount of a subject compound and various therapeutic agents having therapeutic activity against autoimmune and / or inflammatory diseases or cancer.

[0038] The present invention also provides methods for manufacturing and using the subject compound, including methods for inhibiting kinase activity. In some aspects, the present invention provides methods for treating diseases associated with undesirable kinase activity, comprising administering an effective amount of the subject compound or a prodrug thereof to a person in need, wherein the disease is an allergic disease, an autoimmune disease, an inflammatory disease, or cancer, wherein the method may further include a preliminary step of diagnosing the disease or cancer, or a subsequent step of detecting improvement in the disease or cancer.

[0039] This invention includes all combinations of the specific embodiments described herein.

[0040] Detailed Description of the Invention

[0041] Specific embodiments and examples are described below in an illustrative rather than limiting manner. Those skilled in the art will readily recognize that many non-critical parameters can be modified or altered to produce substantially similar results. Numerous embodiments are provided in this invention.

[0042] This invention encompasses all possible combinations, as each is explicitly enumerated; thus, aspects and embodiments of the invention include, for example, combinations in which R1 is a substituted or unsubstituted phenyl; R2 is H, hydroxyl, C1-C4 alkyl or C1-C4 alkoxy, R3 is H or methyl, and R4 is 1-dimethylpropyl.

[0043] Unless it is found to be improper or otherwise noted, in these descriptions and throughout the specification, indefinite articles indicate one or more, the term "or" indicates and / or, and polynucleotide sequences should be understood to include the reverse strand and other alternative backbones described herein. Furthermore, this specification uses abbreviations to describe classes to enumerate all members of that class; for example, a description abbreviated as (C1-C3)alkyl indicates that all C1-C3 alkyl groups are enumerated: methyl, ethyl, and propyl, including their isomers.

[0044] Unless otherwise stated in the context in which they are applied, the following words, phrases and symbols are generally intended to convey the following meanings.

[0045] As used in this invention, the term "heteroatom" generally refers to any atom other than carbon or hydrogen. Preferred heteroatoms include oxygen (O), phosphorus (P), sulfur (S), nitrogen (N), and halogens. Preferred heteroatom functional groups are haloformyl, hydroxyl, aldehyde, amino, azo, carboxyl, cyano, thiocyanyl, carbonyl, halogen, hydroperoxyl, imino, aldehyde imino, isocyanide, isocyanate, nitrate, nitrile, nitrite, nitro, nitrosyl, phosphate, phosphonyl, sulfur, sulfonyl, sulfonyl, and mercapto.

[0046] Unless otherwise stated, the term "alkyl" itself, or as part of another substituent, refers to a fully saturated, straight-chain or branched, or cyclic hydrocarbon group, containing a specified number of carbon atoms (i.e., C1-C8 represents 1-8 carbon atoms), or a combination thereof. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, homologues and isomers, such as homologues and isomers of n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0047] The term "alkenyl" itself, or as part of another substituent, refers to a straight-chain or branched, or cyclic, hydrocarbon group, or a combination thereof, that is monounsaturated or polyunsaturated and contains a specified number of carbon atoms (i.e., C2-C8 represents 2-8 carbon atoms) and one or more double bonds. Examples of alkenyl groups include vinyl, 2-propenyl, crotonyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), and their higher homologues and isomers.

[0048] The term "alkynyl" itself, or as part of another substituent, refers to a straight-chain or branched hydrocarbon group, or a combination thereof, that is monounsaturated or polyunsaturated, containing a specified number of carbon atoms (i.e., C2-C8 represents 2-8 carbon atoms) and one or more triple bonds. Examples of alkynyl groups include ethynyl, 1-propynyl and 3-propynyl, 3-butynyl, and their higher homologues and isomers.

[0049] The term "alkylene" itself, or as part of another substituent, refers to a divalent group derived from an alkyl group, exemplified by -CH2-CH2-CH2-CH2-. Typically, alkyl (or alkylene) groups have 1-24 carbon atoms, with groups having 10 or fewer carbon atoms being preferred in this invention. "Lower alkyl" or "lower alkylene" refers to a shorter-chain alkyl or alkylene group that typically has 8 or fewer carbon atoms.

[0050] The terms “alkoxy,” “alkylamine,” and “alkylthio” (or thioalkoxy) are used in their conventional sense to refer to those alkyl groups that are attached to the remainder of the molecule by an oxygen atom, an amino atom, or a sulfur atom, respectively.

[0051] Unless otherwise stated, the term "heteroalkyl" on its own or in combination with another term means a stable straight-chain or branched, or cyclic, hydrocarbon group, or a combination thereof, consisting of the stated number of carbon atoms and one to three heteroatoms selected from O, N, P, Si, and S, wherein nitrogen, sulfur, and phosphorus atoms may optionally be oxidized, and nitrogen heteroatoms may optionally be quaternized. Heteroatoms O, N, P, and S may be located at any position within the heteroalkyl group. Heteroatoms Si may be located at any position within the heteroalkyl group, including positions where the alkyl group is attached to the remainder of the molecule. Examples include -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, and -CH=CH-N(CH3)2. Up to two heteroatoms can be consecutive, such as, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0052] Similarly, the term "heteroalkylene" itself, or as part of another substituent, indicates a divalent group derived from a heteroalkyl group, exemplified by -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-. For heteroalkylene, the heteroatom can also occupy one or both chain ends (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Furthermore, the orientation of the linking group for alkylene and heteroalkylene is not implied.

[0053] Unless otherwise stated, the terms "cycloalkyl" and "heterocycloalkyl," whether used alone or in combination with other terms, refer to "alkyl" and "heteroalkyl" in cyclic form, respectively. Thus, a cycloalkyl group has a specified number of carbon atoms (i.e., C3-C8 represents 3-8 carbons) and may also contain one or two double bonds. A heterocycloalkyl group has a specified number of carbon atoms and 1-3 heteroatoms selected from O, N, Si, and S, wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. Furthermore, in the case of a heteroalkyl group, the heteroatom may occupy the position where the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl groups include cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, etc. Examples of heterocyclic alkyl groups include 1-(1,2,5,6-tetrahydropyridinyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, 2-piperazinyl, etc.

[0054] Unless otherwise stated, the terms "halogenated" and "halogen," either on their own or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom. Furthermore, terms such as "halogenated alkyl" are intended to include alkyl groups substituted with 1 to (2m'+1) halogen atoms, which may be the same or different, where m' is the total number of carbon atoms in the alkyl group. For example, the term "halogenated (C1-C4)alkyl" is intended to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, etc. Thus, the term "halogenated alkyl" includes monohalogenated alkyl (an alkyl group substituted with one halogen atom) and polyhalogenated alkyl (an alkyl group substituted with 2 to (2m'+1) halogen atoms, where m' is the total number of carbon atoms in the alkyl group). Unless otherwise stated, the term "per-halogenated alkyl" refers to an alkyl group substituted with (2m'+1) halogen atoms, where m' is the total number of carbon atoms in the alkyl group. For example, the term "fully halogenated (C1-C4) alkyl" is intended to include trifluoromethyl, pentachloroethyl, 1,1,1-trifluoro-2-bromo-2-chloroethyl, etc.

[0055] The term "acyl" refers to the group obtained by removing the hydroxyl group from an organic acid. Therefore, acyl means, for example, acetyl, propionyl, butyryl, decyl, pivaloyl, benzoyl, etc.

[0056] Unless otherwise stated, the term "aryl" means a polyunsaturated hydrocarbon substituent, typically an aromatic hydrocarbon substituent, which can be a monocyclic or a polycyclic (up to three rings) fused together or covalently linked. Non-limiting examples of aryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, and 1,2,3,4-tetrahydronaphthyl.

[0057] The term "heteroaryl" refers to an aryl group (or ring) containing 0-4 heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. The heteroaryl group can be attached to the remainder of the molecule via heteroatoms. Non-limiting examples of heteroaryl groups include 1-pyrrole, 2-pyrrole, 3-pyrrole, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isooxazolyl, 4-isooxazolyl, 5-isooxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furanyl, 3-furanyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, purinel, 2-benzimidazolyl, 5-indolyl, 1-isoquinolinyl, 5-isoquinolinyl, 2-quinoxolinyl, 5-quinoxolinyl, 3-quinolinyl, and 6-quinolinyl.

[0058] For the sake of brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthioxy, aralkyl) includes the aforementioned aryl and heteroaryl rings. Therefore, the term "aralkyl" means those groups comprising an aryl group linked to an alkyl group (e.g., phenyl, phenethyl, pyridylmethyl, etc.), wherein the alkyl group includes those alkyl groups in which a carbon atom (e.g., methylene) is replaced by, for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthoxy)propyl, etc.).

[0059] The terms above (e.g., "alkyl", "heteroalkyl", "aryl", and "heteroaryl") all mean both substituted and unsubstituted forms of the group. Preferred substituents for each type of group are provided below.

[0060] Substituents in alkyl and heteroalkyl groups (as well as those groups referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, ynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be a wide variety of groups, selected from: -OR', =O, =NR', =N-OR', -NR'R", -SR', halogens, -SiR'R"R'", -OC(O)R', -C(O)R', -CO2R', -CONR'R", -OC(O)NR'R", -NR" C(O)R', -NR'-C(O)NR"R'", -NR'-SO2NR'", -NR"CO2R', -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -SO2R', -SO2NR'R", -NR"SO2R, -CN, and -NO2, with 0 to 3 substituents, those having 0, 1, or 2 substituents are particularly preferred. R', R" and R'" each independently refer to hydrogen, unsubstituted (C1-C8) alkyl and heteroalkyl, unsubstituted aryl, aryl substituted with 1 to 3 halogens, unsubstituted alkyl, alkoxy or thioalkoxy, or aryl-(C1-C4) alkyl. When R' and R" are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 5, 6, or 7-membered ring. For example, -NR'R" indicates the inclusion of 1-pyrrolidinyl and 4-morpholinyl. Typically, alkyl or heteroalkyl groups have 0-3 substituents, with those having 2 or fewer substituents being preferred in this invention. More preferably, the alkyl or heteroalkyl group is unsubstituted or monosubstituted. Most preferably, the alkyl or heteroalkyl group is unsubstituted. From the above discussion of substituents, those skilled in the art will understand that the term "alkyl" refers to groups including trihaloalkyl groups (e.g., -CF3 and -CH2CF3).

[0061] Preferred substituents for alkyl and heteroalkyl groups are selected from: -OR', =O, -NR'R", -SR', halogen, -SiR'R"R'", -OC(O)R', -C(O)R', -CO2R', -CONR'R", -OC(O)NR'R", -NR"C(O)R', -NR"CO2R', -NR'-SO2NR"R'", -S(O)R', -SO2R', -SO2NR'R", -NR"SO2R, -CN, and -NO2, wherein R' and R" are defined as above. Further preferred substituents are selected from: -OR', =O, -NR'R", halogen, -OC(O)R', -CO2R', -CONR'R", -OC(O)NR'R", -NR"C(O)R', -NR"CO2R', -NR'-SO2NR"R'", -SO2R', -SO2NR'R", -NR"SO2R, -CN, and -NO2.

[0062] Similarly, the substituents of aryl and heteroaryl groups are also diverse, selected from: halogens, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR"CO2R', -NR'-C(O)NR"R'", -NR'-SO2NR"R'", -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -SO2R', -SO2NR'R", -NR"SO2R, -N3, -CH(Ph)2, perfluoro(C1-C4)alkoxy and perfluoro(C1-C4)alkyl, with the number of substituents ranging from 0 to the open valence of the aromatic ring system. The total number of valences; wherein R', R" and R'" are each independently selected from hydrogen, (C1-C8)alkyl and heteroalkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C1-C4)alkyl and (unsubstituted aryl)oxy-(C1-C4)alkyl. When the aryl group is 1,2,3,4-tetrahydronaphthalene, it may be substituted or unsubstituted with a (C3-C7)spirocycloalkyl group. The (C3-C7)spirocycloalkyl group may be substituted in the same manner as "cycloalkyl" as defined in this invention. Typically, the aryl or heteroaryl group has 0-3 substituents, and those groups having 2 or fewer substituents are preferred in this invention. In one embodiment of the invention, the aryl or heteroaryl group is unsubstituted or monosubstituted. In another embodiment, the aryl or heteroaryl group is unsubstituted.

[0063] Preferred substituents for aryl and heteroaryl groups are selected from: halogens, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -S(O)R', -SO2R', -SO2NR'R", -NR"SO2R, -N3, -CH(Ph)2, perfluorinated (C1-C4) The substituents are alkoxy and perfluorinated (C1-C4)alkyl, wherein R' and R" are defined as above. Further preferred substituents are selected from: halogen, -OR', -OC(O)R', -NR'R", -R', -CN, -NO2, -CO2R', -CONR'R", -NR"C(O)R', -SO2R', -SO2NR'R", -NR"SO2R, perfluorinated (C1-C4)alkoxy and perfluorinated (C1-C4)alkyl.

[0064] The substituent -CO2H used in this invention includes its bioisosteric substitutions; see, for example, The Practice of Medicinal Chemistry; Wermuth, CG, Ed.; Academic Press: New York, 1996; p. 203.

[0065] Two substituents on adjacent atoms of the aromatic or heteroaromatic ring can be optionally replaced by substituents of the formula -TC(O)-(CH2)qU-, where T and U are each independently -NH-, -O-, -CH2-, or a single bond, and q is an integer from 0 to 2. Alternatively, two substituents on adjacent atoms of the aromatic or heteroaromatic ring can be optionally replaced by substituents of the formula -A-(CH2)rB-, where A and B are each independently -CH2-, -O-, -NH-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'-, or a single bond, and r is an integer from 1 to 3. One single bond in the newly formed ring can be optionally replaced by a double bond. Alternatively, the two substituents on adjacent atoms of the aromatic or heteroaromatic ring may be optionally replaced by substituents of the formula -(CH2)sX-(CH2)t-, where s and t are each independently integers from 0 to 3, and X is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. The substituent R' in -NR'- and -S(O)2NR'- is selected from hydrogen or unsubstituted (C1-C6) alkyl groups.

[0066] This invention discloses preferred substituents and provides illustrative examples of preferred substituents in tables, structures, embodiments, and claims. Preferred substituents can be applied cross-referenced to different compounds of this invention; that is, any substituent of a specified compound can be used in combination with other compounds.

[0067] In a particular embodiment, the applicable substituents are each independently a substituted or unsubstituted heteroatom, a substituted or unsubstituted C1-C6 alkyl group containing 0-3 heteroatoms, a substituted or unsubstituted C2-C6 alkenyl group containing 0-3 heteroatoms, a substituted or unsubstituted C2-C6 alkynyl group containing 0-3 heteroatoms, or a substituted or unsubstituted C6-C14 aryl group containing 0-3 heteroatoms, wherein each heteroatom is independently oxygen, phosphorus, sulfur, or nitrogen.

[0068] In more specific embodiments, the applicable substituents are each independently aldehyde, aldehyde imino, alkanoyloxy, alkoxy, alkoxycarbonyl, alkoxy, alkyl, amino, azo, halogen, carbamoyl, carbonyl, formamido, carboxyl, cyano, ester, halogen, haloformyl, hydroperoxyl, hydroxyl, imino, isocyanide, isocyanate, N-tert-butoxycarbonyl, nitrate, nitrile, nitrite, nitro, nitrosyl, phosphate, phosphonoyl, sulfide, sulfonyl, sulfonyl, mercapto, thiol, thiocyanate, trifluoromethyl, or trifluoromethoxy (OCF3).

[0069] The term "pharmaceutically acceptable salt" is intended to include salts of active compounds prepared with relatively non-toxic acids or bases, encompassing specific substituents of compounds according to the invention. When the compounds of the invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compound in its neutral form with a sufficient amount of a pure or suitable inert solvent containing a desired base. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compound in its neutral form with a sufficient amount of a pure or suitable inert solvent containing a desired acid. Pharmaceutically acceptable examples of acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrocarbonic acid, phosphoric acid, monohydrophosphoric acid, dihydrophosphoric acid, sulfuric acid, monohydrosulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, oxalic acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids, such as arginine, and salts of organic acids, such as glucuronic acid or galacturonic acid, are also included. Certain specific compounds of the present invention contain both basic and acidic functional groups, which allows these compounds to be converted into base or acid addition salts.

[0070] The neutral form of the compound can be regenerated by first contacting the salt with a base or acid, and then separating the parent compound using conventional methods. The parent form of the compound differs from the various salt forms in some physical properties, such as solubility in polar solvents; however, in other respects, for the purposes of this invention, these salts are equivalent to the parent form of the compound.

[0071] In addition to salt forms, the present invention also provides compounds in prodrug form. Prodrugs of the compounds described in this invention are those compounds that, after undergoing chemical changes under physiological conditions, provide the compounds of the present invention. Furthermore, in an in vitro environment, prodrugs can be converted into the compounds of the present invention by chemical or biochemical methods. For example, when placed in a reservoir of a transdermal patch with a suitable enzyme or chemical reagent, a prodrug can be slowly converted into the compounds of the present invention. Prodrugs are generally useful because, in some cases, they can be more readily administered than the parent drug. For example, when administered orally, a prodrug can have higher bioavailability than the parent drug. Prodrugs can also have higher solubility in pharmacological compositions than the parent drug. A wide variety of prodrug derivatives are known in the art, such as those dependent on the hydrolytic cleavage or oxidative activation of the prodrug. Non-limiting examples of prodrugs are compounds of the present invention administered as an ester (“prodrug”) followed by hydrolysis and metabolism to the active entity, a carboxylic acid. Other examples include peptide derivatives of the compounds of the present invention.

[0072] Some compounds of the present invention may exist in both solvated and hydrated forms, including hydrated forms. Generally, the hydrated form is equivalent to the hydrated form and is included within the scope of the present invention. Some compounds of the present invention may exist in polycrystalline or amorphous forms. Generally, all physical forms are equivalent for the purposes covered by the present invention and are included within the scope of the present invention.

[0073] Some compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds; racemic, diastereomers, geometric isomers and individual isomers are all included within the scope of the present invention.

[0074] A mixture of diastereomers can be separated into individual diastereomers based on differences in their physicochemical properties using methods well known to those skilled in the art, such as chromatography and / or separation crystallization. Enantiomers can be separated by reacting the enantiomer mixture with a suitable optically active compound (e.g., a chiral aid, such as a chiral alcohol or Mosher's acid chloride) to convert it into a mixture of diastereomers, then separating the diastereomers, and finally converting (e.g., hydrolyzing) each diastereomer to its corresponding pure enantiomer. Enantiomers can also be separated using a chiral HPLC column.

[0075] A single stereoisomer, for example, a substantially pure enantiomer, can be obtained by resolving a racemic mixture using methods such as forming diastereomers with an optically active resolving agent (Eliel, E. and Wilen, S. Stereochemistry of Organic Compounds. New York: John Wiley & Sons, Inc., 1994; Lochmuller, CH, et al. "Chromatographic resolution of enantiomers: Selective review." J. Chromatogr., 113(3)(1975): pp. 283-302). Racemic mixtures of chiral compounds of the present invention can be separated and isolated by any suitable method, including: (1) forming diastereomeric ionic salts with chiral compounds and then separating them by separation crystallization or other methods; (2) forming diastereomeric compounds with chiral derivatizing agents, separating the diastereomers, and converting them into pure stereoisomers; and (3) directly separating substantially pure or enriched stereoisomers under chiral conditions. See: Wainer, Irving W., Ed. Drug Stereochemistry: Analytical Methods and Pharmacology. New York: Marcel Dekker, Inc., 1993.

[0076] The compounds of the present invention may also contain non-natural isotopic atomic moieties on one or more atoms constituting these compounds. For example, the compounds may use radioactive isotopes, such as tritium. 3 H), Iodine-125 ( 125 I) or carbon-14 ( 14 C) Radiolabeling. All isotopic variants of the compounds of this invention, whether or not they are radioactive, are included within the scope of this invention.

[0077] The term "therapeutic effective amount" refers to the amount of a subject compound that, as determined by researchers, veterinarians, physicians, or other clinicians, will elicit a biological or medical response in tissues, the whole body, an animal, or a human to a certain degree. For example, it may be sufficient, upon administration, to prevent or alleviate the development of one or more symptoms of the treated disease or condition. Therapeutic effective amounts will vary depending on the compound, the disease and its severity, and the age, weight, etc., of the mammal being treated.

[0078] The present invention also provides pharmaceutical compositions comprising a subject compound and pharmaceutically acceptable excipients, particularly those compositions comprising a unit dose of the subject compound, especially those compositions packaged together with a specification describing the use of the composition to treat an applicable disease or condition (the present invention).

[0079] Compositions for administration can be in the form of bulk liquid solutions or suspensions, or bulk powders. However, more commonly, compositions are presented in unit dose form for convenient and precise dosing. The term "unit dose form" refers to a physically discrete unit suitable for a single dose for human subjects or other mammals, each unit containing a predetermined amount of active substance calculated to produce the intended therapeutic effect, along with appropriate pharmaceutical excipients. Representative unit dose forms include pre-filled, pre-filled ampoules or syringes of liquid compositions, or pills, tablets, capsules, lozenges, etc., of solid compositions. In these compositions, the compound is often present in trace amounts (from about 0.1% to about 50% by weight, or preferably from about 1% to about 40% by weight), with the remainder being various media or carriers and processing aids that facilitate the formation of the intended dosage form.

[0080] Suitable excipients or carriers, and methods for preparing the dosedable compositions, are well known or obvious to those skilled in the art, and are described in more detail in publications such as Remington's Pharmaceutical Science, Mack Publishing Co, NJ (1991). Furthermore, the compounds can be advantageously used in combination with other therapeutic agents described herein, or those known in the art, particularly other anti-autoimmune agents. Therefore, the compositions can be administered alone, in combination, or in a single dose unit.

[0081] Dosage depends on the formulation of the compound, route of administration, etc., and is usually determined empirically in routine trials, and may vary depending on the target of administration, host, and route of administration. Typically, the amount of active compound in a unit dose formulation can vary or be adjusted from about 1, 3, 10, or 30 to about 30, 100, 300, or 1000 mg depending on the specific application. In a particular embodiment, the unit dose is packaged in a combination pack suitable for sequential use, such as a blister pack containing at least 6, 9, or 12 unit doses. The actual dosage used may vary depending on the patient's needs and the severity of the condition being treated. Determining the correct dosage for a specific situation is within the scope of the art. Typically, the initial dose for treatment is a small dose, less than the optimal dose of the compound. The dose is then gradually increased in small amounts until the optimal effect for that situation is achieved. For convenience, if necessary, the total daily dose may be divided into several portions and administered within one day.

[0082] Compounds can be administered via various methods, including but not limited to parenteral, topical, oral, or local administration, such as via aerosol or transdermal delivery, for prophylactic and / or therapeutic treatment. Furthermore, treatment regimens (e.g., dosage and frequency of administration) can be modified according to the knowledge of a skilled clinician, based on observed effects of the administered therapeutic on the patient and variations in the observed disease response to the administered therapeutic.

[0083] For treating patients, the therapeutic agents of the present invention can be administered at therapeutically effective doses and amounts during an effective treatment regimen. Although the optimal dose is compound-specific for each compound and is generally determined based on practical experience, for more effective compounds, a dose of micrograms (μg) per kilogram of patient body weight is sufficient, for example, in the range of about 1, 10, or 100 μg / kg to about 0.01, 0.1, 1, 10, or 100 mg / kg of patient body weight.

[0084] Generally, routine trials in clinical trials determine the specific range of optimal therapeutic effects for each therapeutic agent and dosing regimen. Dosing to specific patients is also adjusted to an effective and safe range based on the patient's condition and response to the initial dose. However, the final dosing regimen will be adjusted based on the judgment of the participating clinicians based on factors such as the patient's age, condition, and size, as well as the compound potency and the severity of the disease being treated. For example, the compound may be administered orally in 2 to 4 divided doses (preferably 2) of 10 mg to 2000 mg / day, preferably 10 to 1000 mg / day, more preferably 50 to 600 mg / day. Intermittent therapy (e.g., treatment for one week out of every three weeks, or three weeks out of every four weeks) may also be used.

[0085] This invention provides novel solutions for T-cell and B-cell-mediated autoimmune diseases and cancer. Certain autoimmune diseases, such as inflammatory diseases (e.g., inflammatory bowel disease, rheumatoid arthritis, glomerulonephritis and pulmonary fibrosis, psoriasis, allergic skin reactions, atherosclerosis, restenosis, allergic asthma, multiple sclerosis, and type 1 diabetes), are associated with inappropriate T-cell activation (JH Hanke et al., Inflamm. Res., 1995, 357). Furthermore, acute rejection of transplanted organs and graft-versus-host disease (GvHD) after allogeneic bone marrow and stem cell transplantation can also be explained as a result of inappropriate T-cell activation. Dysactivation of B lymphocytes is a hallmark of many autoimmune diseases, such as Hashimoto's thyroiditis, autoimmune hemolytic anemia, autoimmune atrophic gastritis causing pernicious anemia, autoimmune encephalomyelitis, autoimmune orchitis, Goodpasture disease, autoimmune thrombocytopenic purpura including idiopathic thrombocytopenic purpura, sympathetic ophthalmia, myasthenia gravis, Graves' disease, primary biliary cirrhosis, chronic active hepatitis, ulcerative colitis, and membranous nephropathy, as well as those diseases identified as being associated with systemic autoimmune disorders, such as systemic lupus erythematosus, immune thrombocytopenic purpura, rheumatoid arthritis, Sjögren's syndrome, Reiter's syndrome, polymyositis and dermatomyositis, systemic sclerosis, polyarteritis nodosa, multiple sclerosis, and bullous pemphigoid. This invention can also be used to treat other autoimmune diseases based on B cells (humoral) or T cells, including Cogan syndrome, ankylosing spondylitis, Wegener's granulomatosis, autoimmune alopecia, type I or juvenile diabetes, and thyroiditis. Other diseases in which T cell and B cell dysfunction plays an important role are malignancies such as gastrointestinal cancers, colon cancer, liver cancer, skin cancer including mast cell tumors and squamous cell carcinomas, and breast and mammary cancers. Cancer), ovarian cancer, prostate cancer, lymphoma and leukemia (including but not limited to acute myeloid leukemia, chronic myeloid leukemia, mantle cell lymphoma, non-Hodgkin's (NHL) B-cell lymphoma (e.g., precursor B-ALL, marginal zone B-cell lymphoma, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, Burkitt's lymphoma, mediastinal large B-cell lymphoma), Hodgkin's lymphoma, natural killer and T-cell lymphoma; myeloma (including multiple myeloma), myeloproliferative disorders, kidney cancer, lung cancer, muscle cancer, bone cancer, bladder cancer, brain cancer, melanoma (including oral melanoma and metastatic melanoma), Kaposi's sarcoma, proliferative diabetic retinopathy and angiogenesis-related disorders (including solid tumors) and pancreatic cancer [Lim et al, Haematologica, 95 (2010) pp 135-143].We have developed a series of multi-target small molecule kinase inhibitors by focusing on a group of well-established and highly relevant kinase targets associated with the above indications, including but not limited to Lck, Fyn, Btk, and p38 MAPK.

[0086] It should be understood that the embodiments and implementations described in this invention are for illustrative purposes only, and various modifications or changes made thereto are intended to be apparent to those skilled in the art and are included within the spirit and scope of this application and the appended claims. For all purposes, all publications, patents, and patent applications cited in this invention, including their citations, are incorporated herein by reference in their entirety. Example

[0087] reagents and chemicals

[0088] Chromatographic columns used for protein purification: NI-NTA-agarose, MonoQ HP column, and Superdex 75 were purchased from GE Healthcare. Reagents used for crystallization were obtained from Hampton Research. DNA sequences encoding human mitogen-activated protein kinase 14 (hMAPK14) and human mitogen-activated protein kinase 6 (MKK6) were purchased from OpenBiosystems. Unless otherwise specified, all reactions were carried out under inert nitrogen or argon atmosphere. DMF and DMSO (99.9%, extremely dry) were used directly as received. Unless otherwise specified, all other reagents were commercially available and used directly as received. NMR spectra were recorded using a Bruker spectrometer. 1 H (400 MHz) and NMR chemical shifts are reported as values ​​relative to the TMS internal standard (δ = 0.00 ppm) or the residual protonated solvent. Data are presented as follows: chemical shift (ppm), multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, sept = septet, m = multiplet, br = broad peak), coupling constant J (Hz), and integral.

[0089] synthesis:

[0090]

[0091] i) Aminoguanidine nitrate, NaOH, H2O, reflux;

[0092] ii) a) 2-Chloro-6-fluorobenzaldehyde, toluene, reflux; b) NaBH3CN, acetic acid, room temperature.

[0093] Step 1: Synthesis of 3-(furan-2-yl)-1H-1,2,4-triazol-5-amine (1):

[0094] A suspension of aminoguanidine nitrate (22.0 g, 160.5 mmol) in acetonitrile (200 mL) was cooled to 0 °C, and NaOH (20.0 g, 500.0 mmol) was added in portions. The resulting mixture was stirred at room temperature for 3 h. Then, 2-furanoyl chloride (20.0 g, 153.2 mmol) was added dropwise at 0 °C. After the addition was complete, the resulting mixture was stirred at room temperature for 6 h. The solvent was then removed by rotary evaporation, and the residue was dissolved in water (200 mL) and heated under reflux for 5 h. The solvent was removed under reduced pressure, and the crude product was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give 3-(furan-2-yl)-1H-1,2,4-triazol-5-amine (1) (8.5 g, yield 37%) as a white solid. Mass spectrometry (ESI) m / z C6H6N4O[M+H] + Calculated value: 151.05, measured value: 151.10. 1 H NMR (400MHz, d6-DMSO) δ (ppm) 12.07 (s, 1H), 7.68 (s, 1H), 6.71-6.61 (m, 1H), 6.54 (s, 1H), 6.08 (s, 2H).

[0095] Step 2: Synthesis of N-(2-chloro-6-fluorobenzyl)-3-(furan-2-yl)-1H-1,2,4-triazol-5-amine(a):

[0096] A mixture of 3-(furan-2-yl)-1H-1,2,4-triazol-5-amine (1) (8.5 g, 56.6 mmol) and 2-chloro-6-fluorobenzaldehyde (17 g, 107.2 mmol) in anhydrous toluene (100 mL) was heated under reflux for 8 h. After cooling to room temperature, NaBH3CN (5.0 g, 79.6 mmol) and acetic acid (20 mL) were added. The resulting mixture was then stirred at room temperature for 3 h. The reaction mixture was quenched with water (10 mL) and the solvent was removed under reduced pressure. The residue was subjected to reversed-phase rapid column chromatography (C10). 18 Purification yielded N-(2-chloro-6-fluorobenzyl)-3-(furan-2-yl)-1H-1,2,4-triazol-5-amine (a) as a white solid (3.2 g, yield 19.3%). Mass spectrometry (ESI) m / z C 13 H 10 ClFN4O[M+H] + Calculated values: 293.05, 295.05; measured values: 293.2, 295.3. 1H NMR(400MHz,d6-DMSO)δ12.22(s,1H),7.70(s,1H),7.37(dd,J=20.6,7.3Hz,2 H), 7.24 (t, J = 8.7Hz, 1H), 6.94 (s, 1H), 6.73 (s, 1H), 6.55 (s, 1H), 4.50 (s, 2H).

[0097] Step 2: Synthesis of N-(2-chloro-6-methylbenzyl)-3-(furan-2-yl)-1H-1,2,4-triazol-5-amine (b):

[0098] A mixture of 3-(furan-2-yl)-1H-1,2,4-triazol-5-amine (1) (50 mg, 0.33 mmol) and 2-chloro-6-methylbenzaldehyde (97.3 mg, 0.63 mmol) in anhydrous toluene (5 mL) was heated under reflux for 8 h. After cooling to room temperature, NaBH3CN (29.5 mg, 0.47 mmol) and acetic acid (0.5 mL) were added. The resulting mixture was then stirred at room temperature for 3 h. The reaction mixture was quenched with water (1 mL), and the solvent was removed under reduced pressure. The residue was subjected to reversed-phase rapid column chromatography (C1000- ... 18 Purification yielded N-(2-chloro-6-methylbenzyl)-3-(furan-2-yl)-1H-1,2,4-triazol-5-amine (b) as a white solid (12 mg, yield 12.3%). Mass spectrometry (ESI) m / z C 14 H 13 ClFN4O[M+H] + Calculated values: 289.08, 291.07; measured values: 289.40, 291.40. 1 ¹H NMR (400MHz, d6-DMSO / methanol) δ 12.17 (s, 1H), 7.70 (s, 1H), 7.31 (d, J = 7.5Hz, 1H), 7.22 (dd, J = 17.4, 7.1Hz, 2H), 6.82 (s, 1H), 6.73 (s, 1H), 6.56 (s, 1H), 4.48 (d, J = 4.9Hz, 2H), 2.44 (s, 3H).

[0099]

[0100] Step 1: Synthesis of 3-(5-amino-1H-1,2,4-triazol-3-yl)phenol (intermediate 1):

[0101] A suspension of aminoguanidine nitrate (5.46 g, 39.82 mmol) and K₂CO₃ (5.5 g, 39.82 mmol) in N,N-dimethylformamide (DMF) (50 mL) was stirred at room temperature for 1 h. A solution of 3-hydroxybenzoic acid (5.0 g, 36.2 mmol) in N,N-dimethylformamide (DMF) (30 mL) was treated at 0 °C with N,N'-carbonyldiimidazole (CDI) (6.46 g, 39.82 mmol). The resulting solution was then stirred at room temperature for 1 h. This solution was then added dropwise to the suspension. After the addition was complete, the reaction mixture was stirred at room temperature for 3 h. The mixture was then heated to 100 °C and maintained for 5 h. After cooling to room temperature, the mixture was filtered, and the solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give 3-(5-amino-1H-1,2,4-triazol-3-yl)phenol (1) (2.67 g, yield 41.9%) as a white solid. Mass spectrometry (ESI) m / z C8H8N4O[MH] - Calculated value: 175.07, measured value: 175.10. 1 H NMR (400MHz, d6-DMSO) δ 12.06 (s, 1H), 9.47 (s, 1H), 7.36-7.29 (m, 2H), 7.17 (t, J = 8.0Hz, 1H), 6.77-6.67 (m, 1H), 6.00 (s, 2H).

[0102] Step 2: Synthesis of 3-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenol:

[0103] A mixture of 3-(5-amino-1H-1,2,4-triazol-3-yl)phenol (1) (5.0 g, 28.38 mmol), 2-chloro-6-fluorobenzaldehyde (5.0 g, 31.53 mmol), and p-toluenesulfonic acid (500 mg, 2.90 mmol) in isopropanol (50 mL) was heated under reflux overnight. The solution was cooled to room temperature, and then NaBH3CN (4.0 g, 63.65 mmol) and acetic acid (2 mL) were added. The resulting mixture was then stirred at room temperature for 10 h and quenched with water. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give 3-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenol (1.2 g, yield 13.3%) as a white solid, and compound 31 (1.8 g) was recovered. Mass spectrometry (ESI) m / z C 15 H 12 ClFN4O[M+H] +Calculated values: 319.07, 321.07; measured values: 319.40, 321.20. 1 H NMR (400MHz, d6-DMSO) δ12.15(s,1H),9.43(s,1H),7.35(s,4H),7.13-7.27(m,2H),6.90(s,1H),6.75(s,1H),4.51(s,2H).

[0104]

[0105] (i) a) Aminoguanidine nitrate, NaOH, DMF, 0°C; b) CDI, DMF, 0°C to room temperature; c) H2O, reflux;

[0106] (ii)a) 2-Chloro-6-fluorobenzaldehyde, toluene, reflux; b) NaBH3CN, acetic acid, room temperature;

[0107] (iii) BBr3, cyclohexene, DCM, 0℃.

[0108] Step 1: Synthesis of 3-(3,5-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1):

[0109] A suspension of aminoguanidine nitrate (824.7 mg, 6.02 mmol) and NaOH (240.6 mg, 6.02 mmol) in N,N-dimethylformamide (DMF) (10 mL) was cooled to 0 °C. A solution of 3,5-dimethoxybenzoic acid (1.0 g, 5.49 mmol) and N,N-diisopropylethylamine (DIPEA) (780 mg, 6.03 mmol) in N,N-dimethylformamide (DMF) (10 mL) was treated with N,N'-carbonyldiimidazole (CDI) (890 mg, 5.49 mmol) at 0 °C. The resulting solution was then stirred at room temperature for 3 h. This solution was then added dropwise to the suspension. After the addition was complete, the resulting reaction mixture was stirred at room temperature for 3 h. The solvent was then removed by rotary evaporation, and the resulting residue was dissolved in water (70 mL) and heated under reflux for 18 h. After cooling to room temperature, the mixture was filtered, and the resulting solid was recrystallized from methanol to give 3-(3,5-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1) (260 mg, yield 21.5%), a creamy white solid. Mass spectrometry (ESI) m / z C 10 H 12 N4O2[M+H] + Calculated value: 221.10; measured value: 221.40. 1H NMR (400MHz, d6-DMSO) δ 12.08 (s, 1H), 7.05 (dd, J = 10.8, 2.3Hz, 2H), 6.46 (s, 1H), 6.08 (s, 2H), 3.71 (s, 6H).

[0110] Step 2: Synthesis of N-(2-chloro-6-fluorobenzyl)-3-(3,5-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (12):

[0111] A mixture of 3-(3,5-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1) (50 mg, 0.23 mmol) and 2-chloro-6-fluorobenzaldehyde (36 mg, 0.23 mmol) in anhydrous toluene (10 mL) / DMSO (0.1 mL) was heated under reflux for 3 h. The solution was cooled to room temperature, and NaBH3CN (42.8 mg, 0.68 mmol) and acetic acid (1 mL) were added. The resulting mixture was then stirred at room temperature for 10 h and quenched with water. The solvent was removed under reduced pressure. The residue was washed with water (25 mL), and the resulting solid was recrystallized from ethyl acetate / petroleum ether to give N-(2-chloro-6-fluorobenzyl)-3-(3,5-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (12) (30 mg, yield 36.4%) as a white solid. Mass spectrometry (ESI) m / z C 17 H 16 ClFN4O2[M+H] + Calculated values: 363.09, 365.09; measured values: 363.20, 365.20. 1 H NMR(400MHz,d6-DMSO)δ7.97(s,1H),7.51(d,J=8.0Hz,1H),7.41(t,J=8.0Hz,1 H), 7.31 (d, J = 8.2Hz, 1H), 7.12 (s, 2H), 6.57 (s, 1H), 4.67 (s, 2H), 3.80 (s, 6H).

[0112] Step 3: Synthesis of 5-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)benzene-1,3-diol (9):

[0113] A mixture of compound 12 (18 mg, 0.05 mmol) and cyclohexene (2 mL) in dichloromethane (DCM) (10 mL) was cooled to 0 °C. A solution of BBr3 (220 mg, 0.88 mmol) in dichloromethane (DCM) (1 mL) was added dropwise to the mixture. After the addition was complete, the resulting reaction mixture was stirred at 0 °C for 18 h. The solvent was then removed by rotary evaporation, and the residue was analyzed by reversed-phase HPLC (C6000 ppm).18 Purification yielded 5-(5-(((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl-1,3-diol (9) as a white solid (3 mg, yield 18.1%). Mass spectrometry (ESI) m / z C 15 H 12 ClFN4O2[M+H] + Calculated values: 335.06, 337.06; measured values: 335.05, 337.10. 1 H NMR (400MHz, CD3OD) δ7.32 (dt, J = 11.3, 8.1 Hz, 2H), 7.12 (t, J = 8.6 Hz, 1H), 6.85 (s, 2H), 6.32 (s, 1H), 4.64 (s, 2H).

[0114]

[0115] (i) a) Aminoguanidine nitrate, NaOH, methanol, 0°C; b) CDI, DMF, 0°C to room temperature; c) H2O, reflux;

[0116] (ii)a) 2-Chloro-6-fluorobenzaldehyde, toluene, reflux; b) NaBH3CN, acetic acid, room temperature.

[0117] Step 1: Synthesis of 3-(3,4-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1):

[0118] A suspension of aminoguanidine nitrate (13.55 g, 98.83 mmol) and NaOH (240.6 mg, 6.02 mmol) in methanol (100 mL) was stirred at room temperature for 3 h. The solvent was removed by evaporation, and the residue was redissolved in N,N-dimethylformamide (DMF) (100 mL). A solution of 3,4-dimethoxybenzoic acid (15.0 g, 82.34 mmol) in N,N-dimethylformamide (DMF) (150 mL) was treated at 0 °C with N,N'-carbonyldiimidazole (CDI) (14.69 g, 90.59 mmol). The solution was then stirred at room temperature for 3 h. The solution was then added dropwise to the suspension. After the addition was complete, the reaction mixture was stirred at room temperature for 3 h. The solvent was then removed by rotary evaporation; the residue was dissolved in water (150 mL) and heated under reflux for 18 h. After cooling to room temperature, the mixture was filtered, and the resulting solid was recrystallized from methanol to give 3-(3,4-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1) (9.38 g, yield 51.7%), a creamy white solid. Mass spectrometry (ESI) m / z C 10 H 12 N4O2[M+H] +Calculated value: 221.10; measured value: 221.40. 1 HNMR (400MHz, d6-DMSO) δ11.95(s,1H),7.46–7.39(m,2H),6.98(d,J=8.8Hz,1H),5.93(s,2H),3.77(d,J=3.4Hz,6H).

[0119] Step 2: Synthesis of N-(2-chloro-6-fluorophenyl)-3-(3,4-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (11):

[0120] A mixture of 3-(3,4-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1) (8 g, 36.33 mmol), 2-chloro-6-fluorobenzaldehyde (8 g, 50.46 mmol), and p-toluenesulfonic acid (625.5 mg, 3.63 mmol) in isopropanol (40 mL) was heated under reflux overnight. After cooling the solution to room temperature, NaBH3CN (5.0 g, 79.56 mmol) and acetic acid (1 mL) were added. The resulting mixture was then stirred at room temperature for 5 h and quenched with water. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give N-(2-chloro-6-fluorobenzyl)-3-(3,4-dimethoxyphenyl)-1H-1,2,4-triazol-5-amine (11) (4.5 g, yield 34.1%) as a white solid. Mass spectrometry (ESI) m / z C 17 H 16 ClFN4O2[M+H] + Calculated values: 363.09, 365.09; measured values: 363.40, 365.40. 1 ¹H NMR (400MHz, d6-DMSO, 2:1 mixture of rotational isomers) δ 13.13 (s, 0.5H), 12.06 (s, 1H), 7.50–7.41 (m, 3H), 7.40–7.29 (m, 3H), 7.28–7.15 (m, 2H), 7.05 (d, J = 8.2Hz, 0.5H), 6.98 (d, J = 8.8Hz, 1H), 6.87 (t, J = 5.4Hz, 1H), 6.12 (t, J = 6Hz, 0.5H), 4.53 (d, J = 4.9Hz, 2H), 4.45 (d, J = 5.2Hz, 1H), 3.78 (d, J = 6.9Hz, 9H).

[0121]

[0122] Step 1: Synthesis of 3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (intermediate 1):

[0123] A suspension of aminoguanidine nitrate (2.58 g, 18.82 mmol) and NaOH (789.79 mg, 19.75 mmol) in methanol (30 mL) was stirred at room temperature for 3 h. The solvent was evaporated, and the residue was redissolved in N,N-dimethylformamide (DMF) (30 mL). A solution of 3-nitrobenzoic acid (3.0 g, 17.95 mmol) and N,N-diisopropylethylamine (DIPEA) (2.55 g, 19.73 mmol) in N,N-dimethylformamide (DMF) (30 mL) was treated at 0 °C with N,N'-carbonyldiimidazole (CDI) (3.2 g, 19.73 mmol). The resulting solution was stirred at room temperature for 3 h. The solution was then added dropwise to the suspension. After the addition was complete, the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by rotary evaporation, and the residue was dissolved in water (100 mL) and heated under reflux for 18 h. After cooling to room temperature, the mixture was filtered, and the resulting solid was recrystallized from methanol to give 3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (1) (3.0 g, yield 81.5%) as a yellow solid. Mass spectrometry (ESI) m / z C8H7N5O2[M+H] + Calculated value: 206.06, measured value: 206.20. 1 H NMR (400MHz, d6-DMSO) δ12.38 (s, 1H), 8.65–8.60 (m, 1H), 8.28 (d, J = 7.8Hz, 1H), 8.22–8.16 (m, 1H), 7.71 (t, J = 8.0Hz, 1H), 6.22 (s, 2H).

[0124] Step 2: Synthesis of N-(2-chloro-6-fluorobenzyl)-3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (13):

[0125] A mixture of 3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (1) (300 mg, 1.46 mmol) and 2-chloro-6-fluorobenzaldehyde (300 mg, 1.89 mmol) in anhydrous toluene (10 mL) / DMSO (0.1 mL) was heated under reflux for 15 h. After cooling the solution to room temperature, NaBH3CN (200 mg, 3.18 mmol) and acetic acid (1 mL) were added. The resulting mixture was then stirred at room temperature for 3 h and quenched with water. The solvent was removed under reduced pressure, and the residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give N-(2-chloro-6-fluorobenzyl)-3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (2a) (150 mg, yield 29.5%) as a pale yellow solid. Mass spectrometry (ESI) m / z C15 H 11 ClFN5O2[M+H] + Calculated values: 348.06, 350.06; measured values: 348.10, 350.10. 1 H NMR(400MHz,CD3OD)δ8.82(s,1H),8.34(d,J=7.8Hz,1H),8.29–8.22(m,1H), 7.69(t,J=8.0Hz,1H),7.40–7.27(m,2H),7.14(t,J=8.8Hz,1H),4.70(s,2H).

[0126] Step 3: Synthesis of 3-(3-aminophenyl)-N-(2-chloro-6-fluorobenzyl)-1H-1,2,4-triazol-5-amine (16):

[0127] A mixture of N-(2-chloro-6-fluorophenyl)-3-(3-nitrobenzyl)-1H-1,2,4-triazol-5-amine (2a) (50 mg, 0.14 mmol) and hydrazine hydrate (0.5 mL, 80% aqueous solution) in ethanol (10 mL) was heated to 60 °C. Raney-Ni (10 mg) was then added. The resulting mixture was stirred at this temperature for 3 h. After cooling to room temperature, the mixture was filtered, and the solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give 3-(3-aminophenyl)-N-(2-chloro-6-fluorobenzyl)-1H-1,2,4-triazol-5-amine (3a) (36 mg, yield 78.8%) as a white solid. Mass spectrometry (ESI) m / z C 15 H 13 ClFN5[M+H] + Calculated values: 318.08, 320.08; measured values: 318.10, 320.10. 1 H NMR (400MHz, CD3OD) δ7.33(ddd,J=19.0,9.5,6.9Hz,2H),7.27–7.19(m,2H),7.19–7.08(m,2H),6.78(d,J=7.8Hz,1H),4.64(s,2H).

[0128] Synthesis of N-(2-chloro-6-methylbenzyl)-3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (15)

[0129] A mixture of 3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (1) (100 mg, 0.49 mmol) and 2-chloro-6-methylbenzaldehyde (150.69 mg, 0.97 mmol) in anhydrous toluene (6 mL) / ethanol (0.5 mL) was heated under reflux for 15 h. After cooling to room temperature, NaBH3CN (100 mg, 1.59 mmol) and acetic acid (1 mL) were added at 0 °C. The resulting mixture was then stirred at room temperature for 3 h. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give N-(2-chloro-6-methylbenzyl)-3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (2b) (34 mg, yield 20.3%) as a pale yellow solid, and 3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (1) (40 mg) was recovered. Mass spectrometry (ESI) m / z C 16 H 14 ClN5O2[M+H] + Calculated values: 344.08, 346.08; measured values: 344.40, 346.40. 1 H NMR(400MHz,CD3OD)δ8.84–8.81(m,1H),8.37–8.33(m,1H),8.29–8.24(m,1H),7 .70(t,J=8.0Hz,1H),7.30(m,1H),7.22–7.19(m,2H),4.68(s,2H),2.52(s,3H).

[0130] Step 3: Synthesis of 3-(3-aminophenyl)-N-(2-chloro-6-methylphenyl)-1H-1,2,4-triazol-5-amine (19)

[0131] A mixture of compound N-(2-chloro-6-methylbenzyl)-3-(3-nitrophenyl)-1H-1,2,4-triazol-5-amine (2b) (30 mg, 0.087 mmol) and hydrazine hydrate (0.3 mL, 80% aqueous solution) in ethanol (5 mL) was heated to 60 °C. Raney-Ni (10 mg) was then added. The resulting mixture was stirred at this temperature for 3 h. After cooling to room temperature, the mixture was filtered, and the solvent was removed under reduced pressure. The residue was subjected to reversed-phase rapid column chromatography (C10). 18 Purification yielded 3-(3-aminophenyl)-N-(2-chloro-6-methylphenyl)-1H-1,2,4-triazol-5-amine (3b) as a white solid (20 mg, 73% yield). Mass spectrometry (ESI) m / z C 16 H 16 ClN5[M+H] +Calculated values: 314.11, 316.11; measured values: 314.40, 316.40. 1 H NMR (400MHz, CD3OD) δ7.28(m,3H),7.19(m,3H),6.79(s,1H),4.62(s,2H),2.50(s,3H).

[0132] Step 4: Synthesis of N-(3-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)acrylamide (17):

[0133] A mixture of compound 3a (12.0 mg, 0.038 mmol) and triethylamine (5.0 mg, 0.049 mmol) in tetrahydrofuran (THF) (0.5 mL) was cooled to 0 °C, and acryloyl chloride (3.45 mg, 0.038 mmol) was added. The resulting mixture was then stirred at room temperature for 3 h, followed by quenching with water. The solvent was removed under reduced pressure, and the residue was analyzed by reversed-phase HPLC (C6000 ppm). 18 Purification yielded N-(3-(5-(((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)acrylamide (a) as a white solid (2.6 mg, yield 18.5%). Mass spectrometry (ESI) m / z C 18 H 15 ClFN5O[M+H] + Calculated values: 372.09, 374.09; measured values: 372.30, 374.20. 1 H NMR (400MHz, CD3OD) δ8.12(s,1H),7.77(s,1H),7.73–7.63(m,1H),7.40(s,1H),7.37–7.27(m,2H),7.14(t,J=8 .1Hz, 1H), 6.46 (dd, J=17.0, 9.6Hz, 1H), 6.38 (dd, J=17.0, 2.3Hz, 1H), 5.79 (dd, J=9.6, 2.3Hz, 1H), 4.67 (s, 2H).

[0134] Step 4: Synthesis of N-(3-(5-((2-chloro-6-methylbenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)acrylamide (20):

[0135] A mixture of compound 3b (4.0 mg, 0.013 mmol) and triethylamine (30.0 mg, 0.3 mmol) in tetrahydrofuran (THF) (0.5 mL) was cooled to 0 °C, and acryloyl chloride (2.0 mg, 0.022 mmol) was added. The resulting mixture was then stirred at room temperature for 3 h, followed by quenching with water. The solvent was removed under reduced pressure. The residue was analyzed by reversed-phase HPLC (C6000 ppm). 18 Purification yielded N-(3-(5-((2-chloro-6-methylbenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)acrylamide (b(3.1 mg, yield 66.1%)) as a white solid. Mass spectrometry (ESI) m / z C 19 H 18 ClN5O[M+H] + Calculated values: 368.12, 370.12; measured values: 368.40, 370.40. 1 H NMR(400MHz,d6-DMSO)δ10.25(s,1H),8.19(s,1H),7.75(d,J=9.0Hz,1H),7 .62(d,J=7.9Hz,1H),7.36(d,J=7.9Hz,1H),7.32(dd,J=6.8,4.8Hz,1H),7.2 2(dd,J=12.1,4.7Hz,2H),6.46(dd,J=17.0,10.1Hz,1H),6.27(dd,J=17.0,2 .0Hz, 1H), 5.76 (dd, J=10.1, 2.0Hz, 1H), 4.52 (d, J=5.4Hz, 2H), 2.48 (s, 3H).

[0136] Step 4: Synthesis of N-(3-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)-2-cyano-4,4-dimethylpent-2-enamide (42):

[0137] A suspension of pivalaldehyde (202.5 mg, 1.79 mmol, 75% in n-butanol), 2-cyanoacetic acid (150.0 mg, 1.74 mmol), and KOH (200.0 mg, 3.56 mmol) in methanol (2 mL) was heated to 40 °C for 5 h. After TLC analysis showed that the starting material had been consumed, the pH of the mixture was adjusted to 3–4 with 3 M HCl, and extracted with ethyl acetate (50 mL). The resulting ethyl acetate solution was dried and concentrated to give 2-cyano-4,4-dimethylpentan-2-enoic acid (100 mg, yield 37.3%) as a white solid. Mass spectrometry (ESI) m / z C8H 11 NO2[MH] -Calculated value: 152.08, measured value: 152.20. 1 H NMR (400MHz, CDCl3): (Z)-2-cyano-4,4-dimethylpent-2-enoic acid δ4.35(s, 1H), 0.90(s, 9H); (E)-2-cyano-4,4-dimethylpent-2-enoic acid δ7.73(s, 1H), 1.33(s, 9H).

[0138] A mixture of compound 3a (30.0 mg, 0.094 mmol), 2-cyano-4,4-dimethylpentan-2-enoic acid (15.0 mg, 0.098 mmol), and N,N-dimethylpyridin-4-amine (DMAP) (2 mg, 0.016 mmol) in N,N-dimethylformamide (DMF) (2 mL) was cooled to 0 °C, and EDCI (40.0 mg, 0.209 mmol) was added. The resulting mixture was then stirred at room temperature for 20 h and quenched with water. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give N-(3-(5-(((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)-2-cyano-4,4-dimethylpentan-2-enamide (c) (14 mg, yield 32.7%) as a white solid. Mass spectrometry (ESI) m / z C 23 H 22 ClFN6O[M+H] + Calculated values: 453.15, 455.15; measured values: 453.20, 455.30.

[0139] Step 4: Synthesis of N-(3-(5-((2-chloro-6-methylbenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)-2-cyano-4,4-dimethylpent-2-enamide (23):

[0140] A mixture of compound 3b (10.0 mg, 0.032 mmol), 2-cyano-4,4-dimethylpentan-2-enoic acid (5.0 mg, 0.033 mmol), and N,N-dimethylpyridin-4-amine (DMAP) (0.4 mg, 0.0032 mmol) in N,N-dimethylformamide (DMF) (2 mL) was cooled to 0 °C, and EDCI (15.0 mg, 0.078 mmol) was added. The resulting mixture was then stirred at room temperature for 20 h and quenched with water. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give N-(3-(5-(((2-chloro-6-methylbenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)-2-cyano-4,4-dimethylpentan-2-enamide (d) (5.0 mg, 35% yield) as a white solid. Mass spectrometry (ESI) m / z C 24 H 25 ClN6O[M+H] + Calculated values: 449.18, 451.18; measured values: 449.50, 451.30. 1 ¹H NMR (400MHz, CDCl₃, mixture of rotational isomers) δ 7.94 (s, 1H), 7.84 (d, J = 7.7Hz, 1H), 7.79 (s, 0.2H), 7.42 (t, J = 7.8Hz, 1H), 7.32 (d, J = 8.7Hz, 1H), 7.23 (d, J = 7.3Hz, 1H), 7.15–7.09 (m, 2H), 5.05 (s, 1H), 4.67 (s, 2H), 4.35 (d, J = 2.8Hz, 1H), 3.81 (d, J = 2.9Hz, 1H), 2.51 (s, 3H), 1.33 (s, 1.3H), 1.03 (s, 9H).

[0141] Step 4: Synthesis of N-(3-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)but-2-yneamide (37):

[0142] A mixture of compound 3a (15.0 mg, 0.047 mmol), 2-butynedic acid (10.0 mg, 0.118 mmol), and N,N-dimethylpyridin-4-amine (DMAP) (2 mg, 0.016 mmol) in N,N-dimethylformamide (DMF) (2 mL) was cooled to 0 °C, and EDCI (40.0 mg, 0.209 mmol) was added. The resulting mixture was then stirred at room temperature for 20 h and quenched with water. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give N-(3-(5-(((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)but-2-yneamide (5 mg, yield 27.6%) as a white solid. Mass spectrometry (ESI) m / z C 19 H 15 ClFN5O[M+H] + Calculated values: 384.09, 386.09; measured values: 384.30, 86.43. 1 H NMR (400MHz, CD3OD) δ8.05(s,1H),7.68-7.66(m,2H),7.38-7.29(m,3H),7.20-7.13(m,1H),4.67(s,2H),2.04(s,3H).

[0143]

[0144] Step 1: Synthesis of 3-((tert-butoxycarbonyl)amino)benzoic acid (intermediate 1):

[0145] A mixture of 3-aminobenzoic acid (500 mg, 3.65 mmol) and triethylamine (1.11 g, 10.97 mmol) in methanol (6 mL) was cooled to 0 °C, and di-tert-butyl dicarbonate (880 mg, 4.03 mmol) was added. The resulting mixture was then stirred at room temperature for 10 h, followed by quenching with water. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give 3-((tert-butoxycarbonyl)amino)benzoic acid (1) (800 mg, yield 92.5%) as a white solid. Mass spectrometry (ESI) m / z C 12 H 15 NO4[MH] - Calculated value: 236.10, measured value: 236.10. 1H NMR (400MHz, d6-DMSO) δ12.90(s,1H),9.55(s,1H),8.14(s,1H),7.62(d,J=8.5Hz,1H),7.53(d,J=7.8Hz,1H),7.36(t,J=7.9Hz,1H),1.48(s,9H).

[0146] Step 2: Synthesis of 3-((tert-butoxycarbonyl)(methyl)amino)benzoic acid (intermediate 2):

[0147] A mixture of 3-((tert-butoxycarbonyl)amino)benzoic acid (1) (1.0 g, 4.21 mmol) in N,N-dimethylformamide (DMF) (10 mL) was cooled to 0 °C, and sodium hydride (505.7 mg, 12.64 mmol, 60%, dispersed in mineral oil) was added. The mixture was stirred at this temperature for 1 h, and iodomethane (1.26 g, 8.80 mmol) was added. The resulting mixture was stirred at room temperature for 5 h, and then quenched with water. The solvent was removed under reduced pressure, and the residue was dissolved in methanol (10 mL) / water (1 mL), and sodium hydroxide (337.2 mg, 8.43 mmol) was added. After TLC analysis showed that the starting material had been consumed, the pH of the mixture was adjusted to 3–4 with 3 M HCl, and then extracted with ethyl acetate (50 mL). The resulting ethyl acetate solution was dried and concentrated to give 3-((tert-butoxycarbonyl)(methyl)amino)benzoic acid (2) (950 mg, yield 89.7%) as a white solid. Mass spectrometry (ESI) m / z C 13 H 17 NO4[MH] - Calculated value: 250.12; measured value: 250.10. 1 H NMR(400MHz,d6-DMSO)δ13.06(s,1H),7.83(t,J=1.8Hz,1H),7.75–7.72(m,1H), 7.53(ddd,J=8.0,2.3,1.2Hz,1H),7.49–7.43(m,1H),3.20(s,3H),1.40(s,9H).

[0148] Step 3: Synthesis of (3-(5-amino-1H-1,2,4-triazol-3-yl)phenyl)(methyl)carbamate tert-butyl ester (intermediate 3):

[0149] A suspension of aminoguanidine nitrate (360 mg, 2.63 mmol) and K₂CO₃ (660 mg, 4.78 mmol) in N,N-dimethylformamide (DMF) (3 mL) was stirred at room temperature for 1 h. A solution of 3-((tert-butoxycarbonyl)(methyl)amino)benzoic acid (2) (600 mg, 2.39 mmol) in N,N-dimethylformamide (DMF) (30 mL) was treated at 0 °C with N,N'-carbonyldiimidazole (CDI) (426 mg, 2.63 mmol). The solution was stirred at room temperature for 1 h. It was then added to the suspension. The resulting mixture was stirred at room temperature for 3 h, then heated to 100 °C and maintained for 3 h. The mixture was cooled to room temperature, filtered, and concentrated. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give tert-butyl (3-(5-amino-1H-1,2,4-triazol-3-yl)phenyl)(methyl)carbamate (3) as a yellow oil (220 g, yield 31.8%). Mass spectrometry (ESI) m / z C 14 H 19 N5O2[M+H] + Calculated value: 290.15; Measured value: 290.20. 1 H NMR (400MHz, CDCl3) δ7.85–7.80(m,1H),7.73–7.67(m,1H),7.61(s,1H),7.2 9(t,J=7.9Hz,1H),7.23–7.18(m,1H),7.04(s,2H),3.23(s,3H),1.46(s,9H).

[0150] Step 4: Synthesis of tert-butyl (3-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)(methyl)carbamate (intermediate 4):

[0151] A mixture of tert-butyl (3-(5-amino-1H-1,2,4-triazol-3-yl)phenyl)(methyl)carbamate (3) (200 mg, 0.69 mmol), 2-chloro-6-fluorobenzaldehyde (200 mg, 1.26 mmol), and p-toluenesulfonic acid (11.9 mg, 0.069 mmol) in isopropanol (5 mL) was heated under reflux overnight. After cooling the solution to room temperature, NaBH3CN (120 mg, 1.91 mmol) and acetic acid (0.5 mL) were added. The resulting mixture was then stirred at room temperature for 5 h and quenched with water. The solvent was removed under reduced pressure. The crude product of tert-butyl (3-(5-((2-chloro-6-fluorophenyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)(methyl)carbamate (4) was used directly without further purification.

[0152] Step 5: Synthesis of N-(2-chloro-6-fluorobenzyl)-3-(3-(methylamino)phenyl)-1H-1,2,4-triazol-5-amine (18):

[0153] A mixture of crude (3-(5-(((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)(methyl)carbamate tert-butyl ester (4) (100 mg) in tetrahydrofuran (THF) (5 mL) was cooled to 0 °C, and trifluoroacetic acid (10 mL) was added. The resulting mixture was then stirred at room temperature for 5 h. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give N-(2-chloro-6-fluorobenzyl)-3-(3-(methylamino)phenyl)-1H-1,2,4-triazol-5-amine (5) (45 mg, yield 19.6%, two steps) as a white solid. Mass spectrometry (ESI) m / z C 16 H 15 ClFN5[M+H] + Calculated values: 332.10, 334.10; measured values: 332.40, 334.30. 1 ¹H NMR (400MHz, d6-DMSO, 2:1 mixture of rotational isomers) δ 13.17 (s, 0.5H), 12.07 (s, 1H), 7.35 (s, 3H), 7.23 (s, 1.5H), 7.12 (s, 4.5H), 6.85 (s, 1H), 6.48–6.52 (m, 1.5H), 6.13 (s, 0.6H), 5.71–5.75 (m, 1.5H), 4.51 (s, 3H), 2.69 (s, 4.5H).

[0154] Step 6: Synthesis of N-(3-(5-((2-chloro-6-fluorophenyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)-N-methylacrylamide (21)

[0155] A mixture of compound 29 (32 mg, 0.096 mmol) and N-ethyl-N-isopropyl-2-propylamine (DIPEA) (50 mg, 0.39 mmol) in tetrahydrofuran (THF) (0.5 mL) was cooled to 0 °C, and acryloyl chloride (20 mg, 0.22 mmol) was added. The resulting mixture was then stirred at room temperature for 3 h. The solvent was removed under reduced pressure. The residue was dissolved in methanol (2 mL) / water (0.5 mL), and NaOH (10 mg, 0.25 mmol) was added. The reaction mixture was stirred at room temperature for 5 h and concentrated. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give N-(3-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)phenyl)-N-methylacrylamide (6.3 mg, yield 17%) as a white solid. Mass spectrometry (ESI) m / z C 19 H 17 ClFN5O[M+H] + Calculated values: 386.11, 388.11; measured values: 386.50, 388.40. 1 H NMR (400MHz, CDCl3) δ7.95(d,J=7.9Hz,1H),7.83(t,J=1.7Hz,1H),7.43(t,J=7.8Hz,1H),7.25–7.19(m,2H),7.18–7.14(m,1H),7.00-7.05(m,1 H), 6.34 (dd, J=16.8, 1.9Hz, 1H), 6.09 (dd, J=16.5, 10.3Hz, 1H), 5.50 (dd, J=10.3, 1.6Hz, 1H), 5.37 (s, 1H), 4.68 (d, J=5.3Hz, 2H), 3.36 (s, 3H).

[0156]

[0157] Step 1: Synthesis of 3-(4-methoxy-3-nitrophenyl)-1H-1,2,4-triazol-5-amine (intermediate 1):

[0158] A suspension of aminoguanidine nitrate (139 mg, 1.01 mmol) and KOH (57 mg, 1.02 mmol) in methanol (2 mL) was concentrated by stirring at room temperature for 1.5 h. The residue was diluted with N,N-dimethylformamide (DMF) (2 mL). A solution of 4-methoxy-3-nitrobenzoic acid (197 mg, 1.0 mmol) in N,N-dimethylformamide (DMF) (5 mL) was treated with N,N'-carbonyldiimidazole (CDI) (190 mg, 1.17 mmol) at 0 °C. The solution was stirred at room temperature for 1.5 h. Then it was added dropwise to the suspension. After the addition was complete, the reaction mixture was stirred at room temperature for 2 h. The solvent was removed by rotary evaporation, and the residue was dissolved in water (50 mL) and heated under reflux for 5 h. After cooling to room temperature, the mixture was filtered, and the resulting solid was recrystallized from methanol to give 3-(4-methoxy-3-nitrophenyl)-1H-1,2,4-triazol-5-amine (1) as a pale yellow solid (110 mg, yield 46.8%). Mass spectrometry (ESI) m / z C9H9N5O3[M+H] + Calculated value: 236.07; Measured value: 236.20. 1 H NMR (400MHz, d6-DMSO) δ12.15 (s, 1H), 8.26 (d, J = 1.4Hz, 1H), 8.11 (dd, J = 8.8, 2.1Hz, 1H), 7.42 (d, J = 8.9Hz, 1H), 6.14 (s, 2H), 3.95 (s, 3H).

[0159] Step 2: Synthesis of N-(2-chloro-6-fluorobenzyl)-3-(4-methoxy-3-nitrophenyl)-1H-1,2,4-triazol-5-amine (intermediate 2):

[0160] A mixture of 3-(4-methoxy-3-nitrophenyl)-1H-1,2,4-triazol-5-amine (1) (100 mg, 0.43 mmol) and 2-chloro-6-fluorobenzaldehyde (150 mg, 0.95 mmol) in anhydrous toluene (10 mL) was heated under reflux for 15 h. After cooling the solution to room temperature, NaBH3CN (45 mg, 0.72 mmol) and acetic acid (1 mL) were added. The resulting mixture was then stirred at room temperature for 3 h and quenched with water. The solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give N-(2-chloro-6-fluorophenyl)-3-(4-methoxy-3-nitrophenyl)-1H-1,2,4-triazol-5-amine (2) (70 mg, yield 43.6%) as a pale yellow solid, which was used directly in the next step. Mass spectrometry (ESI) m / z C 16 H 13 ClFN5O3[M+H] +Calculated values: 378.07, 380.07; measured values: 378.30, 380.30.

[0161] Step 3: Synthesis of 3-(3-amino-4-methoxyphenyl)-N-(2-chloro-6-fluorobenzyl)-1H-1,2,4-triazol-5-amine (24):

[0162] A mixture of N-(2-chloro-6-fluorobenzyl)-3-(4-methoxy-3-nitrophenyl)-1H-1,2,4-triazol-5-amine (2) (70 mg, 0.19 mmol) and hydrazine hydrate (1.0 mL, 80% aqueous solution) in ethanol (10 mL) was heated to 60 °C. Raney-Ni (10 mg) was then added. The resulting mixture was stirred at room temperature for 3 h. After cooling to room temperature, the mixture was filtered, and the solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel, dichloromethane / methanol = 50:1 to 10:1) to give 3-(3-amino-4-methoxyphenyl)-N-(2-chloro-6-fluorophenyl)-1H-1,2,4-triazol-5-amine (3) (50 mg, yield 77.6%) as a white solid. Mass spectrometry (ESI) m / z C 16 H 15 ClFN5O[M+H] + Calculated values: 348.09, 350.09; measured values: 348.50, 350.60. 1 H NMR (400MHz, CD3OD) δ7.36–7.23(m,4H),7.11(t,J=8.3Hz,1H),6.89(d,J=8.2Hz,1H),4.63(s,2H),3.88(s,3H).

[0163] Step 4: Synthesis of N-(5-(5-((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)-2-methoxyphenyl)acrylamide (22):

[0164] A mixture of compound 3 (8.0 mg, 0.023 mmol) and N-ethyl-N-isopropyl-2-propylamine (DIPEA) (12.0 mg, 0.093 mmol) in tetrahydrofuran (THF) (0.5 mL) was cooled to 0 °C, and acryloyl chloride (5.0 mg, 0.055 mmol) was added. The resulting mixture was stirred at room temperature for 30 min, followed by quenching with water. The solvent was removed under reduced pressure. The residue was dissolved in methanol (2 mL) / water (0.5 mL), and then NaOH (5.0 mg, 0.13 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The residue was concentrated and purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give N-(5-(5-(((2-chloro-6-fluorobenzyl)amino)-1H-1,2,4-triazol-3-yl)-2-methoxyphenyl)-acrylamide (5.1 mg, yield 55.2%) as a white solid. Mass spectrometry (ESI) m / z C 19 H 17 ClFN5O2[MH] - Calculated values: 400.11, 401.11; measured values: 400.40, 402.40. 1 H NMR (400MHz, CDCl3) δ8.94(s,1H),7.91(s,1H),7.73(dd,J=8.5,2.1Hz,1H),7.21-7.16(m,2H),7.03-6.97(m,1H),6.94(d,J=8.6Hz,1H),6 .42(dd,J=16.9,1.4Hz,1H),6.30(dd,J=16.9,10.1Hz,1H),5.77(dd,J=10.0,1.4Hz,1H),5.09(s,1H),4.68(d,J=5.0Hz,2H),3.94(s,3H).

[0165]

[0166] Step 1: Synthesis of 3-(3,4,5-trimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1):

[0167] A suspension of aminoguanidine nitrate (137 mg, 1.0 mmol) and K₂CO₃ (276 mg, 2.0 mmol) in N,N-dimethylformamide (DMF) (3 mL) was stirred at room temperature for 1 h. A solution of 3,4,5-trimethoxybenzoic acid (212 mg, 1.0 mmol) in N,N-dimethylformamide (DMF) (5 mL) was treated with N,N'-carbonyldiimidazole (CDI) (162 mg, 1.0 mmol) at 0 °C. The solution was stirred at room temperature for 1 h. It was then added to the suspension. The resulting mixture was stirred at room temperature for 3 h, then heated to 100 °C and maintained for 3 h. The mixture was cooled to room temperature, filtered, and concentrated to give crude 3-(3,4,5-trimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1) (180 mg) as a white solid, which was used directly in the next step. Mass spectrometry (ESI) m / z C 11 H 14 N4O3[M+H] + Calculated value: 251.11; Measured value: 241.20.

[0168] Step 2: Synthesis of N-(2-chloro-6-fluorobenzyl)-3-(3,4,5-trimethoxyphenyl)-1H-1,2,4-triazol-5-amine (36):

[0169] A mixture of 3-(3,4,5-trimethoxyphenyl)-1H-1,2,4-triazol-5-amine (intermediate 1) (180 mg, 0.72 mmol), 2-chloro-6-fluorobenzaldehyde (180 mg, 1.14 mmol), and p-toluenesulfonic acid (20.00 mg, 0.12 mmol) in isopropanol (5 mL) was heated under reflux overnight. After cooling the solution to room temperature, NaBH3CN (180 mg, 2.86 mmol) and acetic acid (1.5 mL) were added. The resulting mixture was then stirred at room temperature for 5 h and quenched with water. The solvent was removed under reduced pressure, and the residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give N-(2-chloro-6-fluorophenyl)-3-(3,4,5-trimethoxyphenyl)-1H-1,2,4-triazol-5-amine (65 mg, yield 16.6%, 2 steps) as a white solid. Mass spectrometry (ESI) m / z C 18 H 18 ClFN4O3[M+H] + Calculated values: 393.11, 395.11; measured values: 393.33, 395.27. 1H NMR(400MHz, CDCl3)δ7.96(s,1H),7.25-7.22(m,1H),7.18(s,2H),7.15-7.11(m,1H),7. 06-7.02(m,1H),5.22(t,J=5.6Hz,1H),4.65(d,J=5.6Hz,2H),3.89(s,6H),3.87(s,3H).

[0170]

[0171] Step 1: Synthesis of 3-(1H-benzo[d]imidazol-2-yl)-1H-1,2,4-triazol-5-amine (intermediate 1):

[0172] A suspension of aminoguanidine nitrate (150 mg, 1.09 mmol) and K₂CO₃ (150 mg, 1.09 mmol) in N,N-dimethylformamide (DMF) (3 mL) was stirred at room temperature for 1 h. A solution of 1H-benzo[d]imidazole-2-carboxylic acid (162 mg, 1.0 mmol) in N,N-dimethylformamide (DMF) (5 mL) was treated at 0 °C with N,N'-carbonyldiimidazole (CDI) (180 mg, 1.1 mmol). The solution was stirred at room temperature for 1 h. It was then added to the suspension. The resulting mixture was stirred at room temperature for 3 h, then heated to 100 °C and maintained for 3 h. The mixture was cooled to room temperature, filtered, and concentrated to give crude 3-(1H-benzo[d]imidazole-2-yl)-1H-1,2,4-triazol-5-amine (intermediate 1) (290 mg, purity approximately 40%) as a white solid, which was used directly in the next step. Mass spectrometry (ESI) m / z C9H8N6 [M+H] + Calculated value: 201.08, measured value: 201.20.

[0173] Step 2: Synthesis of 3-(1H-benzo[d]imidazol-2-yl)-N-(2-chloro-6-fluorobenzyl)-1H-1,2,4-triazol-5-amine (31):

[0174] A mixture of 3-(1H-benzo[d]imidazol-2-yl)-1H-1,2,4-triazol-5-amine (intermediate 1) (50 mg, 0.25 mmol, 40% purity), 2-chloro-6-fluorobenzaldehyde (50 mg, 0.32 mmol), and p-toluenesulfonic acid (5.00 mg, 0.03 mmol) in ethanol (5 mL) was heated under reflux overnight. After cooling the solution to room temperature, NaBH3CN (50 mg, 0.80 mmol) and acetic acid (1.5 mL) were added. The resulting mixture was then stirred at room temperature for 5 h and quenched with water. The solvent was removed under reduced pressure, and the residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give 3-(1H-benzo[d]imidazol-2-yl)-N-(2-chloro-6-fluorobenzyl)-1H-1,2,4-triazol-5-amine (1.4 mg) as a white solid. Mass spectrometry (ESI) m / z C 16 H 12 ClFN6[M+H] + Calculated values: 343.08, 345.08; measured values: 343.33, 345.27. 1 H NMR (400MHz, CD3OD) δ7.62(s,2H),7.32-7.27(m,4H),7.20-7.14(m,1H),4.72(s,2H).

[0175]

[0176] Step 1: Synthesis of 3,4-bis(2-methoxyethoxy)benzoic acid (intermediate 1):

[0177] A suspension of 3,4-dihydroxybenzoic acid (1.0 g, 6.49 mmol), TBAI (239.67 mg, 0.65 mmol), and K₂CO₃ (3.59 g, 25.98 mmol) in N,N-dimethylformamide (DMF) (10 mL) was heated to 100 °C and maintained for 1 h. After cooling the mixture to 50 °C, 1-chloro-2-methoxyethane (2.45 g, 25.92 mmol) was added. The resulting mixture was stirred at 85 °C for 15 h, then cooled to room temperature. The mixture was filtered, concentrated, and the residue was dissolved in THF (40 mL) / H₂O (10 mL) with KOH (1.09 g, 19.5 mmol). The resulting solution was stirred at room temperature for 5 h, then quenched with 1N HCl. Extracted again with ethyl acetate (30 mL, 3 times), the organic phase was washed with brine (15 mL), dried on Na2SO4, filtered, concentrated, and 3,4-bis(2-methoxyethoxy)benzoic acid (intermediate 1) (1.32 g, yield 75.33%) was obtained as a clear oil. 1H NMR (400MHz, CDCl3) δ7.48 (d, J=8.4Hz, 1H), 7.65 (s, 1H), 6.94 (d, J=8.4Hz, 1H), 4.24-4.16 (m, 4H), 3.82-3.76 (m, 4H), 3.47 (s, 6H).

[0178] Step 2: Synthesis of 3-(3,4-bis(2-methoxyethoxy)phenyl)-1H-1,2,4-triazol-5-amine (intermediate 2):

[0179] A suspension of aminoguanidine nitrate (737.08 mg, 5.38 mmol) and K₂CO₃ (743.02 mg, 5.38 mmol) in N,N-dimethylformamide (DMF) (5 mL) was stirred at room temperature for 1 h. A solution of 3,4-bis(2-methoxyethoxy)benzoic acid (intermediate 1) (1.32 g, 4.89 mmol) in N,N-dimethylformamide (DMF) (10 mL) was treated at 0 °C with N,N'-carbonyldiimidazole (CDI) (871.77 mg, 5.38 mmol). The solution was stirred at room temperature for 1 h. Then it was added to the suspension. The resulting mixture was stirred at room temperature for 3 h, then heated to 100 °C and maintained for 10 h. The mixture was cooled to room temperature, filtered, and concentrated. The residue was analyzed by reversed-phase HPLC (C10). 18 Purification yielded 3-(3,4-bis(2-methoxyethoxy)phenyl)-1H-1,2,4-triazol-5-amine (intermediate 2) as a white solid (800 mg, yield 53.09%). 1 H NMR (400MHz, d6-DMSO) δ11.93(s,1H),7.43-7.40(m,2H),6.99-6.97(m,1H),6.01(s,2H),4.12-4.09(m,4H),3.69-3.64(m,4H),3.33(s,6H).

[0180] Step 3: Synthesis of 3-(3,4-bis(2-methoxyethoxy)phenyl)-N-(2-chloro-6-fluorobenzyl)-1H-1,2,4-triazol-5-amine (34):

[0181] A mixture of 3-(3,4-bis(2-methoxyethoxy)phenyl)-1H-1,2,4-triazol-5-amine (intermediate 2) (800 mg, 2.59 mmol), 2-chloro-6-fluorobenzaldehyde (1.03 g, 6.49 mmol), and p-toluenesulfonic acid (44.7 mg, 0.26 mmol) in isopropanol (10 mL) was heated under reflux overnight. After cooling the solution to room temperature, NaBH3CN (652.2 mg, 10.38 mmol) and acetic acid (1.5 mL) were added. The resulting mixture was then stirred at room temperature for 5 h and quenched with water. The solvent was removed under reduced pressure, and the residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give 3-(3,4-bis(2-methoxyethoxy)phenyl)-N-(2-chloro-6-fluorophenyl)-1H-1,2,4-triazol-5-amine (120 mg, yield 10.26%) as a white solid. Mass spectrometry (ESI) m / z C 21 H 24 ClFN4O4[M+H] + Calculated values: 451.15, 453.15; measured values: 451.44, 453.37. 1 HNMR (400MHz, CDCl3) δ7.46 (s, 1H), 7.45-7.43 (d, J = 8Hz, 1H), 7.25-7.22 (m, 2H), 7.03-7.0 (m, 1H), 6.92-6.90 ( d,J=8Hz,1H),5.18(t,J=6.8Hz,1H),4.65(d,J=6.8Hz,2H),4.19-4.16(m,4H),3.79-3.75(m,4H),3.44(s,6H).

[0182]

[0183] Step 1: Synthesis of 3-methoxy-4-(3-morpholinylpropoxy)benzoic acid (intermediate 1):

[0184] A suspension of methyl 4-hydroxy-3-methoxybenzoate (914.2 mg, 5.02 mmol), TBAI (185.4 mg, 0.50 mmol), and K₂CO₃ (763 mg, 5.52 mmol) in N,N-dimethylformamide (DMF) (10 mL) was heated to 100 °C and maintained for 1 h. After cooling the mixture to 50 °C, 4-(3-chloropropyl)morpholine (903.3 mg, 5.52 mmol) was added. The resulting mixture was stirred at 85 °C for 15 h and then cooled to room temperature. The mixture was filtered, concentrated, and the residue was dissolved in THF (40 mL) / H₂O (10 mL) with KOH (1.09 g, 19.5 mmol). The resulting solution was stirred at room temperature for 5 h and quenched with 1N HCl. Extracted with ethyl acetate (30 mL, 3 times), the organic phase was washed with brine (15 mL), dried on Na2SO4, filtered, and concentrated to give 3-methoxy-4-(3-morpholinylpropoxy)benzoic acid (intermediate 1) (1.30 g, yield 87.72%) as a clear oil, which was used directly in the next step without further purification.

[0185] Step 2: Synthesis of 3-(3-methoxy-4-(3-morpholinylpropoxy)phenyl)-1H-1,2,4-triazol-5-amine (intermediate 2):

[0186] A suspension of aminoguanidine nitrate (1.40 g, 10.21 mmol) and K₂CO₃ (1.41 g, 10.21 mmol) in N,N-dimethylformamide (DMF) (10 mL) was stirred at room temperature for 1 h. A solution of 3-methoxy-4-(3-morpholinylpropoxy)benzoic acid (intermediate 1) (0.90 g, 3.05 mmol) in N,N-dimethylformamide (DMF) (10 mL) was treated at 0 °C with N,N'-carbonyldiimidazole (CDI) (1.10 g, 6.78 mmol). The solution was stirred at room temperature for 1 h. Then it was added to the suspension. The resulting mixture was stirred at room temperature for 3 h, then heated to 100 °C and maintained for 10 h. The mixture was cooled to room temperature, filtered, and concentrated. The residue was analyzed by reversed-phase HPLC (C10-C2 ... 18 Purification yielded 3-(3-methoxy-4-(3-morpholinylpropoxy)phenyl)-1H-1,2,4-triazol-5-amine (intermediate 2) (520 mg, 80% purity) as a white solid, which was used directly in the next step. Mass spectrometry (ESI) m / z C 16 H 23 N5O3[M+H] + The calculated value is 334.18, and the measured value is 334.31.

[0187] Step 3: Synthesis of N-(2-chloro-6-fluorobenzyl)-3-(3-methoxy-4-(3-morpholinylpropoxy)phenyl)-1H-1,2,4-triazol-5-amine (32):

[0188] A mixture of 3-(3-methoxy-4-(3-morpholinylpropoxy)phenyl)-1H-1,2,4-triazol-5-amine (intermediate 2) (480 mg, 1.15 mmol, 80% purity), 2-chloro-6-fluorobenzaldehyde (913.12 mg, 5.76 mmol), and p-toluenesulfonic acid (19.83 mg, 0.12 mmol) in isopropanol (10 mL) was heated under reflux overnight. After cooling the solution to room temperature, NaBH3CN (723.81 mg, 11.52 mmol) and acetic acid (1.5 mL) were added. The resulting mixture was then stirred at room temperature for 5 h and quenched with water. The solvent was removed under reduced pressure, and the residue was purified by column chromatography (silica gel, dichloromethane / methanol = 100:1 to 10:1) to give N-(2-chloro-6-fluorophenyl)-3-(3-methoxy-4-(3-morpholinylpropoxy)phenyl)-1H-1,2,4-triazol-5-amine (85 mg, yield 15.51%) as a white solid. Mass spectrometry (ESI) m / z C 23 H 27 ClFN5O3[M+H] + Calculated values: 476.18, 478.18; measured values: 476.39, 478.27. 1 H NMR(400MHz, CDCl3)δ7.46(s,1H),7.45-7.43(d,J=8.4Hz,1H),7.25-7.22(m,2H),7.03-7.0(m,1H),6.92-6.90(d,J=8.4Hz,1H),5.08(brs ,1H),4.65(dd,J=6.8,1.2Hz,2H),4.15-4.10(m,2H),3.90(s,3H),3.75-3.72(m,4H),2.61-2.58(m,2H),2.54(brs,4H),2.08-2.04(m,2H).

[0189] plasmid construction

[0190] Human p38MAPK and MKK6 were cloned into the pet28a vector using Nco I / Not I, Nde I, and Sal I restriction sites, respectively. The pet28a-hMKK6 was then mutated using a site-directed mutagenesis kit from Transgen to induce the S207D / T211D mutation. The inserted genes were sequenced to ensure sequence accuracy.

[0191] Purification of fusion proteins

[0192] Protein expression was performed using E. coli BL21DE3 and LB medium. Cultures were grown at 37°C until an OD600 nm of 1.2 was reached, then cooled to 18°C ​​with shaking for 1 h, followed by induction at 16°C with 0.2 mM IPTG for 16 h. Protein purification was performed as previously described. Briefly, cells were harvested and purified using Ni resin and a MonoQ HP column. The combined protein peak fractions were then further purified using a Superdex-75 column with 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 5% glycerol, 10 mM MgCl2, and 5 mM DTT as eluents. All purification steps were performed at 4°C.

[0193] P38 MAPK kinase activity assay

[0194] 1000 ng of inactive hMAPK14 was activated with 100 ng of constitutively active hMKK6DD in a 15 μL reaction volume containing 50 μM ATP at 30 °C for 30 min. Kinase activity was assessed using the Z'-LYTE kinase activity assay kit for Ser / Thr 15 peptides (Invitrogen, Carlsbad, CA). The standard reaction for compound screening contained 100 nM hMAPK14, 1 mM peptide substrate, 100 μM ATP, 50 mM MHEPES (pH 7.4), 10 mM MgCl2, 0.01% Brij-35, and 0.5% DMSO.

[0195] Cell culture

[0196] Jurkat T and Ramos B cells were cultured in RPMI 1640 medium supplemented with 10% FBS, 100 μg penicillin and streptomycin per mL.

[0197] Antibody

[0198] The anti-BTK (Tyr223), anti-ZAP-70, anti-PLCγ-2, anti-p-BTK (Y223), anti-p-PLCγ-2 (Y1217), anti-p-ZAP-70 (Y319), and anti-GAPDH used for Western blots were purchased from Cell Signaling Technology (Danvers, MA, USA).

[0199] B and T cell activation and phosphorylation immunoblotting analysis (Phospho-Blots)

[0200] Ramos B cells or Jurkat T cells (2 × 10⁻⁶) were cultured at 37°C in a 5% CO₂ incubator with or without different concentrations of the compound. 6 1.5 h. Then add goat anti-human IgM F(ab')2 (10 μg / mL; Invitrogen) or Dynabeads Human T-activator CD3 / CD28 to stimulate Ramos B cells or Jurkat T cells at 37 °C for 5 min, respectively. Centrifuge the cells, wash once with cold DPBS, and then dissolve them on ice for 20 min in RIPA buffer (Sigma-Aldrich) containing phospholipase inhibitor Cocktail 2 and protease inhibitor Cocktail (both from Sigma-Aldrich). Then centrifuge the sample at 20,000 g for 20 min at 4 °C. Collect the supernatant, dilute it 5-fold with loading buffer, perform SDS-PAGE electrophoresis, and then perform immunoblotting analysis.

[0201] The compound exhibits inhibitory activity against kinase targets associated with some autoimmune diseases.

[0202]

[0203]

[0204]

[0205]

[0206] *NA: Not available.

[0207] We also determined the phosphorylation levels of protein targets in the signaling pathway of Ramos B cells treated with or without the compounds. Ramos B cells treated with or without the compounds for 1.5 h were activated (+) or unactivated (-) with IgMF(ab′)2 for 5 min. SDS-PAGE electrophoresis and Western blotting analysis were performed on the lysates of the harvested cells. After activation of the BCR pathway in Ramos cells with goat anti-human IgMF(ab′)2, exemplary compounds 1, 6, 11, 17, and 20 showed dose-dependent inhibition of BTK autophosphorylation and phosphorylation of the BTK physiological substrate PLCγ. The cellular activity of these compounds was consistent with their performance in enzymatic assays.

[0208] We also determined the phosphorylation levels of protein targets in the signaling pathway of Jurkat T cells treated with or without the compounds. Jurkat T cells treated with or without the compounds for 1.5 h were activated (+) or unactivated (-) for 5 min with Dynabeads Human T-activator CD3 / CD28. Lysates of the harvested cells were subjected to SDS-PAGE electrophoresis and Western blot analysis. After activation of the TCR pathway in Jurkat cells with Dynabeads Human T-activator CD3 / CD28, exemplary compounds 1, 6, 11, 17, and 20 showed a dose-dependent inhibition of phosphorylation of the LCK physiological substrates ZAP-70 and LAT. The cellular activity of these compounds was consistent with their performance in enzymatic assays.

[0209] References

[0210] 1Davidson, A. & Diamond, B. Autoimmune diseases. The New England journal of medicine 345, 340-350 (2001).

[0211] 2Marrack, P., Kappler, J. & Kotzin, BLAutoimmune disease: why and where it occurs. Nat Med 7, 899-905, doi:10.1038 / 90935 90935[pii](2001).

[0212] 3Lanzavecchia, A., Iezzi, G. & Viola, A. From TCR engagement to T cellactivation: a kinetic view of T cell behavior. Cell 96,1-4, doi:S0092-8674(00)80952-6[pii](1999).

[0213] 4Pierce, SK Lipid rafts and B-cell activation. Nat Rev Immunol 2,96-105, doi:10.1038 / nri726(2002).

[0214] 5Flanagan,M.E.et al.Discovery of CP-690,550:a potent and selectiveJanus kinase(JAK)inhibitor for the treatment of autoimmune diseases and organtransplant rejection.J Med Chem 53,8468-8484,doi:10.1021 / jm1004286(2010).

[0215] 6Honigberg,L.A.et al.The Bruton tyrosine kinase inhibitor PCI-32765blocks B-cell activation and is efficacious in models of autoimmune diseaseand B-cell malignancy.Proc Natl Acad Sci U S A 107,13075-13080,doi:10.1073 / pnas.1004594107 1004594107[pii](2010).

[0216] 7Abdel-Magid,A.F.Spleen Tyrosine Kinase Inhibitors(SYK)as PotentialTreatment for Autoimmune and Inflammatory Disorders:Patent Highlight.ACS MedChem Lett 4,18-19,doi:10.1021 / ml300405d(2013).

[0217] 8Bhagwat,S.S.Kinase inhibitors for the treatment of inflammatory andautoimmune disorders.Purinergic Signal 5,107-115,doi:10.1007 / s11302-008-9117-z(2009).

[0218] 9Gaestel,M.,Kotlyarov,A.&Kracht,M.Targeting innate immunity proteinkinase signalling in inflammation.Nat Rev Drug Discov 8,480-499,doi:10.1038 / nrd2829nrd2829[pii](2009).

[0219] 10van der Merwe,P.A.&Dushek,O.Mechanisms for T cell receptortriggering.Nat Rev Immunol 11,47-55,doi:10.1038 / nri2887nri2887[pii](2011).

[0220] 11Palacios,E.H.&Weiss,A.Function of the Src-family kinases,Lck andFyn,in T-cell development and activation.Oncogene 23,7990-8000,doi:1208074[pii]10.1038 / sj.onc.1208074(2004).

[0221] 12Chan,A.C.et al.Activation of ZAP-70 kinase activity byphosphorylation of tyrosine 493 is required for lymphocyte antigen receptorfunction.EMBO J 14,2499-2508(1995).

[0222] 13Wang,H.et al.ZAP-70:an essential kinase in T-cell signaling.ColdSpring Harb Perspect Biol 2,a002279,doi:10.1101 / cshperspect.a002279 2 / 5 / a002279[pii](2010).

[0223] 14Seggewiss,R.et al.Imatinib inhibits T-cell receptor-mediated T-cellproliferation and activation in a dose-dependent manner.Blood 105,2473-2479,doi:2004-07-2527[pii]10.1182 / blood-2004-07-2527(2005).

[0224] 15Stachlewitz,R.F.et al.A-770041,a novel and selective small-moleculeinhibitor of Lck,prevents heart allograft rejection.J Pharmacol Exp Ther 315,36-41,doi:jpet.105.089169[pii]10.1124 / jpet.105.089169(2005).

[0225] 16Burchat,A.et al.Discovery of A-770041,a src-family selective orallyactive lck inhibitor that prevents organ allograft rejection.Bioorg Med ChemLett 16,118-122,doi:S0960-894X(05)01213-8[pii]10.1016 / j.bmcl.2005.09.039(2006).

[0226] 17Won,J.et al.Rosmarinic acid inhibits TCR-induced T cell activationand proliferation in an Lck-dependent manner.Eur J Immunol 33,870-879,doi:10.1002 / eji.200323010(2003).

[0227] 18Youn,J.et al.Beneficial effects of rosmarinic acid on suppressionof collagen induced arthritis.J Rheumatol 30,1203-1207,doi:0315162X-30-1203[pii](2003).

[0228] 19Development of the Potent Anti-Rheumatoid Arthritis CompoundDerived from Rosmarinic Acid and the Evaluation of the Activity in Collagen-Induced Arthritis Mouse Model.248-252(2015).

[0229] 20Abraham,N.,Miceli,M.C.,Parnes,J.R.&Veillette,A.Enhancement of T-cell responsiveness by the lymphocyte-specific tyrosine protein kinasep56lck.Nature 350,62-66,doi:10.1038 / 350062a0(1991).

[0230] 21Cooke,M.P.,Abraham,K.M.,Forbush,K.A.&Perlmutter,R.M.Regulation of Tcell receptor signaling by a src family protein-tyrosine kinase(p59fyn).Cell65,281-291,doi:0092-8674(91)90162-R[pii](1991).

[0231] 22Appleby,M.W.et al.Defective T cell receptor signaling in micelacking the thymic isoform of p59fyn.Cell 70,751-763,doi:0092-8674(92)90309-Z[pii](1992).

[0232] 23Molina,T.J.et al.Profound block in thymocyte development in micelacking p56lck.Nature 357,161-164,doi:10.1038 / 357161a0(1992).

[0233] 24Groves,T.et al.Fyn can partially substitute for Lck in T lymphocytedevelopment.Immunity 5,417-428,doi:S1074-7613(00)80498-7[pii](1996).

[0234] 25Denny,M.F.,Patai,B.&Straus,D.B.Differential T-cell antigen receptorsignaling mediated by the Src family kinases Lck and Fyn.Mol Cell Biol 20,1426-1435(2000).

[0235] 26Webb,Y.,Hermida-Matsumoto,L.&Resh,M.D.Inhibition of proteinpalmitoylation,raft localization,and T cell signaling by 2-bromopalmitate andpolyunsaturated fatty acids.J Biol Chem 275,261-270(2000).

[0236] 27van der Heide,J.J.,Bilo,H.J.,Donker,J.M.,Wilmink,J.M.&Tegzess,A.M.Effect of dietary fish oil on renal function and rejection incyclosporine-treated recipients of renal transplants.The New England journalof medicine 329,769-773,doi:10.1056 / NEJM199309093291105(1993).

[0237] 28Lowenberg,M.et al.Rapid immunosuppressive effects ofglucocorticoids mediated through Lck and Fyn.Blood 106,1703-1710,doi:2004-12-4790[pii]10.1182 / blood-2004-12-4790(2005).

[0238] 29Edwards,J.C.et al.Efficacy of B-cell-targeted therapy withrituximab in patients with rheumatoid arthritis.The New England journal ofmedicine 350,2572-2581,doi:10.1056 / NEJMoa032534350 / 25 / 2572[pii](2004).

[0239] 30Buggy,J.J.&Elias,L.Bruton tyrosine kinase(BTK)and its role in B-cell malignancy.Int Rev Immunol 31,119-132,doi:10.3109 / 08830185.2012.664797(2012).

[0240] 31Rawlings,D.J.et al.Mutation of unique region of Bruton's tyrosinekinase in immunodeficient XID mice.Science 261,358-361(1993).

[0241] 32Lou,Y.et al.Structure-based drug design of RN486,a potent andselective Bruton's tyrosine kinase(BTK)inhibitor,for the treatment ofrheumatoid arthritis.J Med Chem 58,512-516,doi:10.1021 / jm500305p(2015).

[0242] 33Kotlyarov,A.et al.MAPKAP kinase 2 is essential for LPS-induced TNF-alpha biosynthesis.Nat Cell Biol 1,94-97,doi:10.1038 / 10061(1999).

[0243] 34Brook,M.,Sully,G.,Clark,A.R.&Saklatvala,J.Regulation of tumournecrosis factor alpha mRNA stability by the mitogen-activated protein kinasep38 signalling cascade.FEBS Lett 483,57-61,doi:S0014-5793(00)02084-6[pii](2000).

[0244] 35Haddad,J.J.VX-745.Vertex Pharmaceuticals.Curr Opin Investig Drugs2,1070-1076(2001).

[0245] 36Hollenbach,E.et al.Inhibition of p38 MAP kinase-and RICK / NF-kappaB-signaling suppresses inflammatory bowel disease.FASEB J 18,1550-1552,doi:10.1096 / fj.04-1642fje04-1642fje[pii](2004).

[0246] 37Clark,A.R.&Dean,J.L.The p38 MAPK Pathway in Rheumatoid Arthritis:ASideways Look.Open Rheumatol J 6,209-219,doi:10.2174 / 1874312901206010209TORJ-6-209[pii](2012).

Claims

1. Compounds of Formula I: I， in: R2-R4 are H, and R1 and R5 are halogens or methyl groups; R6 is optional and can be replaced: - Selected from thiazole or thiophene; - Selected from phenyl and pyridyl; and The substituents of R6 are selected from: halogen, -OR', -NR'R", -R', -NO2, and the number of substituents is the total number of open valences on R6 from 0; wherein R' and R" are each independently selected from hydrogen and methyl. Or its pharmaceutically acceptable salt.

2. The compound of claim 1, wherein R6 is phenyl, 3-substituted phenyl, 3,4-substituted phenyl or 3,4,5-substituted phenyl.

3. Compounds having the following structures, or pharmaceutically acceptable salts thereof: 。 4. A pharmaceutical composition comprising a therapeutically effective amount of the compound of any one of claims 1-3 in a unit dose form and one or more pharmaceutically acceptable excipients.

5. A composition comprising a therapeutically effective amount of any one of claims 1-3 and various therapeutic agents having therapeutic activity against autoimmune and / or inflammatory diseases or cancer.

6. Use of the compound of any one of claims 1-3 in the preparation of a medicament for treating diseases associated with undesirable kinase activity, wherein the disease is an allergic disease, an autoimmune disease, an inflammatory disease, or cancer.

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