Benzimidazo 2-amino-1, 3, 4-thiadiazole derivative as well as preparation method and pharmaceutical application thereof

Abstract

The invention belongs to the technical field of medicines, and relates to a benzimidazo 2-amino-1, 3, 4-thiadiazole compound and a pharmaceutical application thereof, in particular to a benzimidazo 2-amino-1, 3, 4-thiadiazole derivative shown as a formula I, a pharmaceutical salt thereof, a preparation method of the benzimidazo 2-amino-1, 3, 4-thiadiazole derivative, a composition containing one or more of the compounds, and a pharmaceutical application of the benzimidazo 2-amino-1, 3, 4-thiadiazole compound. The invention also discloses application of the compound as an STAT3 signal channel inhibitor in the aspect of treating tumor diseases. # imgabs0 #

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to compounds that inhibit human tumor cells and tumor immunotherapy. The invention also relates to the synthesis of such compounds and their use as effective drug candidates for the clinical treatment of cancer patients. More specifically, the invention relates to certain benzimidazolo-2-amino-1,3,4-thiadiazole derivatives that inhibit tumor cells with high STAT3 expression, and their use in the preparation of drugs for the prevention, alleviation, and / or treatment of anti-tumor effects. Background Art

[0002] Signal transducer and activator of transcription protein 3 (STAT3) is a transcription factor that can transduce extracellular signals to the cell nucleus and is an important member of the STATs family. STAT3 protein is a multi-domain protein consisting of an N-terminal domain, a coiled-coil domain, a DNA binding domain, a linker domain, an SH2 domain, and a C-terminal transcription activation domain. Among them, Tyr705 in the SH2 domain is crucial for the activation of STAT3. When Tyr705 is phosphorylated, STAT3 protein dimerizes, and then the activity of STAT3 protein is activated, thereby playing a role in transduction and transcription (Eur. J. Med. Chem., 2020 (190), 112122).

[0003] STAT3 is closely related to the survival, proliferation, invasion, metastasis, angiogenesis and immune escape of tumor cells (Cell Prolif., 2021, 54: e12974). Abnormal activation of STAT3 often occurs in various solid tumors and hematological malignancies. Abnormal expression of STAT3 has been observed in breast cancer, ovarian cancer, melanoma, prostate cancer, pancreatic cancer, multiple myeloma, leukemia, hematopoietic cancer, etc. Therefore, inhibiting STAT3 can block the occurrence of cancer in the early stages of cancer development (Eur. J. Med. Chem., 2020 (187), 111922).

[0004] Depending on the region of action, small molecule compounds that target the STAT3 protein can be divided into inhibitors targeting the coiled-coil domain, the DNA binding domain, the linker domain, and the SH2 domain. Among them, small molecule inhibitors targeting the STAT3 SH2 domain, as the current research focus of direct STAT3 inhibitors, have the advantages of high selectivity, strong binding affinity, and a clear target of action, and are of great significance in cancer treatment (Eur. J. Med. Chem., 2019, 1119-22). Small molecule inhibitors targeting the STAT3 SH2 domain mainly inhibit STAT3 function by interfering with the binding site of STAT3 protein dimerization, inhibiting STAT3 dimerization and the formation of STAT3 dimer-DNA complexes.

[0005] The design idea of ​​this patent is to combine the anti-tumor skeleton benzimidazole and the active fragment 2-amino-1,3,4-thiadiazole through the principle of advantageous skeleton splicing to obtain benzimidazole-2-amino-1,3,4-thiadiazole derivatives, in order to discover new STAT3-targeted anti-tumor compounds with high efficiency, low toxicity, suitable physicochemical properties and strong specificity.

[0006] The benzimidazolo 2-amino-1,3,4-thiadiazole compounds of the general formula I and their derivatives are new compounds and have no application or related reports in preventing and treating cancer and its complications. Summary of the Invention

[0007] One aspect of the present invention is a compound having the following structure (Formula I) and various intermediates and by-products involved in the synthesis thereof, and optical isomers, pharmaceutically acceptable salts, or solvates thereof:

[0008]

[0009] Where:

[0010] R1 is fluorine, chlorine, bromine, iodine, straight chain or branched C 1-6 Alkoxy, substituted and unsubstituted C 6-12 Phenoxy, substituted and unsubstituted N-heterocyclylamino, wherein the phenoloxy and N-heterocyclylamino groups can be unsubstituted or substituted by one or more groups selected from the following groups, these groups include straight chain or branched C 1-6 Alkyl, linear or branched C 1-6 Alkoxy, linear or branched amino, halogen, carboxyl, C 1-6 wherein the heterocyclic groups may be unsubstituted or substituted by one or more groups selected from the following groups, including C 1-6 Alkyl, C 1-6 Alkoxy, amino, halogen.

[0011] R2 is hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy substituted C 1-6 Alkyl, alkyl-substituted C 1-6 Alkyl and C 3-6 Cycloalkyl, wherein the cycloalkyl may be unsubstituted or substituted by one or more groups selected from the following groups, these groups include C 1-6 Alkyl, C 1-6 Alkoxy, amino.

[0012] R3 is hydrogen, substituted and unsubstituted heteroaryl, substituted and unsubstituted aryl, wherein the aryl or heteroaryl can be unsubstituted or substituted by one or more groups selected from the following groups, including C 1-6 Alkyl, C 1-6 Alkoxy, amino, halogen, cyano, C 6-12 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, C 1-6 Ester group, hydroxyl group, C 1-6 Amide group, methylsulfone group.

[0013] R4 is hydrogen, C 1-6 Alkyl, substituted and unsubstituted heteroaryl, substituted and unsubstituted aryl, wherein the aryl or heteroaryl can be unsubstituted or substituted by one or more groups selected from the following groups, these groups include C 1-6 Alkyl, C 1-6 Alkoxy, linear or branched substituted and unsubstituted amino, C 1-12 Cyclic amino, halogen, cyano, C 6-12 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, nitro, C 1-6 Ester group, hydroxyl group, C 1-6 Amide, C 1-6 Heteroaryl, C 4-12 Triazole, trifluoromethylphenyloxy, C 1-12 Sulfanyl, C 1-12 Sulfonamide, phenyl, morpholinyl, C 1-12 Alkynyl, C 1-6 Cycloalkoxy or C 1-12 Alkyleneoxy, hydrazine, C 1-12 Hydrazide group, C 5-12 Aromatic heterocycle.

[0014] Preferred

[0015] R1 is fluorine, chlorine, bromine, iodine, straight chain or branched C 1-4 Alkoxy, substituted and unsubstituted phenoxy, substituted and unsubstituted naphthoxy, substituted or unsubstituted piperazinyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include straight chain or branched C1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, carboxyl, C 1-4 ester group and hydroxyl group.

[0016] R2 is hydrogen, C 1-4 Alkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, alkyl-substituted C 1-4 Alkyl and C 3-6 Cycloalkyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino.

[0017] R3 is hydrogen, C 1-4 alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted naphthyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted pyrazinyl, substituted and unsubstituted pyridazinyl, substituted and unsubstituted benzimidazolyl, substituted and unsubstituted benzoxazolyl, substituted and unsubstituted benzothiazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, cyano, phenoxy, naphthoxy, trifluoromethyl, trifluoromethoxy, carboxyl, C 1-4 Ester group, hydroxyl group, C 1-4 Amide group, methyl sulfone group.

[0018] R4 is hydrogen, C 1-4 alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted naphthyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted pyrazinyl, substituted and unsubstituted pyridazinyl, substituted and unsubstituted benzimidazolyl, substituted and unsubstituted benzoxazolyl, substituted and unsubstituted benzothiazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, linear or branched substituted and unsubstituted amino, C 1-10 Cyclic amino, halogen, cyano, C 6-10 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, nitro, C 1-4Ester group, hydroxyl group, C 1-4 Amide, C 1-4 Heteroaryl, C 4-10 Triazole, trifluoromethylphenyloxy, C 1-10 Sulfanyl, C 1-10 Sulfonamide, phenyl, morpholinyl, C 1-10 Alkynyl, C 1-4 Cycloalkoxy or C 1-10 Alkyleneoxy, hydrazine, C 1-10 Hydrazide group, C 5-10 Aromatic heterocycle.

[0019] More preferred

[0020] R1 is fluorine, chlorine, bromine, iodine, straight chain or branched C 1-4 Alkoxy, substituted and unsubstituted phenoxy, substituted and unsubstituted naphthoxy, substituted or unsubstituted piperazinyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include straight chain or branched C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, carboxyl, C 1-4 Ester group and hydroxyl group;

[0021] R2 is hydrogen, C 1-4 Alkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, alkyl-substituted C 1-4 Alkyl and C 3-6 Cycloalkyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino.

[0022] R3 is hydrogen, C 1-4 alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted benzimidazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, cyano, phenoxy, naphthoxy, trifluoromethyl, trifluoromethoxy, carboxyl, C 1-4 Ester group, hydroxyl group, C 1-4 Amide group, methyl sulfone group.

[0023] R4 is hydrogen, C 1-4alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted benzimidazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, linear or branched substituted and unsubstituted amino, C 1-10 Cyclic amino, halogen, cyano, C 6-10 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, nitro, C 1-4 Ester group, hydroxyl group, C 1-4 Amide, C 1-4 Heteroaryl, C 4-10 Triazole, trifluoromethylphenyloxy, C 1-10 Sulfanyl, C 1-10 Sulfonamide, phenyl, morpholinyl, C 1-10 Alkynyl, C 1-4 Cycloalkoxy or C 1-10 Alkyleneoxy, hydrazine, C 1-10 Hydrazide group, C 5-10 Aromatic heterocycle.

[0024] In addition, the present invention also relates to the use of a pharmaceutically effective amount of a compound of formula I or its stereoisomers, pharmaceutically acceptable salts, or prodrugs for the preparation of a drug for preventing or treating tumor-related diseases.

[0025] The most preferred compounds include but are not limited to

[0026] Compound 1: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0027]

[0028] Compound 2: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0029]

[0030] Compound 3: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0031]

[0032] Compound 4: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0033]

[0034] Compound 5: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0035]

[0036] Compound 6: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0037]

[0038] Compound 7: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1,2,4-triazolyl)phenyl)-1,3,4-thiadiazole-2-amine

[0039]

[0040] Compound 8: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazole-2-amine

[0041]

[0042] Compound 9: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0043]

[0044] Compound 10: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0045]

[0046] Compound 11: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0047]

[0048] Compound 12: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0049]

[0050] Compound 13: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(3-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0051]

[0052] Compound 14: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0053]

[0054] Compound 15: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazole-2-amine

[0055]

[0056] Compound 16: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(3-hydroxyphenyl)-1,3,4-thiadiazole-2-amine

[0057]

[0058] Compound 17: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0059]

[0060] Compound 18: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1-pyrrolidinyl)phenyl)-1,3,4-thiadiazole-2-amine

[0061]

[0062] Compound 19: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0063]

[0064] Compound 20: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylphenyl)-1,3,4-thiadiazole-2-amine

[0065]

[0066] Compound 21: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-(3-trifluoromethylphenyl)oxyphenyl)-1,3,4-thiadiazole-2-amine

[0067]

[0068] Compound 22: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylthiophenyl)-1,3,4-thiadiazole-2-amine

[0069]

[0070] Compound 23: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(phenyl)phenyl)-1,3,4-thiadiazole-2-amine

[0071]

[0072] Compound 24: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4-dichlorophenyl)-1,3,4-thiadiazole-2-amine

[0073]

[0074] Compound 25: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(morpholinyl)phenyl)-1,3,4-thiadiazole-2-amine

[0075]

[0076] Compound 26: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(methoxy)phenyl)-1,3,4-thiadiazole-2-amine

[0077]

[0078] Compound 27: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0079]

[0080] Compound 28: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethyloxyphenyl)-1,3,4-thiadiazole-2-amine

[0081]

[0082] Compound 29: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0083]

[0084] Compound 30: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0085]

[0086] Compound 31: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0087]

[0088] Compound 32: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazole-2-amine

[0089]

[0090] Compound 33: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0091]

[0092] Compound 34: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0093]

[0094] Compound 35: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0095]

[0096] Compound 36: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-hydroxyphenyl)-1,3,4-thiadiazole-2-amine

[0097]

[0098] Compound 37: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-chlorophenyl)-1,3,4-thiadiazole-2-amine

[0099]

[0100] Compound 38: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-chlorophenyl)-1,3,4-thiadiazole-2-amine

[0101]

[0102] Compound 39: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-chlorophenyl)-1,3,4-thiadiazole-2-amine

[0103]

[0104] Compound 40: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-bromophenyl)-1,3,4-thiadiazole-2-amine

[0105]

[0106] Compound 41: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-bromophenyl)-1,3,4-thiadiazole-2-amine

[0107]

[0108] Compound 42: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0109]

[0110] Compound 43: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazole-2-amine

[0111]

[0112] Compound 44: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0113]

[0114] Compound 45: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0115]

[0116] Compound 46: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-propoxyphenyl)-1,3,4-thiadiazole-2-amine

[0117]

[0118] Compound 47: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-ethynylphenyl)-1,3,4-thiadiazole-2-amine

[0119]

[0120] Compound 48: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3,4-ethylenedioxyphenyl)-1,3,4-thiadiazole-2-amine

[0121]

[0122] Compound 49: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-formamidophenyl)-1,3,4-thiadiazole-2-amine

[0123]

[0124] Compound 50: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-iodophenyl)-1,3,4-thiadiazole-2-amine

[0125]

[0126] Compound 51: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-iodophenyl)-1,3,4-thiadiazole-2-amine

[0127]

[0128] Compound 52: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-iodophenyl)-1,3,4-thiadiazole-2-amine

[0129]

[0130] Compound 53: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-hydroxyphenyl)-1,3,4-thiadiazole-2-amine

[0131]

[0132] Compound 54: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-ethoxyacylphenyl)-1,3,4-thiadiazole-2-amine

[0133]

[0134] Compound 55: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-ethoxyacylphenyl)-1,3,4-thiadiazole-2-amine

[0135]

[0136] Compound 56: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-methanesulfonylaminophenyl)-1,3,4-thiadiazole-2-amine

[0137]

[0138] Compound 57: N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0139]

[0140] Compound 58: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0141]

[0142] Compound 59: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-isopropylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0143]

[0144] Compound 60: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0145]

[0146] Compound 61: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0147]

[0148] Compound 62: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0149]

[0150] Compound 63: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0151]

[0152] Compound 64: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0153]

[0154] Compound 65: N-(1-isopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0155]

[0156] Compound 66: N-(1-pentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0157]

[0158] Compound 67: N-(1-cyclopentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0159]

[0160] Compound 68: N-(1-cyclopropyl-2-phenyl-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0161]

[0162] Compound 69: N-(1-cyclopropyl-2-(4-chlorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0163]

[0164] Compound 70: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0165]

[0166] Compound 71: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0167]

[0168] Compound 72: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0169]

[0170] Compound 73: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0171]

[0172] Compound 74: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0173]

[0174] Compound 75: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0175]

[0176] Compound 76: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-fluoro-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0177]

[0178] Compound 77: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0179]

[0180] Compound 78: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0181]

[0182] Compound 79: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0183]

[0184] Compound 80: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0185]

[0186] Compound 81: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0187]

[0188] Compound 82: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0189]

[0190] On the other hand, the present invention also relates to a complete preparation route and preparation method of the compound of the general formula I:

[0191]

[0192] In the above formula, R1-R4 are as defined above.

[0193] The synthesis method of the present invention is described in detail below.

[0194] In the above reaction, compound 1 is commercially available. Compound 4, a polysubstituted 5-aminobenzimidazole active core derivative, can be obtained from compound 1 through monosubstitution (a), disubstitution (b), reduction (c), and ring closure (d) according to the literature (J. Comb. Chem. 2004, 6, 811-821).

[0195] In step (e), the reagents used are thiocarbonyldiimidazole and triethylamine, preferably in a molar ratio of 1:2.2 to compound 4. The solvent is an aprotic solvent such as dichloromethane. The temperature is 0°C to RT, and the reaction time is 1 hour. In step (f), hydrazine hydrate is then added to the reaction solution. The molar ratio of hydrazine to compound 4 is 1:2. The temperature is 80°C, and the reaction time is 1 to 24 hours, preferably 3 hours, to obtain compound 5.

[0196] The reagents used in step (g) are aldehydes with corresponding R4 substituents, such as triazole benzaldehyde, pyridine carboxaldehyde, 2-fluorobenzaldehyde, 3-fluorobenzaldehyde, 4-fluorobenzaldehyde, 2-bromobenzaldehyde, 3,4,5-trimethoxybenzaldehyde, 4-acetamidobenzaldehyde, 3-fluorobenzaldehyde, and 3-trifluoromethylbenzaldehyde. The solvent is a protic solvent such as ethanol. The catalyst is 5% to 8% CH3COOH. The temperature is 80°C and the reaction time is 1 hour. The reagent used in step (h) is ferric chloride hexahydrate in a molar ratio of 1:2.5 to compound 5. The temperature is 80°C and the reaction time is 12 to 24 hours to obtain I.

[0197] The above reaction is advantageous for the synthesis of compound libraries using, for example, combinatorial chemistry methods due to its relatively mild conditions, short reaction time, stable yield, and strong versatility. Such methods for synthesizing compound libraries using combinatorial chemistry methods also fall within the scope of the present invention.

[0198] Those skilled in the art may modify the above steps to improve yield. They may determine the synthetic route based on basic knowledge in the art, such as selecting reactants, solvents, and temperatures. Such modifications or variations are within the scope of the present invention. Yields may also be improved by using various conventional protecting groups to avoid side reactions. These conventional protection methods can be found, for example, in Protecting Groups in Organic Synthesis (T. Greene, the Fourth Edition, John Wiley & Sons, Inc.).

[0199] The invention relates to the use of the general formula of the compound in the preparation of drugs for preventing, alleviating and / or treating symptoms caused by cancer, inflammation, and autoimmune diseases. As long as these benzimidazole compounds and their derivatives are used for cancer and its complications, they all fall within the scope of protection of the compounds of the present invention.

[0200] The cancers mentioned particularly refer to breast cancer, prostate cancer, ovarian cancer, liver cancer, stomach cancer, lung cancer, colon cancer, pancreatic cancer, esophageal cancer, leukemia, human brain glioma, and lymphoma.

[0201] In the present invention, the prevention, alleviation and / or treatment of cancer or symptoms is selected from the group consisting of inhibiting the activity of STAT3, inhibiting the expression of genes related to STAT3, and inhibiting the proliferation and metastasis of breast cancer, prostate cancer, ovarian cancer, liver cancer, gastric cancer, lung cancer, colon cancer, pancreatic cancer, esophageal cancer, leukemia, human brain glioma and lymphoma.

[0202] The present invention is achieved through the following technical solutions: a series of benzimidazol-2-amino-1,3,4-thiadiazole derivatives are prepared by artificial synthesis; the inhibitory activity of the compounds on the IL-6-STAT3 pathway is evaluated using a HEK-BLUE-IL-6 cell model; the interaction between the compounds and the STAT3 protein is evaluated by circular dichroism experiments; the inhibitory activity of the compounds on MDA-MB-231 and MDA-MB-468 tumor cells is evaluated by MTT experiments; and the selective inhibitory activity of the compounds on STAT3 is evaluated by Western Blotting experiments.

[0203] The benzimidazolo-2-amino-1,3,4-thiadiazole compounds of the present invention, generally represented by Formula I, have a pharmaceutical effect in preventing, alleviating, and / or treating cancer when formulated into any dosage form. Any pharmaceutical agent containing a benzimidazolo-2-amino-1,3,4-thiadiazole compound of Formula I, or formulated solely with a benzimidazolo-2-amino-1,3,4-thiadiazole compound of Formula I, also falls within the scope of protection of the present invention, provided that the agent's effect of treating cancer and its complications is noted or indicated on its packaging, instructions, or other promotional materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0204] Figure 1 .Circular dichroism curves of the interaction between STAT3 and the example compounds

[0205] Figure 2 Figure 1. Cellular thermal migration assay of the interaction between STAT3 and the example compounds

[0206] Figure 3 Immunoblotting was used to detect the inhibition of p-STAT3 in MDA-MB-231 cells by the example compounds and positive control drugs.

[0207] Figure 4 Immunoblotting was used to detect the inhibition of p-STAT3 in MDA-MB-231 cells by the example compounds.

[0208] Figure 5 Immunoblotting was used to detect the inhibition of p-STAT3 in MDA-MB-468 cells by the example compounds. DETAILED DESCRIPTION

[0209] Example

[0210] The present invention will be further described below with reference to the following examples, but the present invention is not limited to these examples.

[0211] Chemistry Experiment Section

[0212] Melting points were determined using an RY-2 melting point apparatus from Tianjin Analytical Instrument Factory; temperatures are uncorrected. Mass spectra were measured using a ThermoFinnigan LCQ-Advantage mass spectrometer. H and C NMR spectra were measured using a Varian Mercury 300 MHz, 400 MHz, 500 MHz, or 700 MHz NMR spectra, using CDCl₃ or DMSO-d₆ as solvents. Mass spectra were measured using a Thermo Exactive-Orbitrap mass spectrometer.

[0213] Example 1 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0214] Step 1: 1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-aminobenzimidazole

[0215]

[0216] Compound 1 (1 mmol, 204.1 mg) was dissolved in 10 mL of tetrahydrofuran, N,N-diisopropylethylamine (DIPEA, 1.2 mmol) was added, and then a solution of cyclopropylamine (1.1 mmol) in 2 mL of tetrahydrofuran was slowly added dropwise and allowed to react at room temperature. After the reaction was complete, DIPEA (1.2 mmol) was added, followed by the slow addition of N-ethylpiperazine (1 mmol) and the reaction was allowed to react at room temperature. After the reaction was complete, a saturated ammonium chloride solution was added, and the mixture was extracted with dichloromethane and concentrated to obtain a disubstituted product. The disubstituted intermediate (1 mmol) was dissolved in a mixed solution of 5 mL of ethanol and 5 mL of tetrahydrofuran, and 1.0 eq of 10% Pd-C and 7.5 eq of HCOONH4 were added with stirring, and the mixture was stirred at room temperature for 1 hour. The reaction solution was quickly filtered into a solution of 1.2 mmol of p-fluorobenzaldehyde in 5 mL of tetrahydrofuran, and 1 mL of acetic acid was added, and the mixture was stirred at room temperature. HPLC-MS detection was performed until the reaction was complete, and the solvent was evaporated under reduced pressure. The pH was adjusted to neutral with saturated sodium bicarbonate solution. Extraction was performed with dichloromethane, drying was performed with anhydrous sodium sulfate, and the organic phase was concentrated. Purification was performed by column chromatography with ethyl acetate-petroleum ether as the eluent to obtain a brown solid. Melting point: 200-204°C. ESI-MS (m / z): calcd. for C 22 H 27 FN5[M+H] + ,380.23,found380.23.

[0217] Step 2: 1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-thiosemicarbazide benzimidazole

[0218]

[0219] The compound 1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-aminobenzimidazole (1 mmol) was dissolved in 5 mL of anhydrous dichloromethane and 2.2 mmol of triethylamine was added with stirring. 1 mmol of thiocarbonyldiimidazole was slowly added dropwise under an ice bath and allowed to react at room temperature for 1 hour. 1 mmol of hydrazine hydrate was then added to the reaction solution and heated under reflux for 1 hour. HPLC-MS detection was performed until the reaction was complete, and the reaction solution was concentrated under reduced pressure. Column chromatography was performed and eluted with ethyl acetate-petroleum ether to obtain a white solid. Melting point: 126-128°C. ESI-MS (m / z): calcd.for C 23 H 29 FN7S[M+H] + ,454.22,found 454.23.

[0220] Step 3: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0221]

[0222] Dissolve 1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-thiosemicarbazide benzimidazole (227 mg, 0.5 mmol) in 25 mL of ethanol, add benzaldehyde (0.5 mmol) and acetic acid (1 mL), and heat under reflux for 1 hour. Add FeCl₃·6H₂O to the reaction solution and react at 80°C for 5 hours. Neutralize with saturated NaHCO₃ solution, extract with dichloromethane, and purify the extract by column chromatography to obtain a yellow solid. Melting point: 264-266°C. ESI-HRMS (m / z): calcd. for C 30 H 31 FN7S[M+H] + ,540.2346,found540.2337. 1 H NMR (400MHz, DMSO-d6) δ9.58(s,1H),8.28(s,1H),8.07(dd,J=8.7,5.6Hz,2H),7.90-7.81(m,2H),7.57-7.47(m,3H),7.45-7.37(m,3H),3. 81(tt,J=7.0,3.7Hz,1H),3.35(s,4H),3.09(s,4H),2.91(s,2H),1.17(d,J=6.8Hz,3H),1.14(t,J=3.6Hz,2H),0.66(q,J=5.2,4.1Hz,2H).

[0223] Example 2 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0224]

[0225] The same operation as in Example 1 was performed, except that p-trifluoromethylbenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 193-195°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F4N7S[M+H] + ,608.2220,found 608.2212. 1 H NMR (400MHz, CDCl3) δ8.87(s,1H),7.96(q,J=8.6,8.2Hz,5H),7.71(d,J=8.4Hz,2H),7.50(s,1H),7.21(t,J=8.5Hz,2H),3.80-3.65(m, 1H),3.52(q,J=5.3,4.3Hz,1H),3.15(s,4H),3.04-2.79(m,3H),2.71(s,2H),1.25(d,J=5.9Hz,3H),1.15(d,J=6.7Hz,2H),0.75(s,2H).

[0226] Example 3 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0227]

[0228] The same operation as in Example 1 was performed, except that 4-fluorobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 192-194°C. ESI-HRMS (m / z): calcd. for C 30 H 30 F2N7S[M+H] + ,558.2251,found558.2249. 1HNMR(500MHz,DMSO-d6)δ9.47(s,1H),8.12(s,1H),8.03-7.97(m,2H),7.86-7.80(m,2H),7.38-7.27(m,5H),3.73(tt,J=7.1,3.8 Hz,1H),2.93(t,J=4.8Hz,4H),2.55(s,4H),2.37(p,J=7.4,6.3Hz,2H),1.12-1.06(m,2H),1.02-0.97(m,3H),0.63-0.57(m,2H).

[0229] Example 4 N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0230]

[0231] The procedure was the same as in Example 1, except that in step 1, a monosubstituted cyclopropylamine was used, and in step 3, p-nitrobenzaldehyde was used instead of benzaldehyde to obtain N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange-yellow solid. Melting point: 241-243°C. ESI-HRMS (m / z): calcd. for C 24 H 17 F2N6O2S[M+H] + ,491.1102,found 491.1095. 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.53(d,J=7.6Hz,1H),8.33(d,J=8.3Hz,2H),8.09(dd,J=27.7,7. 6Hz, 4H), 7.63 (d, J = 10.8Hz, 1H), 7.41 (t, J = 8.6Hz, 2H), 3.80 (s, 1H), 1.12 (d, J = 6.9Hz, 2H), 0.66 (s, 2H).

[0232] Example 5 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0233]

[0234] The same operation as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 161-163°C. ESI-HRMS (m / z): calcd. for C 30 H 30 F2N7S[M+H] + ,558.2251,found558.2244. 1 H NMR (400MHz, CDCl3) δ8.90 (s, 1H), 8.29 (td, J = 7.6, 1.8Hz, 1H), 8.04-7.87 (m, 3H) ,7.49(s,1H),7.42(tdd,J=7.5,5.2,1.9Hz,1H),7.28(d,J=7.7Hz,1H),7.24-7.16 (m,3H),3.52(tt,J=7.1,3.8Hz,1H),3.25(s,4H),3.07(s,4H),2.85(q,J=7.7,7.2 Hz, 2H), 1.32 (t, J = 7.2Hz, 3H), 1.14 (p, J = 6.6Hz, 2H), 0.74 (dd, J = 4.4, 2.4Hz, 2H).

[0235] Example 6 N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0236]

[0237] The procedure was the same as in Example 1, except that in step 1, a monosubstituted cyclopropylamine was used, and in step 3, p-cyanobenzaldehyde was used instead of benzaldehyde to obtain N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 239-241°C. ESI-HRMS (m / z): calcd. for C 25 H 17 F2N6S[M+H] + ,471.1125,found471.1194. 1H NMR (400MHz, DMSO-d6) δ10.46 (s, 1H), 8.55 (d, J = 7.0Hz, 1H), 8.02 (dd, J = 31.7, 8.3Hz, 6H), 7. 64(d,J=10.9Hz,1H),7.42(t,J=8.9Hz,2H),3.81(s,1H),1.13(d,J=6.9Hz,2H),0.68(s,2H).

[0238] Example 7 N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1,2,4-triazolyl)phenyl)-1,3,4-thiadiazole-2-amine

[0239]

[0240] The procedure was the same as in Example 1, except that in step 1, a monosubstituted cyclopropylamine was used, and in step 3, p-1,2,4-triazole benzaldehyde was used instead of p-fluorobenzaldehyde to obtain N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1,2,4-triazolyl)phenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 260-262°C. ESI-HRMS (m / z): calcd. for C 26 H 19 F2N8S[M+H] + ,513.1421,found 513.1415. 1 H NMR (400MHz, DMSO-d6) δ10.35(s,1H),9.43(s,1H),8.56(d,J=7.3Hz,1H),8.31(s,1H),8.14-7.99(m ,6H),7.64(d,J=10.8Hz,1H),7.42(t,J=8.7Hz,2H),3.82(s,1H),1.14(d,J=6.7Hz,2H),0.68(s,2H).

[0241] Example 8 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazole-2-amine

[0242]

[0243] The same operation as in Example 1 was performed, except that p-dimethylaminobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazol-2-amine as a brown solid. Melting point: 181-182°C. ESI-HRMS (m / z): calcd. for C 32 H 36 FN8S[M+H] + ,583.2768,found 583.2764. 1 H NMR (400MHz, CDCl3) δ7.99(dd,J=11.6,5.1Hz,4H),7.76(d,J=8.4Hz,1H),7.48(d,J=25.4Hz,2H),7.30(s,2H),6.48(d, J=8.5Hz,2H),3.54(s,1H),3.24(s,3H),3.06(s,2H),2.94(s,6H),2.66(s,5H),1.22(s,3H),0.91(s,2H),0.73(m,2H).

[0244] Example 9 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0245]

[0246] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used in place of p-fluorobenzaldehyde in step 1 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine as a brown solid. Melting point: 169-170°C. ESI-HRMS (m / z): calcd. for C 30 H 31 N7FS[M+H] + ,540.2346,found 540.2340. 1H NMR (400MHz, CDCl3) δ7.96 (s, 1H), 7.87-7.79 (m, 2H), 7.72 (td, J = 7.4, 1.8Hz ,1H),7.59-7.40(m,6H),7.32(t,J=7.5Hz,1H),7.25-7.21(m,1H),3.71(t,J= 6.9Hz,1H),3.51(m,J=7.0,3.6,3.2Hz,4H),3.41-3.15(m,4H),3.10(d,J=6.7 Hz,2H),1.48(q,J=5.4,3.7Hz,3H),1.02(t,J=6.6Hz,2H),0.69-0.60(m,2H).

[0247] Example 10 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0248]

[0249] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and p-trifluoromethylbenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 239-241°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F4N7S[M+H] + ,608.2220,found 608.2214. 1 H NMR (400MHz, CDCl3) δ8.77(s,1H),8.03-7.93(m,3H),7.75-7.69(m,3H),7.59-7.49(m,2H),7.32(td,J=7.5,1.1Hz,1H),7.25-7.21(m,1H),3.72( d,J=7.1Hz,1H),3.53(tt,J=6.4,3.2Hz,1H),3.21(s,4H),2.97(s,3H),2 .79(s,2H),1.30(s,3H),1.05-0.97(m,2H),0.66(dd,J=6.6,3.9Hz,2H).

[0250] Example 11 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0251]

[0252] The same operation as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1 and p-fluorobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 240-242°C. ESI-HRMS (m / z): calcd. for C 30 H 30 F2N7S[M+H] + ,558.2251,found 558.2247. 1 H NMR (400MHz, DMSO-d6) δ9.62(s,1H),8.28(s,1H),7.94-7.85(m,2H),7.69(dtd,J=35.5,7.5,1.8Hz,2H),7.49-7.31(m,5H) ,3.55(s,1H),3.35(s,4H),3.06(s,4H),2.76(s,2H),1.12(s,3H),0.97(dd,J=7.4,5.4Hz,2H),0.60(p,J=5.3,4.8Hz,2H).

[0253] Example 12 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0254]

[0255] The same procedure as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1 and o-fluorobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 204-206°C. ESI-HRMS (m / z): calcd. for C 30 H 30 F2N7S[M+H] + ,558.2251,found 558.2245. 1H NMR(400MHz, CDCl3)δ8.78(s,1H),8.33(td,J=7.6,1.8Hz,1H),8.03(s,1H),7.73(td,J=7.4,1.8Hz, 1H),7.57-7.49(m,2H),7.42(dddd,J=8.4,7.2,5.3,1.8Hz,1H),7.33(dd,J=7.6,1.1Hz,1H),7.30-7 .27(m,1H),7.23(d,J=1.2Hz,1H),7.21-7.17(m,1H),3.72(q,J=7.0Hz,1H),3.59-3.48(m,1H),3.12 (s,4H),2.84(s,3H),2.66(s,2H),1.24-1.20(m,3H),0.99(td,J=7.2,5.5Hz,2H),0.70-0.62(m,2H).

[0256] Example 13 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(3-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0257]

[0258] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and p-trifluoromethylbenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(3-trifluoromethylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 278-280°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F4N7S[M+H] + ,608.2141,found 608.2217. 1H NMR (400MHz, CDCl3) δ8.73 (s, 1H), 8.13 (d, J = 2.1Hz, 1H), 7.98 (d, J = 9.1Hz, 2H), 7.7 6-7.65(m,2H),7.63-7.50(m,3H),7.32(td,J=7.5,1.1Hz,1H),7.26-7.21(m,1H),3. 78(d,J=11.0Hz,1H),3.72(q,J=7.0Hz,1H),3.51(tt,J=6.6,3.2Hz,2H),3.28(s,5H ),3.11(s,2H),1.50(t,J=7.2Hz,3H),1.08-0.99(m,2H),0.65(p,J=5.7,5.0Hz,2H).

[0259] Example 14 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0260]

[0261] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and p-trifluoromethoxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethoxyphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 264-266°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F4N7OS[M+H] + ,624.2169,found624.2165. 1 H NMR(400MHz, CDCl3)δ8.68(s,1H),7.96(s,1H),7.93-7.86(m,2H),7.72( td,J=7.4,1.8Hz,1H),7.58-7.49(m,2H),7.36-7.28(m,3H),7.25-7.20(m ,1H),3.53(m,J=7.1,4.9,2.2Hz,1H),3.25(s,4H),3.04(s,4H),2.85(s,2 H),1.35(m,J=8.2,7.7,4.1Hz,3H),1.06-0.94(m,2H),0.70-0.60(m,2H).

[0262] Example 15 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazole-2-amine

[0263]

[0264] The same procedure as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and 4-dimethylaminobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 182-185°C. ESI-HRMS (m / z): calcd. for C 32 H 36 FN8S[M+H] + ,583.2768,found583.2769. 1 H NMR (400MHz, CDCl3) δ8.73 (s, 1H), 7.95 (s, 1H), 7.70 (td, J = 7.4, 1.8Hz, 1H), 7.66-7.60 ( m,2H),7.52(dddd,J=8.4,7.2,5.2,1.8Hz,1H),7.47(s,1H),7.30(td,J=7.6,1.1Hz,1H), 7.22(ddd,J=9.6,8.4,1.1Hz,1H),6.72-6.65(m,2H),3.70(q,J=7.0Hz,1H),3.52-3.46(m ,2H),3.19-3.06(m,8H),3.01(s,6H),1.38(s,3H),1.04-0.96(m,2H),0.66-0.58(m,2H).

[0265] Example 16 N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(3-hydroxyphenyl)-1,3,4-thiadiazole-2-amine

[0266]

[0267] The same procedure as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and 4-acetoxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-(acetoxy)phenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 167-169°C. ESI-HRMS (m / z): calcd. for C 32 H 33 FN7O2S[M+H] + ,556.2217,found556.2288. 1 H NMR (400MHz, CDCl3) δ8.85 (s, 1H), 7.88 (d, J = 12.6Hz, 2H), 7.77 (t, J = 7.4Hz, 1H), 7 .54(d,J=12.6Hz,2H),7.41-7.32(m,2H),7.24(dd,J=19.3,8.4Hz,2H),7.08-7.00( m,1H),3.64-3.50(m,1H),3.10(t,J=4.6Hz,4H),2.82(s,3H),2.64(q,J=7.1Hz,2H ), 2.47 (s, 1H), 1.21 (t, J = 7.2Hz, 3H), 1.01 (t, J = 6.7Hz, 2H), 0.70 (d, J = 3.3Hz, 2H).

[0268] Example 17 N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0269]

[0270] The procedure was the same as in Example 1, except that the monosubstituted cyclopropylamine was used in step 1 and 2-nitrobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange solid. Melting point: 256-258°C. ESI-HRMS (m / z): calcd. for C 24 H 17 F2N6O2S[M+H] + ,491.1102,found491.1100. 1H NMR (400MHz, DMSO-d6) δ10.41 (s, 1H), 8.55 (d, J = 7.4Hz, 1H), 8.13-8.03 (m, 3H), 7.92-7.75 (m, 3H), 7.64 (d, J=10.7Hz,1H),7.42(t,J=8.6Hz,2H),3.83(d,J=10.4Hz,1H),1.13(d,J=6.8Hz,2H),0.68(d,J=3.8Hz,2H).

[0271] Example 18 N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1-pyrrolidinyl)phenyl)-1,3,4-thiadiazole-2-amine

[0272]

[0273] The procedure was the same as in Example 1, except that in step 1, a monosubstituted cyclopropylamine was used, and in step 3, 1-pyrrolidinylbenzaldehyde was used instead of benzaldehyde to obtain N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1-pyrrolidinyl)phenyl)-1,3,4-thiadiazol-2-amine as a blue-purple solid. Melting point: 256-258°C. ESI-HRMS (m / z): calcd. for C 28 H 25 F2N6S[M+H] + ,515.1829,found 515.1827. 1 H NMR (400MHz, DMSO-d6) δ13.98(s,1H),8.09(dd,J=8.5,5.4Hz,2H),7.92(d,J=6.7Hz,1H),7.72(d,J=9.7Hz,1H),7.44(t,J=8.7Hz,2H),7.19(d,J=8.5Hz ,2H),6.44(d,J=8.6Hz,2H),3.84(dd,J=7.7,3.7Hz,1H),3.35(s,2H),1.99 -1.83(m,4H),1.28(d,J=13.4Hz,2H),1.20-1.09(m,2H),0.79-0.59(m,2H).

[0274] Example 19 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0275]

[0276] The same operation as in Example 1 was performed, except that benzaldehyde was replaced with m-cyanobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-cyanophenyl)-1,3,4-thiadiazole-2-amine as a brown solid. Melting point: 193-195°C. ESI-HRMS (m / z): calcd. for C 31 H 30 FN8S[M+H] + ,565.2298,found565.2291. 1 H NMR (400MHz, DMSO-d6) δ9.70 (s, 1H), 8.27 (d, J = 1.9Hz, 1H), 8.21-8.13 (m, 2H ),8.10-8.02(m,2H),7.94(d,J=7.6Hz,1H),7.72(t,J=7.9Hz,1H),7.46-7.3 7(m,3H),3.80(tt,J=7.1,3.9Hz,1H),3.00(t,J=4.8Hz,4H),2.61(s,4H),2. 47-2.38(m,2H),1.20-1.13(m,2H),1.08-1.03(m,3H),0.66(p,J=4.8Hz,2H).

[0277] Example 20 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylphenyl)-1,3,4-thiadiazole-2-amine

[0278]

[0279] The same operation as in Example 1 was performed, except that p-methylbenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 248-250°C. ESI-HRMS (m / z): calcd. for C 31 H 33 FN7S[M+H] + ,554.2502,found554.2495. 1H NMR (400MHz, CDCl3) δ8.07-7.86(m,4H),7.47(s,1H),7.24-7.20(m,3H),7.03(d,J=8.0Hz,3H),3.69-3.59(m,1H),3. 54-3.46(m,2H),3.32(s,4H),3.03(s,3H),2.28(s,4H),1.21-1.16(m,2H),0.85(dd,J=14.7,6.9Hz,3H),0.65(s,2H).

[0280] Example 21 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-(3-trifluoromethylphenyl)oxyphenyl)-1,3,4-thiadiazole-2-amine

[0281]

[0282] The operation was the same as in Example 1, except that 3-trifluoromethylphenyloxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-(3-trifluoromethylphenyl)oxyphenyl)-1,3,4-thiadiazole-2-amine as a yellow oily liquid. ESI-HRMS (m / z): calcd. for C 37 H 34 F4N7OS[M+H] + ,700.2482,found 700.2479. 1 H NMR (400MHz, CDCl3) δ8.00-7.83(m,3H),7.68(dt,J=7.6,3.8Hz,1H),7.58(t,J=5.5H z,1H),7.54-7.32(m,5H),7.23-7.10(m,4H),7.07(dd,J=8.2,2.5Hz,1H),4.21(d,J=5 .8Hz,1H),3.50(tt,J=7.3,3.9Hz,1H),3.06(t,J=4.9Hz,4H),2.76(s,3H),2.58(q,J =7.1Hz,2H),1.17(t,J=7.2Hz,3H),1.11(d,J=6.8Hz,2H),0.72(q,J=6.2,4.5Hz,2H).

[0283] Example 22 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylthiophenyl)-1,3,4-thiadiazole-2-amine

[0284]

[0285] The same operation as in Example 1 was performed, except that 4-methylthiobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylthiophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 153-155°C. ESI-HRMS (m / z): calcd. for C 31 H 33 FN7S2[M+H] + ,586.2223,found586.2220. 1 H NMR (400MHz, CDCl3) δ8.78(s,1H),8.05-7.94(m,4H),7.81(d,J=8.2Hz,2H),7.52(s,1H),7.38(s,1H),7.22(s,1H),7.06(d,J=8.3Hz,1H),3.5 4(d,J=8.4Hz,1H),3.17(s,4H),2.91(s,2H),2.72(s,2H),2.56(s,3H), 2.44(s,2H),1.17(d,J=6.8Hz,3H),0.90(d,J=9.4Hz,2H),0.78(s,2H).

[0286] Example 23 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(phenyl)phenyl)-1,3,4-thiadiazole-2-amine

[0287]

[0288] The same operation as in Example 1 was performed, except that 4-benzaldehydebenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(phenyl)phenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 140-141°C. ESI-HRMS (m / z): calcd. for C 36 H 35 FN7S[M+H] + ,616.2659,found616.2659. 1H NMR (400MHz, CDCl3) δ8.03-7.92(m,5H),7.78-7.71(m,2H),7.70-7.65(m,2H),7.57-7.48(m,3H),7.47-7.40(m,1H),7.29-7.18(m,3H),3. 57(dq,J=6.9,3.4Hz,1H),3.50(dq,J=6.8,3.4Hz,1H),3.36(s,6H),3.02(s,3H),0.93-0.91(m,3H),0.81-0.76(m,2H),0.75-0.70(m,1H).

[0289] Example 24 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4-dichlorophenyl)-1,3,4-thiadiazole-2-amine

[0290]

[0291] The same operation as in Example 1 was performed, except that 3,4-dichlorobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4-dichlorophenyl)-1,3,4-thiadiazol-2-amine as a brown solid. Melting point: 122-124°C. ESI-HRMS (m / z): calcd. for C 30 H 29 Cl2FN7S[M+H] + ,608.1566,found608.1566. 1 H NMR (400MHz, CDCl3) δ8.80(s,1H),8.03-7.93(m,3H),7.78(d,J=1.8Hz,2H),7.55(s,1H),7.44(t,J=1.8Hz,1H),7.28-7.21(m,2H),3.5 6(tq,J=7.6,3.7Hz,1H),3.27(s,4H),3.06(s,4H),2.87(s,2H),1.23-1.18(m,3H),0.91(d,J=2.6Hz,2H),0.79(dq,J=4.7,1.7Hz,2H).

[0292] Example 25 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(morpholinyl)phenyl)-1,3,4-thiadiazole-2-amine

[0293]

[0294] The same procedure as in Example 1 was performed, except that 4-morpholinylbenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(morpholinyl)phenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 149-151°C. ESI-HRMS (m / z): calcd. for C 34 H 38 FN8OS[M+H] + ,625.2873,found625.2873. 1 H NMR(400MHz, CDCl3)δ8.01-7.86(m,4H),7.80-7.72(m,1H),7.49(s,1H),7.23-7.17(m ,2H),6.97-6.91(m,1H),6.88(dd,J=8.9,5.3Hz,1H),6.81-6.66(m,1H),3.93-3.84(m ,4H),3.83-3.78(m,2H),3.52(m,1H),3.41-3.30(m,2H),3.30-3.22(m,4H),3.18-3.0 5(m,4H),2.92(s,2H),1.15(t,J=6.7Hz,3H),0.88(d,J=6.3Hz,2H),0.76-0.71(m,2H).

[0295] Example 26 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(methoxyl)phenyl)-1,3,4-thiadiazole-2-amine

[0296]

[0297] The same operation as in Example 1 was performed, except that p-methoxyacylbenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(methoxyacyl)phenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 149-151°C. ESI-HRMS (m / z): calcd. for C 32 H 33 FN7O2S[M+H] + ,598.2400,found 598.2400. 1H NMR (400MHz, CDCl3) δ8.74(s,1H),8.16-8.08(m,2H),8.00-7.86(m,5H),7.51(s,1H),7.25-7.18(m,2H),3.95(s,3H),3. 53(tt,J=7.0,3.7Hz,1H),3.23(s,4H),3.01(s,4H),2.82(s,2H),1.32(s,3H),1.16(t,J=6.5Hz,2H),0.78-0.71(m,2H).

[0298] Example 27 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0299]

[0300] The same procedures as in Example 1 were performed, except that N-methylpiperazine was used instead of N-ethylpiperazine in step 1, and p-fluorobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 156-158°C. ESI-HRMS (m / z): calcd. for C 29 H 28 F2N7S[M+H] + ,544.2095,found 544.2095. 1 H NMR(400MHz, CDCl3)δ8.73(s,1H),8.06-7.94(m,3H),7.89(dd,J=8.4,5.4Hz,2H),7.50(s,1H),7.22(m,J=23.6,8.4Hz,4 H), 3.56 (d, J = 6.7Hz, 1H), 3.12 (t, J = 4.8Hz, 4H), 2.82 (s, 4H), 2.52 (s, 3H), 1.19 (t, J = 6.8Hz, 2H), 0.79 (d, J = 4.3Hz, 2H).

[0301] Example 28 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethyloxyphenyl)-1,3,4-thiadiazole-2-amine

[0302]

[0303] The same procedures as in Example 1 were performed, except that N-methylpiperazine was used instead of N-ethylpiperazine in step 1, and p-trifluoromethoxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethyloxyphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 184-185°C. ESI-HRMS (m / z): calcd. for C 30 H 28 F4N7OS[M+H] + ,610.2012,found 610.2011. 1 H NMR(400MHz,Chloroform-d)δ8.02-7.89(m,5H),7.49(d,J=1.6Hz,1H),7.34(d,J=8.3Hz,2H),7.25(m,J=8.7,1.6H z,2H),3.56(s,1H),3.13(s,4H),2.87(s,4H),2.53(d,J=2.4Hz,3H),1.19(d,J=6.8Hz,2H),0.79(d,J=4.1Hz,2H).

[0304] Example 29 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0305]

[0306] The same operation as in Example 1 was performed, except that p-nitrobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 165-167°C. ESI-HRMS (m / z): calcd. for C 30 H 30 FN8O2S[M+H] + ,585.2196,found585.2196. 1H NMR (400MHz, CDCl3) δ8.95(s,1H),8.36(d,J=8.5Hz,2H),8.08(d,J=8.7Hz,2H),8.04-7.95(m,3H),7.54(s,1H),7.30(s,2H ),3.57(s,1H),3.14(s,4H),2.86(s,3H),2.68(s,2H),1.28-1.23(m,3H),1.19(d,J=6.6Hz,2H),0.92(s,1H),0.80(s,2H).

[0307] Example 30 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0308]

[0309] The same procedure as in Example 1 was performed, except that benzaldehyde was replaced with m-nitrobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 134-135°C. ESI-HRMS (m / z): calcd. for C 30 H 30 FN8O2S[M+H] + ,585.2196,found585.2196. 1 H NMR (400MHz, CDCl3) δ8.82 (s, 1H), 8.74 (s, 1H), 8.31 (d, J = 8.3Hz, 1H), 8.23 ​​( d,J=7.8Hz,1H),7.99(d,J=10.6Hz,3H),7.69(t,J=8.0Hz,1H),7.55(s,1H), 7.26(t,J=8.5Hz,2H),3.57(s,1H),3.26(s,4H),3.04(s,3H),2.85(s,2H),1 .35(s,2H),1.20(d,J=6.9Hz,2H),0.92(d,J=7.1Hz,2H),0.81-0.76(m,2H).

[0310] Example 31 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0311]

[0312] The same operation as in Example 1 was performed, except that o-nitrobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 134-135°C. ESI-HRMS (m / z): calcd. for C 30 H 30 FN8O2S[M+H] + ,585.2196,found585.2196. 1 H NMR (400MHz, CDCl3) δ8.88(s,1H),7.97(dt,J=8.4,4.0Hz,4H),7.84(d,J=7.7Hz,1 H),7.74(t,J=7.6Hz,1H),7.65(t,J=7.8Hz,1H),7.52(s,1H),7.24(t,J=8.4Hz,2H ),3.55(m,J=7.0,3.8Hz,1H),3.13(t,J=5.0Hz,4H),2.83(s,3H),2.65(q,J=7.4Hz ,2H),1.22(s,1H),1.16(t,J=6.7Hz,3H),0.95-0.83(m,2H),0.78(d,J=4.2Hz,2H).

[0313] Example 32 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazole-2-amine

[0314]

[0315] The same procedure as in Example 1 was performed, except that N-methylpiperazine was used instead of N-ethylpiperazine in step 1, and p-aminobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 194-196°C. ESI-HRMS (m / z): calcd. for C 29 H 30 FN8S[M+H] + ,541.2298,found 541.2298. 1H NMR(400MHz, DMSO-d6)δ8.10(t,J=7.2Hz,1H),7.80(s,1H),7.67(s,1H),7.47-7.32(m, 3H),7.04(s,2H),6.95(d,J=8.6Hz,1H),6.64(d,J=10.0Hz,1H),6.37(d,J=8.3Hz,1H),5 .78(s,1H),5.46(s,1H),3.46(q,J=7.0Hz,1H),2.96(d,J=22.6Hz,2H),2.32(d,J=8.3H z,1H),2.21(s,1H),2.15(s,4H),1.93(s,3H),1.08(t,J=7.0Hz,2H),0.79-0.56(m,2H).

[0316] Example 33 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0317]

[0318] The same procedure as in Example 1 was performed, except that N-methylpiperazine was used instead of N-ethylpiperazine in step 1, and p-cyanobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 166-167°C. ESI-HRMS (m / z): calcd. for C 30 H 28 FN8S[M+H] + ,551.2142,found 551.2141. 1 H NMR (400MHz, CDCl3) δ8.84(s,1H),7.99(t,J=8.9Hz,5H),7.78(d,J=8.2Hz,2H),7.54(s,1H),7. 30(s,2H),3.57(s,1H),3.29(s,4H),3.11(s,4H),2.71(s,3H),1.24-1.19(m,2H),0.79(s,2H).

[0319] Example 34 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0320]

[0321] The same procedure as in Example 1 was performed, except that N-methylpiperazine was substituted for N-ethylpiperazine in step 1, and p-nitrobenzaldehyde was substituted for benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange-red solid. Melting point: 178-179°C. ESI-HRMS (m / z): calcd. for C 29 H 28 FN8O2S[M+H] + ,571.2040,found 571.2034. 1 H NMR (400MHz, CDCl3) δ8.36(d,J=8.3Hz,2H),8.07(d,J=8.4Hz,2H),8.03-7.94(m,3H),7.78(s,1H),7.52(s,1H) ,7.16(s,2H),3.76(q,J=7.1Hz,1H),3.36(s,4H),3.13(s,4H),2.85(s,3H),1.20(d,J=6.9Hz,2H),0.80(s,2H).

[0322] Example 35 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0323]

[0324] The same operation as in Example 1 was performed, except that N-methylpiperazine was used instead of N-ethylpiperazine in step 1, and o-trifluoromethylbenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-trifluoromethylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 150-150.5°C. ESI-HRMS (m / z): calcd. for C 30 H 28 F4N7S[M+H] + ,594.2063,found594.2063. 1H NMR(400MHz,Chloroform-d)δ8.82(s,1H),8.00-7.92(m,3H),7.86(d,J=7.7Hz,1H),7.71(m,J=27.1,12.8,6.7Hz,3H),7.51(s,1H),7 .24(t,J=8.4Hz,2H),3.55(m,J=7.0,3.3Hz,1H),3.20(s,4H),2.94(s,4H),2.59(s,3H),1.18(d,J=6.7Hz,2H),0.78(d,J=3.8Hz,2H).

[0325] Example 36 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-hydroxyphenyl)-1,3,4-thiadiazole-2-amine

[0326]

[0327] The same procedure as in Example 1 was performed, except that N-methylpiperazine was used instead of N-ethylpiperazine in step 1 and hydroxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-hydroxyphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 270-272°C. ESI-HRMS (m / z): calcd. for C 29 H 29 FN7OS[M+H] + ,542.2138,found 542.2134. 1 H NMR (400MHz, DMSO-d6) δ9.80(s,1H),9.48(s,1H),8.21(s,1H),8.12-7.99(m,2H),7.45-7.36(m,3H),7.29(d,J=9.9Hz,2H),7.22 (d,J=7.7Hz,1H),6.88(d,J=8.1Hz,1H),3.80(s,1H),3.00(s,4H),2.65(s,4H),2.33(s,3H),1.15(d,J=6.7Hz,2H),0.66(s,2H).

[0328] Example 37 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-chlorophenyl)-1,3,4-thiadiazole-2-amine

[0329] Step 1: 1-cyclopropyl-6-N-methylpiperazinyl-2-(4-fluorophenyl)-5-aminobenzimidazole

[0330]

[0331] Compound 1 (1 mmol, 204.1 mg) was dissolved in 10 mL of tetrahydrofuran, and DIPEA (1.1 mmol) was added. A solution of cyclopropylamine (1.0 mmol) in 2 mL of tetrahydrofuran was then slowly added dropwise. The mixture was allowed to react at room temperature for 30 minutes. DIPEA (1.1 mmol) was then slowly added dropwise to a solution of N-methylpiperazine (1.1 mmol) in 2 mL of tetrahydrofuran. The reaction was allowed to react at room temperature. After the reaction was complete, a saturated ammonium chloride solution was added to the reaction solution, followed by extraction with dichloromethane and concentration to obtain the monosubstituted product. The disubstituted intermediate (1 mmol) was dissolved in a mixture of 5 mL of ethanol and 5 mL of tetrahydrofuran. A 1 mol / L aqueous potassium hydroxide solution was slowly added with stirring until a yellow precipitate formed. Stirring was continued for 10 minutes. The reaction solution was filtered, washed with copious amounts of water, and dried to obtain the disubstituted intermediate. It (1 mmol) was dissolved in a mixed solution of 5 mL of ethanol and 5 mL of tetrahydrofuran, and 1.0 eq of 10% Pd-C and 7.5 eq of HCOONH4 were added with stirring, and stirred at room temperature for 1 hour. The reaction solution was filtered into a 5 mL tetrahydrofuran solution containing 1.2 mmol of m-fluorobenzaldehyde, 1 mL of acetic acid was added, and stirred at room temperature. HPLC-MS detection was performed until the reaction was complete, and the reaction was concentrated under reduced pressure. The pH was adjusted to neutral with saturated sodium bicarbonate solution, and extracted with dichloromethane. Column chromatography was performed, and ethyl acetate-petroleum ether was used as the eluent to obtain the compound 1-cyclopropyl-6-N-methylpiperazinyl-2-(4-fluorophenyl)-5-aminobenzimidazole as a brown solid. ESI-MS (m / z): calcd.for C 21 H 25 FN5[M+H] + ,366.21,found 366.19.

[0332] Step 2: 1-cyclopropyl-6-N-methylpiperazinyl-2-(4-fluorophenyl)-5-thiosemicarbazide benzimidazole

[0333]

[0334] The same method as in Example 1 was used to obtain 1-cyclopropyl-6-N-methylpiperazinyl-2-(4-fluorophenyl)-5-thiosemicarbazide-benzimidazole as a white solid. ESI-HRMS (m / z): calcd.for C 22 H 27 FN7S[M+H] + ,440.20,found 440.26.

[0335] Step 3: N-(1-cyclopropyl-6-N-methylpiperazinyl-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0336]

[0337] The same operation as in Example 1 was performed, except that o-chlorobenzaldehyde was used in place of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-chlorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 199-201°C. ESI-HRMS (m / z): calcd. for C 29 H 28 ClFN7S[M+H] + ,560.1799,found560.1797. 1 H NMR (400MHz, CDCl3) δ8.79(s,1H),8.29(dd,J=7.3,2.8Hz,1H),8.04(d,J=2.2Hz,1H),7.96(dd,J=8.5,5.3Hz,2H),7.51(d,J=8.4Hz,2 H),7.41(dd,J=6.8,3.3Hz,2H),7.15(s,2H),3.55(s,1H),3.15(s,4H),2.87(s,4H),2.56(s,3H),1.18(d,J=6.8Hz,2H),0.78(s,2H).

[0338] Example 38 N-(1-cyclopropyl-6-ethoxy-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(3-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0339]

[0340] The procedure was the same as in Example 37. In step 1, the disubstituted product was obtained by replacing the tetrahydrofuran solution with a mixed solution of ethanol and tetrahydrofuran, and replacing DIPEA with an aqueous potassium hydroxide solution. In step 3, the disubstituted product was obtained by replacing o-chlorobenzaldehyde with chlorobenzaldehyde to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-chlorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 208-210°C. ESI-HRMS (m / z): calcd. for C 29 H 28 ClFN7S[M+H] + ,560.1799,found 560.1796.1 H NMR(400MHz,Chloroform-d)δ8.04-7.95(m,3H),7.94(s,1H),7.79(s,1H),7.76-7.71(m,1H),7.50(s,1H),7.42(d,J=5. 2Hz,2H),7.25(t,J=8.4Hz,2H),3.56(s,1H),3.16(s,4H),2.89(s,4H),2.56(s,3H),1.19(d,J=6.8Hz,2H),0.79(s,2H).

[0341] Example 39 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-chlorophenyl)-1,3,4-thiadiazole-2-amine

[0342]

[0343] The procedure was the same as in Example 37, except that in step 1, a mixed solvent of methanol and tetrahydrofuran was used instead of tetrahydrofuran, and potassium hydroxide aqueous solution was used instead of DIPEA to obtain a disubstituted product. In step 3, p-chlorobenzaldehyde was used instead of o-chlorobenzaldehyde to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-chlorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 163-164°C. ESI-HRMS (m / z): calcd. for C 29 H 28 ClFN7S[M+H] + ,560.1799,found 560.1796. 1 H NMR (400MHz, CDCl3) δ8.74(s,1H),8.03-7.93(m,3H),7.84(d,J=8.2Hz,2H),7.52-7.43(m,3H),7.25(t,J=8.4Hz ,2H),3.57(d,J=6.6Hz,1H),3.12(t,J=4.8Hz,4H),2.83(s,4H),2.53(s,3H),1.18(t,J=6.6Hz,2H),0.79(s,2H).

[0344] Example 40 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-bromophenyl)-1,3,4-thiadiazole-2-amine

[0345]

[0346] The same procedure as in Example 37 was performed, except that o-chlorobenzaldehyde was replaced with m-bromobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-bromophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 210-211°C. ESI-HRMS (m / z): calcd. for C 29 H 28 BrFN7S[M+H] + ,604.1294,found604.1293. 1 H NMR (400MHz, CDCl3) δ8.08(s,1H),8.02(s,1H),7.97(t,J=7.0Hz,2H),7.84-7.74(m,2H),7.57(d,J=8.2Hz,1H),7.49(s,1H),7.34(t ,J=7.9Hz,1H),7.16(s,2H),3.56(s,1H),3.24-3.05(m,4H),2.88(s,4H),2.54(d,J=9.2Hz,3H),1.18(d,J=6.7Hz,2H),0.78(s,2H).

[0347] Example 41 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-bromophenyl)-1,3,4-thiadiazole-2-amine

[0348]

[0349] The same procedure as in Example 37 was performed, except that p-bromobenzaldehyde was used in place of o-chlorobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-bromophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 170-171°C. ESI-HRMS (m / z): calcd. for C 29 H 28 BrFN7S[M+H] + ,604.1294,found604.1297. 1H NMR (400MHz, CDCl3) δ8.75(s,1H),8.03-7.92(m,3H),7.76(d,J=8.3Hz,3H),7.62(d,J=8.2Hz,2H),7.50(s,1H),7.25(t,J=8.5Hz,2 H),3.63-3.50(m,1H),3.18(s,4H),2.94(s,4H),2.59(s,3H),1.18(t,J=6.6Hz,2H),0.89(d,J=15.3Hz,1H),0.78(d,J=3.9Hz,2H).

[0350] Example 42 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0351]

[0352] The same procedure as in Example 37 was performed, except that o-fluorobenzaldehyde was used in place of o-chlorobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 202-203°C. ESI-HRMS (m / z): calcd. for C 29 H 28 F2N7S[M+H] + ,544.2095,found544.2095. 1 H NMR (400MHz, CDCl3) δ8.79(s,1H),8.33(t,J=7.6Hz,1H),8.04(s,1H),7.97(dd,J=8.4,5.3Hz,2H),7.47(m,2H),7.29-7.20( m,3H),7.17(s,1H),3.55(dd,J=7.0,3.6Hz,1H),3.24(s,4H),3.04(s,4H),2.65(s,3H),0.90(d,J=7.4Hz,2H),0.78(s,2H).

[0353] Example 43 N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazole-2-amine

[0354]

[0355] The same procedures as in Example 1 were performed, except that p-trifluoromethoxybenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1, and 4-aminobenzaldehyde was used instead of benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazol-2-amine as a pale green solid. Melting point: 179-180°C. ESI-HRMS (m / z): calcd. for C 31 H 32 F3N8OS[M+H] + ,621.2372,found621.2375. 1 H NMR(400MHz,DMSO-d6)δ8.27-8.05(m,3H),7.82(s,1H),7.55(dd,J=29.8,8.3Hz,3 H),7.38(d,J=20.7Hz,1H),7.02-6.90(m,2H),6.37(d,J=8.3Hz,1H),3.79(d,J=48 .5Hz,1H),3.46(d,J=7.2Hz,2H),2.95(s,4H),2.69(s,2H),2.31(d,J=7.1Hz,2H), 2.21(s,2H),1.07(d,J=6.9Hz,3H),0.98(t,J=7.2Hz,2H),0.65(d,J=26.9Hz,2H).

[0356] Example 44 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0357]

[0358] The procedure was the same as in Example 37, except that in step 1, a mixed solvent of methanol and tetrahydrofuran was used instead of tetrahydrofuran, and potassium hydroxide aqueous solution was used instead of DIPEA to obtain a disubstituted product. In step 3, 2-nitrobenzaldehyde was used instead of o-chlorobenzaldehyde to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 190-191°C. ESI-HRMS (m / z): calcd. for C 29 H 28 FN8O2S[M+H] + ,571.2040,found 571.2034. 1H NMR (400MHz, CDCl3) δ8.84(s,1H),7.98(q,J=4.6Hz,4H),7.84(d,J=7.6Hz,1H),7.78-7.70(m,2H),7.66(t,J=7.8Hz,1H),7 .50(s,1H),7.15(s,1H),3.56(m,1H),3.12(t,J=4.7Hz,4H),2.82(s,4H),2.51(s,3H),1.18(d,J=6.7Hz,2H),0.79(s,2H).

[0359] Example 45 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0360]

[0361] The same procedure as in Example 37 was performed, except that o-chlorobenzaldehyde was replaced with m-nitrobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange solid. Melting point: 110-111°C. ESI-HRMS (m / z): calcd. for C 29 H 28 FN8O2S[M+H] + ,571.2040,found571.2038. 1 H NMR(400MHz, CDCl3)δ8.70(s,1H),8.25(dd,J=29.1,8.1Hz,2H),8.04(s,1H),8.02-7.95(m,2H),7.67(t,J=8.0Hz,1H),7.51(s,1H), 7.26(t,J=8.3Hz,2H),7.17(s,1H),3.57(s,1H),3.21(d,J=5.8Hz,4H),3.00(s,4H),2.62(s,3H),1.20(d,J=6.8Hz,2H),0.79(s,2H).

[0362] Example 46 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-propoxyphenyl)-1,3,4-thiadiazole-2-amine

[0363]

[0364] The same procedure as in Example 37 was performed, except that para-propoxybenzaldehyde was used in place of o-chlorobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-propoxyphenyl)-1,3,4-thiadiazol-2-amine as a white solid. Melting point: 207-208°C. ESI-HRMS (m / z): calcd. for C 32 H 35 FN7OS[M+H] + ,584.2608,found 584.2608. 1 H NMR (400MHz, CDCl3) δ7.97(s,3H),7.77(d,J=7.9Hz,2H),7.52(s,1H),7.29(d,J=4.3Hz,3H),6.97(d,J=8.1Hz,2H),4.00(s,2H ),3.81(d,J=38.9Hz,1H),3.55(s,2H),3.34(s,6H),2.84(s,3H),1.46(s,2H),1.22(s,2H),1.08(t,J=7.1Hz,3H),0.77(s,2H).

[0365] Example 47 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-ethynylphenyl)-1,3,4-thiadiazole-2-amine

[0366]

[0367] The same procedure as in Example 37 was performed, except that p-ethynylbenzaldehyde was used instead of o-chloromethylbenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-ethynylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 247-248°C. ESI-HRMS (m / z): calcd. for C 31 H 29 FN7S[M+H] + ,550.2189,found 550.2189. 1H NMR (400MHz, CDCl3) δ8.73(s,1H),7.97(t,J=5.0Hz,3H),7.89-7.81(m,2H),7.60(d,J=7.9Hz,2H),7.52(s,1H),7.30( d,J=2.7Hz,2H),3.56(s,1H),3.31(s,3H),3.24(s,2H),3.16(s,3H),2.73(s,3H),1.21(d,J=6.7Hz,3H),0.78(s,2H).

[0368] Example 48 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3,4-ethylenedioxyphenyl)-1,3,4-thiadiazole-2-amine

[0369]

[0370] The procedure was the same as in Example 37, except that 3,4-ethylenedioxybenzaldehyde was used instead of o-chlorobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3,4-ethylenedioxyphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 160-161°C. ESI-HRMS (m / z): calcd. for C 31 H 31 FN7O2S[M+H] + ,584.2244,found 584.2244. 1 H NMR (400MHz, CDCl3) δ7.97(d,J=8.6Hz,3H),7.49(s,1H),7.43(d,J=2.4Hz,1H),7.35(d,J=8.5Hz,1H),7.30(s,1H),7.26(d,J=8.6Hz,1H) ,7.17(s,1H),6.96(d,J=8.3Hz,1H),4.34(s,4H),3.56(s,1H),3.22(s,4H),3.01(s,4H),2.63(s,3H),1.19(d,J=6.8Hz,2H),0.78(s,2H).

[0371] Example 49 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-formamidophenyl)-1,3,4-thiadiazole-2-amine

[0372]

[0373] The same procedure as in Example 37 was performed, except that in step 3, p-acetamidobenzaldehyde was used instead of o-chlorobenzaldehyde to obtain N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-formamidophenyl)-1,3,4-thiadiazol-2-amine as a white solid. Melting point: 228-230°C. ESI-HRMS (m / z): calcd. for C 31 H 32 FN8OS[M+H] + ,583.2404,found 583.2404. 1 H NMR (400MHz, DMSO-d6) δ10.26(s,1H),8.29(s,1H),8.11-8.03(m,2H),7.90-7.69(m,3H),7.50(d,J=8.5Hz,1H),7.41(p,J=9 .3Hz,4H),3.82(m,1H),3.10(s,5H),2.98(s,3H),2.10(s,2H),2.03(s,1H),1.93(s,2H),1.15(d,J=6.8Hz,3H),0.66(s,2H).

[0374] Example 50 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-iodophenyl)-1,3,4-thiadiazole-2-amine

[0375]

[0376] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and m-iodobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-iodophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 220-221°C. ESI-HRMS (m / z): calcd. for C 30 H 30 FIN7S[M+H] + ,666.1312,found 666.1313. 1H NMR (400MHz, CDCl3) δ8.75(s,1H),8.29(s,1H),8.01(s,1H),7.78(dq,J=13.3,7.5Hz,3H),7.56(d,J=9.3Hz,2H),7.35(t,J=7.5Hz,1 H),7.22(t,J=8.3Hz,1H),3.56(s,1H),3.18(s,4H),2.92(s,4H),2.74(s,2H),1.46(s,2H),1.03(d,J=7.0Hz,3H),0.72-0.66(m,2H).

[0377] Example 51 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-iodophenyl)-1,3,4-thiadiazole-2-amine

[0378]

[0379] The same operation as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1 and o-iodobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-iodophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 115-116°C. ESI-HRMS (m / z): calcd. for C 30 H 30 FIN7S[M+H] + ,666.1312,found 666.1314. 1 H NMR (400MHz, CDCl3) δ8.84(s,1H),8.04(d,J=6.6Hz,2H),7.85(d,J=7.8Hz,1H) ,7.75(t,J=7.5Hz,1H),7.56(s,2H),7.49(t,J=7.7Hz,1H),7.34(t,J=7.6Hz,1 H),7.25(d,J=9.4Hz,1H),7.17(t,J=7.7Hz,1H),3.56(s,1H),3.18(s,4H),2.7 2(s,2H),1.02(d,J=6.9Hz,3H),0.90(d,J=11.7Hz,3H),0.69(d,J=3.9Hz,2H).

[0380] Example 52 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-iodophenyl)-1,3,4-thiadiazole-2-amine

[0381]

[0382] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and p-iodobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-iodophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 118-120°C. ESI-HRMS (m / z): calcd. for C 30 H 30 FIN7S[M+H] + ,666.1312,found 666.1314. 1 H NMR (400MHz, CDCl3) δ7.96 (s, 1H), 7.84 (d, J = 7.9Hz, 2H), 7.75 (t, J = 7.2Hz, 2H), 7.66-7.52 (m, 4H), 7.36 (t, J = 8.0 Hz,2H),7.25(s,1H),3.68(s,2H),3.56(s,3H),3.14(s,3H),2.05(s,4H),1.11(s,3H),0.99(s,2H),0.69(s,2H).

[0383] Example 53 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-hydroxyphenyl)-1,3,4-thiadiazole-2-amine

[0384]

[0385] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and p-hydroxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-hydroxyphenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 238-240°C. ESI-HRMS (m / z): calcd. for C 30 H 31 FN7OS[M+H] + ,556.2295,found 556.2295. 1H NMR (400MHz, CDCl3) δ7.77(d,J=13.8Hz,2H),7.59(d,J=10.6Hz,4H),7.38(t,J=7.9Hz,1H),7.30(d,J=2.0Hz,3H),7.05 (d,J=8.6Hz,2H),3.77(s,1H),3.56(s,3H),3.32(s,2H),3.17(s,5H),1.09(d,J=7.2Hz,3H),0.92(s,2H),0.68(s,2H).

[0386] Example 54 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-ethoxyacylphenyl)-1,3,4-thiadiazole-2-amine

[0387]

[0388] The same procedure as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1 and p-carboxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-carboxyphenyl)-1,3,4-thiadiazole-2-amine as an orange solid. Melting point: 189-191°C. ESI-HRMS (m / z): calcd. for C 31 H 31 FN7O2S[M+H] + ,612.2557,found 612.2568. 1 H NMR (400MHz, CDCl3) δ8.70 (s, 1H), 7.97 (s, 1H), 7.92 (d, J = 7.5Hz, 1H), 7.74 (t, J=7.4Hz,1H),7.69-7.51(m,5H),7.34(t,J=7.5Hz,1H),7.25(d,J=9.6Hz,1H),4 .33(q,J=7.1Hz,2H),3.55(s,1H),3.27(s,5H),3.03(s,3H),2.81(s,2H),1.73( s,3H),1.03(d,J=6.8Hz,3H),0.98(d,J=6.3Hz,2H),0.69(q,J=3.9,2.6Hz,2H).

[0389] Example 55 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-ethoxyacylphenyl)-1,3,4-thiadiazole-2-amine

[0390]

[0391] The same procedure as in Example 1 was performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and m-carboxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-carboxyphenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 191-193°C. ESI-HRMS (m / z): calcd. for C 31 H 31 FN7O2S[M+H] + ,612.2557,found 612.2568. 1 H NMR (400MHz, CDCl3) δ8.60 (s, 1H), 8.47 (s, 1H), 8.37 (d, J = 7.8Hz, 1H), 8.11 (d ,J=6.8Hz,2H),7.78(q,J=7.2Hz,1H),7.66-7.55(m,3H),7.38(t,J=7.5Hz,1H ),4.41(dq,J=28.8,7.1Hz,2H),3.95(t,J=5.3Hz,1H),3.85(s,1H),3.68(s,3 H),3.62-3.47(m,4H),3.27-3.16(m,2H),1.09(d,J=7.3Hz,3H),0.69(s,2H).

[0392] Example 56 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-methanesulfonylaminophenyl)-1,3,4-thiadiazole-2-amine

[0393]

[0394] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and m-methylsulfonamidobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-methylsulfonamidophenyl)-1,3,4-thiadiazol-2-amine as a white solid. Melting point: 247-249°C. ESI-HRMS (m / z): calcd. for C 31 H 34 FN8O2S2[M+H] + ,633.2230,found 633.2231. 1H NMR (400MHz, CDCl3) δ11.92(s,1H),9.43(s,1H),8.94(s,1H),7.98(s,1H),7.75(d,J=7.9Hz,2H),7.55(d,J=31.2Hz,4H),7.36(d d,J=15.5,7.8Hz,2H),3.76(s,1H),3.68(s,1H),3.57(s,6H),3.39-3.20(m,4H),3.15(s,4H),1.09(d,J=6.9Hz,3H),0.70(s,2H).

[0395] Example 57 N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0396]

[0397] The same procedures as in Example 37 were performed, except that m-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1, and p-nitrobenzaldehyde was used instead of o-chlorobenzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as a reddish-brown solid. Melting point: 206-208°C. ESI-HRMS (m / z): calcd. for C 29 H 28 FN8O2S[M+H] + ,571.2040,found 571.2041. 1 H NMR (400MHz, Chloroform-d) δ8.37(d,J=8.4Hz,2H),8.07(d,J=8.4Hz,2H),7.98(s,1H),7.80(d,J=7.8Hz,1H),7.72(d,J=9.7H z,1H),7.61-7.51(m,2H),7.30(s,3H),3.76(q,J=7.0Hz,1H),3.60(s,2H),3.30(s,6H),2.82(s,3H),1.46(s,2H),0.81(s,2H).

[0398] Example 58 N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0399]

[0400] The same procedures as in Example 1 were performed, except that o-fluorobenzaldehyde was used instead of p-fluorobenzaldehyde in step 1 and p-nitrobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange-red solid. Melting point: 241-242°C. ESI-HRMS (m / z): calcd. for C 30 H 30 FN8O2S[M+H] + ,585.2196,found 585.2196. 1 H NMR (400MHz, CDCl3) δ8.35(d,J=8.3Hz,1H),8.08-7.97(m,2H),7.75(d,J=7.9Hz,1H),7.59(d,J=7.5Hz,2H),7.38(d,J=7.0Hz,1H ),7.29(s,4H),3.84(s,2H),3.65(s,2H),3.55(s,1H),3.39(s,2H),3.22(s,2H),2.96(s,3H),1.09(d,J=6.8Hz,2H),0.69(s,2H).

[0401] Example 59 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-isopropylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0402]

[0403] The same procedures as in Example 1 were performed, except that N-isopropylpiperazine was substituted for N-ethylpiperazine in Step 1, and p-nitrobenzaldehyde was substituted for benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-isopropylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as a yellow-brown solid. Melting point: 221-222°C. ESI-HRMS (m / z): calcd. for C 31 H 32 FN8O2S[M+H] + ,599.2353,found 599.2353. 1H NMR (400MHz, DMSO-d6) δ11.01(s,1H),10.18(s,1H),8.54(s,1H),8.35(d,J=9.3Hz,2H),8.11(m,4H),7.4 4(d,J=12.8Hz,3H),3.86(s,1H),3.51(s,1H),3.48-3.36(m,8H),1.23(s,6H),0.86(s,2H),0.70(s,2H).

[0404] Example 60 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0405]

[0406] The same procedure as in Example 1 was performed, except that N-acetylpiperazine was substituted for N-ethylpiperazine in step 1, and 3,4,5-trimethoxybenzaldehyde was substituted for benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 147-148°C. ESI-HRMS (m / z): calcd. for C 33 H 35 FN7O4S[M+H] + ,644.2455,found644.2455. 1 H NMR(400MHz,DMSO-d6)δ9.63(s,1H),8.28(s,1H),8.06(dd,J=8.5,5.5Hz,2H),7.47-7.37(m,3H),7.10(s,2H),3.88(s,6H),3 .79(s,1H),3.74(s,3H),3.67(d,J=9.2Hz,4H),3.03-2.86(m,4H),2.08(s,3H),1.14(d,J=6.9Hz,2H),0.66(d,J=4.0Hz,2H).

[0407] Example 61 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0408]

[0409] The same procedures as in Example 1 were performed, except that N-acetylpiperazine was substituted for N-ethylpiperazine in step 1, and p-nitrobenzaldehyde was substituted for benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as a reddish-brown solid. Melting point: 147-148°C. ESI-HRMS (m / z): calcd. for C 30 H 28 FN8O3S[M+H] + ,599.1989,found 599.1989. 1 H NMR(400MHz,Chloroform-d)δ8.89(s,1H),8.36(d,J=8.4Hz,2H),8.04(m,5H),7.44(s,1H),7.26(d,J=8.4Hz,2H ),3.94(s,2H),3.79(s,2H),3.60(s,1H),3.03(q,J=5.7Hz,4H),2.23(s,3H),1.23(d,J=6.9Hz,2H),0.82(s,2H).

[0410] Example 62 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0411]

[0412] The same procedure as in Example 1 was performed, except that N-acetylpiperazine was substituted for N-ethylpiperazine in step 1, and p-fluorobenzaldehyde was substituted for benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 149-150°C. ESI-HRMS (m / z): calcd. for C 30 H 28 F2N7OS[M+H] + ,572.2044,found 572.2231. 1H NMR (400MHz, DMSO-d6) δ9.67(s,1H),8.35(s,1H),8.06(t,J=6.8Hz,2H),7.91(t,J=6.7Hz,2H),7.39(m,5H ),3.79(s,1H),3.68(d,J=8.9Hz,4H),3.03-2.85(m,4H),2.08(s,3H),1.14(d,J=6.8Hz,2H),0.66(s,2H).

[0413] Example 63 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0414]

[0415] The same procedure as in Example 1 was performed, except that N-acetylpiperazine was substituted for N-ethylpiperazine in step 1, and p-cyanobenzaldehyde was substituted for benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine as an orange solid. Melting point: 147-149°C. ESI-HRMS (m / z): calcd. for C 31 H 28 FN8OS[M+H] + ,579.2091,found 579.2089. 1 H NMR(400MHz, DMSO-d6)δ9.87(s,1H),8.32(d,J=6.5Hz,1H),8.14-7.93(m,6H),7.40(d,J=9.9H z,3H),3.79(s,1H),3.67(s,4H),2.95(s,4H),2.08(s,3H),1.14(d,J=6.8Hz,2H),0.66(s,2H).

[0416] Example 64 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0417]

[0418] The same procedure as in Example 1 was performed, except that N-acetylpiperazine was substituted for N-ethylpiperazine in step 1, and p-trifluoromethylbenzaldehyde was substituted for benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 145-147°C. ESI-HRMS (m / z): calcd. for C 31 H 28 F4N7OS[M+H] + ,622.2012,found 622.2014. 1 H NMR (400MHz, DMSO-d6) δ9.83(s,1H),8.34(s,1H),8.08(d,J=7.6Hz,4H),7.88(d,J=8.1Hz,2H),7.42(q,J =8.3,6.5Hz,3H),3.80(s,1H),3.68(s,4H),2.95(s,4H),2.08(s,3H),1.14(d,J=6.8Hz,2H),0.67(s,2H).

[0419] Example 65 N-(1-isopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0420]

[0421] The same procedures as in Example 37 were performed, except that isopropylamine was used instead of cyclopropylamine in Step 1, and p-nitrobenzaldehyde was used instead of o-chlorobenzaldehyde in Step 3 to obtain N-(1-isopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazol-2-amine as a reddish-brown solid. Melting point: 153-155°C. ESI-HRMS (m / z): calcd. for C 29 H 30 FN8O2S[M+H] + ,573.2196,found 573.2197. 1H NMR(400MHz, DMSO-d6)δ9.83(s,1H),8.34(dd,J=8.9,2.3Hz,2H),8.18(s,1H),8.15-8.09(m,2H),7.73(m,J=6.8,5.4,2.4H z,2H),7.52-7.39(m,3H),4.68(dd,J=13.8,6.9Hz,1H),3.02(s,4H),2.60(s,4H),2.30(s,3H),1.62(dd,J=6.9,2.2Hz,6H).

[0422] Example 66 N-(1-pentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0423]

[0424] The same procedures as in Example 37 were performed, except that pentylamine was used instead of cyclopropylamine in Step 1 and p-nitrobenzaldehyde was used instead of o-chlorobenzaldehyde in Step 3 to obtain N-(1-pentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazol-2-amine as a reddish-brown solid. Melting point: 120-122°C. ESI-HRMS (m / z): calcd. for C 31 H 34 FN8O2S[M+H] + ,601.2509,found 601.2509. 1 H NMR (400MHz, DMSO-d6) δ9.81 (s, 1H), 8.31 (d, J = 8.4Hz, 2H), 8.15 (s, 1H), 8.09 (d, J = 8.4Hz, 2H), 7.81 (dd, J = 8.3, 5.4Hz, 2H), 7.43 (dd, J = 17.1, 8.4Hz,3H),4.29(t,J=7.4Hz,2H),2.98(t,J=4.7Hz,4H),2.57(s,4H),2.27(s,3H),1.65(p,J=7.2Hz,2H),1.15(m,4H),0.75(t,J=6.9Hz,3H).

[0425] Example 67 N-(1-cyclopentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0426]

[0427] The same procedure as in Example 37 was performed, except that cyclopentylamine was substituted for cyclopropylamine in step 1, and p-nitrobenzaldehyde was substituted for o-chlorobenzaldehyde in step 3 to obtain N-(1-cyclopentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange solid. Melting point: 251-253°C. ESI-HRMS (m / z): calcd. for C 31 H 32 FN8O2S[M+H] + ,599.2353,found 599.2350. 1 H NMR (400MHz, DMSO-d6) δ9.84(s,1H),8.34(d,J=8.0Hz,2H),8.20(s,1H),8.12(d,J=8.3Hz,2H),7.74(s,2H),7.45(d,J=11.0H z,2H),7.31(s,1H),4.85(s,1H),3.00(s,4H),2.29(s,2H),2.22(s,2H),2.13(s,2H),2.00(s,2H),1.73(s,3H),0.89(s,4H).

[0428] Example 68 N-(1-cyclopropyl-2-phenyl-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0429]

[0430] The same procedures as in Example 37 were performed, except that benzaldehyde was used instead of p-fluorobenzaldehyde in Step 1 and p-nitrobenzaldehyde was used instead of o-chlorobenzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(phenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazol-2-amine as a reddish-brown solid. Melting point: 192-193°C. ESI-HRMS (m / z): calcd. for C 29 H 29 N8O2S[M+H] + ,553.2134,found 553.2137. 1H NMR (400MHz, DMSO-d6) δ9.89(s,1H),8.35(d,J=8.5Hz,2H),8.24(s,1H),8.12(d,J=8.5Hz,2H),8.01(d,J=7.0Hz,2H),7.76-7.67(m,1H),7. 57(d,J=7.0Hz,2H),7.40(s,1H),3.83(d,J=7.5Hz,1H),3.11(s,4H),1.27(s,4H),1.14(d,J=6.9Hz,2H),0.88(d,J=6.4Hz,3H),0.66(s,2H).

[0431] Example 69 N-(1-cyclopropyl-2-(4-chlorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0432]

[0433] The same procedures as in Example 37 were performed, except that p-chlorobenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1, and p-nitrobenzaldehyde was used instead of o-chlorobenzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-chlorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as a reddish-brown solid. Melting point: 157-159°C. ESI-HRMS (m / z): calcd. for C 29 H 28 ClN8O2S[M+H] + ,587.1744,found 587.1747. 1 H NMR(400MHz,Chloroform-d)δ8.41-8.31(m,2H),8.10-8.03(m,2H),8.02-7.88(m,3H),7.58-7.52(m,2H),7. 30(d,J=3.0Hz,2H),3.58(s,1H),3.26(s,4H),3.03(s,4H),2.66(d,J=17.8Hz,3H),1.23(s,2H),0.80(s,2H).

[0434] Example 70 N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0435]

[0436] The same procedure as in Example 1 was performed, except that p-trifluoromethoxybenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1 and p-nitrobenzaldehyde was used instead of benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange solid. Melting point: 190-192°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F3N8O3S[M+H] + ,651.2114,found651.2108. 1 H NMR(400MHz, DMSO-d6)δ9.82(s,1H),8.32(d,J=8.5Hz,2H),8.24-8.03(m,5H),7.55(d,J=8.3Hz,2H),7.39(s,1H),3.81(dd, J=7.7,4.0Hz,1H),3.00(s,4H),2.64(s,4H),2.41(d,J=17.0Hz,2H),1.15(d,J=6.7Hz,2H),1.09-1.02(m,3H),0.67(s,2H).

[0437] Example 71 N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0438]

[0439] The same procedures as in Example 1 were performed, except that p-trifluoromethoxybenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1, and p-trifluoromethylbenzaldehyde was used instead of benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 222-224°C. ESI-HRMS (m / z): calcd. for C 32 H 30 F6N7OS[M+H] + ,674.2137,found674.2137. 1H NMR (400MHz, DMSO-d6) δ9.81(s,1H),8.39(s,1H),8.17(d,J=8.4Hz,2H),8.08(d,J=8.1Hz,2H),7.89(d,J=8.1Hz,2H),7.57( d,J=8.4Hz,2H),7.42(s,1H),3.84(s,1H),3.34(s,4H),3.22(s,4H),3.15-3.05(m,2H),1.19(t,J=8.4Hz,5H),0.70(s,2H).

[0440] Example 72 N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0441]

[0442] The same procedures as in Example 1 were performed, except that p-trifluoromethoxybenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1 and p-fluorobenzaldehyde was used instead of benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 127-129°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F4N7OS[M+H] + ,624.2169,found 624.2167. 1 H NMR (400MHz, DMSO-d6) δ9.64(s,1H),8.41(s,1H),8.16(d,J=8.3Hz,2H),7.92(s,2H),7.57(d,J=8.3Hz,2H) ,7.47-7.31(m,3H),3.84(s,1H),3.34(s,8H),3.21(s,2H),1.26(s,2H),1.17(d,J=6.9Hz,3H),0.70(s,2H).

[0443] Example 73 N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0444]

[0445] The same procedure as in Example 1 was performed, except that p-trifluoromethoxybenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and p-cyanobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanomethylphenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 134-136°C. ESI-HRMS (m / z): calcd. for C 32 H 30 F3N8OS[M+H] + ,631.2215,found631.2216. 1 H NMR(400MHz, DMSO-d6)δ9.78(s,1H),8.22-8.12(m,3H),8.03(d,J=8.1Hz,2H),7.97(d,J=8.2Hz,2H),7.56(d,J=8.3Hz,2H), 7.41(s,1H),3.82(s,1H),3.01(s,4H),2.63(s,4H),2.44(s,2H),1.17(d,J=6.8Hz,2H),1.06(t,J=7.0Hz,3H),0.69(s,2H).

[0446] Example 74 N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0447]

[0448] The same procedure as in Example 1 was performed, except that p-trifluoromethoxybenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and 3,4,5-trimethoxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazol-2-amine as a white solid. Melting point: 264-266°C. ESI-HRMS (m / z): calcd. for C 34 H 37 F3N7O4S[M+H] + ,696.2580,found 696.2582. 1HNMR(400MHz,DMSO-d6)δ9.51(s,1H),8.27-8.07(m,3H),7.56(d,J=8.3Hz,2H),7.41(s,1H),7.10(s,2H),3.87(s,6H),3.82(s,1 H),3.73(s,3H),3.02(s,4H),2.69(s,4H),2.50-2.23(m,2H),1.17(d,J=6.8Hz,2H),1.09(t,J=7.3Hz,3H),0.69(d,J=3.8Hz,2H).

[0449] Example 75 N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0450]

[0451] The same procedure as in Example 1 was performed, except that p-trifluoromethoxybenzaldehyde was used in place of p-fluorobenzaldehyde in Step 1 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazol-2-amine as a white solid. Melting point: 116-118°C. ESI-HRMS (m / z): calcd. for C 31 H 31 F3N7OS[M+H] + ,606.2263,found606.2263. 1 H NMR (400MHz, DMSO-d6) δ9.70(s,1H),8.42(s,1H),8.16(d,J=8.4Hz,2H),7.85(d,J=7.1Hz,2H),7.53(m,J=14.6,7.8Hz,5H),7.40(s,1H),3.89 -3.79(m,1H),3.44(dd,J=14.2,7.0Hz,2H),3.21(s,6H),2.98(s,2H),1.26(q,J=6.2,5.5Hz,3H),1.16(d,J=6.7Hz,2H),0.69(d,J=3.7Hz,2H).

[0452] Example 76 N-(1-cyclopropyl-2-(4-fluorophenyl)-6-fluoro-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0453]

[0454] The procedure was the same as in Example 1, except that monosubstitution occurred in step 1 and benzaldehyde was replaced with m-nitrobenzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-fluoro-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 279-281°C. ESI-HRMS (m / z): calcd. for C 24 H 17 F2N6O2S[M+H] + ,491.1102,found 491.1099. 1 H NMR (400MHz, DMSO-d6) δ10.46(s,1H),8.62(s,1H),8.56(d,J=7.4Hz,1H),8.39-8.25(m,2H),8.09(dd,J=8.5,5.6Hz,2H ),7.83(t,J=8.0Hz,1H),7.65(d,J=10.8Hz,1H),7.43(t,J=8.7Hz,2H),3.82(s,1H),1.14(d,J=6.9Hz,2H),0.68(s,2H).

[0455] Example 77 N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine

[0456]

[0457] The same procedures as in Example 1 were performed, except that p-trifluoromethylbenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1 and p-nitrobenzaldehyde was used instead of benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine as an orange solid. Melting point: 257-259°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F3N8O2S[M+H] + ,635.2165,found 635.2165. 1H NMR (400MHz, DMSO-d6) δ9.89(s,1H),8.35(d,J=8.5Hz,2H),8.27(d,J=8.4Hz,3H),8.12(d,J=8.5Hz,2H),7.94(d,J=8.1Hz,2H),7.4 3(s,1H),3.88(s,1H),3.10(s,5H),2.76(s,3H),2.68(d,J=14.0Hz,2H),1.19(d,J=6.8Hz,3H),1.14(s,2H),0.70(d,J=3.8Hz,2H).

[0458] Example 78 N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine

[0459]

[0460] The same procedure as in Example 1 was performed, except that p-trifluoromethylbenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and p-cyanobenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine as a yellow solid. Melting point: 186-188°C. ESI-HRMS (m / z): calcd. for C 32 H 30 F3N8S[M+H] + ,615.2266,found 615.2266. 1 H NMR (400MHz, DMSO-d6) δ9.83(s,1H),8.32(s,1H),8.26(d,J=8.1Hz,2H),8.04(d,J=8.1Hz,2H),7.96(dd,J=15.6,8.0Hz,4H ),7.43(s,1H),3.87(s,1H),3.26-3.02(m,5H),2.89(s,3H),2.69(s,2H),1.25(s,2H),1.18(d,J=6.7Hz,3H),0.70(s,2H).

[0461] Example 79 N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine

[0462]

[0463] The same operation as in Example 1 was performed, except that p-trifluoromethylbenzaldehyde was used instead of p-fluorobenzaldehyde in step 1, and 3,4,5-trimethoxybenzaldehyde was used instead of benzaldehyde in step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 148-150°C. ESI-HRMS (m / z): calcd. for C 34 H 37 F3N7O3S[M+H] + ,680.2653,found 680.2653. 1 H NMR (400MHz, DMSO-d6) δ9.55(s,1H),8.26(d,J=7.8Hz,3H),7.93(d,J=8.1Hz,2H),7.43(s,1H),7.10(s,2H),3.88(s, 6H),3.73(s,3H),3.08(s,5H),2.79(s,3H),2.69(d,J=4.1Hz,2H),1.19(d,J=17.4Hz,3H),1.12(s,2H),0.69(s,2H).

[0464] Example 80 N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine

[0465]

[0466] The same procedures as in Example 1 were performed, except that p-trifluoromethylbenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1 and p-trifluoromethylbenzaldehyde was used instead of benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 159-161°C. ESI-HRMS (m / z): calcd. for C 32 H 30 F6N7S[M+H] + ,658.2188,found658.2188. 1H NMR(400MHz,DMSO-d6)δ9.78(s,1H),8.32(s,1H),8.26(s,2H),8.07(s,2H),7.94(s,2H),7.88(s,2H),7.43( s,1H),3.87(s,1H),3.11(s,4H),2.94(s,4H),2.73(s,2H),1.19(d,J=13.4Hz,3H),1.15(s,2H),0.70(s,2H).

[0467] Example 81 N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine

[0468]

[0469] The same procedure as in Example 1 was performed, except that p-trifluoromethylbenzaldehyde was used instead of p-fluorobenzaldehyde in Step 1 and p-fluorobenzaldehyde was used instead of benzaldehyde in Step 3 to obtain N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 161-163°C. ESI-HRMS (m / z): calcd. for C 31 H 30 F4N7S[M+H] + ,608.2220,found 608.2222. 1 H NMR (400MHz, DMSO-d6) δ9.61(s,1H),8.33(s,1H),8.26(d,J=8.0Hz,2H),7.92(dd,J=13.5,8.2Hz,4H),7.38( dd,J=19.3,10.7Hz,3H),3.87(s,1H),3.10(s,5H),2.92(s,3H),1.19(s,2H),1.18-1.12(m,3H),0.69(s,2H).

[0470] Example 82 N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine

[0471]

[0472] The same procedure as in Example 1 was performed, except that p-trifluoromethylbenzaldehyde was used in place of p-fluorobenzaldehyde in Step 1 to obtain N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazol-2-amine as a yellow solid. Melting point: 179-181°C. ESI-HRMS (m / z): calcd. for C 31 H 31 F3N7S[M+H] + ,590.2314,found590.2314. 1 H NMR (400MHz, DMSO-d6) δ9.63(s,1H),8.34(s,1H),8.26(d,J=8.0Hz,2H),7.94(d,J=8.0Hz,2H),7.85(d,J=7.0Hz,2H),7.52(d,J=7.7Hz,3H), 7.42(s,1H),3.87(s,1H),3.14(d,J=34.4Hz,5H),2.87(d,J=18.7Hz,3 H),2.68(d,J=3.8Hz,2H),1.19(s,2H),1.18-1.12(m,3H),0.69(s,2H).

[0473] Pharmacologically active part

[0474] Experimental Example 1: Evaluation of the inhibitory effect of the example compounds on the IL-6 / JAK / STAT3 signaling pathway

[0475] 1) Principle: Invivogen utilizes HEK-Blue IL-6 cells stably expressing the IL-6 receptor and a secreted embryonic alkaline phosphatase (SEAP) reporter gene whose promoter contains a STAT3 binding domain. Upon IL-6 stimulation, phosphorylated STAT3 in the cytoplasm forms a dimer, translocates into the nucleus, binds to the SEAP reporter gene, initiates transcription, and ultimately produces SEAP, which is secreted extracellularly. SEAP in the cell culture supernatant reacts with Quanti-Blue, and the amount of SEAP produced is calculated by the product absorbance at 655 nm, reflecting the activity of the IL6-STAT3 pathway. Therefore, after drug treatment of cells, the supernatant is reacted with Quanti-Blue, and the amount of SEAP produced is measured at 655 nm to evaluate the inhibitory effect of compounds on the IL-6-stimulated STAT3 signaling pathway.

[0476] 2) Experimental method: When the cell density reaches 80%, gently tap the attached cells, resuspend them in complete culture medium preheated at 37°C and count them. Use a sterile 96-well cell culture plate and add the cell suspension (the final cell number per well is 5×10 4 The system was prepared by adding 160 μL of the solution containing 100 μL of each cell suspension, 20 μL of the compound at different concentrations, and 20 μL of the IL-6 cytokine (final concentration of 1 ng / mL), for a total of 200 μL. The negative control group consisted of 40 μL of complete culture medium and 160 μL of cell suspension; the positive control group consisted of 20 μL of IL-6 cytokine (final concentration of 1 ng / mL), 20 μL of complete culture medium, and 160 μL of cell suspension. After incubation at 37°C for 16 hours, 20 μL of the solution was transferred from each well of the 96-well plate to another 96-well plate. 180 μL of QUANTI-Blue colorimetric solution preheated at 37°C was added. After incubation at 37°C for 1 hour, the cells were detected at a wavelength of 655 nm.

[0477] 3) The results showed that the Example compounds could inhibit the IL-6 / JAK / STAT3 pathway in a dose-dependent manner, and the experiment was reproducible. The inhibitory activities of some Example compounds are listed in Table 1.

[0478] Table 1. Inhibitory activity of the example compounds on the IL-6 / JAK / STAT3 signaling pathway in HEK-Blue cells.

[0479]

[0480]

[0481]

[0482] *: A indicates IC 50 ≤5μM, B means 5μM>IC 50 ≤10μM, C means 10μM>IC 50 ≤15μM, D means 15μM>IC 50 ≤20μM, E represents IC 50 >20μM.

[0483] Experimental Example 2: MTT assay to determine the inhibitory effect of the example compounds on tumor cells

[0484] 1) Experimental method: Breast cancer cell lines MDA-MB-231 and MDA-MB-468 in the logarithmic growth phase were digested with trypsin and prepared at a concentration of 2×10 4100 μL of cell suspension (2000 cells) was seeded into each well of a 96-well plate. The next day, 100 μL of fresh culture medium containing various drug concentrations and solvent control (final DMSO concentration <0.2%) was added to each well. Three parallel wells were set up for each group. Culture was continued at 37°C for 48 hours, and the supernatant was discarded. 50 μg / mL MTT solution was added to each well and incubated at 37°C for 4 hours. The supernatant was discarded, DMSO was added and mixed by vortexing, and the absorbance was measured at 570 nm using a microplate reader.

[0485] 2) The results showed that the example compounds inhibited MDA-MB-231 and MDA-MB-468 cells in a dose-dependent manner, and the experiment was reproducible. This indicates that the benzimidazolo-2-amino-1,3,4-thiadiazole derivatives possess antitumor activity. Some experimental data are listed in Tables 2 and 3.

[0486] Table 2. Screening results of the example compounds using MTT assay on MDA-MB-231 cells.

[0487]

[0488]

[0489]

[0490]

[0491] *: A indicates IC 50 ≤5μM, B means 5μM>IC 50 ≤10μM, C means 10μM>IC 50 ≤15μM, D means 15μM>IC 50 ≤20μM, E represents IC 50 >20μM.

[0492] Table 3. Screening results of the example compounds using MTT assay on MDA-MB-468 cells.

[0493]

[0494]

[0495]

[0496] *: A indicates IC 50 ≤5μM, B means 5μM>IC 50 ≤10μM, C means 10μM>IC 50 ≤15μM, D means 15μM>IC 50 ≤20μM, E represents IC 50 >20μM.

[0497] Experimental Example 3: Circular Dichroism Spectroscopy Evaluation of the Interaction between Example Compounds and STAT3 Protein

[0498] 1) Principle: STAT3 protein produces a specific circular dichroism signal at 190-260nm. When small molecule compounds bind to STAT3 protein, they cause the three-dimensional conformation of STAT3 protein to change, resulting in a change in the circular dichroism signal of STAT3 protein.

[0499] 2) Experimental Method: Prepare 0.01M phosphate buffer at pH 7.4. Use this phosphate buffer to prepare a 5.7 μM STAT3 protein solution. Incubate at 37°C in a water bath for 1 hour, and measure the circular dichroism spectrum of the STAT3 protein solution at 190-260 nm. Prepare a mixed solution of the example compound and STAT3 protein at a concentration of 28.5 μM and 5.7 μM, incubate at 37°C for 1 hour, and measure the circular dichroism spectrum of the mixture at 190-260 nm.

[0500] 3) The results showed that after the Example compounds were co-incubated with STAT3 protein, the circular dichroism spectra of STAT3 protein showed significant differences, indicating that the Example compounds were stably bound to STAT3 protein (see Figure 1 ).

[0501] Experimental Example 4: Thermal Shift Assay to Evaluate the Binding Effect of Example Compounds on STAT3 Protein

[0502] 1) Principle: Thermal shift assay (TSA) is a biophysical method based on the fact that the thermal stability of a target protein usually increases after binding to a ligand.

[0503] 2) Experimental Methods: Purified STAT3 protein was incubated with the example compounds (100 μM) or solvent (DMSO) at room temperature for 30 minutes and then refrigerated. The protein was then heated at various temperatures (39-55°C) for 3 minutes and cooled at room temperature for 3 minutes. All samples were centrifuged, and the supernatants were analyzed by Western blotting.

[0504] 3) The results showed that the example compounds significantly increased the accumulation of STAT3 in the temperature range of 39 to 55°C ( Figure 2 ), indicating that the compound can directly bind to STAT3 protein and increase the thermal stability of STAT3 protein.

[0505] Experimental Example 5: Immunoblotting to evaluate the inhibitory effect of the example compounds on intracellular STAT3 protein phosphorylation

[0506] MDA-MB-231 cells and MDA-MB-468 cells treated with the control group and different concentrations of the example compounds for 6 hours were collected, washed twice with pre-chilled PBS, and lysed on ice for 1 hour after adding an appropriate amount of RIPA lysis buffer (50mM Tris-HCl, 1mM EDTA, 1% Triton X-100, 150mM NaCl, 0.1% SDS, 1mM NaF, Na3VO4, protease inhibitors, pH 7.4). The cells were centrifuged at 4°C, 12,000 rpm for 20 minutes, and the supernatant was collected for protein quantification and boiled for denaturation. Equal amounts of protein were subjected to 10% SDS-PAGE electrophoresis. Primary antibodies were used, such as p-STAT3 (Tyr705) and STAT3-specific antibodies, and incubated overnight at 4°C. The corresponding HRP-labeled secondary antibodies were used for incubation at room temperature for 2 hours and washed. ECL chemiluminescent substrate reaction solution was added, the cells were developed in a gel imaging system, and the images were saved. β-actin was used as an internal reference.

[0507] The results showed that the example compounds could dose-dependently inhibit the phosphorylation of tyrosine 705 of STAT3 protein in MDA-MB-231 cells (see Figure 3 ), among which the example compounds have a more obvious inhibitory effect on p-STAT3 at the Y705 site.

[0508] Experimental Example 6: Immunoblotting to evaluate the inhibitory effect of the example compounds on STAT3 protein phosphorylation in MDA-MB-231 cells

[0509] MDA-MB-231 cells from the control group and those treated with different concentrations of the example compounds for 12 hours were collected, washed twice with pre-chilled PBS, and lysed on ice for 1 hour with an appropriate amount of RIPA lysis buffer (50mM Tris-HCl, 1mM EDTA, 1% Triton X-100, 150mM NaCl, 0.1% SDS, 1mM NaF, Na3VO4, protease inhibitors, pH 7.4). The cells were centrifuged at 4°C, 12,000 rpm for 20 minutes, and the supernatant was collected for protein quantification and denatured by boiling. Equal amounts of protein were subjected to 10% SDS-PAGE electrophoresis. Primary antibodies were used, such as p-STAT3 (Tyr705) and STAT3-specific antibodies, and incubated overnight at 4°C. The corresponding HRP-labeled secondary antibodies were incubated at room temperature for 2 hours and washed. ECL chemiluminescent substrate reaction solution was added, the cells were developed in a gel imaging system, and the images were saved. β-actin was used as an internal reference.

[0510] The results showed that the example compounds could dose-dependently inhibit the phosphorylation of tyrosine 705 of STAT3 protein in MDA-MB-231 cells (see Figure 4 ).

[0511] Experimental Example 7: Immunoblotting to evaluate the inhibitory effect of the example compounds on STAT3 protein phosphorylation in MDA-MB-468 cells

[0512] MDA-MB-468 cells were collected from the control group and treated with various concentrations of the example compounds for 6 and 12 hours, washed twice with pre-chilled PBS, and lysed on ice for 1 hour in RIPA lysis buffer (50mM Tris-HCl, 1mM EDTA, 1% Triton X-100, 150mM NaCl, 0.1% SDS, 1mM NaF, Na3VO4, protease inhibitors, pH 7.4). The cells were centrifuged at 4°C, 12,000 rpm for 20 minutes, and the supernatant was collected for protein quantification and denatured by boiling. Equal amounts of protein were subjected to 10% SDS-PAGE electrophoresis. Primary antibodies were used, including p-STAT3 (Tyr705) and STAT3-specific antibodies, and incubated overnight at 4°C. The cells were then incubated with the corresponding HRP-labeled secondary antibody at room temperature for 2 hours and washed. ECL chemiluminescent substrate reaction solution was added, the cells were developed in a gel imaging system, and the images were saved. β-actin was used as an internal reference.

[0513] The results showed that the example compounds could dose-dependently inhibit the phosphorylation of tyrosine 705 of STAT3 protein in MDA-MB-468 cells (see Figure 5 ).

Claims

1. The compound represented by the following formula I and its pharmaceutically acceptable salt Where: R1 is fluorine, chlorine, bromine, iodine, straight chain or branched C 1-6 Alkoxy, substituted and unsubstituted C 6-12 Phenoxy, substituted and unsubstituted N-heterocyclylamino, wherein the phenoloxy and N-heterocyclylamino groups can be unsubstituted or substituted by one or more groups selected from the following groups, these groups include straight chain or branched C 1-6 Alkyl, linear or branched C 1-6 Alkoxy, linear or branched amino, halogen, carboxyl, C 1-6 wherein the heterocyclic groups may be unsubstituted or substituted by one or more groups selected from the following groups, including C 1-6 Alkyl, C 1-6 Alkoxy, amino, halogen. R2 is hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy substituted C 1-6 Alkyl, alkyl-substituted C 1-6 Alkyl and C 3-6 Cycloalkyl, wherein the cycloalkyl may be unsubstituted or substituted by one or more groups selected from the following groups, these groups include C 1-6 Alkyl, C 1-6 Alkoxy, amino. R3 is hydrogen, C 1-6 Alkyl, substituted and unsubstituted heteroaryl, substituted and unsubstituted aryl, wherein the aryl or heteroaryl can be unsubstituted or substituted by one or more groups selected from the following groups, these groups include C 1-6 Alkyl, C 1-6 Alkoxy, amino, halogen, cyano, C 6-12 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, C 1-6 Ester group, hydroxyl group, C 1-6 Amide group, methylsulfone group. R4 is hydrogen, C 1-6 Alkyl, substituted and unsubstituted heteroaryl, substituted and unsubstituted aryl, wherein the aryl or heteroaryl can be unsubstituted or substituted by one or more groups selected from the following groups, these groups include C 1-6 Alkyl, C 1-6 Alkoxy, linear or branched substituted and unsubstituted amino, C 1-12 Cyclic amino, halogen, cyano, C 6-12 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, nitro, C 1-6 Ester group, hydroxyl group, C 1-6 Amide, C 1-6 Heteroaryl, C 4-12 Triazole, trifluoromethylphenyloxy, C 1-12 Sulfanyl, C 1-12 Sulfonamide, phenyl, morpholinyl, C 1-12 Alkynyl, C 1-6 Cycloalkoxy or C 1-12 Alkyleneoxy, hydrazine, C 1-12 Hydrazide group, C 5-12 Aromatic heterocycle.

2. The compound according to claim 1 and its pharmaceutically acceptable salt, characterized in that R1 is fluorine, chlorine, bromine, iodine, straight chain or branched C 1-4 Alkoxy, substituted and unsubstituted C 6-10 Phenoxy, substituted or unsubstituted N-heterocyclylamino, wherein the phenoloxy and N-heterocyclylamino groups can be unsubstituted or substituted by one or more groups selected from the following groups, these groups include straight chain or branched C 1-4 Alkyl, linear or branched C 1-4 Alkoxy, linear or branched amino, halogen, carboxyl, C 1-4 wherein the heterocyclic groups may be unsubstituted or substituted by one or more groups selected from the following groups, including C 1-4 Alkyl, C 1-4 Alkoxy, amino, halogen. R2 is hydrogen, C 1-4 Alkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, alkyl-substituted C 1-4 Alkyl and C 3-6 Cycloalkyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino. R3 is hydrogen, C 1-4 Alkyl, substituted and unsubstituted heteroaryl, substituted and unsubstituted aryl, wherein the aryl or aromatic heteroyl can be substituted or unsubstituted one or more groups selected from the following groups, these groups include C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, cyano, C 6-10 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, C 1-4 Ester group, hydroxyl group, C 1-4 Amide group, methyl sulfone group. R4 is hydrogen, C 1-4 Alkyl, substituted and unsubstituted heteroaryl, substituted and unsubstituted aryl, wherein the aryl or heteroaryl can be unsubstituted or substituted by one or more groups selected from the following groups, these groups include C 1-4 Alkyl, C 1-4 Alkoxy, linear or branched substituted and unsubstituted amino, C 1-10 Cyclic amino, halogen, cyano, C 6-10 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, nitro, C 1-4 Ester group, hydroxyl group, C 1-4 Amide, C 1-4 Heteroaryl, C 4-10 Triazole, trifluoromethylphenyloxy, C 1-10 Sulfanyl, C 1-10 Sulfonamide, phenyl, morpholinyl, C 1-10 Alkynyl, C 1-4 Cycloalkoxy or C 1-10 Alkyleneoxy, hydrazine, C 1-10 Hydrazide group, C 5-10 Aromatic heterocycle.

3. The compound according to claim 2 and its pharmaceutically acceptable salt, characterized in that R1 is fluorine, chlorine, bromine, iodine, straight chain or branched C 1-4 Alkoxy, substituted and unsubstituted phenoxy, substituted and unsubstituted naphthoxy, substituted or unsubstituted piperazinyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include straight chain or branched C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, carboxyl, C 1-4 ester group and hydroxyl group. R2 is hydrogen, C 1-4 Alkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, alkyl-substituted C 1-4 Alkyl and C 3-6 Cycloalkyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino. R3 is hydrogen, C 1-4 alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted naphthyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted pyrazinyl, substituted and unsubstituted pyridazinyl, substituted and unsubstituted benzimidazolyl, substituted and unsubstituted benzoxazolyl, substituted and unsubstituted benzothiazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, cyano, phenoxy, naphthoxy, trifluoromethyl, trifluoromethoxy, carboxyl, C 1-4 Ester group, hydroxyl group, C 1-4 Amide group, methyl sulfone group. R4 is hydrogen, C 1-4 alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted naphthyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted pyrazinyl, substituted and unsubstituted pyridazinyl, substituted and unsubstituted benzimidazolyl, substituted and unsubstituted benzoxazolyl, substituted and unsubstituted benzothiazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, linear or branched substituted and unsubstituted amino, C 1-10 Cyclic amino, halogen, cyano, C 6-10 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, nitro, C 1-4 Ester group, hydroxyl group, C 1-4 Amide, C 1-4 Heteroaryl, C 4-10 Triazole, trifluoromethylphenyloxy, C 1-10 Sulfanyl, C 1-10 Sulfonamide, phenyl, morpholinyl, C 1-10 Alkynyl, C 1-4 Cycloalkoxy or C 1-10 Alkyleneoxy, hydrazine, C 1-10 Hydrazide group, C 5-10 Aromatic heterocycle.

4. The compound according to claim 3 and a pharmaceutically acceptable salt thereof, characterized in that R1 is fluorine, chlorine, bromine, iodine, straight chain or branched C 1-4 Alkoxy, substituted and unsubstituted phenoxy, substituted and unsubstituted naphthoxy, substituted or unsubstituted piperazinyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include straight chain or branched C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, carboxyl, C 1-4 Ester group and hydroxyl group; R2 is hydrogen, C 1-4 Alkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, alkyl-substituted C 1-4 Alkyl and C 3-6 Cycloalkyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino. R3 is hydrogen, C 1-4 alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted benzimidazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, amino, fluorine, chlorine, bromine, iodine, cyano, phenoxy, naphthoxy, trifluoromethyl, trifluoromethoxy, carboxyl, C 1-4 Ester group, hydroxyl group, C 1-4 Amide group, methyl sulfone group. R4 is hydrogen, C 1-4 alkyl, substituted and unsubstituted phenyl, substituted and unsubstituted furyl, substituted and unsubstituted thienyl, substituted and unsubstituted pyrrolyl, substituted and unsubstituted oxazolyl, substituted and unsubstituted thiazolyl, substituted and unsubstituted imidazolyl, substituted and unsubstituted pyridyl, substituted and unsubstituted pyrimidinyl, substituted and unsubstituted benzimidazolyl, wherein the substituted or unsubstituted substituent is selected from one or more of the following groups, which include C 1-4 Alkyl, C 1-4 Alkoxy, linear or branched substituted and unsubstituted amino, C 1-10 Cyclic amino, halogen, cyano, C 6-10 Phenoxy, trifluoromethyl, trifluoromethoxy, carboxyl, nitro, C 1-4 Ester group, hydroxyl group, C 1-4 Amide, C 1-4 Heteroaryl, C 4-10 Triazole, trifluoromethylphenyloxy, C 1-10 Sulfanyl, C 1-10 Sulfonamide, phenyl, morpholinyl, C 1-10 Alkynyl, C 1-4 Cycloalkoxy or C 1-10 Alkyleneoxy, hydrazine, C 1-10 Hydrazide group, C 5-10 Aromatic heterocycle.

5. The compound according to claim 1-4 and a pharmaceutically acceptable salt thereof, wherein the compound is selected from: Compound 1: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine Compound 2: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine Compound 3: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 4: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 5: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 6: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine Compound 7: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1,2,4-triazolyl)phenyl)-1,3,4-thiadiazole-2-amine Compound 8: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazole-2-amine Compound 9: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine Compound 10: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine Compound 11: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 12: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 13: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(3-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine Compound 14: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-trifluoromethoxyphenyl)-1,3,4-thiadiazole-2-amine Compound 15: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(4-(N,N-dimethylamino)phenyl)-1,3,4-thiadiazole-2-amine Compound 16: N-(1-cyclopropyl-6-(4-ethylpiperazinyl)-2-(2-fluorophenyl)-5-benzimidazolyl)-5-(3-hydroxyphenyl)-1,3,4-thiadiazole-2-amine Compound 17: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 18: N-(1-cyclopropyl-6-fluoro-2-(4-fluorophenyl)-5-benzimidazolyl)-5-(4-(1-pyrrolidinyl)phenyl)-1,3,4-thiadiazole-2-amine Compound 19: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-cyanophenyl)-1,3,4-thiadiazole-2-amine Compound 20: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylphenyl)-1,3,4-thiadiazole-2-amine Compound 21: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-(3-trifluoromethylphenyl)oxyphenyl)-1,3,4-thiadiazole-2-amine Compound 22: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-methylthiophenyl)-1,3,4-thiadiazole-2-amine Compound 23: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(phenyl)phenyl)-1,3,4-thiadiazole-2-amine Compound 24: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4-dichlorophenyl)-1,3,4-thiadiazole-2-amine Compound 25: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(morpholinyl)phenyl)-1,3,4-thiadiazole-2-amine Compound 26: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-(methoxy)phenyl)-1,3,4-thiadiazole-2-amine Compound 27: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 28: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethyloxyphenyl)-1,3,4-thiadiazole-2-amine Compound 29: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 30: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 31: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 32: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazole-2-amine Compound 33: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine Compound 34: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 35: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine Compound 36: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-hydroxyphenyl)-1,3,4-thiadiazole-2-amine Compound 37: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-chlorophenyl)-1,3,4-thiadiazole-2-amine Compound 38: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-chlorophenyl)-1,3,4-thiadiazole-2-amine Compound 39: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-chlorophenyl)-1,3,4-thiadiazole-2-amine Compound 40: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-bromophenyl)-1,3,4-thiadiazole-2-amine Compound 41: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-bromophenyl)-1,3,4-thiadiazole-2-amine Compound 42: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 43: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-aminophenyl)-1,3,4-thiadiazole-2-amine Compound 44: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(2-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 45: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 46: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-propoxyphenyl)-1,3,4-thiadiazole-2-amine Compound 47: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-ethynylphenyl)-1,3,4-thiadiazole-2-amine Compound 48: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(3,4-ethylenedioxyphenyl)-1,3,4-thiadiazole-2-amine Compound 49: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-formamidophenyl)-1,3,4-thiadiazole-2-amine Compound 50: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-iodophenyl)-1,3,4-thiadiazole-2-amine Compound 51: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-iodophenyl)-1,3,4-thiadiazole-2-amine Compound 52: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-iodophenyl)-1,3,4-thiadiazole-2-amine Compound 53: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-hydroxyphenyl)-1,3,4-thiadiazole-2-amine Compound 54: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(2-ethoxyacylphenyl)-1,3,4-thiadiazole-2-amine Compound 55: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-ethoxyacylphenyl)-1,3,4-thiadiazole-2-amine Compound 56: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3-methanesulfonylaminophenyl)-1,3,4-thiadiazole-2-amine Compound 57: N-(1-cyclopropyl-2-(3-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 58: N-(1-cyclopropyl-2-(2-fluorophenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 59: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-isopropylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 60: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine Compound 61: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 62: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 63: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine Compound 64: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-(4-acetylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethoxyphenyl)-1,3,4-thiadiazole-2-amine Compound 65: N-(1-isopropyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 66: N-(1-pentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 67: N-(1-cyclopentyl-2-(4-fluorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 68: N-(1-cyclopropyl-2-phenyl-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 69: N-(1-cyclopropyl-2-(4-chlorophenyl)-6-(4-methylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 70: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 71: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine Compound 72: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 73: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine Compound 74: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine Compound 75: N-(1-cyclopropyl-2-(4-trifluoromethoxyphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine Compound 76: N-(1-cyclopropyl-2-(4-fluorophenyl)-6-fluoro-5-benzimidazolyl)-5-(3-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 77: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-nitrophenyl)-1,3,4-thiadiazole-2-amine Compound 78: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-cyanophenyl)-1,3,4-thiadiazole-2-amine Compound 79: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazole-2-amine Compound 80: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole-2-amine Compound 81: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole-2-amine Compound 82: N-(1-cyclopropyl-2-(4-trifluoromethylphenyl)-6-(4-ethylpiperazinyl)-5-benzimidazolyl)-5-phenyl-1,3,4-thiadiazole-2-amine 6. A pharmaceutical composition, characterized in that The pharmaceutical composition contains a pharmaceutically effective amount of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable carrier or excipient.

7. Use of the compound according to any one of claims 1 to 5 and a pharmaceutically acceptable salt thereof in the preparation of a drug for inhibiting STAT3 activity.

8. Use of the compound according to any one of claims 1 to 5 and a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing or treating cancer.

9. Use according to claim 8, characterized in that The cancer is selected from breast cancer, prostate cancer, ovarian cancer, liver cancer, gastric cancer, lung cancer, colon cancer, pancreatic cancer, esophageal cancer, leukemia, human brain glioma, and lymphoma.

10. A method for preparing the compound of claim 1 and a pharmaceutically acceptable salt thereof, comprising the steps of: a) monosubstitution; b) disubstitution; c) reduction; d) cyclization; e) formation of isothiocyanate; f) formation of thiosemicarbazide group; g) condensation; h) cyclization; It is characterized in that The steps include: 1) The compound of formula (1) undergoes a mono-substitution reaction with a primary amine to obtain a compound of formula (2); 2) the compound of formula (2) continues to undergo a disubstitution reaction with a secondary amine or a primary alcohol to obtain a compound of formula (3); 3) The compound of formula (2) or formula (3) is subjected to palladium carbon reduction-ammonium formate reduction and cyclization to obtain the compound of formula (4); 4) The compound of formula (4) reacts with thiocarbonyldiimidazole or thiocarbonyldiimidazole to form an isothiocyanate, which is then reacted with hydrazine hydrate to obtain a compound of formula (5); 5) The compound of formula (5) undergoes condensation reaction with aldehyde, and then undergoes cyclization in the presence of ferric chloride to obtain the compound of formula (I); Wherein R1, R2, R3, and R4 are defined as those in claims 1-4.

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