Epimedium-derived icariside ii and preparation method and application thereof with anti-hepatoma activity

Abstract

The application belongs to the technical field of medicine, and particularly relates to icariin II derivative with anti-hepatoma activity and a preparation method and application thereof. The application prepares a series of icariin II (ICA-II) derivatives with high anti-hepatoma activity, and further provides a pharmaceutical composition containing the derivative or a pharmaceutically acceptable salt thereof, and a preparation method of the derivative or the pharmaceutically acceptable salt thereof. In addition, the application also provides the use of the derivative or the pharmaceutically acceptable salt thereof or the pharmaceutical composition thereof in the preparation of a drug for treating primary hepatocarcinoma.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to icariin II derivatives with anti-hepatocellular carcinoma activity, their preparation methods, and applications. Background Technology

[0002] Liver cancer is one of the most common cancers worldwide and a leading cause of cancer-related deaths. Hepatocellular carcinoma (HCC) is the most common form of liver cancer, accounting for approximately 90% of all cases. Causes of liver cancer include (e.g., hepatitis B and C) viral infections, alcohol, obesity, and dietary carcinogens. Current treatments for liver cancer primarily include surgical resection and liver transplantation, but both have limited effectiveness. Targeted therapies for liver cancer are limited to sorafenib, lenvatinib, regorafenib, ramucirumab, and cabozantinib. While these drugs can prolong patient survival, drug resistance has developed (European Journal of Medicinal Chemistry, 2021, 224:113690). Therefore, there is an urgent need to develop alternative treatment strategies.

[0003] Icaritin (ICT), a major component of the traditional Chinese medicine Epimedium, is an isopentenyl flavonoid compound that has been approved for marketing as a Class I innovative drug (drug name: Acoladin). However, it has poor water solubility and limited anti-hepatocellular carcinoma activity. Based on the previous structure-activity relationship study conducted by our team, we found that (1) the presence of rhamnose at the C3 position significantly enhances its activity; (2) the isopentenyl group is a key group with anti-hepatocellular carcinoma activity; and (3) the introduction of a nitrogen-containing heterocycle at the C7 position can enhance its proliferation inhibition effect. Based on this research, our invention further modifies the C7 position to further improve the anti-hepatocellular carcinoma cell proliferation activity of icariin and its structural analogs.

[0004] 1,3,4-Oxadiazole derivatives have been successfully used as antimitotic drugs to treat cancer. Most antimitotic drugs target microtubules, which are dynamic elements in the cytoskeleton responsible for the formation of the spindle during mitosis and are essential for chromosome segregation during cell division. Oxadiazole derivatives can inhibit microtubule polymerization and prevent tumor cell mitosis, thereby exerting antitumor activity (Bioorganic & Medicinal Chemistry Letters, 2006, 16: 1191-1196). In addition, 1,3,4-oxadiazole and 1,3,4-thiadiazole are also popular fragments in antitumor research. Therefore, the inventors attempted to introduce them to enhance the antiproliferative activity of icariin against liver cancer cells. Summary of the Invention

[0005] To address the aforementioned technical problems, the inventors have prepared a series of icariin II (ICA-II) derivatives with high anti-hepatocellular carcinoma activity, and further provided pharmaceutical compositions comprising the derivatives or pharmaceutically acceptable salts thereof, as well as methods for preparing the derivatives or pharmaceutically acceptable salts thereof. Furthermore, the use of the derivatives or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, in the preparation of medicaments for treating primary liver cancer is also provided.

[0006] Specifically, the present invention is achieved through the following technical solutions:

[0007] In a first aspect, the present invention provides an icariin II derivative or a pharmaceutically acceptable salt thereof with anti-hepatocellular carcinoma activity, wherein the icariin II derivative is an icariin II oxadiazole derivative or an icariin II thiadiazole derivative; wherein the icariin II oxadiazole derivative has a chemical structure shown in formula (I) or formula (II); and the icariin II thiadiazole derivative has a chemical structure shown in formula (III) or formula (IV).

[0008]

[0009] In formulas (I), (II), (III), or (IV), R is selected from:

[0010]

[0011] Preferably, the icariin II oxadiazole derivative or icariin II thiadiazole derivative with anti-hepatocellular carcinoma activity is selected from the following compounds:

[0012]

[0013]

[0014]

[0015]

[0016] In a second aspect, the present invention also provides a method for preparing the icariin II oxadiazole derivative or icariin II thiadiazole derivative with anti-hepatocellular carcinoma activity described in the first aspect, comprising the following steps:

[0017]

[0018] Starting from a1-a22 and a31-a34, intermediates b1-b22 and b31-34 were obtained through step (i), fragments c1-c22 and c31-34 were obtained through step (ii), and finally, icariin II was replaced through step (iii) to obtain compounds YYH1-22 and YYH31-34.

[0019] Starting from a23-a30, intermediate b23-b30 is obtained through step (i), intermediate c23-c30 is obtained through step (iv), fragment d23-30 is obtained through step (v), and finally, icariin II is replaced through step (iii) to obtain compound YYH23-30.

[0020] Specifically, the preparation method includes the following steps:

[0021] (i) Synthesis of b1-b34: Dissolve a1-a34 in solvent, add 1.5-3 equivalents of HOBt and 1.5-3 equivalents of EDCI, stir for 0.5-4 hours, add 2-4 equivalents of 80% hydrazine hydrate dropwise, react at room temperature, remove solvent, extract, wash, dry, filter, and purify.

[0022] (ii) Synthesis of c1-c22 and c31-34: Dissolve b1-b22 and b31-34 in a solvent, add 1-3 equivalents of trimethoxychloroethane and 3-5 equivalents of acetic acid, react at 110-130℃, and the post-treatment is the same as in step (i).

[0023] (iii) Synthesis of YYH1-34: Icariin I and 1-2 equivalents of K2CO3 and 0.5-1 equivalents of KI were dissolved in a solvent, stirred at room temperature, and the corresponding fragment was added. The reaction was carried out at 60°C, and the post-treatment was the same as in step (i).

[0024] (iv) Synthesis of d23-d30: Dissolve b23-b30 in a solvent and add 1-2 equivalents of p-chloroacetic acid, 1.5-3 equivalents of HOBt and 1.5-3 equivalents of EDCI. React at room temperature and the post-treatment is the same as in step (i).

[0025] (v) Synthesis of e23-e30: Dissolve d23-d30 in a solvent, add 2-3 equivalents of Lawson reagent, react at 110-130℃, and the post-treatment is the same as in step (i);

[0026] In the above preparation method, wherein:

[0027] The solvent is selected from anhydrous acetonitrile, N,N-dimethylaminoformamide, 1,4-dioxane, toluene, or acetone.

[0028] In a third aspect, the present invention provides a pharmaceutical composition comprising, in the first aspect, an icariin II oxadiazole derivative or an icariin II thiadiazole derivative having anti-hepatocellular carcinoma activity, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0029] The pharmaceutical composition is available in oral or injectable dosage forms. Oral dosage forms include capsules, tablets, granules, oral liquids, sustained-release formulations, or controlled-release formulations.

[0030] Preferably, the pharmaceutically acceptable carrier is selected from one or more of diluents, lubricants, binders, disintegrants, stabilizers, or solvents.

[0031] More preferably, the diluent of the present invention is selected from starch, microcrystalline cellulose, sucrose, dextrin, lactose, powdered sugar, and glucose; the lubricant is selected from magnesium stearate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, and poloxamer; the binder is selected from water, ethanol, starch paste, syrup, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, and polyvinylpyrrolidone; the disintegrant is selected from starch effervescent mixtures, i.e., sodium bicarbonate and citric acid, tartaric acid, and low-substituted hydroxypropyl cellulose; the stabilizer is selected from polysaccharides such as farnesum gum, agar, alginic acid, cellulose ether, and carboxymethyl chitosan; and the solvent is selected from water and balanced salt solutions.

[0032] In a fourth aspect, the present invention provides the use of the icariin II oxadiazole derivative or icariin II thiadiazole derivative or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the third aspect above, which have anti-hepatocellular carcinoma activity, in the preparation of a medicament for treating primary liver cancer.

[0033] Compared with the prior art, the present invention has the following advantages:

[0034] This invention provides a series of oxadiazole derivatives or thiadiazole derivatives or their pharmaceutically acceptable salts with icariin II as the parent nucleus, providing lead compounds for the preparation of drugs for treating primary liver cancer. Detailed Implementation

[0035] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in the art, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0036] Example 1: Preparation of compound YYH-4:

[0037]

[0038] Take a 25 mL reaction flask and place ICA-II (50 mg, 0.097 mmol), anhydrous potassium carbonate (27 mg, 0.194 mmol), potassium iodide (8 mg, 0.0485 mmol), and 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole (44 mg, 0.194 mmol) into it. Add 3 mL of dry acetone to dissolve the precipitate and stir at 60 °C for 16 h. Monitor the reaction by TLC until complete. Quench the reaction with 2 mL of water, extract three times with 2 mL of ethyl acetate, wash the organic phase with saturated sodium chloride solution, dry with anhydrous sodium sulfate, remove the solvent under reduced pressure, and purify by RP-HPLC to obtain YYH-4, yield: 65.3%. The structural identification data are as follows:

[0039] YYH-4: Yellow solid. 1 H-NMR (600MHz, DMSO-d6): δ12.66 (s, 1H), 7.95 (d, J = 8.9Hz,

[0040] 2H),7.87(d,J=8.8Hz,2H),7.16(d,J=8.9Hz,2H),7.12(d,J=8.8Hz,2H),6.81(s,1H),5.64(s,2H),5.28(s,1H),5.13(t,J=7 .8Hz,1H),5.02(s,1H),4.78(s,2H),4.00(s,1H),3.85(s,6H),3.48(m,3H),3.14(m,2H),1.56(s,6H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.3,162.8,162.0,160.8,159.9,157.9,153.4,135.1,131.9,131.0,129.0(2C),122.7,122.0,115. 7(2C),115.5(2C),114.6(2C),108.2(2C),105.9,102.5,97.0,71.6,71.2,70.8,70.5,60.9,56.0,55.9,25.9,22.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 37 H 39 N2O 12,703.2497; observed 703.2492.

[0041] Example 2: Preparation of compound YYH-23:

[0042]

[0043] Take a 25 mL reaction flask and place ICA-II (50 mg, 0.097 mmol), anhydrous potassium carbonate (27 mg, 0.194 mmol), potassium iodide (8 mg, 0.0485 mmol), and 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-thiadiazole fragment (41 mg, 0.194 mmol) into it. Add 3 mL of dry acetone to dissolve the fragment and stir at 60 °C for 16 h. Monitor the reaction by TLC until complete. Add 2 mL of water to quench the reaction, extract three times with 2 mL of ethyl acetate, wash the organic phase with saturated sodium chloride solution, dry with anhydrous sodium sulfate, remove the solvent under reduced pressure, and purify by RP-HPLC to obtain YYH-23, yield: 34.2%. The structural identification data are as follows:

[0044] YYH-23: Yellow solid. 1 H-NMR (600MHz, DMSO-d6): δ12.66 (s, 1H), 8.06 (m, 2H), 7.87 (d, J = 8.4Hz, 2H), 7 .41(t,J=8.5Hz,2H),7.12(d,J=8.4Hz,2H),6.80(s,1H),5.80(s,2H),5.28(s, 1H),5.14(s,1H),5.00(s,1H),4.74(s,1H),4.68(s,1H),4.00(s,1H),3.85(s, 3H), 3.48 (m, 3H), 3.14 (m, 2H), 1.61 (s, 3H), 1.60 (s, 3H), 0.79 (d, J = 5.7Hz, 3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,168.6,166.2,161.9,160.7,159.9,157.8,153.4,151.9,139.6,135.1,132.0,131.0,130.7,130.6,128.5 ,126.4,125.3,122.6,122.2,117.1,114.5,108.1,105.8,102.5,96.9,71.5,71.1,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS(ESI + ):[M+H] + calcd for C36 H 36 FN2O 11 S,707.2069; observed 707.2059.

[0045] Example 3: Preparation of compound YYH-1:

[0046] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole was replaced with 2-(chloromethyl)-5-(4-phenyl)-1,3,4-oxadiazole in the above steps, and YYH-1 was obtained by high performance liquid chromatography as a yellow solid with a yield of 66.3%. The structural identification data are as follows:

[0047] 1 H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.01(d,J=7.2Hz,2H),7.87(d,J=8.7Hz,2H),7.64(m,3H),7.12(d,J=8.7Hz,2H),6.82(s,1H), 5.66(s,2H),5.29(s,1H),5.13(t,J=7.0Hz,1H),4.01(s,1H),3.84(s,3H),3.49(m,3H),3.14(m,2H),1.56(s,6H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.4,162.6,161.9,160.7,159.9,157.8,153.4,135.1,132.8,131.9,131.0,130.0(2C),127.1(2 C),123.4,122.7(2C),114.6(2C),108.2(2C),106.0,102.5,96.9,71.6,71.2,70.8,70.6,60.9,56.0,25.8,21.8,18.0(2C).HRMS(ESI + ):[M+H] + calcd for C 36 H 37 N2O 11 ,673.2391; observed 673.2377.

[0048] Example 4: Preparation of compound YYH-2:

[0049] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-oxadiazole, and YYH-2 was obtained by high performance liquid chromatography as a yellow solid with a yield of 81.5%. The structural identification data are as follows:

[0050] 1 H-NMR (600MHz, DMSO-d6): δ12.66(s,1H),8.07(m,2H),7.87(d,J=8.8Hz,2H),7.46(t,J=8.8Hz,2H),7.11(d,J=8.8Hz,2H),6.81(s,1H),5.65(s ,2H),5.28(s,1H),5.12(t,J=7.0Hz,1H),4.01(s,1H),3.84(s,3H),3.4 9(m,3H),3.13(m,2H),1.55(s,3H),1.54(s,3H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.6,164.6,164.0,162.6,161.9,160.7,157.8,153.4,153.1,131.9,131.0,129.9(2C),129.8(2C), 122.7,122.1,117.4,117.2,114.6,108.2(2C),106.0,102.5,97.0,71.6,71.2,70.8,70.6,60.9,56.0,25.8,21.8,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 36 H 36 FN2O 11 ,691.2297; observed 691.2291.

[0051] Example 5: Preparation of compound YYH-3:

[0052] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole was replaced with 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole in the above steps, and YYH-3 was obtained by high performance liquid chromatography as a yellow solid with a yield of 78.4%. The structural identification data are as follows:

[0053] 1H-NMR (600MHz, DMSO-d6): δ8.02(d,J=8.4Hz,2H),7.87(d,J=8.6Hz,2H),7.69(d,J=8.4Hz,2H),7.11(d,J=8.6Hz,2H),6.78(s,1H),5. 63(s,2H),5.31(s,1H),5.12(t,J=6.9Hz,1H),4.05(s,1H),3.85(s,3H),3.50(m,3H),3.16(m,2H),1.55(s,6H),0.80(d,J=5.7Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,164.6,162.8,161.9,160.7,159.9,157.9,153.4,151.9,139.7,137.6,135.1,131.9,131.0,130.2,128.9 ,128.5,125.4,122.7,122.3,122.0,114.6,108.3,106.0,102.5,96.9,71.6,71.2,70.8,70.5,60.9,55.9,34.9,30.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 36 H 36 ClN2O 11 ,708.2080;observed708.2070.

[0054] Example 6: Preparation of compound YYH-5:

[0055] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole was replaced with 2-(chloromethyl)-5-(4-nitrophenyl)-1,3,4-oxadiazole in the above steps. The resulting product, YYH-5, was purified by high-performance liquid chromatography (HPLC) to obtain a yellow solid with a yield of 76.7%. The structural identification data are as follows:

[0056] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.45(d,J=8.8Hz,2H),8.28(d,J=8.8Hz,2H),7.87(d,J=8.7Hz,2H),7.12(d,J=8.7Hz,2H),6.8 4(s,1H),5.71(s,2H),5.28(s,1H),5.14(t,J=7.1Hz,1H),4.00(s,1H),3.85(s,3H),3.50(m,3H),3.14(s,2H),1.57(s,6H),0.79(s,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,164.0,163.6,161.9,160.7,160.0,157.9,149.9(2C),135.2,132.0,131.0,128.9(2C),128.6(2C), 125.2,122.7,122.1,114.6(2C),108.3(2C),106.0,102.5,97.0,71.6,71.2,70.8,70.6,61.0,56.0,25.9,21.8,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 36 H 36 N3O 13 ,718.2242; observed718.2235.

[0057] Example 7: Preparation of compound YYH-6:

[0058] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-(3-nitrophenyl)-1,3,4-oxadiazole, and YYH-6 was obtained by high performance liquid chromatography as a yellow solid with a yield of 83.6%. The structural identification data are as follows:

[0059] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.68(s,1H),8.50(d,J=10.5Hz,1H),8.46(d,J=7.7Hz,1 H),7.94(t,J=8.0Hz,1H),7.89(d,J=8.7Hz,2H),7.13(d,J=8.7Hz,2H),6.84(s,1H),5.72(s,2H), 5.27(s,1H),5.15(m,1H),5.00(d,J=4.4Hz,1H),4.74(d,J=4.4Hz,1H),4.68(d,J=5.6Hz,1H),3.9 9(s,1H),3.85(s,3H),3.47(m,3H),3.14(m,2H),1.59(s,3H),1.56(s,3H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,163.9,163.8,161.9,160.7,160.0,157.9,153.4,148.8,139.7,135.1,133.2,132.0,132.0,131.0,127.2 ,125.4,124.9,122.7,122.1,121.7,114.6,108.3,106.0,102.5,97.0,71.6,71.2,70.6,70.5,61.0,56.0,30.9,25.8,18.0,17.9.HRMS(ESI + ):[M+H] + calcd forC 36 H 36 N3O 13 ,718.2242; observed 718.2235.

[0060] Example 8: Preparation of compound YYH-7:

[0061] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(4-nitrophenyl)vinyl]-1,3,4-oxadiazole, and YYH-7 was obtained by high performance liquid chromatography as a yellow solid with a yield of 61.2%. The structural identification data are as follows:

[0062] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.28(d,J=8.8Hz,2H),8.09(d,J=8.8H z,2H),7.88(d,J=8.1Hz,2H),7.79(d,J=16.5Hz,1H),7.67(d,J=8.5Hz,1H),7.1 3(d,J=8.5Hz,2H),6.81(s,1H),5.64(s,2H),5.28(s,1H),5.13(s,1H),4.00(s, 1H),3.85(s,3H),3.50(m,3H),3.16(m,2H),1.57(s,6H),0.80(d,J=5.6Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.9,164.9,162.3,161.9,160.6,153.4,15 1.9,148.3,141.4,137.5,131.0,130.9,129.5(2C),125.4,124.5(2C),1 23.9,123.6,122.7,122.1,115.6,114.6,114.5,108.2,106.0,102.6,96 .9,71.6,71.2,70.8,70.6,60.9,56.0,30.9,25.9,21.8,18.0.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 N3O 13 ,744.2399; observed 744.2397.

[0063] Example 9: Preparation of compound YYH-8:

[0064] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-nitrophenyl)vinyl]-1,3,4-oxadiazole, and YYH-8 was obtained by high performance liquid chromatography as a yellow solid with a yield of 63.3%. The structural identification data are as follows:

[0065] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.65(s,1H),8.29(d,J=7.8Hz,1H),8.24(d,J=8.2Hz,1H),7.88(d,J=8.9Hz,2H),7.82(d,J=16.5Hz,1H ),7.73(t,J=7.8Hz,1H),7.68(d,J=16.5Hz,1H),7.13(d,J=8.9Hz,2H),6 .81(s,1H),5.64(s,2H),5.27(s,1H),5.13(m,1H),5.00(s,1H),4.75(br s,2H),3.99(s,1H),3.84(s,3H),3.45(m,3H),3.10(m,2H),1.57(s,6H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.0,162.2,161.9,160.7,160.0,157 .9,153.4,148.9,139.7,137.7,136.8,135.1,134.3,132.0,131.0,130.9 ,125.4,124.8,123.1,122.7,122.1,114.1,113.2,108.2,106.0,102.5,9 6.9,71.6,71.2,70.8,70.6,61.0,56.0,30.9,25.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 N3O 13 ,744.2399; observed 744.2396.

[0066] Example 10: Preparation of compound YYH-9:

[0067] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(4-trifluoromethylphenyl)vinyl]-1,3,4-oxadiazole, and YYH-9 was obtained by high performance liquid chromatography as a yellow solid with a yield of 52.6%. The structural identification data are as follows:

[0068] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.02(d,J=8.4Hz,2H),7.88(d,J=8.8H z,2H),7.80(d,J=8.4Hz,2H),7.73(d,J=16.5Hz,1H),7.59(d,J=16.5Hz,1H),7. 13(d,J=8.8Hz,2H),6.81(s,1H),5.63(s,2H),5.28(s,1H),5.13(s,1H),4.00(s ,1H),3.85(s,3H),3.51(m,3H),3.16(m,2H),1.57(s,6H),0.80(d,J=5.9Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.0,162.2,161.9,160.7,160.0,157. 8,153.4,138.9,138.2,135.1,131.9,131.0,130.5,130.3,130.1,129.1(2 C),126.3,125.4,123.6,122.7,122.1,114.6,113.1,108.2,106.0,102.6, 96.9,71.6,71.2,70.8,70.6,60.9,56.0,30.9,25.9,21.8,18.0.HRMS(ESI + ):[M+H] + calcd for C 39 H 38 F3N2O 11 ,767.2422; observed 767.2415.

[0069] Example 11: Preparation of compound YYH-10:

[0070] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-trifluoromethylphenyl)vinyl]-1,3,4-oxadiazole, and YYH-10 was obtained by high performance liquid chromatography as a yellow solid with a yield of 57.3%. The structural identification data are as follows:

[0071] 1H-NMR (600MHz, DMSO-d6): δ12.69(s,1H),8.20(s,1H),8.12(d,J=7.8Hz,1H),7.88(d,J=8.9Hz,2H),7.76(m,2H),7.69(t, J=7.8Hz,1H),7.63(d,J=16.3Hz,1H),7.13(d,J=8.9Hz,2H),6.80(s,1H),5.63(s,2H),5.28(s,1H),5.13(s,1H),5.06(br s,1H),4.85(br s,2H),4.00(s,1H),3.85(s,3H),3.50(m,3H),3.14(m,2H),1.57(s,6H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.1(2C),162.1,161.1,160.7,157.8 ,153.4,138.3,136.0,135.1,132.1,131.9,131.0,130.5,130.4,130.2,12 6.8,125.4,125.1,123.6,122.7,122.1,114.6(2C),112.4,106.0,102.5, 96.9,71.6,71.2,70.8,70.6,60.9,56.0,30.9,25.9,21.8,18.0.HRMS(ESI + ):[M+H] + calcd for C 39 H 38 F3N2O 11 ,767.2422; observed 767.2413.

[0072] Example 12: Preparation of compound YYH-11:

[0073] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(4-fluorophenyl)vinyl]-1,3,4-oxadiazole, and YYH-11 was obtained by high performance liquid chromatography as a yellow solid with a yield of 61.3%. The structural identification data are as follows:

[0074] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.89(d,J=8.9Hz,2H),7.74(d,J=10.2Hz,1H),7.66(d,J= 16.4Hz,1H;d,J=7.8Hz,1H),7.49(m,2H),7.26(t,J=10.2Hz,1H),7.13(d,J=8.9Hz,2H),6.81(s,1H ),5.63(s,2H),5.28(s,1H),5.12(s,1H),5.00(d,J=4.4Hz,1H),4.47(d,J=4.9Hz,1H),4.68(d,J=5 .6Hz,1H),3.99(s,1H),3.85(s,3H),3.45(m,3H),3.14(m,2H),1.58(s,6H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.1,163.7,162.1,162.0,161.9,160 .7,159.9,157.8,153.4,138.7,137.5,137.4,135.1,131.9,131.4,131.0 ,125.0,122.6,122.0,117.4,117.2,114.6,111.7,108.2,105.9,102.5,9 6.9,71.5,71.1,70.7,70.5,60.9,55.9,25.8,21.7,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 FN2O 11 ,717.2454; observed 717.2445.

[0075] Example 13: Preparation of compound YYH-12:

[0076] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-trifluoromethylphenyl)vinyl]-1,3,4-oxadiazole, and YYH-12 was obtained by high performance liquid chromatography as a yellow solid with a yield of 63.3%. The structural identification data are as follows:

[0077] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.89(d,J=8.9Hz,2H),7.74(d,J=10.2Hz,1H),7.66(d,J= 16.4Hz,1H;d,J=7.8Hz,1H),7.49(m,2H),7.26(t,J=10.2Hz,1H),7.13(d,J=8.9Hz,2H),6.81(s,1H ),5.63(s,2H),5.28(s,1H),5.12(s,1H),5.00(d,J=4.4Hz,1H),4.47(d,J=4.9Hz,1H),4.68(d,J=5 .6Hz,1H),3.99(s,1H),3.85(s,3H),3.45(m,3H),3.14(m,2H),1.58(s,6H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.1,163.7,162.1,162.0,161.9,160 .7,159.9,157.8,153.4,138.7,137.5,137.4,135.1,131.9,131.4,131.0 ,125.0,122.6,122.0,117.4,117.2,114.6,111.7,108.2,105.9,102.5,9 6.9,71.5,71.1,70.7,70.5,60.9,55.9,25.8,21.7,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 FN2O 11 ,717.2454; observed 717.2445.

[0078] Example 14: Preparation of compound YYH-13:

[0079] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(4-chlorophenyl)vinyl]-1,3,4-oxadiazole, and YYH-13 was obtained by high performance liquid chromatography as a yellow solid with a yield of 58.9%. The structural identification data are as follows:

[0080] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.89(d,J=8.8Hz,2H),7.84(d,J=8.4Hz,2H),7.66(d,J=16.4Hz,1H),7.52(d,J=8.4H z,2H),7.46(d,J=16.4Hz,1H),7.14(d,J=8.8Hz,2H),6.81(s,1H),5.63(s,2H),5.28(s,1H),5.31(m,1H),5.01(s,1H),4.76(br s,2H),4.00(s,1H),3.85(s,3H),3.45(m,3H),3.13(m,2H),1.58(s,6H),0.80(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.6,162.1,161.9,160.7,160.0,157.9,153.4,139.7,138.7,135.1(2C),133.9,132.0,131.0,130.2,129. 5(2C),125.4,122.7,122.1,114.6(2C),111.0,108.2,106.0,102.5,96 .9,71.6,71.2,70.8,70.6,60.9,56.0,31.0,25.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 ClN2O 11 ,733.2158; observed733.2160.

[0081] Example 15: Preparation of compound YYH-14:

[0082] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-chlorophenyl)vinyl]-1,3,4-oxadiazole, and YYH-14 was obtained by high performance liquid chromatography as a yellow solid with a yield of 56.6%. The structural identification data are as follows:

[0083] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.94(s,1H),7.89(d,J=8.8Hz,2H),7.75(s,1H),7.64(d,J=16.5Hz,1H),7.52(d,J =16.5Hz,1H),7.47(s,2H),7.13(d,J=8.8Hz,2H),6.80(s,1H),5.63(s,2H),5.27(s,1H),5.12(m,1H),5.00(s,1H),4.75(br s,2H),3.99(s,1H),3.85(s,3H),3.45(m,3H),3.10(m,2H),1.57(s,6H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.1,162.0,161.9,160.7,160.0,157 .9,153.4,139.7,138.4,137.2,135.1,134.3,132.0,131.2,131.0,130.2 ,127.9,127.2,125.4,122.7,122.1,114.6,111.9,108.2,106.0,102.5,9 6.9,71.9,71.2,70.8,70.6,60.9,56.0,30.9,25.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 ClN2O 11 ,733.2158; observed733.2152.

[0084] Example 16: Preparation of compound YYH-15:

[0085] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(4-bromophenyl)vinyl]-1,3,4-oxadiazole, and YYH-15 was obtained by high performance liquid chromatography as a yellow solid with a yield of 55.4%. The structural identification data are as follows:

[0086] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.88(d,J=8.9Hz,2H),7.75(d,J=8.5Hz,2H) ,7.64(m,3H),7.45(d,J=16.4Hz,1H),7.13(d,J=8.9Hz,2H),6.80(s,1H),5.61(s,2H), 5.28(s,1H),5.12(s,1H),4.99(d,J=4.4Hz,1H),4.74(d,J=4.4Hz,1H),4.67(s,1H),4 .00(s,1H),3.13(s,3H),3.48(m,3H),3.15(m,2H),1.57(s,6H),0.80(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.2,161.9,160.7,159.9,157.9,153.4,138.7,135.1,134.2,132.4(2C),131.9,131.0(2C),130.3(2C),1 25.4,124.0,122.7,122.1,114.6(2C),111.1,108.2,105.9,102.5,96. 9,71.6,71.2,70.8,70.6,60.9,56.0,30.9,25.9,21.8,18.0.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 BrN2O 11 ,777.1653; observed 767.1647.

[0087] Example 17: Preparation of compound YYH-16:

[0088] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-bromophenyl)vinyl]-1,3,4-oxadiazole, and YYH-16 was obtained by high performance liquid chromatography as a yellow solid with a yield of 57.3%. The structural identification data are as follows:

[0089] 1H-NMR (600MHz, DMSO-d6): δ12.65(s,1H),8.06(s,1H),7.89(d,J=8.9Hz,2H),7.80(d,J=4.4Hz,2H ),7.61(m,2H),7.52(d,J=16.5Hz,1H),7.40(t,J=7.8Hz,1H),7.13(d,J=8.9Hz,2H),6.81(s,1H), 5.63(s,2H),5.27(s,1H),5.12(m,1H),5.00(d,J=4.4Hz,1H),4.74(d,J=4.4Hz,1H),4.68(d,J=5. 7Hz,1H),4.00(s,1H),3.85(s,3H),3.46(m,3H),3.14(m,2H),1.58(s,6H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.1,162.0,161.9,160.7,159.9,157 .9,153.4,139.7,138.4,137.4,135.1,133.1,132.0,131.5,131.0,130.8 ,127.8,125.4,122.9,122.7,122.1,114.1,111.9,108.2,106.0,102.5,9 6.9,71.6,71.2,70.8,70.6,60.9,56.0,30.9,25.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 38 H 38 BrN2O 11 ,777.1653; observed 767.1649.

[0090] Example 18: Preparation of compound YYH-17:

[0091] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(4-methoxyphenyl)vinyl]-1,3,4-oxadiazole, and YYH-17 was obtained by high performance liquid chromatography as a yellow solid with a yield of 51.2%. The structural identification data are as follows:

[0092] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.88(d,J=8.8Hz,2H),7.74(d,J=8.7Hz,2H),7.58(d,J=16.4Hz,1H),7.23(d,J=16.4 Hz,1H),7.13(d,J=8.8Hz,2H),7.00(d,J=8.7Hz,2H),6.78(s,1H),5.59(s,2H),5.28(s,1H),5.13(s,1H),5.04(s,1H),4.28(br s,2H),4.00(s,1H),3.85(s,3H),3.80(s,3H),3.49(m,3H),3.16(m,2H),1.58(s,6H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.7(2C),161.9,161.5,161.4(2C),160.8,153.5,139.8,135.2,131.9,131.0(2C),130.1(2C),127.6,12 5.4,122.7,122.1,114.9(2C),114.6(2C),107.6(2C),102.5,97.0,71 .6,71.2,70.8,70.6,60.9,56.0,55.8,25.9,21.8,18.0(2C).HRMS(ESI + ):[M+H] + calcd for C 39 H 41 N2O 12 ,729.2654; observed729.2646.

[0093] Example 19: Preparation of compound YYH-18:

[0094] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-methoxyphenyl)vinyl]-1,3,4-oxadiazole, and YYH-18 was obtained by high performance liquid chromatography as a yellow solid with a yield of 52.2%. The structural identification data are as follows:

[0095] 1H-NMR (600MHz, DMSO-d6): δ12.68(s,1H),8.21(s,1H),8.13(d,J=7.7Hz,1H),7.89(d,J=8. 7Hz,2H),7.77(m,2H),7.68(t,J=7.7Hz,1H),7.63(d,J=16.5Hz,1H),7.13(d,J=8.7Hz,2H) ,6.81(s,1H),5.64(s,2H),5.27(s,1H),5.13(s,1H),5.00(s,1H),4.74(s,1H),4.68(s,1H ),3.99(s,1H),3.85(s,3H),5.00(m,3H),3.13(m,2H),1.57(s,6H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.1,162.0,161.9,160.7,159.9,15 7.8,153.4,138.3,136.1,135.1,132.1,131.9,131.0,130.4,126.7,125. 1,122.6,122.0,114.5(2C),112.3(2C),108.2(2C),105.9,102.5,96.9, 71.5,71.1,70.7,70.5,60.9,55.9(2C),25.8,21.7,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 39 H 41 N2O 12 ,729.2654; observed 729.2642.

[0096] Example 20: Preparation of compound YYH-19:

[0097] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-(dimethylamino)phenyl)vinyl]-1,3,4-oxadiazole, and YYH-19 was obtained by high performance liquid chromatography as a yellow solid with a yield of 65.6%. The structural identification data are as follows:

[0098] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.89(d,J=8.9Hz,2H),7.58(d,J=8.9Hz,2H),7. 48(d,J=16.3Hz,1H),7.13(d,J=8.9Hz,2H),7.03(d,J=16.3Hz,1H),6.80(s,1H),6.73(d,J =8.9Hz,2H),5.58(s,2H),5.28(s,1H),5.13(m,1H),5.01(s,1H),4.76(s,2H),4.00(s,1H ),3.85(s,3H),3.44(m,3H),3.13(m,2H),2.98(s,6H),1.57(s,6H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,166.2,161.9,161.1,160.8,160.0,157 .8,153.4,1520.0,140.5,135.1,132.4,131.9,131.0,129.9(2C),122.7(2 C),122.3(2C),122.0(2C),114.6(2C),112.3,111.6,105.9,104.0,102.5, 96.9,71.6,71.2,70.8,70.6,60.9,56.0,25.9,21.8,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 40 H 44 N3O 11 ,724.2970; observed 724.2964.

[0099] Example 21: Preparation of compound YYH-20:

[0100] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-phenylvinyl]-1,3,4-oxadiazole, and YYH-20 was obtained by high performance liquid chromatography as a yellow solid with a yield of 48.5%. The structural identification data are as follows:

[0101] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.87(d,J=8.9Hz,2H),7.77(d,J=6.7Hz,2H),7.61(d, J=16.5Hz,1H),7.43(d,J=7.4Hz,2H),7.37(d,J=16.5Hz,1H),7.27(t,J=8.8Hz,1H),7.12(d,J= 8.9Hz,2H),6.81(s,1H),5.62(s,2H),5.27(s,1H),5.13(m,1H),5.00(s,1H),4.74(s,1H),4.68 (s,1H),4.00(s,1H),3.85(s,3H),3.50(m,3H),3.14(m,2H),1.58(s,6H),0.79(d,J=6.1Hz,3H) 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.4,161.9,161.8,160.7,159.9,157.9,153.4,140.1,135.1,134.8,132.0,130.9,130.7,129.5(2C),128 .4(2C),122.6(2C),122.0,114.6(2C),110.1,108.2,105.9,102.5,96. 9,71.6,71.2,70.8,70.5,60.9,56.0,30.9,25.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 38 H 39 N2O 11 ,699.2548; observed 699.2534.

[0102] Example 22: Preparation of compound YYH-21:

[0103] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-(dioxomethylene)phenyl)vinyl]-1,3,4-oxadiazole, and YYH-21 was obtained by high performance liquid chromatography as a yellow solid with a yield of 56.4%. The structural identification data are as follows:

[0104] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),7.89(d,J=8.9Hz,2H),7.55(d,J=16.3Hz,1H),7.52(s,1H),7. 26(d,J=16.3Hz,1H),7.23(d,J=8.1Hz,1H),7.13(d,J=8.9Hz,2H),6.98(d,J=8.1Hz,1H),6.80(s,1H),6 .09(s,2H),5.60(s,2H),5.27(s,1H),5.12(m,1H),5.00(d,J=4.5Hz,1H),4.74(d,J=4.5Hz,1H),4.68(d ,J=5.6Hz,1H),3.99(s,1H),3.85(s,3H),3.45(m,3H),3.14(m,2H),1.57(s,6H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,165.6,161.9,161.6,160.8,159.9,157. 9,153.4,149.6,148.6,139.9,139.7,135.1,132.0,131.0,129.4,125.4,12 5.0,122.7,122.0,114.6,109.0,108.2,108.1,106.7,105.9,102.6,102.1 ,96.9,71.6,71.2,70.8,70.6,60.9,56.0,30.9,25.9,18.0,17.9.HRMS(ESI + ):[M+H] + calcd for C 39 H 39 N2O 13 ,743.2446; observed743.2445.

[0105] Example 23: Preparation of compound YYH-22:

[0106] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 5-(3-{4-[bis(2-chloroethyl)amino]phenyl}propyl)-2-(chloromethyl)-1,3,4-oxadiazole, and YYH-22 was obtained by high performance liquid chromatography as a yellow solid with a yield of 44.7%. The structural identification data are as follows:

[0107] 1H-NMR (600MHz, DMSO-d6): δ12.66(s,1H),7.89(d,J=8.8Hz,2H),7.14(d,J=8.8Hz,2H),7.02(d,J=8.6Hz,2H),6. 77(s,1H),6.66(d,J=8.6Hz,2H),5.56(s,2H),5.28(d,J=1.3Hz,2H),5.12(m,1H),5.00(d,J=4.5Hz,1H),4.74(d ,J=4.5Hz,1H),4.68(d,J=5.8Hz,1H),4.00(m,1H),3.86(s,3H),3.69(t,J=4.1Hz,8H),3.49(m,1H),3.44(m,2H) ,3.15(m,2H),2.87(t,J=7.3Hz,2H),2.19(s,1H),1.94(m,2H),1.57(s,5H),1.36(s,4H),0.80(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.3,167.6,161.9,161.4,160.2,159.4,157.4, 152.9,144.6,134.6,131.5,130.5,129.3(2C),129.0(2C),124.9,122.2,114. 1(2C),111.9(2C),107.7,105.4,102.1,96.4,71.1,70.7,70.3,70.1,67.0,5 5.5,52.2(2C),41.1,(2C),34.4,33.0,27.9,25.4,21.3,17.5,17.5.HRMS(ESI + ):[M+H] + calcd forC 43 H 51 Cl2N3O 11 ,853.2864; observed 853.2853.

[0108] Example 24: Preparation of compound YYH-24:

[0109] According to the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above steps was replaced with 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-thiadiazole, and YYH-24 was obtained by high performance liquid chromatography as a yellow solid with a yield of 36.3%. The structural identification data are as follows:

[0110] 1H-NMR (600MHz, DMSO-d6): δ12.66(s,1H),8.03(d,J=8.4Hz,2H),7.88(d,J=8.7Hz,2 H),7.64(d,J=8.4Hz,2H),7.13(d,J=8.7Hz,2H),6.80(s,1H),5.81(s,2H),5.27(s, 1H),5.14(t,J=6.6Hz,1H),5.00(s,1H),4.74(s,1H),4.68(s,1H),4.00(s,1H),3.8 5(s,3H),3.48(m,3H),3.09(m,2H),1.62(s,3H),1.60(s,3H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.7,168.6,166.5,161.9,160.7,159.9,157.8,153.4,151.9,139.6,136.7,135.12,132.0,131.0,130.0,129. 9,128.6,128.5,125.3,122.6,122.2,114.5,108.1,105.8,102.5,97. 0,71.5,71.8,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS(ESI + ):[M+H] + calcd for C 36 H 36 ClN2O 10 S,723.1773; observed 723.1771.

[0111] Example 25: Preparation of compound YYH-25:

[0112] According to the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above steps was replaced with 2-(chloromethyl)-5-(4-bromophenyl)-1,3,4-thiadiazole, and YYH-25, a yellow solid, was obtained by high performance liquid chromatography with a yield of 34.3%. The structural identification data are as follows:

[0113] 1H-NMR (600MHz, DMSO-d6): δ12.66(s,1H),7.94(d,J=8.3Hz,2H),7.87(d,J=8.8Hz,2 H),7.77(d,J=8.3Hz,2H),7.13(d,J=8.8Hz,2H),6.80(s,1H),5.80(s,2H),5.28(s, 1H),5.13(t,J=6.8Hz,1H),5.00(s,1H),4.75(s,1H),4.69(s,1H),4.00(s,1H),3.8 5(s,3H),3.48(m,3H),3.13(m,2H),1.61(s,3H),1.59(s,3H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.7,168.7,166.5,161.9,160.6,159.9,157.8,153.4,151.9,139.6,135.1,132.9,132.0,131.0,130.0,128.9 ,128.5,125.5,125.3,122.6,122.2,114.5,108.1,105.8,102.5,96.9,71.5,71.1,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS (ESI + ):[M+H] + calcd for C 36 H 36 BrN2O 10 S,767.1268; observed 767.1255.

[0114] Example 26: Preparation of compound YYH-26:

[0115] Following the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above steps was replaced with 2-(chloromethyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole, and YYH-26 was obtained by high performance liquid chromatography as a yellow solid with a yield of 38.2%. The structural identification data are as follows:

[0116] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.23(d,J=7.6Hz,2H),7.93(d,J=7. 6Hz,2H),7.87(d,J=8.2Hz,2H),7.13(d,J=8.2Hz,2H),6.79(s,1H),5.84(s,2H ),5.28(s,1H),5.15(s,1H),5.03(s,1H),4.78(s,2H),4.00(s,1H),3.85(s,3 H),3.48(m,3H),3.14(m,2H),1.62(s,3H),1.60(s,3H),0.79(d,J=5.3Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.7,168.3,167.3,161.8,160.6,157.7,153.4,151.9,139.6,135.1,133.4,131.9,131.7,131.5,129.0,126.9,12 6.8,125.3,125.1,123.3,122.6,122.2,114.5,108.0,106.0,102.5,97 .0,71.5,71.1,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS(ESI + ):[M+H] + calcd for C 37 H 36 F3N2O 10 S,757.2037; observed757.2026.

[0117] Example 27: Preparation of compound YYH-27:

[0118] According to the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above steps was replaced with 2-(chloromethyl)-5-(3-fluorophenyl)-1,3,4-thiadiazole, and YYH-27 was obtained by high performance liquid chromatography as a yellow solid with a yield of 36.6%. The structural identification data are as follows:

[0119] 1H-NMR (600MHz, DMSO-d6): δ12.66 (s, 1H), 8.06 (m, 2H), 7.87 (d, J = 8.4Hz, 2H), 7 .41(t,J=8.5Hz,2H),7.12(d,J=8.4Hz,2H),6.80(s,1H),5.80(s,2H),5.28(s, 1H),5.14(s,1H),5.00(s,1H),4.74(s,1H),4.68(s,1H),4.00(s,1H),3.85(s, 3H), 3.48 (m, 3H), 3.14 (m, 2H), 1.61 (s, 3H), 1.60 (s, 3H), 0.79 (d, J = 5.7Hz, 3H). 13 C-NMR (150MHz, DMSO-d6): δ178.8,168.6,166.2,161.9,160.7,159.9,157.8,153.4,151.9,139.6,135.1,132.0,131.0,130.7,130.6,128.5 ,126.4,125.3,122.6,122.2,117.1,114.5,108.1,105.8,102.5,96.9,71.5,71.1,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS(ESI + ):[M+H] + calcd for C 36 H 36 FN2O 11 S,707.2069; observed707.2059.

[0120] Example 28: Preparation of compound YYH-28:

[0121] According to the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above steps was replaced with 2-(chloromethyl)-5-(3-chlorophenyl)-1,3,4-thiadiazole, and YYH-28 was obtained by high performance liquid chromatography as a yellow solid with a yield of 35.5%. The structural identification data are as follows:

[0122] 1H-NMR (600MHz, DMSO-d6): δ12.66(s,1H),8.03(d,J=8.4Hz,2H),7.88(d,J=8.7Hz,2 H),7.64(d,J=8.4Hz,2H),7.13(d,J=8.7Hz,2H),6.80(s,1H),5.81(s,2H),5.27(s, 1H),5.14(t,J=6.6Hz,1H),5.00(s,1H),4.74(s,1H),4.68(s,1H),4.00(s,1H),3.8 5(s,3H),3.48(m,3H),3.09(m,2H),1.62(s,3H),1.60(s,3H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.7,168.6,166.5,161.9,160.7,159.9,157.8,153.4,151.9,139.6,136.7,135.12,132.0,131.0,130.0,129. 9,128.6,128.5,125.3,122.6,122.2,114.5,108.1,105.8,102.5,97. 0,71.5,71.8,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS(ESI + ):[M+H] + calcd for C 36 H 36 ClN2O 10 S,723.1773; observed 723.1771.

[0123] Example 29: Preparation of compound YYH-29:

[0124] According to the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above steps was replaced with 2-(chloromethyl)-5-(3-bromophenyl)-1,3,4-thiadiazole, and YYH-29 was obtained by high performance liquid chromatography as a yellow solid with a yield of 31.9%. The structural identification data are as follows:

[0125] 1H-NMR (600MHz, DMSO-d6): δ12.66(s,1H),7.94(d,J=8.3Hz,2H),7.87(d,J=8.8Hz,2 H),7.77(d,J=8.3Hz,2H),7.13(d,J=8.8Hz,2H),6.80(s,1H),5.80(s,2H),5.28(s, 1H),5.13(t,J=6.8Hz,1H),5.00(s,1H),4.75(s,1H),4.69(s,1H),4.00(s,1H),3.8 5(s,3H),3.48(m,3H),3.13(m,2H),1.61(s,3H),1.59(s,3H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.7,168.7,166.5,161.9,160.6,159.9,157.8,153.4,151.9,139.6,135.1,132.9,132.0,131.0,130.0,128.9 ,128.5,125.5,125.3,122.6,122.2,114.5,108.1,105.8,102.5,96.9,71.5,71.1,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS (ESI + ):[M+H] + calcd for C 36 H 36 BrN2O 10 S,767.1268; observed 767.1255.

[0126] Example 30: Preparation of compound YYH-30:

[0127] According to the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above steps was replaced with 2-(chloromethyl)-5-(3-trifluoromethylphenyl)-1,3,4-thiadiazole, and YYH-30 was obtained by high performance liquid chromatography as a yellow solid with a yield of 38.1%. The structural identification data are as follows:

[0128] 1H-NMR (600MHz, DMSO-d6): δ12.67(s,1H),8.23(d,J=7.6Hz,2H),7.93(d,J=7. 6Hz,2H),7.87(d,J=8.2Hz,2H),7.13(d,J=8.2Hz,2H),6.79(s,1H),5.84(s,2H ),5.28(s,1H),5.15(s,1H),5.03(s,1H),4.78(s,2H),4.00(s,1H),3.85(s,3 H),3.48(m,3H),3.14(m,2H),1.62(s,3H),1.60(s,3H),0.79(d,J=5.3Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.7,168.3,167.3,161.8,160.6,157.7,153.4,151.9,139.6,135.1,133.4,131.9,131.7,131.5,129.0,126.9,12 6.8,125.3,125.1,123.3,122.6,122.2,114.5,108.0,106.0,102.5,97 .0,71.5,71.1,70.7,70.5,65.3,55.9,30.8,25.9,18.2,17.9.HRMS(ESI + ):[M+H] + calcd for C 37 H 36 F3N2O 10 S,757.2037; observed757.2026.

[0129] Example 31: Preparation of compound YYH-31:

[0130] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-[(1E)-2-(3-methoxyphenyl)vinyl]-1,3,4-oxadiazole, and YYH-31 was obtained by high performance liquid chromatography as a yellow solid with a yield of 46.1%. The structural identification data are as follows:

[0131] 1H-NMR (600MHz, DMSO-d6): δ12.65(s,1H),7.88(d,J=8.8Hz,2H),7.12(d,J=8.8Hz,2H),6.76(s,1H),6.53(m,1 H),5.57(s,2H),5.27(s,1H),5.12(t,J=7.1Hz,1H),5.03(s,1H),4.80(s,2H),3.99(s,1H),3.85(s,3H),3.49( dt,J=9.2,4.3Hz,1H),3.44(dd,J=14.3,7.0Hz,1H),3.40–3.35(m,1H),3.14(t,J=9.3Hz,1H),3.09(q,J=7.5H z,1H),2.27(p,J=7.4Hz,2H),2.05(s,3H),1.58(d,J=3.1Hz,6H),1.03(t,J=7.5Hz,3H),0.79(d,J=6.0Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.79,166.84,161.90,161.82,160.71,159.95 ,157.83,153.37,140.03,135.11,131.88,130.99(2C),122.68,122.03,119 .90,114.59(2C),108.13,105.92,102.53,96.90,71.59,71.19,70.79,70.5 5,60.89,55.97,25.88,21.75,21.64,17.96,17.93,13.54,12.90.HRMS(ESI + ):[M+H] + calcd forC 35 H 41 N2O 11 ,665.2705; observed 665.2681.

[0132] Example 32: Preparation of compound YYH-32:

[0133] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-n-octyl-1,3,4-oxadiazole, and YYH-32 was obtained by high performance liquid chromatography as a yellow solid with a yield of 86.7%. The structural identification data are as follows:

[0134] 1H NMR (600MHz, DMSO-d6): δ12.62(s,1H),7.87(d,J=8.8Hz,2H),7.13(d,J=8.8Hz,2H),6.75(s,1H),5.56(s,2 H),5.27(s,1H),5.11(t,J=7.2Hz,1H),5.01(s,1H),4.76(s,2H),3.99(s,1H),3.85(s,3H),3.51–3.46(m,1H ),3.43(dd,J=14.9,7.2Hz,1H),3.39–3.35(m,1H),3.14(t,J=9.3Hz,1H),3.11–3.04(m,1H),2.87(t,J=7.3 Hz,2H),1.67(p,J=7.2Hz,2H),1.58(s,6H),1.30–1.13(m,8H),0.81(t,J=7.1Hz,3H),0.78(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.31,167.76,161.84,161.42,160.19,159.41,157 .35,152.89,134.62,131.40,130.49(2C),122.18,121.55,114.10(2C),107.69, 105.41,102.04,96.42,71.09,70.69,70.29,70.05,69.76,60.29,55.49,31.00, 28.14,28.08,25.80,25.40,24.37,21.96,21.23,17.50,17.43,13.85.HRMS(ESI + ):[M+H] + calcdfor C 35 H 41 N2O 11 ,695.3174; observed695.3176.

[0135] Example 33: Preparation of compound YYH-33:

[0136] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole was replaced with 2-(chloromethyl)-5-cyclopropyl-1,3,4-oxadiazole in the above steps. The resulting product, YYH-33, was purified by high-performance liquid chromatography (HPLC) to obtain a yellow solid with a yield of 47.3%. The structural identification data are as follows:

[0137] 1H NMR (600MHz, DMSO-d6) δ12.65(s,1H),7.88(d,J=8.9Hz,2H),7.13(d,J=8.9Hz,2H),6.75(s,1H),5.50(s,2H),5.27(s,1H ),5.11(t,J=7.2Hz,1H),4.99(d,J=4.4Hz,1H),4.73(d,J=5.0Hz,1H),4.67(d,J=5.8Hz,1H),3.99(s,1H),3.85(s,3H),3. 51–3.46(m,1H),3.43(dd,J=14.5,7.1Hz,1H),3.36(dd,J=14.5,7.2Hz,1H),3.14(td,J=9.3,5.0Hz,1H),3.11–3.05(m,1H ),2.28(ddd,J=13.3,8.4,4.9Hz,1H),1.59(d,J=9.9Hz,6H),1.20–1.13(m,2H),1.03–0.97(m,2H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.81,169.60,161.91,161.60,160.71,159.89,157.88,153.37,135.10,131.94,131.00(2C),122.68,122.00,1 14.60(2C),108.16,105.89,102.52,96.88,71.56,71.18,70.77,70.5 4,60.77,55.98,25.89,21.74,17.95,17.93,8.63(2C),6.21.HRMS(ESI + ):[M+H] + calcd for C 33 H 37 N2O 11 ,637.2392; observed637.2368.

[0138] Example 34: Preparation of compound YYH-34:

[0139] Following the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above steps was replaced with 2-(chloromethyl)-5-cyclohexyl-1,3,4-oxadiazole, and YYH-34 was obtained by high performance liquid chromatography as a yellow solid with a yield of 62.2%. The structural identification data are as follows:

[0140] 1H NMR (600MHz, DMSO-d6) δ12.66(s,1H),7.88(d,J=8.9Hz,2H),7.13(d,J=8.9Hz,2H),6.76(s,1H),5.55(s,2H),5.29–5.26(m,1H),5.12 (t,J=7.2Hz,1H),4.99(d,J=4.4Hz,1H),4.73(d,J=5.0Hz,1H),4.67(d,J=5.8Hz,1H),3.99(t,J=4.6Hz,1H),3.85(s,3H),3.51–3.41( m,2H),3.37(dd,J=14.5,7.2Hz,1H),3.19–3.04(m,2H),3.00(tt,J=10.9,3.7Hz,1H),2.03–1.95(m,2H),1.72(dt,J=12.9,3.7Hz,2H) ,1.67–1.61(m,1H),1.58(d,J=8.0Hz,6H),1.52(dd,J=17.6,6.5Hz,2H),1.44–1.33(m,2H),1.31–1.21(m,1H),0.79(d,J=6.1Hz,3H). 13 C-NMR (150MHz, DMSO-d6): δ178.81,170.94,162.10,161.91,160.73,159.89 ,157.88,153.37,135.10,131.92,131.00(2C),122.68,122.01,114.60(2C) ,108.17,105.89,102.52,96.91,71.56,71.18,70.77,70.54,60.89,55.98, 34.49,29.98(2C),25.86,25.57,25.03(2C),21.74,17.98,17.93.HRMS(ESI + ):[M+H] + calcd forC 35 H 41 N2O 11 ,679.2861; observed.679.2838.

[0141] Example 35: Screening for anti-hepatocellular carcinoma activity

[0142] To demonstrate the potential therapeutic effects of the oxadiazole derivative and thiadiazole derivative YYH, based on icariin II as the parent nucleus, on liver cancer, the compounds prepared above were subjected to pharmacodynamic screening for liver cancer. The specific screening methods and results for the anti-liver cancer compounds are as follows:

[0143] Compounds were initially screened using the MTT assay, with icaradin, icariin-II (ICA-II), and sorafenib (SORA) as positive controls. Well-growing hepatocellular carcinoma cells or normal hepatocytes were selected, digested with trypsin, dispersed in complete cell culture medium to form a single-cell suspension, and then counted to adjust the cell concentration to 5 × 10⁶ cells / year. 4 Cells were dispersed evenly by gentle pipetting and layered into 96-well plates (100 μL / well) and incubated overnight in a CO2 incubator. The next day, the culture medium was discarded, and the test drug was added at concentrations of 0.78125, 1.5625, 3.125, 6.25, 12.5, and 25 μM, respectively. A blank control group was also included to remove background interference. Each group had three replicates. After incubating the samples in a CO2 incubator for 72 hours, the 96-well plates were removed, and 20 μL / well of pre-prepared MTT solution was added, and the plates were incubated for another 4 hours. The MTT solution was then discarded, and 150 μL of DMSO solution was added to each well to dissolve the sample. The plates were shaken at room temperature for 5 minutes to ensure complete dissolution of any crystals. The absorbance of each well was measured using a microplate reader set to OD 490 nm. The experiment was repeated three times. Cell growth inhibition rate was calculated using the following formula: Inhibition rate (%) = (Control A - Added A) / (Control A - Blank A) × 100%. Inhibition rates at different concentrations were calculated, and dose-response curves were plotted. The concentration at which 50% growth inhibition was achieved was calculated as the IC50. 50 Values. The results showed that most compounds had a considerable degree of inhibitory activity against the proliferation of liver cancer cells, as shown in Table 1.

[0144] Table 1: Inhibitory activity of compound YYH against liver cancer cell lines and normal human cells;

[0145]

[0146]

[0147] In summary, by applying the above pharmacological experiments to screen antitumor drugs for the proliferation of liver cancer cells, it was found that compounds such as YYH-7, 9, 11, 17, and 21 exhibit significantly better inhibitory activity against HepG2 liver cancer cells than the positive control drugs (Acoradine, SORA, and ICA-II), and have potential for further development. Therefore, the diazole heterocyclic derivative of icariin II in this invention has good anti-proliferative activity against liver cancer cells and has the potential to be further developed into a drug for the treatment of liver cancer.

[0148] Although specific embodiments of the present invention have been described in detail, those skilled in the art can make various modifications and substitutions to the details of the technical solutions of the present invention based on all the teachings disclosed, and all such modifications and substitutions are within the scope of protection of the present invention. The full scope of protection of the present invention is given by the appended claims and any equivalents thereof.

Claims

1. An icariin II derivative or a pharmaceutically acceptable salt thereof with anti-hepatocellular carcinoma activity, characterized in that: The icariin II derivative is an icariin II oxadiazole derivative; wherein the icariin II oxadiazole derivative has the chemical structure shown in formula (I) or formula (II): ， In equation (Ⅰ), R is selected from: , , , , , , , , , , , , , or ; In equation (II), R is selected from: , , , , , , , , , , , , , , or .

2. An icariin II derivative or a pharmaceutically acceptable salt thereof with anti-hepatocellular carcinoma activity, characterized in that: The icariin II oxadiazole derivative is selected from the following compounds: 。 3. The method for preparing the icariin II derivative or its pharmaceutically acceptable salt as described in claim 2, characterized in that: Includes the following steps: ， Starting from a2-a22, a31, and a33-a34, intermediates b2-b22, b31, and b33-b34 are obtained through step (i). Then, fragments c2-c22, c31, and c33-c34 are obtained through step (ii). Finally, icariin II is replaced through step (iii) to obtain compounds YYH2-22, YYH31, and YYH33-34.

4. The preparation method according to claim 3, characterized in that: The preparation method includes the following steps: (i) Synthesis of b2-b22, b31, b33-b34: Dissolve a2-a22, a31, a33-a34 in a solvent, add 1.5-3 equivalents of HOBt and 1.5-3 equivalents of EDCI, stir for 0.5-4 hours, add 2-4 equivalents of 80% hydrazine hydrate dropwise, react at room temperature, remove the solvent, extract, wash, dry, filter, and purify; (ii) Synthesis of c2-c22, c31, c33-c34: Dissolve b2-b22, b31, b33-b34 in a solvent, add 1-3 equivalents of trimethoxychloroethane and 3-5 equivalents of acetic acid, react at 110-130℃, and the post-treatment is the same as in step (i). (iii) Synthesis of YYH2-22, YYH31, and YYH33-34: Icariin II and 1-2 equivalents of K2CO3 and 0.5-1 equivalents of KI were dissolved in a solvent, stirred at room temperature, and the corresponding fragments were added. The reaction was carried out at 60°C, and the post-treatment was the same as in step (i).

5. The preparation method according to claim 4, characterized in that: The solvent is selected from anhydrous acetonitrile. N,N -Dimethylaminocarbamate, 1,4-dioxane, toluene, or acetone.

6. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the icariin II derivative of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition according to claim 6, characterized in that: The dosage form of the pharmaceutical composition is an oral dosage form or an injectable dosage form.

8. The pharmaceutical composition according to claim 7, characterized in that: The oral dosage form is a capsule, tablet, granule, oral liquid, sustained-release preparation or controlled-release preparation, and the pharmaceutically acceptable carrier is selected from one or more of diluents, lubricants, binders, disintegrants, stabilizers or solvents.

9. The use of the icariin II derivative of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 6 to 8, in the preparation of a medicament for treating primary liver cancer.

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