Benzimidazole compounds and uses thereof

Abstract

This invention proposes a novel compound that effectively inhibits IRAK4 activity, which is a compound of Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, its preparation method, and its use in the preparation of drugs.

Description

[0001] Priority information

[0002] This application claims priority and benefits to patent application No. 202110090538.6, filed with the China National Intellectual Property Administration on January 22, 2021, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention belongs to the field of pharmaceutical chemistry. Specifically, this invention relates to benzimidazole compounds, and more specifically, this invention relates to a benzimidazole compound, its preparation method, and its use in the preparation of drugs. Background Technology

[0004] Interleukin-1 receptor-associated kinase 4 (IRAK4) belongs to the IRAK family, which also includes IRAK1, IRAK2, and IRAKM (I-RAK3). The human IRAK4 gene is located in the p11.12 region of the X chromosome and encodes a 52 kDa protein. Like other family members, the IRAK4 protein contains a serine-threonine kinase domain (KD) and a conserved death domain (DD) at the N-terminus. It is a key molecule in the Toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) signaling pathways, participating in the regulation of intracellular signaling cascades and inflammatory responses. TLRs are a family of transmembrane pattern recognition receptors that play a central role in innate immune signaling. The IL-1R family can initiate immune responses in response to stimulation by various IL-1 cytokines.

[0005] Upon stimulation of IL-1R, IL-18R, and most TLRs, myeloid differentiation factor 88 (MyD88) binds to these receptors as an adaptor protein, subsequently recruiting the IRAK4 protein to the TLRs / IL-1R complex. IRAK4 can interact with MyD88 and IRAK2 through a shared death domain, forming the protein complex Myddosome, which activates the kinase activity of IRAK4, leading to phosphorylation of downstream IRAK1 and / or IRAK2. IRAK1 phosphorylation causes a conformational change, prompting it to bind to tumor necrosis factor receptor-associated factor 6 (TRAF6), thereby activating the serine-threonine kinase TAK1, activating downstream NF-κB, and initiating the transcription and expression of pro-inflammatory factors such as IL-1, IL-8, and IL-33, inducing an inflammatory response. Furthermore, the JNK signaling pathway is also activated by TRAF6, regulating the transcription of genes related to inflammation, apoptosis, and cell proliferation. In mouse models where IRAK4 is knocked out, signal transduction and cellular responses of IL-1, IL-18, and most TLR ligands are severely impaired, validating the important role of IRAK4 in IL-1R, IL-18R, and most TLR signaling pathways (Suzuki N, et al. Nature 2002, 416, 750-756.).

[0006] Due to its crucial role in the cytokine signaling network, both abnormally high expression and silencing of IRAK4 can induce abnormalities in the body's immune system. Specifically, abnormally high IRAK4 expression leads to overactivation of the TRL / IL-1R pathway, resulting in the long-term secretion of high levels of pro-inflammatory factors, causing a persistent inflammatory response, and ultimately inducing autoimmune diseases, including rheumatoid arthritis, psoriasis, systemic lupus erythematosus, and multiple sclerosis. Researchers have found that the IL-1 content in the synovial fluid of patients with rheumatoid arthritis is significantly higher than that in healthy individuals (Nouri AM, et al. Clin Exp Immunol 1984; 55(2):295–302.). Further studies have shown that antagonizing TLR4 can reduce IL-1 secretion and has a preventive effect on joint inflammation in mouse models (Abdollahi-Roodsaz S, et al. 2007; 56(9):2957–67.). Activation of TLR7 and TLR9 can stimulate the production of IFNα in pDC cells, which is also considered a potential trigger for systemic lupus erythematosus (Chiang EY, et al. J Immunol 2011; 186(2):1279–88.). Many other autoimmune diseases, including inflammatory bowel disease (Coccia M, et al. J Exp Med 2012; 209(9):1595–609.) and Sjögren's syndrome (Low HZ, et al. Arthritis Res Ther 2011; 13(3):1–7.), are associated with enhanced TLR signaling, further suggesting the potential efficacy of inhibiting IRAK4 in autoimmune diseases.

[0007] IRAK4 has also been reported to play a key role in the development and progression of various malignant tumors, including melanoma, Waldenstrom's macroglobulinemia (WM), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (T-ALL), and ABC-DLCBL. The upregulation of the myddosome complex caused by the MyD88-L256P functional enhancement mutation leads to NF-κB activation, resulting in the survival and proliferation of a series of tumor cell lines. Immunohistochemical results from melanoma biopsies showed that phosphorylated IRAK4 was highly expressed in the lesion tissue (Srivastava R, et al. Cancer Res 2012; 72(23):6209–16.). Increased mRNA expression levels of IRAK1 and IRAK4, as well as elevated phosphorylation levels of IRAK1 and IRAK4, were also observed in T-ALL cells (Li Z, et al. J Clin Invest 2015; 125(3):1081–97.). Silencing IRAK4 with shRNA or inhibiting its enzymatic activity with small molecules weakens cell proliferation in samples from T-ALL patients, indicating that IRAK4 signal transduction is a key factor in disease progression. Furthermore, it has been reported that IRAK4 inhibitors, in combination with Ibrutinib (a BTK inhibitor), synergistically inhibit the proliferation of the MyD88-L256P-mutant ABC-DLBCL cell line (Kelly PN, et al. J ExpMed 2015; 212(13):2189–201.). These findings all confirm the potential value of the IRAK4 target in cancer therapy.

[0008] Furthermore, activation mutations in Fms-like tyrosine kinase (FLT3) lead to its autophosphorylation and the activation of intracellular signaling pathways, thereby promoting the survival and proliferation of leukemia cells and contributing to adaptive resistance to FLT3 inhibitors in AML. Researchers have confirmed that the mechanism by which FLT3 inhibitors induce resistance to FLT3-ITD mutants is related to compensatory activation of IRAK1 / 4. IRAK1 / 4 signaling leads to TLR pathway activation, resulting in autoimmune activation. Knockdown of IRAK4 or inhibition of IRAK4 activity has demonstrated the necessity of IRAK1 / 4 in the process of adaptive resistance in FLT3-ITD mutants (Melgar K, et al. Sci Transl Med 2019; 508(11)), suggesting the potential of simultaneously inhibiting FLT3 signaling and compensatory activation of IRAK1 / 4 in treating FLT3-mutant AML patients. Summary of the Invention

[0009] The present invention aims to propose a novel IRAK4 inhibitor that can be used to prepare drugs for treating tumor-related diseases.

[0010] In a first aspect, the present invention provides a compound, which is a compound of Formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug:

[0011]

[0012] in,

[0013] L represents non-existence or -O-(CH2). m - where m is 0, 1, 2, or 3;

[0014] R 1 Selected from unsubstituted or R a Substituted C1-C6 alkyl groups, or unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups; the R-substituted a Among the substituted C1-C6 alkyl groups, or, as described above, those subjected to R a In the substituted 4-8 membered heterocyclic alkyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when there are multiple substituents, the substituents may be identical or different; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0015] When L is -O-(CH2) m -time,-LR 1 For -O-(CH2) m -R 1 ;

[0016] Ring A is selected from 5-8-membered heteroaryl or 6-10-membered aryl, wherein the heteroatom in the 5-8-membered heteroaryl is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0017] n is 1 or 2;

[0018] R 2 Selected from halogens, hydroxyl groups, cyano groups, amino groups, C1-C6 alkyl groups, C1-C6 alkyl-O- groups, C1-C6 alkyl groups substituted with 1-5 identical or different halogens, unsubstituted groups, or groups substituted with R. b Substituted 5-8 aryl groups, unsubstituted or R bSubstituted 6-10 aryl groups, unsubstituted or R b Substituted 4-8 membered heterocyclic alkyl groups, or unsubstituted or R b Substituted 4-8 membered heterocyclic alkenyl groups; the R-substituted b Substituted 5-8 aryl groups, R b The 6-10 aryl group that was replaced by R b Substituted 4-8 membered heterocyclic alkyl groups, or R b In the substituted 4-8 membered heterocyclic alkenyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when n is not 1, R 2 Independently, they can be either the same or different;

[0019] When ring A is a 5-8 membered heteroaryl group, R 1 Not replaced or by R a Substituted C1-C6 alkyl groups.

[0020] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted C1-C6 alkyl group is used, the C1-C6 alkyl group is a C1-C3 alkyl group, preferably methyl, ethyl, n-propyl, or isopropyl.

[0021] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted 4-8 membered heterocyclic alkyl group is used, the 4-8 membered heterocyclic alkyl group is a nitrogen-containing heterocyclic alkyl group, preferably azacyclobutane, azacyclopentane, azacyclohexane or morpholino.

[0022] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted 4-8 membered heterocyclic alkyl group is used, the 4-8 membered heterocyclic alkyl group is an oxygen-containing heterocyclic alkyl group, preferably an oxetane, an oxecyclopentane, or an oxecyclohexane.

[0023] In a preferred embodiment of the present invention, when R 1 For R a Substituted C1-C6 alkyl groups, or, R a When the substituted 4-8 membered heterocyclic alkyl group is used, the number of substitutions is independently 1-3, preferably 1.

[0024] In a preferred embodiment of the present invention, R a It is a hydroxyl group.

[0025] In a preferred embodiment of the present invention, when R a When the halogen is halogen, the halogen is F, Cl, Br, or I, preferably F or Cl.

[0026] In a preferred embodiment of the present invention, when R a When the alkyl group is C1-C6, the C1-C6 alkyl group is C1-C3 alkyl group, preferably methyl, ethyl, n-propyl, or isopropyl.

[0027] In a preferred embodiment of the present invention, when ring A is selected from 5-8-membered heteroaryl groups, the 5-8-membered heteroaryl group is pyrrole, pyrazole, triazole, furan, oxazole, thiophene, thiazole, pyridine, pyrazine or pyrimidine, preferably oxazole or pyridine.

[0028] In a preferred embodiment of the present invention, when ring A is a 6-10 aryl group, the 6-10 aryl group is phenyl or naphthyl, preferably phenyl.

[0029] In a preferred embodiment of the present invention, when R 2 For not replaced or by R b The substituted 5-8 heteroaryl group is pyrrole, pyrazole, triazole, furan, oxazole, thiophene, thiazole, pyridine, pyrazine or pyrimidine, preferably pyrazole or pyridine.

[0030] In a preferred embodiment of the present invention, when R 2 For not replaced or by R b When the 6-10 aryl group is substituted, the 6-10 aryl group is phenyl or naphthyl, preferably phenyl.

[0031] In a preferred embodiment of the present invention, when R 2 For not replaced or by R b When the substituted 4-8 membered heterocyclic alkyl group is used, the 4-8 membered heterocyclic alkyl group is independently aziridine, oxetane, tetrahydropyrrolidinyl, tetrahydrofuranyl, hexahydropyran or tetrahydro-2H-thiopyran 1,1-dioxide, preferably aziridine or oxetane.

[0032] In a preferred embodiment of the present invention, when R 2 For not replaced or by R bWhen the substituted 4-8 membered heterocyclic alkenyl group is used, the 4-8 membered heterocyclic alkenyl group is independently dihydropyridyl, tetrahydropyridyl, tetrahydropyrimidinyl, pyrrolinyl, imidazolinyl, pyrazolinyl, dihydroimidazolinyl, dihydropyrazolyl, dihydrooxazolyl, dihydrooxadiazolyl, dihydrothiazolinyl, dihydroisothiazolinyl, dihydrothiophenyl, dihydropyrrolyl, 3,4-dihydro-2H-pyranyl, dihydrofuranyl, dihydropyrazinyl, dihydropyrimidinyl or fluorodihydrofuranyl, preferably 1,2,3,4-tetrahydropyridyl, 1,2-dihydropyridyl, 1,4-dihydropyridyl, 1,2,3,6-tetrahydropyridyl, 3,4-dihydro-2H-pyranyl or dihydrofuranyl.

[0033] In a preferred embodiment of the present invention, when R 2 For R b Substituted 5-8 aryl groups, R b When the 6-10 aryl group is replaced, it is R b Substituted 4-8 membered heterocyclic alkyl groups, or R b When the substituted 4-8 membered heterocyclic alkenyl groups are used, the number of substitutions is independently 1-3, preferably 1 or 2.

[0034] In a preferred embodiment of the present invention, when R 2 When the halogen is halogen, the halogen is F, Cl, Br, or I, preferably F or Cl.

[0035] In a preferred embodiment of the present invention, when R 2 When the alkyl group is C1-C6, the C1-C6 alkyl group is C1-C3 alkyl group, preferably methyl, ethyl, n-propyl, or isopropyl.

[0036] In a preferred embodiment of the present invention, when R 2 When the C1-C6 alkyl group is substituted with 1-5 identical or different halogens, the C1-C6 alkyl group is a C1-C3 alkyl group, preferably methyl, ethyl, n-propyl, or isopropyl.

[0037] In a preferred embodiment of the present invention, when R 2 When the halogen is a C1-C6 alkyl group substituted with 1-5 identical or different halogens, the halogen is F, Cl, Br, or I, preferably F or Cl.

[0038] In a preferred embodiment of the present invention, when R 2 When the halogen is a C1-C6 alkyl group substituted with 1 to 5 identical or different halogens, the halogen is 1, 2 or 3.

[0039] In a preferred embodiment of the present invention, when R b When the halogen is halogen, the halogen is F, Cl, Br, or I, preferably F or Cl.

[0040] In a preferred embodiment of the present invention, when R b When the alkyl group is C1-C6, the C1-C6 alkyl group is C1-C3 alkyl group, preferably methyl, ethyl, n-propyl, or isopropyl.

[0041] In a preferred embodiment of the present invention, when R b When the C1-C6 alkyl group is substituted with 1-5 identical or different halogens, the C1-C6 alkyl group is a C1-C3 alkyl group, preferably methyl, ethyl, n-propyl, or isopropyl.

[0042] In a preferred embodiment of the present invention, when R b When the halogen is a C1-C6 alkyl group substituted with 1-5 identical or different halogens, the halogen is F, Cl, Br, or I, preferably F or Cl.

[0043] In a preferred embodiment of the present invention, when R b When the halogen is a C1-C6 alkyl group substituted with 1 to 5 identical or different halogens, the halogen is 1, 2 or 3.

[0044] In a preferred embodiment of the present invention for

[0045] In a preferred embodiment of the present invention for

[0046] In a preferred embodiment of the present invention, the compound represented by Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs are:

[0047]

[0048] L represents non-existence or -O-(CH2). m - where m is 0, 1, 2, or 3;

[0049] R 1 For not replaced or by R a Substituted 4-8 membered heterocyclic alkyl groups; the unsubstituted or R-substituted ones a In the substituted 4-8 membered heterocyclic alkyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when there are multiple substituents, the substituents may be identical or different; the unsubstituted or R-substituted... aIn the substituted 4-8 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0050] When L is -O-(CH2) m -time,-LR 1 For -O-(CH2) m -R 1 ;

[0051] n is 1 or 2;

[0052] R 2 Selected from unsubstituted or R b Substituted 5-8 aryl groups, unsubstituted or R b The substituted 6-10 aryl, halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, or C1-C6 alkyl substituted with 1-5 identical or different halogens; the R-substituted b Substituted 5-8 heteroaryl groups, or, R b In the substituted 6-10 aryl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when n is not 1, R 2 Independently, they can be the same or different.

[0053] In a preferred embodiment of the present invention, L represents the absence of -O-(CH2). m - where m is 0, 1, 2, or 3;

[0054] R 1 For not replaced or by R a Substituted 4-6 membered heterocyclic alkyl groups; the unsubstituted or R-substituted ones a In the substituted 4-6 membered heterocyclic alkyl groups, each substitution independently refers to one or more hydroxyl groups substituted; the unsubstituted or R-substituted groups... a In the substituted 4-6 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0055] n is 1 or 2;

[0056] R 2 Selected from unsubstituted or R b Substituted 5- or 6-membered heteroaryl groups, the unsubstituted or R-substituted groups b In the substituted 5- or 6-membered heteroaryl groups, each substitution independently refers to one or more C1-C3 alkyl substitutions; when n is not 1, R 2Independently identical or different; the unsubstituted or R b In the substituted 5- or 6-membered heteroaryl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1 to 3.

[0057] In a preferred embodiment of the present invention, the compound represented by Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs are:

[0058]

[0059] L represents non-existence or -O-(CH2). m - where m is 0, 1, 2, or 3;

[0060] R 1 Selected from unsubstituted or R a Substituted C1-C6 alkyl groups, or unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups; the R-substituted a Among the substituted C1-C6 alkyl groups, or, as described above, those subjected to R a In the substituted 4-8 membered heterocyclic alkyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when there are multiple substituents, the substituents may be identical or different; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0061] When L is -O-(CH2) m -time,-LR 1 For -O-(CH2) m -R 1 ;

[0062] n is 1 or 2;

[0063] R 2 Selected from halogens, hydroxyl groups, cyano groups, amino groups, C1-C6 alkyl groups, C1-C6 alkyl-O- groups, C1-C6 alkyl groups substituted with 1-5 identical or different halogens, unsubstituted groups, or groups substituted with R. b Substituted 5-8 aryl groups, unsubstituted or R b Substituted 6-10 aryl groups, unsubstituted or R b Substituted 4-8 membered heterocyclic alkyl groups, or unsubstituted or R b Substituted 4-8 membered heterocyclic alkenyl groups; the R-substituted bSubstituted 5-8 aryl groups, R b The 6-10 aryl group that was replaced by R b Substituted 4-8 membered heterocyclic alkyl groups, or R b In the substituted 4-8 membered heterocyclic alkenyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when n is not 1, R 2 Independently, they can be the same or different.

[0064] In a preferred embodiment of the present invention, L represents the absence of -O-(CH2). m - where m is 0, 1, 2, or 3;

[0065] R 1 Selected from unsubstituted or R a Substituted C1-C4 alkyl groups, wherein the R a In the substituted C1-C4 alkyl groups, each substitution independently refers to one or more C1-C3 alkyl or hydroxyl substituents from the following substituents, and when there are multiple substituents, the substituents may be the same or different;

[0066] n is 1 or 2;

[0067] R 2 Selected from halogens, C1-C3 alkyl groups, and C1-C3 alkyl groups substituted with 1-5 identical or different halogens, where n is not 1, R 2 Independently, they can be the same or different.

[0068] In a preferred embodiment of the present invention, the compound represented by Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs are:

[0069]

[0070] L represents non-existence or -O-(CH2). m - where m is 0, 1, 2, or 3;

[0071] R 1 For not replaced or by R a Substituted 4-8 membered heterocyclic alkyl groups; the R-substituted aIn the substituted 4-8 membered heterocyclic alkyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when there are multiple substituents, the substituents may be identical or different; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0072] When L is -O-(CH2) m -time,-LR 1 For -O-(CH2) m -R 1 ;

[0073] n is 1 or 2;

[0074] R 2 Selected from unsubstituted or R b Substituted 5-8 aryl groups, unsubstituted or R b The substituted 6-10 aryl, halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, or C1-C6 alkyl substituted with 1-5 identical or different halogens; the R-substituted b Substituted 5-8 heteroaryl groups, or, R b In the substituted 6-10 aryl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkyl-O-, -COOH, -C(=O)NH2, C1-C6 alkyl substituted with 1-5 identical or different halogens; when n is not 1, R 2 Independently, they can be the same or different.

[0075] In a preferred embodiment of the present invention, L represents the absence of -O-(CH2). m - where m is 0, 1, 2, or 3;

[0076] R 1 For not replaced or by R a Substituted 4-6 membered heterocyclic alkyl groups, said to be R a In the substituted 4-6 membered heterocyclic alkyl groups, each substitution independently refers to one or more hydroxyl groups being substituted; when there are multiple substituents, the substituents may be the same or different; the unsubstituted or R-substituted... a In the substituted 4-6 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0077] n is 1 or 2;

[0078] R 2 Selected from unsubstituted or R b Substituted 5- or 6-membered heteroaryl groups, or C1-C3 alkyl groups substituted with 1-4 identical or different halogens; the R-substituted... b In the substituted 5- or 6-membered heteroaryl groups, each substitution independently refers to one or more C1-C3 alkyl substitutions, where R is not 1. 2 Independently identical or different, the unsubstituted or R b In the substituted 5- or 6-membered heteroaryl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1 to 3.

[0079] In a preferred embodiment of the present invention, ring A is a 5- or 6-membered heteroaryl group, wherein the heteroatom is selected from one or more of N, O, S and P, and the number of heteroatoms is 1-3.

[0080] R 2 Selected from 1-3 C1-C3 alkyl groups substituted with the same or different halogens, or those modified by R a The substituted 5- or 6-membered heteroaryl group, said to be R a In the substituted 5- or 6-membered heteroaryl group, each substitution independently refers to one or more C1-C3 alkyl substitutions, and in the 5- or 6-membered heteroaryl group, the heteroatom is selected from N, O, S or P, and the number of heteroatoms is 1.

[0081] When L does not exist, R 1 Selected from unsubstituted or R b The substituted 5- or 6-membered heterocyclic group, said to be R b In the substituted 5- or 6-membered heterocyclic group, each substitution independently refers to one or more hydroxyl substitutions, and in the 5- or 6-membered heterocyclic group, the heteroatom is selected from one or more of N, O, S and P, and the number of heteroatoms is 1 to 3;

[0082] When L is -O-(CH2) m - At that time, R 1 The heteroatom is selected from a 4- or 5-membered heterocyclic group, wherein the heteroatom is selected from one or more of N, O, S and P, and the number of heteroatoms is 1 to 3.

[0083] In a preferred embodiment of the present invention, the compound as described in Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs are selected from any of the following compounds:

[0084]

[0085]

[0086] In a second aspect, the present invention provides a pharmaceutical composition comprising the compound shown in Formula I above, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and pharmaceutically acceptable excipients.

[0087] According to specific embodiments of the present invention, the pharmaceutical compositions of the present invention, comprising a therapeutically effective dose of the aforementioned compounds, their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and pharmaceutically acceptable pharmaceutical carriers, diluents, or excipients, can be mixed to prepare a pharmaceutical formulation suitable for oral or parenteral administration. Administration methods include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulation can be administered via any route, such as by infusion or bolus, or by absorption through the epithelium or mucous membranes (e.g., oral mucosa or rectum). Administration can be systemic or local. Examples of orally administered formulations include solid or liquid dosage forms, specifically including tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The formulation can be prepared by methods known in the art and contains carriers, diluents, or excipients conventionally used in the field of pharmaceutical formulations.

[0088] In a third aspect, the present invention provides the use of the compounds shown in Formula I, their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions thereof, in the preparation of a medicament for treating diseases related to IRAK4.

[0089] According to specific embodiments of the present invention, the use of the above-described compound or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs or the above-described pharmaceutical compositions in the preparation of a medicament for treating IRAK4-related diseases, said medicament for treating autoimmune diseases and cancers. These autoimmune diseases include, for example, multiple sclerosis, systemic lupus erythematosus, psoriasis, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis, reactive arthritis, systemic juvenile idiopathic arthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, and allergic eczema; these cancers include, for example, brain cancer, kidney cancer, liver cancer, stomach cancer, vaginal cancer, ovarian cancer, gastric tumors, breast cancer, cystocolic cancer, prostate cancer, pancreatic cancer, lung cancer, cervical cancer, testicular cancer, skin cancer, bone cancer, or thyroid cancer; sarcoma, glioblastoma, neuroblastoma, multiple myeloma, gastrointestinal cancer, neck and head tumors, adenomas, adenocarcinomas, keratoacanthomas, epidermal cancers. Hematologic malignancies include: follicular carcinoma, large cell carcinoma, non-small cell lung cancer, Hodgkin's and non-Hodgkin's lymphoma, breast cancer, follicular carcinoma, papillary carcinoma, seminoma, melanoma; acute myeloid leukemia, chronic myeloid leukemia, diffuse large B-cell lymphoma, activated B-cell-like diffuse large B-cell lymphoma, chronic lymphocytic leukemia, chronic lymphocytic lymphoma, primary exudative lymphoma, Burkitt lymphoma / leukemia, acute lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, Waldenström macroglobulinemia, splenic marginal zone lymphoma, intravascular large B-cell lymphoma, plasmacytoma, and multiple myeloma.

[0090] According to a specific embodiment of the present invention, the compound represented by Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or pharmaceutical compositions are used to inhibit IRAK4 kinase activity.

[0091] The compound shown in Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or pharmaceutical compositions are used to inhibit TNF-α activity.

[0092] The compound, tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or pharmaceutical composition thereof, as shown in Formula I, is used to inhibit the activity of FLT3-WT, FLT3-ITD, or FLT3-D35Y kinases.

[0093] The compound, tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or pharmaceutical composition shown in Formula I is used to inhibit tumor cell proliferation, preferably, the tumor cells are selected from human B-cell lymphoma cells.

[0094] Terms and Definitions

[0095] Unless otherwise stated, the terms and definitions used in this application, including those set forth in the specification and claims, are as follows.

[0096] Those skilled in the art will understand that, according to the conventions used in the art, in the structural formula of this application, Used to describe chemical bonds, which are points where a portion or a substituent is connected to a core or skeletal structure.

[0097] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0098] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable, non-toxic salt of an acid or base, including salts of inorganic acids and bases, and salts of organic acids and bases.

[0099] In addition to pharmaceutically acceptable salts, the present invention also contemplates other salts. These may serve as intermediates in the purification of compounds or in the preparation of other pharmaceutically acceptable salts, or may be used for the identification, characterization, or purification of the compounds of the present invention.

[0100] The term "pharmaceutical composition" refers to a mixture of one or more compounds described herein, or physiologically / pharmaceutical acceptable salts or prodrugs thereof, with other chemical components, such as physiologically / pharmaceutical acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compound to a living organism.

[0101] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of the term "excipient" include, without limitation, binders, disintegrants, lubricants, flow aids, stabilizers, fillers, and diluents. Excipients enhance the handling properties of pharmaceutical formulations, i.e., by increasing flowability and / or adhesion, making the formulation more suitable for direct compression.

[0102] The term "prodrug" refers to a compound of the present invention that can be converted into a biologically active form under physiological conditions or by solvation. The prodrugs of the present invention are prepared by modifying functional groups in the compound; this modification can be performed conventionally or removed in vivo to obtain the parent compound. Prodrugs comprise compounds formed by attaching a hydroxyl or amino group to any group within the compound of the present invention. When a prodrug of the compound of the present invention is administered to a mammalian individual, the prodrug is cleaved to form a free hydroxyl group and a free amino group.

[0103] The term "stereoisomer" refers to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, non-corresponding isomers, and conformational isomers.

[0104] Depending on the choice of raw materials and methods, the compounds of the present invention may exist as one or a mixture of possible isomers, for example as purely optical isomers, or as mixtures of isomers, such as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or multiple chiral centers) in the molecule. The prefixes D and L or (+) and (–) are symbols used to specify the plane-polarized rotation of light induced by the compound, where (–) or L indicates that the compound is levorotatory. Compounds with the prefix (+) or D are dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be called enantiomers, and mixtures of said isomers are generally referred to as mixtures of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or method. Many geometric isomers of alkenes, C=N double bonds, etc., can also exist in the compounds described herein, and all such stable isomers are considered in this invention. When the compounds described herein contain an alkene double bond, unless otherwise stated, such double bond includes E and Z geometric isomers. If the compound contains a disubstituted cycloalkyl group, the substituent of the cycloalkyl group may be in cis or trans (cis- or trans-) configuration.

[0105] When the bonds of the chiral carbon in the formulas of this invention are depicted as straight lines, it should be understood that both the (R) and (S) configurations of the chiral carbon and the resulting enantiomerically pure compounds and mixtures thereof are included within the scope of the general formula. The illustration of racemic or enantiomerically pure compounds in this document is derived from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise stated, wedge-shaped and dashed bonds represent the absolute configuration of a stereocenter.

[0106] Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral formulations, or resolved using conventional techniques. Compounds of the present invention containing asymmetrically substituted carbon atoms can be separated in either an optically active or racemic form. Resolution of racemic mixtures of compounds can be performed by any of many methods known in the art. Exemplary methods include fractional recrystallization using a chiral resolving acid, which is an optically active salt-forming organic acid. Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids such as the D and L forms of β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization methods include stereoisomerically pure α-methylbenzylamine (e.g., S and R forms or diastereoisomeric forms), 2-phenylglycine, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, etc. Resolution of racemic mixtures can also be achieved by elution onto a chromatographic column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). High-performance liquid chromatography (HPLC) or supercritical fluid chromatography (SFC) can be used. The specific method, elution conditions, and column selection can be chosen by those skilled in the art based on the structure of the compound and experimental results. Furthermore, any enantiomer or diastereomeric form of the compound described in this invention can be obtained through stereoorganic synthesis using optically pure starting materials or reagents with known configurations.

[0107] The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom between two positions within a molecule. The compounds of this invention can exhibit tautomerism. Tautomers can exist in two or more interconvertible forms. Proton-transfer tautomers arise from the migration of covalently bonded hydrogen atoms between two atoms. Tautomers generally exist in equilibrium form; attempts to isolate a single tautomer typically yield a mixture whose physicochemical properties are consistent with those of the mixture of compounds. The equilibrium position depends on the intramolecular chemical characteristics. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone form is dominant; while in phenols, the enol form is dominant. This invention encompasses all tautomeric forms of the compounds.

[0108] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium. 2 H), tritium ( 3 H), Iodine-125 125 I) or C-14 14 C). All isotopic variations of the compounds of the present invention, regardless of radioactivity, are included within the scope of the present invention.

[0109] For pharmaceuticals or pharmacologically active agents, the term "effective amount" or "therapeutic effective amount" refers to a sufficient quantity of a drug or agent that is non-toxic but achieves the desired effect. For the oral dosage forms of this invention, the "effective amount" of one active substance in the composition refers to the quantity required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. A suitable effective amount in any given case can be determined by a person skilled in the art through routine testing.

[0110] The terms “active ingredient,” “therapeutic agent,” “active substance,” or “active agent” refer to a chemical entity that can effectively treat a target disorder, disease, or symptom.

[0111] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, including deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is a ketone group (i.e., =O), it means that two hydrogen atoms are replaced. Ketone substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents can be arbitrary on a chemically feasible basis.

[0112] The term "C1-C6 alkyl" should be understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers. In particular, the group has 1, 2 or 3 carbon atoms (“C1-C3 alkyl”), such as methyl, ethyl, n-propyl or isopropyl.

[0113] The term "C1-C6 alkyl-O-" should be understood as an alkyl group connected to the rest of the molecule via an oxygen atom, where "C1-C6 alkyl" has the above definition. Examples include methyl-O- and ethyl-O-.

[0114] The term "6-10 aryl" should be understood as a monocyclic, bicyclic, or tricyclic hydrocarbon ring having 6-10 carbon atoms, exhibiting monovalent aromatic or partially aromatic properties, particularly a ring having 6 carbon atoms ("C6 aryl"), such as a phenyl group; when the 6-10 aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on its substitution site; for example, it can be ortho, para, or meta substituted.

[0115] The term "5-8-membered heteroaryl" should be understood as a monovalent monocyclic, bicyclic, or tricyclic aromatic ring group having 5-8 ring atoms, particularly 5 or 6 carbon atoms, and comprising 1-5 heteroatoms independently selected from N, O, and S. Preferably, it comprises 1-3 monovalent monocyclic, bicyclic, or tricyclic aromatic ring groups independently selected from N, O, and S, and in each case, it may be benzofused. Specifically, the heteroaryl group is selected from thiophene, furanyl, pyrrole, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl, etc.; or pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc.; or borazinyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphridinyl, pteridinyl, carbazolyl, acridineyl, phenazinyl, phenothiazinyl, phenothiazinyl, phenothiazinyl, etc.

[0116] The terms “4-8 membered heterocyclic group” or “4-8 membered heterocyclic alkyl group” should be understood to mean a saturated, unsaturated, or partially saturated monocyclic, bicyclic, or tricyclic ring having 4 to 8 atoms, wherein 1, 2, 3, 4, or 5 ring atoms are selected from N, O, and S, and unless otherwise specified, they may be linked by carbon or nitrogen, wherein -CH 2- The group may optionally be replaced by -C(O)-; and unless otherwise stated to the contrary, the cyclic nitrogen atom or cyclic sulfur atom may optionally be oxidized to form an N-oxide or S-oxide, or the cyclic nitrogen atom may optionally be quaternized; wherein the -NH in the ring may optionally be replaced by an acetyl, formyl, methyl, or methanesulfonyl group; and the ring may optionally be replaced by one or more halogens. It should be understood that when the total number of S and O atoms in the heterocyclic group exceeds 1, these heteroatoms are not adjacent to each other. If the heterocyclic group is bicyclic or tricyclic, at least one ring may optionally be a heteroaromatic ring or an aromatic ring, provided that at least one ring is non-heteroaromatic. If the heterocyclic group is monocyclic, it is necessarily not aromatic. Examples of heterocyclic groups include, but are not limited to, piperidinyl, N-acetylpiperidinyl, N-methylpiperidinyl, N-formylpiperazinyl, N-methanesulfonylpiperazinyl, homopiperazinyl, piperazinyl, azacyclic butyl, oxacyclic butyl, morpholinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, dihydroindolyl, tetrahydropyranyl, dihydro-2H-pyranyl, tetrahydrofuranyl, tetrahydrothiaranyl, tetrahydrothiaran-1-oxide, tetrahydrothiaran-1,1-dioxide, 1H-pyridin-2-one, and 2,5-dioxoimidazolyl.

[0117] The terms "halogen" or "halogen" refer to fluorine, chlorine, bromine, and iodine.

[0118] Furthermore, it should be noted that, unless otherwise explicitly stated, the descriptive phrase "...independently" used in this invention should be interpreted broadly, meaning that the described individuals are independent of each other and can independently be the same or different specific groups. More specifically, the descriptive phrase "...independently" can mean either that the specific options expressed by the same symbols in different groups do not affect each other, or that the specific options expressed by the same symbols in the same group do not affect each other.

[0119] Beneficial effects

[0120] According to embodiments of the present invention, the present invention provides an IRAK4 inhibitor with novel structure, excellent pharmacokinetic properties, and good pharmacodynamics or drug-likeness, which can be used to effectively treat IRAK4-related diseases and symptoms.

[0121] Pharmacokinetic studies in mice, rats, and dogs showed that the compounds of this invention exhibit excellent pharmacokinetic properties, high exposure levels, and good drug-likeness.

[0122] The compounds of this invention all exhibit good inhibitory effects on IRAK4 kinase, especially compounds I-1, I-2, I-3, I-4, I-5, and I-6, which show significant inhibitory effects on IRAK4 kinase. Secondly, the compounds of this invention all exhibit good inhibitory activity against TNF-α production, especially compounds I-1, I-2, and I-3, which show significant inhibitory activity against TNF-α production. In addition, the compounds of this invention all exhibit good inhibitory effects against IL-6 production, indicating that the compounds of this invention have good anti-inflammatory effects.

[0123] The compounds of this invention all exhibit strong inhibitory activity against the proliferation of OCI-LY10 cells, especially compounds I-2 and I-5, which show significant inhibitory activity against the proliferation of OCI-LY10 cells. In addition, compounds I-1 and I-2 also show significant inhibitory effects against wild-type, ITD mutant, and D835Y mutant FLT3 kinases, indicating that the compounds of this invention have good anti-tumor effects.

[0124] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Detailed Implementation

[0125] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0126] Unless otherwise specified, the structures of the compounds in this invention were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts are in units of 10⁻¹⁰. -6 (ppm). The solvents used for NMR determination were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard was tetramethylsilane (TMS).

[0127] The abbreviations of this invention are defined as follows:

[0128] M: Molar concentration, such as 1M hydrochloric acid, which represents a 1 mol / L hydrochloric acid solution.

[0129] N: Equivalent concentration, for example, 2N hydrochloric acid represents a 2 mol / L hydrochloric acid solution.

[0130] DAST: Diethylaminosulfuric acid

[0131] DCM: Dichloromethane

[0132] DEAD: Diethyl azodicarbonate

[0133] DMF: N,N-dimethylformamide

[0134] DMP: Dys-Martin Oxidant

[0135] DMSO: Dimethyl sulfoxide

[0136] DIPEA: Also written as DIEA, diisopropylethylamine, i.e., N,N-diisopropylethylamine.

[0137] EA: Ethyl acetate

[0138] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N,N,-tetramethylurea hexafluorophosphine salt

[0139] PPTS: Pyridine salt of 4-methylbenzenesulfonate

[0140] Pd2(dba)3: Tris(dibenzylacetone)dipalladium

[0141] Ruphos-G3: Mesylate (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II)

[0142] THF: Tetrahydrofuran

[0143] Xantphos: 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene

[0144] Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

[0145] TLC: Thin-layer chromatography

[0146] LC-MS: Liquid chromatography-mass spectrometry

[0147] IC 50 The half-maximum inhibitory concentration (MCI) is the concentration at which half of the maximum inhibitory effect is achieved.

[0148] Preparation 1: Preparation of Intermediate A

[0149] 2-(2-methylpyridin-4-yl)oxazol-4-carboxylic acid

[0150]

[0151] The synthetic route for target intermediate A is as follows:

[0152]

[0153] Step 1: Synthesis of ethyl 2-(2-methylpyridin-4-yl)oxazol-4-carboxylate (A-3)

[0154]

[0155] At room temperature, 3.0 g (21.9 mmol) of 2-methylpyridin-4-yl)boric acid was added to 10 mL of DMF. Under nitrogen protection, ethyl 2-bromooxazol-4-carboxylate (3.0 g, 13.6 mmol) and Pd(dppf)Cl2 (2.2 g, 2.7 mmol) were added, along with potassium carbonate (3.8 g, 11.7 mmol). The mixture was heated to 150 °C and stirred for 16 h. After cooling to room temperature, 100 mL of water was added, and the mixture was extracted with EA (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give 1.13 g (35.7%) of ethyl 2-(2-methylpyridin-4-yl)oxazol-4-carboxylate as a white solid.

[0156] LC-MS, M / Z (ESI): 233.2 [M+H] +

[0157] Step 2: Synthesis of 2-(2-methylpyridin-4-yl)oxazol-4-carboxylic acid (A)

[0158]

[0159] Ethyl 2-(2-methylpyridin-4-yl)oxazol-4-carboxylic acid (1.13 g, 0.43 mmol) was added to THF (10 mL) at room temperature, followed by water (5 mL), methanol (5 mL), and lithium hydroxide monohydrate (585 mg, 24.3 mmol). The mixture was stirred for 18 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give the title compound, 2-(2-methylpyridin-4-yl)oxazol-4-carboxylic acid (1.0 g, 100% yield), as a white solid.

[0160] LC-MS, M / Z (ESI): 205.2 [M+H] + .

[0161] Example 1: Preparation of target compound I-1

[0162] N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholin-1-hydro-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (target compound I-1)

[0163]

[0164] The synthetic route for target compound I-1 is as follows:

[0165]

[0166] Step 1: Synthesis of 4-chloro-N-methyl-2-nitroaniline (1A)

[0167]

[0168] Under ice bath conditions, 50.0 g of 4-chloro-1-fluoro-2-nitrobenzene (0.286 mol) was slowly added to a 250 ml solution of methylamine. The ice bath was then removed, and the mixture was allowed to react at room temperature for 1 h. After the reaction was confirmed to be complete by TLC, the solution was filtered off, and the crude product was obtained by vacuum distillation. The crude product was used directly in the next step of the reaction without purification.

[0169] Step 2: 4-Chloro-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (1B)

[0170]

[0171] The crude 4-chloro-N-methyl-2-nitroaniline obtained in the first step was dissolved in 150 mL of ethanol along with zinc powder (27.0 g, 0.415 mol) and ammonium chloride (36.0 g, 0.679 mol). The mixture was heated to 50 °C and reacted for 2 h. After the reaction was confirmed to be complete by TLC, the mixture was filtered, and the solution was concentrated under reduced pressure to remove the ethanol. The residue was diluted with water (200 mL) and extracted three times with DCM (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the target compound 4-chloro-N-methyl-2-nitroaniline. 1 -Methylphenyl-1,2-diamine (21.0 g, two-step yield 94.3%).

[0172] LC-MS, M / Z (ESI): 157.1 [M+H] +

[0173] Step 3: Synthesis of methyl 3-(5-chloro-1-methyl-1-hydro-benzo[d]imidazol-2-yl)propionate (1C)

[0174]

[0175] 4-chloro-N 1 1,2-methylphenyl-1,2-diamine (9.0 g, 0.058 mol) and methyl 4-oxobutyrate (9.0 g, 0.063 mol) were dissolved in 90 mL of acetic acid, heated to 70 °C and reacted overnight. After the reaction was complete as detected by TLC, the mixture was distilled under reduced pressure. The residue was diluted with water (200 mL) and extracted three times with DCM (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by column chromatography to give the target compound methyl 3-(5-chloro-1-methyl-1-hydro-benzo[d]imidazol-2-yl)propionate (7.0 g, yield 48.3%).

[0176] LC-MS, M / Z (ESI): 253.2 [M+H] +

[0177] Step 4: Synthesis of methyl 3-(5-chloro-1-methyl-6-nitro-1-hydro-benzo[d]imidazol-2-yl)propionate (1D)

[0178]

[0179] Methyl 3-(5-chloro-1-methyl-1-hydro-benzo[d]imidazol-2-yl)propionate (2.0 g, 7.94 mmol) was dissolved in 16 mL of concentrated sulfuric acid under ice bath conditions. Then, a mixture of concentrated nitric acid and concentrated sulfuric acid (HNO3 / H2SO4 = 1:1.4) was slowly added dropwise. The mixture was then heated to room temperature and reacted for 3 h. After the reaction was confirmed to be complete by TLC, saturated NaHCO3 solution was added to adjust the pH to neutral. The mixture was then extracted three times with DCM (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by column chromatography to obtain the target compound methyl 3-(5-chloro-1-methyl-6-nitro-1-hydro-benzo[d]imidazol-2-yl)propionate (1.9 g, yield 80.5%).

[0180] LC-MS, M / Z (ESI): 298.2 [M+H] +

[0181] Step 5: Synthesis of methyl 3-(1-methyl-5-morpholino-6-nitro-1-hydro-benzo[d]imidazol-2-yl)propionate (1E)

[0182]

[0183] Methyl 3-(5-chloro-1-methyl-6-nitro-1-hydro-benzo[d]imidazol-2-yl)propionate (1.09 g, 3.67 mmol), morpholine (1.28 g, 14.7 mmol), chloro(2-dicyclohexylphosphino-2',6'-di-isopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) (566 mg, 0.73 mmol) and cesium carbonate (2.3 g, 7.08 mmol) were dissolved in 11 mL of water. In anhydrous 1,4-dioxane, the air in the reaction flask was purged with nitrogen and then sealed. The reaction was carried out at 110°C for 4 hours. After the reaction was confirmed to be complete by TLC, water (30 mL) was added for dilution. The mixture was extracted three times with DCM (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by column chromatography to obtain the target compound methyl 3-(1-methyl-5-morpholino-6-nitro-1-hydro-benzo[d]imidazol-2-yl)propionate (820 mg, yield 64.1%).

[0184] LC-MS, M / Z (ESI): 349.3 [M+H] +

[0185] Step 6: Synthesis of methyl 3-(6-amino-1-methyl-5-morpholino-1-hydro-benzo[d]imidazol-2-yl)propionate (1F)

[0186]

[0187] Methyl 3-(1-methyl-5-morpholino-6-nitro-1-hydro-benzo[d]imidazol-2-yl)propionate (820 mg, 2.36 mmol), iron powder (526 mg, 9.40 mmol), and ammonium chloride (498 mg, 9.40 mmol) were added to a mixed solvent of ethanol / water = 4 ml / 4 ml. The mixture was refluxed at 90 °C for 2 h. After the reaction was confirmed to be complete by TLC, the mixture was filtered, and the liquid phase was diluted with water (30 mL). The mixture was extracted three times with DCM (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by column chromatography to obtain the target compound methyl 3-(6-amino-1-methyl-5-morpholino-1-hydro-benzo[d]imidazol-2-yl)propionate (480 mg, yield 64.1%).

[0188] LC-MS, M / Z (ESI): 319.3 [M+H] +

[0189] Step 7: Synthesis of methyl 3-(1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamido)-5-morpholino-1-hydro-benzo[d]imidazol-2-yl)propionate (1G)

[0190]

[0191] Methyl 3-(6-amino-1-methyl-5-morpholino-1-hydro-benzo[d]imidazol-2-yl)propionate (240 mg, 0.75 mmol), 2-(2-methylpyridin-4-yl)oxazol-4-carboxylic acid (168 mg, 0.82 mmol), HATU (428 mg, 1.13 mmol), and triethylamine (228 mg, 2.25 mmol) were dissolved in 2.5 ml. In DMF, the reaction was carried out at room temperature for 6 hours. After the reaction was confirmed to be complete by TLC, DMF was removed by vacuum distillation. The residue was diluted with water (30 mL) and then mixed with DCM (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by column chromatography to obtain the target compound methyl 3-(1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamido)-5-morpholino-1-hydro-benzo[d]imidazol-2-yl)propionate (310 mg, yield 81.6%).

[0192] LC-MS, M / Z (ESI): 505.3 [M+H] +

[0193] Step 8: Synthesis of N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholin-1-hydro-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (I-1)

[0194]

[0195] Under ice bath conditions, methyl 3-(1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamido)-5-morpholino-1-hydro-benzo[d]imidazol-2-yl)propionate (50 mg, 0.099 mmol) was dissolved in 1 mL of anhydrous THF. Magnesium methyl bromide (3 mol / L 2-methyltetrahydrofuran solution) (0.25 mL, 0.75 mmol) was slowly added, and the mixture was then allowed to react overnight at room temperature. After TLC analysis to confirm complete reaction, saturated magnesium bromide was added. The mixture was quenched with ammonium chloride solution, diluted with water (30 mL), and then combined with DCM (10 mL × 3). The organic phases were dried over anhydrous sodium sulfate, and the mixture was distilled under reduced pressure. The residue was separated and purified using silica gel plates (DCM:MeOH = 10:1) to obtain the target compound N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholin-1-hydro-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (1.2 mg, yield 2.4%).

[0196] 1 H NMR (400MHz, CDCl3) δ10.61(s,1H),8.73(d,J=4.0Hz,1H),8.60(s,1H),8.40(s,1H),7.83(s,1H),7.76(d,J=4.0Hz,1H), 7.57(s,1H),4.07(t,J=4.0Hz,4H),3.76(s,3H),3.03(t,J=6.0Hz,4H),2.69(s,3H),2.12(t,J=8.0Hz,2H),1.33(s,6H).

[0197] LC-MS, M / Z (ESI): 505.3 [M+H] + .

[0198] Example 2: Preparation of target compound I-2

[0199] (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidin-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide

[0200]

[0201] The synthetic route for the target product I-2 is shown below:

[0202]

[0203] Step 1: Synthesis of 4-bromo-N-methyl-2-nitroaniline (2B)

[0204]

[0205] At room temperature, 10.0 g (45.4 mmol) of 4-bromo-1-fluoro-2-nitrobenzene was added to 100 mL of THF, followed by 20 mL of ethanol solution of methylamine (29%). The mixture was heated to 50 °C and stirred for 16 h. After cooling to room temperature, 500 mL of water was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 8:1) to give crude 4-bromo-N-methyl-2-nitrobenzene (2B) (10.5 g, 100% yield) as a pale yellow liquid.

[0206] LC-MS, M / Z (ESI): 231.2 [M+H] +

[0207] Step 2: 4-Bromo-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (2C)

[0208]

[0209] At room temperature, 10.5 g (45.4 mmol) of 4-bromo-N-methyl-2-nitroaniline was added to 100 mL of THF, followed by 100 mL of ethanol and 50 mL of water. Iron powder (10.5 g, 187.5 mmol) and ammonium chloride (10.5 g, 196.2 mmol) were also added. The mixture was heated to 90 °C and stirred for 3 h. After cooling to room temperature, the mixture was extracted with DCM (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow liquid, 4-bromo-N-methyl-2-nitroaniline. 1 -Methylphenyl-1,2-diamine (2C) (8.8 g, yield 96.3%).

[0210] LC-MS, M / Z (ESI): 201.2 [M+H] +

[0211] Step 3: Synthesis of methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (2D)

[0212]

[0213] The raw material 4-bromo-N was prepared at room temperature. 11,2-methylphenyl-1,2-diamine (8.8 g, 43.7 mmol) was added to 100 mL of DCM and cooled to 0 °C. Methyl 4-chloro-4-oxobutyrate (7.3 g, 48.5 mmol) and triethylamine (13.2 g, 131 mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (500 mL) was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a colorless crude liquid of methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (2D) (9.2 g, yield 66.7%).

[0214] LC-MS, M / Z (ESI): 315.2 [M+H] +

[0215] Step 4: Synthesis of methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (2E)

[0216]

[0217] Methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (9.2 g, 29.3 mmol) was added to 200 mL of xylene at room temperature, followed by PPTS (735 mg, 2.9 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (2E) (6.5 g, yield 74.9%).

[0218] LC-MS, M / Z (ESI): 297.2 [M+H] +

[0219] Step 5: Synthesis of methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (2F)

[0220]

[0221] Methyl 3-(5-bromo-1-methyl-1H-benzo[d]indol-2-yl)propionate (10.0 g, 34 mmol) was added to 100 mL of concentrated sulfuric acid at room temperature. The mixture was cooled to 0 °C, and potassium nitrate (3.6 g, 35 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (2F) (3.0 g, yield 26.0%).

[0222] LC-MS, M / Z (ESI): 342.2 [M+H] +

[0223] Step 6: Synthesis of (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (2G)

[0224]

[0225] At room temperature, methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (2.0 g, 5.8 mmol) was added to 30 mL of dioxane. Under nitrogen protection, Xantphos (1.0 g, 1.7 mmol) and Pd2(dba)3 (1.1 g, 1.2 mmol) were added, along with cesium carbonate (3.8 g, 11.7 mmol) and (R)-pyrrolidine-3-ol (1.0 g, 11.7 mmol). The mixture was heated to 100 °C and stirred for 16 h. Cool to room temperature, add water (100 ml), extract with EA (50 ml × 3), combine organic phases, dry with anhydrous sodium sulfate, concentrate, and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (2 G) (300 mg, yield 14.7%).

[0226] LC-MS, M / Z (ESI): 349.6 [M+H] +

[0227] Step 7: Synthesis of (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (2H)

[0228]

[0229] At room temperature, methyl (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (330 mg, 0.95 mmol) was added to 3 ml THF, 3 ml water and 3 ml ethanol, along with iron powder (213 mg, 3.8 mmol) and ammonium chloride (254 mg, 4.7 mmol). The mixture was heated to 70 °C and stirred for 3 h. Cool to room temperature, add water (10 ml), extract with ethyl acetate (10 ml × 3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate, and separate and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (2H) (200 mg, yield 72.9%).

[0230] LC-MS, M / Z (ESI): 319.6 [M+H] +

[0231] Step 8: Synthesis of (R)-3-(5-(3-hydroxypyrrolidin-1-yl)-1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamide)-1H-benzo[d]imidazol-2-yl)propionate (2I)

[0232]

[0233] Methyl (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (1.25 g, 3.9 mmol) was added to 15 ml DMF and 5 ml DCM at room temperature, along with HATU (2.24 g, 5.9 mmol) and DIPEA (6.5 ml, 39.3 mmol), and 2-(2-methylpyridin-4-yl)oxazol-4-carboxylic acid (963 mg, 4.7 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid (R)-3-(5-(3-hydroxypyrrolidin-1-yl)-1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamide)-1H-benzo[d]imidazol-2-yl)propionate (2I) (1.2 g, yield 60.4%).

[0234] LC-MS, M / Z (ESI): 505.5 [M+H] +

[0235] Step 9: Synthesis of (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (I-2)

[0236]

[0237] At room temperature, methyl (R)-3-(5-(3-hydroxypyrrolidin-1-yl)-1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamide)-1H-benzo[d]indol-2-yl)propionate (1.1 g, 2.2 mmol) was added to 50 mL of dry THF. The mixture was cooled to 0 °C, and a lanthanum(III) chloride bis(lithium chloride) complex solution (0.6 mol / L, 35 mL) and methyl magnesium bromide (1.0 mol / L, 22 mL) were added. The mixture was stirred at room temperature for 16 h. Add 5 ml of water, adjust pH to 6 with 1 mol / L hydrochloric acid, concentrate, and prepare (preparation conditions: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm. Mobile phase A is 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution. Gradient conditions: 0-3 min, mobile phase A maintains 90%, 3-18 min gradient elution, from 90% to 5%, 18-22 min maintains 5%) to give a white solid (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (I-2) (800 mg, yield 72.7%).

[0238] 1H NMR(400MHz,DMSO-d6)δ10.1(s,1H),9.08(s,1H),8.72(d,1H),8.35(s,1H),7.95(s,1H),7.85(d,1H),7.36(s,1H),5.33(t,1H),4 .46(d,2H),3.92(s,3H),3.45(t,2H),3.19(t,3H),3.10(d,1H),2.62(s,3H),2.20(t,1H),2.00(t,1H),1.90(t,2H),1.20(s,6H).

[0239] LC-MS, M / Z (ESI): 505.5 [M+H] +

[0240] Example 3: Preparation of target compound I-3

[0241] N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (target compound I-3)

[0242]

[0243] The synthetic route for the target compound I-3 is shown below:

[0244]

[0245] Step 1: Synthesis of 4-methoxy-N-methyl-2-nitroaniline (3B)

[0246]

[0247] At room temperature, 10.0 g (58.5 mmol) of 4-methoxy-1-fluoro-2-nitrobenzene was added to 50 mL of methylamine methanol solution (containing 7.3 g (233.9 mmol) of methylamine). The mixture was heated to 70 °C and stirred for 32 h. After cooling to room temperature, 100 mL of water was added, and the mixture was extracted with EA (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain a pale yellow crude product of 4-methoxy-N-methyl-2-nitrobenzene (3B) (10.0 g, yield 93.9%).

[0248] LC-MS, M / Z (ESI): 183.4 [M+H] +

[0249] Step 2: 4-Methoxy-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (3C)

[0250]

[0251] At room temperature, 4-methoxy-N-methyl-2-nitroaniline (2 g, 11.0 mmol) was added to 100 mL of THF, followed by 100 mL of ethanol and 50 mL of water. Iron powder (2.5 g, 44.6 mmol) and ammonium chloride (2.5 g, 46.7 mmol) were also added. The mixture was heated to 90 °C and stirred for 2 h. After cooling to room temperature, the mixture was extracted with DCM (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow solid, 4-methoxy-N-methyl-2-nitroaniline. 1 1,2-methylphenyl-1,2-diamine (3C) (1.5 g, yield 89.8%).

[0252] LC-MS, M / Z (ESI): 153.4 [M+H]+ .

[0253] Step 3: Synthesis of methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (3D)

[0254]

[0255] The raw material 4-methoxy-N was prepared at room temperature. 1 1,2-methylphenyl-1,2-diamine (19.5 g, 128 mmol) was added to 1000 ml of DCM and cooled to 0 °C. Methyl 4-chloro-4-oxobutyrate (21 g, 139.5 mmol) and triethylamine (39.0 g, 386.1 mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a colorless crude solid of methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (3D) (18.0 g, yield 52.7%).

[0256] LC-MS, M / Z (ESI): 267.5 [M+H] + .

[0257] Step 4: Synthesis of methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (3E)

[0258]

[0259] Methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (18.0 g, 67.6 mmol) was added to 600 mL of xylene at room temperature, followed by PPTS (1.7 g, 6.8 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (1000 mL) was added, and the mixture was extracted with EA (500 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (3E) (10.5 g, yield 62.5%).

[0260] LC-MS, M / Z (ESI): 249.4 [M+H] + .

[0261] Step 5: Synthesis of methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (3F)

[0262]

[0263] Methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (9.8 g, 39.5 mmol) was added to 50 mL of acetic acid at room temperature, cooled to 0 °C, and 50 mL of nitric acid was added. The mixture was stirred at low temperature for 1 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (3F) (3.8 g, yield 32.8%).

[0264] LC-MS, M / Z (ESI): 294.5 [M+H] + .

[0265] Step 6: Synthesis of 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionic acid (3G)

[0266]

[0267] Methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (1.5 g, 5.1 mmol) was added to 30 mL of pyridine at room temperature. Lithium iodide (10.2 g, 76.1 mmol) was added under nitrogen protection, and the mixture was heated to 150 °C and stirred for 35 h. After cooling to room temperature, the mixture was concentrated, and 1 N hydrochloric acid (100 mL) was added. The mixture was extracted with EA (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (3 G) (1.1 g, yield 80.7%).

[0268] LC-MS, M / Z (ESI): 266.6 [M+H] + .

[0269] Step 7: Synthesis of methyl 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (3H)

[0270]

[0271] 5 ml of thionyl chloride was added to 20 ml of methanol at 0 °C and stirred for 0.5 h. Then, 1.1 g (4.1 mmol) of 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionic acid was added and stirred at room temperature for 3 h. The mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 2:1) to give methyl 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (3H) (900 mg, yield 77.7%).

[0272] LC-MS, M / Z (ESI): 280.6 [M+H] + .

[0273] Step 8: Synthesis of methyl 3-(1-methyl-6-nitro-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (3I)

[0274]

[0275] At room temperature, 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionic acid (900 mg, 3.2 mmol) was added to THF (10 ml), along with triphenylphosphine (1.26 g, 4.8 mmol) and DEAD (840 mg, 4.8 mmol). The mixture was stirred at room temperature for 16 h. The residue was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 2:1) to give methyl 3-(1-methyl-6-nitro-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (3I) (870 mg, yield 77.2%).

[0276] LC-MS, M / Z (ESI): 350.6 [M+H] +

[0277] Step 9: Synthesis of methyl 3-(6-amino-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (3J)

[0278]

[0279] Methyl 3-(1-methyl-6-nitro-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (400 mg, 1.2 mmol) was added to 10 mL of methanol at room temperature, along with 40 mg of palladium on carbon. H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 6 h. The mixture was then filtered and concentrated to give crude white solid methyl 3-(6-amino-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (3J) (300 mg, yield 82.0%).

[0280] LC-MS, M / Z (ESI): 320.6 [M+H] + .

[0281] Step 10: Synthesis of methyl 3-(1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamide)-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (3K)

[0282]

[0283] Methyl 3-(6-amino-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (130 mg, 0.40 mmol) was added to 10 ml of DCM at room temperature, along with HATU (187 mg, 0.49 mmol) and DIPEA (72 mg, 0.56 mmol), and 2-(2-methylpyridin-4-yl)oxazol-4-carboxylic acid (84 mg, 0.41 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamide)-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (3K) (85 mg, yield 41.3%).

[0284] LC-MS, M / Z (ESI): 506.5 [M+H] + .

[0285] Step 11: Synthesis of N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (I-3)

[0286]

[0287] At room temperature, methyl 3-(1-methyl-6-(2-(2-methylpyridin-4-yl)oxazol-4-carboxamide)-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (85 mg, 0.17 mmol) was added to 5 ml of dry THF, cooled to 0 °C, and then lanthanum(III) chloride bis(lithium chloride) complex solution (0.6 mol / L, 1 ml) and methyl magnesium bromide (1.0 mol / L, 1.7 ml) were added. The mixture was stirred at room temperature for 20 h. Add 5 ml of water, adjust pH to 6 with 1 mol / L hydrochloric acid, concentrate, and prepare the separation method as follows (chromatographic column: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm. Mobile phase A: 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution. Gradient conditions: 0-3 min, mobile phase A is maintained at 90%, 3-18 min gradient elution, from 90% to 5%, 18-22 min maintain 5%), to obtain the title compound, white solid N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-6-yl)-2-(2-methylpyridin-4-yl)oxazol-4-carboxamide (I-3) (8.0 mg, yield 9.4%).

[0288] 1 H NMR(400MHz,DMSO-d6)δ9.74(s,1H),9.05(s,1H),8.70(d,1H),8.48(s,1H),7.91(s,1H),7.79(d,1H),7.36(s,1H),4.8 5-4.77(m,4H),4.47(s,1H),4.34(d,2H),3.73(s,3H),3.64(t,1H),2.91(q,2H),2.59(s,3H),1.88(t,2H),1.19(s,6H).

[0289] LC-MS, M / Z (ESI): 506.5 [M+H] + .

[0290] Example 4: Preparation of target compound I-4

[0291] N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholino-1-hydro-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridine-2-carboxamide (target compound I-4)

[0292]

[0293] The synthetic route for the target compound I-4 is as follows:

[0294]

[0295] The synthesis of compound 4A is described in Example 1;

[0296] Step 1: Synthesis of methyl 3-(1-methyl-5-morpholino-6-(6-(trifluoromethyl)pyridin-2-carboxamido)-1-hydro-benzo[d]imidazol-2-yl)propionate (4B)

[0297]

[0298] Methyl 3-(6-amino-1-methyl-5-morpholino-1-hydro-benzo[d]imidazol-2-yl)propionate (240 mg, 0.75 mmol), 6-(trifluoromethyl)pyridine-2-carboxylic acid (158 mg, 0.83 mmol), HATU (428 mg, 1.13 mmol), and triethylamine (228 mg, 2.25 mmol) were dissolved in 2.5 ml. In DMF, the reaction was carried out at room temperature for 6 hours. After the reaction was confirmed to be complete by TLC, DMF was removed by vacuum distillation. The residue was diluted with water (20 mL) and extracted three times with DCM (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by column chromatography to obtain the target compound methyl 3-(1-methyl-5-morpholino-6-(6-(trifluoromethyl)pyridin-2-carboxamido)-1-hydro-benzo[d]imidazol-2-yl)propionate (62 mg, yield 16.8%).

[0299] LC-MS, M / Z (ESI): 492.2 [M+H] +

[0300] The second step is the synthesis of N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholino-1-hydro-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridine-2-carboxamide (I-4).

[0301]

[0302] Methyl 3-(1-methyl-5-morpholino-6-(6-(trifluoromethyl)pyridin-2-carboxamido)-1-hydro-benzo[d]imidazol-2-yl)propionate (62 mg, 0.126 mmol) was dissolved in 1 mL of anhydrous THF under ice bath conditions. Magnesium methyl bromide (3 mol / L solution of 2-methyltetrahydrofuran) (0.4 mL, 1.2 mmol) was slowly added, followed by incubation at room temperature overnight. After TLC detection of complete reaction, saturated ammonium chloride was added. The solution was quenched, diluted with water (20 mL), and extracted three times with DCM (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was separated and purified by silica gel plates (DCM:MeOH = 10:1) to obtain the target compound N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholino-1-hydro-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridine-2-carboxamide (1.5 mg, yield 2.4%).

[0303] 1 H NMR (400MHz, CDCl3) δ11.48(s,1H),8.73(s,1H),8.53(d,J=4.0Hz,1H),8.15(t,J=8.0Hz,1H),7.90(d,J=4.0Hz,1H),7.61 (s,1H),3.99(d,J=4.0Hz,4H),3.78(s,3H),3.04(d,J=8.0Hz,2H),2.92-2.95(m,4H),2.13(t,J=8.0Hz,2H),1.33(s,6H).

[0304] LC-MS, M / Z (ESI): 492.3 [M+H] + .

[0305] Example 5: Preparation of target compound I-5

[0306] (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridinecarboxamide (target compound I-5)

[0307]

[0308] The synthetic route for the target product I-5 is shown below:

[0309]

[0310] Step 1: Synthesis of 4-bromo-N-methyl-2-nitroaniline (5B)

[0311]

[0312] At room temperature, 10.0 g (45.4 mmol) of 4-bromo-1-fluoro-2-nitrobenzene was added to 100 mL of THF, followed by 20 mL of ethanol solution of methylamine (29%). The mixture was heated to 50 °C and stirred for 16 h. After cooling to room temperature, 500 mL of water was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 8:1) to give crude 4-bromo-N-methyl-2-nitrobenzene (5B) (10.5 g, 100% yield) as a pale yellow liquid.

[0313] LC-MS, M / Z (ESI): 231.2 [M+H] +

[0314] Step 2: 4-Bromo-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (5C)

[0315]

[0316] At room temperature, 10.5 g (45.4 mmol) of 4-bromo-N-methyl-2-nitroaniline was added to 100 mL of THF, followed by 100 mL of ethanol and 50 mL of water. Iron powder (10.5 g, 187.5 mmol) and ammonium chloride (10.5 g, 196.2 mmol) were also added. The mixture was heated to 90 °C and stirred for 3 h. After cooling to room temperature, the mixture was extracted with DCM (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow liquid, 4-bromo-N-methyl-2-nitroaniline. 1 1,2-methylphenyl-1,2-diamine (5C) (8.8 g, yield 96.3%).

[0317] LC-MS, M / Z (ESI): 201.2 [M+H] +

[0318] Step 3: Synthesis of methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (5D)

[0319]

[0320] The raw material 4-bromo-N was prepared at room temperature. 11,2-methylphenyl-1,2-diamine (8.8 g, 43.7 mmol) was added to 100 mL of DCM and cooled to 0 °C. Methyl 4-chloro-4-oxobutyrate (7.3 g, 48.5 mmol) and triethylamine (13.2 g, 131 mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (500 mL) was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a colorless crude liquid of methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (5D) (9.2 g, yield 66.7%).

[0321] LC-MS, M / Z (ESI): 315.2 [M+H] +

[0322] Step 4: Synthesis of methyl 3-(5-bromo-1-methyl-1H-benzo[d]indol-2-yl)propionate (5E)

[0323]

[0324] Methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (9.2 g, 29.3 mmol) was added to 200 mL of xylene at room temperature, followed by PPTS (735 mg, 2.9 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (5E) (6.5 g, yield 74.9%).

[0325] LC-MS, M / Z (ESI): 297.2 [M+H] +

[0326] Step 5: Synthesis of methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (5F)

[0327]

[0328] Methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (10.0 g, 34 mmol) was added to 100 mL of concentrated sulfuric acid at room temperature. The mixture was cooled to 0 °C, and potassium nitrate (3.6 g, 35 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (5F) (3.0 g, yield 26.0%).

[0329] LC-MS, M / Z (ESI): 342.2 [M+H] +

[0330] Step 6: Synthesis of (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (5G)

[0331]

[0332] At room temperature, methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (2.0 g, 5.8 mmol) was added to 30 mL of dioxane. Under nitrogen protection, Xantphos (1.0 g, 1.7 mmol) and Pd2(dba)3 (1.1 g, 1.2 mmol) were added, along with cesium carbonate (3.8 g, 11.7 mmol) and (R)-pyrrolidine-3-ol (1.0 g, 11.7 mmol). The mixture was heated to 100 °C and stirred for 16 h. Cool to room temperature, add water (100 ml), extract with EA (50 ml × 3), combine organic phases, dry with anhydrous sodium sulfate, concentrate, and separate and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (5 g) (300 mg, yield 14.7%).

[0333] LC-MS, M / Z (ESI): 349.6 [M+H] +

[0334] Step 7: Synthesis of (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (5H)

[0335]

[0336] At room temperature, methyl (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (330 mg, 0.95 mmol) was added to 3 ml THF, 3 ml water and 3 ml ethanol, along with iron powder (213 mg, 3.8 mmol) and ammonium chloride (254 mg, 4.7 mmol). The mixture was heated to 70 °C and stirred for 3 h. Cool to room temperature, add water (10 ml), extract with ethyl acetate (10 ml × 3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate, and separate and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (5H) (200 mg, yield 72.9%).

[0337] LC-MS, M / Z (ESI): 319.6 [M+H] +

[0338] Step 8: Synthesis of (R)-3-(5-(3-hydroxypyrrolidin-1-yl)-1-methyl-6-(trifluoromethyl)pyridinamide)-1H-benzo[d]imidazol-2-yl)propionate (5I)

[0339]

[0340] Methyl (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (110 mg, 0.35 mmol) was added to 5 mL of DMF at room temperature, along with HATU (200 mg, 0.53 mmol) and DIPEA (135 mg, 1.05 mmol), and 6-(trifluoromethyl)pyridineic acid (66 mg, 0.35 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-(trifluoromethyl)pyridineamide)-1H-benzo[d]imidazol-2-yl)propionate (5I) (100 mg, yield 58.8%).

[0341] LC-MS, M / Z (ESI): 492.2 [M+H] +

[0342] Step 9: Synthesis of (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridineamide (I-5)

[0343]

[0344] Methyl (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-(trifluoromethyl)pyridineamide)-1H-benzo[d]indol-2-yl)propionate (100 mg, 0.20 mmol) was added to 4 ml of dry THF at room temperature, cooled to 0 °C, and lithium chloride (129 mg, 3.0 mmol) and methyl magnesium bromide (1.0 mol / L, 2.1 ml) were added. The mixture was stirred at room temperature for 16 h. Add 5 ml of water, adjust pH to 6 with 1 mol / L hydrochloric acid, concentrate, and prepare (Preparation conditions A: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm. Mobile phase is 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution. Gradient conditions: 0–3 min, mobile phase A is maintained at 90%, 3–18 min gradient elution, from 90% to 5%, 18–22 min maintains 5%) to give a white solid (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridinecarboxamide (I-5) (33 mg, yield 33.0%).

[0345] 1H NMR(400MHz,DMSO-d6)δ11.0(s,1H),8.52(s,1H),8.47(d,1H),8.42(t,1H),8.19(s,1H),7.50(s,1H),4.98(d,1H),4.48(d,2H ),3.72(s,3H),3.48(t,1H),3.29(t,1H),3.08(t,1H),2.91(q,2H),2.80(t,1H),2.22-2.17(m,1H),1.86(t,3H),1.17(s,6H).

[0346] LC-MS, M / Z (ESI): 492.2 [M+H] + .

[0347] Example 6: Preparation of target compound I-6

[0348] N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridineamide (target compound I-6)

[0349]

[0350] The synthetic route for the target compound I-6 is shown below:

[0351]

[0352] Step 1: Synthesis of 4-methoxy-N-methyl-2-nitroaniline (6B)

[0353]

[0354] At room temperature, 10.0 g (58.5 mmol) of 4-methoxy-1-fluoro-2-nitrobenzene was added to 50 mL of methylamine methanol solution (containing 7.3 g (233.9 mmol) of methylamine). The solution was heated to 70 °C and stirred for 32 h. After cooling to room temperature, 100 mL of water was added, and the mixture was extracted with EA (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give crude 4-methoxy-N-methyl-2-nitrobenzene (6B) (10.0 g, yield 93.9%) as a pale yellow solid.

[0355] LC-MS, M / Z (ESI): 183.4 [M+H] + .

[0356] Step 2: 4-Methoxy-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (6C)

[0357]

[0358] At room temperature, 4-methoxy-N-methyl-2-nitroaniline (2 g, 11.0 mmol) was added to 100 mL of THF, followed by 100 mL of ethanol and 50 mL of water. Iron powder (2.5 g, 44.6 mmol) and ammonium chloride (2.5 g, 46.7 mmol) were also added. The mixture was heated to 90 °C and stirred for 2 h. After cooling to room temperature, the mixture was extracted with DCM (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow solid, 4-methoxy-N-methyl-2-nitroaniline. 1 -Methylphenyl-1,2-diamine (6C) (1.5g, yield 89.8%).

[0359] LC-MS, M / Z (ESI): 153.4 [M+H] +.

[0360] Step 3: Synthesis of methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (6D)

[0361]

[0362] The raw material 4-methoxy-N was prepared at room temperature. 1 1,2-methylphenyl-1,2-diamine (19.5 g, 128 mmol) was added to 1000 ml of DCM and cooled to 0 °C. Methyl 4-chloro-4-oxobutyrate (21 g, 139.5 mmol) and triethylamine (39.0 g, 386.1 mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a colorless crude solid, methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (6D) (18.0 g, yield 52.7%).

[0363] LC-MS, M / Z (ESI): 267.5 [M+H] + .

[0364] Step 4: Synthesis of methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (6E)

[0365]

[0366] Methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (18.0 g, 67.6 mmol) was added to 600 mL of xylene at room temperature, followed by PPTS (1.7 g, 6.8 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (1000 mL) was added, and the mixture was extracted with EA (500 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (6E) (10.5 g, yield 62.5%) as a white solid.

[0367] LC-MS, M / Z (ESI): 249.4 [M+H] + .

[0368] Step 5: Synthesis of methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (6F)

[0369]

[0370] Methyl 3-(5-methoxy-1-methyl-1H-benzo[d]indol-2-yl)propionate (9.8 g, 39.5 mmol) was added to 50 mL of acetic acid at room temperature, cooled to 0 °C, and 50 mL of nitric acid was added. The mixture was stirred at low temperature for 1 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (6F) (3.8 g, yield 32.8%).

[0371] LC-MS, M / Z (ESI): 294.5 [M+H] + .

[0372] Step 6: Synthesis of 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionic acid (6G)

[0373]

[0374] Methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (1.5 g, 5.1 mmol) was added to 30 mL of pyridine at room temperature. Under nitrogen protection, lithium iodide (10.2 g, 76.1 mmol) was added, and the mixture was heated to 150 °C and stirred for 35 h. After cooling to room temperature, the mixture was concentrated, and 1 N hydrochloric acid (100 mL) was added. The mixture was extracted with EA (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (6 g) (1.1 g, yield 80.7%).

[0375] LC-MS, M / Z (ESI): 266.6 [M+H] + .

[0376] Step 7: Synthesis of methyl 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (6H)

[0377]

[0378] 5 ml of thionyl chloride was added to 20 ml of methanol at 0 °C and stirred for 0.5 h. Then, 1.1 g (4.1 mmol) of 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionic acid was added and stirred at room temperature for 3 h. The mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 2:1) to give methyl 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (6H) (900 mg, yield 77.7%).

[0379] LC-MS, M / Z (ESI): 280.6 [M+H] + .

[0380] Step 8: Synthesis of methyl 3-(1-methyl-6-nitro-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (6I)

[0381]

[0382] At room temperature, 3-(5-hydroxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionic acid (900 mg, 3.2 mmol) was added to THF (10 ml), along with triphenylphosphine (1.26 g, 4.8 mmol) and DEAD (840 mg, 4.8 mmol). The mixture was stirred at room temperature for 16 h. The residue was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give methyl 3-(1-methyl-6-nitro-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (6I) (870 mg, yield 77.2%).

[0383] LC-MS, M / Z (ESI): 350.6 [M+H] +

[0384] Step 9: Synthesis of methyl 3-(6-amino-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (6J)

[0385]

[0386] Methyl 3-(1-methyl-6-nitro-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (400 mg, 1.2 mmol) was added to 10 mL of methanol at room temperature, along with palladium on carbon (40 mg, 10%). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 6 h. After filtration and concentration, the crude product was a white solid, methyl 3-(6-amino-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (6 J) (300 mg, yield 82.0%).

[0387] LC-MS, M / Z (ESI): 320.6 [M+H] +

[0388] Step 10: Synthesis of methyl 3-(1-methyl-5-(epoxypropane-3-ylmethoxy)-6-(6-(trifluoromethyl)pyridinamide)-1H-benzo[d]imidazol-2-yl)propionate (6K)

[0389]

[0390] Methyl 3-(6-amino-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-2-yl)propionate (130 mg, 0.40 mmol) was added to 10 mL of DCM at room temperature, along with HATU (169 mg, 0.44 mmol) and DIPEA (72 mg, 0.56 mmol), and 6-(1-methyl-1H-pyrrole-3-yl)pyridine acid (78 mg, 0.41 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(1-methyl-5-(epoxypropane-3-ylmethoxy)-6-(6-(trifluoromethyl)pyridineamide)-1H-benzo[d]imidazol-2-yl)propionate (6K) (120 mg, yield 59.8%).

[0391] LC-MS, M / Z (ESI): 493.4 [M+H] +

[0392] Step 11: Synthesis of N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridineamide (I-6)

[0393]

[0394] At room temperature, methyl 3-(1-methyl-5-(epoxypropane-3-ylmethoxy)-6-(6-(trifluoromethyl)pyridinamide)-1H-benzo[d]imidazol-2-yl)propionate (120 mg, 0.24 mmol) was added to 5 ml of dry THF, cooled to 0 °C, and then lanthanum(III) chloride bis(lithium chloride) complex solution (0.6 mol / L, 4.0 ml) and methyl magnesium bromide (1.0 mol / L, 2.4 ml) were added. The mixture was stirred at room temperature for 24 h. Add 5 ml of water, adjust pH to 6 with 1 mol / L hydrochloric acid, concentrate, and prepare the separation method as follows (chromatographic column: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm; mobile phase A: 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution; gradient conditions: 0-3 minutes, mobile phase A is maintained at 90%, 3-18 minutes gradient elution, from 90% to 5%, 18-22 minutes maintain 5%), to obtain white solid N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-(epoxypropane-3-ylmethoxy)-1H-benzo[d]imidazol-6-yl)-6-(trifluoromethyl)pyridineamide (I-6) (28.5 mg, yield 23.7%).

[0395] 1H NMR (400MHz, DMSO-d6) δ10.4(s,1H),8.57(s,1H),8.48(t,1H),8.41(d,1H),8.23(d,1H),7.36(s,1H),4.79(q ,2H),4.52(t,2H),4.46(s,1H),4.42(d,2H),3.73(s,3H),3.50(t,1H),2.92(q,2H),1.87(q,2H),1.18(s,6H).

[0396] LC-MS, M / Z (ESI): 493.4 [M+H] + .

[0397] Example 7: Preparation of target compound I-7

[0398] (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-6-(1-methyl-1H-pyrazole-3-yl)pyridinecarboxamide (target compound I-7)

[0399]

[0400] The synthetic route for the target product I-7 is shown below:

[0401]

[0402] Step 1: Synthesis of 4-bromo-N-methyl-2-nitroaniline (7B)

[0403]

[0404] At room temperature, 10.0 g (45.4 mmol) of 4-bromo-1-fluoro-2-nitrobenzene was added to 100 mL of THF, followed by 20 mL of 29% methylamine in ethanol. The mixture was heated to 50 °C and stirred for 16 h. After cooling to room temperature, 500 mL of water was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 8:1) to give crude 4-bromo-N-methyl-2-nitrobenzene (7B) (10.5 g, 100% yield) as a pale yellow liquid.

[0405] LC-MS, M / Z (ESI): 231.2 [M+H] +

[0406] Step 2: 4-Bromo-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (7C)

[0407]

[0408] At room temperature, 10.5 g (45.4 mmol) of 4-bromo-N-methyl-2-nitroaniline was added to 100 mL of THF, followed by 100 mL of ethanol and 50 mL of water. Iron powder (10.5 g, 187.5 mmol) and ammonium chloride (10.5 g, 196.2 mmol) were also added. The mixture was heated to 90 °C and stirred for 3 h. After cooling to room temperature, the mixture was extracted with DCM (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow liquid, 4-bromo-N-methyl-2-nitroaniline. 1 -Methylphenyl-1,2-diamine (7C) (8.8 g, yield 96.3%).

[0409] LC-MS, M / Z (ESI): 201.2 [M+H] +

[0410] Step 3: Synthesis of methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (7D)

[0411]

[0412] The raw material 4-bromo-N was prepared at room temperature. 11,2-methylphenyl-1,2-diamine (8.8 g, 43.7 mmol) was added to 100 mL of DCM and cooled to 0 °C. Methyl 4-chloro-4-oxobutyrate (7.3 g, 48.5 mmol) and triethylamine (13.2 g, 131 mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (500 mL) was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a colorless crude liquid of methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (7D) (9.2 g, yield 66.7%).

[0413] LC-MS, M / Z (ESI): 315.2 [M+H] +

[0414] Step 4: Synthesis of methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (7E)

[0415]

[0416] Methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (9.2 g, 29.3 mmol) was added to 200 mL of xylene at room temperature, followed by PPTS (735 mg, 2.9 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (7E) (6.5 g, yield 74.9%).

[0417] LC-MS, M / Z (ESI): 297.2 [M+H] +

[0418] Step 5: Synthesis of methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (7F)

[0419]

[0420] At room temperature, methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (10.0 g, 34 mmol) was added to 100 mL of concentrated sulfuric acid. The mixture was cooled to 0 °C, and potassium nitrate (3.6 g, 35 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]indol-2-yl)propionate (7F) (3.0 g, yield 26.0%).

[0421] LC-MS, M / Z (ESI): 342.2 [M+H] +

[0422] Step 6: Synthesis of (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (7G)

[0423]

[0424] At room temperature, methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (2.0 g, 5.8 mmol) was added to 30 mL of dioxane. Under nitrogen protection, Xantphos (1.0 g, 1.7 mmol) and Pd2(dba)3 (1.1 g, 1.2 mmol) were added, along with cesium carbonate (3.8 g, 11.7 mmol) and (R)-pyrrolidine-3-ol (1.0 g, 11.7 mmol). The mixture was heated to 100 °C and stirred for 16 h. Cool to room temperature, add water (100 ml), extract with EA (50 ml × 3), combine organic phases, dry with anhydrous sodium sulfate, concentrate, and separate and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (7 g) (300 mg, yield 14.7%).

[0425] LC-MS, M / Z (ESI): 349.6 [M+H] +

[0426] Step 7: Synthesis of (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]indol-2-yl)propionate (7H)

[0427]

[0428] At room temperature, methyl (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (330 mg, 0.95 mmol) was added to 3 ml THF, 3 ml water and 3 ml ethanol, along with iron powder (213 mg, 3.8 mmol) and ammonium chloride (254 mg, 4.7 mmol). The mixture was heated to 70 °C and stirred for 3 h. Cool to room temperature, add water (10 ml), extract with ethyl acetate (10 ml × 3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate, and separate and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give a white solid (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (7H) (200 mg, yield 72.9%).

[0429] LC-MS, M / Z (ESI): 319.6 [M+H] +

[0430] Step 8: Synthesis of (R)-3-(5-(3-hydroxypyrrolidin-1-yl)-1-methyl-6-(6-(1-methyl-1H-pyrazole-3-yl)pyridinamide)-1H-benzo[d]imidazol-2-yl)propionate (7I)

[0431]

[0432] Methyl (R)-3-(6-amino-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (100 mg, 0.31 mmol) was added to 5 ml of DMF at room temperature, along with HATU (180 mg, 0.47 mmol) and DIPEA (122 mg, 0.95 mmol), and 6-(1-methyl-1H-pyrrole-3-yl)pyridine acid (64 mg, 0.31 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-(6-(1-methyl-1H-pyrazole-3-yl)pyridineamide)-1H-benzo[d]imidazol-2-yl)propionate (7I) (65 mg, yield 41.0%).

[0433] LC-MS, M / Z (ESI): 504.4 [M+H] +

[0434] Step 9: Synthesis of (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-6-(1-methyl-1H-pyrazole-3-yl)pyridineamide (I-7)

[0435]

[0436] Methyl (R)-3-(5-(3-hydroxypyrrolidone-1-yl)-1-methyl-6-(6-(1-methyl-1H-pyrazol-3-yl)pyridineamide)-1H-benzo[d]indol-2-yl)propionate (65 mg, 0.13 mmol) was added to 5 ml of dry THF at room temperature, cooled to 0 °C, and lithium chloride (110 mg, 2.6 mmol) and methyl magnesium bromide (1.0 mol / L, 2.6 ml) were added. The mixture was stirred at room temperature for 48 h. Add 5 ml of water, adjust pH to 6 with 1 mol / L hydrochloric acid, concentrate, and prepare (Preparation conditions A: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm. Mobile phase A is 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution. Gradient conditions: 0–3 min, mobile phase A maintains 90%, 3–18 min gradient elution, from 90% to 5%, 18–22 min maintains 5%) to give a white solid (R)-N-(2-(3-hydroxy-3-methylbutyl)-5-(3-hydroxypyrrolidone-1-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-6-(1-methyl-1H-pyrazole-3-yl)pyridinecarboxamide (I-7) (15 mg, yield 23.0%).

[0437] 1H NMR(400MHz,DMSO-d6)δ11.1(s,1H),8.57(s,1H),8.13(t,3H),7.87(d,1H),7.50(s,1H),7.07(d,1H),5.02(d,1H),4.50(d,1H),4.4 7(s,1H),3.97(s,3H),3.72(s,3H),3.49(t,1H),3.25(t,1H),3.11(t,1H),2.91(q,3H),2.26-2.24(m,1H),1.87(t,3H),1.18(s,6H).

[0438] LC-MS, M / Z (ESI): 504.4 [M+H] +

[0439] Example 8: Preparation of target compound I-8

[0440] N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholin-1H-benzo[d]imidazol-6-yl)-6-(1-methyl-1H-pyrazol-3-yl)pyridinecarboxamide (target compound I-8)

[0441]

[0442] The synthetic route for the target compound I-8 is shown below:

[0443]

[0444] Step 1: Synthesis of 4-bromo-N-methyl-2-nitroaniline (8B)

[0445]

[0446] At room temperature, 10.0 g (45.4 mmol) of 4-bromo-1-fluoro-2-nitrobenzene was added to 100 mL of THF, followed by 20 mL of 29% methylamine in ethanol. The mixture was heated to 50 °C and stirred for 16 h. After cooling to room temperature, 500 mL of water was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 8:1) to give crude 4-bromo-N-methyl-2-nitrobenzene (8B) (10.5 g, 100% yield) as a pale yellow liquid.

[0447] LC-MS, M / Z (ESI): 231.2 [M+H] + .

[0448] Step 2: 4-Bromo-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (8C)

[0449]

[0450] At room temperature, 10.5 g (45.4 mmol) of 4-bromo-N-methyl-2-nitroaniline was added to 100 mL of THF, followed by 100 mL of ethanol and 50 mL of water. Iron powder (10.5 g, 187.5 mmol) and ammonium chloride (10.5 g, 196.2 mmol) were also added. The mixture was heated to 90 °C and stirred for 3 h. After cooling to room temperature, the mixture was extracted with DCM (150 mL × 3). The organic phases were combined, concentrated, dried over anhydrous sodium sulfate, and concentrated again. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow liquid, 4-bromo-N-methyl-2-nitroaniline. 1 1,2-methylphenyl-1,2-diamine (8C) (8.8 g, yield 96.3%).

[0451] LC-MS, M / Z (ESI): 201.2 [M+H] +

[0452] Step 3: Synthesis of methyl 4-((2-amino-4-bromophenyl)(methyl)amino)-4-oxobutyrate (8D)

[0453]

[0454] The raw material 4-bromo-N was prepared at room temperature. 1 1,2-methylphenyl-1,2-diamine (8.8 g, 43.7 mmol) was added to 100 mL of DCM and cooled to 0 °C. Methyl 4-chloro-4-oxobutyrate (7.3 g, 48.5 mmol) and triethylamine (13.2 g, 131 mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (500 mL) was added, and the mixture was extracted with DCM (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a colorless crude liquid of methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (8D) (9.2 g, yield 66.7%).

[0455] LC-MS, M / Z (ESI): 315.2 [M+H] +

[0456] Step 4: Synthesis of methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (8E)

[0457]

[0458] Methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (9.2 g, 29.3 mmol) was added to 200 mL of xylene at room temperature, followed by PPTS (735 mg, 2.9 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (8E) (6.5 g, yield 74.9%).

[0459] LC-MS, M / Z (ESI): 297.2 [M+H] +

[0460] Step 5: Synthesis of methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (8F)

[0461]

[0462] Methyl 3-(5-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (10.0 g, 34 mmol) was added to 100 mL of concentrated sulfuric acid at room temperature. The mixture was cooled to 0 °C, and potassium nitrate (3.6 g, 35 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (8F) (3.0 g, yield 26.0%).

[0463] LC-MS, M / Z (ESI): 342.2 [M+H] +

[0464] Step 6: Synthesis of methyl 3-(1-methyl-5-morpholino-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (8G)

[0465]

[0466] At room temperature, methyl 3-(5-bromo-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (4.0 g, 11.7 mmol) was added to 120 ml of dioxane. Under nitrogen protection, Ruphos-G3 (963 mg, 1.17 mmol) and Pd2(dba)3 (667 mg, 1.17 mmol) were added, along with cesium carbonate (11.2 g, 34.4 mmol) and morpholine (2.0 g, 23.0 mmol). The mixture was heated to 100 °C and stirred for 16 h under nitrogen protection. Cool to room temperature, add water (400 ml), extract with EA (500 ml × 3), combine organic phases, dry with anhydrous sodium sulfate, concentrate, and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give methyl 3-(1-methyl-5-morpholino-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (8 g) (3.15 g, yield 77.1%).

[0467] LC-MS, M / Z (ESI): 349.6 [M+H] +

[0468] Step 7: Synthesis of methyl 3-(6-amino-1-methyl-5-morpholino-1H-benzo[d]imidazol-2-yl)propionate (8H)

[0469]

[0470] Methyl 3-(1-methyl-5-morpholino-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (350 mg, 1.0 mmol) was added to 10 mL of methanol at room temperature, along with palladium on carbon (35 mg). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was then filtered and concentrated to give crude white solid methyl 3-(6-amino-1-methyl-5-morpholino-1H-benzo[d]imidazol-2-yl)propionate (8H) (270 mg, yield 84.4%).

[0471] LC-MS, M / Z (ESI): 319.6 [M+H] +

[0472] Step 8: Synthesis of methyl 3-(1-methyl-6-(6-(1-methyl-1H-pyrazole-3-yl)pyridinecarboxamide)-5-morpholine-1H-benzo[d]imidazol-2-yl)propionate (8I)

[0473]

[0474] Methyl 3-(6-amino-1-methyl-5-morpholino-1H-benzo[d]imidazol-2-yl)propionate (140 mg, 0.44 mmol) was added to 5 mL of DMF at room temperature, followed by HATU (250 mg, 0.66 mmol) and DIPEA (130 mg, 0.90 mmol), and then 6-(1-methyl-1H-pyrazole-3-yl)pyridinecarboxylic acid (90 mg, 0.45 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(1-methyl-6-(6-(1-methyl-1H-pyrazole-3-yl)pyridinecarboxamide)-5-morpholino-1H-benzo[d]imidazol-2-yl)propionate (8I) (130 mg, yield 58.5%).

[0475] LC-MS, M / Z (ESI): 504.3 [M+H] +

[0476] Step 9: Synthesis of N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholin-1H-benzo[d]imidazol-6-yl)-6-(1-methyl-1H-pyrazol-3-yl)pyridinecarboxamide (I-8)

[0477]

[0478] Methyl 3-(1-methyl-6-(6-(1-methyl-1H-pyrazol-3-yl)pyridineamide)-5-morpholino-1H-benzo[d]imidazol-2-yl)propionate (130 mg, 0.26 mmol) was added to 2 ml of dry THF at room temperature, cooled to 0 °C, and methyl magnesium bromide (1.0 mol / L, 3 ml) was added. The mixture was stirred at room temperature for 16 h. Add 5 ml of water, adjust pH to 6 with 1 mol / L hydrochloric acid, concentrate, and prepare the separation method as follows (chromatographic column: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm. Mobile phase A: 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution. Gradient conditions: 0-3 min, mobile phase A is maintained at 90%, 3-18 min gradient elution, from 90% to 5%, 18-22 min maintain 5%), to obtain white solid N-(2-(3-hydroxy-3-methylbutyl)-1-methyl-5-morpholin-1H-benzo[d]imidazol-6-yl)-6-(1-methyl-1H-pyrazol-3-yl)pyridineamide (I-8) (25 mg, yield 19.2%).

[0479] 1 H NMR(400MHz,DMSO-d6)δ11.2(s,1H),8.62(s,1H),8.13(d,3H),7.93(d,1H),7.52(s,1H),7.10(d,1H ),4.47(s,1H),3.99(s,3H),3.92(d,4H),3.74(s,3H),2.81-2.72(m,6H),1.88(t,2H),1.18(s,6H).

[0480] LC-MS, M / Z (ESI): 504.3 [M+H] + .

[0481] Example 9: Preparation of target compound I-9

[0482] 2-Fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-3-(trifluoromethyl)benzamide (Target compound I-9)

[0483]

[0484] The synthetic route for the target product I-9 is shown below:

[0485]

[0486] Step 1: Synthesis of methyl 4-(methylamino)-3-nitrobenzene (9B)

[0487]

[0488] At room temperature, methyl 4-fluoro-3-nitrobenzene (40.0 g, 200 mmol) was added to 600 mL of THF, followed by an ethanol solution of methylamine (29%, 150 mL). The mixture was heated to 70 °C and stirred for 72 h. After cooling to room temperature, water (1500 mL) was added, and the mixture was extracted with DCM (1200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give crude methyl 4-(methylamino)-3-nitrobenzene (9B) (30.0 g, yield 71.0%) as a pale yellow solid.

[0489] LC-MS, M / Z (ESI): 211.2 [M+H] +

[0490] Step 2: Synthesis of methyl 3-amino-4-(methylamino)benzoate (9C)

[0491]

[0492] At room temperature, methyl 4-(methylamino)-3-nitrobenzene (25.0 g, 119 mmol) was added to 300 mL of methanol, followed by the addition of 10% Pd / C (2.5 g). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow solid methyl 3-amino-4-(methylamino)benzoate (9C) (18.0 g, yield 84.0%).

[0493] LC-MS, M / Z (ESI): 181.2 [M+H] +

[0494] Step 3: Synthesis of methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (9D)

[0495]

[0496] At room temperature, methyl 3-amino-4-(methylamino)benzoate (18.0 g, 100 mmol) was added to 500 ml of DCM. The mixture was cooled to 0 °C, and methyl 4-chloro-4-oxobutyrate (18.0 g, 120 mmol) and triethylamine (30.0 g, 300 mmol) were added. The mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give a colorless crude solid methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (9D) (13.0 g, yield 66.7%).

[0497] LC-MS, M / Z (ESI): 295.6 [M+H] +

[0498] Step 4: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (9E)

[0499]

[0500] Methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (9D) (7.0 g, 23.8 mmol) was added to 300 mL of xylene at room temperature, followed by the addition of PPTS (1.8 g, 7.1 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (300 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (9E) (4.0 g, yield 60.8%).

[0501] LC-MS, M / Z (ESI): 277.4 [M+H] +

[0502] Step 5: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (9F)

[0503]

[0504] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (4.0 g, 14.5 mmol) was added to 100 mL of concentrated sulfuric acid at room temperature, cooled to 0 °C, and potassium nitrate (2.9 g, 29 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (9F) (1.4 g, yield 30.1%).

[0505] LC-MS, M / Z (ESI): 322.4 [M+H] +

[0506] Step 6: Synthesis of methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (9G)

[0507]

[0508] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (2.0 g, 6.2 mmol) was added to 50 mL of methanol at room temperature, followed by the addition of 10% Pd / C (200 mg). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:1) to give crude methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (9 g) (1.8 g, yield 99.2%) as a white solid.

[0509] LC-MS, M / Z (ESI): 292.6 [M+H] +

[0510] Step 7: Synthesis of methyl 6-(2-fluoro-3-(trifluoromethyl)benzamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (9H)

[0511]

[0512] 1.8 g (6.2 mmol) of methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate was added to 50 mL of DCM at room temperature, followed by triethylamine (1.9 g, 18.6 mmol) and 2-fluoro-3-(trifluoromethyl)benzoyl chloride (2.1 g, 10.0 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:1) to give a white solid methyl 6-(2-fluoro-3-(trifluoromethyl)benzoamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (9H) (2.1 g, yield 70.5%).

[0513] LC-MS, M / Z (ESI): 482.6 [M+H] +

[0514] Step 8: Synthesis of 2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-3-(trifluoromethyl)benzamide (I-9)

[0515]

[0516] At room temperature, methyl 6-(2-fluoro-3-(trifluoromethyl)benzamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]indole-5-carboxylate (2.0 g, 4.1 mmol) was added to 40 mL of dry THF. The mixture was cooled to 0 °C, and then lanthanum(III) chloride bis(lithium chloride) complex solution (0.6 mol / L, 44 mL) and methyl magnesium bromide (1.0 mol / L, 44 mL) were added. The mixture was stirred at room temperature for 16 h. Add 50 ml of saturated ammonium chloride solution, extract with EA (80 ml × 3), dry, filter and concentrate, and prepare (preparation conditions: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm. Mobile phase A is 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution. Gradient conditions: 0-3 min, mobile phase A is maintained at 90%, 3-18 min gradient elution, from 90% to 5%, 18-22 min maintain 5%) to give a white solid 2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-3-(trifluoromethyl)benzamide (I-9) (780 mg, yield 33.0%).

[0517] 1H NMR(400MHz,DMSO-d6)δ11.6(s,1H),8.39(s,1H),8.19(t,1H),8.01(t,1H),7.60(t,1H),7.50(s ,1H),6.18(s,1H),4.46(d,1H),3.72(s,3H),2.92(t,2H),1.87(t,2H),1.59(t,6H),1.17(s,6H).

[0518] LC-MS, M / Z (ESI): 482.5 [M+H] + .

[0519] Example 10: Preparation of target compound I-10

[0520] N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-methyl-3-(trifluoromethyl)benzamide (target compound I-10)

[0521]

[0522] The synthetic route for the target compound I-10 is shown below:

[0523]

[0524] Step 1: Synthesis of methyl 4-(methylamino)-3-nitrobenzene (10B)

[0525]

[0526] At room temperature, methyl 4-fluoro-3-nitrobenzene (40.0 g, 200 mmol) was added to 600 mL of THF, followed by an ethanol solution of methylamine (29%, 150 mL). The mixture was heated to 70 °C and stirred for 72 h. After cooling to room temperature, water (1500 mL) was added, and the mixture was extracted with DCM (1200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give the title compound, a pale yellow crude solid, methyl 4-(methylamino)-3-nitrobenzene (30.0 g, yield 71.0%).

[0527] LC-MS, M / Z (ESI): 211.2 [M+H] +

[0528] Step 2: Synthesis of methyl 3-amino-4-(methylamino)benzoate (10C)

[0529]

[0530] At room temperature, methyl 4-(methylamino)-3-nitrobenzene (25.0 g, 119 mmol) was added to 300 mL of methanol, followed by the addition of 10% Pd / C (2.5 g). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give the title compound, a pale yellow solid, methyl 3-amino-4-(methylamino)benzoate (18.0 g, yield 84.0%).

[0531] LC-MS, M / Z (ESI): 181.2 [M+H] +

[0532] Step 3: Synthesis of methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (10D)

[0533]

[0534] At room temperature, methyl 3-amino-4-(methylamino)benzoate (18.0 g, 100 mmol) was added to 500 ml of DCM. The mixture was cooled to 0 °C, and methyl 4-chloro-4-oxobutyrate (18.0 g, 120 mmol) and triethylamine (30.0 g, 300 mmol) were added. The mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give the title compound, a colorless crude solid, methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide) benzoate (13.0 g, yield 66.7%).

[0535] LC-MS, M / Z (ESI): 295.6 [M+H] +

[0536] Step 4: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (10E)

[0537]

[0538] Methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (7.0 g, 23.8 mmol) was added to 300 mL of xylene at room temperature, along with pyridinium p-toluenesulfonate (1.8 g, 7.1 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (300 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give the title compound, a white solid, methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]indole-5-carboxylate (4.0 g, yield 60.8%).

[0539] LC-MS, M / Z (ESI): 277.4 [M+H] +

[0540] Step 5: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (10F)

[0541]

[0542] 4.0 g (14.5 mmol) of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate was added to 100 mL of concentrated sulfuric acid at room temperature. The mixture was cooled to 0 °C, and potassium nitrate (2.9 g, 29 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give the title compound, a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (1.4 g, yield 30.1%).

[0543] LC-MS, M / Z (ESI): 322.4 [M+H] +

[0544] Step 6: Synthesis of methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (10G)

[0545]

[0546] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (2.0 g, 6.2 mmol) was added to 50 mL of methanol at room temperature, followed by the addition of 10% Pd / C (200 mg). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:1) to give the title compound, a white solid crude methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (1.8 g, 99.2% yield).

[0547] LC-MS, M / Z (ESI): 292.6 [M+H] +

[0548] Step 7: Synthesis of 2-fluoro-3-(trifluoromethyl)benzoyl chloride (10H)

[0549]

[0550] At room temperature, 2-fluoro-3-(trifluoromethyl)benzoic acid (2.0 g, 9.6 mmol) was added to 50 mL of dry THF, followed by 3 drops of DMF. The mixture was cooled to 0 °C, and then oxaloyl chloride (1.9 g, 15 mmol) was added. The mixture was stirred at room temperature for 2 h. The reaction solution was concentrated to obtain the title compound, a white solid crude 2-methyl-3-(trifluoromethyl)benzoyl chloride (2.1 g), which was used directly in the next reaction step.

[0551] Step 8: Synthesis of methyl 6-(2-fluoro-3-(trifluoromethyl)benzamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (10I)

[0552]

[0553] 1.8 g (6.2 mmol) of methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate was added to 50 mL of DCM at room temperature, followed by triethylamine (1.9 g, 18.6 mmol) and 2-fluoro-3-(trifluoromethyl)benzoyl chloride (2.1 g, 10.0 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:1) to give the title compound, a white solid, methyl 6-(2-fluoro-3-(trifluoromethyl)benzoamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (2.1 g, yield 70.5%).

[0554] LC-MS, M / Z (ESI): 482.6 [M+H] +

[0555] Step 9: Synthesis of N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-methyl-3-(trifluoromethyl)benzamide (I-10)

[0556]

[0557] At room temperature, methyl 6-(2-fluoro-3-(trifluoromethyl)benzamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (2.0 g, 1.0 mmol) was added to 40 mL of dry THF. The mixture was cooled to 0 °C, and lithium chloride and lanthanum chloride solution (0.6 mol / L, 72 mL) and methyl magnesium bromide (1.0 mol / L, 44 mL) were added. The mixture was stirred at room temperature for 16 h. Add 100 ml of saturated ammonium chloride solution, extract with EA (80 ml × 3), dry, filter and concentrate, and purify by acid preparation method A to obtain the title compound, white solid N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-methyl-3-(trifluoromethyl)benzamide (I-10) (15 mg, yield 0.75%).

[0558] 1 H NMR(400MHz,DMSO-d6)δ11.24(s,1H),8.45(s,1H),7.84(d,1H),7.79(d,1H),7.56(t,1H),7.49(s,1H), 6.14(s,1H),4.45(s,1H),3.72(s,3H),2.92(t,2H),2.53(d,3H),1.87(t,2H),1.58(s,6H),1.17(s,6H).

[0559] LC-MS, M / Z (ESI): 478.5 [M+H] + .

[0560] Example 11: Preparation of target compound I-11

[0561] 2-Fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-3-methylbenzamide (Target compound I-11)

[0562]

[0563] The synthetic route for the target compound I-11 is shown below:

[0564]

[0565] Step 1: Synthesis of methyl 4-(methylamino)-3-nitrobenzene (11B)

[0566]

[0567] At room temperature, methyl 4-fluoro-3-nitrobenzene (40.0 g, 200 mmol) was added to 600 mL of THF, followed by an ethanol solution of methylamine (29%, 150 mL). The mixture was heated to 70 °C and stirred for 72 h. After cooling to room temperature, water (1500 mL) was added, and the mixture was extracted with DCM (1200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give crude methyl 4-(methylamino)-3-nitrobenzene (11B) (30.0 g, yield 71.0%) as a pale yellow solid.

[0568] LC-MS, M / Z (ESI): 211.2 [M+H] +

[0569] Step 2: Synthesis of methyl 3-amino-4-(methylamino)benzoate (11C)

[0570]

[0571] At room temperature, methyl 4-(methylamino)-3-nitrobenzene (25.0 g, 119 mmol) was added to 300 mL of methanol, followed by the addition of 10% Pd / C (2.5 g). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow solid methyl 3-amino-4-(methylamino)benzoate (11C) (18.0 g, yield 84.0%).

[0572] LC-MS, M / Z (ESI): 181.2 [M+H] + .

[0573] Step 3: Synthesis of methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (11D)

[0574]

[0575] At room temperature, methyl 3-amino-4-(methylamino)benzoate (18.0 g, 100 mmol) was added to 500 ml of DCM. The mixture was cooled to 0 °C, and methyl 4-chloro-4-oxobutyrate (18.0 g, 120 mmol) and triethylamine (30.0 g, 300 mmol) were added. The mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give a colorless crude solid methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (11D) (13.0 g, yield 66.7%).

[0576] LC-MS, M / Z (ESI): 295.6 [M+H] + .

[0577] Step 4: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (11E)

[0578]

[0579] Methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (7.0 g, 23.8 mmol) was added to 300 mL of xylene at room temperature, followed by PPTS (1.8 g, 7.1 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (300 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (11E) (4.0 g, yield 60.8%).

[0580] LC-MS, M / Z (ESI): 277.4 [M+H] +

[0581] Step 5: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (11F)

[0582]

[0583] 4.0 g (14.5 mmol) of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate was added to 100 mL of concentrated sulfuric acid at room temperature. The mixture was cooled to 0 °C, and potassium nitrate (2.9 g, 29 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (11F) (1.4 g, yield 30.1%).

[0584] LC-MS, M / Z (ESI): 322.4 [M+H] +

[0585] Step 6: Synthesis of methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (11G)

[0586]

[0587] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (2.0 g, 6.2 mmol) was added to 50 mL of methanol at room temperature, along with 10% Pd / C (200 mg). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:1) to give crude methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (11 g) (1.8 g, yield 99.2%).

[0588] LC-MS, M / Z (ESI): 292.6 [M+H] + .

[0589] Step 7: Synthesis of 4-(6-amino-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (11H)

[0590]

[0591] At room temperature, methyl 6-(2-fluoro-3-methylbenzamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazol-5-carboxylate (300 mg, 1.0 mmol) was added to 3 mL of dry THF. The mixture was cooled to 0 °C, and a lanthanum(III) chloride bis(lithium chloride) complex solution (0.6 mol / L, 1.7 mL) and methyl magnesium bromide (1.0 mol / L, 10.3 mL) were added. The mixture was stirred at room temperature for 16 h. 50 mL of saturated ammonium chloride solution was added, and the mixture was extracted with EA (80 mL × 3). The extract was dried, filtered, concentrated, and purified by thin-layer chromatography to obtain a white solid 4-(6-amino-5-(2-hydroxypropan-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (11H) (80 mg, yield 26.6%).

[0592] LC-MS, M / Z (ESI): 292.6 [M+H] + .

[0593] Step 8: Synthesis of 2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-3-methylbenzamide (I-11)

[0594]

[0595] At room temperature, 4-(6-amino-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (80 mg, 0.27 mmol) was added to 1 mL of DMF, followed by DIPEA (70.9 mg, 0.55 mmol), HATU (156.7 mg, 0.41 mmol), and 2-fluoro-3-methylbenzoic acid (63.5 mg, 0.41 mmol). The mixture was stirred at room temperature for 16 h. Add water (10 ml), extract with DCM (10 ml × 3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate, and extract by thin-layer chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid 2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-3-methylbenzamide (I-11) (33 mg, yield 28.1%).

[0596] 1H NMR(400MHz,DMSO-d6)δ11.4(s,1H),8.47(s,1H),7.70(t,1H),7.52(d,1H),7.50(s,1H),7.28(t,1H),6 .15(s,1H),4.51(s,1H),3.75(s,3H),2.97(t,2H),2.33(d,3H),1.88(t,2H),1.60(t,6H),1.18(s,6H).

[0597] LC-MS, M / Z (ESI): 428.6 [M+H] + .

[0598] Example 12: Preparation of target compound I-12

[0599] 3-Chloro-2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)benzamide (target compound I-12)

[0600]

[0601] The synthetic route for the target compound I-12 is shown below:

[0602]

[0603] Step 1: Synthesis of methyl 4-(methylamino)-3-nitrobenzene (12B)

[0604]

[0605] At room temperature, methyl 4-fluoro-3-nitrobenzene (40.0 g, 200 mmol) was added to 600 ml of THF, followed by an ethanol solution of methylamine (29%, 150 ml). The mixture was heated to 70 °C and stirred for 72 h. After cooling to room temperature, water (1500 ml) was added, and the mixture was extracted with DCM (1200 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give crude methyl 4-(methylamino)-3-nitrobenzene (12B) (30.0 g, yield 71.0%) as a pale yellow solid.

[0606] LC-MS, M / Z (ESI): 211.2 [M+H] + .

[0607] Step 2: Synthesis of methyl 3-amino-4-(methylamino)benzoate (12C)

[0608]

[0609] At room temperature, methyl 4-(methylamino)-3-nitrobenzene (25.0 g, 119 mmol) was added to 300 mL of methanol, followed by the addition of 10% Pd / C (2.5 g). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow solid methyl 3-amino-4-(methylamino)benzoate (12C) (18.0 g, yield 84.0%).

[0610] LC-MS, M / Z (ESI): 181.2 [M+H] + .

[0611] Step 3: Synthesis of methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (12D)

[0612]

[0613] At room temperature, methyl 3-amino-4-(methylamino)benzoate (18.0 g, 100 mmol) was added to 500 ml of DCM. The mixture was cooled to 0 °C, and methyl 4-chloro-4-oxobutyrate (18.0 g, 120 mmol) and triethylamine (30.0 g, 300 mmol) were added. The mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give a colorless crude solid methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (12D) (13.0 g, yield 66.7%).

[0614] LC-MS, M / Z (ESI): 295.6 [M+H] + .

[0615] Step 4: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (12E)

[0616]

[0617] Methyl 4-((2-amino-4-bromophenyl)(methyl)amine)-4-oxobutyrate (7.0 g, 23.8 mmol) was added to 300 mL of xylene at room temperature, followed by PPTS (1.8 g, 7.1 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (300 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (12E) (4.0 g, yield 60.8%).

[0618] LC-MS, M / Z (ESI): 277.4 [M+H] + .

[0619] Step 5: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (12F)

[0620]

[0621] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (4.0 g, 14.5 mmol) was added to 100 mL of concentrated sulfuric acid at room temperature, cooled to 0 °C, and potassium nitrate (2.9 g, 29 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (12F) (1.4 g, yield 30.1%).

[0622] LC-MS, M / Z (ESI): 322.4 [M+H] + .

[0623] Step 6: Synthesis of methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (12G)

[0624]

[0625] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (2.0 g, 6.2 mmol) was added to 50 mL of methanol at room temperature, followed by the addition of 10% Pd / C (200 mg). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:1) to give crude methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (12 g) (1.8 g, yield 99.2%).

[0626] LC-MS, M / Z (ESI): 292.6 [M+H] + .

[0627] Step 7: Synthesis of 4-(6-amino-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (12H)

[0628]

[0629] At room temperature, methyl 6-(2-fluorobenzamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazol-5-carboxylate (300 mg, 1.0 mmol) was added to 3 mL of dry THF. The mixture was cooled to 0 °C, and a lanthanum(III) chloride bis(lithium chloride) complex solution (0.6 mol / L, 1.7 mL) and methyl magnesium bromide (1.0 mol / L, 10.3 mL) were added. The mixture was stirred at room temperature for 16 h. 50 mL of saturated ammonium chloride solution was added, and the mixture was extracted with EA (80 mL × 3). The extract was dried, filtered, concentrated, and purified by thin-layer chromatography to obtain a white solid 4-(6-amino-5-(2-hydroxypropan-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (12H) (80 mg, yield 26.6%).

[0630] LC-MS, M / Z (ESI): 292.6 [M+H] + .

[0631] Step 8: Synthesis of 3-chloro-2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)benzamide (I-12)

[0632]

[0633] At room temperature, 4-(6-amino-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]indol-2-yl)-2-methylbutane-2-ol (80 mg, 0.27 mmol) was added to 1 ml of DMF, followed by DIPEA (70.9 mg, 0.55 mmol), HATU (156.7 mg, 0.41 mmol), and 2-fluoro-3-chlorobenzoic acid (71.9 mg, 0.41 mmol). The mixture was stirred at room temperature for 16 h. Add water (10 ml), extract with DCM (10 ml × 3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate, and pass through a thin-layer chromatography plate (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid 3-chloro-2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)benzamide (I-12) (35 mg, yield 28.4%).

[0634] 1H NMR(400MHz,DMSO-d6)δ11.5(s,1H),8.41(s,1H),7.84(d,1H),7.80(d,1H),7.50(s,1H),7.44(t ,1H),6.19(s,1H),4.46(s,1H),3.72(s,3H),2.92(t,2H),1.87(t,2H),1.60(t,6H),1.18(s,6H).

[0635] LC-MS, M / Z (ESI): 448.2 [M+H] + .

[0636] Example 13: Preparation of target compound I-13

[0637] 3-(difluoromethyl)-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-methylbenzamide (target compound I-13)

[0638]

[0639] The synthetic route for the target compound I-13 is shown below:

[0640]

[0641] Step 1: Synthesis of methyl 4-(methylamino)-3-nitrobenzene (13B)

[0642]

[0643] At room temperature, methyl 4-fluoro-3-nitrobenzene (40.0 g, 200 mmol) was added to 600 mL of THF, followed by an ethanol solution of methylamine (29%, 150 mL). The mixture was heated to 70 °C and stirred for 72 h. After cooling to room temperature, water (1500 mL) was added, and the mixture was extracted with DCM (1200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give crude methyl 4-(methylamino)-3-nitrobenzene (13B) (30.0 g, yield 71.0%) as a pale yellow solid.

[0644] LC-MS, M / Z (ESI): 211.2 [M+H] + .

[0645] Step 2: Synthesis of methyl 3-amino-4-(methylamino)benzoate (13C)

[0646]

[0647] At room temperature, methyl 4-(methylamino)-3-nitrobenzene (25.0 g, 119 mmol) was added to 300 mL of methanol, followed by the addition of 10% Pd / C (2.5 g). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow solid methyl 3-amino-4-(methylamino)benzoate (13C) (18.0 g, yield 84.0%).

[0648] LC-MS, M / Z (ESI): 181.2 [M+H] +

[0649] Step 3: Synthesis of methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (13D)

[0650]

[0651] At room temperature, methyl 3-amino-4-(methylamino)benzoate (18.0 g, 100 mmol) was added to 500 ml of DCM. The mixture was cooled to 0 °C, and methyl 4-chloro-4-oxobutyrate (18.0 g, 120 mmol) and triethylamine (30.0 g, 300 mmol) were added. The mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 5:1) to give a colorless crude solid methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (13D) (13.0 g, yield 66.7%).

[0652] LC-MS, M / Z (ESI): 295.6 [M+H] + .

[0653] Step 4: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]indole-5-carboxylate (13E)

[0654]

[0655] Methyl 3-amino-4-(4-methoxy-N-methyl-4-oxobutyramide)benzoate (7.0 g, 23.8 mmol) was added to 300 mL of xylene at room temperature, followed by PPTS (1.8 g, 7.1 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (500 mL) was added, and the mixture was extracted with EA (300 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (13E) (4.0 g, yield 60.8%).

[0656] LC-MS, M / Z (ESI): 277.4 [M+H] + .

[0657] Step 5: Synthesis of methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (13F)

[0658]

[0659] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (4.0 g, 14.5 mmol) was added to 100 mL of concentrated sulfuric acid at room temperature, cooled to 0 °C, and potassium nitrate (2.9 g, 29 mmol) was added. The mixture was stirred at low temperature for 2 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 3:1) to give a white solid methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (13F) (1.4 g, yield 30.1%).

[0660] LC-MS, M / Z (ESI): 322.4 [M+H] + .

[0661] Step 6: Synthesis of methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (13G)

[0662]

[0663] Methyl 2-(3-methoxy-3-oxopropyl)-1-methyl-6-nitro-1H-benzo[d]imidazolium-5-carboxylate (2.0 g, 6.2 mmol) was added to 50 mL of methanol at room temperature, followed by the addition of 10% Pd / C (200 mg). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:1) to give crude methyl 6-amino-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]imidazolium-5-carboxylate (13 g) (1.8 g, yield 99.2%).

[0664] LC-MS, M / Z (ESI): 292.6 [M+H] + .

[0665] Step 7: Synthesis of 4-(6-amino-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (13H)

[0666]

[0667] Methyl 6-(2-fluorobenzamide)-2-(3-methoxy-3-oxopropyl)-1-methyl-1H-benzo[d]indole-5-carboxylate (300 mg, 1.0 mmol) was added to 3 mL of dry THF at room temperature. The mixture was cooled to 0 °C, and a solution of lanthanum(III) chloride bis(lithium chloride) complex (0.6 mol / L, 1.7 mL) and methyl magnesium bromide (1.0 mol / L, 10.3 mL) were added. The mixture was stirred at room temperature for 16 h. 50 mL of saturated ammonium chloride solution was added, and the mixture was extracted with EA (80 mL × 3). The extract was dried, filtered, concentrated, and purified by thin-layer chromatography to obtain a white solid 4-(6-amino-5-(2-hydroxypropan-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (13H) (80 mg, yield 26.6%).

[0668] LC-MS, M / Z (ESI): 292.6 [M+H] + .

[0669] Step 8: Synthesis of 3-(difluoromethyl)-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-methylbenzamide (I-13)

[0670]

[0671] At room temperature, 4-(6-amino-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)-2-methylbutane-2-ol (150 mg, 0.52 mmol) was added to 5 ml of DMF, followed by DIPEA (134 mg, 1.0 mmol), HATU (296 mg, 0.78 mmol), and 3-(difluoromethyl)-2-methylbenzoic acid (144 mg, 0.77 mmol). The mixture was stirred at room temperature for 16 h. Add water (10 ml), extract with EA (10 ml × 3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate, and prepare a white solid 3-(difluoromethyl)-N-(2-(3-hydroxy-3-methylbutyl)-5-(2-hydroxypropane-2-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)-2-methylbenzamide (I-13) (30 mg, yield 12.6%) by thin-layer chromatography (petroleum ether: ethyl acetate (V / V) = 3:1).

[0672] 1H NMR(400MHz,DMSO-d6)δ11.3(s,1H),8.75(s,1H),7.71(t,2H),7.65(s,1H),7.53(d,1H),7.42(t,1H),6 .44(s,1H),4.50(s,1H),3.94(s,3H),3.20(t,2H),2.50(s,3H),1.91(t,2H),1.61(t,6H),1.20(s,6H).

[0673] LC-MS, M / Z (ESI): 460.6 [M+H] + .

[0674] Example 14: Preparation of target compound I-14

[0675] 2-Fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-methoxy-1-methyl-1H-benzo[d]imidazol-6-yl)-3-(trifluoromethyl)benzamide (target compound I-14)

[0676]

[0677] The synthetic route for the target compound I-14 is shown below:

[0678]

[0679] Step 1: Synthesis of 4-methoxy-N-methyl-2-nitroaniline (14B)

[0680]

[0681] At room temperature, 10.0 g (58.5 mmol) of 4-methoxy-1-fluoro-2-nitrobenzene was added to 50 mL of methylamine methanol solution (containing 7.3 g (233.9 mmol) of methylamine). The mixture was heated to 70 °C and stirred for 32 h. After cooling to room temperature, 100 mL of water was added, and the mixture was extracted with EA (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give crude 4-methoxy-N-methyl-2-nitrobenzene (14B) (10.0 g, yield 93.9%) as a pale yellow solid.

[0682] Step 2: 4-Methoxy-N 1 Synthesis of 1,2-methylphenyl-1,2-diamine (14C)

[0683]

[0684] At room temperature, 4-methoxy-N-methyl-2-nitroaniline (2 g, 11.0 mmol) was added to 100 mL of THF, followed by 100 mL of ethanol and 50 mL of water. Iron powder (2.5 g, 44.6 mmol) and ammonium chloride (2.5 g, 46.7 mmol) were also added. The mixture was heated to 90 °C and stirred for 2 h. After cooling to room temperature, the mixture was extracted with DCM (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a pale yellow solid, 4-methoxy-N-methyl-2-nitroaniline. 1 1,2-methylphenyl-1,2-diamine (14C) (1.5 g, yield 89.8%).

[0685] LC-MS, M / Z (ESI): 153.4 [M+H] + .

[0686] Step 3: Synthesis of methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (14D)

[0687]

[0688] The raw material 4-methoxy-N was prepared at room temperature. 1 1,2-methylphenyl-1,2-diamine (19.5 g, 128 mmol) was added to 1000 ml of DCM and cooled to 0 °C. Methyl 4-chloro-4-oxobutyrate (21 g, 139.5 mmol) and triethylamine (39.0 g, 386.1 mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (1500 ml) was added, and the mixture was extracted with DCM (500 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 5:1) to give a colorless crude solid, methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (14D) (18.0 g, yield 52.7%).

[0689] LC-MS, M / Z (ESI): 267.5 [M+H] +

[0690] Step 4: Synthesis of methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (14E)

[0691]

[0692] Methyl 4-((2-amino-4-methoxyphenyl)(methyl)amine)-4-oxobutyrate (18.0 g, 67.6 mmol) was added to 600 mL of xylene at room temperature, followed by PPTS (1.7 g, 6.8 mmol). The mixture was heated to 140 °C and stirred for 16 h. After cooling to room temperature, water (1000 mL) was added, and the mixture was extracted with EA (500 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (14E) (10.5 g, yield 62.5%) as a white solid.

[0693] LC-MS, M / Z (ESI): 249.4 [M+H] +

[0694] Step 5: Synthesis of methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (14F)

[0695]

[0696] Methyl 3-(5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (9.8 g, 39.5 mmol) was added to 50 mL of acetic acid at room temperature, cooled to 0 °C, and 50 mL of nitric acid was added. The mixture was stirred at low temperature for 1 h. A saturated sodium bicarbonate solution (500 mL) was added, and the mixture was extracted with EA (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (14F) (3.8 g, yield 32.8%).

[0697] LC-MS, M / Z (ESI): 294.5 [M+H] +

[0698] Step 6: Synthesis of methyl 3-(6-amino-5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (14G)

[0699]

[0700] Methyl 3-(5-methoxy-1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)propionate (1.2 g, 4.1 mmol) was added to 10 mL of methanol at room temperature, along with palladium on carbon (120 mg). H2 was bubbled through the mixture, and the mixture was stirred at room temperature for 16 h. The mixture was then filtered and concentrated to give crude white solid methyl 3-(6-amino-5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (14 g) (960 mg, yield 89.1%).

[0701] LC-MS, M / Z (ESI): 264.6 [M+H] + .

[0702] Step 7: Synthesis of methyl 3-(6-(2-fluoro-3-(trifluoromethyl)benzamide)-5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (14H)

[0703]

[0704] Methyl 3-(6-amino-5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (300 mg, 1.14 mmol) was added to 5 mL of DMF at room temperature, along with HATU (650 mg, 1.7 mmol) and DIPEA (294 mg, 2.28 mmol), and 2-fluoro-3-(trifluoromethyl)benzoic acid (237 mg, 1.14 mmol). The mixture was stirred at room temperature for 16 h. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 3:1) to give a white solid methyl 3-(6-(2-fluoro-3-(trifluoromethyl)benzamide)-5-methoxy-1-methyl-1H-benzo[d]imidazol-2-yl)propionate (14H) (380 mg, yield 73.5%).

[0705] LC-MS, M / Z (ESI): 454.3 [M+H] + .

[0706] Step 8: Synthesis of 2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-methoxy-1-methyl-1H-benzo[d]imidazol-6-yl)-3-(trifluoromethyl)benzamide (I-14)

[0707]

[0708] Methyl 3-(6-(2-fluoro-3-(trifluoromethyl)benzamide)-5-methoxy-1-methyl-1H-benzo[d]indol-2-yl)propionate (180 mg, 0.39 mmol) was added to 5 ml of dry THF at room temperature, cooled to 0 °C, and lithium chloride (334 mg, 7.9 mol) and methyl magnesium bromide (1.0 mol / L, 4 ml) were added. The mixture was stirred at room temperature for 24 h. Add 5 ml of water, adjust pH to 6 with 1 mol / L hydrochloric acid, concentrate, and prepare the separation method as follows (chromatographic column: Welch, Ultimate C18 column, 10 μm, 21.2 mm × 250 mm. Mobile phase A: 1‰ trifluoroacetic acid pure water solution, mobile phase B is acetonitrile solution. Gradient conditions: 0-3 min, mobile phase A is maintained at 90%, 3-18 min gradient elution, from 90% to 5%, 18-22 min maintain 5%), to prepare a white solid 2-fluoro-N-(2-(3-hydroxy-3-methylbutyl)-5-methoxy-1-methyl-1H-benzo[d]imidazol-6-yl)-3-(trifluoromethyl)benzamide (I-14) (51 mg, yield 28.3%).

[0709] 1H NMR(400MHz,DMSO-d6)δ9.85(d,1H),8.17(s,1H),8.09(t,1H),7.98(t,1H),7.57(t,1H),7 .33(s,1H),4.51(s,1H),3.87(s,3H),3.74(s,3H),2.96(t,2H),1.85(t,2H),1.17(s,6H).

[0710] LC-MS, M / Z (ESI): 454.3 [M+H] + .

[0711] Test A: Anti-inflammatory activity test

[0712] Test Example A-1: ​​Inhibitory activity of the compound against IRAK4 kinase

[0713] First, the test compound was dissolved in DMSO to prepare a 10 mM stock solution. Then, 1× kinase base buffer (50 mM HEPES, pH 7.5; 0.0015% Brij-35) and stop buffer (100 mM HEPES, pH 7.5; 0.015% Brij-35, 0.2% Coating Reagent#3, 50 mM EDTA) were prepared for use.

[0714] Next, the stock solution of the compound was diluted 50-fold with DMSO to the highest concentration gradient. 100 μL of this diluted solution was transferred to the wells of a 96-well plate, and then serially diluted 4-fold until 10 concentration gradients were achieved. 100 μL of DMSO was added to two empty wells of the same 96-well plate as a compound-free control and an enzyme-free control, serving as the source plate. 10 μL of the compound was transferred to a new 96-well plate as an intermediate plate.

[0715] Add 90 μL of 1× kinase buffer to each well of the intermediate plate and incubate on a shaker for 10 min to mix thoroughly. Transfer 5 μL from each well to two replicates in a 384-well plate to create the test plate.

[0716] A 2.5× enzyme solution was prepared by adding IRAK4 kinase (Carna) to 1× kinase base buffer. A 2.5× peptide solution was prepared by adding FAM-labeled peptide and ATP to 1× kinase base buffer. 10 μL of the 2.5× enzyme solution was added to each well of a 384-well plate containing 5 μL of the compound, and incubated at room temperature for 10 min. 10 μL of the 2.5× peptide solution was added to each well of a 384-well test plate, and incubated at 28°C for one hour. After incubation, 30 μL of stop buffer was added to each well to stop the reaction. Data from each well were then collected using a Caliper instrument, and the IC50 value for each compound was calculated after curve fitting.

[0717] Table 1: Inhibitory activity of the tested compounds against IRAK4 kinase

[0718]

[0719]

[0720] The experimental results show that all compounds of the present invention have good inhibitory effects on IRAK4 kinase, especially compounds I-1, I-2, I-3, I-4, I-5 and I-6, which have significant inhibitory effects on IRAK4 kinase, indicating that the compounds of the present invention have the potential to resist inflammatory responses.

[0721] Test Example A-2: Inhibitory activity of the compound on R848-stimulated PBMC production of TNF-α

[0722] First, thaw the frozen PBMC cells, centrifuge at 200 rpm for 5 min, collect the supernatant, resuspend in 1640 + 10% FBS + 1% P / S complete medium, and count the cells. Seed the cells at a density of 8 × 10⁴ cells / well, 100 μl / well, into 96-well plates and grow overnight.

[0723] Next, a 10 mM stock solution of the compound was prepared and serially diluted 3-fold with DMSO. Then, the concentrations were diluted 500-fold with 1640 + 10% FBS + 1% P / S complete medium to achieve a final concentration of 4-fold (20 μM). The prepared compound was added to cells at 50 μl / well, with a control well containing no compound, and pre-incubated for 30 min. Simultaneously, a 10 mg / ml stock solution of R848 was prepared and diluted 100-fold and then 31.25-fold with 1640 + 10% FBS + 1% P / S medium to achieve a final concentration of 4-fold (3.2 μg / ml). The prepared R848 was added to cells at 50 μl / well, with a control well containing no R848. After 24 h of incubation, the supernatant was collected, and 50% of the supernatant was analyzed using a Human TNF-α assay kit (Invitrogen, LOT: 189974010).

[0724] Table 2: Inhibitory activity of the tested compounds on R848-stimulated PBMC production of TNF-α

[0725] compound IC50(nM) I-1 13 I-2 10 I-3 8.6 I-4 220 I-5 127 I-6 66.3 I-9 121.9

[0726] The experimental results show that all the compounds of the present invention have good inhibitory activity against TNF-α production. In particular, compounds I-1, I-2, and I-3 have significant inhibitory activity against TNF-α production, indicating that the compounds of the present invention have the potential to resist inflammatory responses.

[0727] Test Example A-3: Inhibitory activity of the compound on R848-stimulated production of IL6 in human whole blood

[0728] 30ml of venous blood was drawn from a healthy volunteer into a heparin anticoagulant tube.

[0729] Prepare a 100× compound DMSO solution with an initial concentration of 500 μM, and dilute it 3-fold to 12 concentration gradients (including two 0 points);

[0730] Add 2 μl of 100× positive compound DMSO solution at different concentration gradients to each well of a 96-well round-bottom plate, followed by 200 μl of heparin-anticoagulated whole blood. Mix until the final concentration of the positive compound is 1× working concentration. Set up three replicates for each compound. The final concentration of DMSO is 1%. Cover the plate and incubate at 37°C for 30 min.

[0731] Take an appropriate amount of 10 mg / ml R848 stock solution (DMSO) and dilute it with pure water to 5 μg / ml (20×). Add 10 μl of R848 aqueous solution (5 μg / ml) to each well to make the final concentration of R848 0.25 μg / ml. Add only 10 μl of pure water to one of the 0-point concentration wells as a non-irritating well. After mixing, cover the plate and incubate at 37°C for 4 hours.

[0732] After incubation, the plasma was centrifuged at 1500 rpm for 10 min, and 25 μl of plasma was collected. The concentration of human IL6 was detected using an ELISA kit (Biolegend, Cat:430504).

[0733] The experimental results show that all compounds of the present invention have a good inhibitory effect on the production of IL-6, indicating that the compounds of the present invention have the potential to resist inflammatory responses.

[0734] Test B: Anticancer activity test

[0735] Test Example B-1: Inhibitory activity of the compound against FLT3-WT, FLT3-ITD, or FLT3-D35Y kinases

[0736] First, the test compound was dissolved in DMSO to prepare a 10 mM stock solution. Then, 1× kinase base buffer (50 mM HEPES, pH 7.5; 0.0015% Brij-35) and stop buffer (100 mM HEPES, pH 7.5; 0.015% Brij-35, 0.2% Coating Reagent#3, 50 mM EDTA) were prepared for use.

[0737] Next, the stock solution of the compound was diluted 50-fold with DMSO to the highest concentration gradient. 100 μL of this diluted solution was transferred to the wells of a 96-well plate, and then serially diluted 4-fold until 10 concentration gradients were achieved. 100 μL of DMSO was added to two empty wells of the same 96-well plate as a compound-free control and an enzyme-free control, serving as the source plate. 10 μL of the compound was transferred to a new 96-well plate as an intermediate plate.

[0738] Add 90 μL of 1× kinase buffer to each well of the intermediate plate and incubate on a shaker for 10 min to mix thoroughly. Transfer 5 μL from each well to two replicates in a 384-well plate to create the test plate.

[0739] A 2.5× enzyme solution was prepared by adding the corresponding kinase (FLT3-WT / FLT3-ITD / FLT3-835Y, Carna) to 1× kinase base buffer. A 2.5× peptide solution was prepared by adding FAM-labeled peptides and ATP to the same 1× kinase base buffer. 10 μL of the 2.5× enzyme solution was added to each well of a 384-well plate containing 5 μL of the compound, and incubated at room temperature for 10 min. 10 μL of the 2.5× peptide solution was added to each well of a 384-well test plate, and incubated at 28°C for 0.5 h. After incubation, 30 μL of stop buffer was added to each well to stop the reaction. Data from each well were then collected using a Caliper instrument, and the IC50 value for each compound was calculated after curve fitting.

[0740] Table 3: Inhibitory activity of FLT3-WT, FLT3-ITD, or FLT3-D35Y kinases

[0741]

[0742] The experimental results show that compounds I-1 and I-2 of the present invention also have significant inhibitory effects on wild-type, ITD mutant and D835Y mutant FLT3 kinases, indicating that the compounds have the potential to combat FLT3 mutant cancers.

[0743] Test Example B-2: CTG Method for Detecting OCI-LY10 Tumor Cell Proliferation

[0744] The OCI-LY10 (human B-cell lymphoma) cell line was routinely passaged in a 37°C, 5% CO2 culture system. Cells growing in the exponential growth phase were harvested and counted for plate seeding.

[0745] Cells were counted using a trypan blue-stained hemocytometer, and the cell concentration was adjusted to an appropriate density. Cells were then seeded into 96-well plates with 90 μL of cell suspension in each well. Wells containing only culture medium were included as blank controls. The plates were incubated overnight at 37°C in a 5% CO2 system to create cell assay plates.

[0746] The compound was diluted with DMSO in a 96-well plate until different gradient concentrations were reached.

[0747] Next, prepare a V-bottom cell culture plate. Add 198 μL of test culture medium and 2 μL of diluted compound solution to each well, and add 2 μL of DMSO to the control well. Mix well by pipetting. This V-bottom culture plate serves as a 10× concentration compound plate.

[0748] From each well of the compound plate, aspirate 10 μL of the compound-culture medium mixture or the mixture from the control well and transfer it to the cell assay plate. The final concentration of DMSO is 0.1%. Return the cell assay plate to the culture system and incubate for 3 days to allow cell growth.

[0749] Cell samples were treated with Promega's CellTiter-Glo kit (Promega-G7573), and luminescence signals were recorded on a 2104 EnVision plate reader. The inhibition rate (IR) at different doses of the test compound was calculated: IR (%) = (1 – (RLU compound – RLU blank) / (RLU control – RLU blank)) × 100%, and then the IC50 value was calculated.

[0750] Table 4: Tumor cell inhibitory activity

[0751] compound IC50 (μM) I-2 0.29 I-5 5.1 I-9 89

[0752] The experimental results show that all compounds of the present invention exhibit strong inhibitory activity against the proliferation of OCI-LY10 cells, especially compounds I-2 and I-5, which show significant inhibitory activity against the proliferation of OCI-LY10 cells, indicating that the compounds of the present invention have the potential to combat cancer.

[0753] Test C: Pharmacokinetic Study

[0754] Pharmacokinetic studies were conducted in mice using male ICR mice (20-25g, fasted overnight). Three mice were administered 10mg / kg orally via gavage. Blood samples were collected before administration and at 15, 30 minutes, 1, 2, 4, 8, and 24 hours after administration. 6800g of blood samples were centrifuged at 2-8℃ for 6 minutes, and plasma was collected and stored at -80℃. Plasma samples at each time point were mixed with 3-5 times the volume of acetonitrile solution containing internal standard, vortexed for 1 minute, centrifuged at 13000 rpm at 4℃ for 10 minutes, and the supernatant was mixed with 3 times the volume of water. An appropriate amount of the mixture was analyzed by LC-MS / MS. Key pharmacokinetic parameters were analyzed using a non-compartmental model in WinNonlin 7.0 software.

[0755] Pharmacokinetic studies were conducted on male SD rats (180-240g) that had been fasted overnight. Three rats were administered the drug 10 mg / kg orally via gavage. Blood samples were collected before administration and at 15, 30 minutes, 1, 2, 4, 8, and 24 hours after administration. Blood samples were centrifuged at 8000 rpm for 6 minutes at 4°C, and plasma was collected and stored at -20°C. Plasma samples at each time point were mixed with 3-5 times the volume of acetonitrile solution containing an internal standard, vortexed for 1 minute, centrifuged at 13000 rpm for 10 minutes at 4°C, and the supernatant was mixed with 3 times the volume of water. An appropriate amount of the mixture was analyzed by LC-MS / MS. The main pharmacokinetic parameters were analyzed using a non-compartmental model with WinNonlin 7.0 software.

[0756] Canine pharmacokinetic studies were conducted using male beagle dogs, weighing 8-10 kg, which were fasted overnight. Three beagle dogs were administered 5 mg / kg orally via gavage. Blood samples were collected before administration and at 15, 30 minutes, and 1, 2, 4, 8, and 24 hours after administration. Blood samples were centrifuged at 8000 rpm for 6 minutes at 4°C, and plasma was collected and stored at -20°C. Plasma samples at each time point were mixed with 3-5 times the volume of acetonitrile solution containing internal standard, vortexed for 1 minute, centrifuged at 13000 rpm for 10 minutes at 4°C, and the supernatant was mixed with 3 times the volume of water. An appropriate amount of the mixture was analyzed by LC-MS / MS. The main pharmacokinetic parameters were analyzed using a non-compartmental model in WinNonlin 7.0 software.

[0757] Table 5: Results of Pharmacokinetic Studies in Mice

[0758]

[0759] The results of the pharmacokinetic studies in mice showed that the compound of the present invention exhibited excellent pharmacokinetic properties, high exposure levels, and good drug-likeness.

[0760] Table 6: Results of Rat Pharmacokinetic Study

[0761]

[0762] The results of the pharmacokinetic studies in rats showed that the compound of the present invention exhibited excellent pharmacokinetic properties, high exposure levels, and good drug-likeness.

[0763] Table 7: Results of canine pharmacokinetics studies

[0764]

[0765] The pharmacokinetic results in dogs show that the compound of this invention exhibits excellent pharmacokinetic properties, high exposure levels, and good drug-likeness.

Claims

1. The compound represented by Formula I, its stereoisomers, or pharmaceutically acceptable salts: in, L is absent or -0-(CH2) m - and m is 0 or 1 ; R 1 Selected from unsubstituted or R a The substituted 4-8 membered heterocyclic alkyl group is oxetane, oxetane, oxetane, oxetane, azirbutane, aziretane, or morpholinyl; the R-substituted... a In the substituted 4-8 membered heterocyclic alkyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, amino, C1-C3 alkyl; when there are multiple substituents, the substituents may be the same or different; When L is -O-(CH2) m -time,-LR 1 For -O-(CH2) m -R 1 ; Ring A is selected from 5-8-membered heteroaryl groups, wherein the 5-8-membered heteroaryl group is pyrrole, pyrazole, triazole, furan, oxazole, thiophene, thiazole, pyridine, pyrazine, or pyrimidine; n is 1 or 2; R 2 Selected from halogens, hydroxyl groups, cyano groups, amino groups, C1-C3 alkyl groups, C1-C3 alkyl groups substituted with 1, 2, or 3 of the same or different halogens, unsubstituted or substituted with R b Substituted 5-8 membered heteroaryl groups; the R-substituted heteroaryl group b In the substituted 5-8 membered heteroaryl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C3 alkyl, or C1-C3 alkyl substituted with 1, 2, or 3 identical or different halogens; when n is not 1, R 2 Independently, they can be either the same or different; When R 2 For not replaced or by R b The substituted 5-8 heteroaryl group is pyrrole, pyrazole, triazole, furan, oxazole, thiophene, thiazole, pyridine, pyrazine or pyrimidine.

2. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 1 For R a When substituted with 4-8 membered heterocyclic alkyl groups, the number of substituted groups is independently 1-3.

3. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, R a It is a hydroxyl group.

4. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R a When the halogen is halogen, the halogen is F, Cl, Br, or I.

5. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 1 For R a When substituted with a 4-8 membered heterocyclic alkyl group, the number of substituted groups is independently 1.

6. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R a When the halogen is halogen, the halogen is F or Cl.

7. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R a When the alkyl group is C1-C3, the C1-C3 alkyl group is methyl, ethyl, n-propyl, or isopropyl.

8. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 For R b When the 5-8 membered heteroaryl group is substituted, the number of substituted groups is independently 1-3.

9. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 When the halogen is halogen, the halogen is F, Cl, Br, or I.

10. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 When the halogen is a C1-C3 alkyl group substituted with 1, 2 or 3 identical or different halogens, the halogen is F, Cl, Br or I.

11. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R b When the halogen is halogen, the halogen is F, Cl, Br, or I.

12. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R b When the halogen is a C1-C3 alkyl group substituted with 1, 2 or 3 identical or different halogens, the halogen is F, Cl, Br or I.

13. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When ring A is selected from 5-8 heteroaryl groups, the 5-8 heteroaryl group is an oxazole or a pyridine.

14. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 For not replaced or by R b The substituted 5-8 heteroaryl group is a pyrazole or pyridine.

15. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 For R b When 5-8 heteroaryl groups are substituted, the number of substitutions is independently 1 or 2.

16. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 When the halogen is halogen, the halogen is F or Cl.

17. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 When the alkyl group is C1-C3, the C1-C3 alkyl group is methyl, ethyl, n-propyl, or isopropyl.

18. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 When the C1-C3 alkyl group is substituted with 1, 2 or 3 of the same or different halogens, the C1-C3 alkyl group is methyl, ethyl, n-propyl or isopropyl.

19. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R 2 When the halogen is a C1-C3 alkyl group substituted with 1, 2 or 3 identical or different halogens, the halogen is F or Cl.

20. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R b When the alkyl group is C1-C3, the C1-C3 alkyl group is methyl, ethyl, n-propyl, or isopropyl.

21. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R b When the C1-C3 alkyl group is substituted with 1, 2 or 3 of the same or different halogens, the C1-C3 alkyl group is methyl, ethyl, n-propyl or isopropyl.

22. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, When R b When the halogen is a C1-C3 alkyl group substituted with 1, 2 or 3 identical or different halogens, the halogen is F or Cl.

23. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, for , , ; or, for , , .

24. The compound of formula I according to claim 1, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, It is either Option 1 or Option 3; among them, Option 1 L represents non-existence or -O-(CH2). m - where m is 0 or 1; R 1 For not replaced or by R a The substituted 4-8 membered heterocyclic alkyl group is oxetane, oxetane, oxetane, oxetane, azirbutane, aziretane, or morpholino; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, amino, C1-C3 alkyl; when there are multiple substituents, the substituents may be the same or different; When L is -O-(CH2) m -time,-LR 1 For -O-(CH2) m -R 1 ; n is 1 or 2; R 2 Selected from unsubstituted or R b Substituted 5-8 membered heteroaryl groups; the R-substituted heteroaryl group b In the substituted 5-8 membered heteroaryl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, cyano, amino, C1-C3 alkyl, or C1-C3 alkyl substituted with 1, 2, or 3 identical or different halogens; when n is not 1, R 2 Independently, they can be either the same or different; When R 2 For not replaced or by R b The substituted 5-8-membered heteroaryl group is pyrrole, pyrazole, triazole, furan, oxazole, thiophene, thiazole, pyridine, pyrazine or pyrimidine; Option 3 L represents non-existence or -O-(CH2). m - where m is 0 or 1; R 1 For not replaced or by R a The substituted 4-8 membered heterocyclic alkyl group is oxetane, oxetane, oxetane, oxetane, azirbutane, aziretane, or morpholinyl; the R-substituted... a In the substituted 4-8 membered heterocyclic alkyl groups, each substitution independently refers to one or more of the following substituents: halogen, hydroxyl, amino, C1-C3 alkyl; when there are multiple substituents, the substituents may be the same or different; When L is -O-(CH2) m -time,-LR 1 For -O-(CH2) m -R 1 ; n is 1 or 2; R 2 Selected from halogens, hydroxyl groups, cyano groups, amino groups, C1-C3 alkyl groups, and C1-C3 alkyl groups substituted with 1, 2, or 3 of the same or different halogens; when n is not 1, R 2 Independently, they can be the same or different.

25. The compound of formula I according to claim 24, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, In scheme 1, L represents either non-existent or -O-(CH2). m - where m is 0 or 1; R 1 For not replaced or by R a The substituted azircyclopentane, oxacyclopentane, oxacyclohexane, azircyclobutane, oxacyclobutane, azircyclohexane, or morpholinoyl; each of the substituted substances independently refers to one or more hydroxyl substitutions; n is 1 or 2; R 2 Selected from unsubstituted or R b The substituted pyridine, pyrazole, triazole, furan, oxazole, thiophene, thiazole, pyrrole, pyrazine, or pyrimidine, wherein each substitution independently refers to one or more C1-C3 alkyl substitutions; when n is not 1, R 2 Independently, they can be either the same or different; In scheme 3, L represents either non-existent or -O-(CH2). m - where m is 0 or 1; R 1 For not replaced or by R a Substituted azircyclopentane, oxacyclopentane, oxacyclohexane, azircyclobutane, oxacyclobutane, azircyclohexane, or morpholinoyl, wherein the R a In the substituted 4-6 membered heterocyclic alkyl groups, each substitution independently refers to one or more hydroxyl substitutions, and when there are multiple substituents, the substituents may be the same or different; n is 1 or 2; R 2 Selected from C1-C3 alkyl groups substituted with 1, 2, or 3 identical or different halogens; when n is not 1, R 2 Independently, they can be the same or different.

26. The compound represented by Formula I, its stereoisomers, or pharmaceutically acceptable salts, characterized in that, The compound represented by Formula I is selected from any of the following compounds: 。 27. A pharmaceutical composition, characterized in that, It comprises a compound of Formula I as described in any one of claims 1-26, its stereoisomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable excipient.

28. Use of the compound of Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 27, as described in any one of claims 1-26, in the preparation of a medicament for treating diseases associated with IRAK4.

29. The use according to claim 28, characterized in that, The IRAK4-related diseases include autoimmune diseases and cancer.

30. The use according to claim 29, characterized in that, The autoimmune diseases mentioned are multiple sclerosis, systemic lupus erythematosus, psoriasis, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis, reactive arthritis, systemic juvenile idiopathic arthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, and allergic eczema.

31. The use according to claim 29, characterized in that, The cancer in question is a tumor in the head.

32. The use according to claim 29, characterized in that, The cancer in question is a hematologic malignancy.

33. The use according to claim 29, characterized in that, The cancer in question is leukemia.

34. The use according to claim 29, characterized in that, The cancers mentioned include brain cancer, kidney cancer, liver cancer, stomach cancer, vaginal cancer, ovarian cancer, gastric tumors, breast cancer, cystocolic cancer, prostate cancer, pancreatic cancer, lung cancer, cervical cancer, testicular cancer, skin cancer, bone cancer, thyroid cancer, sarcoma, glioblastoma, neuroblastoma, multiple myeloma, gastrointestinal cancer, neck tumors, adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, non-small cell lung cancer, Hodgkin's and non-Hodgkin's lymphoma, breast cancer, follicular carcinoma, papillary carcinoma, seminoma, and melanoma. Myeloma, acute myeloid leukemia, chronic myeloid leukemia, diffuse large B-cell lymphoma, activated B-cell-like diffuse large B-cell lymphoma, chronic lymphocytic leukemia, chronic lymphocytic lymphoma, primary exudative lymphoma, Burkitt lymphoma, acute lymphocytic leukemia, B-cell prelymphocytic leukemia, lymphoplasmacytic lymphoma, Waldenström macroglobulinemia, splenic marginal zone lymphoma, intravascular large B-cell lymphoma, plasmacytoma, and multiple myeloma.

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