1, 2, 4-oxadiazole compound containing nitrogen heterocyclic ring and application thereof

Abstract

The invention relates to the field of medicinal chemistry, in particular to 1, 2, 4-oxadiazole compounds containing nitrogen heterocyclic rings and application of the 1, 2, 4-oxadiazole compounds. Pharmacological tests show that the compound disclosed by the invention has a relatively strong inhibition effect on both SphK1 and SphK2, and can act on the SphK1 and the SphK2 at the same time. An activity result shows that in the inhibitory activity on A549 and MDA-MB-231 cells, the activity of all the compounds is superior to that of cis-platinum; in the inhibitory activity to HCT116 cells, the activity of T16, T18, T30, T32, T33, T42, T47 (R), T48 (R), T55 (S) and T57 (S) is superior to that of cis-platinum.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and particularly relates to a class of 1,2,4-oxadiazole compounds containing nitrogen heterocycles, their pharmaceutical compositions and their applications. Background Art

[0002] Cancer is a complex disease caused by abnormal cell proliferation and is also one of the major global public health challenges. Normally, human cells maintain tissue function balance through orderly cell division and apoptosis. However, when cancer occurs, gene mutations cause uncontrolled cell division, forming abnormal tissues (tumors). These tumors are divided into benign (non-spreading) and malignant (invading surrounding tissues and metastasizing through the blood or lymphatic system). According to the World Health Organization, lung cancer, breast cancer, colorectal cancer, etc. are the most common current cancer types. Cancer is one of the most difficult diseases to overcome in the world today. Since the number of people dying from cancer is increasing year by year, the research on the occurrence, development, and metastasis mechanisms of cancer and the development of cancer treatment drugs have always been hot topics in domestic and international research. The 5-year survival rate of Chinese cancer patients has been significantly improved, and this progress benefits from the wide promotion of tumor screening and multidisciplinary comprehensive treatment, the implementation of precision oncology, and the continuous innovation of new drug research and development.

[0003] Sphingolipids are ubiquitous components in the biological membranes of eukaryotic cells. Sphingolipids and their metabolites are involved in many important signal transduction pathways. Among them, sphingosine kinases (SphKs) can catalyze the production of sphingosine 1-phosphate (S1P) from sphingosine. Sphingosine-1-phosphate (S1P) regulates a variety of biological processes, including cell proliferation and differentiation and immune cell trafficking. There are two subtypes of SphKs, SphK1 and SphK2, which have high homology in molecular structure, different tissue distributions and functions. Inhibiting one subtype can play a role in treating related diseases, and inhibiting both subtypes may achieve a wider treatment range and better drug efficacy. Currently, the mechanism of SphK1 has been studied more deeply, but the physiological function of SphK2 is more complex, and its role in pathological processes has only gradually been reported in recent years. The expression and activity of SphK1 and SphK2 in a variety of tumor cells are significantly higher than those in normal tissues or cells, and can inhibit apoptosis, promote cell proliferation and angiogenesis by increasing the synthesis of S1P. The SphKs / S1P signaling axis plays an important role in the regulation of tumor cell proliferation and apoptosis. Inhibiting the expression and activity of SphKs can regulate the biological behavior of tumors and the response to drug treatment, and improve the prognosis of patients. Some current SphKs inhibitors, such as the SphK2 inhibitor ABC294640, have ideal oral bioavailability and pharmacokinetic properties and have now entered clinical phase II research, indicating that the application of SphKs inhibitors in cancer treatment is gradually moving from the laboratory to the clinic.

[0004] Patent CN119119016A discloses a series of substituted oxadiazole compounds, their preparation methods and uses. Pharmacological tests show that this series of compounds has strong selectivity and inhibitory effects on SphK2, and has anti-proliferative effects on cervical cancer cells, prostate cancer cells and colon cancer cells. The compounds obtained in this patent are SphK2 selective inhibitors. If compounds that can inhibit both SphK1 and SphK2 can be obtained, their scope of application may be wider and the value of drug development may be better. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a class of 1,2,4-oxadiazole compounds containing nitrogen heterocycles, their preparation methods, pharmaceutical compositions and their applications in the preparation of preventing and / or treating diseases mediated by SphKs.

[0006] To solve the technical problems of the present invention, the present invention provides the following technical solutions: The first aspect of the technical solution of the present invention is to provide a compound represented by the general formula (I), its solvate: Formula (I) Wherein, n is 1 or 2, R 1 is Cl, F, Br, CH3, CF3, R 2 is: .

[0007] Preferably, the 1,2,4-oxadiazole compound containing nitrogen heterocycle represented by the general formula (I) or its solvate: Formula (I) Wherein, n = 1, 2, R 1 = F, Br, CH3, CF3, R 2 is .

[0008] Preferably, the above-mentioned 1,2,4-oxadiazole compound containing nitrogen heterocycle or its solvate has the following structural formula: .

[0009] More preferably, the above-mentioned 1,2,4-oxadiazole compound containing nitrogen heterocycle or its solvate has the following structural formula: .

[0010] The term "solvate" refers to a complex formed by the coordination of the compound of the present invention with solvent molecules in a specific ratio.

[0011] The second aspect of the technical solution of the present invention is to provide a preparation method of the compound described in the first aspect. The compound of general formula (I) of the present invention can be prepared by the following method: 。

[0012] The substituted p-hydroxybenzonitrile reacts with hydroxylamine hydrochloride to obtain an intermediate aminooxime (2), which then undergoes a cyclization reaction with 1-Boc-azetidine-3-acetic acid / N-Boc-3-pyrrolidineacetic acid to obtain an oxadiazole intermediate (3). The obtained product is then treated with trifluoroacetic acid to obtain the target compound through Williamson synthesis with a halogenated hydrocarbon to obtain an ether intermediate (4).

[0013] The third aspect of the technical solution of the present invention is to provide a pharmaceutical composition comprising a 1,2,4-oxadiazole compound containing a nitrogen heterocycle shown in the first aspect or a solvate thereof and a pharmaceutically acceptable carrier, which is any pharmaceutically acceptable dosage form, preferably an oral preparation or an injection. It contains a physiologically effective amount of the compound shown in general formula (I) of 0.01 g to 10 g, which can be 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.04 g, 0.05 g, 0.1 g, 0.125 g, 0.2 g, 0.25 g, 0.3 g, 0.4 g, 0.5 g, 0.6 g, 0.75 g, 1 g, 1.25 g, 1.5 g, 1.75 g, 2 g, 2.5 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, 10 g, etc.

[0014] Any compound of the present invention can be administered to a patient in need of such treatment by oral or parenteral administration or other means.

[0015] The compounds described in the present invention can be made into common pharmaceutical preparations by adding pharmaceutically acceptable carriers, such as tablets, capsules, powders, syrups, liquids, suspensions, injections, and common pharmaceutical excipients such as flavors, sweeteners, liquid or solid fillers or diluents can be added.

[0016] The clinical dosage of the compounds of the present invention is 0.01 - 1000 mg / day, and this range can also be deviated from according to the severity of the condition or the difference in dosage form.

[0017] The fourth aspect of the technical solution of the present invention is to provide the use of a 1,2,4-oxadiazole compound containing a nitrogen heterocycle shown in the first aspect or a solvate thereof and the pharmaceutical composition described in the third aspect in the preparation of a drug for preventing and / or treating diseases mediated by SphK1 and / or SphK2.

[0018] The diseases mediated by SphK1 are cancer and inflammatory diseases. The cancers include colon cancer, lung cancer, breast cancer, liver cancer, gastric cancer, lung adenocarcinoma, melanoma, and esophageal cancer; the inflammatory diseases include inflammatory bowel disease, hepatitis, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, or multiple sclerosis.

[0019] The diseases mediated by SphK2 are cancer, inflammatory diseases, ischemia-reperfusion injury, renal fibrosis, Alzheimer's disease, diabetic retinopathy, and diabetic nephropathy. The cancers are rectal cancer, gastric cancer, renal cancer, colon cancer, uterine cancer, ovarian cancer, breast cancer, and lung cancer.

[0020] Preferably, the cancers are lung cancer, colon cancer, and breast cancer.

[0021] Beneficial technical effects: (1) The compounds of the present invention have strong inhibitory effects on both SphK1 and SphK2, can act on SphK1 and SphK2 simultaneously. Compared with selective inhibitors of SphK1 or SphK2, the compounds of the present invention may have a wider range of possible indications and higher drug development value.

[0022] (2) When tested at 10 μM, most of the 17 compounds had inhibition rates of more than 40% or even reached above 96% for SphK1 and SphK2. Among them, T16, T18, T23, T29, T30, T31, T32, T33, T35, T42, T44, T47(R), T48(R), T55(S), and T57(S) have strong inhibitory effects on phK1 and SphK2.

[0023] (3) The activity results showed that in the inhibitory activity against A549 cells, the compounds all showed better inhibitory effects than cisplatin. Especially, T16, T33, T42, T47(R), and T55(S) had the best activities, and the IC 50 were 4.12 ± 1.41 μM, 4.78 ± 0.18 μM, 4.60 ± 0.61 μM, 3.27 ± 0.25 μM, and 4.56 ± 0.45 μM respectively. In the inhibitory activity against HCT116 cells, the activities of T16, T18, T30, T32, T33, T42, T47(R), T48(R), T55(S), and T57(S) were all better than cisplatin, and the IC 50They were 5.37±0.62 μM, 5.27±0.22 μM, 4.32±0.11 μM, 4.29±0.11 μM, 4.13±0.21 μM, 4.14±0.26 μM, 3.87±0.06 μM, 5.43±0.23 μM, 4.11±0.21 μM, 3.74±0.14 μM respectively. In the inhibitory activity against MDA-MB-231 cells, the activities of all compounds were better than those of cisplatin, and the activities of T16 and T47(R) were particularly significant, with IC 50 They were 4.47±0.14 μM and 4.00±0.07 μM respectively. Detailed implementation manners

[0024] The present invention will be further described below in conjunction with the detailed implementation manners, so that those skilled in the art can better understand the present invention, but the present invention is not limited thereto.

[0025] Dissolve substituted p-hydroxybenzonitrile (11 mmol, 1.0 eq), hydroxylamine hydrochloride (16 mmol, 1.5 eq), and sodium bicarbonate (16 mmol, 1.5 eq) in methanol (20 mL). Reflux for 22 hours. Cool the reaction solution to room temperature and filter by suction. Evaporate the solvent to obtain a crude product of aminooxime. Dissolve the obtained crude product, 1-Boc-azetidine-3-acetic acid (18 mmol, 1.2 eq), and DIPEA (27 mmol, 1.8 eq) in DMF (20 mL), and add HATU (16 mmol, 1.1 eq). React at 100 o React at 12 hours. Extract the reaction solution with ethyl acetate. Wash the organic phase successively with water and saturated brine, dry over anhydrous sodium sulfate, filter, evaporate the solvent, and purify the crude product by silica gel column chromatography (mobile phase: n-hexane - ethyl acetate = 4:1) to obtain intermediate (3). Dissolve intermediate (3) (0.52 mmol, 1.0 eq), cesium carbonate (0.58 mmol, 1.1 eq), and halogenated hydrocarbon (0.58 mmol, 1.1 eq) in DMF (20 mL). React at 75 oReact for 8 hours. The reaction solution is extracted with ethyl acetate. The organic phase is washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, the solvent is evaporated, and the crude product is separated and purified by silica gel column chromatography (mobile phase: n-hexane - ethyl acetate = 10:1) to obtain intermediate (4). Dissolve intermediate compound (4) (3 mmol, 1.0 eq) in CH2Cl2 (6 mL), and slowly add TFA (3 mL) dropwise with stirring. React at room temperature for 4 hours. Evaporate the solvent under reduced pressure from the reaction solution, and the crude product is separated and purified by silica gel column chromatography (methylene chloride - methanol = 15:1) to obtain white target compounds T1 - T64 with a yield of 85% - 90% Example 1: 5-(azetidin-3-ylmethyl)-3-(3-fluoro-4-(nonyloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (T16): Dissolve substituted p-hydroxybenzonitrile 3-fluoro-4-hydroxybenzonitrile, hydroxylamine hydrochloride, and sodium bicarbonate in methanol and reflux for 22 hours. Cool the reaction solution to room temperature and filter by suction. Evaporate the solvent under reduced pressure to obtain a crude product of aminooxime. Dissolve the obtained crude product, 1-Boc-azetidine-3-acetic acid, and DIPEA in DMF, add HATU, 100 o React at 12 hours. The reaction solution is extracted with ethyl acetate, and the crude product is separated and purified by silica gel column chromatography. Then dissolve this intermediate, cesium carbonate, and halogenated hydrocarbon 1-bromononane in DMF. 75 o React at 8 hours. The reaction solution is extracted with ethyl acetate. The organic phase is washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, the solvent is evaporated under reduced pressure, and the crude product is separated and purified by silica gel column chromatography to obtain the key intermediate. Dissolve the key intermediate in CH2Cl2, and slowly add TFA dropwise with stirring. React at room temperature for 4 hours. After the reaction is completed, evaporate the solvent under reduced pressure from the reaction solution, and the crude product is separated and purified by silica gel column chromatography (methylene chloride - methanol = 15:1) to obtain white target compound T16 with a yield of 89%. 1 H NMR (800 MHz, DMSO- d6) δ 7.79 (d, J = 8.1 Hz, 1H), 7.75 (d, J = 11.6 Hz,1H), 7.34 (t, J = 8.7 Hz, 1H), 4.13 (d, J = 6.9 Hz, 4H), 3.93 (s, 2H), 3.39(d, J = 10.9 Hz, 3H), 1.76 (t, J = 7.2 Hz, 2H), 1.42 (s, 2H), 1.33 (s, 2H),1.30 – 1.24 (m, 8H), 0.85 (dd, J = 9.4, 4.6 Hz, 3H). 13 C NMR (200 MHz, DMSO- d 6)δ 178.32, 166.97, 158.90 (d, J = 34.3 Hz, 1C), 151.99 (d, J = 245.7 Hz, 1C),149.70 (d, J = 10.3 Hz, 1C), 124.59, 118.95 (d, J = 7.3 Hz, 1C), 117.65 (d, J= 299.3 Hz, 1C), 115.64, 114.81 (d, J = 19.8 Hz, 1C), 69.30(2C), 50.53, 31.73(2C), 29.39(2C), 29.16, 29.10, 28.88, 25.81, 22.56, 14.38. IR (KBr, cm -1 ):723.01, 791.47, 831.86, 1141.37, 1202.46, 1286.09, 1369.44, 1506.40, 1593.74,1698.83, 2687.61, 2857.02, 2931.20, 3433.71. HRMS (ESI): m / z calcd forC 21 H 30 FN3O2[M+H] + : 376.2400 found: 376.2360. The preparation method of the compounds in Example 2-10 is the same as that in Example 1, except that different substituted 4-hydroxybenzonitriles and halogenated hydrocarbons are used. The substituted 4-hydroxybenzonitriles and halogenated hydrocarbons used in each specific example are indicated in the corresponding example: Example 2: 5-(azetidin-3-ylmethyl)-3-(4-(dodec-9-en-1-yloxy)-3-fluorophenyl)-1,2,4-oxadiazole trifluoroacetate (T18): The substituted 4-hydroxybenzonitrile used was 3-fluoro-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromodecene. T18 is a white solid powder with a yield of 87%. 1 H NMR (800 MHz, DMSO- d 6) δ 7.78 (d, J = 8.2 Hz, 1H), 7.76– 7.72 (m, 1H), 7.34 (q, J = 8.7, 7.3 Hz, 1H), 5.79 (qt, J = 10.5, 5.8 Hz,1H), 4.99 (dd, J = 17.3, 2.1 Hz, 1H), 4.93 (d, J = 9.0 Hz, 1H), 4.12 (d, J =6.2 Hz, 4H), 3.92 (t, J = 7.8 Hz, 2H), 3.38 (d, J = 5.1 Hz, 3H), 2.01 (t, J =6.5 Hz, 2H), 1.75 (s, 2H), 1.42 (s, 2H), 1.37 – 1.26 (m, 8H). 13 C NMR (200 MHz,DMSO- d 6) δ 178.32, 166.97, 152.60, 151.38, 149.70 (d, J = 10.2 Hz, 1C),139.29, 124.59, 118.95 (d, J = 7.3 Hz, 1C), 117.69 (d, J = 294.8 Hz, 1C),115.68, 115.11, 114.82 (d, J = 20.3 Hz, 1C), 69.30, 50.54(2C), 33.64(2C),29.38, 29.24, 29.11, 28.90, 28.88, 28.70, 25.78. IR (KBr, cm -1): 720.57, 793.70, 832.19, 880.94, 911.93, 993.75, 1019.03, 1138.65, 1165.42, 1203.94, 1282.30, 1371.58, 1423.24, 1476.23, 1505.86, 1548.15, 1578.43, 1702.72, 2518.65, 2686.60, 2852.21, 2925.86. HRMS (ESI): m / z calcd for C 22 H 30 FN3O2[M + H] + : 388.2400 found: 388.2349. Example 3: 5-(Azetidin-3-ylmethyl)-3-(3-bromo-4-(oct-8-en-1-yloxy)benzene)-1,2,4-oxadiazole trifluoroacetate (T23): The substituted 4-hydroxybenzonitrile used was 3-bromo-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromoheptene. T23 is a white solid powder with a yield of 86%. 1 H NMR (800 MHz, Methanol- d 4) δ 8.18 (s, 1H), 7.98 (s, 1H), 7.14 (t, J = 9.1 Hz, 1H), 5.81 (td, J = 17.0, 7.5 Hz, 1H), 4.98 (d, J = 15.0 Hz, 1H), 4.91 (d, J = 10.2 Hz, 1H), 4.29 (t, J = 10.2 Hz, 2H), 4.14 – 4.06 (m, 4H), 3.54 (q, J = 8.8 Hz, 1H), 3.37 (d, J = 7.7 Hz, 2H), 2.05 (qt, J = 6.9, 2.7 Hz, 2H), 1.84 (ddd, J = 12.9, 10.5, 5.3, 3.3 Hz, 2H), 1.55 (tddd, J = 9.9, 7.7, 4.6, 2.1 Hz, 2H), 1.45 – 1.34 (m, 6H).13 C NMR (200 MHz, Methanol- d 4) δ 177.25, 166.86, 161.75 (d, J J = 36.9 Hz, 1C), 157.91, 138.71, 131.66, 127.69, 119.81, 116.68 (d, J J = 293.2 Hz, 1C), 113.34, 112.88, 111.85, 69.03, 50.69(2C), 33.46, 29.35, 28.79, 28.72, 28.66, 28.60, 28.52, 25.65. IR (KBr, cm -1 ): 724.26, 800.20, 840.45, 905.66, 992.97, 1048.13, 1138.12, 1209.25, 1273.11, 1295.33, 1328.94, 1390.97, 1429.47, 1464.71, 1559.14, 1595.36, 1686.17, 1716.86, 2852.80, 2929.13. HRMS (ESI): m / z calcd for C 21 H 28 BrN3O2[M+H] + : 434.1443 found: 434.1403. Example 4: 5-(azetidin-3-ylmethyl)-3-(4-(dodecyloxy)-3-methylphenyl)-1,2,4-oxadiazole trifluoroacetate (T29): The substituted 4-hydroxybenzonitrile used was 3-methyl-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromododecane. T29 is a white solid powder with a yield of 86%. 1 H NMR (800 MHz, Methanol- d4) δ 7.83 (d, J = 8.5 Hz, 1H), 7.79 (s, 1H), 6.98 (d, J = 8.6 Hz, 1H), 4.31 – 4.26 (m, 2H), 4.10 – 4.03 (m, 4H), 3.52 (p, J = 8.1 Hz, 1H), 3.35 (d, J = 7.6 Hz, 2H), 2.25 (s, 3H), 1.82 (p, J = 6.7 Hz, 2H), 1.52 (p, J = 7.5 Hz, 2H), 1.40 (p, J = 7.6, 7.1 Hz, 2H), 1.37 – 1.25 (m, 14H), 0.89 (t, J = 7.1 Hz, 3H). 13 C NMR (200 MHz, Methanol- d 4) δ 176.76, 168.01, 161.71 (q, J = 34.8, 34.2 Hz, 1C), 159.85, 128.94, 127.03, 126.37, 117.97, 116.95 (d, J = 283.8 Hz, 1C), 110.59, 67.82, 50.73 (2C), 31.67, 29.41, 29.36 (2C), 29.29, 29.28, 29.08, 29.03, 28.91, 28.54, 25.82, 22.34, 15.00, 13.04. IR (KBr, cm -1 ): 645.94, 720.69, 753.66, 792.02, 824.38, 864.73, 910.94, 1017.14, 1134.57, 1171.74, 1199.80, 1262.34, 1306.04, 1340.68, 1371.21, 1394.74, 1428.70, 1468.83, 1494.37, 1574.39, 1605.81, 1695.77, 2678.22, 2854.54, 2921.39. HRMS (ESI): m / z calcd for C 25 H 39 N3O2[M+H] + : 414.3121 found: 414.3085. Example 5: 5-(azetidin-3-ylmethyl)-3-(3-methyl-4-(nona-8-en-1-yloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (T30): The substituted p-hydroxybenzonitrile used was 3-methyl-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromononene. T30 is a white solid powder with a yield of 88%. 1 H NMR (800 MHz, Methanol- d 4) δ 7.83 (d, J = 8.5 Hz, 1H),7.79 (s, 1H), 6.98 (d, J = 8.6 Hz, 1H), 5.81 (ddt, J = 17.1, 10.1, 6.8 Hz,1H), 4.98 (dd, J = 17.1, 1.9 Hz, 1H), 4.91 (d, J = 9.1 Hz, 1H), 4.30 – 4.26(m, 2H), 4.10 – 4.03 (m, 4H), 3.53 (dt, J = 15.1, 7.6 Hz, 1H), 3.35 (d, J =7.6 Hz, 2H), 2.24 (s, 3H), 2.06 (q, J = 6.3 Hz, 2H), 1.83 (p, J = 6.7 Hz,2H), 1.52 (p, J = 7.3 Hz, 2H), 1.44 – 1.34 (m, 6H). 13 C NMR (200 MHz, Methanol- d 4) δ 176.76, 168.01, 161.68 (q, J = 34.0 Hz, 1C), 159.85, 138.69, 128.94,127.04, 126.37, 117.97, 116.16, 113.35, 110.60, 67.81, 50.73(2C), 33.46,29.41, 28.89, 28.88, 28.74, 28.62, 28.53, 25.79, 14.98. IR (KBr, cm -1): 721.18, 824.49, 909.17, 1017.36, 1135.61, 1171.31, 1199.45, 1261.30, 1306.89, 1340.78, 1370.62, 1394.80, 1428.14, 1496.10, 1573.98, 1608.67, 1694.17, 2698.46, 2858.94, 2925.50, 3525.53. HRMS (ESI): m / z calcd for C 22 H 31 N3O2[M+H] + : 370.2495 found: 370.2459. Example 6: 5-(azetidin-3-ylmethyl)-3-(3-methyl-4-(pent-4-en-1-yloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (T31): The substituted 4-hydroxybenzonitrile used was 3-methyl-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromopentene. White solid, yield 87% 1 H NMR (800 MHz, Methanol- d 4) δ 7.84 (d, J = 2.3 Hz, 1H), 7.80 (s, 1H), 6.98 (d, J = 8.5 Hz, 1H), 5.89 (ddt, J = 17.0, 10.2, 6.7 Hz, 1H), 5.06 (dq, J = 17.1, 1.7 Hz, 1H), 4.99 (d, J = 10.2 Hz, 1H), 4.29 (dd, J = 11.7, 8.8 Hz, 2H), 4.11 – 4.05 (m, 4H), 3.55 – 3.50 (m, 1H), 3.35 (d, J = 7.6 Hz, 2H), 2.31 – 2.27 (m, 2H), 2.26 (s, 3H), 1.92 (dt, J = 13.4, 6.3 Hz, 2H). 13 C NMR (200 MHz, Methanol- d4) δ 176.78, 168.00, 161.71 (d, J = 34.1 Hz, 1C), 159.76, 137.67, 128.95, 127.06, 126.37, 118.05, 116.86 (d, J = 284.2 Hz, 1C), 114.22, 110.61, 67.02, 50.73(2C), 29.93, 29.41, 28.53, 28.23, 14.97. IR (KBr,cm -1 ): 433.02, 647.44, 718.57, 825.53, 912.84, 1030.83, 1135.09, 1172.70, 1199.69, 1257.80, 1307.27, 1340.23, 1394.48, 1429.58, 1493.05, 1573.55, 1607.85, 1695.49, 2693.62, 2954.93. HRMS (ESI): m / z calcd for C 18 H 23 N3O2[M+H] + : 314.1869 found: 314.1834. Example 7: 5-(azetidin-3-ylmethyl)-3-(3-methyl-4-(nonyloxy)phenyl)-1,2,4-oxadiazole trifluoroacetate (T32): The substituted 4-hydroxybenzonitrile used was 3-methyl-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromononane. White solid, yield 88%, 1 H NMR (800 MHz, Methanol- d 4) δ 7.83 (dd, J = 8.5, 2.3 Hz, 1H), 7.79(s, 1H), 6.98 (d, J = 8.5 Hz, 1H), 4.30 – 4.26 (m, 2H), 4.09 – 4.04 (m, 4H), 3.55 – 3.50 (m, 1H), 3.35 (d, J = 7.7 Hz, 2H), 2.25 (s, 3H), 1.85 – 1.80 (m, 2H), 1.54 – 1.50 (m, 2H), 1.40 (p, J= 7.0 Hz, 2H), 1.37 – 1.27 (m, 8H), 0.90(t, J = 7.1 Hz, 3H). 13 C NMR (200 MHz, Methanol- d 4) δ 176.76, 168.01, 161.54 (d, J = 34.4 Hz, 1C), 159.86, 128.93, 127.04, 126.37, 117.96, 116.85 (d, J = 293.0Hz, 1C), 110.60, 67.83, 50.73(2C), 31.64, 29.41, 29.27, 29.07, 28.98, 28.92,28.53, 25.84, 22.33, 14.98, 13.03. IR (KBr, cm -1 ): 720.66, 824.66, 908.59,1015.73, 1135.33, 1172.05, 1199.51, 1261.08, 1308.67, 1340.73, 1370.07,1394.78, 1428.07, 1497.91, 1572.94, 1607.96, 1693.13, 2857.14, 2923.59,2957.30. HRMS (ESI): m / z calcd for C 22 H 33 N3O2[M+H] + : 372.2651 found: 372.2614. Example 8: 5-(azetidin-3-ylmethyl)-3-(4-(decyloxy)-3-methylphenyl)-1,2,4-oxadiazole trifluoroacetate (T33): The substituted 4-hydroxybenzonitrile used was 3-methyl-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromododecane. White solid, yield 89%, 1 H NMR (800 MHz, Methanol- d 4) δ 7.83 (dd, J = 8.4, 2.2 Hz, 1H), 7.79(d, J = 2.2 Hz, 1H), 6.98 (d, J= 8.5 Hz, 1H), 5.80 (ddt, J = 17.0, 10.2, 6.7 Hz,1H), 4.97 (dq, J = 17.1, 1.8 Hz, 1H), 4.91 (dd, J = 10.3, 1.9 Hz, 1H), 4.87 (s,2H), 4.32 – 4.26 (m, 2H), 4.10 – 4.06 (m, 2H), 4.05 (t, J = 6.3 Hz, 2H), 3.53(hept, J = 8.0 Hz, 1H), 3.35 (d, J = 7.6 Hz, 2H), 2.24 (s, 3H), 2.04 (q, J = 7.1Hz, 2H), 1.82 (dt, J = 14.9, 6.5 Hz, 2H), 1.52 (p, J = 7.4 Hz, 2H), 1.44 – 1.30(m, 8H). 13 C NMR (200 MHz, Methanol- d 4) δ 176.77, 168.01, 161.73 (q, J = 34.5 Hz,1C), 159.85, 138.72, 128.94, 127.03, 126.37, 117.97, 113.32, 110.60, 67.82,50.72, 33.49, 29.41, 29.12, 29.01, 28.90, 28.71 (d, J = 7.4 Hz, 4C), 28.54,25.81, 14.99. IR (KBr, cm -1 ): 654.80, 721.51, 824.87, 909.23, 1017.59,1135.82, 1171.54, 1199.34, 1261.65, 1306.49, 1340.65, 1369.40, 1394.94,1428.47, 1468.86, 1495.17, 1574.45, 1608.01, 1693.92, 2697.84, 2857.48,2925.20. HRMS (ESI): m / z calcd for C23 H 35 N3O2[M+H] + : 386.2808 found: 386.2808. Example 9: 5-(Azetidin-3-ylmethyl)-3-(4-(heptyloxy)-3-methylphenyl)-1,2,4-oxadiazole trifluoroacetate (T35): The substituted p-hydroxybenzonitrile used was 3-methyl-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-chloroheptane. White solid, yield 86%. 1 H NMR (800 MHz, Methanol- d 4) δ 7.83 (dd, J J = 8.5, 2.2 Hz, 1H), 7.79(s, 1H), 6.98 (d, J J = 8.5 Hz, 1H), 4.29 (dd, J J = 11.7, 8.8 Hz, 2H), 4.10 – 4.06(m, 2H), 4.05 (t, J J = 6.3 Hz, 2H), 3.55 – 3.50 (m, 1H), 3.35 (d, J J = 7.6 Hz, 2H),2.24 (s, 3H), 1.85 – 1.80 (m, 2H), 1.52 (p, J J = 7.5 Hz, 2H), 1.40 (p, J J = 6.9 Hz,2H), 1.37 – 1.32 (m, 4H), 0.92 (t, J J = 6.9 Hz, 3H). 13 C NMR (200 MHz, Methanol- d 4) δ 176.77, 168.01, 161.73 (q, J J = 34.4 Hz, 1C), 159.85, 128.93, 127.03,126.37, 117.97, 116.85 (d, J J = 293.1 Hz, 1C), 110.59, 67.83, 50.72(2C), 31.59,29.41, 28.94, 28.78, 28.53, 25.83, 22.27, 14.97, 13.02. IR (KBr, cm -1): 647.98, 721.08, 751.65, 823.08, 909.00, 1037.68, 1131.82, 1197.52, 1255.34, 1308.29, 1393.18, 1429.32, 1467.51, 1574.70, 1607.54, 1707.26, 2673.33, 2860.83, 2926.91. HRMS (ESI): m / z calcd for C 20 H 29 N3O2[M+H] + : 344.2338 found: 344.2301. Example 10: 5-(azetidin-3-ylmethyl)-3-(4-(nonyloxy)-3-(trifluoromethyl)phenyl)-1,2,4-oxadiazole trifluoroacetate (T40): The substituted p-hydroxybenzonitrile used was 3-trifluoromethyl-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromononane. White solid, yield 89%. 1 H NMR (800 MHz, Chloroform- d ) δ 8.22 (d, J J = 2.2 Hz, 1H), 8.13 (dd, J J = 8.7, 2.2 Hz, 1H), 7.05 (d, J J = 8.8 Hz, 1H), 4.33 (s, 2H), 4.10 (t, J J = 6.4 Hz, 2H), 4.08 – 4.04 (m, 2H), 3.52 (h, J J = 7.9 Hz, 1H), 3.35 (d, J J = 7.8 Hz, 2H), 1.83 (dq, J J = 14.3, 7.8, 7.2 Hz, 2H), 1.48 (p, J J = 7.4 Hz, 2H), 1.35 (q, J J = 7.9 Hz, 2H), 1.33 – 1.24 (m, 8H), 0.88 (t, J J = 7.1 Hz, 3H). 13 C NMR (200 MHz, Chloroform- d) δ 176.08, 167.35, 162.56 (d, J = 39.4 Hz, 1C), 159.40, 132.41,126.58 (q, J = 5.2 Hz, 1C), 123.18 (d, J = 272.6 Hz, 1C), 119.45 (q, J = 31.5 Hz,1C), 117.96,116.50 (d, J = 291.6 Hz, 1C), 112.94, 69.09, 50.50(2C), 31.86,29.46, 29.38, 29.34, 29.22, 29.21, 28.88, 25.74, 22.67, 14.10. IR (KBr, cm -1 ):417.28, 676.10, 721.26, 832.76, 916.04, 1003.03, 1056.38, 1136.45, 1202.72,1286.58, 1319.47, 1351.22, 1427.84, 1474.56, 1624.53, 1696.14, 2857.81,2926.00, 2968.68, 3441.82. HRMS (ESI): m / z calcd for C 22 H 30 F3N3O2[M+H] + : 426.2368found: 426.2349. Example 11: 3-(3-Bromo-4-(nonyloxy)phenyl)-5-(pyrrolidin-3-ylmethyl)-1,2,4-oxadiazole trifluoroacetate (T42): The substituted p-hydroxybenzonitrile 3-bromo-4-hydroxybenzonitrile, hydroxylamine hydrochloride, and sodium bicarbonate were dissolved in methanol and refluxed for 22 hours. The reaction solution was cooled to room temperature and filtered by suction, and the solvent was removed under reduced pressure to obtain a crude product of aminooxime. The obtained crude product, N -Boc-3-pyrrolidineacetic acid, DIPEA were dissolved in DMF, and HATU was added, and the reaction was carried out at 100 o °C for 12 hours. The reaction solution was extracted with ethyl acetate, and the crude product was separated and purified by silica gel column chromatography. Then, this intermediate, cesium carbonate, and the halogenated hydrocarbon 1-bromononane were dissolved in DMF. 75 oReact for 8 hours. Extract the reaction solution with ethyl acetate. Wash the organic phase successively with water and saturated brine, dry over anhydrous sodium sulfate, filter, evaporate the solvent under reduced pressure, and separate and purify the crude product by silica gel column chromatography to obtain the key intermediate. Dissolve the key intermediate in CH2Cl2, and slowly add dropwise TFA with stirring, and react at room temperature for 4 hours. After the reaction is completed, evaporate the solvent under reduced pressure, and separate and purify the crude product by silica gel column chromatography (methylene chloride - methanol = 15:1) to obtain the white target compound T42 with a yield of 85%.

[0026] T42 is a colorless oily liquid with a yield of 85%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.12 (s, 1H),7.97 (d, J = 8.6 Hz, 1H), 7.29 (d, J = 8.7 Hz, 1H), 4.14 (t, J = 6.4 Hz, 2H), 3.18(t, J = 8.1 Hz, 3H), 2.96 – 2.90 (m, 1H), 2.75 (p, J = 7.7 Hz, 1H), 2.17 (dt, J =13.0, 6.4 Hz, 1H), 1.77 (p, J = 7.0 Hz, 2H), 1.71 – 1.62 (m, 1H), 1.46 (t, J =7.6 Hz, 2H), 1.31 (d, J = 35.9 Hz, 12H), 0.88 – 0.83 (m, 3H). 13 C NMR (100 MHz,DMSO- d 6) δ 179.24, 166.70, 162.39, 157.73, 131.71, 128.46, 120.07, 116.13,114.50, 112.02, 69.46, 49.75, 45.05, 35.54, 31.72, 30.19, 29.39, 29.08,29.06, 28.97, 28.83, 25.83, 22.56, 14.40. IR (KBr, cm -1): 480.07, 504.16, 583.98, 638.65, 711.93, 800.04, 826.48, 901.20, 1009.88, 1046.34, 1129.73, 1197.63, 1272.62, 1349.90, 1392.96, 1465.28, 1522.68, 1594.30, 1673.31, 2856.89, 2924.81. HRMS (ESI): m / z calcd for C 22 H 32 BrN3O2[M+H] + : 450.1756 found: 450.1717 The preparation methods of the compounds in Examples 12 - 17 are the same as those in Example 11, except that different substituted 4 - hydroxybenzonitriles and halogenated hydrocarbons are used. The substituted 4 - hydroxybenzonitriles and halogenated hydrocarbons used in each specific example are indicated in the corresponding example: Example 12: 3 - (4 - ((3,5 - bis(trifluoromethyl)benzyl)oxy) - 3 - bromophenyl) - 5 - (pyrrolidin - 3 - ylmethyl) - 1,2,4 - oxadiazole trifluoroacetate (T44): The substituted 4 - hydroxybenzonitrile is 3 - bromo - 4 - hydroxybenzonitrile, and the halogenated hydrocarbon used is 1 - (bromomethyl) - 3,5 - bis(trifluoromethyl)benzene. T44 is a white solid with a yield of 87%. 1 H NMR (400 MHz, DMSO - d 6) δ 8.23 (s, 2H), 8.19(d, J = 2.1 Hz, 1H), 8.13 (s, 1H), 8.04 (dd, J = 8.5, 2.1 Hz, 1H), 7.43 (d, J = 8.7Hz, 1H), 5.53 (s, 2H), 3.32 (s, 2H), 3.20 (t, J = 9.1 Hz, 3H), 2.99 (t, J = 9.3Hz, 1H), 2.79 (p, J = 7.8 Hz, 1H), 2.20 (dq, J = 12.8, 6.9 Hz, 1H), 1.71 (dq, J =17.1, 8.7 Hz, 1H). 1313C NMR (100 MHz, DMSO- d 6) δ 179.25, 166.60, 160.51, 156.85,140.30, 131.90, 130.89 (q, J J = 32.8 Hz, 2C), 128.52 (d, J J = 10.9 Hz, 4C), 125.11,122.39, 121.01, 119.73, 115.09, 112.25, 69.23, 49.42, 44.88, 35.38, 29.99,28.80. IR (KBr, cm -1 ): 417.24, 441.05, 679.01, 717.79, 802.02, 838.70, 890.99,1132.66, 1175.15, 1203.78, 1279.05, 1365.47, 1397.93, 1426.83, 1452.32,1480.94, 1593.30, 1682.47, 2990.87, 3441.86. HRMS (ESI): m / z calcd forC 22 H 18 BrF6N3O2[M+H] + :550.0565 found: 550.0538. Example 13: (R)-3-(3-Bromo-4-(nonyloxy)phenyl)-5-(pyrrolidin-3-ylmethyl)-1,2,4-oxadiazole trifluoroacetate (T47(R)): The substituted p-hydroxybenzonitrile was 3-bromo-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromononane. T47(R) was a colorless oily liquid with a yield of 86%. +37.22 ( c J = 0.04, MeOH). 1 1H NMR (800 MHz, Methanol- d 4) δ 8.20– 8.16 (m, 1H), 8.00 – 7.95 (m, 1H), 7.14 (q, J J = 9.1, 8.0 Hz, 1H), 4.12 (p, J J =6.4 Hz, 2H), 3.65 (dd, J= 11.8, 7.7 Hz, 1H), 3.46 (ddd, J = 12.1, 8.3, 4.1 Hz,1H), 3.34 – 3.32 (m, 1H), 3.24 – 3.16 (m, 2H), 3.11 (dd, J = 12.0, 8.9 Hz, 1H),2.94 (hept, J = 7.4 Hz, 1H), 2.37 (dtd, J = 14.4, 7.4, 4.2 Hz, 1H), 1.85 (dq, J =13.0, 8.9 Hz, 3H), 1.54 (d, J = 7.5 Hz, 2H), 1.40 (s, 2H), 1.31 (d, J = 31.9 Hz,8H), 0.89 (ddd, J = 7.2, 4.3, 2.6 Hz, 3H). 13 C NMR (200 MHz, Methanol- d 4) δ178.13, 166.88, 161.71 (d, J = 36.1 Hz, 1C), 157.91, 131.66, 127.65, 119.87,116.70 (d, J = 290.2 Hz, 1C), 112.89, 111.86, 69.05(2C), 49.32, 44.91, 35.37,31.64, 29.65, 29.26, 28.97, 28.70, 28.24, 25.70, 22.35, 13.06. IR (KBr, cm -1 ):440.29, 472.02, 492.85, 520.71, 599.60, 639.42, 719.76, 799.92, 839.43,900.38, 1049.46, 1132.04, 1193.46, 1271.07, 1336.38, 1391.47, 1462.50,1592.68, 1673.63, 2855.52, 2924.67. HRMS (ESI): m / z calcd for C 22 H 32 BrN3O2[M+H] +: 450.1756 found: 450.1708. Example 14: (R)-3-(3-Bromo-4-(heptyloxy)phenyl)-5-(pyrrolidin-3-ylmethyl)-1,2,4-oxadiazole trifluoroacetate (T48(R)): The substituted p-hydroxybenzonitrile was 3-bromo-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-bromoheptane. T48 was a colorless oily liquid with a yield of 86%. +22.54 ( c = 0.04, MeOH). 1 1H NMR (800 MHz, Methanol- d 4) δ 8.20– 8.17 (m, 1H), 7.98 (dd, J = 8.6, 2.1 Hz, 1H), 7.13 (d, J = 8.6 Hz, 1H), 4.12(t, J = 6.3 Hz, 2H), 3.66 (dd, J = 11.8, 7.7 Hz, 1H), 3.46 (ddd, J = 12.3, 8.5, 4.3Hz, 1H), 3.20 (qd, J = 16.2, 7.3 Hz, 2H), 3.12 (dd, J = 11.8, 9.0 Hz, 1H), 2.94(hept, J = 7.4 Hz, 1H), 2.40 – 2.35 (m, 1H), 1.86 (dq, J = 13.3, 8.9 Hz, 3H),1.57 – 1.51 (m, 2H), 1.41 (s, 2H), 1.34 (d, J = 3.6 Hz, 5H), 0.91 (dt, J = 15.2,7.0 Hz, 3H). 13 13C NMR (200 MHz, Methanol- d 4) δ 178.13, 166.88, 116.73 (d, J= 290.6 Hz, 1C), 157.90, 131.66, 127.66, 119.87, 116.01, 112.88, 111.85, 69.06, 49.32, 44.92, 35.37, 31.57, 29.65, 28.72, 28.70, 28.24, 25.70, 22.26, 13.03. IR (KBr, cm -1 ): 424.71, 439.25, 470.86, 493.38, 520.93, 599.61, 639.21, 719.86, 799.89, 839.68, 900.11, 1013.31, 1049.40, 1131.83, 1193.00, 1271.15, 1336.50, 1391.64, 1462.27, 1592.84, 1673.53, 2849.11, 2926.29. HRMS (ESI): m / z calcd for C 20 H 28 BrN3O2[M + H] + : 422.1443 found: 422.1400. Example 15: (S)-3-(3-Bromo-4-(heptyloxy)phenyl)-5-(pyrrolidin-3-ylmethyl)-1,2,4-oxadiazole trifluoroacetate (T54): The substituted p-hydroxybenzonitrile was 3-bromo-4-hydroxybenzonitrile, and the halogenated hydrocarbon used was 1-chloroheptane. T54 was a colorless oily liquid with a yield of 89%. -23.81 ( c = 0.04, MeOH). 1 1H NMR (800 MHz, Methanol- d 4) δ 8.22 (d, J = 2.0 Hz, 1H), 8.01 (dd, J = 8.6, 2.1 Hz, 1H), 7.17 (d, J = 8.6 Hz, 1H), 4.15 (t, J = 6.3 Hz, 2H), 3.66 (dd, J = 7.9, 3.9 Hz, 1H), 3.47 (ddd, J= 12.4, 8.5, 4.3 Hz, 1H), 3.22 (qd, J = 16.3, 7.3 Hz, 2H), 3.12 (dd, J = 11.8, 8.9 Hz, 1H), 2.96 (p, J = 8.0 Hz, 1H), 2.41 – 2.37 (m, 1H), 1.87 (dd, J = 14.5, 6.8 Hz, 2H), 1.57 (p, J = 7.6 Hz, 2H), 1.46 – 1.29 (m, 8H), 0.94 (t, J = 6.8 Hz, 3H). 13 C NMR (200 MHz, Methanol- d 4) δ 178.14, 166.90, 161.67 (d, J = 47.2 Hz, 1C), 157.94, 131.68, 127.63, 119.87, 116.85 (d, J = 293.2 Hz, 1C), 112.90, 111.85, 69.07, 49.37, 44.96, 35.38, 31.57, 29.64, 28.71, 28.69, 28.23, 25.70, 22.25, 13.02. IR (KBr, cm -1 ): 722.82, 835.78, 902.85, 1014.34, 1050.45, 1133.31, 1206.20, 1273.49, 1338.26, 1421.88, 1465.65, 1559.02, 1597.01, 1681.88, 2858.95, 2928.07. HRMS (ESI): m / z calcd for C 20 H 28 BrN3O2 [M + H] + : 422.1443 found: 422.1396. Example 16: (S)-3-(3-Bromo-4-(nonyloxy)phenyl)-5-(pyrrolidin-3-ylmethyl)-1,2,4-oxadiazole trifluoroacetate (T55(S)): The substituted p-hydroxybenzonitrile is 3-bromo-4-hydroxybenzonitrile, and the halogenated hydrocarbon used is 1-bromononane. T55 is a white solid with a yield of 90%. -32.61 ( c = 0.04, MeOH), m.p: 63 - 65 °C. 1 1H NMR (800 MHz, Methanol- d 4) δ 8.21 (d, J = 2.1 Hz, 1H), 8.01 (dd, J = 8.6, 2.1 Hz, 1H), 7.17 (d, J = 8.6Hz, 1H), 4.15 (t, J = 6.2 Hz, 2H), 3.67 (dd, J = 11.8, 7.7 Hz, 1H), 3.48 (ddd, J =12.4, 8.5, 4.3 Hz, 1H), 3.22 (qd, J = 16.2, 7.3 Hz, 3H), 3.13 (dd, J = 11.8, 8.9Hz, 1H), 2.96 (p, J = 7.4 Hz, 1H), 2.42 – 2.37 (m, 1H), 1.57 (p, J = 7.5 Hz, 2H),1.45 – 1.29 (m, 13H), 0.92 (t, J = 7.0 Hz, 3H). 13 13C NMR (200 MHz, Methanol- d 4) δ178.13, 166.89, 161.60 (d, J = 34.1 Hz, 1C), 157.92, 131.67, 127.64, 119.87,116.82 (d, J = 292.6 Hz, 1C), 112.90, 111.86, 69.06, 49.34, 44.94, 35.37,31.64, 29.63, 29.25, 28.98, 28.96, 28.69, 28.22, 25.70, 22.34, 13.04. IR(KBr, cm -1): 718.58, 747.77, 795.65, 828.52, 901.78, 1008.65, 1048.11, 1129.77, 1167.35, 1210.21, 1271.91, 1357.32, 1395.78, 1467.60, 1592.76, 1674.91, 2778.79, 2854.75, 2926.88. HRMS (ESI): m / z calcd for C 22 H 32 BrN3O2[M+H] + : 450.1756 found: 450.1704. Example 17: (S)-3-(3-Bromo-4-((3-(trifluoromethyl)benzyl)oxy)phenyl)-5-(pyrrolidin-3-ylmethyl)-1,2,4-oxadiazole trifluoroacetate (T57(S)): The substituted p-hydroxybenzonitrile is 3-bromo-4-hydroxybenzonitrile, and the halogenated hydrocarbon used is 1-(chloromethyl)-3-(trifluoromethyl)benzene. T57(S) is a white solid with a yield of 87%. -32 ( c = 0.04, MeOH), m.p: 105 - 107 °C. 1 H NMR(800 MHz, Methanol- d 4) δ 8.14 (d, J = 2.0 Hz, 1H), 7.91 (dd, J = 8.6, 2.1 Hz,1H), 7.75 (s, 1H), 7.68 (d, J = 7.6 Hz, 1H), 7.55 (d, J = 7.8 Hz, 1H), 7.51 (t, J =7.7 Hz, 1H), 7.17 (d, J = 8.6 Hz, 1H), 5.25 (s, 2H), 3.56 (dd, J = 11.8, 7.8 Hz,1H), 3.36 (ddd, J = 12.4, 8.5, 4.3 Hz, 1H), 3.25 – 3.22 (m, 1H), 3.10 (qd, J =16.2, 7.3 Hz, 2H), 3.01 (dd,J = 11.8, 8.9 Hz, 1H), 2.87 – 2.82 (m, 1H), 2.30 –2.25 (m, 1H), 1.76 (dq, J = 13.2, 8.9 Hz, 1H). 13 C NMR (200 MHz, Methanol- d 4) δ 1178.23, 166.78, 161.65 (q, J = 34.9 Hz, 1C), 157.11, 137.78, 131.83, 130.54 (q, J = 32.2 Hz, 1C), 129.07, 127.67, 124.41 (q, J = 3.9 Hz, 2C), 123.43 (q, J = 3.9Hz, 2C), 120.74, 116.86 (d, J = 293.4 Hz, 1C), 113.64, 112.11, 69.62, 49.32,44.93, 35.36, 29.63, 28.23. IR (KBr, cm -1 ): 715.35, 747.44, 802.82, 902.36,1059.96, 1127.44, 1170.89, 1205.67, 1275.40, 1331.67, 1397.48, 1448.67,1484.21, 1532.72, 1591.08, 1676.25, 2800.71, 3003.02. HRMS (ESI): m / z calcd forC 21 H 19 BrF3N3O2[M+H] + :482.0691 found: 482.0672. The partial pharmacological activity results of the compounds of the present invention are as follows: Experimental Example 1: Inhibition rate and IC 50 activity of the compounds of the present invention against SphK1 and SphK2 1. Experimental method Dissolve the compound in DMSO to prepare a 10 mmol stock solution, and perform serial dilutions with kinase buffer (pH = 7.4, composition: 40 mmol / L Tris, 10 mmol / L MgCl2, 0.1 g / L BSA, 1 mmol / L DTT, 10 µM / L ATP). The concentration at the initial screening is set to 10 µM. When measuring the IC 50 value, set a total of 5 concentration gradients: 0.01 µM, 0.1 µM, 1 µM, 10 µM, and 100 µM.

[0027] On a 96-well microplate, first add the drug, then add the kinase buffer and mix well, then add the Sphingosine substrate (0.2 mM), and finally add SphK1 or SphK2 and mix well. Incubate at 30 ºC for 40 min. Then add the ATP test solution, and incubate the mixture at room temperature for 5 min. Immediately measure the luminescence signal (Luminescence, Lu) using a microplate spectrophotometer. Substitute the values into the following formula to calculate the inhibition rate: % activity = [(Lu drug - Lu background) / (Lu enzyme - Lu background)] × 100%.

[0028] Process with GraphPad Prism 10 software to calculate the IC 50 value.

[0029] 2. The experimental results are shown in Table 1 below.

[0030] Table 1 Inhibition rates (%) of the target compounds (10 μM) on SphK1 / 2 Note: a IC 50 value, b ND means not tested, PF-543 is the positive control drug for SphK1, and K145 is the positive control drug for SphK2.

[0031] When tested at 10 μM, most of the 17 compounds showed inhibition rates of over 40% and even up to over 96% on SphK1 and SphK2. Among them, T16, T18, T23, T29, T30, T31, T32, T33, T35, T42, T44, T47(R), T48(R), T55(S), T57(S) showed strong inhibitory effects on both phK1 and SphK2.

[0032] Based on the inhibitory effects of the target compounds on the activities of phK1 and SphK2 at a concentration of 10 μM, we selected some compounds with relatively ideal activity results and tested their IC on phK1 and SphK250 , the test results are shown in Table 2: Table 2 IC of the compound against SphKs 50 Note: a PF-543 is the positive control drug for SphK1, and K145 is the positive control drug for SphK2.

[0033] The activity results show that the compounds involved in the present invention have ideal inhibitory effects on both SPhK1 and SPhK2. Therefore, the anti-proliferation effects of some compounds on tumor cells were further determined.

[0034] Experimental Example 2: Evaluation of the anti-proliferation effects of compounds on tumor cells (A549 cells, HCT116 cells, MDA-MB-231 cells) 1. Experimental method The tumor cells (A549 cells, HCT116 cells, MDA-MB-231 cells) were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, and subculture was performed every 2 to 3 days. The cell culture incubator was maintained at 37 °C and 5% CO2, and cells in the logarithmic growth phase were used for the experiment.

[0035] The suspension of tumor cells (6×103 / well / 100 μL) was added to a 96-well plate and cultured for 24 h. Different concentrations of the test compounds (0.1 μM, 0.3 μM, 1 μM, 3 μM, 10 μM, 30 μM, and 100 μM) and cisplatin were added respectively, and the culture was continued for 48 h. After the culture was completed, the supernatant was removed, 100 μL of medium containing 0.5 mg / mL CCK-8 was added to each well, and the culture was continued for 2.5 h under the conditions of 37 °C and 5% CO2. The absorbance (OD value) of each well at 450 nm was measured by a multifunctional microplate reader. The cell survival rate was calculated according to the formula "Cell survival rate (%) = (OD value of the treatment group / OD value of the control group) × 100%", and the obtained data was analyzed by GraphPad Prism 10 software.

[0036] 2. Experimental results Table 3 Anti-proliferation effects of compounds on tumor cells Note: Cisplatin is the positive control drug.

[0037] The activity results (Table 3) showed that in the inhibitory activity against A549 cells, all the compounds exhibited better inhibitory effects than cisplatin. In particular, T16, T33, T42, T47(R) and T55(S) had the best activities, with IC50 values of 4.12±1.41 μM, 4.78±0.18 μM, 4.60±0.61 μM, 3.27±0.25 μM, and 4.56±0.45 μM, respectively. In the inhibitory activity against HCT116 cells, the activities of T16, T18, T30, T32, T33, T42, T47(R), T48(R), T55(S), and T57(S) were all better than that of cisplatin, with IC50 values of 5.37±0.62 μM, 5.27±0.22 μM, 4.32±0.11 μM, 4.29±0.11 μM, 4.13±0.21 μM, 4.14±0.26 μM, 3.87±0.06 μM, 5.43±0.23 μM, 4.11±0.21 μM, and 3.74±0.14 μM, respectively. In the inhibitory activity against MDA-MB-231 cells, the activities of all the compounds were better than that of cisplatin. Among them, the activities of T16 and T47(R) were particularly significant, with IC50 values of 4.47±0.14 μM and 4.00±0.07 μM, respectively.

Claims

1. 1,2,4-oxadiazole compounds containing nitrogen heterocycles represented by general formula (I) or their solvates: Formula (I) Among them, n = 1, 2, R 1 = F, Br, CH3, CF3, R 2 is 。 2. The 1,2,4-oxadiazole compound containing a nitrogen heterocycle or its solvate according to claim 1, characterized in that, The structural formula of the said compound is: 。 3. The 1,2,4-oxadiazole compound containing a nitrogen heterocycle or its solvate according to claim 2, characterized in that, The structural formula of the said compound is: 。 4. A pharmaceutical composition, which contains the 1,2,4-oxadiazole compound containing nitrogen heterocycle or its solvate according to any one of claims 1-3 and a pharmaceutically acceptable carrier.

5. Use of the 1,2,4-oxadiazole compound containing nitrogen heterocycle or its solvate according to any one of claims 1-3 or the pharmaceutical composition according to claim 3 in the preparation of a drug for preventing and / or treating diseases mediated by SphK1 and / or SphK2.

6. The application according to claim 5, characterized in that, The diseases mediated by SphK1 are cancer and inflammatory diseases.

7. The application according to claim 6, characterized in that, The cancer includes colon cancer, lung cancer, breast cancer, liver cancer, gastric cancer, lung adenocarcinoma, melanoma, esophageal cancer; the inflammatory diseases include inflammatory bowel disease, hepatitis, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis or multiple sclerosis.

8. The application according to claim 5, wherein The diseases mediated by SphK2 are cancer, inflammatory diseases, ischemia-reperfusion injury, renal fibrosis, Alzheimer's disease, diabetic retinopathy, diabetic nephropathy.

9. The application according to claim 8, wherein The cancer is rectal cancer, gastric cancer, renal cancer, colon cancer, uterine cancer, ovarian cancer, breast cancer, lung cancer.

10. The application according to claim 9, characterized in that, The cancer is lung cancer, colon cancer, breast cancer.

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