Substituted oxadiazole compounds, preparation methods and uses thereof

Abstract

The present invention belongs to the field of pharmaceutical chemistry technology, and specifically relates to a series of substituted oxadiazole compounds, preparation methods and uses thereof. Pharmacological experiments show that the series of compounds have strong selectivity and inhibitory effects on SphK2, and have anti-proliferation effects on cervical cancer cells, prostate cancer cells and colon cancer cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical chemistry, and relates to a series of substituted oxadiazole compounds, preparation methods and uses thereof, and specifically includes substituted oxadiazole compounds and pharmaceutically acceptable salts thereof, preparation methods thereof, pharmaceutical compositions and uses thereof. Background Art

[0002] Cancer is a disease caused by the uncontrolled proliferation and spread of abnormal cells. There are many targets related to cancer, among which the sphingosine kinase (SphKs) gene has the properties of proto-oncogenes and is an ideal target for cancer treatment. There are two subtypes of SphKs protein: SphK1 and SphK2, which are widely distributed in mammals, but their tissue distribution and physiological functions are different. Current studies have found that SphK1 is highly expressed in a variety of cancer cells, including colon cancer, glioma, rectal cancer, gastric cancer, lung cancer, kidney cancer, ovarian cancer, breast cancer, and cervical cancer. Compared with SphK1, SphK2 is localized in the nucleus, can inhibit DNA synthesis and regulate the activity of histone deacetylase (HDAC), and is highly expressed in inflammation, central nervous system diseases, and tumors of the rectum, stomach, kidney, colon, uterus, ovary, breast, and lung. The ligand (Sph) binding regions of the two subtypes of SphKs are very similar but also have important differences, which makes SphK1 and SphK2 have different sensitivities to inhibitors. The crystal structure of human SphK1 protein was solved in 2013. It contains 384 amino acid residues, including 9 α helices and 17 β folds. N -Terminal domain ( N -terminaldomain, NTD) and C -Terminal domain ( C -terminaldomain, CTD), contains the ATP catalytic site in the middle of the two domains. The two helices α7 / α8 at the CTD end control the entry and exit of the substrate Sph. Studies have found that Sph can form strong hydrogen bond interactions with the amino acid residues Asp81, Ser168 and Asp178 in SphK1. At present, most SphKs inhibitors inhibit SphKs by competing for the Sph substrate binding site, and very few exert their inhibitory effects by competing for the ATP binding site.

[0003] K145, chemically named 3-(2-amino-ethyl)-5-[3-(4-butoxy-phenyl)-propylidene]-thiazolidine-2, 4-dione, is one of the most representative SphK2 inhibitors. Its IC 50 The value was 4.30 μM, and its effect on SphK2 KiThe concentration of ABC294640 is 6.40 μM, and it is often used as a tool drug. Currently, there are few highly effective and selective SphK2 inhibitors reported, and ABC294640 is the only known SphK2 selective inhibitor that has entered Phase II clinical trials.

[0004] As the incidence of cancer and inflammatory diseases is increasing year by year and their pathological mechanisms are complex, it is urgent to find SphK2 inhibitors with diverse structures to alleviate cancer and inflammatory diseases. Based on the chemical structure of K145, the researchers discovered substituted oxadiazole compounds, which have good selectivity for SphK2 and anti-tumor cell proliferation activity. Summary of the invention

[0005] The technical problem solved by the present invention is to provide a series of substituted oxadiazole compounds and pharmaceutically acceptable salts thereof, preparation methods and pharmaceutical compositions thereof, and use thereof in the preparation of drugs for preventing and / or treating diseases mediated by SphK2.

[0006] In order to solve the technical problem of the present invention, the present invention provides the following technical solutions:

[0007] 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:

[0008]

[0009] Formula (I)

[0010] Among them, R is

[0011] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0012] Preferably, in the above compound or its pharmaceutically acceptable salt or optical isomer thereof, R is

[0013] , , , , , , , , , , More preferably, in the above compound or its pharmaceutically acceptable salt or optical isomer thereof, R is , , , .

[0014] More preferably, the R is , , .

[0015] The pharmaceutically acceptable salt of any of the above compounds of the present invention is an organic acid salt or an inorganic acid salt, wherein the organic acid includes acetic acid, trifluoroacetic acid, methanesulfonic acid, toluenesulfonic acid, maleic acid, succinic acid, tartaric acid, citric acid, and fumaric acid; the inorganic acid includes hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and phosphoric acid.

[0016] The second aspect of the technical solution of the present invention is to provide a method for preparing the compound or a pharmaceutically acceptable salt thereof according to the first aspect. The compound of the general formula (I) of the present invention can be prepared by the following method:

[0017] The intermediate compound 14-46 (3 mmol, 1 equiv) and trifluoroacetic acid (3 mL) were dissolved in dichloromethane (6 mL). After stirring at room temperature for 1 hour, TLC was detected (dichloromethane: methanol = 7:1). After the reaction was completed, the reaction solution was dried, and the crude product was separated by silica gel column chromatography (dichloromethane: methanol = 15:1) to obtain series d white solid compounds with a yield of 85-95%.

[0018] Compound 14-46 was synthesized from 1 as raw material through three steps of reaction:

[0019]

[0020] Among them, R is

[0021] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0022] The structural formula of intermediate compound 14-46 is as follows:

[0023]

[0024] The third aspect of the technical solution of the present invention is to provide a pharmaceutical composition comprising the compound described in the first aspect, its pharmaceutically acceptable salt, its optical isomer and one or more pharmaceutical carriers and / or diluents, in any clinically or pharmaceutically acceptable dosage form, preferably an oral preparation or an injection. It contains a physiologically effective amount of 0.01g to 10g of the compound represented by the general formula (I), which can be 0.01g, 0.015g, 0.02g, 0.025g, 0.03g, 0.04g, 0.05g, 0.1g, 0.125g, 0.2g, 0.25g, 0.3g, 0.4g, 0.5g, 0.6g, 0.75g, 1g, 1.25g, 1.5g, 1.75g, 2g, 2.5g, 3g, 4g, 5g, 6g, 7g, 8g, 9g, 10g, etc.

[0025] Any compound of the present invention or a pharmaceutically acceptable salt thereof can be administered orally or parenterally to a patient in need of such treatment.

[0026] When used for parenteral administration, it can be made into an injection. When it is made into an injection, it can be produced by conventional methods in the existing pharmaceutical field, and aqueous solvents or non-aqueous solvents can be selected. The most commonly used aqueous solvent is water for injection, and 0.9% sodium chloride solution or other suitable aqueous solutions can also be used; the commonly used non-aqueous solvent is vegetable oil, mainly soybean oil for injection, and other aqueous solutions of ethanol, propylene glycol, polyethylene glycol, etc. When preparing an injection, no additives may be added, or appropriate additives may be added according to the properties of the drug, such as osmotic pressure regulators, pH regulators, solubilizers, fillers, antioxidants, antibacterial agents, emulsifiers, suspending agents, etc. When used for oral administration, it can be made into conventional solid preparations, such as tablets, capsules, pills, granules, etc.; it can also be made into oral liquid preparations, such as oral solutions, oral suspensions, syrups, etc. When making oral preparations, appropriate fillers, binders, disintegrants, lubricants, etc. may be added.

[0027] The compounds of the present invention can be added with pharmaceutically acceptable carriers to prepare common pharmaceutical preparations, such as tablets, capsules, powders, syrups, liquids, suspensions, injections, and can be added with common pharmaceutical excipients such as flavors, sweeteners, liquid or solid fillers or diluents.

[0028] The compounds of the present invention can be administered clinically by oral administration, injection, or the like.

[0029] The clinical dosage of the compound of the present invention is 0.01-1000 mg / day, and may deviate from this range depending on the severity of the disease or the dosage form.

[0030] The fourth aspect of the technical solution of the present invention is to provide use of the compound described in the first aspect and the pharmaceutical composition described in the third aspect in the preparation of a drug for preventing and / or treating a disease mediated by SphK2.

[0031] The SphK2-mediated diseases include cancer, inflammatory diseases, ischemia-reperfusion injury, renal fibrosis, Alzheimer's disease, diabetic retinopathy, and diabetic nephropathy.

[0032] Preferably, the cancer includes colorectal cancer, gastric cancer, kidney cancer, colon cancer, uterine cancer, ovarian cancer, breast cancer and lung cancer.

[0033] Preferably, the inflammatory disease includes ulcerative colitis and rheumatoid arthritis.

[0034] Beneficial technical effects:

[0035] (1) The compounds of the present invention have strong selectivity and inhibitory effects on SphK2. Such compounds and their pharmaceutical preparations can be used to treat a series of diseases mediated by SphK2, such as cervical cancer, prostate cancer, colon cancer, inflammatory bowel disease, renal fibrosis, etc.

[0036] (2) The preparation method provided by the present invention has the characteristics of mild reaction conditions and simple operation. DETAILED DESCRIPTION

[0037] The present invention will be further described below in conjunction with specific implementation modes so that those skilled in the art can better understand the present invention, but the present invention is not limited thereto.

[0038] Compound 14-46 was synthesized from 1 as raw material through three steps of reaction.

[0039]

[0040] The raw material 1 (10 mmol, 1 equiv), hydroxylamine hydrochloride (11 mmol, 1.1 equiv), and sodium bicarbonate (11 mmol, 1.1 equiv) were dissolved in methanol (50 mL). After reflux reaction for 22 hours, TLC detection (developing solvent: dichloromethane: methanol = 10:1) was performed. After the reaction was completed, the reaction solution was filtered and the filtrate was spin-dried, extracted with water and ethyl acetate, and the organic phase was washed with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation to obtain a white solid compound 2 with a yield of 95%.

[0041] Compound 2 (8 mmol, 1 equiv), 2-(1-(tert-butyloxycarbonyl)azetidin-3-yl)acetic acid (8.8 mmol, 1.1 equiv), and DIPEA (8.8 mmol, 1.1 equiv) were dissolved in DMF (40 mL), and then HATU (8.8 mmol, 1.1 equiv) was added. o After heating and stirring at 4 °C for 4 hours, TLC was performed (developing solvent: n-hexane: ethyl acetate = 10:1). After the reaction was completed, the reaction solution was filtered and the filtrate was dried by spin drying, extracted with water and ethyl acetate, the organic phase was washed with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was separated by silica gel column chromatography (mobile phase: n-hexane: ethyl acetate = 25:1) to obtain white solid compound 3 with a yield of 90%. Compound 3 (6 mmol, 1 equiv), cesium carbonate (6.6 mmol, 1.1 equiv), and p-trifluoromethylbenzenesulfonyl chloride (6.6 mmol, 1.1 equiv) were dissolved in DMF (30 mL). 65 o After heating and stirring at C for 12 hours, TLC was performed (n-hexane: ethyl acetate = 15:1). After the reaction was completed, the reaction solution was extracted with water and ethyl acetate, and the organic phase was washed with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain a crude product 14-46.

[0042] The series of compounds of the present invention can be prepared by the following method:

[0043]

[0044] The target compounds d1-d33 were obtained by one-step reaction of the intermediate compounds 14-46. The intermediate compounds 14-46 (3 mmol, 1 equiv) and trifluoroacetic acid (3 mL) were dissolved in dichloromethane (6 mL). After stirring at room temperature for 1 hour, TLC was detected (dichloromethane: methanol = 7:1). After the reaction was completed, the reaction solution was dried, and the crude product was separated by silica gel column chromatography (dichloromethane: methanol = 15:1) to obtain white solid compounds d1-d33 with a yield of 85-95%.

[0045] Example 1: 4-(5-(azetidin-3-ylmethyl)-3-(4-(4-(trifluoromethyl)benzenesulfonic acid)phenyl)-1,2,4-oxadiazole trifluoroacetate (d1):

[0046]

[0047] The intermediate compound used in the synthesis is 14. White solid powder, yield 91%, 1 HNMR (800 MHz, Methanol- d 4 ) d 8.09–8.07 (m, 2H), 8.05 (d, J =7.9 Hz, 2H), 7.96 (d, J =8.1 Hz, 2H), 7.21 (d, J =7.9 Hz, 2H), 4.28 (t, J =9.7 Hz, 2H), 4.10–4.06 (m, 2H), 3.38 (d, J =7.6 Hz,2H), 3.31 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 167.06, 151.46, 129.20,128.79,126.47,125.96,122.69,50.67,28.49. IR(KBr,cm -1):2978, 2671, 1674, 1581,1411, 1384, 1329, 1208, 1181, 1137, 869. HRMS(ESI): m / z Calculate for C 19 H 16 F 3 N 3 O 4 S[M+H] + : 440.0814. found: 440.0814.

[0048] Example 2: 4-(5-(azetidin-3-ylmethyl)-3-(4-(4-bromobenzenesulfonic acid)phenyl)-1,2,4-oxadiazole trifluoroacetate (d2):

[0049]

[0050] The intermediate compound used in the synthesis is 15. White solid powder, yield 90%, 1 HNMR (800 MHz, Methanol- d 4 ) d 8.04(d, J =7.8 Hz, 2H),7.81 (d, J =7.8 Hz, 2H), 7.76 (d, J =8.0 Hz,2H), 7.19(d, J =7.9 Hz,2H), 4.28 (t, J =9.7 Hz, 2H), 4.10–4.05 (m, 2H), 3.38 (d, J =7.5 Hz,2H), 3.35 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 177.64, 167.12, 151.60,134.12, 132.65, 129.95, 129.59, 128.74, 122.77, 50.69, 28.49. IR (KBr, cm -1 ):2967, 1674, 1575, 1384, 1203, 1181, 1164, 869, 770, 721. HRMS(ESI): m / z calcdfor C 18 H16 Bn 3 O 4 S [M+H] + : 450.0045. found: 450.0065.

[0051] Example 3: 4-(5-(azetidin-3-ylmethyl)-3-(4-(4-fluorobenzenesulfonic acid)phenyl)-1,2,4-oxadiazole trifluoroacetate (d3):

[0052]

[0053] The intermediate compound used in the synthesis is 16. The product is a white solid powder with a yield of 90%. 1 HNMR (800 MHz, Methanol- d 4 ) d 8.02 (d, J =7.5 Hz, 2H), 7.94–7.89 (m, 2H), 7.36 (t, J =8.0 Hz, 2H),7.17 (d, J =7.5Hz, 2H), 4.28 (t, J =9.6Hz, 2H), 4.11–4.05 (m, 2H), 3.38 (d, J =7.6Hz, 2H), 3.31 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 177.66, 167.10, 167.00,165.72, 151.62, 131.52, 131.47, 128.70, 122.77, 116.58, 116.47, 50.66, 28.51.IR (KBr, cm -1 ): 2676, 1685, 1384, 1206, 1178, 1160, 1096, 873, 839, 570, 551.HRMS(ESI): m / z Calculate for C 18 H 16 FN 3 O 4 S [M+H] + : 390.0846. found: 390.0859.

[0054] Example 4: 4-(5-(azetidin-3-ylmethyl)-3-(4-(3-fluorobenzenesulfonic acid)phenyl)-1,2,4-oxadiazole trifluoroacetate (d4):

[0055]

[0056] The intermediate compound used in the synthesis is 17. The product is a white solid powder with a yield of 91%. 1 HNMR (800 MHz, Methanol- d 4 ) d 8.04 (d, J =7.9Hz, 2H), 7.71–7.60 (m, 3H), 7.55 (t, J =8.3Hz, 1H),7.19 (d, J =7.9Hz, 2H), 4.27 (t, J =9.7Hz, 2H), 4.10–4.03 (m, 2H), 3.38 (d, J =7.5Hz, 2H), 3.31 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 161.79, 151.57, 136.86,131.54, 128.73, 125.88, 124.42, 122.70, 121.78, 121.67, 115.38, 115.25,50.69, 29.40. IR (KBr, cm -1 ): 2973, 1669, 1578, 1378, 1232, 1200, 1178, 1153,1139, 864, 769. HRMS (ESI): m / z Calculate for C 18 H 16 FN 3 O 4 S [M+H] + : 390.0846. found:390.0863.

[0057] Example 5: 4-(5-(azetidin-3-ylmethyl)-3-(4-(benzenesulfonic acid)phenyl)-1,2,4-oxadiazole trifluoroacetate (d5):

[0058]

[0059] The intermediate compound used in the synthesis is 18. The product is a white solid powder with a yield of 95%. 1 HNMR (800 MHz, Methanol- d 4 ) d 8.01 (d, J =7.9 Hz, 2H), 7.86 (d, J =7.9 Hz, 2H), 7.76 (t, J =7.5 Hz,1H), 7.62(t, J =7.6 Hz, 2H), 7.15 (d, J =8.0 Hz, 2H), 4.28 (t, J =9.7 Hz, 2H),4.10–4.05 (m, 2H), 3.37 (d, J =7.6 Hz, 2H). 13 CNMR(200 MHz, Methanol- d 4 ) d 177.61,167.13, 135.01, 134.55, 129.26, 128.61, 128.25, 125.64, 122.76, 50.68, 29.33,28.49. IR (KBr, cm -1 ): 2989, 1670, 1578, 1424, 1372, 1204, 1183, 1133, 863,774, 581, 565. HRMS (ESI): m / z Calculate for C 18 H 17 N 3 O 4 S [M+H] + : 372.0940. found:372.0960.

[0060] Example 6: 4-(5-(azetidin-3-ylmethyl)-3-(4-(3,4,5-trifluorobenzenesulfonic acid)phenyl)-1,2,4-oxadiazole trifluoroacetate (d6):

[0061]

[0062] The intermediate compound used in the synthesis is 19. The product is a white solid powder with a yield of 92%. 1 HNMR (800 MHz, Methanol-d 4 ) d 8.08 (d, J =7.4 Hz, 2H), 7.76 (t, J =5.8 Hz, 2H), 7.26 (d, J =7.4 Hz,2H), 4.28 (t, J =9.7 Hz, 2H), 4.11–4.06 (m, 2H), 3.39 (d, J =7.6 Hz, 2H), 1.43–1.13(m, 1H). 13 CNMR(200 MHz, Methanol- d 4 ) d 167.07, 151.80, 151.75, 151.37,150.53, 150.48, 128.89, 126.15, 122.72, 114.09, 114.07, 114.00, 113.97,50.68, 29.33, 28.51. IR (KBr, cm -1 ): 3071, 2666, 1669, 1582, 1528, 1385, 1331,1201, 1171, 1142, 874, 601. HRMS (ESI): m / z Calculate for C 18 H 14 F 3 N 3 O 4 S [M+H] + :426.0657. found: 426.0667.

[0063] Example 7: 4-(5-(azetidin-3-ylmethyl)-3-(4-(propyl-2-yn-1-yloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d7):

[0064]

[0065] The intermediate compound used in the synthesis is 20. The product is a white solid powder with a yield of 92%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.99 (d, J =7.6 Hz, 2H), 7.11 (d,J =7.7 Hz, 2H), 4.81 (s, 2H), 4.28(t, J =9.6 Hz, 2H), 4.11–4.05 (m, 2H), 3.37 (d, J =7.5Hz, 2H), 2.99 (s, 1H), 1.29(s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 177.02, 160.26, 128.53, 119.47, 115.02,77.89, 50.76, 29.41, 28.53. IR (KBr,cm -1 ): 3236, 2923, 1680, 1620, 1577, 1485,1429, 1356, 1247, 1211, 1177, 1124, 1024, 836. HRMS (ESI): m / z Calculate for C 15 H 15 N 3 O 2 [M+H] + : 270.1164. found: 270.1200.

[0066] Example 8: 4-(5-(azetidin-3-ylmethyl)-3-(4-(benzyloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d8):

[0067]

[0068] The intermediate compound used in the synthesis is 21. The product is a white solid powder with a yield of 94%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.97 (d, J =7.8 Hz, 2H), 7.45 (d, J =7.5 Hz, 2H), 7.38 (t, J =7.4 Hz,2H), 7.32 (t, J =7.3 Hz, 1H), 7.11 (d, J =7.8 Hz, 2H), 5.15 (s, 2H), 4.27 (t,J =9.7 Hz, 2H), 4.10–4.04 (m, 2H), 3.35 (d, J =7.6 Hz, 2H), 1.28 (s, 1H). 13 CNMR(200 MHz, Methanol- d 4 ) d 176.97, 167.85, 161.43, 136.81, 128.62, 128.20,127.69, 127.26, 114.98, 50.73, 28.54. IR (KBr, cm -1 ): 2934, 1697, 1616, 1591,1574, 1422, 1364, 1260, 1198, 1165, 1128, 1043. HRMS (ESI): m / z Calculate for C 19 H 19 N 3 O 2 [M+H] + : 322.1477. found: 322.1497.

[0069] Example 9: 4-(5-(azetidin-3-ylmethyl)-3-(4-((4-chlorobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d9):

[0070]

[0071] The intermediate compound used in the synthesis is 22. The product is a white solid powder with a yield of 92%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.98 (d, J =7.9 Hz, 2H), 7.45 (d, J =7.7 Hz, 2H), 7.39 (d, J =7.7 Hz,2H), 7.12 (d, J =7.9 Hz, 2H), 5.15 (s, 2H), 4.28 (t, J =9.7 Hz, 2H), 4.10–4.06(m, 2H), 3.37 (d, J =7.6 Hz, 2H), 1.28 (s, 1H).13 CNMR (200 MHz, Methanol- d 4 ) d 167.81, 161.22, 135.67, 128.83, 128.65, 128.31, 114.99, 68.88, 50.77, 28.54.IR (KBr, cm -1 ): 2918, 1674, 1592, 1422, 1378, 1263, 1203, 1177, 1138, 1092,864. HRMS (ESI): m / z calcd for C 19 H 18 C1N 3 O 2 [M+H] + : 356.1088. found: 356.1117.

[0072] Example 10: 4-(5-(azetidin-3-ylmethyl)-3-(4-((3,4-dichlorobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d10):

[0073]

[0074] The intermediate compound used in the synthesis is 23. The product is a white solid powder with a yield of 95%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.99 (d, J =7.8 Hz, 2H), 7.64 (s, 1H), 7.57–7.51 (m, 1H), 7.40 (d, J =8.2 Hz, 1H), 7.15–7.11 (m, 2H), 5.16 (s, 2H), 4.28 (t, J =9.7 Hz, 2H), 4.13–4.00 (m, 2H), 3.36 (d, J =7.6 Hz, 2H), 1.29 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d160.99, 137.77, 131.40, 130.37, 129.08, 128.70, 119.34, 115.00, 68.14,50.77, 29.41, 28.52. IR (KBr, cm -1 ): 3027, 1717, 1685, 1592, 1460, 1258,1203, 808, 721. HRMS (ESI): m / z calcd for C 19 H 17 Cl 2 N 3 O 2 [M+H] + : 390.0698. found:390.0718.

[0075] Example 11: 4-(5-(azetidin-3-ylmethyl)-3-(4-((3,4-difluorobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d11):

[0076]

[0077] The intermediate compound used in the synthesis is 24. The product is a white solid powder with a yield of 94%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.98 (d, J =8.1 Hz, 2H), 7.40–7.36 (m, 1H), 7.27 (t, J =6.8 Hz, 2H),7.12 (d, J =7.9 Hz, 2H), 5.13 (s, 2H), 4.29 (t, J =9.2 Hz, 2H), 4.08 (t, J =8.5 Hz,2H), 3.36 (d, J =7.2 Hz, 2H), 1.28 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 177.02,167.78, 161.02, 150.86, 134.47, 128.68, 123.69, 119.28, 117.02 (d, J=17.7 Hz),116.18 (d, J =18.3 Hz), 114.97, 68.34, 50.72, 29.35. IR (KBr, cm -1 ): 2934,1680, 1614, 1592, 1466, 1391, 1263, 1196, 1181, 1129, 1110. HRMS (ESI): m / zcalcd for C 19 H 17 F 2 N 3 O 2 [M+H] + :358.1289. found: 358.1305.

[0078] Example 12: 4-(5-(azetidin-3-ylmethyl)-3-(4-((3-fluorobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d12):

[0079]

[0080] The intermediate compound used in the synthesis is 25. White solid powder, yield 93%, 1 HNMR (800 MHz, Methanol- d 4 ) d 7.98 (d, J =7.7 Hz, 2H), 7.39 (q, J =7.0 Hz, 1H), 7.27 (d, J =7.6 Hz, 1H), 7.20(d, J =9.7 Hz, 1H), 7.12 (d, J =7.8 Hz, 2H), 7.05 (t, J =8.5 Hz, 1H), 5.17 (s, 2H), 4.29 (t, J =9.7 Hz, 2H), 4.11–4.05 (m, 2H), 3.36 (d, J =7.6 Hz, 2H), 3.31 (s,1H). 13 CNMR (200 MHz, Methanol- d 4 ) d176.98, 139.70, 130.00, 128.66, 122.76,119.19, 114.97, 114.32, 114.21, 113.76, 68.81, 50.70, 29.40, 28.52. IR ( KBr,cm -1 ): 2990, 2688, 1707, 1614, 1592, 1427, 1362, 1263, 1175, 1137, 836, 726.HRMS (ESI): m / z calcd for C 19 H 18 FN 3 O 2 [M+H] + : 340.1383. found: 340.1403.

[0081] Example 13: 4-(5-(azetidin-3-ylmethyl)-3-(4-((4-fluorobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d13):

[0082]

[0083] The intermediate compound used in the synthesis is 26. The product is a white solid powder with a yield of 95%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.97(d, J =7.8 Hz, 2H), 7.49–7.45(m, 2H), 7.13–7.08 (m, 4H), 5.13(s, 2H), 4.28 (t, J =9.7 Hz, 2H), 4.10–4.04(m, 2H), 3.36(d, J =7.6 Hz, 2H), 1.28(s, 1H). 13 CNMR(200 MHz, Methanol- d 4 ) d 167.82, 163.19, 161.98, 132.87, 129.42,129.38, 128.64, 119.07, 114.96, 114.94, 114.83, 69.01, 50.73, 29.45. IR (KBr, cm -1): 3219, 2693, 1680, 1614, 1592, 1362, 1263, 1203, 1175, 1132, 836,726. HRMS (ESI): m / z calcd for C 19 H 18 FN 3 O 2 [M+H] + : 340.1383. found: 340.1408.

[0084] Example 14: 4-(5-(azetidin-3-ylmethyl)-3-(4-((4-bromobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d14):

[0085]

[0086] The intermediate compound used in the synthesis is 27. The product is a white solid powder with a yield of 93%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.98 (d, J =7.9 Hz, 2H), 7.54 (d, J =7.6 Hz, 2H), 7.38 (d, J =7.7 Hz,2H), 7.11 (d, J =7.9 Hz, 2H), 5.13 (s, 2H), 4.28 (t, J =9.7 Hz, 2H), 4.10–4.06(m, 2H), 3.36 (d, J =7.6 Hz, 2H), 1.28 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 167.80, 161.19, 131.32, 129.10, 128.66, 114.98, 50.72, 29.40, 28.53. IR (KBr, cm -1 ): 2989, 2682, 1707, 1614, 1575, 1493, 1427, 1256, 1203, 1170, 1143.HRMS (ESI): m / z calcd for C 19 H18 Bn 3 O 2 [M+H] + : 400.0582. found: 400.0604.

[0087] Example 15: 4-(5-(azetidin-3-ylmethyl)-3-(4-((3,4,5-trifluorobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d15):

[0088]

[0089] The intermediate compound used in the synthesis is 28. The product is a white solid powder with a yield of 91%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.99 (d, J =8.1 Hz, 2H), 7.25 (t, J =7.3 Hz, 2H), 7.12 (d, J =8.1 Hz,2H), 5.14 (s, 2H), 4.29 (t, J =9.7 Hz, 2H), 4.11–4.06 (m, 2H), 3.36 (d, J =7.6Hz, 2H), 3.31 (s, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d 177.04, 167.75, 160.76,128.71, 119.50, 114.98, 111.25, 111.23, 111.16, 111.14, 29.40, 28.53. IR (KBr, cm -1 ): 3060, 1674, 1532, 1422, 1208, 1044, 852, 721. HRMS (ESI): m / zcalcd for C 19 H 16 F 3 N 3 O 2 [M+H] + : 376.1195. found: 376.1217.

[0090] Example 16: 4-(5-(azetidin-3-ylmethyl)-3-(4-((4-(trifluoromethyl)benzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d16):

[0091]

[0092] The intermediate compound used in the synthesis is 29. The product is a white solid powder with a yield of 92%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.99 (d, J =7.7 Hz, 2H), 7.70–7.63 (m, 4H), 7.14 (d, J =7.7 Hz, 2H),5.26 (s, 2H), 4.29 (t, J =9.7 Hz, 2H), 4.10–4.06 (m, 2H), 3.36 (d, J =7.6 Hz,2H), 3.31 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 177.01, 141.45, 128.70,127.45, 125.11, 125.09, 125.07, 125.05, 119.31, 68.75, 50.70, 28.52. IR (KBr, cm -1 ): 3016, 2682, 1702, 1427, 1329, 1148, 1071, 852, 726. HRMS (ESI): m / z calcd for C 20 H 18 F 3 N 3 O 2 [M+H] + : 390.1351. found: 390.1363.

[0093] Example 17: 4-(5-(azetidin-3-ylmethyl)-3-(4-((3,5-bis(trifluoromethyl)benzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d17):

[0094]

[0095] The intermediate compound used in the synthesis is 30. The product is a white solid powder with a yield of 93%. 1 HNMR (800 MHz, Methanol- d 4 ) d 8.08 (s, 2H), 8.02 (d, J =7.8 Hz, 2H), 7.94 (s, 1H), 7.19 (d, J =7.8Hz, 2H), 5.35 (s, 2H), 4.29 (t, J =9.7 Hz, 2H), 4.11–4.07 (m, 2H), 3.37 (d, J =7.6 Hz, 2H), 3.31 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 177.06, 167.74,160.73, 140.40, 131.63, 131.47, 128.77, 127.40, 122.74, 121.27, 119.65,114.98, 67.98, 50.70. IR ( KBr, cm -1 ):3005, 2677, 1680, 1614, 1493, 1367,1280, 1170, 1126, 847. HRMS (ESI): m / z calcd for C 21 H 17 F 6 N 3 O 2 [M+H] + :458.1225.found: 458.1236.

[0096] Example 18: 4-(5-(azetidin-3-ylmethyl)-3-(4-((3-(carbonitrile)benzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d18):

[0097]

[0098] The intermediate compound used in the synthesis is 31. The product is a white solid powder with a yield of 95%. 1 HNMR (800 MHz, Methanol- d 4 ) d7.99 (d, J =7.8 Hz, 2H), 7.83 (s, 1H), 7.78 (d, J =7.8 Hz, 1H), 7.69(d, J =7.7 Hz, 1H), 7.58 (t, J =7.7 Hz, 1H), 7.14 (d, J =7.9 Hz, 2H), 5.22 (s, 2H), 4.29 (t, J =9.7 Hz, 2H), 4.11–4.06 (m, 2H), 3.38–3.33 (m, 2H), 3.31 (s, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d 167.77, 160.93, 138.79, 131.72, 129.38, 128.72,119.40, 118.17, 114.98, 112.27, 68.38. IR (KBr, cm -1 ):2994, 2671, 2233, 1696,1597, 1214, 1022, 803. HRMS (ESI): m / z calcd for C 20 H 18 N 4 O 2 [M+H] + : 347.1430.found: 347.1451.

[0099] Example 19: 5-(azetidin-3-ylmethyl)-3-(4-(nonyloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d19):

[0100]

[0101] The intermediate compound used in the synthesis is 32. The product is a white solid powder with a yield of 92%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.95 (d, J =8.0 Hz, 2H), 7.02 (d, J =8.0 Hz, 2H), 4.28 (t, J=9.7 Hz,2H), 4.10–4.06 (m, 2H), 4.04 (t, J =6.3 Hz, 2H), 3.36 (d, J =7.6 Hz, 2H), 1.82–1.77 (m, 2H), 1.51–1.46 (m, 2H), 1.41–1.26 (m, 11H), 0.90 (t, J =6.8 Hz, 3H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.90, 167.90, 161.86, 114.51, 67.85, 50.72,29.41, 29.29, 29.11, 29.01, 28.90, 28.53, 25.73, 22.34. IR (KBr, cm -1 ): 2934,2852, 2699, 1712, 1592, 1258, 1143. HRMS (ESI): m / z calcd for C 21 H 31 N 3 O 2 [M+H] + :358.2416.found: 358.2433.

[0102] Example 20: 5-(azetidin-3-ylmethyl)-3-(4-(hexyloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d20):

[0103]

[0104] The intermediate compound used in the synthesis is 33. The product is a white solid powder with a yield of 93%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.95 (d, J =8.0 Hz, 2H), 7.02 (d, J =8.0 Hz, 2H), 4.29 (t, J =9.8 Hz,2H), 4.10–4.06 (m, 2H), 4.03 (t, J =6.4 Hz, 2H), 3.36 (d, J=7.6 Hz, 2H), 3.31(s, 1H), 1.82–1.76 (m, 2H), 1.52–1.46 (m, 2H), 1.42–1.34 (m, 4H), 0.93 (t, J =6.4 Hz, 3H). 13 CNMR (200 MHz, Methanol- d 4 ) d 161.85, 128.60, 118.52, 114.50,50.71, 31.36, 29.41, 28.87, 28.53, 25.43, 22.28, 12.97. IR (KBr, cm -1 ):2945,2682, 1707, 1570, 1433, 1170, 841. HRMS (ESI): m / z calcd for C 18 H 25 N 3 O 2 [M+H] + :316.1947. found: 316.1969.

[0105] Example 21: 5-(azetidin-3-ylmethyl)-3-(4-(ethoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d21):

[0106]

[0107] The intermediate compound used in the synthesis is 34. The product is a white solid powder with a yield of 91%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.95 (d, J =7.6 Hz, 2H), 7.02 (d, J =7.7 Hz, 2H), 4.28 (t, J =9.7 Hz,2H), 4.09 (dd, J =19.8, 8.9 Hz, 4H), 3.36 (d, J =7.6 Hz, 2H), 1.41 (t, J =6.9 Hz,3H), 1.28 (s, 1H). 13 CNMR (200 MHz, Methanol-d 4 ) d 176.92, 167.89, 161.69,118.54, 114.49, 63.37, 50.71, 29.41, 28.52, 13.63. IR (KBr, cm -1 ): 2995,2688, 1690, 1597, 1427, 1219, 1049, 846. HRMS (ESI): m / z calcd for C 14 H 17 N 3 O 2 [M+H] + : 260.1321. found: 260.1349.

[0108] Example 22: 5-(azetidin-3-ylmethyl)-3-(4-(cyclohexylmethoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d22):

[0109]

[0110] The intermediate compound used in the synthesis is 35. The product is a white solid powder with a yield of 93%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.95 (d, J =7.9 Hz, 2H), 7.02 (d, J =7.9 Hz, 2H), 4.28 (t, J =9.7 Hz,2H), 4.10–4.05 (m, 2H), 3.83 (d, J =6.3 Hz, 2H), 3.36 (d, J =7.6 Hz, 2H), 3.31(s, 1H), 1.88 (d, J =12.8 Hz, 2H), 1.79 (d, J =12.8 Hz, 3H), 1.72 (d, J =12.7 Hz,1H), 1.30 (dq, J =57.6, 12.7 Hz, 4H), 1.11 (q, J =11.1 Hz, 2H). 13 CNMR (200 MHz, Methanol-d 4 ) d 176.89, 167.91, 114.50, 73.27, 50.73, 37.66, 29.48, 29.41,25.55. IR (KBr, cm -1 ): 2940, 1669, 1592, 1422, 1362, 1263, 1175, 1137, 1027.HRMS (ESI): m / z calcd for C 19 H 25 N 3 O 2 [M+H] + : 328.1947. found: 328.1963.

[0111] Example 23: 5-(azetidin-3-ylmethyl)-3-(4-(cyclopentylmethoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d23):

[0112]

[0113] The intermediate compound used in the synthesis is 36. The product is a white solid powder with a yield of 90%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.95 (d, J =7.8 Hz, 2H), 7.02 (d, J =7.9 Hz, 2H), 4.28 (t, J =9.7 Hz,2H), 4.10–4.06 (m, 2H), 3.91 (d, J =6.9 Hz, 2H), 3.36 (d, J =7.6 Hz, 2H), 3.31(s, 1H), 2.38 (p, J =7.5 Hz, 1H), 1.86 (dq, J =12.4, 7.1 Hz, 2H), 1.72–1.58 (m,4H), 1.40 (dq, J =14.0, 7.2 Hz, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d176.90,167.90, 161.98, 128.59, 114.51, 72.04, 50.72, 38.98, 29.41, 29.00, 28.53,25.03. IR (KBr, cm -1 ):2962, 2688, 1707, 1570, 1427, 1263, 1170, 1143, 847.HRMS (ESI): m / z calcd for C 18 H 23 N 3 O 2 [M+H] + : 314.1790. found: 314.1796.

[0114] Example 24: 5-(azetidin-3-ylmethyl)-3-(4-(4,4,4-trifluorobutoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d24):

[0115]

[0116] The intermediate compound used in the synthesis is 37. The product is a white solid powder with a yield of 90%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.97 (d, J =7.7 Hz, 2H), 7.06 (d, J =7.7 Hz, 2H), 4.29 (t, J =9.7 Hz,2H), 4.14–4.06 (m, 4H), 3.36 (d, J =7.6 Hz, 2H), 3.31 (s, 1H), 2.38 (dq, J =20.8,10.7 Hz, 2H), 2.05 (dt, J =14.8, 6.4 Hz, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.98, 167.83, 161.37, 128.66, 114.54, 50.70, 30.14, 29.99, 29.85, 29.71,29.40, 28.53, 21.81, 21.80. IR ( KBr, cm-1 ):2989, 2677, 1685, 1592, 1433,1263, 1203, 1132, 1022. HRMS (ESI): m / z calcd for C 16 H 18 F 3 N 3 O 2 [M+H] + : 342.1351.found: 342.1352.

[0117] Example 25: 5-(azetidin-3-ylmethyl)-3-(4-(butoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d25):

[0118]

[0119] The intermediate compound used in the synthesis is 38. The product is a white solid powder with a yield of 89%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.95 (d, J =7.7 Hz, 2H), 7.02 (d, J =7.7 Hz, 2H), 4.28 (t, J =9.7 Hz,2H), 4.10–4.02 (m, 4H), 3.36 (d, J =7.6 Hz, 2H), 3.31 (s, 2H), 1.78 (p, J =6.7Hz, 2H), 1.52 (h, J =7.4 Hz, 2H), 1.00 (t, J =7.4 Hz, 3H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.91, 167.90, 161.86, 114.49, 67.55, 50.71, 30.99, 29.41,28.52, 18.87. IR (KBr, cm -1 ): 2962, 2682, 1707, 1575, 1427, 1269, 1143, 836,726. HRMS (ESI): m / z calcd for C16 H 21 N 3 O 2 [M+H] + : 288.1634. found: 288.1644.

[0120] Example 26: 4-(5-(azetidin-3-ylmethyl)-3-(4-((4-(carbonitrile)benzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d26):

[0121]

[0122] The intermediate compound used in the synthesis is 39. The product is a white solid powder with a yield of 93%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.99 (d, J =7.9 Hz, 2H), 7.75 (d, J =7.6 Hz, 2H), 7.65 (d, J =7.8 Hz,2H), 7.13 (d, J =8.0 Hz, 2H), 5.26 (s, 2H), 4.28 (t, J =9.7 Hz, 2H), 4.10–4.04(m, 2H), 3.36 (d, J =7.6 Hz, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d 132.12, 128.72,127.66, 68.62, 50.73, 47.93, 47.82, 29.40, 28.54. IR (KBr, cm -1 ):2978, 2227,1680, 1592, 1460, 1427, 1197, 1175, 1132, 819, 721. HRMS (ESI): m / z calcd forC 20 H 18 N 4 O 2 [M+H] + : 347.1430. found: 347.1436.

[0123] Example 27: 5-(azetidin-3-ylmethyl)-3-(4-(3-bromopropoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d27):

[0124]

[0125] The intermediate compound used in the synthesis is 40. The product is a white solid powder with a yield of 94%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.97 (t, J =8.3 Hz, 2H), 7.06 (t, J =7.6 Hz, 2H), 4.29 (t, J =9.8 Hz,2H), 4.21–4.14 (m, 2H), 4.11–4.06 (m, 2H), 3.64 (t, J =6.4 Hz, 2H), 3.36 (d, J =7.6 Hz, 2H), 3.31 (s, 2H), 2.32 (p, J =6.0 Hz, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.96, 167.84, 161.46, 128.66, 114.57, 65.40, 50.87, 50.71, 32.06, 29.40,29.05, 28.53. IR (KBr, cm -1 ): 3022, 1707, 1614, 1433, 1356, 1208, 1137, 847,721. HRMS (ESI): m / z calcd for C 15 H 18 Bn 3 O 2 [M+H] + : 352.0582. found: 352.0609.

[0126] Example 28: 5-(azetidin-3-ylmethyl)-3-(4-(3-(benzyloxy)propoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d28):

[0127]

[0128] The intermediate compound used in the synthesis is 41. The product is a white solid powder with a yield of 86%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.96 (d, J =8.1 Hz, 2H), 7.33–7.28 (m, 4H), 7.25 (t, J =7.0 Hz, 1H),7.02 (d, J =8.1 Hz, 2H), 4.52 (s, 2H), 4.28 (t, J =9.8 Hz, 2H), 4.15 (t, J =6.2 Hz,2H), 4.12–4.06 (m, 2H), 3.67 (t, J =6.1 Hz, 2H), 3.36 (d, J =7.6 Hz, 2H), 3.31(s, 2H), 2.08 (p, J =6.1 Hz, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d 161.71, 138.36,128.61, 127.99, 127.43, 127.27, 118.67, 114.56, 66.25, 64.73, 50.72, 29.41,29.25. IR (KBr, cm -1 ): 2962, 2677, 1680, 1614, 1427, 1263, 1208, 1181, 1126,836, 726. HRMS (ESI): m / z calcd for C 22 H 25 N 3 O 3 [M+H] + : 380.1896. found:380.1913.

[0129] Example 29: 5-(azetidin-3-ylmethyl)-3-(4-(3-phenylpropoxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d29):

[0130]

[0131] The intermediate compound used in the synthesis is 42. The product is a white solid powder with a yield of 95%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.96 (d, J =8.2 Hz, 2H), 7.26 (t, J =7.3 Hz, 2H), 7.21 (d, J =7.5 Hz,2H), 7.16 (t, J =7.2 Hz, 1H), 7.01 (d, J =8.2 Hz, 2H), 4.28 (t, J =9.7 Hz, 2H),4.12–4.06 (m, 2H), 4.02 (t, J =6.1 Hz, 2H), 3.36 (d, J =7.6 Hz, 2H), 3.31 (s,1H), 2.81 (t, J =7.6 Hz, 2H), 2.09 (p, J =6.7 Hz, 2H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.92, 161.74, 141.41, 128.62, 128.16, 128.07, 125.60, 118.64, 114.53,66.80, 30.68, 29.41, 28.53. IR (KBr, cm -1 ): 2962, 2693, 1707, 1592, 1367,1256, 1175, 1143, 841, 726. HRMS (ESI): m / z calcd for C 21 H 23 N 3 O 2 [M+H] + :350.1790. found: 350.1807.

[0132] Example 30: 5-(azetidin-3-ylmethyl)-3-(4-(8-en-1-yloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d30):

[0133]

[0134] The intermediate compound used in the synthesis is 43. The product is a white solid powder with a yield of 93%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.95 (d, J =7.7 Hz, 2H), 7.02 (d, J =7.8 Hz, 2H), 5.81 (td, J =16.9,6.8 Hz, 1H), 4.98 (d, J =17.1 Hz, 1H), 4.91 (d, J =10.2 Hz, 1H), 4.28 (t, J =9.7Hz, 2H), 4.11–4.06 (m, 2H), 4.04 (t, J =6.3 Hz, 2H), 3.36 (d, J =7.6 Hz, 2H),3.31 (s, 2H), 2.05 (q, J =7.0 Hz, 2H), 1.79 (p, J =6.7 Hz, 2H), 1.49 (p, J =7.4 Hz,2H), 1.39 (ddd, J =29.6, 14.4, 6.9 Hz, 7H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.90,161.85, 128.60, 118.52, 114.51, 113.37, 29.41, 28.92, 28.87, 28.74, 28.64,28.53, 25.69. IR (KBr, cm -1 ): 2995, 2934, 2697, 1712, 1597, 1422, 1367, 1263,1175, 1137, 726. HRMS (ESI): m / z calcd for C 21 H 29 N 3 O 2 [M+H] +: 356.2260. found:356.2274.

[0135] Example 31: 5-(azetidin-3-ylmethyl)-3-(4-(9-en-1-yloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d31):

[0136]

[0137] The intermediate compound used in the synthesis is 44. The product is a white solid powder with a yield of 95%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.96 (d, J =7.8 Hz, 2H), 7.02 (d, J =7.8 Hz, 2H), 5.80 (td, J =16.8,6.8 Hz, 1H), 4.98 (d, J =17.1 Hz, 1H), 4.28 (t, J =9.8 Hz, 2H), 4.10–4.02 (m,4H), 3.36 (d, J =7.6 Hz, 2H), 3.31 (s, 3H), 2.05 (q, J =7.1 Hz, 2H), 1.80 (p, J =6.5 Hz, 2H), 1.49 (p, J =7.3 Hz, 2H), 1.42–1.27 (m, 9H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.89, 161.86, 128.60, 114.51, 67.84, 50.73, 33.49, 29.04,28.88, 28.75, 28.70, 28.53. IR (KBr, cm -1 ): 2934, 2856, 1701, 1592, 1477,1422, 1367, 1258, 1192, 1143, 726. HRMS (ESI): m / z calcd for C 22 H 31 N 3 O 2 [M+H]+ :370.2416. found: 370.2432.

[0138] Example 32: 4-(5-(azetidin-3-ylmethyl)-3-(4-((3-chlorobenzyl)oxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d32):

[0139]

[0140] The intermediate compound used in the synthesis is 45. The product is a white solid powder with a yield of 91%. 1 HNMR (800 MHz, Methanol- d 4 ) d 7.98 (d, J =7.7 Hz, 2H), 7.48 (s, 1H), 7.37 (q, J =8.0, 7.5 Hz, 2H),7.33 (d, J =7.3 Hz, 1H), 7.12 (d, J =7.8 Hz, 2H), 5.16 (s, 2H), 4.28 (t, J =9.7 Hz,2H), 4.12–4.06 (m, 2H), 3.36 (d, J =7.5 Hz, 2H), 3.31 (s, 1H). 13 CNMR (200 MHz, Methanol- d 4 ) d 176.98, 167.80, 139.24, 134.10, 129.78, 128.67, 127.68, 127.03,119.22, 114.97, 68.77, 50.71, 29.40, 28.52. IR ( KBr, cm -1 ): 3027, 2671, 1674,1592, 1427, 1362, 1186, 1148, 841, 803, 721. HRMS (ESI): m / z calcd forC 19 H 18 C1N 3 O 2 [M+H] + : 356.1088. found: 356.1108.

[0141] Example 33: 5-(azetidin-3-ylmethyl)-3-(4-(octyloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (d33):

[0142]

[0143] The intermediate compound used in the synthesis is 46. The product is a white solid powder with a yield of 95%, mp: 135.6 ℃-136.2 ℃, 1 HNMR (600 MHz, DMSO- d 6 ) δ(ppm): 9.04 (s, 1H), 8.88 (s,1H), 7.93–7.89(m, 2H), 7.11–7.06 (m, 2H), 4.16–4.08 (m, 2H), 4.03 (t, J =6.5 Hz, 2H), 3.95–3.87 (m, 2H), 3.39–3.32 (m, 3H), 1.77–1.69 (m, 2H), 1.47–1.36 (m, 2H), 1.35–1.22 (m, 8H), 0.90–0.83 (m, 3H). 13 CNMR (100 MHz, DMSO) δ(ppm): 177.93, 167.65,161.64, 158.99 (q, J =33.0 Hz, 1C), 129.15 (2C), 118.67, 115.50 (2C), 68.19,50.58 (2C), 31.69, 29.45, 29.19, 29.14, 29.12, 29.02, 25.93, 22.54, 14.38.HRMS (ESI): m / z calcd for C 20 H 30 N 3 O 2 [M+H] + : 344.2338. found: 344.2331.

[0144] Experimental Example 1: Inhibition rate and IC of the compounds of the present invention on SphK 1 and SphK 2 50 Activity experiment

[0145] 1. Experimental methods

[0146] Dissolve the compound in pure DMSO to prepare a 10 mM stock solution and use kinase buffer (pH = 7.4, composition: 40 mM / LTris, 10 mM / LMgCl 2 , 0.1g / LBSA, 1mM / LDTT, 10µM / LATP). SphK 1 / 2 was added to a 96-well plate and then the desired concentration of compound (single concentration: 10µM, IC 50 The test concentrations were: 0.01, 0.1, 1, 10 and 100 µM) for 40 min at 30 °C. Then the ATP test solution was added and the mixture was incubated at room temperature for 5 min. Luminescence was immediately measured using a microplate spectrophotometer (AD340, Beckman, USA). Data were analyzed using Graphpad Prism 7.022 to calculate the single concentration inhibition rate or IC 50 value.

[0147] 2. The experimental results are shown in the table below.

[0148] Inhibition rate of compounds on SphKs

[0149]

[0150] The tested compounds showed certain inhibitory activity at 10 μM, among which most compounds had an inhibition rate of more than 20% on SphK2. Therefore, some compounds were selected to determine the IC 50 .

[0151] 3. IC of some compounds against SphK2 50 result

[0152] IC of compounds against SphK2 50

[0153]

[0154] The IC values ​​of all tested compounds, except d9, for SphK2 50 All of them were less than 10 μM, which is valuable for anti-tumor effect research.

[0155] Experimental Example 2: Experimental study on the inhibitory activity of the compounds of the present invention on Hela, DU145, HT29, and SW620 tumor cells

[0156] 1. Experimental Materials

[0157] The compounds d19, d30, d31, and d33 were dissolved in dimethyl sulfoxide (DMSO, final concentration 0.4%) and prepared to 1 mg / mL in RPMI-1640 medium containing 15% fetal bovine serum for later use. When grouping and dosing were performed, the compounds were diluted in multiples with the medium to the required concentration.

[0158] Cell lines: cervical cancer (Hela), prostate cancer (DU145), colon cancer (HT-29), colon cancer (SW620), all purchased from the Chinese Academy of Sciences Cell Bank, using DMEM (High Glucose) culture medium containing 15% fetal bovine serum, placed at 37°C, 5% CO 2 Culture in an incubator.

[0159] 2. Experimental methods:

[0160] Hela cells, DU145 cells, HT-29 cells, SW620 cells (6×10 3 After 24 h of incubation, test compounds (0.1, 0.3, 1, 3, 10, 30, and 100 μM) and cisplatin (0.1, 0.3, 1, 3, 10, 30, and 100 μM) were added to each well of a 96-well culture plate. The culture supernatant was removed and 100 μL of basal medium containing 0.5 mg / mL CCK-8 was added to each well. The plates were incubated at 37°C and 5% CO. 2 The incubation was continued for 2.5 h under the same conditions, and the absorbance (OD) was measured at 450 nm using a multifunctional microplate reader. Cell activity (100%) = OD value of the drug group / OD value of the blank group*100%.

[0161] 3. Statistical analysis:

[0162] GraphPad Prism 9 software was used for statistical drawing. Data were expressed as mean ± standard deviation (Mean ± SD). Differences between groups were analyzed by one-way ANOVA followed by Dunnett's test. P <0.05 indicated a statistically significant difference.

[0163] 4. Experimental results:

[0164] Effects of four compounds on the survival rate of Hela cells

[0165]

[0166] Effects of four compounds on the survival rate of DU145 cells

[0167]

[0168] Effects of three compounds on the survival rate of HT-29 cells

[0169]

[0170] Effects of three compounds on the survival rate of SW620 cells

[0171]

[0172] By observing the above experimental data, it is found that the designed and synthesized series of compounds have ideal effects on SphK2 proliferation and cell proliferation inhibition, and are candidate compounds for clinical research.

Claims

1. A compound represented by the general formula (I), Formula (I) in, R is , , , , , , , , 、 、 、 、 、 、 、 、 、 、 。 2. The compound according to claim 1, characterized in that The R is , , , , , , , , , , .

3. The compound according to claim 2, characterized in that The R is , , , .

4. The compound according to claim 3, characterized in that The R is , , .

5. A method for preparing the compound according to any one of claims 1 to 4, comprising the following steps: The intermediate compound 14-46 and trifluoroacetic acid were dissolved in dichloromethane, and the mixture was stirred at room temperature for 1 hour and then detected by TLC. After the reaction was completed, the reaction solution was dried by rotary evaporation and the crude product was separated by silica gel column chromatography to obtain d series substituted oxadiazole compounds with a yield of 85-95%.

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

7. Use of the compound according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 6 in the preparation of a medicament for preventing and / or treating a disease mediated by SphK2.

8. The use according to claim 7, characterized in that: The SphK2-mediated diseases are colon cancer, cervical cancer and prostate cancer.

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