Icariside II derivative with anti-hepatoma activity as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to icariside II derivatives with anti-hepatoma activity as well as a preparation method and application of the icariside II derivatives. The invention prepares a series of icariside II (ICA-II) derivatives with higher anti-hepatoma activity, and further provides a pharmaceutical composition containing the derivatives or pharmaceutically acceptable salts thereof, and a preparation method of the derivatives or the pharmaceutically acceptable salts thereof. In addition, the invention also provides application of the derivative or the pharmaceutically acceptable salt or the pharmaceutical composition thereof in preparation of drugs for treating primary liver cancer.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to an icariin II derivative with anti-liver cancer activity and a preparation method and application thereof. Background Art

[0002] Liver cancer is one of the most common tumors in the world and the leading cause of cancer-related deaths. Primary liver cancer (Hepatocellular carcinoma, HCC) is the most common form of cancer, accounting for about 90% of the patient population. The causes of liver cancer include (for example, type B and type C) viral hepatitis, alcohol, obesity, dietary carcinogens, etc. The current treatments for liver cancer mainly include surgical resection and liver transplantation, but both are ineffective. Targeted therapies for liver cancer are limited to sorafenib, lenvatinib, regorafenib, ramucirumab, and cabozantinib, which are beneficial for prolonging patient survival, but they are drug-resistant (European Journal of Medicinal Chemistry, 2021, 224: 113690). Therefore, there is an urgent need to develop alternative treatment strategies.

[0003] Icaritin (ICT), as a main component of the traditional Chinese medicine Epimedium, is an isopentenyl flavonoid compound and has been approved for marketing as a Class I innovative drug (the drug name is Acoradine). However, it has poor water solubility and general anti-liver cancer activity. Based on the previous research on its structure-activity relationship by the inventor's team, it was found that (1) the activity of rhamnose at the C3 position is greatly improved; (2) the isopentenyl group is a key group with anti-liver cancer activity; (3) the introduction of a nitrogen-containing heterocycle at the C7 position can enhance the proliferation inhibition effect. Based on this research, the present invention continues to modify the C7 position in order to further improve the anti-liver cancer cell proliferation activity of icariin and its structural analogs.

[0004] 1,3,4-oxadiazole derivatives have been successfully used as anti-mitotic drugs to treat cancer. Most anti-mitotic drugs target microtubules, which are dynamic elements in the cytoskeleton responsible for the formation of the mitotic spindle and are necessary for chromosome separation during cell division. Oxazole derivatives can inhibit tubulin polymerization and prevent tumor cell mitosis, thereby exerting anti-tumor activity (Bioorganic & Medicinal Chemistry Letters, 2006, 16: 1191-1196). In addition, 1,3,4-oxadiazole and 1,3,4-thiadiazole are also hot fragments in anti-tumor research, so the inventors try to introduce them to improve the anti-proliferation activity of icariin against liver cancer cells. Summary of the invention

[0005] In order to solve the above technical problems, the inventors have prepared a series of icariin II (ICA-II) derivatives with high anti-liver cancer activity, and further provided a pharmaceutical composition containing the derivative or a pharmaceutically acceptable salt thereof, and a method for preparing the derivative or a pharmaceutically acceptable salt thereof. In addition, the invention also provides the use of the derivative or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof in the preparation of a drug for treating primary liver cancer.

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

[0007] In the first aspect, the present invention provides an icariin II derivative or a pharmaceutically acceptable salt thereof having anti-hepatocarcinoma 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] R in the formula (I), (II), (III) or (IV) is selected from:

[0010]

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

[0012]

[0013]

[0014]

[0015]

[0016] In a second aspect, the present invention further provides a method for preparing the icariin II oxadiazole derivative or icariin II thiadiazole derivative having anti-hepatocarcinoma activity as described in the first aspect, comprising the following steps:

[0017]

[0018] Using a1-a22 and a31-a34 as starting materials, step (i) is performed to obtain intermediates b1-b22 and b31-34, and then step (ii) is performed to obtain fragments c1-c22 and c31-34, and finally step (iii) is performed to replace icariin II to obtain compounds YYH1-22 and YYH31-34;

[0019] Using a23-a30 as starting materials, step (i) is used to obtain intermediates b23-b30, and step (iv) is used to obtain intermediates c23-c30. Then, step (v) is used to obtain fragment d23-30. Finally, step (iii) is used to replace icariin II to obtain compound YYH23-30.

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

[0021] (i) Synthesis of b1-b34: a1-a34 is dissolved in a solvent, 1.5-3 equivalents of HOBt and 1.5-3 equivalents of EDCI are added, stirred for 0.5-4 hours, 2-4 equivalents of 80% hydrazine hydrate are added dropwise, reacted at room temperature, the solvent is removed, extracted, washed, dried, filtered, and purified;

[0022] (ii) Synthesis of c1-c22 and c31-34: dissolve b1-b22 and b31-34 in a solvent, add 1-3 equivalents of trimethoxyethyl chloride and 3-5 equivalents of acetic acid, react at 110-130°C, and post-treat in the same manner as step (i);

[0023] (iii) Synthesis of YYH1-34: Dissolve icariin I, 1-2 equivalents of K2CO3, and 0.5-1 equivalents of KI in a solvent, stir at room temperature, add the corresponding fragment, react at 60°C, and post-treat in the same manner as step (i);

[0024] (iv) Synthesis of d23-d30: b23-b30 was dissolved in a solvent, 1-2 equivalents of p-chloroacetic acid, 1.5-3 equivalents of HOBt and 1.5-3 equivalents of EDCI were added, and the reaction was carried out at room temperature. The post-treatment method was the same as step (i);

[0025] (v) Synthesis of e23-e30: Dissolve d23-d30 in a solvent, add 2-3 equivalents of Lawesson's reagent, react at 110-130°C, and post-treat in the same manner as step (i);

[0026] 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 the icariin II oxadiazole derivative or icariin II thiadiazole derivative with anti-hepatocarcinoma activity as described in the first aspect above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0029] The dosage form of the pharmaceutical composition is an oral dosage form or an injection dosage form, wherein the oral dosage form is a capsule, a tablet, a granule, an oral liquid, a sustained-release preparation or a controlled-release preparation.

[0030] Preferably, the pharmaceutically acceptable carrier is selected from one or more of a diluent, a lubricant, a binder, a disintegrant, a stabilizer or a solvent.

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

[0032] In the fourth aspect, the present invention provides the use of the icariin II oxadiazole derivative or icariin II thiadiazole derivative with anti-hepatocarcinoma activity described in the first aspect or a pharmaceutically acceptable salt thereof or the pharmaceutical composition described in the third aspect in the preparation of a drug for treating primary liver cancer.

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

[0034] The present invention provides a series of oxadiazole derivatives or thiadiazole derivatives or pharmaceutically acceptable salts thereof with icariin II as the mother nucleus. The present invention provides a lead compound for preparing a drug for treating primary liver cancer. DETAILED DESCRIPTION

[0035] The present invention is further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention. If specific techniques or conditions are not specified in the embodiments, they are carried out according to the techniques or conditions described in the literature in this field, or according to the product instructions. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased through regular channels.

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

[0037]

[0038] Take a 25mL reaction bottle, put ICA-II (50mg, 0.097mmoL), anhydrous potassium carbonate (27mg, 0.194mmoL), potassium iodide (8mg, 0.0485mmol) and 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole (44mg, 0.194mmol) in it, add 3mL dry acetone to dissolve, and stir at 60℃ for 16h. TLC monitors the reaction completion, adds 2mL water to quench the reaction, adds 2mL ethyl acetate to extract three times, washes the organic phase with saturated sodium chloride solution, and removes the solvent under reduced pressure after drying with anhydrous sodium sulfate. Purify by RP-HPLC to obtain YYH-4 with a yield of 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] + Calculate for C 37 H 39 N2O 12,703.2497; observed 703.2492.

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

[0042]

[0043] Take a 25mL reaction bottle, put ICA-II (50mg, 0.097mmoL), anhydrous potassium carbonate (27mg, 0.194mmoL), potassium iodide (8mg, 0.0485mmol) and 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-thiadiazole fragment (41mg, 0.194mmol) in it, add 3mL dry acetone to dissolve, and stir at 60℃ for 16h. TLC monitors the reaction completion, adds 2mL water to quench the reaction, adds 2mL ethyl acetate to extract three times, washes the organic phase with saturated sodium chloride solution, and removes the solvent under reduced pressure after drying with anhydrous sodium sulfate. Purify by RP-HPLC to obtain YYH-23 with a yield of 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] + Calculate for C36 H 36 FN2O 11 S,707.2069; observed 707.2059.

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

[0046] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-(4-phenyl)-1,3,4-oxadiazole, 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] + Calculate for C 36 H 37 N2O 11 ,673.2391; observed 673.2377.

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

[0049] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-oxadiazole, and YYH-2 was purified 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] + Calculate for C 36 H 36 FN2O 11 ,691.2297; observed 691.2291.

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

[0052] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole, 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] + Calculate for C 36 H 36 ClN2O 11 ,708.2080; observed708.2070.

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

[0055] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-(4-nitrophenyl)-1,3,4-oxadiazole, and YYH-5 was purified by high performance liquid chromatography as 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] + Calculate for C 36 H 36 N3O 13 ,718.2242; observed718.2235.

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

[0058] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-(3-nitrophenyl)-1,3,4-oxadiazole, and YYH-6 was purified 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] + Calculate for C 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(4-nitrophenyl)vinyl]-1,3,4-oxadiazole, and YYH-7 was purified 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(4-trifluoromethylphenyl)vinyl]-1,3,4-oxadiazole, and YYH-9 was purified 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(4-fluorophenyl)vinyl]-1,3,4-oxadiazole, and YYH-11 was purified 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(3-trifluoromethylphenyl)vinyl]-1,3,4-oxadiazole, and YYH-12 was purified 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(4-chlorophenyl)vinyl]-1,3,4-oxadiazole, and YYH-13 was purified 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(3-chlorophenyl)vinyl]-1,3,4-oxadiazole, and YYH-14 was purified 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(4-bromophenyl)vinyl]-1,3,4-oxadiazole, and YYH-15 was purified 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(4-methoxyphenyl)vinyl]-1,3,4-oxadiazole, and YYH-17 was purified 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(3-(dimethylamino)phenyl)vinyl]-1,3,4-oxadiazole, and YYH-19 was purified 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-[(1E)-2-(3-(dioxymethylene)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] + Calculate 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 step was replaced by 5-(3-{4-[bis(2-chloroethyl)amino]phenyl}propyl)-2-(chloromethyl)-1,3,4-oxadiazole, and YYH-22 was purified 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] + Calculate for C 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 step was replaced by 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-thiadiazole, and YYH-24 was purified 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-(4-bromophenyl)-1,3,4-thiadiazole, and YYH-25 was purified by high performance liquid chromatography as a yellow solid 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] + Calculate for C 36 H 36 BrN2O 10 S,767.1268; observed 767.1255.

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

[0115] According to the method in Example 2, 2-(chloromethyl)-5-(4-fluorophenyl)-1,3,4-thiadiazole in the above step was replaced by 2-(chloromethyl)-5-(4-trifluoromethylphenyl)-1,3,4-thiadiazole, and YYH-26 was obtained by high performance liquid chromatography purification 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 2-(chloromethyl)-5-(3-chlorophenyl)-1,3,4-thiadiazole, and YYH-28 was obtained by high performance liquid chromatography purification 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 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] + Calculate 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 step was replaced by 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] + Calculate for C 35 H 41 N2O 11 ,665.2705; observed 665.2681.

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

[0133] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-n-octyl-1,3,4-oxadiazole, and YYH-32 was obtained by HPLC purification 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] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-cyclopropyl-1,3,4-oxadiazole, and YYH-33 was obtained by HPLC purification as 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] + Calculate for C 33 H 37 N2O 11 ,637.2392; observed637.2368.

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

[0139] According to the method in Example 1, 2-(chloromethyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole in the above step was replaced by 2-(chloromethyl)-5-cyclohexyl-1,3,4-oxadiazole, and YYH-34 was obtained by HPLC purification 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] + Calculate for C 35 H 41 N2O 11 ,679.2861; observed.679.2838.

[0141] Example 35: Anti-liver cancer activity screening

[0142] In order to prove that the oxadiazole derivatives and thiadiazole derivatives YYH with icariin II as the parent core of the present invention have the potential to treat liver cancer, the compounds prepared above were screened for liver cancer pharmacodynamics. The specific screening method and results of anti-liver cancer compounds are as follows:

[0143] The MTT method was used for the initial screening of compounds, with icariin, ICA-II and sorafenib (SORA) as positive controls. Hepatocellular carcinoma cells or normal hepatocytes with good growth were selected, digested with trypsin, dispersed into single cell suspension with complete cell culture medium, and counted the cells. The cell concentration was adjusted to 5×10 4 / mL, gently blow the cells to make them evenly dispersed, evenly spread them in a 96-well plate (100μL / well), and culture them in a CO2 incubator overnight. The next day, the culture medium was discarded, and 0.78125, 1.5625, 3.125, 6.25, 12.5, and 25μM of the drugs to be tested were added, and a blank group was set to deduct the background effect, with 3 replicates in each group. After each group of samples was in the CO2 incubator for 72 hours, the 96-well plate was taken out and incubated in the incubator for 4 hours with the pre-prepared MTT solution, 20μL / well. Then the MTT solution was discarded, 150μL DMSO solution was added to each well to dissolve the sample, and the crystals were fully dissolved by shaking at room temperature for 5 minutes. The microplate reader was adjusted to OD 490nm to detect the absorbance value of each well, and the experiment was repeated 3 times. The cell growth inhibition rate was calculated according to the following formula: Inhibition rate (%) = (A control - A drug addition) / (A control - A blank) × 100%. Calculate the inhibition rate at different concentrations and draw a dose-effect curve. Calculate the concentration at which the growth inhibition rate is 50%, which is the IC 50 The results showed that most of the compounds had considerable activity in inhibiting the proliferation of liver cancer cells, as shown in Table 1.

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

[0145]

[0146]

[0147] In summary, by applying the above-mentioned pharmacological experiments to screen anti-tumor drugs for liver cancer cell proliferation, it was found that compounds such as YYH-7, 9, 11, 17, and 21 had much better inhibitory activity against liver cancer cells HepG2 than the positive control drugs (Acoradine, SORA, and ICA-II), and had value for further development. Therefore, the diazole heterocyclic derivatives of icariin II in the present invention had good anti-liver cancer cell proliferation activity and had the potential to be further developed into drugs for the treatment of liver cancer.

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

Claims

1. An icariin II derivative or a pharmaceutically acceptable salt thereof having anti-hepatocarcinoma activity, characterized in that: 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); the icariin II thiadiazole derivative has a chemical structure shown in formula (III) or formula (IV): R in the formula (I), (II), (III) or (IV) is selected from:

2. The icariin II derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The icariin II oxadiazole derivative or icariin II thiadiazole derivative is selected from the following compounds:

3. The method for preparing the icariin II derivative or a pharmaceutically acceptable salt thereof according to claim 1 or claim 2, characterized in that: The steps include: Using a1-a22 and a31-a34 as starting materials, step (i) is performed to obtain intermediates b1-b22 and b31-34, and then step (ii) is performed to obtain fragments c1-c22 and c31-34, and finally step (iii) is performed to replace icariin II to obtain compounds YYH1-22 and YYH31-34; Using a23-a30 as starting materials, step (i) is used to obtain intermediates b23-b30, and step (iv) is used to obtain intermediates c23-c30. Then, step (v) is used to obtain fragment d23-30. Finally, step (iii) is used to replace icariin II to obtain compound YYH23-30.

4. The preparation method according to claim 3, characterized in that: The preparation method comprises the following steps: (i) Synthesis of b1-b34: a1-a34 is dissolved in a solvent, 1.5-3 equivalents of HOBt and 1.5-3 equivalents of EDCI are added, stirred for 0.5-4 hours, 2-4 equivalents of 80% hydrazine hydrate are added dropwise, reacted at room temperature, the solvent is removed, extracted, washed, dried, filtered, and purified; (ii) Synthesis of c1-c22 and c31-34: dissolve b1-b22 and b31-34 in a solvent, add 1-3 equivalents of trimethoxyethyl chloride and 3-5 equivalents of acetic acid, react at 110-130°C, and post-treat in the same manner as step (i); (iii) Synthesis of YYH1-34: Dissolve icariin II, 1-2 equivalents of K2CO3, and 0.5-1 equivalents of KI in a solvent, stir at room temperature, add the corresponding fragment, react at 60°C, and post-treat in the same manner as step (i); (iv) Synthesis of d23-d30: b23-b30 was dissolved in a solvent, 1-2 equivalents of p-chloroacetic acid, 1.5-3 equivalents of HOBt and 1.5-3 equivalents of EDCI were added, and the reaction was carried out at room temperature. The post-treatment method was the same as step (i); (v) Synthesis of e23-e30: Dissolve d23-d30 in a solvent, add 2-3 equivalents of Lawesson's reagent, react at 110-130°C, and post-treat in the same manner as step (i).

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

6. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the icariin II derivative according to 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 injection dosage form.

8. The pharmaceutical composition according to claim 7, characterized in that: The oral dosage form is capsule, tablet, granule, oral solution, sustained-release preparation or controlled-release preparation, and the pharmaceutically acceptable carrier is selected from one or more of diluent, lubricant, binder, disintegrant, stabilizer or solvent.

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

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