Thiadiazole and pyridazine parent nucleus compound and preparation method and pharmaceutical application thereof

Abstract

The invention belongs to the field of medicines, and relates to a compound containing a thiadiazole and pyridazine parent nucleus structure skeleton or pharmaceutically acceptable salt thereof, a preparation method thereof, a pharmaceutical composition containing the compound and medical application of the compound. The compound provided by the invention has the effect of specifically inhibiting GAC protein, and can be used for preparing drugs for treating diseases related to GAC protein functions, such as malignant tumors.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a class of compounds containing thiadiazole and pyridazine parent core structures or pharmaceutically acceptable salts thereof, a preparation method thereof, a pharmaceutical composition containing the compounds, and the use of the compounds in preparing drugs for treating diseases related to GAC function. Background Art

[0002] Glutamine is an important carbon and nitrogen source for cellular life activities, and its metabolite glutamate can participate in redox balance, tricarboxylic acid cycle and the synthesis of various non-essential amino acids. Glutaminase is the rate-limiting enzyme of glutamine metabolism, which is divided into glutaminase 1 (GLS1) distributed throughout the body and glutaminase 2 (GLS2) distributed in the liver. GLS1 gene alternative splicing is expressed as two isozymes - kidney-type glutaminase (KGA) present in the cytoplasm and glutaminase C (GAC) present in the mitochondria. Studies have shown that the expression of GAC in mitochondria is closely related to the occurrence and development of tumors, and in tumor cells, the GAC form seems to play a core role in the metabolic changes of many proliferating cells. In many cancers, GAC is more actively expressed than KGA, indicating that alternative splicing of GLS1 may play an important role in the higher glutamine decomposition flux speculated in tumors. Therefore, inhibiting GAC as a new method for treating cancer has received considerable attention. The N-terminal sequence of GAC consists of 1-550 amino acids, and the C-terminal sequence consists of 551-598 amino acids. The tetramer is the active form of GAC in which the enzyme catalyzes its activity, and controls whether the substrate glutamine can enter the active site through the switch of the gate ring structure. It has four substrate catalytic sites and two allosteric binding sites.

[0003] The existing GAC inhibitors are mainly divided into the following categories. The first category is glutamate analogs, DON can covalently bind to the active site Ser286. In preclinical studies, these three drugs showed strong inhibitory effects in both in vitro culture and in vivo mouse transplant tumor models. Some studies have also shown that DON has shown good effects in several clinical trials. However, glutamine analogs have serious off-target effects. Therefore, the clinical application of DON and its diazo analogs is limited by dose-limiting toxicity and a narrow therapeutic window. The second category is BPTES and its derivatives with a thiadiazole structure, which has a completely new skeleton. The X-ray crystal structure of the GAC-BPTES complex shows that BPTES occupies an allosteric binding position and effectively captures GAC in the form of an inactive tetramer. At present, CB-839 is the only GAC inhibitor that has been clinically studied. It is modified from BPTES, replacing one thiadiazole ring with pyridazine, and replacing the two terminal benzene rings with pyridine and trifluoromethoxy substituted benzene rings. The third category is benzophenanthridine ketone derivatives. This type of inhibitor is an allosteric modulator of recombinant GAC, and its binding site is located in the cavity of the dimer formed by two GAC monomers. Studies have shown that this type of inhibitor has no inhibitory effect on activated glutaminase protein. This shortcoming and hydrophobicity limit its further application and continued research in the anti-tumor field. The fourth type is thiazolidine-2,4-dione derivatives, which identified the initial hit compound Yeh-5 through high-throughput screening against KGA. This type of inhibitor inhibits GAC by having a new binding site.

[0004] Summary of the invention

[0005] The present invention provides a class of thiadiazole and pyridazine parent nucleus compounds or pharmaceutically acceptable salts thereof, a preparation method thereof, a pharmaceutical composition containing the class of compounds and the use of the class of compounds in the preparation of drugs for treating diseases related to the function of GAC protein.

[0006] The present invention designs and synthesizes a series of GAC protein inhibitors with salt bridge force and π-cation force substituents based on the newly discovered hot spots Arg317 and Lys320 composed of basic amino acid residues in GAC protein.

[0007] In order to solve the technical problem of the present invention, the present invention provides the following technical solutions:

[0008] The first aspect of the technical solution of the present invention is to provide a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof:

[0009]

[0010] in,

[0011] R1 is selected from H, F, Cl, Br, I, CF3;

[0012] R2 is selected from H, OCH3, C 1-6 alkyl;

[0013] R3 is selected from H;

[0014] Or R2 and R3 are connected together to form a C 3-6 Cycloalkyl;

[0015] Ar is selected from substituted or unsubstituted aromatic rings or aromatic heterocycles or phenylmethyl; the aromatic rings or aromatic heterocycles are selected from five-membered rings, six-membered rings or naphthalene rings; the substituents are selected from C 1-3 Alkyl, C 1-3 Alkoxy, halogen, halogen-substituted alkyl, halogen-substituted alkoxy, hydroxy, hydroxymethyl, cyano, phenyl, phenoxy, phenyl-substituted alkyl, carboxyl, carboxylate, amide, alkyl-substituted or halogen-substituted alkyl-substituted amide, phosphate, phosphate, boric acid, borate, tetrazolyl and alkylsulfonylcarbonyl; the aromatic heterocycle contains 1-2 heteroatoms selected from N, O and S;

[0016] Ring L is selected from substituents having the following structure:

[0017]

[0018] As a preferred embodiment, in the general formula (I), Ar is selected from unsubstituted or substituted methyl, hydroxyl, hydroxymethyl, methoxy, trifluoromethyl, trifluoromethoxy, F, Cl, Br, cyano, carboxyl, phosphate, borate, tetrazolyl,

[0019] An aromatic ring or aromatic heterocyclic group substituted with any one or more substituents; the aromatic ring group is selected from phenyl and naphthyl; the aromatic heterocyclic group is selected from pyridyl, pyrrolyl, furyl and thienyl.

[0020] As a preferred embodiment, the pharmaceutically acceptable salt is selected from organic acid salts or inorganic acid salts, including hydrochlorides, hydrobromides, sulfates, phosphates, acetates, citrates, malates, fumarates, tartrates or methanesulfonates.

[0021] As a preferred embodiment of the present invention, the compound of general formula (I) has the following structure:

[0022]

[0023]

[0024]

[0025]

[0026]

[0027] Another aspect of the technical solution of the present invention is to provide a method for synthesizing a compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, and the synthetic route is as follows:

[0028] Route (1):

[0029]

[0030] R=Cl or CF3

[0031] Wherein R1, R2, R3, Ar are defined as described in any one of claims 1-4.

[0032] Reagents and conditions: (a) N-tert-butoxycarbonyl substituted diamine, N,N-diisopropylethylamine, acetonitrile, 80°C; (b) ammonium formate, palladium / carbon, ethanol; (c) trifluoroacetic acid, dichloromethane; (d) 5-bromo-1,3,4-thiadiazol-2-amine, N,N-diisopropylethylamine, isopropanol, 45°C; (e) unsubstituted phenylpropionic acid or substituted aromatic acetic acid, propylphosphonic anhydride, N,N-diisopropylethylamine, N,N-dimethylformamide.

[0033] Another aspect of the technical solution of the present invention is to provide a pharmaceutical composition, which comprises a compound represented by general formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0034] Another aspect of the technical solution of the present invention is to provide the use of the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating diseases related to the function of the GAC protein.

[0035] Beneficial technical effects

[0036] Examples 1-49 in the technical solution of the present invention all have significant inhibition on GAC protein, thereby blocking the occurrence and development of tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 Tumor volume-time curves of oral administration of CB839, Example 22 and Example 35 in the mouse A549 xenograft tumor model

[0038] Figure 2 The weight-time curves of CB839, Example 22 and Example 35 after oral administration in the mouse A549 xenograft tumor model DETAILED DESCRIPTION

[0039] Example 1

[0040] (R)-1-phenyl-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)cyclopropane-1-carboxamide

[0041]

[0042] (1) Preparation of tert-butyl (R)-(1-(6-chloropyridazin-3-yl)pyrrolidin-3-yl)carbamate

[0043]

[0044] 3,6-dichloropyridazine (1.00 g, 6.71 mmol), (R)-tert-butyl pyrrolidin-3-ylcarbamate (1.50 g, 8.06 mmol) and N,N-diisopropylethylamine (1.04 g, 8.06 mmol) were added to the bottle in sequence, dissolved in 10 mL of acetonitrile, and heated to 80 ° C for 5 h. The reaction was cooled to room temperature, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic layer was washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentration and column chromatography were performed, and the eluent was petroleum ether: ethyl acetate = 1:1 to obtain 1.62 g of white solid, with a yield of 81.2% and a melting point of 219-220 ° C.

[0045] 1 H NMR(400MHz,Chloroform-d)δ7.18(d,J=9.4Hz,1H),6.62(d,J=9.4Hz,1H),4.84(s,1H),4.37(s,1H),3.77(dd,J=10.9,6 .1Hz,1H),3.70–3.55(m,2H),3.42(dd,J=10.9,4.3Hz,1H),2.31(dq,J=13.7,7.0Hz,1H),2.08–1.98(m,1H),1.45(s,9H).

[0046] MS (ESI) m / z 299.00 (M+H) + .

[0047] (2) Preparation of tert-butyl (R)-(1-(pyridazin-3-yl)pyrrolidin-3-yl)carbamate

[0048]

[0049] Dissolve (R)-tert-butyl (1-(6-chloropyridazin-3-yl)pyrrolidin-3-yl)carbamate (1.00 g, 3.35 mmol), ammonium formate (2.10 g, 33.50 mmol) and palladium / carbon (100 mg, 10%) in 10 mL of ethanol and react at room temperature for 3 h. Remove the insoluble solid by filtration through diatomaceous earth, and distill the filtrate under reduced pressure to obtain 879 mg of a white solid with a yield of 99.4% and a melting point of 133-134°C.

[0050] 1 H NMR(400MHz,Chloroform-d)δ8.55(s,1H),7.237–7.15f(m,1H),6.667–6.60(m,1H),4.80(s,1H),4.39(s,1H), 3.87–3.75(m,1H),3.66(s,2H),3.49–3.40(m,1H),2.35–2.28(m,1H),2.07–1.99(m,1H),1.46(d,J=4.9Hz,9H).

[0051] MS (ESI) m / z 265.09 (M+H) + .

[0052] (3)(R)-N 2 Preparation of -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine

[0053]

[0054] Dissolve (R)-(1-(pyridazine-3-yl)pyrrolidin-3-yl)carbamic acid tert-butyl ester (1.00g, 3.79mmol) in 10mL dichloromethane, add 5mL trifluoroacetic acid dropwise in an ice-water bath, transfer to room temperature for reaction for 1h after addition, remove the solvent by distillation under reduced pressure, dissolve in isopropanol solution, add 5-bromo-1,3,4-thiadiazole-2-amine (818mg, 4.55mmol) and N,N-diisopropylethylamine (588mg, 4.55mmol), mix well, transfer to 45℃ and heat for reaction for 4h, remove the solvent by distillation under reduced pressure, add water to quench the reaction, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Concentrate, separate by column chromatography, and the eluent is dichloromethane:methanol=10:1. Obtain 683mg of off-white solid, yield 68.5%, melting point 230-232℃.

[0055] 1H NMR (400MHz, DMSO-d6) δ8.47(d,J=4.4Hz,1H),7.36–7.30(m,1H),7.11(d,J=6.0Hz,1H),6.86(d,J=9.1Hz,1H),6.33(s ,2H),4.27(s,1H),3.74–3.66(m,1H),3.59–3.50(m,2H),3.46(d,J=10.1Hz,1H),2.30–2.19(m,1H),2.06–1.96(m,1H).

[0056] MS (ESI) m / z 263.99 (M+H) + .

[0057] (4) Preparation of the title compound

[0058] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 1-phenylcyclopropane-1-acid (31 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 36 mg of white solid powder with a yield of 55.0% and a melting point of 125-126°C. 1 H NMR (400MHz, DMSO-d6) δ8.47(d,J=4.5Hz,1H),7.63(d,J=6.1Hz,1H),7.39–7.25(m,5H),6.87(d,J=9.2Hz,1H) ,4.37(s,1H),3.78–3.70(m,1H),3.62–3.41(m,3H),2.34–2.23(m,1H),2.06(s,1H),1.50(s,2H),1.17(s,2H).

[0059] HRMS calcd.For C20H22N7OS(M+H)+408.1601,found 408.1596.

[0060] Example 2

[0061] (R)-3-phenyl-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)propanamide

[0062]

[0063] (1) Preparation of the title compound

[0064] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), phenylpropionic acid (29 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 47 mg of white solid powder with a yield of 74.6% and a melting point of 225-226°C.

[0065] 1 H NMR(400MHz,DMSO-d6)δ11.97(s,1H),8.50(d,J=4.4Hz,1H),7.67(s,1H),7 .37(dd,J=9.0,4.2Hz,1H),7.29(d,J=7.4Hz,2H),7.25–7.19(m,3H),6.93( d,J=9.1Hz,1H),4.40(s,1H),3.83–3.74(m,1H),3.69–3.49(m,3H),2.90(t ,J=7.7Hz,2H),2.71(t,J=7.8Hz,2H),2.41–2.25(m,1H),2.20–2.03(m,1H).

[0066] HRMS calcd.For C19H22N7OS(M+H)+396.1601,found 396.1598.

[0067] Example 3

[0068] (R)-2-(3-bromophenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0069]

[0070] (1) Preparation of the title compound

[0071] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-bromophenylacetic acid (41 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 44 mg of white solid powder with a yield of 60.5% and a melting point of 213-215°C.

[0072] 1 H NMR (400MHz, DMSO-d6) δ12.21(s,1H),8.47(d,J=3.5Hz,1H),7.65(d,J=5.9Hz,1H),7.53(s,1H),7.46(s,1H),7.30(s ,3H),6.87(d,J=9.2Hz,1H),4.38(s,1H),3.74(s,3H),3.61–3.46(m,3H),2.34–2.22(m,1H),2.14–2.02(m,1H).HRMS calcd.ForC18H19BrN7OS(M+H)+460.0550,found 460.0550.

[0073] Example 4

[0074] (R)-2-(3-Fluorophenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0075]

[0076] (1) Preparation of the title compound

[0077] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-fluorophenylacetic acid (29 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 36 mg of white solid powder with a yield of 56.8% and a melting point of 231-232°C.

[0078] 1 H NMR (400MHz, DMSO-d6) δ12.24(s,1H),8.50(s,1H),7.68(d,J=6.0Hz,1H),7.45–7.32(m,2H),7.19–7.07(m,3H),6. 90(d,J=9.1Hz,1H),4.40(s,1H),3.81–3.73(m,3H),3.62–3.50(m,3H),2.37–2.25(m,1H),2.14–2.04(m,1H).HRMS calcd.ForC18H19FN7OS(M+H)+400.1350,found 400.1345.

[0079] Example 5

[0080] (R)-2-(3-Trifluoromethylphenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0081]

[0082] (1) Preparation of the title compound

[0083] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-trifluoromethylphenylacetic acid (39 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 44 mg of white solid powder with a yield of 61.5% and a melting point of 244-245°C. 1 H NMR (400MHz, DMSO-d6) δ12.25(s,1H),8.47(s,1H),7.72–7.54(m,5H),7.37–7.27(m,1H),6.87(d,J=9.1Hz,1H),4. 38(s,1H),3.86(s,2H),3.75(dd,J=11.0,5.8Hz,1H),3.60–3.46(m,3H),2.34–2.24(m,1H),2.13–1.99(m,1H).HRMS calcd.For C19H19F3N7OS(M+H)+450.1318,found 450.1315.

[0084] Example 6

[0085] (R)-2-(3-trifluoromethoxyphenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)

[0086] Acetamide

[0087]

[0088] (1) Preparation of the title compound

[0089] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-trifluoromethoxyphenylacetic acid (42 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 43 mg of white solid powder with a yield of 57.3% and a melting point of 214-216°C.

[0090] 1 H NMR (400MHz, DMSO-d6) δ12.23(s,1H),8.47(s,1H),7.66(d,J=6.0Hz,1H),7.46(t,J=7.9Hz,1H),7.33(d,J=7.8Hz,3H),7.26(d,J=8.5Hz,1 H),6.86(d,J=9.1Hz,1H),4.38(d,J=7.2Hz,1H),3.81(s,2H),3.77–3.71(m,1H),3.58–3.46(m,3H),2.34–2.23(m,1H),2.12–2.01(m,1H).

[0091] HRMS calcd.For C19H19F3N7O2S(M+H)+466.1268,found 466.1262.

[0092] Example 7

[0093] (R)-2-(3-Hydroxyphenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0094]

[0095] (1) Preparation of the title compound

[0096] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-hydroxyphenylacetic acid (29 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 42 mg of white solid powder with a yield of 66.2% and a melting point of 179-180°C.

[0097] 1 H NMR(400MHz,DMSO-d6)δ12.16(s,1H),9.36(s,1H),8.48(d,J=4.3Hz,1H),7 .64(d,J=6.1Hz,1H),7.37–7.31(m,1H),7.10(t,J=7.8Hz,1H),6.87(d,J=9. 1Hz,1H),6.71(s,2H),6.64(d,J=8.1Hz,1H),4.38(s,1H),3.75(d,J=11.3Hz ,1H),3.60(s,2H),3.58–3.46(m,3H),2.38–2.24(m,1H),2.13–1.97(m,1H).

[0098] HRMS calcd.For C18H20N7O2S(M+H)+398.1394,found 398.1390.

[0099] Example 8

[0100] (R)-2-(4-Hydroxymethylphenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0101]

[0102] (1) Preparation of the title compound

[0103] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 4-hydroxymethylphenylacetic acid (29 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise. The reaction solution was cooled to 0°C, and 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise. The reaction was moved to room temperature for 4 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 28 mg of white solid powder with a yield of 42.3% and a melting point of 224-225°C. 1 H NMR(400MHz,DMSO-d6)δ12.20(s,1H),8.50(d,J=4.4Hz,1H),7.66(s,1H),7 .35(dd,J=9.3,4.5Hz,1H),7.27(s,4H),6.89(d,J=8.9Hz,1H),5.16(d,J=5 .8Hz,1H),4.48(d,J=5.5Hz,2H),4.40(s,1H),3.77(dd,J=11.2,5.7Hz,1H) ,3.71(s,2H),3.63–3.47(m,4H),2.38–2.25(m,1H),2.10(d,J=10.6Hz,1H).

[0104] HRMS calcd.For C19H22N7O2S(M+H)+411.1550,found 411.1546.

[0105] Example 9

[0106] (R)-2-(3-methoxyphenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0107]

[0108] (1) Preparation of the title compound

[0109] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-methoxyphenylacetic acid (29 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 44 mg of white solid powder with a yield of 67.3% and a melting point of 178-180°C. 1 H NMR (400MHz, DMSO-d6) δ12.17(s,1H),8.47(s,1H),7.64(d,J=6.0Hz,1H),7.36–7.29(m,1H),7.23(t,J=7.8Hz,1H),6.92–6 .80(m,4H),4.38(s,1H),3.73(s,3H),3.68(s,2H),3.58–3.46(m,3H),2.32–2.23(m,1H),2.17–1.99(m,1H).HRMScalcd.For C19H22N7O2S(M+H)+411.1550,found 411.1546.

[0110] Example 10

[0111] (R)-2-(3-cyanophenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0112]

[0113] (1) Preparation of the title compound

[0114] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-cyanophenylacetic acid (31 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 36 mg of white solid powder with a yield of 55.6% and a melting point of 208-209°C.

[0115] 1 H NMR (400MHz, DMSO-d6) δ12.24(s,1H),8.47(d,J=4.4Hz,1H),7.75(d,J=9.0Hz,2H),7.65(t,J=6.8Hz,2H),7.54(t,J=7.7Hz,1H),7.35–7.30(m ,1H),6.87(d,J=9.2Hz,1H),4.38(s,1H),3.83(s,2H),3.75(dd,J=11. 0,5.9Hz,1H),3.60–3.47(m,3H),2.35–2.24(m,1H),2.14–2.00(m,1H).

[0116] HRMS calcd.For C19H19N8OS(M+H)+407.1397,found 407.1393.

[0117] Embodiment 11

[0118] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)aminoethylbenzamide

[0119]

[0120] (1) Preparation of 3-(2-(benzyloxy)-2-oxoethyl)benzoic acid

[0121]

[0122] 3-Carboxyphenylacetic acid (1 g, 5.55 mmol) was dissolved in 10 mL of anhydrous dichloromethane, benzyl alcohol (2.99 g, 27.75 mmol) was added, and 2 drops of thionyl chloride were added dropwise. The mixture was reacted at room temperature for 3 h in a dry environment until the insoluble solid disappeared. The solvent was removed by distillation under reduced pressure, and a saturated aqueous solution of ammonium chloride was added to quench the reaction. The mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was petroleum ether:ethyl acetate = 4:1 to obtain 1.43 g of white crystals with a yield of 95.6% and a melting point of 74-75°C.

[0123] 1 H NMR (400MHz, DMSO-d6) δ7.89 (s, 1H), 7.84 (d, J = 7.7Hz, 1H), 7.53 (d, J = 7.6Hz, 1H), 7. 45(t,J=7.6Hz,1H),7.34(s,5H),5.12(s,2H),3.85(s,2H).MS(ESI)m / z270.97(M+H) + .

[0124] (2) Preparation of 2-(3-carbamoylphenyl)benzyl acetate

[0125]

[0126] 3-(2-(Benzyloxy)-2-oxoethyl)benzoic acid (1 g, 3.72 mmol) was dissolved in 10 mL of N,N-dimethylformamide, and an isopropanol solution of ammonia (2.22 mL, 2 M) and N,N-diisopropylethylamine (1.92 g, 14.88 mmol) were added dropwise. Finally, HATU condensation agent (1.70 g, 4.46 mmol) was added. The reaction was allowed to react at room temperature for 3 h, and water was added to quench the reaction. The reaction was extracted with ethyl acetate three times, and the mixture was washed with saturated sodium chloride solution twice. The organic phase was concentrated and separated by column chromatography. The eluent was petroleum ether:ethyl acetate = 1:1 to obtain 670 mg of a light yellow oil with a yield of 67.0%.

[0127] 1 H NMR(700MHz,DMSO-d6)δ7.95(s,2H),7.79(s,1H),7.76(d,J=7.6Hz,1H),7.43(d, J=7.7Hz,1H),7.40(d,J=7.6Hz,1H),7.37–7.30(m,5H),5.12(s,2H),3.80(s,2H).

[0128] MS (ESI) m / z 270.02 (M+H) + .

[0129] (3) Preparation of 2-(3-carbamoylphenyl)acetic acid

[0130]

[0131] Benzyl 2-(3-carbamoylphenyl)acetate (1 g, 3.72 mmol) was dissolved in 10 mL of methanol, Raney Ni was added, hydrogen was introduced, and the reaction was carried out at room temperature for 2 h. After the catalyst was filtered off, the solvent was removed by distillation under reduced pressure to obtain 664 mg of white crystals, with a yield of 99.8% and a melting point of 164-165°C.

[0132] 1 H NMR(500MHz,DMSO-d6)δ7.96(s,1H),7.76(s,2H),7.36(m,2H),3.60(s,2H).MS(ESI)m / z 179.95(M+H) + .

[0133] (4) Preparation of the title compound

[0134] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-carbamoylphenyl)acetic acid (34 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise. The reaction solution was cooled to 0°C, and 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise. The reaction was moved to room temperature for 4 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 50 mg of a white solid powder with a yield of 74.3% and a melting point of 208-209°C.

[0135] 1 H NMR (400MHz, DMSO-d6) δ12.27(s,1H),8.50(d,J=4.4Hz,1H),7.98(s,1H),7.83(s,1H),7.78(d,J=7.5Hz,1H),7.68(d,J=5.5Hz,1H),7.49–7.33(m ,4H),6.92(d,J=9.2Hz,1H),4.40(s,1H),3.79(s,2H),3.75(d,J=1.6Hz, 1H),3.58(s,2H),3.52(d,J=11.2Hz,1H),2.39–2.23(m,1H),2.09(s,1H).

[0136] HRMS calcd.For C19H21N8O2S(M+H)+425.1503,found 425.1499.

[0137] Example 12

[0138] (R)-N-methyl-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzamide

[0139]

[0140] (1) Preparation of 2-(3-(methylcarbamoyl)phenyl)benzyl acetate

[0141]

[0142] 3-(2-(Benzyloxy)-2-oxoethyl)benzoic acid (1 g, 3.72 mmol) and methylamine hydrochloride (302 mg, 4.46 mmol) were dissolved in 10 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (1.92 g, 14.88 mmol) were added, and finally HATU condensation agent (1.70 g, 4.46 mmol) was added. The reaction was allowed to react at room temperature for 3 h, and water was added to quench the reaction. The reaction was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was petroleum ether:ethyl acetate = 1:1 to obtain 589 mg of light yellow oil with a yield of 55.9%.

[0143] 1 H NMR (300MHz, Chloroform-d) δ7.67 (dt, J = 8.4, 2.1Hz, 2H), 7.46–7.24 (m, 7H), 5.14 (s, 2H), 3.71 (s, 2H), 2.99 (s, 3H).

[0144] MS (ESI) m / z 284.03 (M+H) + .

[0145] (2) Preparation of 2-(3-(methylcarbamoyl)phenyl)acetic acid

[0146]

[0147] Dissolve 2-(3-(methylcarbamoyl)phenyl)benzyl acetate (1 g, 3.53 mmol) in 10 mL of methanol, add Raney Ni, introduce hydrogen, and react at room temperature for 2 h. After filtering the catalyst, remove the solvent by distillation under reduced pressure to obtain 674 mg of white crystals with a yield of 98.9% and a melting point of 110-111°C

[0148] 1 H NMR (300MHz, DMSO-d6) δ8.43(s,1H),7.71(s,2H),7.38(d,J=4.7Hz,2H),3.65–3.57(m,2H),2.77(d,J=3.9Hz,3H).

[0149] MS (ESI) m / z 194.05 (M+H) + .

[0150] (3) Preparation of the title compound

[0151] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-2-(3-(methylcarbamoyl)phenyl)acetic acid (37 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 29 mg of white solid powder with a yield of 67.3% and a melting point of 266-267°C.

[0152] 1 H NMR(300MHz,DMSO-d6)δ12.28(s,1H),8.49–8.40(m,2H),7.77(s,1H),7.74–7.63(m,2H),7.49–7.38(m,2H),7.40–7.27(m,1H),6.8 6(d,J=9.4Hz,1H),4.38(s,1H),3.81–3.69(m,3H),3.62–3.49(m,3H),2.76(d,J=4.5Hz,3H),2.32–2.22(m,1H),2.12–2.03(m,1H).

[0153] HRMS calcd.For C20H23N8O2S(M+H)+439.1659,found 439.1658.

[0154] Embodiment 13

[0155] (R)-N,N-dimethyl-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazole-2-

[0156] (amino)ethyl)benzamide

[0157]

[0158] (1) Preparation of 2-(3-(dimethylcarbamoyl)phenyl)benzyl acetate

[0159]

[0160] 3-(2-(Benzyloxy)-2-oxoethyl)benzoic acid (1 g, 3.72 mmol) and dimethylamine hydrochloride (364 mg, 4.46 mmol) were dissolved in 10 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (1.92 g, 14.88 mmol) were added, and finally HATU condensation agent (1.70 g, 4.46 mmol) was added. The reaction was allowed to react at room temperature for 3 h, and water was added to quench the reaction. The reaction was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was petroleum ether:ethyl acetate = 1:1 to obtain 693 mg of light yellow oil with a yield of 62.7%.

[0161] 1 H NMR (400MHz, DMSO-d6) δ7.40–7.33(m,6H),7.31–7.26(m,2H),5.12(s,2H),3.80(s,2H),2.97(s,3H),2.86(s,3H).

[0162] MS (ESI) m / z 298.05 (M+H) + .

[0163] (2) Preparation of 2-(3-(dimethylcarbamoyl)phenyl)acetic acid

[0164]

[0165] Benzyl 2-(3-(dimethylcarbamoyl)phenyl)acetate (1 g, 3.37 mmol) was dissolved in 10 mL of methanol, Raney Ni was added, hydrogen was introduced, and the reaction was carried out at room temperature for 2 h. After the catalyst was filtered off, the solvent was removed by distillation under reduced pressure to obtain 697 mg of a light yellow oil with a yield of 99.9%.

[0166] 1 H NMR (400MHz, DMSO-d6) δ7.41–7.30(m,3H),7.27(d,J=9.1Hz,1H),3.62(s,2H),2.97(s,3H),2.90(s,3H).

[0167] MS (ESI) m / z 207.96 (M+H) + .

[0168] (3) Preparation of the title compound

[0169] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-2-(3-(dimethylcarbamoyl)phenyl)acetic acid (39 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphoric anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 46 mg of white solid powder with a yield of 63.5% and a melting point of 148-150°C.

[0170] 1 H NMR (400MHz, DMSO-d6) δ12.25(s,1H),8.50(d,J=4.4Hz,1H),7.67(d,J=6.0Hz,1H),7.45–7.34(m,4H),7.31(d,J=6.8Hz,1H),6.92 (d,J=9.2Hz,1H),4.40(s,1H),3.80–3.74(m,3H),3.64–3.48(m,3H),2.96(d,J=29.8Hz,6H),2.35–2.25(m,1H),2.15–2.05(m,1H).

[0171] HRMS calcd.For C21H25N8O2S(M+H)+453.1816,found 453.1810.

[0172] Embodiment 14

[0173] (R)-N-(2-fluoroethyl)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzamide

[0174]

[0175] (1) Preparation of 2-(3-((2-fluoroethyl)carbamoyl)phenyl)benzyl acetate

[0176]

[0177] 3-(2-(Benzyloxy)-2-oxoethyl)benzoic acid (1 g, 3.72 mmol) and 2-fluoroacetic acid hydrochloride (444 mg, 4.46 mmol) were dissolved in 10 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (1.92 g, 14.88 mmol) were added, and finally HATU condensation agent (1.70 g, 4.46 mmol) was added. The reaction was allowed to react at room temperature for 3 h, and water was added to quench the reaction. The reaction was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was petroleum ether:ethyl acetate = 1:1 to obtain 643 mg of light yellow oil with a yield of 54.8%.

[0178] 1 H NMR(400MHz,Chloroform-d)δ7.70(s,2H),7.44(s,2H),7.34(s,5H),6.44(s,1H),5.15(d,J=5.0Hz ,2H),4.67(d,J=5.4Hz,1H),4.55(d,J=4.9Hz,1H),3.82(s,1H),3.73(d,J=5.1Hz,2H),2.99(s,1H).

[0179] MS (ESI) m / z 315.95 (M+H) + .

[0180] (2) Preparation of 2-(3-((2-fluoroethyl)carbamoyl)phenyl)acetic acid

[0181]

[0182] Benzyl 2-(3-((2-fluoroethyl)carbamoyl)phenyl)acetate (1 g, 3.17 mmol) was dissolved in 10 mL of methanol, Raney Ni was added, hydrogen was introduced, and the reaction was carried out at room temperature for 2 h. After the catalyst was filtered off, the solvent was removed by distillation under reduced pressure to obtain 711 mg of a white solid with a yield of 99.7% and a melting point of 116-118°C.

[0183] 1 H NMR (400MHz, DMSO-d6) δ8.67(s,1H),7.74(s,2H),7.40(s,2H),4.58(d,J=5.3Hz,1H),4.47(t,J=4.8Hz,1H),3.55(dd,J=26.5,5.5Hz,4H).

[0184] MS (ESI) m / z 226.01 (M+H) + .

[0185] (3) Preparation of the title compound

[0186] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-2-(3-((2-fluoroethyl)carbamoyl)phenyl)acetic acid (43 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 52 mg of white solid powder with a yield of 68.7% and a melting point of 161-162°C.

[0187] 1 H NMR (400MHz, DMSO-d6) δ12.27(s,1H),8.71(s,1H),8.49(d,J=4.6Hz,1H),7.82(s,1H), 7.77(d,J=7.4Hz,1H),7.67(d,J=5.8Hz,1H),7.51–7.43(m,2H),7.35(dd,J=9.4,4.5Hz, 1H),6.89(d,J=9.3Hz,1H),4.61(t,J=5.0Hz,1H),4.49(t,J=5.0Hz,1H),4.40(s,1H),3 .81(s,2H),3.78–3.73(m,1H),3.65–3.48(m,5H),2.37–2.26(m,1H),2.16–2.04(m,1H).

[0188] HRMS calcd.For C21H24FN8O2S(M+H)+471.1721,found 471.1720.

[0189] Embodiment 15

[0190] (R)-N-(3,3-difluorocyclobutyl)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-

[0191] thiadiazole-2-yl)amino)ethyl)benzamide

[0192]

[0193] (1) Preparation of 2-(3-((3,3-difluorocyclobutyl)carbamoyl)phenyl)benzyl acetate

[0194]

[0195] 3-(2-(Benzyloxy)-2-oxoethyl)benzoic acid (1 g, 3.72 mmol) and 3,3-difluorocyclobutane-1-amine (478 mg, 4.46 mmol) were dissolved in 10 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (1.92 g, 14.88 mmol) were added, and finally HATU condensation agent (1.70 g, 4.46 mmol) was added. The reaction was reacted at room temperature for 3 h, and water was added to quench the reaction. The reaction was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated. The reaction was separated by column chromatography with petroleum ether: ethyl acetate = 1:1 as the eluent to obtain 753 mg of a white solid with a yield of 56.4% and a melting point of 125-126°C.

[0196] 1 H NMR (400MHz, Chloroform-d) δ7.65 (s, 2H), 7.41 (d, J = 7.9Hz, 2H), 7.37–7.28 (m, 5H), 6. 29(s,1H),5.13(s,2H),4.44(s,1H),3.70(s,2H),3.08(s,2H),2.57(d,J=14.4Hz,2H).

[0197] MS (ESI) m / z 360.05 (M+H) + .

[0198] (2) Preparation of 2-((3-(3,3-difluorocyclobutyl)carbamoyl)phenyl)acetic acid

[0199]

[0200] Benzyl 2-(3-((3,3-difluorocyclobutyl)carbamoyl)phenyl)acetate (1 g, 2.78 mmol) was dissolved in 10 mL of methanol, Raney Ni was added, hydrogen was introduced, and the reaction was carried out at room temperature for 2 h. After the catalyst was filtered off, the solvent was removed by distillation under reduced pressure to obtain 743 mg of a white solid with a yield of 99.3% and a melting point of 158-160°C.

[0201] 1H NMR(400MHz,DMSO-d6)δ8.81(d,J=6.7Hz,1H),7.73(s,2H),7.43(s,2H),4.27(s,1H),3 .64(s,1H),3.17(s,1H),2.94(s,2H),2.75(d,J=15.5Hz,2H).MS(ESI)m / z269.98(M+H) + .

[0202] (3) Preparation of the title compound

[0203] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-((3,3-difluorocyclobutyl)carbamoyl)phenyl)acetic acid (51 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, the reaction solution was moved to room temperature for 4 h, after which the reaction was quenched with water, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, the organic phase was concentrated, and separated by column chromatography with the eluent being dichloromethane:methanol = 10:1 to give 42 mg of a white solid powder with a yield of 51.2% and a melting point of 161-162°C.

[0204] 1 H NMR(400MHz,DMSO-d6)δ12.25(s,1H),8.82(s,1H),8.48(s,1H),7.78(s,1H),7.78–7.71(m,1H),7.65(s,1H),7.50–7.40(m,2H),7.39–7.28(m ,1H),6.91–6.84(m,1H),4.38(s,1H),4.27(s,1H),3.86–3.70(m,3H), 3.61–3.46(m,3H),2.94(s,2H),2.75(s,2H),2.29(s,1H),2.07(s,1H).

[0205] HRMS calcd.For C23H25F2N8O2S(M+H)+515.1784,found 515.1788.

[0206] Example 16

[0207] (R)-N-(4,4-difluorocyclohexyl)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-

[0208] thiadiazole-2-yl)amino)ethyl)benzamide

[0209]

[0210] (1) Preparation of 2-(3-((4,4-difluorocyclohexyl)carbamoyl)phenyl)benzyl acetate

[0211]

[0212] 3-(2-(Benzyloxy)-2-oxoethyl)benzoic acid (1 g, 3.72 mmol) and 4,4-difluorocyclohexane-1-amine hydrochloride (765 mg, 4.46 mmol) were dissolved in 10 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (1.92 g, 14.88 mmol) were added, and finally HATU condensation agent (1.70 g, 4.46 mmol) was added. The reaction was reacted at room temperature for 3 h, and water was added to quench the reaction. The reaction was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated. The reaction was separated by column chromatography with petroleum ether: ethyl acetate = 1:1 as the eluent to obtain 1.25 g of a white solid with a yield of 62.1% and a melting point of 132-133°C.

[0213] 1 H NMR(400MHz,Chloroform-d)δ7.65(s,2H),7.47–7.29(m,7H),5.94(s,1H),5.14(s,2H),4 .10(s,1H),3.72(s,2H),2.13(s,4H),1.94(m,2H),1.62(m,2H).MS(ESI)m / z387.86(M+H) + .

[0214] (2) Preparation of 2-(3-((4,4-difluorocyclohexyl)carbamoyl)phenyl)acetic acid

[0215]

[0216] Benzyl 2-(3-((4,4-difluorocyclohexyl)carbamoyl)phenyl)acetate (1 g, 2.58 mmol) was dissolved in 10 mL of methanol, Raney Ni was added, hydrogen was introduced, and the reaction was carried out at room temperature for 2 h. After the catalyst was filtered off, the solvent was removed by distillation under reduced pressure to obtain 764 mg of a white solid with a yield of 99.6% and a melting point of 158-160°C.

[0217] 1H NMR (400MHz, DMSO-d6) δ7.73(s,2H),7.40(s,2H),3.99(s,1H),3.63(s,2H),2.04(s,3H),1.87(m,3H),1.64(m,2H).

[0218] MS (ESI) m / z 298.04 (M+H) + .

[0219] (3) Preparation of the title compound

[0220] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-((4,4-difluorocyclohexyl)carbamoyl)phenyl)acetic acid (56 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, the reaction solution was moved to room temperature for 4 h, after which the reaction was quenched with water, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, the organic phase was concentrated, and separated by column chromatography with the eluent being dichloromethane:methanol = 10:1 to obtain 51 mg of a white solid powder with a yield of 58.6% and a melting point of 182-183°C.

[0221] 1 H NMR (400MHz, DMSO-d6) δ12.24(s,1H),8.47(d,J=4.4Hz,1H),8.29(d,J=7.7Hz,1H),7.77(s,1H ),7.73(d,J=7.2Hz,1H),7.64(d,J=6.0Hz,1H),7.42(dt,J=14.8,7.4Hz,2H),7.32(dd,J=9.2, 4.4Hz,1H),6.86(d,J=9.2Hz,1H),4.38(s,1H),3.99(s,1H),3.78(s,2H),3.77–3.70(m,1H),3 .60–3.46(m,3H),2.34–2.23(m,1H),2.12–1.97(m,4H),1.92–1.81(m,3H),1.72–1.58(m,2H).

[0222] HRMS calcd.For C25H29F2N8O2S(M+H)+543.2097,found 543.2096.

[0223] Embodiment 17

[0224] (R)-methyl 3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoate

[0225]

[0226] (1) Preparation of the title compound

[0227] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-(methoxycarbonyl)phenyl)acetic acid (37 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 40 mg of white solid powder with a yield of 56.6% and a melting point of 227-228°C.

[0228] 1 H NMR (400MHz, DMSO-d6) δ8.48 (s, 1H), 7.94 (s, 1H), 7.85 (d, J = 7.5Hz, 1H), 7.65 (d,J=5.9Hz,1H),7.58(d,J=7.6Hz,1H),7.52–7.44(m,1H),7.36–7.29(m,1H) ,6.86(d,J=9.2Hz,1H),4.38(s,1H),3.85(s,3H),3.82(s,2H),3.79–3.71(m, 1H),3.60–3.45(d,J=18.5Hz,3H),2.34–2.22(m,1H),2.12–2.01(m,1H).HRMS calcd.For C20H22N7O3S(M+H)+440.1499,found 440.1500.

[0229] Embodiment 18

[0230] (R)-methyl 4-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoate

[0231]

[0232] (1) Preparation of the title compound

[0233] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(4-(methoxycarbonyl)phenyl)acetic acid (37 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 41 mg of white solid powder with a yield of 58.3% and a melting point of 232-233°C.

[0234] 1 H NMR (400MHz, DMSO-d6) δ12.26(s,1H),8.50–8.45(m,1H),7.92(d,J=7.8Hz,2H),7.65(d,J=6.0Hz,1H),7.45(d,J=7.7Hz,2H),7.36–7.29(m,1 H),6.86(d,J=9.1Hz,1H),4.38(s,1H),3.84(s,3H),3.83(s,2H),3.79 –3.70(m,1H),3.60–3.46(m,3H),2.34–2.22(m,1H),2.13–2.00(m,1H).

[0235] HRMS calcd.For C20H22N7O3S(M+H)+440.1499,found 440.1497.

[0236] Embodiment 19

[0237] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid

[0238]

[0239] (1) Preparation of the title compound

[0240] Methyl (R)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoate (100 mg, 0.23 mmol) was dissolved in 1 mL of methanol, and 1 mL of sodium hydroxide solution (2 M) was added dropwise under ice bath. The mixture was moved to room temperature for 1 h. After the reaction was completed, 1 M hydrochloric acid was added under ice bath to quench the reaction to pH 4. The mixture was filtered to obtain 93 mg of white solid powder with a yield of 95.6% and a melting point of 182-183 ° C.

[0241] 1 H NMR (400MHz, Methanol-d4) δ8.57(s,1H),8.00(s,1H),7.97–7.88(m,2H),7.75(d,J=9.8Hz,1H),7.56(s,1H) ,7.50–7.43(m,1H),4.59(s,1H),4.02(s,1H),3.90(s,3H),3.33(d,J=14.5Hz,2H),2.55(s,1H),2.40(s,1H).

[0242] HRMS calcd.For C19H20N7O3S(M+H)+426.1343,found 426.1339.

[0243] Embodiment 20

[0244] (R)-4-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid

[0245]

[0246] (1) Preparation of the title compound

[0247] (R)-methyl 4-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoate (100 mg, 0.23 mmol) was dissolved in 1 mL of methanol, and 1 mL of sodium hydroxide solution (2 M) was added dropwise under ice bath. The mixture was moved to room temperature for 1 h. After the reaction was completed, 1 M hydrochloric acid was added under ice bath to quench the reaction to pH 4. The mixture was filtered to obtain 93 mg of white solid powder with a yield of 95.8% and a melting point of 182-183 ° C.

[0248] 1H NMR (400MHz, DMSO-d6) δ8.61(s,1H),7.89(d,J=7.9Hz,3H),7.78(d,J=9.6Hz,1H),7.42(d,J =7.9Hz,2H),4.57(s,1H),3.88(s,2H),3.77(s,3H),3.16(s,1H),2.34(s,1H),2.22(s,1H).

[0249] HRMS calcd.For C19H20N7O3S(M+H)+426.1343,found 426.1339.

[0250] Embodiment 21

[0251] (R)-3-(2-oxo-2-((5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoic acid ethyl ester

[0252]

[0253] (1) Preparation of the title compound

[0254] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), iodoethane (9 mg, 0.11 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated aqueous ammonium chloride solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, the organic phase was concentrated, and separated by column chromatography with the eluent being dichloromethane:methanol=10:1 to obtain 17 mg of white solid powder with a yield of 40.1% and a melting point of 175-176°C.

[0255] 1H NMR (400MHz, DMSO-d6) δ12.25(s,1H),8.47(d,J=4.4Hz,1H),7.92(s,1H),7.86(d,J=7.7Hz,1H) ,7.65(d,J=6.1Hz,1H),7.58(d,J=7.5Hz,1H),7.48(t,J=7.7Hz,1H),7.33(dd,J=9.2,4.4Hz,1H) ,6.87(d,J=9.2Hz,1H),4.40–4.31(m,1H),4.32(q,J=7.1Hz,2H),3.82(s,2H),3.74(dd,J=11.2, 5.8Hz,1H),3.59–3.47(m,3H),2.35–2.25(m,1H),2.10–1.99(m,1H),1.32(t,J=7.1Hz,3H).HRMS calcd.For C21H24N7O3S(M+H)+454.1656,found 454.1649.

[0256] Embodiment 22

[0257] (R)-isopropyl 3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoate

[0258]

[0259] (1) Preparation of the title compound

[0260] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), 2-iodopropane (12 mg, 0.11 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated aqueous ammonium chloride solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, the organic phase was concentrated, and separated by column chromatography with the eluent being dichloromethane:methanol=10:1 to obtain 17 mg of white solid powder with a yield of 39.5% and a melting point of 171-172°C.

[0261] 1H NMR(300MHz,DMSO-d6)δ12.28(s,1H),8.48(s,1H),7.95–7.80(m,2H),7.68(d ,J=5.9Hz,1H),7.60–7.55(m,1H),7.51–7.43(m,1H),7.39–7.29(m,1H),6.89 (d,J=8.7Hz,1H),5.21–5.06(m,1H),4.38(s,1H),3.81(s,2H),3.78–3.70(m, 1H), 3.62–3.45 (m, 3H), 2.37–2.20 (m, 1H), 2.09 (s, 1H), 1.31 (d, J = 6.1Hz, 6H).

[0262] HRMS calcd.For C22H26N7O3S(M+H)+468.1812,found 468.1810.

[0263] Embodiment 23

[0264] (R)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoic acid cyclopropylmethyl ester

[0265]

[0266] (1) Preparation of the title compound

[0267] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), chloromethylcyclopropane (10 mg, 0.11 mmol), sodium iodide (13 mg, 0.09 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated ammonium chloride aqueous solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, concentrated the organic phase, and separated by column chromatography. The eluent was dichloromethane:methanol.

[0268] =10:1, to obtain 19 mg of white solid powder, with a yield of 42.3% and a melting point of 166-167°C.

[0269] 1H NMR (400MHz, DMSO-d6) δ12.26(s,1H),8.47(s,1H),7.93(s,1H),7.87(d,J=7.6Hz,1H ),7.65(d,J=5.9Hz,1H),7.58(d,J=7.4Hz,1H),7.49(t,J=8.0Hz,1H),7.35–7.30(m, 1H),4.38(s,1H),4.16–4.10(m,2H),3.82(s,2H),3.75(s,1H),3.59–3.45(m,3H),2. 32–2.25(m,1H),2.12–2.04(m,1H),1.23(s,1H),0.56(d,J=6.6Hz,2H),0.35(s,2H).

[0270] HRMS calcd.For C23H26N7O3S(M+H)+480.1812,found 480.1808.

[0271] Embodiment 24

[0272] (R)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoic acid cyclopentyl ester

[0273]

[0274] (1) Preparation of the title compound

[0275] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), chloromethylcyclobutane (12 mg, 0.11 mmol), sodium iodide (13 mg, 0.09 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated ammonium chloride aqueous solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, concentrated the organic phase, and separated by column chromatography. The eluent was dichloromethane:methanol.

[0276] =10:1, to obtain 19 mg of white solid powder, with a yield of 41.6% and a melting point of 149-150°C.

[0277] 1H NMR (400MHz, DMSO-d6) δ12.28(s,1H),8.50(s,1H),7.94(s,1H),7.88(d,J=7.7Hz,1H),7.68(d, J=6.0Hz,1H),7.60(d,J=7.6Hz,1H),7.51(t,J=7.7Hz,1H),7.38–7.31(m,1H),6.89(d,J=9.1Hz, 1H),4.40(d,J=7.5Hz,1H),4.28(d,J=6.5Hz,2H),3.84(s,2H),3.77(dd,J=11.2,5.8Hz,1H),3.6 2–3.48(m,3H),2.79–2.67(m,1H),2.41–2.23(m,1H),2.17–2.01(m,3H),1.99–1.78(m,4H).HRMS calcd.For C24H28N7O3S(M+H)+494.1969,found 494.1966.

[0278] Embodiment 25

[0279] (R)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoic acid acetoxymethyl ester

[0280]

[0281] (1) Preparation of the title compound

[0282] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), chloromethyl acetate (12 mg, 0.11 mmol), sodium iodide (13 mg, 0.09 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated ammonium chloride aqueous solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, concentrated the organic phase, and separated by column chromatography. The eluent was dichloromethane:methanol.

[0283] =10:1, to obtain 20 mg of white solid powder, with a yield of 42.6% and a melting point of 170-172°C.

[0284] 1H NMR (700MHz, DMSO-d6) δ12.27(s,1H),8.48(d,J=4.4Hz,1H),7.95(s,1H),7.88(d,J=7.8Hz ,1H),7.66(d,J=6.0Hz,1H),7.63(d,J=7.7Hz,1H),7.52(t,J=7.7Hz,1H),7.36–7.31(m,1H ),6.88(d,J=9.3Hz,1H),5.93(s,2H),4.40–4.36(m,1H),3.83(s,2H),3.77–3.72(m,1H),3 .59–3.53(m,2H),3.50(d,J=8.1Hz,1H),2.32–2.25(m,1H),2.10(s,3H),2.08–2.05(m,1H).

[0285] HRMS calcd.For C22H24N7O5S(M+H)+498.1554,found 498.1556.

[0286] Embodiment 26

[0287] (R)-2-(Dimethylamino)-2-oxoethyl 3-(2-oxo-2-(5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoate

[0288]

[0289] (1) Preparation of the title compound

[0290] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), 2-chloro-N,N-dimethylacetamide (14 mg, 0.11 mmol), sodium iodide (13 mg, 0.09 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated aqueous ammonium chloride solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, concentrated the organic phase, separated by column chromatography, the eluent was dichloromethane:methanol=10:1, and 19 mg of yellow solid powder was obtained, the yield was 39.7%, and the melting point was 140-142°C.

[0291] 1H NMR (400MHz, DMSO-d6) δ12.29(s,1H),8.49(d,J=4.4Hz,1H),7.97(s,1H),7.90(d,J=7.7Hz, 1H),7.68(d,J=6.0Hz,1H),7.62(d,J=7.6Hz,1H),7.53(t,J=7.7Hz,1H),7.35(dd,J=9.1,4. 4Hz,1H),6.89(d,J=9.1Hz,1H),5.05(s,2H),4.40(s,1H),3.85(s,2H),3.77(dd,J=11.1,5. 8Hz,1H),3.64–3.48(m,3H),3.00(s,3H),2.86(s,3H),2.35–2.25(m,1H),2.14–2.04(m,1H).

[0292] HRMS calcd.For C23H27N8O4S(M+H)+511.1870,found 511.1871.

[0293] Embodiment 27

[0294] (R)-(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl 3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoate

[0295]

[0296] (1) Preparation of the title compound

[0297] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), 4-chloromethyl-5-methyl-1,3-dioxol-2-one (17 mg, 0.11 mmol), sodium iodide (13 mg, 0.09 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated aqueous ammonium chloride solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, concentrated the organic phase, separated by column chromatography, the eluent was dichloromethane:methanol=10:1, and 25 mg of yellow solid powder was obtained, the yield was 49.3%, and the melting point was 145-146°C.

[0298] 1H NMR (400MHz, DMSO-d6) δ12.28(s,1H),8.50(d,J=4.4Hz,1H),7.97(s,1H),7.90(d,J=7. 7Hz,1H),7.67(d,J=6.0Hz,1H),7.63(d,J=7.7Hz,1H),7.52(t,J=7.7Hz,1H),7.40–7.33 (m,1H),6.91(d,J=9.1Hz,1H),5.25(s,2H),4.40(s,1H),3.85(s,2H),3.77(dd,J=11.3, 5.8Hz,1H),3.60–3.50(m,3H),2.38–2.30–2.20(m,1H),2.24(s,3H),2.13–2.03(m,1H).

[0299] HRMS calcd.For C24H24N7O6S(M+H)+538.1503,found 538.1504.

[0300] Embodiment 28

[0301] (R)-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzoic acid (ethoxycarbonyl)oxy)methyl ester

[0302]

[0303] (1) Preparation of the title compound

[0304] (R)-3-(2-oxo-2-(5-(1-(pyridazin-3-yl)pyrrolidin-3-ylamino)-1,3,4-thiadiazol-2-yl)amino)ethylbenzoic acid (40 mg, 0.09 mmol), chloromethyl ethyl carbonate (15 mg, 0.11 mmol), sodium iodide (13 mg, 0.09 mmol) and cesium carbonate (122 mg, 0.376 mmol) were dissolved in 1 mL of N,N-dimethylformamide and reacted at 60°C for 5 h. The reaction was quenched by adding saturated ammonium chloride aqueous solution, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol.

[0305] =10:1, to obtain 23 mg of yellow solid powder, with a yield of 46.3% and a melting point of 160-162°C.

[0306] 1H NMR (400MHz, DMSO-d6) δ12.26(s,1H),8.47(d,J=4.4Hz,1H),7.96(s,1H),7.89(d,J=7.8Hz,1H ),7.65(t,J=7.5Hz,2H),7.52(t,J=7.7Hz,1H),7.33(dd,J=9.2,4.4Hz,1H),6.88(d,J=9.1Hz,1 H),5.95(s,2H),4.38(d,J=7.3Hz,1H),4.19(q,J=7.1Hz,2H),3.84(s,2H),3.75(dd,J=11.1,5 .8Hz,1H),3.60–3.46(m,3H),2.35–2.22(m,1H),2.13–2.02(m,1H),1.22(t,J=7.1Hz,3H).HRMS calcd.For C23H26N7O6S(M+H)+528.1660,found 528.1664.

[0307] Embodiment 29

[0308] (R)-2-(3-(1H-tetrazol-5-yl)phenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazole-

[0309] 2-amino)acetamide

[0310]

[0311] (1) Preparation of methyl 2-(3-cyanophenyl)acetate

[0312]

[0313] Add methyl 2-(3-bromophenyl)acetate (1.00 g, 4.37 mmol), zinc cyanide (510 mg, 4.37 mmol), tris(dibenzylideneacetone)dipalladium (305 mg, 0.33 mmol), 1,1'-bis(diphenylphosphino)ferrocene (365 mg, 0.65 mmol) and zinc powder (15 mg, 0.22 mmol) into a bottle in sequence, dissolve in 10 mL of N,N-dimethylacetamide, replace argon, and heat to 110 ° C for 3 h. Cool the reaction to room temperature, add water to quench the reaction, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry it over anhydrous sodium sulfate. Concentrate and separate by column chromatography, the eluent is petroleum ether: ethyl acetate = 5:1, and obtain 563 mg of light yellow oil with a yield of 73.6%.

[0314] 1H NMR (400MHz, Chloroform-d) δ7.61–7.55(m,2H),7.53(d,J=7.5Hz,1H),7.48–7.40(m,1H),3.73(s,3H),3.68(s,2H).

[0315] MS (ESI) m / z 176.11 (M+H) + .

[0316] (2) Preparation of methyl 2-(3-(1H-tetrazol-5-yl)phenyl)acetate

[0317]

[0318] Add methyl 2-(3-cyanophenyl)acetate (1.00 g, 5.71 mmol), sodium azide (557 mg, 8.56 mmol) and cuprous iodide (272 mg, 1.43 mmol) to a bottle in sequence, dissolve in 10 mL of N,N-dimethylformamide, replace argon, and heat to 120 ° C for 3 h. Cool the reaction to room temperature, add water to quench the reaction, add 1 M hydrochloric acid to adjust pH 2, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Concentrate, separate by column chromatography, eluent is petroleum ether: ethyl acetate = 1:1, and obtain 1.08 g of white solid, yield 86.6%, melting point 143-144 ° C.

[0319] 1 H NMR (400MHz, Chloroform-d) δ7.98(s,1H),7.88(s,1H),7.41(s,2H),3.81–3.75(m,5H).

[0320] MS (ESI) m / z 219.07 (M+H) + .

[0321] (3) Preparation of 2-(3-(1H-tetrazol-5-yl)phenyl)acetic acid

[0322]

[0323] Dissolve methyl 2-(3-(1H-tetrazolyl-5-yl)phenyl)acetate (1.00 g, 4.58 mmol) in 5 mL of methanol, add 5 mL of 2M sodium hydroxide solution dropwise under ice bath, react at room temperature for 2 h, remove the methanol solvent by vacuum distillation, add 1M hydrochloric acid to adjust pH 2, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Remove the solvent by vacuum distillation to obtain 921 mg of a white solid, with a yield of 98.6% and a melting point of 191-192°C.

[0324] 1 H NMR (400MHz, DMSO-d6) δ7.96 (s, 1H), 7.92 (d, J = 7.7Hz, 1H), 7.56 (t, J = 7.7Hz, 1H), 7.49 (d, J = 7.6Hz, 1H), 3.71 (s, 2H).

[0325] MS (ESI) m / z 205.10 (M+H) + .

[0326] (4) Preparation of the title compound

[0327] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-(1H-tetrazolyl-5-yl)phenyl)acetic acid (39 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphoric anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 43 mg of white solid powder with a yield of 60.5% and a melting point of 242-243°C.

[0328] 1 H NMR (400MHz, DMSO-d6) δ8.49(d,J=4.6Hz,1H),8.05(s,1H),7.97(d,J=7.4Hz,1H),7.73(s,1H),7.51(d,J=7.6Hz,1H),7.46 (s,1H),7.35(s,1H),6.89(d,J=9.1Hz,1H),4.40(s,1H),3.85(s,2H),3.76(s,1H),3.58(s,3H),2.30(s,1H),2.10(s,1H).

[0329] HRMS calcd.For C19H20N11OS(M+H)+450.1568,found 450.1576.

[0330] Embodiment 30

[0331] (R)-N-Methylsulfonyl-3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)benzamide

[0332]

[0333] (1) Preparation of 3-(2-methoxy-2-oxoethyl)benzoic acid

[0334]

[0335] Dissolve 3-carboxyphenylacetic acid (1.00 g, 5.55 mmol) in 10 mL of anhydrous methanol, add 2 drops of thionyl chloride, and react at room temperature for 1 h. The white solid is 1.07 g, with a yield of 99.9% and a melting point of 72-73°C.

[0336] 1 H NMR (400MHz, Chloroform-d) δ8.06 (s, 2H), 7.57 (d, J = 7.5Hz, 1H), 7.48 (d, J = 7.6Hz, 1H), 3.74 (d, J = 5.0Hz, 5H).

[0337] MS (ESI) m / z 195.07 (M+H) + .

[0338] (2) Preparation of methyl 2-(3-((methylsulfonyl)carbamoyl)phenyl)acetate

[0339]

[0340] 3-(2-methoxy-2-oxoethyl)benzoic acid (1.00 g, 5.15 mmol), methanesulfonamide (980 mg, 10.30 mmol), HATU condensing agent (2.94 g, 7.72 mmol) and 4-dimethylaminopyridine (322 mg, 2.58 mmol) were added to a bottle in sequence, dissolved in 10 mL of dichloromethane, stirred for 5 minutes, and triethylamine (1.04 g, 10.30 mmol) was added dropwise. After reacting at room temperature for 1 hour, 1M hydrochloric acid was used to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic layer was washed twice with a saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentrated, separated by column chromatography, the eluent was petroleum ether: ethyl acetate = 2:1, and 893 mg of light yellow oil was obtained, with a yield of 64.0%.

[0341] 1 H NMR (400MHz, Chloroform-d) δ9.02 (s, 1H), 7.76 (d, J = 10.7Hz, 2H), 7.53 (d, J = 7. 7Hz,1H),7.45(t,J=6.7Hz,1H),3.72(s,3H),3.71(d,J=2.4Hz,2H),3.44(s,3H).

[0342] MS (ESI) m / z 271.98 (M+H) + .

[0343] (2) Preparation of 2-(3-((methylsulfonyl)carbamoyl)phenyl)acetic acid

[0344]

[0345] Dissolve methyl 2-(3-((methylsulfonyl)carbamoyl)phenyl)acetate (1.00 g, 3.69 mmol) in 5 mL of methanol, add 5 mL of 2M sodium hydroxide solution dropwise under ice bath, react at room temperature for 2 h, remove the methanol solvent by vacuum distillation, add 1M hydrochloric acid to adjust pH to 2, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Remove the solvent by vacuum distillation to obtain 945 mg of a white solid with a yield of 99.6% and a melting point of 188-189°C.

[0346] 1 H NMR (400MHz, Methanol-d4) δ7.84–7.76(m,2H),7.60–7.54(m,1H),7.50–7.44(m,1H),3.71(s,2H),3.36(s,3H).

[0347] MS (ESI) m / z 257.94 (M+H) + .

[0348] (3) Preparation of the title compound

[0349] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-((methylsulfonyl)carbamoyl)phenyl)acetic acid (49 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, the reaction mixture was moved to room temperature for 4 h, after which the reaction was quenched with water, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, the organic phase was concentrated, and separated by column chromatography with the eluent being dichloromethane:methanol = 10:1 to give 32 mg of a white solid powder with a yield of 39.4% and a melting point of 206-208°C.

[0350] 1H NMR (400MHz, DMSO-d6) δ12.28(s,1H),8.50(s,1H),7.90(s,1H),7.85(d,J=7.0Hz,1H),7.67(s,1H),7.57(d,J=7.5Hz,1H),7.49(d,J=8.0Hz,1H),7. 37(s,1H),6.93(d,J=9.3Hz,1H),4.40(s,1H),3.82(s,2H),3.79–3.72(m, 1H),3.63–3.48(m,3H),3.34(s,3H),2.35–2.24(m,1H),2.15–2.04(m,1H).

[0351] HRMS calcd.For C20H23N8O4S2(M+H)+503.1278,found 503.1283.

[0352] Embodiment 31

[0353] (R)-Diethyl(3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)phenyl)phosphonate

[0354]

[0355] (1) Preparation of 2-(3-bromophenyl)benzyl acetate

[0356]

[0357] 3-Bromophenylacetic acid (1 g, 4.67 mmol) was dissolved in 10 mL of anhydrous dichloromethane, benzyl alcohol (2.99 g, 27.75 mmol) was added, and 2 drops of thionyl chloride were added dropwise. The mixture was reacted at room temperature for 3 h in a dry environment until the insoluble solid disappeared from the solution. The solvent was removed by distillation under reduced pressure, and a saturated aqueous ammonium chloride solution was added to quench the reaction. The mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was petroleum ether:ethyl acetate = 4:1 to obtain 1.38 g of colorless oil with a yield of 97.3%.

[0358] 1 H NMR (400MHz, Chloroform-d) δ7.44(s,1H),7.44–7.36(m,1H),7.37–7.29(m,5H),7.24–7.15(m,2H),5.14(s,2H),3.63(s,2H).

[0359] MS (ESI) m / z 305.01 (M+H) + .

[0360] (2) Preparation of 2-(3-(diethoxyphospho)phenyl)benzyl acetate

[0361]

[0362] 2-(3-bromophenyl) benzyl acetate (1.00 g, 3.29 mmol), triethyl phosphite (600 mg, 3.62 mmol) and palladium acetate (161 mg, 0.91 mmol) were added to a microwave reaction tube in sequence, dissolved in 8 mL of anhydrous acetonitrile, protected by argon, microwaved at 160°C for 30 min, quenched with water, extracted with ethyl acetate three times, washed the organic layer twice with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated, separated by column chromatography, the eluent was petroleum ether: ethyl acetate = 2:1, and 810 mg of light yellow oil was obtained, with a yield of 67.9%.

[0363] 1 H NMR(400MHz,Chloroform-d)δ7.73(d,J=12.9Hz,2H),7.52–7.40(m,2H),7.32(d, J=6.4Hz,5H),5.14(s,2H),4.17–4.03(m,4H),3.72(s,2H),1.31(t,J=7.1Hz,6H).

[0364] MS (ESI) m / z 363.10 (M+H) + .

[0365] (3) Preparation of 2-(3-(diethoxyphospho)phenyl)acetic acid

[0366]

[0367] Benzyl 2-(3-(diethoxyphosphoryl)phenyl)acetate (1.00 g, 2.76 mmol) was dissolved in 10 mL of methanol, Raney Ni was added, hydrogen was introduced, and the reaction was carried out at room temperature for 2 h. After the catalyst was filtered off, the solvent was removed by distillation under reduced pressure to obtain 750 mg of a light yellow oil with a yield of 99.9%.

[0368] 1 H NMR (400MHz, Chloroform-d) δ7.82 (d, J = 13.9 Hz, 1H), 7.67 (s, 1H), 7.46 (m, 2H), 4.13 (s, 4H), 3.69 (s, 2H), 1.31 (s, 6H).

[0369] MS (ESI) m / z 272.89 (M+H) + .

[0370] (4) Preparation of the title compound

[0371] (R)-N 2 -(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-(diethoxyphosphoryl)phenyl)acetic acid (52 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphoric anhydride in ethyl acetate was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 37 mg of white solid powder with a yield of 44.6% and a melting point of 152-153°C.

[0372] 1 H NMR (400MHz, DMSO-d6) δ12.26 (s, 1H), 8.47 (d, J = 4.1Hz, 1H), 7.72–7.64 (m, 2H), 7.61 (t,J=6.5Hz,1H),7.58–7.53(m,1H),7.51(dd,J=7.6,4.4Hz,1H),7.37–7.29(m,1H),6 .87(d,J=9.3Hz,1H),4.38(s,1H),4.01(m,4H),3.81(s,2H),3.75(dd,J=11.1,5.8Hz ,1H),3.61–3.46(m,3H),2.35–2.22(m,1H),2.11–2.02(m,1H),1.22(t,J=7.0Hz,6H).

[0373] HRMS calcd.For C22H29N7O4PS(M+H)+518.1734,found 518.1732.

[0374] Embodiment 32

[0375] (R)-(3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)phenyl)phosphonic acid

[0376]

[0377] (1) Preparation of the title compound

[0378] (R)-(3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)phenyl)phosphonic acid diethyl ester (100 mg, 0.19 mmol) was dissolved in 1 mL of a mixed solvent of dichloromethane:N,N-dimethylformamide = 1:10, and trimethylsilyl bromide (297 mg, 1.93 mmol) was added dropwise. The mixture was reacted at room temperature for 4 h. After the reaction was completed, ethyl acetate was added to quench the reaction. The precipitated solid was filtered off with suction, and the filter cake was washed with ethyl acetate:methanol = 20:1 to obtain 75 mg of white solid powder with a yield of 85.3% and a melting point of 267-268°C.

[0379] 1 H NMR (400MHz, DMSO-d6) δ8.60(s,1H),7.92(d,J=6.2Hz,1H),7.80(d,J=9.6Hz,1H),7.65(d,J=13.5Hz,1H),7.60–7.53 (m,1H),7.45–7.39(m,2H),4.47(s,1H),3.92–3.84(m,1H),3.79(s,2H),3.72(s,3H),2.40–2.30(m,1H),2.19(s,1H).

[0380] HRMS calcd.For C18H21N7O4PS(M+H)+462.1108,found 462.1105.

[0381] Embodiment 33

[0382] (R)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetamide

[0383]

[0384] (1) Preparation of 2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)benzyl acetate

[0385]

[0386] 2-(3-bromophenyl) benzyl acetate (1.00 g, 3.29 mmol), biboronic acid pinacol ester (920 mg, 3.62 mmol), DPPF palladium dichloride (480 mg, 0.66 mmol) and potassium acetate (580 mg, 5.92 mmol) were added to the reaction bottle in sequence, dissolved in 10 mL of anhydrous 1,4-dioxane, argon was replaced, heated at 80°C for 6 h, quenched with saturated ammonium chloride aqueous solution, extracted with ethyl acetate three times, the organic layer was washed twice with saturated sodium chloride solution, and dried over anhydrous sodium sulfate. Concentrated, separated by column chromatography, the eluent was petroleum ether: ethyl acetate = 4:1, and 891 mg of light yellow oil was obtained, with a yield of 76.9%.

[0387] 1 H NMR (400MHz, DMSO-d6) δ7.67–7.50(m,2H),7.41–7.30(m,7H),5.10(s,2H),3.76(s,2H),1.29(s,12H).

[0388] MS (ESI) m / z 353.05 (M+H) + .

[0389] (2) Preparation of 2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetic acid

[0390]

[0391] Dissolve 2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)benzyl acetate (1.00 g, 2.84 mmol) in 10 mL of methanol, add Raney Ni, introduce hydrogen, and react at room temperature for 2 h. After filtering the catalyst, remove the solvent by distillation under reduced pressure to obtain 724 mg of light yellow oil with a yield of 97.3%.

[0392] 1 H NMR (400MHz, DMSO-d6) δ7.59–7.50(m,2H),7.40–7.27(m,2H),3.57(s,2H),1.29(s,12H).

[0393] MS (ESI) m / z 263.04 (M+H) + .

[0394] (3) Preparation of the title compound

[0395] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetic acid (50 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 46 mg of white solid powder with a yield of 56.2% and a melting point of 255-256°C.

[0396] 1 H NMR (400MHz, DMSO-d6) δ12.24(s,1H),8.50(d,J=4.4Hz,1H),7.67(d,J=6.0Hz,2H),7.58(d,J=7.4Hz,1H),7.45(d,J=7.7Hz,1H),7.40 –7.31(m,2H),6.89(d,J=9.1Hz,1H),4.40(s,1H),3.74(s,2H),3.62–3.48(m,3H),2.39–2.24(m,1H),2.13–2.06(m,1H),1.32(s,12H).

[0397] HRMS calcd.For C24H31BN7O3S(M+H)+508.2297,found 508.2304.

[0398] Embodiment 34

[0399] (R)-(3-(2-oxo-2-((5-((1-(pyridazin-3-yl)pyrrolidin-3-yl)amino)-1,3,4-thiadiazol-2-yl)amino)ethyl)phenyl)boronic acid

[0400]

[0401] (1) Preparation of the title compound

[0402] (R)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetamide (100 mg, 0.20 mmol), sodium periodate (64 mg, 0.30 mmol) and ammonium acetate (24 mg, 0.30 mmol) were dissolved in 1 mL of a mixed solvent of acetone: water: N,N-dimethylformamide = 1:1:8, and reacted at 50°C for 48 h. After the reaction was completed, water was added to quench the reaction, the precipitated solid was filtered, and the filter cake was washed with ethyl acetate: methanol = 20:1 to obtain 38 mg of white solid powder with a yield of 45.9% and a melting point of 265-266°C.

[0403] 1 H NMR (400MHz, Methanol-d4) δ8.43 (s, 1H), 7.56 (d, J = 22.8Hz, 2H), 7.37 (s, 3H), 6.98 (d, J = 9.5Hz, 1H),4.49(s,1H),3.85(s,1H),3.77(s,2H),3.66(s,2H),3.60(s,1H),2.40(s,1H),2.21(s,1H).

[0404] HRMS calcd.For C18H21BN7O3S(M+H)+426.1514,found 426.1513.

[0405] Embodiment 35

[0406] (R)-2-(3-phenoxyphenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0407]

[0408] (1) Preparation of the title compound

[0409] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(3-phenoxyphenyl)acetic acid (43 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, and N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise. The reaction solution was cooled to 0°C, and 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise. The mixture was moved to room temperature and reacted for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 48 mg of white solid powder with a yield of 63.8% and a melting point of 215-216°C.

[0410] 1 H NMR (400MHz, DMSO-d6) δ12.20(s,1H),8.50(d,J=4.4Hz,1H),7.67(d,J=6.1Hz,1H),7.4 2(t,J=7.7Hz,2H),7.35(dd,J=9.2,5.3Hz,2H),7.17(t,J=7.4Hz,1H),7.09(d,J=7.6Hz, 1H),7.04(d,J=8.0Hz,1H),7.00(s,2H),6.90(t,J=8.4Hz,1H),4.40(s,1H),3.77(dd,J= 11.3,5.9Hz,1H),3.73(s,2H),3.61–3.48(m,3H),2.38–2.25(m,1H),2.15–2.05(m,1H).

[0411] HRMS calcd.For C24H24N7O2S(M+H)+474.1707,found 474.1707.

[0412] Embodiment 36

[0413] (R)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)-2-(3-o-tolylphenyl)acetamide

[0414]

[0415] (1) Preparation of the title compound

[0416] (R)-2-(3-bromophenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide (100 mg, 0.22 mmol), 2-methylphenol (36 mg, 0.33 mmol), cuprous iodide (22 mg, 0.11 mmol), cesium carbonate (214 mg, 0.65 mmol) and N,N-dimethylglycine (60 mg, 0.44 mmol) were added to a microwave reaction tube in sequence, dissolved in 1 mL of 1,4-dioxane, protected by argon, and subjected to microwave reaction at 90°C for 3 h. After the reaction was completed, water was added to quench the reaction, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, concentrated the organic phase, and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 14 mg of white solid powder with a yield of 12.7% and a melting point of 201-202°C.

[0417] 1 H NMR (400MHz, DMSO-d6) δ12.20(s,1H),7.67(s,1H),7.37–7.28(m,3H),7.23(d,J=7.7Hz,1H),7.13(t,J=7.4Hz,2H),7.02(d,J=7.5Hz,1H),6.96– 6.88(m,3H),6.77(d,J=8.2Hz,1H),4.40(s,1H),3.78(s,1H),3.70(s,2H ),3.63–3.49(m,3H),2.37–2.28(m,1H),2.18(s,3H),2.14–2.06(m,1H).

[0418] HRMS calcd.For C25H26N7O2S(M+H)+488.1863,found 488.1862.

[0419] Embodiment 37

[0420] (R)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)-2-(3-o-tolylphenyl)acetamide

[0421]

[0422] (1) Preparation of the title compound

[0423] (R)-2-(3-bromophenyl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide (100 mg, 0.22 mmol), 2-benzyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (118 mg, 0.55 mmol), DPPF palladium dichloride (60 mg, 0.11 mmol) and potassium carbonate (60 mg, 0 .44mmol) were added to a microwave reaction tube in sequence, dissolved in 1mL of a mixed solvent of 1,4-dioxane: water = 3:1, protected by argon, and microwaved at 80°C for 6h. After the reaction, water was added to quench the reaction, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, and the organic phase was concentrated and separated by column chromatography with an eluent of dichloromethane: methanol = 10:1 to obtain 22mg of a white solid powder with a yield of 21.6% and a melting point of 210-211°C.

[0424] 1 H NMR (400MHz, DMSO-d6) δ12.19(s,1H),8.50(s,1H),7.66(d,J=6.0Hz,1H),7.38–7.22(m,6H),7.19(s,2H),7.13(d,J=8.3Hz,2H),6.89( d,J=9.0Hz,1H),4.40(s,1H),3.94(s,2H),3.76(d,J=6.2Hz,1H),3.69(s,2H),3.55(d,J=18.9Hz,3H),2.31(s,1H),2.17–1.95(m,1H).

[0425] HRMS calcd.For C25H26N7OS(M+H)+472.1914,found 472.1914.

[0426] Embodiment 38

[0427] (R)-2-([1,1'-biphenyl]-3-yl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0428]

[0429] (1) Preparation of the title compound

[0430] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-phenylphenylacetic acid (43 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 41 mg of white solid powder with a yield of 55.4% and a melting point of 183-184°C.

[0431] 1 H NMR (400MHz, DMSO-d6) δ12.25(s,1H),8.49(dd,J=4.5,1.2Hz,1H),7.66(d,J=6.9Hz,3H),7 .63(s,1H),7.61–7.54(m,1H),7.49(dd,J=8.4,6.8Hz,2H),7.44(t,J=7.7Hz,1H),7.39(t, J=7.3Hz,1H),7.38–7.29(m,2H),6.88(dd,J=9.1,1.3Hz,1H),4.43–4.36(m,1H),3.82(s,2 H),3.77(dd,J=11.0,5.8Hz,1H),3.62–3.48(m,3H),2.37–2.24(m,1H),2.15–2.03(m,1H).

[0432] HRMS calcd.For C24H24N7OS(M+H)+458.1758,found 458.1756.

[0433] Embodiment 39

[0434] (R)-2-(Furan-2-yl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0435]

[0436] (1) Preparation of the title compound

[0437] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), furan-2-acetic acid (24 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 35 mg of white solid powder with a yield of 58.6% and a melting point of 240-242°C.

[0438] 1 H NMR (400MHz, DMSO-d6) δ12.22(s,1H),8.50(d,J=4.4Hz,1H),7.69(d,J=6.1Hz,1H),7.59(s,1H),7.37(dd,J=9.2,4.4Hz,1H),6.92(d,J=9.2Hz,1H), 6.42(s,1H),6.29(d,J=3.2Hz,1H),4.41(s,1H),3.84(s,2H),3.78(dd,J= 11.1,5.8Hz,1H),3.65–3.47(m,3H),2.38–2.25(m,1H),2.17–2.04(m,1H).

[0439] HRMS calcd.For C16H18N7O2S(M+H)+372.1237,found 372.1235.

[0440] Embodiment 40

[0441] (R)-2-(Thiophen-2-yl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0442]

[0443] (1) Preparation of the title compound

[0444] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), thiophene-2-acetic acid (27 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 30 mg of white solid powder with a yield of 48.6% and a melting point of 232-234°C.

[0445] 1 H NMR (400MHz, DMSO-d6) δ12.24(s,1H),8.50(d,J=4.4Hz,1H),7.69(d,J=6.0Hz,1H),7.44–7.41(m,1H),7.36(dd,J=9.2,4.4Hz,1H),6.99(d,J=3 .4Hz,2H),6.90(d,J=9.1Hz,1H),4.40(d,J=7.3Hz,1H),3.77(dd,J=11.0,5.7Hz,1H),3.65–3.49(m,3H),2.36–2.25(m,1H),2.15–2.05(m,1H).

[0446] HRMS calcd.For C16H18N7OS2(M+H)+388.1009,found 388.1006.

[0447] Embodiment 41

[0448] (R)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)-2-(pyridin-2-yl)acetamide

[0449]

[0450] (1) Preparation of the title compound

[0451] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), pyridine-2-acetic acid (26 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 32 mg of white solid powder with a yield of 52.8% and a melting point of 140-141°C.

[0452] 1 H NMR (400MHz, DMSO-d6) δ12.24(s,1H),8.50(s,2H),7.78(t,J=7.6Hz,1H),7.67(d,J=6.1Hz,1H),7.41–7.33(m,2H),7.33–7.26(m,1H),6.91(d HRMS calcd.For C17H19N8OS(M+H)+383.1397,found383.1389.

[0453] Embodiment 42

[0454] (R) 2-(Naphthalen-2-yl)-N-(5-(1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)acetamide

[0455]

[0456] (1) Preparation of the title compound

[0457] (R)-N 2-(1-(pyridazine-3-yl)pyrrolidin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 2-(naphthalene-2-yl)acetic acid (35 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 40 mg of white solid powder with a yield of 57.6% and a melting point of 267-268°C. 1 H NMR(400MHz,DMSO-d6)δ12.30(s,1H),8.49(s,1H),7.93–7.88(d,J=8.0Hz ,3H),7.83(s,1H),7.66(d,J=6.0Hz,1H),7.56–7.45(m,3H),7.34(dd,J=8. 9,4.4Hz,1H),6.88(d,J=9.1Hz,1H),4.40(s,1H),3.92(s,2H),3.76(dd,J= 11.1,5.8Hz,1H),3.62–3.49(m,3H),2.37–2.24(m,1H),2.13–2.04(s,1H).

[0458] HRMS calcd.For C22H22N7OS(M+H)+432.1601,found 432.1597.

[0459] Embodiment 43

[0460] (S)-2-methoxy-2-phenyl-N-(5-((R)-1-(pyridazin-3-ylamino)propyl-2-amino)-1,3,4-thiadiazol-2-yl)

[0461] Acetamide

[0462]

[0463] (1) Preparation of tert-butyl (R)-(1-((6-chloropyridazin-3-yl)amino)propan-2-yl)carbamate

[0464]

[0465] 3,6-dichloropyridazine (1.00 g, 6.71 mmol), (R)-(1-aminopropyl-2-yl)carbamic acid tert-butyl ester (1.40 g, 8.06 mmol) and N,N-diisopropylethylamine (1.04 g, 8.06 mmol) were added to the bottle in sequence, dissolved in 10 mL of acetonitrile, and heated to 80 ° C for 5 h. The reaction was cooled to room temperature, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic layer was washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentration and column chromatography were performed, and the eluent was petroleum ether: ethyl acetate = 1:1 to obtain 1.62 g of white solid, with a yield of 84.3% and a melting point of 155-156 ° C.

[0466] 1 H NMR (400MHz, Chloroform-d) δ7.27 (d, J = 2.2Hz, 1H), 7.14 (d, J = 10.4Hz, 1H), 6.72 (d, J = 9. 2Hz,1H),4.68(s,1H),3.96–3.88(m,1H),3.48(s,2H),1.42(s,9H),1.24(d,J=6.4Hz,3H).

[0467] MS (ESI) m / z 287.03 (M+H) + .

[0468] (2) Preparation of tert-butyl (R)-(1-((pyridazin-3-yl)amino)propan-2-yl)carbamate

[0469]

[0470] Dissolve (R)-tert-butyl (1-((6-chloropyridazin-3-yl)amino)propan-2-yl)carbamate (1.00 g, 3.49 mmol), ammonium formate (2.19 g, 34.90 mmol) and palladium / carbon (100 mg, 10%) in 10 mL of ethanol and react at room temperature for 3 h. Filter the insoluble solids through diatomaceous earth and distill the filtrate under reduced pressure to obtain 877 mg of a white solid with a yield of 99.6% and a melting point of 169-170°C.

[0471] 1 H NMR (400MHz, Chloroform-d) δ8.52(s,1H),7.15(s,1H),6.72(s,1H),5.72(s,1H),4.83(s,1H),3.94(s,1H),3.50(s,2H),1.42(s,9H),1.25(s,3H).

[0472] MS (ESI) m / z 253.10 (M+H) + .

[0473] (3)(R)-N 2 Preparation of -(1-(pyridazin-3-ylamino)propyl-2-yl)-1,3,4-thiadiazole-2,5-diamine

[0474]

[0475] Dissolve (R)-tert-butyl (1-((pyridazin-3-yl)amino)propan-2-yl)carbamate (1.00 g, 3.96 mmol) in 10 mL of dichloromethane, add 5 mL of trifluoroacetic acid dropwise in an ice-water bath, transfer to room temperature for reaction for 1 h, remove the solvent by distillation under reduced pressure, dissolve in isopropanol solution, add 5-bromo-1,3,4-thiadiazole-2-amine (855 mg, 4.76 mmol) and N,N-diisopropylethylamine (615 mg, 4.76 mmol), mix well, transfer to 45 ° C and heat for reaction for 4 h, remove the solvent by distillation under reduced pressure, add water to quench the reaction, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Concentrate and separate by column chromatography, the eluent is dichloromethane:methanol = 10:1. Obtain 729 mg of off-white solid, yield 73.3%, melting point 210-211 ° C.

[0476] 1 H NMR (400MHz, Methanol-d4) δ8.37(s,1H),7.29(s,1H),6.97(d,J=9.1Hz,1H),3.93(h,J=6.7Hz,1H),3.59–3.47(m,2H),1.29(s,3H).

[0477] MS (ESI) m / z 251.98 (M+H) + .

[0478] (4) Preparation of the title compound

[0479] (R)-N 2 -(1-(pyridazin-3-ylamino)propyl-2-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), (S)-2-methoxy-2-phenylacetic acid (32 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 37 mg of white solid powder with a yield of 57.6% and a melting point of 181-182°C.

[0480] 1 H NMR (400MHz, DMSO-d6) δ12.13(s,1H),8.40(s,1H),7.45(s,2H),7.37(d,J=9.3Hz,4H),7.19(s,1H),6.90 (s,1H),6.83(d,J=9.1Hz,1H),4.97(s,1H),3.95(s,1H),3.46(s,3H),3.30(s,2H),1.19(d,J=6.4Hz,3H).

[0481] HRMS calcd.For C18H22N7O2S(M+H)+400.1477,found 400.1475.

[0482] Embodiment 44

[0483] (R)-N-(5-(1-(pyridazin-3-ylamino)propyl-2-amino)-1,3,4-thiadiazol-2-yl)-2-(3-(trifluoromethoxy)phenyl)acetamide

[0484]

[0485] (1) Preparation of the title compound

[0486] (R)-N 2 -(1-(pyridazin-3-ylamino)propyl-2-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.16 mmol), 3-trifluoromethoxyphenylacetic acid (42 mg, 0.19 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 140 μL of 50% propylphosphonic anhydride in ethyl acetate solution was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 45 mg of white solid powder with a yield of 62.3% and a melting point of 225-226°C.

[0487] 1 H NMR(400MHz,Methanol-d4)δ8.35(s,1H),7.43(s,1H),7.33(s,1H),7.26(s,2H),7.20( s,1H),6.93(d,J=9.2Hz,1H),4.04(s,1H),3.80(s,2H),3.64–3.47(m,2H),1.29(s,3H).

[0488] HRMS calcd.For C18H19F3N7O2S(M+H)+454.1195,found 454.1196.

[0489] Embodiment 45

[0490] (S)-2-methoxy-2-phenyl-N-(5-(((R)-1-((6-(trifluoromethyl)pyridazin-3-yl)amino)propan-2-yl)amino)-

[0491] 1,3,4-thiadiazol-2-yl)acetamide

[0492]

[0493] (1) Preparation of tert-butyl (R)-(1-((6-(trifluoromethyl)pyridazin-3-yl)amino)propan-2-yl)carbamate

[0494]

[0495] 3-Chloro-6-trifluoromethylpyridazine (1.00 g, 5.49 mmol), (R)-(1-aminopropyl-2-yl)carbamic acid tert-butyl ester (927 mg, 5.39 mmol) and N,N-diisopropylethylamine (696 mg, 5.39 mmol) were added to the bottle in sequence, dissolved in 10 mL of acetonitrile, and heated to 80 ° C for 5 h. The reaction was cooled to room temperature, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic layer was washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentration and column chromatography were performed, and the eluent was petroleum ether: ethyl acetate = 1:1 to obtain 1.43 g of white solid, with a yield of 81.6% and a melting point of 144-145 ° C.

[0496] 1 H NMR(400MHz,Chloroform-d)δ7.43(d,J=9.4Hz,1H),6.84(d,J=9.3Hz,1H),4.77 (s,1H),3.95(t,J=6.9Hz,1H),3.56(s,2H),1.41(s,9H),1.26(d,J=6.0Hz,3H).

[0497] MS (ESI) m / z 321.00 (M+H) + .

[0498] (2)(R)-N 2 Preparation of -(1-(6-(trifluoromethyl)pyridazine-3-amino)propan-2-yl)-1,3,4-thiadiazole-2,5-diamine

[0499]

[0500] Dissolve (R)-tert-butyl (1-((6-(trifluoromethyl)pyridazin-3-yl)amino)propan-2-yl)carbamate (1.00 g, 3.12 mmol) in 10 mL of dichloromethane, add 5 mL of trifluoroacetic acid dropwise in an ice-water bath, transfer to room temperature for 1 h after addition, remove the solvent by distillation under reduced pressure, dissolve in isopropanol solution, add 5-bromo-1,3,4-thiadiazole-2-amine (674 mg, 3.75 mmol) and N,N-diisopropylethylamine (484 mg, 3.75 mmol), mix well, transfer to 45°C and heat for 4 h, remove the solvent by distillation under reduced pressure, add water to quench the reaction, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Concentrate, separate by column chromatography, and the eluent is dichloromethane:methanol=10:1. 784 mg of off-white solid was obtained with a yield of 78.8% and a melting point of 75-76°C.

[0501] 1 H NMR (400MHz, Methanol-d4) δ7.57(d,J=9.7Hz,1H),7.04(d,J=9.4Hz,1H),4.00– 3.91(m,1H),3.78–3.69(m,1H),3.60–3.52(m,7.0Hz,1H),1.29(d,J=6.6Hz,3H).

[0502] MS (ESI) m / z 319.91 (M+H) + .

[0503] (3) Preparation of the title compound

[0504] (R)-N 2 -(1-(6-(trifluoromethyl)pyridazine-3-amino)propan-2-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.13 mmol), (S)-2-methoxy-2-phenylacetic acid (25 mg, 0.15 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 115 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, the reaction solution was moved to room temperature for 4 h, after which the reaction was quenched with water, extracted three times with ethyl acetate, washed twice with saturated sodium chloride solution, the organic phase was concentrated, and separated by column chromatography with the eluent being dichloromethane:methanol = 10:1 to obtain 36 mg of a white solid powder with a yield of 61.3% and a melting point of 121-123°C.

[0505] 1H NMR (400MHz, Methanol-d4) δ7.53(d,J=9.4Hz,1H),7.46(d,J=7.2Hz,2H),7.37(q,J=7.0Hz,3H),6.99(d, J=9.4Hz,1H),4.90(s,1H),4.08(q,J=6.5Hz,1H),3.79–3.54(m,2H),3.42(s,3H),1.30(d,J=6.8Hz,3H).

[0506] HRMS calcd.For C19H21F3N7O2S(M+H)+468.1351,found 468.1353.

[0507] Embodiment 46

[0508] (R)-2-(3-(trifluoromethoxy)phenyl)-N-(5-((1-(6-(trifluoromethyl)pyridazin-3-ylamino)propan-2-yl)amino)-1,3,4-thiadiazol-2-yl)acetamide

[0509]

[0510] (3) Preparation of the title compound

[0511] (R)-N 2 -(1-(6-(trifluoromethyl)pyridazine-3-amino)propan-2-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.13 mmol), 3-trifluoromethoxyphenylacetic acid (33 mg, 0.15 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (82 mg, 0.63 mmol) was added dropwise, the reaction solution was cooled to 0°C, 115 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, and the mixture was moved to room temperature for 4 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, and the organic phase was concentrated and separated by column chromatography. The eluent was dichloromethane:methanol = 10:1 to obtain 36 mg of white solid powder with a yield of 53.3% and a melting point of 119-121°C.

[0512] 1H NMR (400MHz, Methanol-d4) δ7.58(d,J=9.4Hz,1H),7.46(t,J=7.9Hz,1H),7.35(d,J=7.7Hz,1H),7.30(s,1H),7.23(d, J=8.3Hz,1H),7.05(d,J=9.4Hz,1H),4.11(q,J=6.4Hz,1H),3.84(s,2H),3.77–3.59(m,2H),1.35(d,J=6.6Hz,3H).HRMS calcd.ForC19H18F6N7O2S(M+H)+522.1068,found 522.1069.

[0513] Embodiment 47

[0514] (S)-2-methoxy-N-(5-((3R,5S)-5-methyl-1-(pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazole-2-

[0515] 2-Phenylacetamide

[0516]

[0517] (1) Preparation of tert-butyl (3R,5S)-1-(6-chloropyridazin-3-yl)-5-methylpyrrolidin-3-ylcarbamate

[0518]

[0519] 3,6-dichloropyridazine (1.00 g, 6.71 mmol), (3R, 5S)-5-methylpyrrolidin-3-ylcarbamic acid tert-butyl ester (1.61 g, 8.06 mmol) and N,N-diisopropylethylamine (1.04 g, 8.06 mmol) were added to the bottle in sequence, dissolved in 10 mL of acetonitrile, and heated to 80 ° C for 5 h. The reaction was cooled to room temperature, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic layer was washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentration and column chromatography separation were performed, and the eluent was petroleum ether: ethyl acetate = 1:1 to obtain 1.69 g of white solid, with a yield of 81.2% and a melting point of 130-131 ° C.

[0520] 1H NMR(400MHz,Chloroform-d)δ7.20(d,J=9.4Hz,1H),6.67(d,J=9.4Hz,1H),4.80(s,1H),4.25(s,2H),3.86(dd,J=11.0,6.6Hz,1 H),3.43(dd,J=11.0,5.4Hz,1H),2.56(dt,J=14.8,7.3Hz,1H),1.75(dt,J=12.5,5.5Hz,1H),1.46(s,9H),1.40(d,J=6.1Hz,3H).

[0521] MS (ESI) m / z 313.01 (M+H) + .

[0522] (2) Preparation of tert-butyl (3R,5S)-1-(pyridazin-3-yl)-5-methylpyrrolidin-3-ylcarbamate

[0523]

[0524] Dissolve (R)-tert-butyl (1-((6-chloropyridazin-3-yl)amino)propan-2-yl)carbamate (1.00 g, 3.20 mmol), ammonium formate (2.01 g, 32.00 mmol) and palladium / carbon (100 mg, 10%) in 10 mL of ethanol and react at room temperature for 3 h. Filter the insoluble solids through diatomaceous earth and distill the filtrate under reduced pressure to obtain 880 mg of a white solid with a yield of 98.9% and a melting point of 88-89°C.

[0525] 1 H NMR(400MHz,Chloroform-d)δ8.47(d,J=4.4Hz,1H),7.15(dd,J=9.5,4.5Hz,1H),6.63(d,J=9.0Hz,1H),5.08(s,1H),4.18(s,1H),3.83(t, J=8.9Hz,1H),3.39(dd,J=11.0,5.5Hz,1H),2.49(dt,J=14.4,7.6Hz,1H),1.72(dd,J=12.8,6.3Hz,1H),1.40(s,9H),1.33(d,J=6.3Hz,3H).

[0526] MS (ESI) m / z 279.04 (M+H) + .

[0527] (3)(3R,5S)-N 2 Preparation of 5-methyl-1-pyridazine-3-pyrrolidin-3-yl-1,3,4-thiadiazole-2,5-diamine

[0528]

[0529] Dissolve (3R,5S)-1-(pyridazine-3-yl)-5-methylpyrrolidin-3-ylcarbamic acid tert-butyl ester (1.00g, 3.59mmol) in 10mL dichloromethane, add 5mL trifluoroacetic acid dropwise under ice-water bath, transfer to room temperature for 1h after addition, remove the solvent by distillation under reduced pressure, dissolve in isopropanol solution, add 5-bromo-1,3,4-thiadiazole-2-amine (775mg, 4.31mmol) and N,N-diisopropylethylamine (557mg, 4.31mmol), mix well, transfer to 45℃ and heat for 4h, remove the solvent by distillation under reduced pressure, add water to quench the reaction, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Concentrate, separate by column chromatography, and the eluent is dichloromethane:methanol=10:1. 702 mg of off-white solid was obtained with a yield of 70.6% and a melting point of 110-112°C.

[0530] 1 H NMR (400MHz, DMSO-d6) δ8.48(s,1H),7.32(dd,J=9.3,4.6Hz,1H),7.13(s,1H),6.89(d,J=9.1Hz,1H),6.32(s,2H ),4.16(s,2H),3.86(t,J=9.4Hz,1H),3.47–3.40(m,1H),1.87–1.78(m,1H),1.30(d,J=5.9Hz,3H),1.26(s,1H).

[0531] MS (ESI) m / z 277.99 (M+H) + .

[0532] (4) Preparation of the title compound

[0533] (3R,5S)-N 2-5-methyl-1-pyridazine-3-pyrrolidin-3-yl-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.14 mmol), (S)-2-methoxy-2-phenylacetic acid (29 mg, 0.17 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (74 mg, 0.57 mmol) was added dropwise, the reaction solution was cooled to 0°C, 125 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, the reaction was moved to room temperature for 4 hours, after the reaction was completed, water was added to quench the reaction, ethyl acetate was extracted three times, saturated sodium chloride solution was washed twice, the organic phase was concentrated, and column chromatography was used for separation. The eluent was dichloromethane: methanol = 10:1, and 34 mg of white solid powder was obtained. The yield was 56.3%, and the melting point was 132-133°C.

[0534] 1 H NMR (400MHz, Methanol-d4) δ8.42(s,1H),7.46(s,2H),7.41–7.32(m,4H),6.98(dd,J=8.8,3.4Hz,1H),6.98(d,J=8.8Hz,1H),4.92(d,J=3.4Hz,1H ),4.35(s,1H),4.26(s,1H),4.04–3.96(m,1H),3.56(d,J=10.8Hz,1H),3 .42(d,J=3.4Hz,3H),2.60(s,1H),1.99–1.91(m,1H),1.39–1.35(m,3H).

[0535] HRMS calcd.For C20H24N7O2S(M+H)+426.1632,found 426.1629.

[0536] Embodiment 48

[0537] (S)-2-methoxy-2-phenyl-N-(5-((R)-1-(pyridazin-3-yl)piperidin-3-amino)-1,3,4-thiadiazol-2-yl)

[0538] Acetamide

[0539]

[0540] (1) Preparation of tert-butyl (R)-(1-(6-chloropyridazin-3-yl)piperidin-3-yl)carbamate

[0541]

[0542] 3,6-dichloropyridazine (1.00 g, 6.71 mmol), (R)-piperidin-3-ylcarbamic acid tert-butyl ester (1.61 g, 8.06 mmol) and N,N-diisopropylethylamine (1.04 g, 8.06 mmol) were added to the bottle in sequence, dissolved in 10 mL of acetonitrile, and heated to 80 ° C for 5 h. The reaction was cooled to room temperature, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic layer was washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentration and column chromatography were performed, and the eluent was petroleum ether: ethyl acetate = 1:1 to obtain 1.66 g of white solid, with a yield of 79.3% and a melting point of 156-157 ° C.

[0543] 1 H NMR (400MHz, Chloroform-d) δ7.22(d,J=9.7Hz,1H),7.03(d,J=9.7Hz,1H),4.68(s,1H),4.00(d,J=11.1Hz,1H),3.88(d,J=11.4H z,1H),3.71(s,1H),3.54(s,1H),3.36(s,1H),2.08–1.96(m,1H),1.89–1.80(m,1H),1.76–1.68(m,1H),1.64(s,1H),1.47(s,9H).

[0544] MS (ESI) m / z 312.97 (M+H) + .

[0545] (2) Preparation of tert-butyl (R)-(1-(pyridazin-3-yl)piperidin-3-yl)carbamate

[0546]

[0547] Dissolve (R)-tert-butyl (1-(6-chloropyridazin-3-yl)piperidin-3-yl)carbamate (1.00 g, 3.20 mmol), ammonium formate (2.01 g, 32.00 mmol) and palladium / carbon (100 mg, 10%) in 10 mL of ethanol and react at room temperature for 3 h. Remove the insoluble solid by filtration through diatomaceous earth, and distill the filtrate under reduced pressure to obtain 870 mg of a white solid with a yield of 97.7% and a melting point of 85-86°C.

[0548] 1H NMR(400MHz,Chloroform-d)δ8.56(s,1H),7.23–7.14(m,1H),6.99(d,J=9.2Hz,1H),4.69(s,1H),3.95(s,1H),3.86(d,J= 12.6Hz,1H),3.70(s,1H),3.52(s,1H),3.35(s,1H),1.97(s,1H),1.80(s,1H),1.74–1.54(m,2H),1.44(s,9H).MS(ESI)m / z 279.04(M+H) + .

[0549] (3)(R)-N 2 Preparation of -(1-pyridazine-3-yl)pyridin-3-yl)-1,3,4-thiadiazole-2,5-diamine

[0550]

[0551] Dissolve (R)-(1-(pyridazine-3-yl)piperidin-3-yl)carbamic acid tert-butyl ester (1.00g, 3.59mmol) in 10mL dichloromethane, add 5mL trifluoroacetic acid dropwise in an ice-water bath, transfer to room temperature for reaction for 1h after addition, remove the solvent by distillation under reduced pressure, dissolve in isopropanol solution, add 5-bromo-1,3,4-thiadiazole-2-amine (775mg, 4.31mmol) and N,N-diisopropylethylamine (557mg, 4.31mmol), mix well, transfer to 45℃ and heat for reaction for 4h, remove the solvent by distillation under reduced pressure, add water to quench the reaction, extract the mixture three times with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. Concentrate, separate by column chromatography, and the eluent is dichloromethane:methanol=10:1. Obtain 731mg of off-white solid, yield 73.5%, melting point 120-121℃.

[0552] 1 H NMR (400MHz, DMSO-d6) δ8.50(s,1H),7.33(dd,J=9.3,4.7Hz,1H),7.18(d,J=9.3Hz,1H),6.92(d,J=6.7Hz,1H),6.31(s,2H),4.2 6(d,J=12.8Hz,1H),4.07–3.93(m,1H),3.56(s,1H),3.25(s,1H),3.07(t,J=10.5Hz,1H),1.99(s,1H),1.78(s,1H),1.54(s,2H).

[0553] MS (ESI) m / z 277.97 (M+H) + .

[0554] (4) Preparation of the title compound

[0555] (R)-N 2 -(1-pyridazine-3-yl)pyridin-3-yl)-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.14 mmol), (S)-2-methoxy-2-phenylacetic acid (29 mg, 0.17 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (74 mg, 0.57 mmol) was added dropwise, the reaction solution was cooled to 0°C, 125 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, the reaction mixture was moved to room temperature for 4 h, after which the reaction was quenched with water, the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, the organic phase was concentrated, and the mixture was separated by column chromatography with the eluent being dichloromethane:methanol = 10:1 to give 38 mg of a white solid powder with a yield of 64.4% and a melting point of 124-125°C.

[0556] 1 H NMR (400MHz, Methanol-d4) δ8.41(d,J=4.3Hz,1H),7.46(d,J=6.7Hz,2H),7.41–7.29(m,4H),7.23(d,J=9.3Hz,1H),4.91(s,1H),4.15(d,J=13 .2Hz,1H),3.94(d,J=13.2Hz,1H),3.83(s,1H),3.48(s,1H),3.44–3.4 0(m,3H),3.35(s,1H),2.11(s,1H),1.89(s,1H),1.69(d,J=7.5Hz,2H).

[0557] HRMS calcd.For C20H24N7O2S(M+H)+426.1632,found 426.1631.

[0558] Embodiment 49

[0559] (5-(((3S,4S)-4-Fluoro-1-pyridazin-3-yl)pyrrolidin-3-amino)-1,3,4-thiadiazol-2-yl)-2-methoxy-2-phenylacetamide

[0560]

[0561] (1) Preparation of tert-butyl (3S,4S)-1(-(6-chloropyridazin-3-yl)-4-fluoropyrrolidin-3-yl)carbamate

[0562]

[0563] 3,6-dichloropyridazine (1.00 g, 6.71 mmol), (3S,4S)-4-fluoropyrrolidin-3-ylcarbamic acid tert-butyl ester (1.65 g, 8.06 mmol) and N,N-diisopropylethylamine (1.04 g, 8.06 mmol) were added to the bottle in sequence, dissolved in 10 mL of acetonitrile, and heated to 80 ° C for 5 h. The reaction was cooled to room temperature, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic layer was washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentration and column chromatography were performed, and the eluent was petroleum ether: ethyl acetate = 1:1 to obtain 1.71 g of white solid, with a yield of 80.4% and a melting point of 146-147 ° C.

[0564] 1 H NMR (400MHz, Chloroform-d) δ7.16(d,J=9.3Hz,1H),6.59(d,J=9.3Hz,1H),5.28(s,1H),5.16(s,1H),4.44(s,1H),3.99–3.58(m,4H),1.44(s,9H).

[0565] MS (ESI) m / z 316.94 (M+H) + .

[0566] (2) Preparation of tert-butyl (3S,4S)-1-((pyridazine-3-yl)-4-fluoropyrrolidin-3-yl)carbamate

[0567]

[0568] Dissolve tert-butyl (3S,4S)-1-((6-chloropyridazin-3-yl)-4-fluoropyrrolidin-3-yl)carbamate (1.00 g, 3.16 mmol), ammonium formate (2.04 g, 32.41 mmol) and palladium / carbon (100 mg, 10%) in 10 mL of ethanol and react at room temperature for 3 h. Filter through diatomaceous earth to remove insoluble solids, and distill the filtrate under reduced pressure to obtain 876 mg of colorless oil with a yield of 98.3%.

[0569] 1 H NMR(400MHz,Chloroform-d)δ8.47(s,1H),7.17–7.12(m,1H),6.56(d,J=9.1Hz,1H) ,5.30–5.10(m,1H),4.40(s,1H),3.93–3.76(m,2H),3.72–3.60(m,1H),1.42(s,9H).

[0570] MS (ESI) m / z 283.02 (M+H) + .

[0571] (3)N 2 Preparation of -(3S,4S)-4-fluoro-1-pyridazin-3-ylpyrrolidine-1,3,4-thiadiazole-2,5-diamine

[0572]

[0573] Dissolve tert-butyl ((3S,4S)-1-((pyridazine-3-yl)-4-fluoropyrrolidin-3-yl)carbamate (1.00 g, 3.54 mmol) in 10 mL of dichloromethane, add 5 mL of trifluoroacetic acid dropwise under ice-water bath, transfer to room temperature for reaction for 1 h, remove the solvent by distillation under reduced pressure, dissolve in isopropanol solution, add 5-bromo-1,3,4-thiadiazole-2-amine (764 mg, 4.25 mmol) and N,N- Diisopropylethylamine (549 mg, 4.25 mmol) was mixed evenly and then transferred to 45°C for heating and reaction for 4 h. The reaction solution was distilled under reduced pressure to remove the solvent, and water was added to quench the reaction. The mixture was extracted with ethyl acetate three times, and the organic layer was washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentration and column chromatography were performed, and the eluent was dichloromethane: methanol = 10:1. 782 mg of off-white solid was obtained, with a yield of 78.6% and a melting point of 122-123°C.

[0574] 1 H NMR(400MHz, DMSO-d6)δ8.54(d,J=4.5Hz,1H),7.42–7.37(m,1H),7.22(s,1H),6.95( d,J=8.9Hz,1H),6.43(s,2H),5.36(d,J=50.9Hz,1H),4.37(s,1H),3.98–3.56(m,4H).

[0575] MS (ESI) m / z 281.92 (M+H) + .

[0576] (4) Preparation of the title compound

[0577] N 2-(3S,4S)-4-Fluoro-1-pyridazin-3-ylpyrrolidine-1,3,4-thiadiazole-2,5-diamine (40 mg, 0.14 mmol), (S)-2-methoxy-2-phenylacetic acid (29 mg, 0.17 mmol) were dissolved in 1 mL of N,N-dimethylformamide, N,N-diisopropylethylamine (74 mg, 0.57 mmol) was added dropwise, the reaction solution was cooled to 0°C, 125 μL of 50% propylphosphonic anhydride in ethyl acetate was added dropwise, the reaction mixture was moved to room temperature for 4 h, after which the reaction was quenched with water, the mixture was extracted with ethyl acetate three times, washed with saturated sodium chloride solution twice, the organic phase was concentrated, and the mixture was separated by column chromatography with the eluent being dichloromethane:methanol = 10:1 to give 37 mg of a white solid powder with a yield of 61.0% and a melting point of 137-138°C.

[0578] 1 H NMR(400MHz, Methanol-d4)δ8.47(d,J=5.1Hz,1H),7.46(s,2H),7.42–7.34(m,4H),7.04–6.98(m,1H),5. 37(d,J=50.2Hz,1H),4.92(d,J=4.2Hz,1H),4.57(s,1H),3.90(s,2H),3.80(s,2H),3.42(d,J=4.0Hz,3H).

[0579] HRMS calcd.For C19H21FN7O2S(M+H)+430.1359,found 430.1360.

[0580] Pharmacological experiments

[0581] Experimental Example 1: Screening of GAC inhibitor enzymatic inhibition level

[0582] Experimental purpose: To determine the enzyme activity of GAC inhibitors using a glutaminase activity detection kit, and to evaluate the inhibitory activity and specificity of the compounds of the present invention against GAC protein in vitro.

[0583] Experimental method: The fragment encoding GAC protein (NP_001106854.1, amino acids 73 to 598) was cloned into the pET28a plasmid vector and transformed into E. coli (DE3), and protein expression was induced and purified to obtain high-purity GAC protein. After incubating 10 nM GAC protein with the test compound at room temperature for 10 minutes, the GAC enzyme activity was detected using a glutaminase activity detection kit (abcam, ab284547) to calculate the inhibitory effect of the compound on GAC enzyme activity. The EC value of the compound inhibiting GAC enzyme activity was fitted using Graphpad 50 .

[0584] Experimental results: As shown in Table 1, the compounds of the present invention have high GAC protein inhibitory activity, most of the EC 50 The values ​​are all at nM level, indicating that the compounds of the present invention directly act on GAC protein and have a strong inhibitory effect, and their effects are comparable to those of the disclosed CB839.

[0585] Table 1 Inhibitory effect of the compounds of the present invention on GAC protein

[0586]

[0587]

[0588] *A<100nM; 100nM≤B<500nM; 500nM≤C<1000nM; D≥1000nM

[0589] ND Not determined

[0590] Experimental Example 2: Detection of Anti-tumor Cell Proliferation Activity of Compounds

[0591] Experimental purpose: To evaluate the anti-tumor activity of the compounds of the present invention at the cellular level by measuring the inhibition of A549 cell proliferation by GAC inhibitors.

[0592] Experimental method: Glutamine-dependent A549 cells were seeded into 96-well plates at a density of 1,500 cells / well. The next day, different concentrations of compound-containing culture medium were replaced and the cells were cultured for 72 hours. Finally, the cell culture plate was restored to room temperature, and Cell Titer-Glo (Promega, G7570) reagent was added. After shaking, the chemiluminescence value was detected using a BioTek Synergy H1 microplate reader. Graphpad was used to fit the IC values ​​of the compounds for inhibiting tumor proliferation. 50 .

[0593] Experimental results: As shown in Table 2, the compounds of the present invention have high GAC protein inhibitory activity, most of the IC 50 The values ​​are all at nM level, indicating that the compounds of the present invention directly act on GAC protein and have a strong inhibitory effect, and their effects are comparable to those of the disclosed CB839.

[0594] Table 2 Inhibitory effect of the compounds of the present invention on GAC protein

[0595]

[0596]

[0597] *A<100nM; 100nM≤B<500nM; 500nM≤C<1000nM; D≥1000nM

[0598] ND Not determined

[0599] Experimental Example 3: A549 nude mouse xenograft tumor experiment

[0600] Experimental purpose: To study the inhibitory effect of the compound on solid tumors by oral administration and to evaluate the in vivo anti-tumor activity of the compound of the present invention.

[0601] Experimental method: A549 tumor tissue was removed from tumor-bearing mice under sterile conditions, cut into 1mm×1mm tumor pieces, and evenly inoculated into the subcutaneous tissue of the axilla of male Balb / c nude mice. After 7 days, when the tumor grew to 100mm 3 After that, the animals were randomly divided into groups and the administration began. The experiment set up a solvent control group, a CB839 100 mg / kg group, an Example 22 100 mg / kg group, an Example 22 150 mg / kg group, an Example 35 100 mg / kg group, and an Example 35 150 mg / kg group, all of which were orally administered once a day. The body weight was weighed twice a week and the length and width of the tumor were measured with a vernier caliper. After continuous administration for 24 days, the tumor inhibition rate was calculated to evaluate the intensity of the anti-tumor effect.

[0602] Experimental results: Figure 1 and Figure 2 As shown, the tumor inhibition rates of Example 22 and Example 35 at oral doses of 100 mg / kg and 150 mg / kg were higher than that of 100 mg / kg of CB839, and the tumor volume showed significant differences compared with the blank group, indicating that Example 22 and Example 35 have good in vivo anti-tumor activity.

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof: in, R1 is selected from H, F, Cl, Br, I, CF3; R2 is selected from H, OCH3, C 1-6 alkyl; R3 is selected from H; Or R2 and R3 are connected together to form a C 3-6 Cycloalkyl; Ar is selected from substituted or unsubstituted aromatic rings or aromatic heterocycles or phenylmethyl; the aromatic rings or aromatic heterocycles are selected from five-membered rings, six-membered rings or naphthalene rings; the substituents are selected from C 1-3 Alkyl, C 1-3 Alkoxy, halogen, halogen-substituted alkyl, halogen-substituted alkoxy, hydroxy, hydroxymethyl, cyano, phenyl, phenoxy, phenyl-substituted alkyl, carboxyl, carboxylate, amide, alkyl-substituted or halogen-substituted alkyl-substituted amide, phosphate, phosphate, boric acid, borate, tetrazolyl and alkylsulfonylcarbonyl; the aromatic heterocycle contains 1-2 heteroatoms selected from N, O and S; Ring L is selected from substituents having the following structure:

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that Ar is selected from unsubstituted or substituted methyl, hydroxyl, hydroxymethyl, methoxy, trifluoromethyl, trifluoromethoxy, F, Cl, Br, cyano, carboxyl, phosphate, borate, tetrazolyl, An aromatic ring or aromatic heterocyclic group substituted with any one or more substituents; the aromatic ring group is selected from phenyl and naphthyl; the aromatic heterocyclic group is selected from pyridyl, pyrrolyl, furyl and thienyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that The pharmaceutically acceptable salt is selected from organic acid salts or inorganic acid salts, including hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, malate, fumarate, tartrate or methanesulfonate.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that The compound is selected from:

5. A method for preparing the compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Here are the steps: Route (1): R=Cl or CF3 Wherein R1, R2, R3, Ar are defined as described in any one of claims 1-4. Reagents and conditions: (a) N-tert-butoxycarbonyl substituted diamine, N,N-diisopropylethylamine, acetonitrile, heating to reflux; (b) ammonium formate, palladium / carbon, ethanol; (c) trifluoroacetic acid, dichloromethane; (d) 5-bromo-1,3,4-thiadiazol-2-amine, N,N-diisopropylethylamine, isopropanol, 45°C; (e) unsubstituted phenylpropionic acid or substituted aromatic acetic acid, propylphosphonic anhydride, N,N-diisopropylethylamine, N,N-dimethylformamide.

6. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

7. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier.

8. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof or the composition according to claim 7 in the preparation of GAC inhibitor-related drugs.

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