Cordycepin derivatives with antitumor effects

Abstract

The present invention discloses a compound having an antitumor effect by derivatization of cordycepin, the structure of which is represented by formula I. The cordycepin derivative and its pharmaceutical composition according to the present invention have excellent antitumor growth effect. Compared with the parent drug, the cordycepin derivative has excellent affinity to cell membranes, so that the half-life during which the drug is metabolized in the body is longer, and the time during which the drug remains in the body is longer. Compared with other nucleoside antitumor drugs, the cordycepin derivative and its pharmaceutical composition according to the present invention have a wider range of tumor types and action, and have excellent inhibitory effect against gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, colorectal cancer, melanoma, ovarian cancer, etc., with reduced side effects and excellent therapeutic effect. JPEG2025511448000109.jpg42166

Description

[Technical field]

[0001] The present invention belongs to the field of biomedicine, specifically to a compound by derivatization of cordycepin and its preparation method, and its use in the preparation of products for preventing and treating related diseases caused by mutations of cell dysfunction. [Background technology]

[0002] With the accumulation of external environmental stimuli and cell damage, the cytocompatibility in the body changes. One is the loss of cytocompatibility, which eventually develops into aging, and the other is the abnormal enhancement of cytocompatibility, which transforms into cancer. Cancer is a common disease that threatens human life and health, and the incidence and mortality of cancer are showing an upward trend all over the world. At present, the treatment of malignant tumors mainly includes surgery, radiation therapy, and chemotherapy, and chemotherapy is mainly using synthetic drugs. The inhibitory effect of chemotherapy drugs on tumors is positive, and it is one of the effective and commonly used treatments for malignant tumors, but its toxic side effects are widespread and serious, and there is a problem of drug resistance. In addition, chemotherapy drugs have low selectivity for tumor cells and normal cells, and while they kill or inhibit tumor cells, they also damage the proliferation of normal cells, directly affect the functions of the heart, liver, kidneys, and nervous system, and cause certain toxicity to the human body. Therefore, it is very necessary to find low-toxicity and efficient antitumor drugs in tumor treatment.

[0003] Nucleosides, as one of the most important endogenous compounds in the human body, play an important role in the metabolic process of the body. Modification and derivatization of nucleoside compounds is one of the main methods for preparing antitumor drugs, and currently available nucleoside antitumor drugs include forodesine, fludarabine, cladribine, clofarabine, fludarabine phosphate, troxacitabine, etc. Cordycepin (3'-deoxyadenosine) is the main active ingredient of Cordyceps sinensis, which belongs to the nucleoside analogues and has excellent effects on cytocompatibility changes (anti-aging, anti-cancer), immune regulation, and inflammation elimination in the metabolic process of the body.

[0004] The mechanism of the anticancer effect of cordycepin is mainly to suppress the invasion and metastasis of tumor cells by inducing apoptosis, regulating the cell cycle, and interfering with the expression of matrix metalloproteinases (MMPs). Signal transduction pathways related to the induction of apoptosis in tumor cells include the NF-κB signal transduction pathway and the mitogen-activated protein kinase (MAPK) signal transduction pathway. Regulation of the cell cycle is mainly due to cordycepin shortening the G1 phase of the cell cycle in cancer cells, prolonging the G2 and M phases, causing the cell cycle to remain in the G2 / M phase, and inhibiting cell proliferation. The process of interfering with the expression of matrix metalloproteinases mainly inhibits the NF-κB signal transduction pathway, and ultimately inhibits the expression of MMP-9. However, nucleoside analogues have poor lipid solubility, are difficult to absorb, are easily metabolically inactivated by deaminases, have short half-lives, weak targeting properties, and are prone to drug resistance in some tumor cells and viruses, all of which greatly reduce the effectiveness of the use of nucleoside drugs. Summary of the Invention [Problem to be solved by the invention]

[0005] Objective of the present invention: In view of the shortage of conventional nucleoside drugs, the technical problem to be solved by the present invention is to provide a series of nucleoside analogues (cordycepin derivatives) that have been chemically modified and have cordycepin as the base nucleus, while retaining the natural pharmacological activity of cordycepin.

[0006] A further technical problem to be solved by the present invention is to provide a composition containing the above-mentioned nucleoside analogue.

[0007] Furthermore, the technical problem to be solved by the present invention is to provide the use of said nucleoside analogues and compositions thereof in products for preventing and treating diseases associated with mutations in cellular dysfunction in mammals or humans.

[0008] Finally, the technical problem to be solved by the present invention is to provide a method for preparing the above-mentioned nucleoside analogues. [Means for solving the problem]

[0009] In order to solve the above first technical problem, the present invention discloses a cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka]

[0010] In the formula, R1 is selected from hydrogen, a phosphoric acid group, a substituted phosphoric acid ester group, a phosphonic acid group, a substituted phosphonic acid ester group, an alkyl alcohol group, an amino acid alkyl ester group, an amino acid alkyl alcohol ester group, an alkyl acid alkyl ester group, or a cycloalkyl polyol group, and the substitution is substituted with one or more functional groups selected from an alkoxy group, an alkoxy group substituted with a halogen, an aryloxy group, an amide amino acid group, an alkyl ester group, and an alkyl acid methyl ester oxy group; R2 is selected from hydrogen or an azide group; R3 is selected from hydrogen, fluorine, chlorine, or an azide group; R4 is a hydroxyl group, a cyano group, a β-amide-γ-cyclohexane group, a ... R5 is selected from hydrogen, a bromovinyl group, a mercapto group, a methyl group, fluorine, or chlorine; R6 is selected from an amino group or a substituted carboxamide group, the substitution being with one or more functional groups selected from an alkyl group, an aryl group, a cycloalkyl group, a furyl group, and a pyridyl group; R7 is selected from hydrogen or an isopropylamino group; and R1, R2, R3, R5, and R7 are selected from hydrogen, R4 is selected from a hydroxyl group, and R6 is selected from an amino group, and is not present.

[0011] In some embodiments, R1 is hydrogen, R1 1-R1 7 a phosphate group represented by the formula: diethyl phosphate group, di-n-propyl phosphate, diisopropyl phosphate, diisobutyl phosphate, di-n-butyl phosphate, or diisobutyryloxymethoxy phosphate; 8 -R1 11 phosphonic acid group represented by the formula: ethyl trifluorophosphonate, diisobutyryloxymethoxy phosphonate, or pivaloyloxymethoxy phosphonate, or R 12 -R1 13 a cyclophosphonotrifluoroglycol ester group represented by the formula: 14 -R1 16 (R)-phosphoryl isoleucine methyl ester phenyl ester, (S)-phosphoryl isoleucine methyl ester phenyl ester, or phosphoalanine isopropyl ester phenyl ester represented by the formula: 17 -R1 22 Hydroxyethyl, 1,3-dihydroxy-2-propyl, alanine ethyl ester, isoleucine ethyl ester, isoleucine-1,3-propanediol ester, or glycine ethyl ester, represented by the formula: 23 -R1 25 In some embodiments, R1 is selected from dimethyldicarbonate-propylene glycol, 2-isopropoxyhexahydrocyclopentane[d][1,3,2]dioxolin-7-ol, or 1-hydroxymethyl-2,3-dihydroxy-4-cyclopentyl, represented by the formula: 1 , R1 5 , R1 8 , R1 10 , R1 15 , R1 16 , R1 17 , R1 21 The structure is selected from any one of the structures represented by: [ka]

[0012] In some embodiments, R4 is 1-R4 7 In some embodiments, R4 is selected from a hydroxyl group, a cyano group, a β-amido-γ-cyclosulfonyloxy, a 2-amino-propionyloxy, a 2-amino-3-methyl-butyryloxy, a phosphoryloxyalanine methyl ester phenyl ester, or a phosphonamide isoleucine methyl ester phenyl ester represented by the formula: 3 , R4 5 , or R4 6 The structure is selected from any one of the structures represented by: [ka]

[0013] In some embodiments, R5 is selected from hydrogen, a bromovinyl group, a mercapto group, fluorine, or chlorine.

[0014] In some embodiments, R6 is an amino group, or R6 1 -R6 16 In some embodiments, R6 is selected from an acetamido group, a butylamido group, an octylamido group, a dodecylamido group, an octadecylamido group, an isopropionamido group, an isobutylamido group, a pivalamido group, a 2-ethyl-n-butylamido group, a 3,3-dimethyl-butylamido group, a cyclohexylcarboxamido group, a cyclopentylcarboxamido group, a benzamido group, a furancarboxamido group, a pyridinecarboxamido group, or a hexadecylamido group, 5 , R6 12 , R6 13 , or R6 15 The structure is selected from any one of the structures represented by: [ka]

[0015] In some embodiments, the cordycepin derivative is represented by formula I, or a pharma- ceutically acceptable salt thereof, wherein R1 is R1 16wherein R2 is selected from hydrogen, R3 is selected from hydrogen, R4 is selected from a hydroxyl group, R5 is selected from hydrogen, fluorine or chlorine, R6 is selected from a pyridinecarboxamide or an amino group, and R7 is selected from hydrogen. In some embodiments, the cordycepin derivative represented by formula I is selected from compound 24 below, compound 7 below, or compound 20 below.

[0016] In some embodiments, the cordycepin derivative represented by formula I is selected from any one of compounds 1 to 40. [ka] [ka]

[0017] In order to solve the second technical problem, the present invention discloses a pharmaceutical composition comprising at least one of the cordycepin derivatives, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof, and at least one immune checkpoint inhibitor.

[0018] In some embodiments, the immune checkpoint inhibitor is selected from PD-1 and / or CTLA4 monoclonal antibodies.

[0019] In some embodiments, the mass ratio of the cordycepin derivative, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof to the immune checkpoint inhibitor is 1:0.2-10, in some embodiments, 1:0.2-8, in some embodiments, 1:0.2-5, in some embodiments, 1:0.2-3, in some embodiments, 1:0.2-2, in some embodiments, 1:0.5-1.5, in some embodiments, 1:0.8-1.2, and in some embodiments, 1:1.

[0020] In some embodiments, the dosage form of the pharmaceutical composition is selected from a tablet, a pellet, a capsule, a pill, a syrup, a disintegrant, an injection, a sustained release formulation, or a kit.

[0021] In order to solve the third technical problem, the present invention discloses the use of the above cordycepin derivative, or its pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof, or the above pharmaceutical composition, in the preparation of a product for preventing and treating associated diseases caused by mutations in cell dysfunction in mammals or humans.

[0022] In some embodiments, the disease associated with mutations in the cellular dysfunction is a tumor, and in some embodiments, the tumor includes, but is not limited to, gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, colorectal cancer, esophageal cancer, prostate cancer, melanoma, glioma, and ovarian cancer, and in some embodiments, the tumor is any one of gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, colorectal cancer, melanoma, and ovarian cancer.

[0023] In some embodiments, the product includes, but is not limited to, a pharmaceutical product.

[0024] In order to solve the above-mentioned fourth technical problem, the present invention discloses a preparation method for the above-mentioned cordycepin derivatives, and the preparation method is a synthetic means corresponding to a modification method. In consideration of the fact that other active groups in cordycepin or modified cordycepin molecules are involved in the reaction during the reaction, the active groups should be properly protected, the protection method of the present invention is the protection and deprotection of hydroxyl groups and amino groups, and the protection and deprotection means are all conventional means in the art, and some other reaction systems need to be protected by adding protective gas, which are all conventional means in the experimental process in the art.

[0025] Specifically, the following are included:

[0026] A. Qualify R1 In formula I, R1 is selected from a phosphate group, a substituted phosphate ester group, a phosphonic acid group, a substituted phosphonic acid ester group, an alkyl alcohol group, an amino acid alkyl ester group, an amino acid alkyl alcohol ester group, an alkyl acid alkyl ester group, or a cycloalkyl polyol group, and the substitution is substituted with one or more functional groups selected from an alkoxy group, an alkoxy group substituted with a halogen, an aryloxy group, an amide amino acid group, an alkyl ester group, and an alkyl acid methyl ester oxy group. In this case, the cordycepin derivative represented by formula I is prepared by chemically reacting compound IR-1 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I. [ka]

[0027] In formula IR-1, R2 to R7 are the same as R2 to R7 in formula I, or each is independently selected from a protecting group.

[0028] When R1 in formula I is selected from a phosphate group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-1 with a modifying agent, phosphorus oxychloride, in trimethyl phosphite and / or triethyl phosphite. In some embodiments, the ratio of the amounts of compound IR-1, the modifying agent and the organic solvent used is 1 mmol: 3-6 mmol: 15-30 mL, in some embodiments, the reaction temperature is -10-5°C, and in some embodiments, the reaction time is 0.5-2 hours.

[0029] When R1 in formula I is selected from a substituted phosphate ester group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-1 with nitrophenyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide and / or tetrahydrofuran using tert-butylmagnesium chloride as a catalyst, the substitution being substituted with one or more functional groups selected from an alkoxy group, an alkoxy group substituted with a halogen, an aryloxy group, an amide amino acid group, an alkyl ester group, and an alkyl acid methyloxy group; in some embodiments, the ratio of the amounts of compound IR-1, the modifying agent, the catalyst, and the organic solvent used is 1 mmol: 1-3 mmol: 1-2 mmol: 9-15 mL; in some embodiments, the reaction temperature is 20-40° C.; and in some embodiments, the reaction time is 2-5 hours.

[0030] In some embodiments, when R1 in formula I is selected from a phosphonic acid group or a substituted phosphonic acid ester group, the method for preparing the cordycepin derivative represented by formula I is to react compound IR-1 with p-toluenesulfonyloxymethyl phosphate substituted or unsubstituted with a modifying agent in anhydrous N,N-dimethylformamide using NaH as a catalyst, and the substitution is an alkoxy group, an alkoxy group substituted with a halogen, an aryloxy group, an amide amino acid group, an alkyl ester group, an alkyl acid methyl group, an alkyl ester ... In some embodiments, the ratio of the amounts of compound IR-1, modifying agent, catalyst, and organic solvent used is 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:100-200 mL, and in some embodiments, 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:150 mL, in some embodiments, the reaction temperature is -20 to 0°C, and in some embodiments, the reaction time is 0.5 to 6 hours.

[0031] When R1 in formula I is selected from an alkyl alcohol group, an amino acid alkyl ester group, an amino acid alkyl alcohol ester group, an alkyl acid alkyl ester group, or a cycloalkyl polyol group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I includes reacting compound IR-1 with a modifying agent which is an alkyl alcohol, an amino acid alkyl ester, an amino acid alkyl alcohol ester, an alkyl acid alkyl ester, or a cycloalkyl polyol correspondingly substituted with a halogen in methyl ethyl ketone using potassium carbonate as a catalyst, in some embodiments, the halogen is bromine or chlorine, in some embodiments, the ratio of the amounts of compound IR-1, the modifying agent, the catalyst, and the organic solvent used is 1 mmol:0.5-1.5 mmol:1-3 mmol:5-8 mL, and in some embodiments, 1 mmol:1 mmol:1-3 mmol:5-8 mL, in some embodiments, the reaction temperature is 40-100° C., and in some embodiments, the reaction time is 8-20 hours.

[0032] B. Qualify R2 In the case where R2 in formula I is selected from an azide group, the method for preparing the cordycepin derivative represented by formula I includes carrying out a cyclization reaction of compound IR-2 as a raw material in an organic solvent to obtain intermediate IR-2a, 5-(6-amino-9H-purin-9-yl)-1,4-dioxapyrrole[2.4]heptan-6-ol or a derivative thereof, and carrying out a ring-opening reaction of the obtained intermediate IR-2a in an organic solvent to obtain the cordycepin derivative represented by formula I. [ka]

[0033] In formula IR-2, R1, R3 to R7 are the same as R1, R3 to R7 in formula I, or each is independently selected from a protecting group.

[0034] In some embodiments, the method for preparing the intermediate IR-2a comprises subjecting compound IR-2 to a cyclization reaction in dichloromethane under the catalysis of diphosphorus pentoxide and metachloroperbenzoic acid; in some embodiments, the ratio of the amounts of compound IR-2, diphosphorus pentoxide, metachloroperbenzoic acid, and dichloromethane used is 1 mmol: 1.2-2 mmol: 2-3 mmol: 10-20 mL; in some embodiments, the cyclization reaction temperature is 20°C to 60°C; and in some embodiments, the cyclization reaction time is 3 hours to 10 hours.

[0035] In some embodiments, the method for preparing the cordycepin derivative represented by formula I includes subjecting the obtained intermediate IR-2a to a ring-opening reaction with sodium azide in anhydrous dimethylformamide; in some embodiments, the ratio of the amounts of the compound IR-2a, sodium azide, and dimethylformamide used is 1 mmol: 4-5 mmol: 2-5 mL; in some embodiments, the ring-opening reaction temperature is 100-120°C; and in some embodiments, the ring-opening reaction time is 12-16 hours.

[0036] C. Qualify R3 In the case where R3 in formula I is selected from fluorine, chlorine, or azide group, the method for preparing the cordycepin derivative represented by formula I is to chemically react compound IR-3 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I. [ka]

[0037] In formula IR-3, R1, R2, R5 to R7 are the same as R1, R2, R5 to R7 in formula I, or each is independently selected from protecting groups.

[0038] In the case where R3 in formula I is selected from fluorine or chlorine, in some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-3 with trifluoromethanesulfonic anhydride in pyridine and dichloromethane to obtain intermediate IR-3a, 5-(6-amino-9H-purin-9-yl)-4-hydroxy-2-(hydroxymethyl)tetrahydrofuran-3-yl trifluoromethanesulfonate or its derivative, and then in ethyl acetate, displacing intermediate IR-3a with hydrofluoric acid, sulfur trifluoride or hydrochloric acid to obtain the cordycepin derivative represented by formula I. In some embodiments, the method for preparing the intermediate IR-3a comprises using the compound IR-3, trifluoromethanesulfonic anhydride, pyridine and dichloromethane in a ratio of 1 mmol:1 to 1.5 mmol:0. In some embodiments, the reaction temperature is -5 to 5°C, and in some embodiments, the reaction time is 1 to 3 hours. In some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises subjecting intermediate IR-3a to a substitution reaction with a triethylamine solution of 37% hydrofluoric acid, diethylaminosulfur trifluoride, or hydrochloric acid, and in some embodiments, the ratio of the amounts of intermediate IR-3a, 37% hydrofluoric acid, diethylaminosulfur trifluoride, or hydrochloric acid in the triethylamine solution, and ethyl acetate used is 1 mmol:2-3 mmol:4-10 mL. In some embodiments, the reaction temperature is 60 to 80°C, and in some embodiments, the reaction time is 8 to 10 hours.

[0039] In the case where R3 in formula I is selected from an azide group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I includes: in N,N-dimethylformamide, compound IR-3 is cyclized under the catalysis of triphenylphosphine and diisopropyl azodicarboxylate to obtain intermediate IR-3b, 4-(6-amino-9H-purin-9-yl)-3,6-dioxane[3.1.0]hexan-2-yl)methanol or a derivative thereof; and in dimethylformamide, the obtained intermediate IR-3b is ring-opened with sodium azide to obtain the cordycepin derivative represented by formula I. In some embodiments, the method for preparing the intermediate IR-3b includes: in N,N-dimethylformamide, compound IR-3, triphenylphosphine, diisopropyl azodicarboxylate, and N, The ratio of the amounts of N-dimethylformamide used is 10.0 mmol: 16-28 mmol: 1.6-2.8 mmol: 15-50 mL, and in some embodiments, 10.0 mmol: 22 mmol: 2.2 mmol: 15-50 mL. In some embodiments, the cyclization reaction temperature is 10-60°C. In some embodiments, the cyclization reaction time is 1-5 hours. In some embodiments, in the method for preparing the cordycepin derivative represented by formula I, the ratio of the amounts of intermediate IR-3b, sodium azide, and dimethylformamide used is 1 mmol: 4-5 mmol: 2-5 mL. In some embodiments, the ring-opening reaction temperature is 100-120°C. In some embodiments, the ring-opening reaction time is 12-16 hours.

[0040] D. Qualify R4 In the case where R4 in formula I is selected from the group consisting of a cyano group, a β-amide-γ-cyclosulfonyloxy group, an amino acid carboxylate group, an amino acid alkyl ester phosphonic acid phenyl ester group, and an amino acid alkyl ester phosphate phenyl ester group, the method for preparing the cordycepin derivative represented by formula I comprises chemically reacting compound IR-4 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I. [ka]

[0041] In formula IR-4, R1 to R3 and R5 to R7 are the same as R1 to R3 and R5 to R7 in formula I, or each is independently selected from protecting groups.

[0042] In some embodiments, when R4 in formula I is selected from a cyano group, the method for preparing the cordycepin derivative represented by formula I includes stirring compound IR-4, trifluoromethanesulfonic acid, and trimethylsilyl triflate in dichloromethane at −50 to −30° C., and then reacting with trimethylsilyl cyanide and triethylamine. In some embodiments, the ratio of the amounts of the compound IR-4, trifluoromethanesulfonic acid, trimethylsilyl triflate, and dichloromethane used is 10 mmol: 0.8-1.8 mL: 2.2-3.2 mL: 90-110 mL, and in some embodiments, 10 mmol: 1.3 mL: 2.7 mL: 100 mL. In some embodiments, the stirring time is 20-40 minutes, and in some embodiments, 30 minutes. In some embodiments, the ratio of the amounts of the compound IR-4, trimethylsilyl cyanide, and triethylamine used is 10 mmol: 3.4-4.3 g: 3-4 mL. In some embodiments, the reaction temperature is 20-30° C., and in some embodiments, room temperature. In some embodiments, the reaction time is 2-4 hours.

[0043] In the case where R4 in formula I is selected from β-amido-γ-cyclosulfonyloxy groups, in some embodiments, the method for preparing the cordycepin derivative represented by formula I includes oxidizing compound IR-4 to obtain intermediate IR-4a, 2-(6-amino-1,6-dihydro-9H-purin-9-yl)-5-(hydroxymethyl)dihydrofuran-3(2H)-one or its derivative, and then cyanating the intermediate IR-4b, 2-(6-amino-1,6-dihydro 1,6-dihydro-9H-purin-9-yl)-5-(hydroxymethyl)-3-isocyanatotetrahydrofuran-3-ol or a derivative thereof is obtained, which is then methylsulfonated to obtain intermediate IR-4c, 2-(6-amino-1,6-dihydro-9H-purin-9-yl)-5-(hydroxymethyl)-3-isocyanatotetrahydrofuran-3-yl methanesulfonate or a derivative thereof, which is then ring-closed to obtain the cordycepin derivative represented by formula I. In some embodiments, the preparation method of intermediate IR-4a comprises reacting compound IR-4 with dichromium trioxide of Jones reagent (2.2M) in acetone, in some embodiments, the ratio of the amounts of compound IR-4, Jones reagent and acetone used is 20mmol:5-8mL:50-100mL, in some embodiments, the reaction temperature is 0-40°C, and in some embodiments, the reaction time is 1-4 hours. In some embodiments, the method for preparing the intermediate IR-4b comprises reacting the intermediate IR-4a with trimethylsilyl cyanide and boron trifluoride diethyl etherate in dichloromethane; in some embodiments, the ratio of the amounts of the intermediate IR-4a, trimethylsilyl cyanide, boron trifluoride diethyl etherate and dichloromethane used is 20 mmol: 20-40 mmol: 10-30 mmol: 50-100 mL, or in some embodiments, 20 mmol: 20-40 mmol: 20 mmol: 50-100 mL; in some embodiments, the reaction temperature is 0-40°C; and in some embodiments, the reaction time is 1-4 hours.In some embodiments, the method for preparing the intermediate IR-4c comprises reacting the intermediate IR-4b with triethylamine and methanesulfonyl chloride in anhydrous dichloromethane; in some embodiments, the ratio of the amounts of the intermediate IR-4b, triethylamine, methanesulfonyl chloride, and anhydrous dichloromethane used is 2.2 mmol: 10-15 mmol: 4-8 mmol: 5-10 mL; in some embodiments, the reaction temperature is −30 to 0° C.; and in some embodiments, the reaction time is 2 to 4 hours. In some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting intermediate IR-4c with cesium carbonate in anhydrous acetonitrile, in which the ratio of the amounts of intermediate IR-4c, cesium carbonate and anhydrous acetonitrile used is 1 mmol:0.5-2.5 mmol:3-10 mL, or in some embodiments, 1 mmol:1.5 mmol:3-10 mL, in which the reaction temperature is 0-40° C., and in which the reaction time is 2-4 hours.

[0044] In the case where R4 in formula I is selected from an amino acid carboxylate group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-4 with amino acid chloride in anhydrous pyridine to obtain the cordycepin derivative represented by formula I. In some embodiments, the ratio of the amounts of compound IR-4, amino acid chloride, and pyridine used is 10 mmol: 5-15 mmol: 50-100 mL, and in some embodiments, 10 mmol: 10 mmol: 50-100 mL; in some embodiments, the reaction temperature is 20-60°C, and in some embodiments, 40°C; and in some embodiments, the reaction time is 6-20 hours.

[0045] In the case where R4 in formula I is selected from an amino acid alkyl ester phosphonic acid phenyl ester group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-4 with p-toluenesulfonyloxymethylphenyl phosphate, which is a modifying agent, substituted with an amino acid alkyl ester, in anhydrous N,N-dimethylformamide using NaH as a catalyst. In some embodiments, the ratio of the amounts of compound IR-4, the modifying agent, NaH, and anhydrous N,N-dimethylformamide used is 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:100-200 mL, or 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:150 mL in some embodiments, the reaction temperature is -20-0°C, and in some embodiments, the reaction time is 0.5-6 hours.

[0046] In the case where R4 in formula I is selected from an amino acid alkyl ester phosphate phenyl ester group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-4 with nitrophenyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide and / or tetrahydrofuran using tert-butylmagnesium chloride as a catalyst, the substitution being substituted with one or more functional groups of an aryloxy group or an alkyl acid methyloxy group. In some embodiments, the ratio of the amounts of compound IR-4, modifying agent, catalyst, and organic solvent used is 1 mmol: 1-3 mmol: 1-2 mmol: 9-15 mL, in some embodiments, the reaction temperature is 20-40° C., and in some embodiments, the reaction time is 2-5 hours.

[0047] Qualify E.R5 In the case where R5 in formula I is selected from a bromovinyl group, a mercapto group, a methyl group, fluorine, or chlorine, the method for preparing the cordycepin derivative represented by formula I is to obtain the cordycepin derivative represented by formula I by chemically reacting compound IR-5 as a raw material in an organic solvent. [ka]

[0048] In formula IR-5, R1 to R4 and R6 to R7 are the same as R1 to R4 and R6 to R7 in formula I, or each is independently selected from protecting groups.

[0049] In the case where R5 in formula I is selected from a bromovinyl group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I includes iodinating compound IR-5 to obtain intermediate IR-5a, 2-(6-amino-2-iodo-1,6-dihydro-9H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3-ol or its derivative, and then substituting intermediate IR-5a with methyl acrylate to obtain intermediate IR-5b, methyl(E)-3-(6-amino-3-hydroxy-5H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3-ol or its derivative. 3-(6-amino-9-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-6,9-dihydro-1H-purin-2-yl)acrylate or its derivative, intermediate IR-5b is hydrolyzed to obtain intermediate IR-5c, (E)-3-(6-amino-9-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-6,9-dihydro-1H-purin-2-yl)acrylic acid or its derivative, intermediate IR-5c is brominated with N-bromosuccinimide to obtain cordycepin derivative represented by formula I. In some embodiments, the method for preparing intermediate IR-5a includes iodination of compound IR-5 with elemental iodine in dilute nitric acid to obtain intermediate IR-5a. In some embodiments, the ratio of the amount of the compound IR-5 to the amount of elemental iodine used is 1 mmol:0.5-0.8 mmol, in some embodiments, the reaction temperature is 100-120° C., in some embodiments, 110° C., and in some embodiments, the reaction time is 4-6 hours. In some embodiments, the method for preparing the intermediate IR-5b comprises reacting the intermediate IR-5a with methyl acrylate and triethylamine in 1,4-dioxane under the catalysis of palladium acetate and triphenylphosphine.In some embodiments, the ratio of the amounts of the intermediate IR-5a, methyl acrylate, triethylamine, palladium acetate, triphenylphosphine, and 1,4-dioxane used is 1 mmol: 3-4 mmol: 0.1-0.5 mL: 0.01-0.09 mmol: 0.05-0.15 mmol: 10-20 mL, and in some embodiments, 1 mmol: 3-4 mmol: 0.1-0.5 mL: 0.05 mmol: 0.01 mmol: 10-20 mL, in some embodiments, the reaction temperature is 50-90° C., and in some embodiments, the reaction time is 0.5-2 hours. In some embodiments, the method for preparing the intermediate IR-5c comprises hydrolyzing the intermediate IR-5b with a sodium hydroxide solution. In some embodiments, the concentration of the sodium hydroxide solution is 0.5-3.5 mol / L, and in some embodiments, 2 mol / L. In some embodiments, the ratio of the amount of the intermediate IR-5b to the amount of the sodium hydroxide solution is 1 g:10-14 mL, and in some embodiments, 1 g:12 mL. In some embodiments, the reaction temperature is 20-30° C., and in some embodiments, room temperature. In some embodiments, the reaction time is 3-5 hours. In some embodiments, the method for producing the cordycepin derivative represented by formula I comprises reacting the intermediate IR-5c with N-bromosuccinimide under potassium carbonate catalyst in a mixed solvent of water and acetone. In some embodiments, the ratio of the amount of the intermediate IR-5c, N-bromosuccinimide, potassium carbonate, and mixed solvent is 1 mmol:1-3 mmol:1-3 mmol:15-30 mL. In some embodiments, the volume ratio of water to acetone is 1:4-8, and in some embodiments, 1:6. [ka]

[0050] In some examples, unmodified cordycepin was directly modified with bromovinyl, the reaction pathway being: [ka] It is.

[0051] In the case where R5 in formula I is selected from mercapto group, in some embodiments, the method for preparing the cordycepin derivative represented by formula I includes reacting compound IR-5 with hydrogen peroxide in acetic acid to obtain intermediate IR-5d, 1N-oxide-3'-deoxyadenosine or its derivative, and heating intermediate IR-5d in hydrochloric acid aqueous solution to reflux to obtain intermediate IR-5e, 5-amino-N'-hydroxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2- IR-5e is dissolved in water and reacted with Raney nickel catalyst under hydrogen atmosphere to obtain intermediate IR-5f, 5-amino-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1H-imidazole-4-carboxamide or a derivative thereof, and intermediate IR-5f is reacted with methanol, pyridine and carbon disulfide to obtain cordycepin derivative represented by formula I. In some embodiments, the method for preparing intermediate IR-5d comprises reacting compound IR-5 with acetic acid in hydrogen peroxide solution. In some embodiments, the concentration of the hydrogen peroxide solution is 20% to 40%, and in some embodiments, the concentration of the hydrogen peroxide solution is 30%, and in some embodiments, the ratio of the amounts of the compound IR-5, the hydrogen peroxide solution, and the acetic acid used is 1 mmol: 2 to 3 mol: 1 to 6 mL, and in some embodiments, the reaction temperature is 30 to 50°C, and in some embodiments, the reaction time is 2 to 4 days. In some embodiments, the method for preparing the intermediate IR-5e comprises heating the intermediate IR-5d in an aqueous hydrochloric acid solution to reflux. In some embodiments, the concentration of the aqueous hydrochloric acid solution is 1 to 5 mol / L, and in some embodiments, 3 mol / L, and in some embodiments, the ratio of the amounts of the intermediate IR-5d and the aqueous hydrochloric acid used is 1 mmol: 3 to 5 mL, and in some embodiments, the heating to reflux time is 10 to 30 minutes.In some embodiments, the ratio of the amounts of the intermediate IR-5e, Raney nickel, and water used is 1 mmol: 0.08-0.2 g: 10-20 mL, the reaction temperature is 50-70° C., and the reaction time is 2-5 days. In some embodiments, the ratio of the amounts of the intermediate IR-5f, methanol, pyridine, and carbon disulfide used is 1 mmol: 5-10 mL, the ratio of the amounts of the methanol, pyridine, and carbon disulfide used is 4: 3-7: 0.5-3.5, and in some embodiments, 4: 5: 2, the reaction temperature is 30-50° C., and in some embodiments, 40° C., and the reaction time is 3-5 days. [ka]

[0052] In some examples, unmodified cordycepin is directly modified with a sulfhydryl group, and the specific reaction method is as follows: [ka] It is.

[0053] In some embodiments, when R5 in formula I is selected from a methyl group, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-5, iodomethane, and potassium carbonate in dichloromethane solvent. In some embodiments, the ratio of the amounts of compound IR-5, iodomethane, potassium carbonate, and dichloromethane used is 1 mmol: 1-2 mmol: 1.5-3 mmol: 5-10 mL, in some embodiments, the reaction temperature is 20-50° C., and in some embodiments, the reaction time is 3-10 hours.

[0054] In the case where R5 in formula I is selected from fluorine or chlorine, in some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises nitro-derivating compound IR-5 to obtain nitrated intermediate IR-5g, 2-(6-amino-2-nitro-1,6-dihydro-9H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3-ol, which is then subjected to a substitution reaction to obtain the cordycepin derivative represented by formula I. In some embodiments, the method for preparing the intermediate IR-5g comprises reacting compound IR-5 with tetrabutylammonium nitrate in dichloromethane using trifluoroacetic anhydride as a catalyst. In some embodiments, the ratio of the amounts of compound IR-5, tetrabutylammonium nitrate, trifluoroacetic acid, and dichloromethane used is 1 mmol: 1.4-2 mmol: 1-2 mmol: 15-35 mL, in some embodiments, the reaction temperature is -10-10°C, and in some embodiments, the reaction time is 0.5-20 hours. In some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting 5 g of the nitrated intermediate IR-1 with tetrabutylammonium fluoride or tetrabutylammonium chloride in acetonitrile. In some embodiments, the ratio of the amounts of the nitrated intermediate IR-1, tetrabutylammonium fluoride or tetrabutylammonium chloride, and acetonitrile used is 1 mmol: 1.3-1.5 mmol: 30-50 mL, the reaction temperature is -5-5°C, and in some embodiments, 0°C, and in some embodiments, the reaction time is 20-30 minutes. [ka]

[0055] Qualify F.R6 In the formula I, when R6 is selected from a substituted carboxamide group, and the substitution is substituted with one or more functional groups including an alkyl group, an aryl group, a cycloalkyl group, a furyl group, and a pyridyl group, the cordycepin derivative represented by the formula I is prepared by chemically reacting compound IR-6 as a raw material in an organic solvent to obtain the cordycepin derivative represented by the formula I. [ka]

[0056] In formula IR-6, R1 to R5 and R7 are the same as R1 to R5 and R7 in formula I, or each is independently selected from a protecting group.

[0057] In some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-6 with a substituted acid chloride in anhydrous pyridine, the substitution being with one or more functional groups selected from the group consisting of an alkyl group, an aryl group, a cycloalkyl group, a furyl group, and a pyridyl group. In some embodiments, the ratio of the amounts of compound IR-6, the substituted acid chloride, and anhydrous pyridine used is 1 mmol: 1-2 mmol: 5-10 mL, in some embodiments, the reaction temperature is 0-60° C., and in some embodiments, the reaction time is 2-20 hours.

[0058] Qualifying G.R7 In the case where R7 in formula I is selected from isopropylamino group, the method for preparing the cordycepin derivative represented by formula I is to chemically react compound IR-7 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I. [ka]

[0059] In formula IR-7, R1 to R6 are the same as R1 to R6 in formula I, or each is independently selected from a protecting group.

[0060] In some embodiments, the method for preparing the cordycepin derivative represented by formula I comprises reacting compound IR-7 with 2-propylamine in dioxane. In some embodiments, the ratio of the amounts of compound IR-7, 2-propylamine and dioxane used is 10 mmol: 10-30 mmol: 50-100 mL, in some embodiments, the reaction temperature is 60-120° C., and in some embodiments, the reaction time is 10-30 hours.

[0061] In the present invention, protecting groups include, but are not limited to, -OTBS, -OAc, -NHCbz, -OTBPS, and -OTBDPS. In some embodiments, R4 is the same as R4 in formula I or is selected from -OTBS, -OAc, and -OTBDPS. In some embodiments, R6 is the same as R6 in formula I or is selected from -NHCbz.

[0062] In the present invention, the substituted nitrophenyl phosphate (substituted with one or more functional groups selected from the group consisting of alkoxy, halogen-substituted alkoxy, aryloxy, amide amino acid, alkyl ester, and alkyl acid methyloxy) can be obtained by the following method or other methods in the prior art.

[0063] In anhydrous dichloromethane, phenyl chlorophosphate, p-nitrophenol, and the corresponding substituted alcohol or amine are prepared with triethylamine as a catalyst under an environment of 0-25° C., and the substitution is substituted with one or more functional groups selected from alkoxy groups, halogen-substituted alkoxy groups, aryloxy groups, amide amino acid groups, alkyl ester groups, and alkyl acid methyloxy groups. In some embodiments, the molar volume ratio of the phenyl chlorophosphate, p-nitrophenol, the corresponding substituted product, triethylamine, and anhydrous dichloromethane is 1 mmol:1 mmol:1-2 mmol:2-5 mmol:5-10 mL.

[0064] In the present invention, the substituted p-toluenesulfonyloxymethyl phosphate (substituted with one or more functional groups selected from the group consisting of an alkoxy group, an alkoxy group substituted with a halogen, an aryloxy group, an amide amino acid group, an alkyl ester group, and an alkyl acid methyloxy group) can be obtained by the following method or other methods in the prior art.

[0065] The corresponding substituted chlorophosphate, p-toluenesulfonyl chloride, and formaldehyde are prepared in toluene under an environment of 0 to 105° C. with triethylamine as a catalyst. The substituted chlorophosphate is a chlorophosphate substituted with one or more functional groups selected from an alkoxy group, an alkoxy group substituted with a halogen, an aryloxy group, an amide amino acid group, an alkyl ester group, and an alkyl acid methyloxy group. In some embodiments, the molar volume ratio of the corresponding substituted chlorophosphate, p-toluenesulfonyl chloride, formaldehyde, triethylamine, and toluene is 1 mol:1 mol:0.8-1.2 mol:180-210 mL:500-800 mL.

[0066] As can be easily understood by those skilled in the art, the present invention is to modify the groups R1 to R7 based on cordycepin, but when R1 is hydrogen, R2 is hydrogen, R3 is hydrogen, R5 is hydrogen, R7 is hydrogen, R4 is a hydroxyl group, or R6 is an amino group, there is no need to modify them. In the preparation method of the present invention, each of the above-mentioned substituents can be modified according to the priority order of the stability of the intermediate prepared without going against the common knowledge in the art, thereby obtaining each embodiment of the present invention. For example, modifications not including ester groups, phosphate groups, and phosphonic acid groups (e.g., modifications with -CN, -N3, -F, -SH, etc.) can be performed first, followed by modification of ester groups, and finally modification of phosphate groups or phosphonic acid groups.

[0067] The term "prevention" in the present invention is meant to include administering a compound or formulation as described herein to prevent a disease or one or more symptoms associated with said disease, including preventing the occurrence of a disease or disease state in mammals, particularly cancer symptoms to which these mammals are susceptible.

[0068] The term "pharmacologically acceptable" in this invention means those compounds, materials, compositions and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals, within the scope of sound medical judgment, without undue toxicity, irritation, anaphylaxis or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0069] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic and other mixtures, such as mixtures enriched in enantiomers or diastereomers, all of which are within the scope of the present invention. Substituents such as alkyl groups may have additional asymmetric carbon atoms. All of these isomers and mixtures thereof are included within the scope of the present invention. Effect of the Invention

[0070] Compared with the prior art, the beneficial effects of the present invention are as follows: The cordycepin derivatives and pharmaceutical compositions thereof according to the present invention have excellent antitumor proliferation effects. Compared with the parent drug, the cordycepin derivatives have excellent affinity to cell membranes, so that the half-life of the drug metabolized in the body is longer, and the time it remains in the body is longer. Compared with other nucleoside antitumor drugs, the cordycepin derivatives and pharmaceutical compositions thereof according to the present invention have a wider range of tumor types and effects, and have excellent inhibitory effects on gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, colorectal cancer, melanoma, ovarian cancer, etc., with reduced side effects and excellent therapeutic effects. [Brief description of the drawings]

[0071] [Figure 1] FIG. 1 shows the antitumor effects of the blank group, cordycepin control group, and compound group against a mouse model transplanted with liver cancer cells, Hep-1-6. [Diagram 2] FIG. 1 shows the inhibitory effects of a control group and each compound group on a zebrafish tumor-bearing model of small cell lung cancer H446 cells. [Diagram 3] FIG. 1 shows the suppressive effects of compound 16 and an immune checkpoint inhibitor on a mouse model implanted with colon cancer MC38 cells. [Figure 4] FIG. 1 shows the antitumor effects of a control group and a compound group on a melanoma B16-F10 xenograft mouse model. [Diagram 5] FIG. 1 shows the inhibitory effects of compound 24 and immune checkpoint inhibitors on mouse ID8 ovarian cancer allograft tumor model. [Figure 6] FIG. 1 shows the antitumor effects of a control group and a compound group on a mouse model implanted with gastric cancer BGC-823 cells. [Figure 7] FIG. 1 shows the antitumor effects of a control group and a compound group on a mouse model transplanted with pancreatic cancer Pan02-luc cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0072] The experimental methods described in the following examples are all conventional methods unless otherwise specified, the reagents and materials are all commercially available unless otherwise specified, and the drug evaluation experiments, including cell models and animal models, are all commercially available unless otherwise specified.

[0073] Cordycepin described in the present invention is obtained from the preparation of bio-fermentation route (China Patent No. 111117896 specification), the preparation method of cordycepin or its derivatives is prepared according to the method of the specification, the post-treatment method mainly adopts the operation mode of the post-treatment of traditional organic experiments such as filtration, quenching, extraction, rotary evaporation, recrystallization, column chromatography, etc., but is not limited thereto. The preparation method of the compounds described in the examples is not for all compounds, and for the convenience of explanation, only the preparation process of the representative compounds is listed in this specification.

[0074] The cell and animal model experiments related to the anti-cancer properties of cordycepin or its derivatives in the examples are not all of the effective experimental results, but are merely relevant experiments cited to explain the effects, and drug evaluation experiments including cell and animal models are in accordance with ethical regulations.

[0075] Regarding the rearing of mouse models in drug evaluation, mice are reared in laminar flow cabinets, five nude mice per cage are reared, water and feed are added once every three days, bedding is changed once a week, the number of mice reared in each cage is no more than five, and mice in a single cage are not reared as much as possible, and mice in a single cage are kept as close together as possible; animal care must comply with the national standards for animal care and management, and attention must be paid to the behavior of animals to avoid the occurrence of unnecessary stress, and normal physiological and behavioral needs of animals such as defecation, urination, maintaining a constant body temperature, normal activity, posture adjustment, and reproduction must be satisfied; the ventilation is good, the animals are kept dry, water and feed can be freely consumed, and the operation of replenishing, replacing, and cleaning is easy; a strong and safe environment must be provided to avoid accidents such as animal escape and limbs getting caught in the gaps; sharp edges or protruding objects must be avoided to cause harm to animals; and the animals must be observed not to be disturbed.

[0076] The zebrafish model was wild-type AB strain zebrafish from the School of Biological and Pharmaceutical Engineering, Nanjing University of Technology, and was bred by natural mating. There were 30 fish in each experimental group, aged 2 dpf. They were reared in fish farming water at 28°C (water quality: 200 mg of fast-dissolving sea salt per L of reverse osmosis water, conductivity 480-510 μS / cm, pH 6.9-7.2, hardness 53.7-71.6 mg / L CaCO3), and the rearing management met the requirements of the international AAALAC certification.

[0077] The tumor cell proliferation inhibitory activity of a compound was measured by the MTT method. A single cell in good logarithmic growth phase was taken, digested with 0.25% trypsin to remove adherent cells, and then diluted with 2–4 × 10 4 The cells / mL were counted to obtain a cell suspension. The cell suspension was inoculated into a 96-well plate at 90 μL / well and cultured in a constant temperature CO2 incubator for 24 hours. The obtained compounds were added for testing and cultured at 10 μL / well for 72 hours. 10 μL / well of MTT reagent was added to the 96-well plate, and the mixture was reacted in an incubator for 4 hours, after which the supernatant was aspirated, and 100 μL / well of dimethyl sulfoxide was added. After the crystals were dissolved, the absorbance of each well was measured at a wavelength of 570 nm using a microplate reader, and the cell inhibition rate was calculated. An S-shaped curve was created for the compound concentration and the corresponding inhibition rate. The IC of the corresponding compound was calculated. 50 obtained.

[0078] Example 1 Preparation of the compound: 2-(6-amino-9H-purin-9-yl)-5-(2-hydroxyethoxy)methyl)tetrahydrofuran-3-ol

[0079] (1) Methods for protecting and deprotecting the hydroxyl group of cordycepin Method for protecting and deprotecting the hydroxyl groups at R1 and R4 positions of cordycepin 1a [ka]

[0080] Protection: 0.251 g (1 mmol) of cordycepin, 0.3618 g (2.4 mmol) of TBSCl, and 0.3404 g (5 mmol) of imidazole were added to 10 mL of DMF, and the mixture was stirred at room temperature for 10 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, and the organic phase was concentrated to give 1a (0.4607 g, yield 96.03%, MSI-MS: 480.8 [M+H] + ) was obtained.

[0081] Deprotection: 10 ml of a solvent (tetrabutylammonium fluoride:THF=2:1) ​​was prepared, 0.48 g (1 mmol) of 1a was added, and the mixture was stirred at room temperature for 5 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, and the organic phase was concentrated and recrystallized to obtain cordycepin (0.246 g, yield 97.96%).

[0082] Methods for protection 1b and deprotection of the hydroxyl group at R1 position of cordycepin [ka]

[0083] Protection: 0.251 g (1 mmol) of cordycepin, 0.3618 g (2.4 mmol) of TBSCl, and 0.3404 g (5 mmol) of imidazole were added to 10 mL of DMF, and the mixture was stirred at room temperature for 10 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, and the organic phase was concentrated to obtain 0.4607 g of 1a (yield 96.03%). Furthermore, 10 mL of solvent (acetic acid: water: THF = 13: 7: 3) was prepared, 0.48 g (1 mmol) of 1a was added, and the mixture was stirred at room temperature for 15 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, and the organic phase was concentrated to obtain 1b (0.293 g, yield 79.99%, MSI-MS: 366.5 [M + H]) by column chromatography. + ) was obtained.

[0084] Deprotection: 0.366 g (1 mmol) of 1b was added to 10 mL of 0.1 M hydrochloric acid in methanol, and the mixture was stirred at room temperature for 10 hours. After the reaction was completed, the pH was adjusted to neutral using 0.1 M aqueous sodium carbonate solution, and the mixture was extracted with ethyl acetate. The organic phase was concentrated, and then rotary evaporated or recrystallized to obtain cordycepin (0.215 g, yield 85.56%).

[0085] Methods for protection 1c and deprotection of the hydroxyl group at R4 position of cordycepin [ka]

[0086] Protection: In a nitrogen atmosphere, 1.256 g (5 mmol) of cordycepin was added to 10 mL of dichloromethane and 10 mL of diisopropylethylamine solvent, followed by dropwise addition of 0.451 g (1.6 mmol) of TBDPSCl. The mixture was stirred at room temperature for 2 hours, concentrated, and then purified by column chromatography to obtain 1c (2.20 g, yield 89.9%, MSI-MS: 366.5 [M+H] + ) was obtained.

[0087] Deprotection: 10 ml of a solvent (tetrabutylammonium fluoride:THF=2:1) ​​was prepared, 0.490 g (1 mmol) of 1c was added, and the mixture was stirred at room temperature for 5 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, and the organic phase was concentrated and recrystallized to obtain cordycepin (0.244 g, yield 97.01%).

[0088] (2) Specific method for preparing the compound 2-(6-amino-9H-purin-9-yl)-5-(2-hydroxyethoxy)methyl)tetrahydrofuran-3-ol Using compound 1b as the raw material, 3.65g (10mmol) of 1b and 0.81g (10mmol) of compound 2-chloro-ethanol were added to a round-bottom flask, 60mL of methyl ethyl ketone was added to the round-bottom flask to heat and dissolve the reactant, and 2.07g (15mmol) of potassium carbonate was added to the round-bottom flask. The reaction was carried out for 8 hours in an environment of 80°C, and the progress of the reaction was monitored by TLC. After the reaction was completed, it was filtered, the reaction solution was concentrated, and extracted twice by adding 60mL of water and 60mL of ethyl acetate. The organic phases were combined and evaporated under reduced pressure to obtain a viscous liquid. The viscous liquid was deprotected, and compound 1 (weight 2.24g, yield 76%) was obtained by column chromatography. The detection result of the obtained compound 1 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.56(s,1H),8.35(s,1H),7.09(s,2H),6.14(d,1H),5.47-5.3(d,2H), 5.01(m,1H),4.12(m,1H),3.74(m,1H),3.61-3.56(m,4H),3.51-3.46(m,2H),2.04-1.92(m,2H). 13 CNMR(100MHz,DMSO-d6)δ156.1,152.4,149.8,140.1,119.6,95.2,82.1,75.2,74.7,70.4,61.1,34.5.MSI-MS:296.3[M+H] + It was.

[0089] Example 2 Preparation of the compound: (5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl) methyl dihydrogen phosphate According to the protection method of Example 1, 3.65g (10mmol) of compound 1b was taken, and 150mL of triethyl phosphite and 4.62g (30mmol) of phosphorus oxychloride were added under ice bath conditions. The reaction was carried out under 0°C environment for 2 hours, and after the reaction was completed, 1000mL of water was added under ice bath environment to quench the reaction, and dichloromethane was added for multiple extractions, and the organic phases were combined, and the solvent was removed by rotary evaporation to obtain a viscous liquid. The obtained viscous liquid was subjected to liquid phase separation purification (C18 preparation column, Waters liquid phase preparation, mobile phase: 30% acetonitrile aqueous solution, flow rate: 2mL / min), and the effluent of the peak of the sample was concentrated, and then the phosphorylated compound (total weight 3.02g, yield 70%) was obtained.

[0090] Compound 2 was obtained according to the deprotection procedure of 1b in Example 1. The detection result of the obtained compound 2 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.54(s,1H),8.36(s,1H),7.11(s,2H),6.19(d,1H),5.15( d,1H),4.28-4.23(m,2H),4.2(s,2H),4.02(m,1H),3.71(m,1H),2.06-1.90(m,2H). 13 CNMR(100MHz,DMSO-d6)δ156.2,152.7,149.5,140.2,119.3,97.2,76.1,74.6,69.5,34.5.MSI-MS:332.2[M+H] + It was.

[0091] Example 3 (((((5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid [ka]

[0092] 36.5g (100mmol) of compound 1b was added to a 500mL reaction flask, and 150mL of DMF was added as a solvent, followed by heating and dissolving. After quenching to room temperature, 6.4g (267mmol) of 80% NaH was added in portions while stirring, and the mixture was stirred at room temperature for 15 minutes. The reaction solution was cooled to -10°C in a low-temperature reactor, and 45.1g (140mmol) of p-toluenesulfonyloxymethylphosphonic acid diethyl was added dropwise. After the dropwise addition was completed, the mixture was stirred and reacted at this temperature for 1 hour, gradually raised to room temperature and stirred for 4 hours. After the reaction was completed, glacial acetic acid was added dropwise to adjust the pH to neutral, the reaction solution was filtered, washed with dichloromethane, the filtrate was extracted three times with water (60mL), the dichloromethane phase was combined, the solvent was distilled off under reduced pressure, and toluene was added to the residue to recrystallize. The crystal mother liquor was filtered, and the filter cake was dried under reduced pressure at 50 °C to obtain a white powdery solid 3a (weight 26.87 g, yield 52%, MSI-MS: 540.6 [M+23] + ) was obtained.

[0093] Compound 3a (5.17 g, 10 mmol) was taken and added to a 100 mL reaction flask, n-butyronitrile (20 mL) was added, and trimethylchlorosilane (6.52 g, 60 mmol) was added dropwise at room temperature. After the dropwise addition, the reaction solution was heated and refluxed for 24 hours. After the reaction was completed, the solvent was distilled under reduced pressure and concentrated to dryness, and 20 mL of water was added to the residue. 2 M sodium hydroxide was added to adjust the pH to 8, and the mixture was extracted three times with ethyl acetate (50 mL). The aqueous phase was neutralized with 1 M hydrochloric acid until the pH was 3 to 4. The mixture was then heated to 70 to 80 ° C. to crystallize, and compound 3b (weight 3.40 g, yield 74%, MSI-MS: 460.6 [M + H] + ) was obtained.

[0094] According to the deprotection procedure of Example 1b, compound 3b is treated to obtain compound 3, and the detection result of the prepared compound is: 1 HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.32(s,1H),7.13(s,2H),6.12(d,1H),5.39(d,1H), 4.81(s,2H),4.02(m,1H),3.98(m,1H),3.74(d,2H),3.63-3.56(m,2H),2.08-1.94(m,2H).13 CNMR(100MHz,DMSO-d6)δ156.7,152.6,149.1,140.4,119.5,98.2,81.1,76.3,74.3,73.1,34.7.MSI-MS:346.3[M+H] + It was.

[0095] Example 4 Preparation of the compound: (5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methyl diisobutyl phosphate [ka]

[0096] 3.65g (10mmol) of compound 1b was added to a 500mL reaction flask and dissolved in 100mL of anhydrous DMF, and 4.96g (15mmol) of diisobutyl (4-nitrophenyl) phosphate was added. Furthermore, 1.17g (10mmol) of tert-butyl magnesium chloride was dissolved in 20mL of THF, and the solution was gradually added dropwise to the above reaction solution. The reaction solution was gradually warmed to room temperature and reacted for 2 hours, and the reaction was monitored on a TLC plate. After the reaction was completed, the reacted mixture was left to stand, diluted with 100mL of ethyl acetate, washed and extracted three times with 50mL of saturated aqueous sodium bicarbonate solution, and then extracted with 50mL of saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained oil was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10:1) to obtain compound 4a (weight 4.24g, yield 76%).

[0097] According to the deprotection procedure of 1b in Example 1, 4a was treated to obtain compound 4. The detection result of the prepared compound 4 was: 1HNMR(400MHz,DMSO-d6)δ8.52(s,1H),8.31(s,1H),7.11(s,2H),6.15(d,1H),5.33(d,1H),4.24-4 .05(m,2H),4.02(m,1H),3.93(m,4H),3.77(m,1H),2.08-1.83(m,2H),1.33(m,2H),0.90(d,12H). 13 CNMR(100MHz,DMSO-d6)δ154.3,151.7,148.4,141.9,119.2,98.2,74.1,74.0,73.5,68.1,34.2,28.6,19.5.MSI-MS:466.7[M+Na] + It was.

[0098] Example 5 Preparation of the compound: ((((((((5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphoryl)bis(oxy)bis(methylene)bis(2-methylpropionic acid) [ka]

[0099] Compound 5 was prepared according to the preparation method of 3a in Example 3, in combination with the deprotection method of 1b in Example 1, in which diethyl p-toluenesulfonyloxymethylphosphonate was replaced with an equivalent amount of (((tolyloxy)methyl)phosphoryl)bis(oxy))bis(methylene)bis(2-methylpropionic acid), to obtain compound 5 (total yield 74%). The detection result of the obtained compound 5 was: 1 HNMR(400MHz,DMSO-d6)δ8.57(s,1H),8.33(s,1H),7.11(s,2H),6.84(d,4H),6.18(d,1H),5.46(d,1H), 4.12(m,1H),3.94(m,1H),3.81(d,2H),3.60-3.44(m,2H),2.55(m,2H),2.08-1.82(m,2H),1.14(d,12H). 13CNMR(100MHz,DMSO-d6)δ174.2,155.7,153.6,149.5,141.2,119.5,99.4,93.2,77.4,74.5,74.1,71.5,34.2,33.6,19.3.MSI-MS:546.5[M+H] + It was.

[0100] Example 6 Preparation of compound: methyl((5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-D-valine. The preparation steps are as follows: [ka]

[0101] The method for preparing methyl((4-nitrophenoxy)(phenoxy)phosphoryl)-D-valine is as follows.

[0102] 1.17 g (10 mmol) of valine was dissolved in 50 mL of dichloromethane. The solution was cooled to about 0° C., and 2.11 g (10 mmol) of phenyl dichlorophosphate was added, followed by slowly dripping in 2.02 g (20 mmol) of triethylamine, and the reaction was warmed to room temperature and stirred for 80 minutes. 1.39 g (10 mmol) of the reaction product p-nitrophenol was added, followed by dripping in 2.02 g (20 mmol) of triethylamine, and stirred at room temperature for 180 minutes. After the reaction was completed, the reaction was washed with diethyl ether, and the resulting solid was removed by filtration. The filtrate was concentrated with a rotary evaporator, and the obtained sample was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate (1:1)), and the detection results of the obtained compound were as follows: 1 HNMR(400MHz,DMSO-d6)δ8.25(d,2H),7.40(m,2H),7.35(d,2H),7.20(m,3H),3.68(s,1H),3.65(s,3H),3.35(m,1H),1.90(m,2H),0.86(m,3H). 13CNMR(100MHz,DMSO-d6)δ171.6,156.5,150.5,141.1,130.2,126.5,121.7,121.4,120.4,52.2,51.9,26.4,9.6.MSI-MS:395.3[M+H] + It was.

[0103] According to the preparation method of 4a in Example 4, in combination with the deprotection method of 1b in Example 1, diisobutyl(4-nitrophenyl)phosphate was replaced with an equivalent amount of methyl((4-nitrophenoxy)(phenoxy)phosphoryl)-D-valine to obtain compound 6 (yield 68%). The detection result of the finally obtained compound 6 was: 1 HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.33(s,1H),7.43(m,2H),7.20(m,3H)7.08(s,2H),6.15(d,1H),5.47(d,1H),4.26-4 .12(m,2H),4.02(m,1H),3.75(m,1H),3.67(d,1H),3.62(s,3H),3.28(d,1H),2.41(m,1H),2.08-1.96(m,2H),0.98(d,6H). 13 CNMR(100MHz,DMSO-d6)δ173.6,154.9,151.5,150.6,149.8,141.3,132.7,123.3,12 0.8,119.2,98.2,76.1,74.5,72.0,57.6,52.3,35.2,32.4,20.1.MSI-MS:521.4[M+H] + It was.

[0104] Example 7 Preparation of compound: isopropyl(((((((((5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)(phenoxy)phosphoryl)alanine ester, the preparation steps are as follows: [ka]

[0105] The method for preparing isopropyl(phenoxy((tolyloxy)methyl)phosphoryl)alanine ester is as follows.

[0106] In a 1000mL reaction flask, 216.15g (1.65mol) of alanine isopropyl ester, 155g (1.65mol) of phenol, and 240mL of chloroform were added, and 138.8g (1mol) of phosphorus trichloride was added dropwise from around 5℃ while stirring under ice-salt bath conditions, and the reaction temperature was controlled between 5-10℃. After the dropwise addition of phosphorus trichloride was completed, stirring was continued for 30 minutes, the ice-salt bath was removed, the temperature was gradually raised to room temperature, and stirring was continued for 2 hours, after which the pressure was reduced at 50℃ to remove hydrogen chloride gas. The reaction product was cooled to room temperature, and 80mL of saturated aqueous sodium bicarbonate solution was slowly added, followed by the addition of solid sodium bicarbonate powder to adjust the pH of the reaction solution to 7-8. The precipitated salt was removed by filtration, the filtrate was placed in a separating funnel to separate the chloroform layer, the aqueous layer was extracted once with 100 mL of chloroform, the organic phases were combined and washed once each with a saturated aqueous sodium bicarbonate solution and water, the chloroform layer was taken and distilled under reduced pressure, the residue was the target product, phenylalanine isopropyl chlorophosphate, with a yield of 87%.

[0107] In a 1000mL reaction flask, 306g (1mol) of phenylalanine isopropyl chlorophosphate, 40.6g (1.35mol) of paraformaldehyde, 14mL of triethylamine, and 260mL of toluene were added. The reaction was gradually heated to 105°C in a nitrogen atmosphere, and the reaction was vigorously carried out at reflux for 3 hours. After the reaction was completed, the ice-salt bath was cooled to 0°C, 169.5g (0.9mol) of p-toluenesulfonyl chloride was added, 220mL of toluene was added, and 177mL of triethylamine was added dropwise. The reaction was stirred for 2 hours while maintaining the reaction temperature at 0°C, and the temperature was gradually raised to room temperature and stirred for 12 hours. After the reaction was completed, the mixture was filtered, the filter cake was washed with toluene (50mL x 3), and the mother liquor was combined and washed with 5% aqueous sodium carbonate solution (200mL x 2) and water (200mL x 2) in sequence, and the layers were separated to obtain an organic phase. The organic phase was distilled under reduced pressure to obtain a pale yellow oily product, isopropyl(phenoxy((tolyloxy)methyl)phosphoryl)alanine ester, which was used in the subsequent reaction. The weight was 291 g, and the yield was 64%. The detection result of the obtained compound was: 1 HNMR(400MHz,DMSO-d6)δ7.74(d,2H),7.45-7.39(m,4H),7.22(m,3H),4.95(m,1H) ),3.91(d,2H),3.65(s,1H),3.57(m,1H),2.42(s,3H),1.28(d,3H),1.18(d,6H). 13 CNMR(100MHz,DMSO-d6)δ171.6,150.4,144.4,140.3,130.5,130.1,128.5,121.4,120.2,69.6,63.8,50.5,21.7,21.5,19.2.MSI-MS:456.5[M+H] + It was.

[0108] According to the preparation method of 3a in Example 3, in combination with the deprotection method of 1b in Example 1, compound 7 was prepared, in which p-toluenesulfonyloxymethylphosphonic acid diethyl was replaced with isopropyl(phenoxy((tolyloxy)methyl)phosphoryl)alanine ester in an equivalent amount to obtain compound 7 (yield 52%). The detection result of the obtained compound 7 was: 1HNMR(400MHz,DMSO-d6)δ8.59(s,1H),8.37(s,1H),7.45(m,2H),7.24(m,3H),7.12(s,2H),6.16(d,1H),5.33(d,1H),4. 98(m,1H),4.08-3.99(m,2H),3.84(m,1H),3.77(m,2H),3.58-3.42(m,3H),2.08-1.96(m,2H),1.29(d,3H),1.16(d,6H). 13 CNMR(100MHz,DMSO-d6)δ172.1,156.3,152.5,150.4,149.8,140.1,130.2,122.1,120.5 ,119.1,98.2,79.1,74.8,74.7,73.1,69.6,52.3,34.5,22.5,18.8.MSI-MS:535.6[M+H] + It was.

[0109] Example 8 2-(-5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-3-hydroxypropyl valine salt [ka]

[0110] Using compound 1b as a raw material, 3.65 g (10 mmol) of 1b and 2.10 g (10 mmol) of compound 2-chloro-3-hydroxypropylvaline were added to a round-bottom flask, 60 mL of methyl ethyl ketone was added to the round-bottom flask to heat and dissolve the reactant, and 2.07 g (15 mmol) of potassium carbonate was added to the round-bottom flask. The reaction was carried out for 10 hours in an environment of 80 ° C., and the progress of the reaction was monitored by TLC. After the reaction was completed, it was filtered, the reaction solution was concentrated, and extracted twice by adding 60 mL of water and 60 mL of ethyl acetate. The organic phases were combined and evaporated under reduced pressure to obtain a viscous liquid. The viscous liquid was subjected to column chromatography to obtain compound 8a (weight 3.24 g, yield 52%, MSI-MS: 539.6 [M + H] + ) was obtained.

[0111] Compound 8a was used as a substrate, and compound 8 (yield 91%) was obtained by the deprotection method of 1b in Example 1. The detection results of the obtained compound 8 were as follows: 1 HNMR(400MHz,DMSO-d6)δ8.88(s,2H),8.54(s,1H),8.37(s,1H),7.14(s,2H),6.18(d,1H),5.57(d,1H),4.33 -4.11(m,3H)4.03(m,1H),3.96-3.89(m,3H),3.61-3.36(m,4H),2.37(m,1H),2.08-1.82(m,2H),0.98(d,6H). 13 CNMR(100MHz,DMSO-d6)δ173.5,154.6,153.5,150.4,141.3,119.6,99.5,85 .6,75.7,74.0,73.1,63.3,62.5,58.7,34.7,30.8,18.8.MSI-MS:425.4[M+H] + It was.

[0112] Example 9 Preparation of the compound (4-amino-6-(6-amino-9H-purin-9-yl)-2,2-dioxide-1,7-dioxa-2-azaspiro[4.4]non-3-en-8-yl) methyl dihydrogen phosphate, the preparation method is as follows. [ka]

[0113] Compound 2 was used as the raw material to react, and 9.00 g (27.2 mmol) of compound 2 and 4.88 g (27.2 mmol) of CbzCl were added to 38.5 mL of toluene and 38.5 mL of water, and 4.70 g (34 mmol) of K2CO3 was added, and the mixture was vigorously stirred at a temperature below 25 °C. After stirring at room temperature for 3 hours, 0.275 g (2.72 mmol) of triethylamine and 5.78 g of sodium chloride were added sequentially, and the mixture was stirred for another 30 minutes. The organic layer was separated and concentrated to give the desired oily product 9a (weight 11.38 g, yield 90%, MSI-MS: 488.7 [M+Na] + ) was obtained.

[0114] 9.3 g (20 mmol) of compound 9a was dissolved in 60 mL of acetone by stirring at room temperature, and 5.50 mL of Jones reagent (2.2 M dichromium trioxide, 12.1 mmol) was added to the solution and added dropwise at room temperature for 2 h. The resulting reaction mixture was stirred at room temperature for another hour, filtered, and concentrated under reduced pressure to obtain an oily liquid. This liquid was dissolved in 30 mL of diethyl ether, washed once with 30 mL of saturated ammonium chloride solution, dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and then subjected to column chromatography to obtain compound 9b (weight 3.98 g, yield 43%, MSI-MS: 464.4 [M+H] + ) was obtained.

[0115] 4 mL (30 mmol) of trimethylsilyl cyanide and 2.53 mL (20 mmol) of boron trifluoride diethyl ether were added to a solution of 9.26 g (20 mmol) of 9b in 50 mL of dichloromethane. The mixture was stirred at room temperature for 2 h and the solvent was evaporated to dryness. The resulting residue was dissolved in 100 mL of ethyl acetate, washed twice with 50 mL of brine and dried (Na2SO4). After filtration and evaporation of the solvent, it was purified by column chromatography (n-hexane / ethyl acetate, 1:2) to give the white compound 9c (weight 7.45 g, yield 76%, MSI-MS: 491.9 [M+H] + ) was obtained.

[0116] 1.9 mL of Et3N (14 mmol) was added to 5 mL of anhydrous dichloromethane, and 1.08 g (2.2 mmol) of compound 9c was added to the solution to dissolve. After the mixture was cooled to -30°C, 460 μL (6 mmol) of methanesulfonyl chloride was slowly added. The mixture was kept stirring at -20°C for 1 h and at 0°C for 1 h. The volatiles were then removed by concentration under reduced pressure, and the residue was dissolved in 10 mL of ethyl acetate, followed by washing twice successively with 10 mL of water and 10 mL of brine. The organic phase (Na2SO4) was dried, concentrated under reduced pressure, and purified by column chromatography (n-hexane:ethyl acetate, 10:1) to obtain a white amorphous solid 9d (weight 0.975 g, yield 78%). The detection results of the preparation were 1HNMR(400MHz,DMSO-d6)δ10.45(s,1H),8.36(s,1H),8.20(s,1H),7.34-7.32(s,5H),6. 12(s,1H),4.65(s,2H),4.30-4.03(m,4H),3.74(m,1H),3.15(s,3H),2.41-2.16(m,2H). 13 CNMR(100MHz,DMSO-d6)δ153.6,152.5,151.7,149.8,140.2,136.4,128.9,127.7, 127.2,123.5,120.4,97.4,71.6,69.2,68.2,66.9,38.2,37.8.MSI-MS:569.4[M+H] + It was.

[0117] 490 mg (1.5 mmol) of cesium carbonate was added to a suspension of 0.568 g (1 mmol) of 9d in 3 mL of anhydrous acetonitrile, and the mixture was stirred at room temperature for 2 h. The solvent was removed and the resulting residue was dissolved in 20 mL of ethyl acetate and washed twice with 10 mL of water and 10 mL of brine in sequence. The organic phase (Na2SO4) was dried, filtered, and then concentrated under reduced pressure. Purification by column chromatography (n-hexane / ethyl acetate, 3:1) gave a white solid 9e (weight 4.49 g, yield 79%, MSI-MS: 569.4 [M+H]). + ) was obtained.

[0118] 5.68 g of compound 9e (10 mmol) was dissolved in 200 mL of methanol. Then, 1.5 g of ammonium formate (30 mmol) and 0.75 g of 10% Pd-C were added, and the reaction mixture was stirred at room temperature for 10 minutes and then heated to reflux for 45 minutes. The mixture was filtered through diatomaceous earth, and the filtrate was evaporated to dryness to obtain compound 9 (4.12 g, 95% yield). The detection result of the obtained compound 9 was 1 HNMR(400MHz,DMSO-d6)δ8.59(s,1H),8.38(s,1H),7.14(s,2H),6.88(s,2H),6.16(d,1) H),5.25(s,1H),4.28(m,1H),4.18(s,2H),4.02(m,1H),3.74(m,1H),2.09-1.96(m,2H). 13CNMR(100MHz,DMSO-d6)δ168.5,155.1,151.4,147.8,141.1,119.4,98.2,88.9,86.1,73.0,68.1,37.9.MSI-MS:435.3[M+H] + It was.

[0119] Example 10 Preparation of the compound: N-(3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)octadecamide [ka]

[0120] 2.51 g (10 mmol) of cordycepin was taken, and 40.00 mL of anhydrous pyridine and 8.5 mL of acetic anhydride were added under ice bath conditions. The reaction was monitored by HPLC liquid phase, and the reaction was terminated after about 5 hours. The solvent was removed to give a viscous liquid 10a (weight 2.68 g, yield 80%, MSI-MS: 358.6 [M+Na] + ) was obtained.

[0121] Compound 10a (3.35 g, 10 mmol) and octadecanoic acid chloride (2.89 g, 10 mmol) were taken, and 60 mL of anhydrous pyridine was added in an ice bath environment. The temperature was gradually raised to 40° C. and the mixture was reacted for 10 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, water and ethyl acetate were added for extraction, and the organic phase was back-extracted. The organic phase was recovered and rotary evaporated to obtain an oily liquid 10b. The purified product 10b was obtained by column chromatography (5.40 g, yield 90%, MSI-MS: 624.4 [M+Na] + ) was obtained.

[0122] Compound 10b (6.01 g, 10 mmol) was taken, dissolved in 450 mL of methanolic ammonia solution, reacted, stirred at room temperature, and the reaction was detected by thin layer chromatography. The reaction was stopped after 4 hours. The solvent was removed to obtain the target product 10 (weight 4.67 g, yield 90%).

[0123] Alternatively, the protection and deprotection were performed by referring to the preparation method of 1a in Example 1, and the protected 1a was subjected to an acylation reaction in this second step, followed by deprotection to obtain compound 10. The detection result of the obtained compound 10 was as follows: 1 HNMR(400MHz,DMSO-d6)δ10.55(s,1H),8.64(s,1H),8.38(s,1H),6.18(d,1H),5.35(d,1H),5.04(m,1H),4.02(m,1H), 3.76(m,1H),3.58(m,1H),3.52(m,1H),2.35(m,2H),2.07-1.94(m,2H),1.58(m,2H),1.30-1.26(m,28H),0.89(m,3H). 13 CNMR (100MHz, DMSO-d6) δ 173.5, 153.3, 152.5, 148.8, 141.0, 122.6, 99.2, 84.1, 74.3, 63.8, 38.5, 34.8, 31.5, 29.5, 28.8, 25.6, 22.4, 14.7. (Partial alkyl peak overlap) MSI-MS: 540.7 [M+Na] + It was.

[0124] Example 11 N-(3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)cyclopentanecarboxamide According to the method of Example 10, the steps of protecting, acylating, and deprotecting cordycepin were carried out to obtain compound 11 (total yield 72%), and octadecanoic acid chloride was replaced with cyclopentanecarbonyl chloride. The detection result of the obtained compound 11 was as follows: 1 HNMR(400MHz,DMSO-d6)δ10.52(s,1H),8.66(s,1H),8.38(s,1H),6.16(d,1H),5.28(d,1H),5.01(m, 1H),4.01(m,1H),3.77(m,1H),3.57-3.46(m,2H),2.46(m,1H),2.08-1.88(m,2H),1.80-1.55(m,8H). 13CNMR(100MHz,DMSO-d6)δ172.5,153.3,151.5,149.5,139.8,123.6,98.4,82.1,74.9,63.8,49.5,34.5,32.4,24.5.MSI-MS:348.2[M+H] + It was.

[0125] Example 12 N-(3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)isonicotinamide According to the method of Example 10, cordycepin was protected, acylated, and deprotected to obtain compound 12 (total yield 76%), and octadecanoic acid chloride was replaced with pyridine-3-carboxylic acid chloride. The detection result of the obtained compound 12 was as follows: 1 HNMR(400MHz,DMSO-d6)δ11.05(s,1H),8.86(d,2H),8.41(s,1H),8.22(s,1H),8.01(d,2H),6.15( d,1H),5.22(d,1H),4.98(m,1H),4.02(m,1H),3.75(m,1H),3.58-3.46(m,2H),2.08-1.88(m,2H). 13 CNMR(100MHz,DMSO-d6)δ165.5,152.3,151.9,149.8,149.7,140.9,140.4,123.4,120.5,99.4,82.8,74.7,63.8,35.4.MSI-MS:357.3[M+H] + It was.

[0126] Example 13 (5-(6-amino-2-fluoro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl) dihydrogen methyl phosphate [ka]

[0127] 193 μL (1.39 mmol) of 2,2,2-trifluoroacetic anhydride was added to a solution of 428 mg (1.40 mmol) of tetrabutylammonium nitrate in dry dichloromethane (15 mL) at 0° C. to obtain the nitration mixture. After reacting for 45 min at 0° C., the solution was slowly added to 480 mg (1 mmol) of compound 1a in dry dichloromethane (15 mL). After reacting for 14 h at room temperature in the dark, the reaction mixture was poured into a cold mixture of H2O (50 mL), saturated NaHCO3 (35 mL) and CHCl2:Et2O (1:2, 30 mL) and extracted twice with CHCl2:Et2O (1:2, 30 mL). The organic extract was washed with brine, dried over anhydrous Na2SO4, and evaporated under vacuum (temperature was kept below 40° C.). The crude product was purified by column chromatography, eluted with CH2Cl2 and then purified with CH2Cl2:acetone (99:1 to 95:5) to give compound 13a (weight 273 mg, yield 52%, MSI-MS: 525.4 [M+H] + ) was obtained.

[0128] To a suspension of 236 mg (0.45 mmol) of 13a in dry acetonitrile (15 mL) was added TBAF (1.3 equiv., 600 μL, 0.6 mmol) dropwise over 1 min at 0 °C. The mixture was stirred for 20 min and the resulting solution was evaporated under vacuum without heating. The crude product was purified by column chromatography (CHCl:acetone (100:0 to 90:10) to give compound 13b (wt 78 mg, yield 35%, MSI-MS: 498.8 [M+H] + ) was obtained.

[0129] The obtained compound 13b was prepared according to the phosphorylation method of Example 2 and the deprotection method of Example 1 to obtain compound 13 (yield 65%). The detection result of the obtained compound 13 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.38(s,1H),6.98(s,2H),6.16(d,1H),5.36(d,1H),4.2 8(m,1H),4.21(s,2H),4.02(m,1H),3.95(m,1H),3.74(m,1H),2.02-1.82(m,2H). 13CNMR(100MHz,DMSO-d6)δ158.3,157.2,148.8,141.5,118.6,97.2,74.9,74.6,67.1,35.5.MSI-MS:372.2[M+Na] + It was.

[0130] Example 14 (5-(6-amino-2-mercapto-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl) dihydrogen methyl phosphate [ka]

[0131] 4.80 g (10 mmol) of compound 1a was added to 60 mL of acetic acid and stirred at 40° C. until dissolved. After the solution was cooled to room temperature, 5 mL (25 mmol) of 30% hydrogen peroxide was added. The solution was stirred at 40° C. for 3 days, filtered, and the solid was recrystallized in water to give compound 14a (weight 3.37 g, yield 68%, MSI-MS: 497.4 [M+H] + ) was obtained.

[0132] 4.96 g (10 mmol) of 14a was added to 40 mL of 3M aqueous hydrochloric acid, and the mixture was heated under reflux until the entire solid was dissolved. After refluxing for 10 minutes, the mixture was cooled to room temperature, the solvent was removed, and 20 mL of ethanol was added. The precipitated crystals were filtered and dried to obtain compound 14b (weight 2.92 g, yield 62%, MSI-MS: 487.5 [M+H] + ) was obtained.

[0133] Compound 14b (4.86 g, 10 mmol) was dissolved in 100 mL of water, and 1.0 g of activated Raney Ni was added in portions, followed by introduction of hydrogen gas. The reaction mixture was stirred at 55° C. for 4 days. The introduction of hydrogen gas was stopped, inorganic salts were removed by filtration, the solvent was removed by distillation under reduced pressure, and the crude product was washed with ethanol and diethyl ether, respectively, and dried to obtain a gray solid 14c (weight 3.30 g, yield 70%, MSI-MS: 495.8 [M+Na] + ) was obtained.

[0134] Methanol, pyridine and carbon disulfide were mixed in a volume ratio of 4:5:2 to obtain 50 mL of a solution, to which 4.72 g (10 mmol) of 14c was added, and a balloon was placed on the reflux condenser to prevent carbon disulfide from volatilizing. The reaction mixture was stirred at 40°C for 4 days. After filtration, the crude product was heated to reflux in a 5% mass fraction aqueous sulfuric acid solution for 20 minutes, and filtered while heating to remove impurities. The mixture was cooled to room temperature, filtered and dried to obtain gray crystals 14d (weight 2.82 g, yield 55%). The obtained compound 14d was prepared according to the phosphorylation method of Example 2 and the deprotection method of Example 1 to obtain compound 14 (yield 71%). The detection result of the obtained compound 14 was as follows: 1 HNMR(400MHz,DMSO-d6)δ12.22(s,1H),8.36(s,1H),6.99(s,2H),6.16(d,1H),5.33(d,1) H),4.30(m,1H),4.21(s,2H),4.02(m,1H),3.94(m,1H),3.71(m,1H),2.06-1.88(m,2H). 13 CNMR(100MHz,DMSO-d6)δ154.3,152.1,149.2,139.8,118.6,99.2,74.8,74.3,67.1,35.1.MSI-MS:364.3[M+H] + It was.

[0135] Example 15 (5-(6-amino-9H-purin-9-yl)-3-fluoro-4-hydroxytetrahydrofuran-2-yl) dihydrogen methyl phosphate [ka]

[0136] In 10 mL of DMF, 2.67 g (10 mmol) of adenosine, 3.618 g (24 mmol) of TBSCl, and 3.95 g (50 mmol) of pyridine were added and stirred at room temperature for 10 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, and the organic phase was concentrated and purified by column chromatography to obtain 15a (2.33 g, yield 47%, MSI-MS: 497.7 [M+H] + ) was obtained.

[0137] A solution of 4.96g (10mmol) of 15a and pyridine (1.5mL) in dichloromethane (100mL) was stirred under nitrogen atmosphere and cooled to -5℃, and 2.5mL (15mmol) of trifluoromethanesulfonic anhydride was added dropwise and reacted at 0℃ for 2 hours. After TLC showed the completion of the reaction, it was poured into ice water (100mL), stirred and separated, and the aqueous phase was extracted with dichloromethane (100mL). The dichloromethane phase was combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was recrystallized by adding petroleum ether (40mL), filtered, and dried to obtain 15b (weight 5.65g, yield 90%). The detection results of the preparation were as follows: 1 HNMR(400MHz,DMSO-d6)δ8.59(s,1H),8.35(s,1H),7.11(s,2H),6.15(d,1H),5.05 (m,1H),4.55(m,1H),4.12(m,1H),4.02-3.79(m,2H),0.99(s,18H),0.20(s,12H). 13 CNMR(100MHz,DMSO-d6)δ156.2,152.5,149.7,140.2,119.5,118.4,96.5,86.4,83.5,73.8,63.2,30.8,30.6,25.8,0.MSI-MS:628.8[M+H] + It was.

[0138] In a nitrogen gas atmosphere, 6.28 g (10 mmol) of 15b was dissolved in ethyl acetate (40 mL), 10 mL (22 mmol) of a 37% solution of hydrofluoric acid in triethylamine was added, and the mixture was stirred and heated to 70° C. for about 8 hours. After TLC showed the completion of the reaction, the mixture was cooled to room temperature, washed with saturated sodium bicarbonate solution until neutral, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was recrystallized with anhydrous methanol (30 mL) to obtain solid 15c (weight 2.04 g, yield 41%). The obtained compound 15c was prepared according to the phosphorylation method of Example 2 and the deprotection method of Example 1 to obtain compound 15 (yield 77%). The detection result of the obtained compound 15 was as follows: 1HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.37(s,1H),7.07(s,2H),6.15(d,1H),5.33( d,1H),4.70(m,1H),4.44(m,1H),4.28(m,1H),4.18(s,2H),4.02(m,1H)3.55(m,1H). 13 CNMR(100MHz,DMSO-d6)δ155.3,151.4,148.9,141.1,119.6,97.2,90.1,79.0,73.4,61.5.MSI-MS:350.2[M+H] + It was.

[0139] Example 16 Isopropyl((((((((5-(6-amino-9H-purin-9-yl)-4-cyanotetrahydrofuran-2-yl)methoxy)methyl)(phenoxy)phosphoryl)alanine ester [ka]

[0140] Compound 7 was obtained according to the method of Example 7, and compound 16 was prepared using compound 7 as the raw material. 5.34g (10mmol) of compound 7 was taken and dissolved in 100mL of dichloromethane solution, and 1.3mL (1.50g, 10mmol) of trifluoromethanesulfonic acid was further added. After stirring the reaction for 10 minutes, 2.7mL of trimethylsilyl triflate (10mmol) was slowly added dropwise to the solution, and the resulting mixture was stirred for 30 minutes in an environment of -40°C. Then, 3.96g (40mmol) of trimethylsilyl cyanide was slowly added, and the mixture was stirred for 2 hours. Then, 3.5mL of triethylamine was added dropwise, and the reaction mixture was heated to room temperature. Then, 5.5g of solid sodium bicarbonate and 20.7mL of water were added. The resulting mixture was stirred for 10 minutes. Then, the organic extract was obtained by extracting with dichloromethane and water, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by column chromatography to give the product compound 16 as a grey solid (weight 1.71 g, yield 32%). 1HNMR(400MHz,DMSO-d6)δ8.57(s,1H),8.36(s,1H),7.41(m,2H),7.22(m,3H),7.09(s,2H),6.17(d,1 H),4.99(d,1H),3.97(m,1H),3.74-3.33(m,6H),2.88(m,1H),2.04-1.92(m,2H),1.28-1.14(d,9H). 13 CNMR(100MHz,DMSO-d6)δ173.4,155.3,152.5,150.2,149.8,141.3,130.2,121.6,120.5 ,118.2,92.2,77.4,75.0,73.1,69.1,51.2,26.5,25.8,22.4,20.1.MSI-MS:544.5[M+H] + It was.

[0141] (Example 17) ((((((((5-(6-amino-2-fluoro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphoryl)bis(oxy)bis(methylene)bis(2,2-dimethylpropionic acid) [ka]

[0142] Compound 13b was obtained according to the method of Example 13, compound 17a was obtained according to the protection method of 1b in Example 1 using 13b as the raw material, and compound 17 was obtained according to the method of preparing 3a in Example 3 using compound 17a as the raw material. (p-toluenesulfonyloxymethyl)diethylphosphonate was replaced with (((tolyloxy)methyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionic acid) in an equivalent amount, and 1b was replaced with 17a in an equivalent amount, with the total yield being 61%. The detection result of the obtained compound 17 was as follows: 1HNMR(400MHz,DMSO-d6)δ8.33(s,1H),6.99(s,2H),6.88(s,4H),6.16(d,1H),5.37(d,1H), 4.02(m,1H),3.95(m,1H),3.85(s,2H),3.61-3.42(m,2H),2.05-1.88(m,2H),1.25(s,18H). 13 CNMR(100MHz,DMSO-d6)δ175.2,158.3,157.5,148.6,140.3,119.2,99.2,92.2,79.1,74.8,74.3,71.2,38.5,35.8,26.8.MSI-MS:592.5[M+H] + It was.

[0143] (Example 18) (((((5-(6-amino-9H-purin-9-yl)-3-fluoro-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionic acid) [ka]

[0144] Compound 18a was obtained by referring to the method of Example 15 and the protection method of 1b in Example 1, and compound 18a was used as the raw material to prepare 3a in Example 3 to obtain compound 18. (p-Toluenesulfonyloxymethyl)diethylphosphonate was replaced with (((tolyloxy)methyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionic acid) in an equivalent amount, and 1b was replaced with 18a in an equivalent amount, with a total yield of 34%. The detection result of the obtained compound 18 was: 1 HNMR(400MHz,DMSO-d6)δ8.59(s,1H),8.32(s,1H),7.09(s,2H),6.84(s,4H),6.15(d ,1H),5.35(d,1H),4.71-4.64(m,2H),3.85(m,2H),3.60-3.33(m,3H),1.27(s,18H). 13CNMR(100MHz,DMSO-d6)δ175.9,157.3,151.5,149.8,141.3,118.9,98.2,92.8,91.5,79.4,73.3,71.4,70.5,38.6,27.8.MSI-MS:592.4[M+H] + It was.

[0145] (Example 19) (((((5-(6-amino-2-mercapto-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphoryl)bis(oxy)bis(methylene)bis(2,2-dimethylpropionic acid) [ka]

[0146] Compound 14d was obtained by referring to the method of Example 14, compound 19a was obtained by using 14d as the raw material according to the protection method of 1b in Example 1, and compound 19a was obtained by using compound 19a as the raw material according to the method of preparing 3a in Example 3. (p-toluenesulfonyloxymethyl)diethylphosphonate was replaced with (((tolyloxy)methyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionic acid) in an equivalent amount, and 1b was replaced with 19a in an equivalent amount, with a total yield of 18%. The detection result of the obtained compound 19 was: 1 HNMR(400MHz,DMSO-d6)δ12.12(s,1H),8.34(s,1H),7.01(s,2H),6.84(s,4H),6.12(d,1H),5.35( d,1H),4.01(m,1H),3.93(m,1H),3.85(s,2H),3.63-3.37(m,2H),2.06-1.85(m,2H),1.27(s,18H). 13 CNMR(100MHz,DMSO-d6)δ175.8,155.4,152.5,149.7,140.5,118.9,98.7,92.8,78.1,74.9,74.7,71.2,38.7,34.5,27.5.MSI-MS:606.6[M+H] + It was.

[0147] (Example 20) Isopropyl((((5-(6-amino-2-fluoro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)(phenoxy)phosphoryl)alanine ester [ka]

[0148] Compound 17a was obtained according to the methods of Examples 13 and 17, and compound 20 was prepared by referring to the method of Example 7. (p-toluenesulfonyloxymethyl)diethylphosphonate was replaced with isopropyl(phenoxy((tolyloxy)methyl)phosphoryl)alanine ester in an equivalent amount, and 1b was replaced with 17a, with a total yield of 67%. The detection result of the obtained compound 20 was: 1 HNMR(400MHz,DMSO-d6)δ8.37(s,1H),7.40(m,2H),7.20(m,3H),7.02(s,2H),6.15(d,1H),5.38(s,1H),4. 94(m,1H),4.00(m,1H),3.95(m,1H),3.71-3.53(m,5H),3.34(m,1H),2.06-1.82(m,2H),1.28-1.14(d,9H). 13 CNMR(100MHz,DMSO-d6)δ172.1,157.3,156.8,150.0,149.8,140.2,121.3,120.6,11 9.2,97.4,76.6,75.0,74.8,72.1,69.4,50.1,34.5,21.7,19.5.MSI-MS:553.5[M+H] + It was.

[0149] Example 21 N-(2-fluoro-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)octadecamide [ka]

[0150] Compound 13b was obtained by following the method of Example 13, and compound 21a was prepared by following the method of preparing 10b from 10a in Example 10 using 13b as the raw material, and compound 21 was prepared by following the deprotection method of 1a in Example 1 using 21a as the raw material, with the total yield of 74%. The detection result of the obtained compound 21 was as follows: 1 HNMR(400MHz,DMSO-d6)δ10.53(s,1H),8.34(s,1H),6.15(d,1H),5.6(d,1H),5.02(m,1H),4.01(m,1H),3.78(m ,1H),3.57(m,1H),3.51(m,1H),2.35(m,2H),2.09-1.91(m,2H),1.52(m,2H),1.33-1.25(m,28H),0.88(m,3H). 13 CNMR (100MHz, DMSO-d6) δ 172.5, 153.5, 152.7, 148.6, 141.2, 122.4, 99.1, 84.5, 74.6, 63.4, 37.3, 34.2, 32.3, 29.4, 28.7, 25.7, 22.3, 14.5. (Partial alkyl peak overlap) MSI-MS: 536.6 [M+H] + It was.

[0151] Example 22 N-(4-chloro-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)octadecamide [ka]

[0152] Using adenosine as the raw material, 15b was obtained by referring to the method of Example 15. Using 15b as the raw material, 6.28 g (10 mmol) of 15b was dissolved in ethyl acetate (40 mL) under a nitrogen gas atmosphere, and 10 mL (22 mmol) of a 37% solution of hydrochloric acid in triethylamine was added, stirred, and heated to 70° C. for about 8 hours. After TLC showed the completion of the reaction, it was cooled to room temperature, washed with saturated sodium bicarbonate solution until neutral, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was recrystallized with anhydrous methanol (30 mL) to obtain solid 22a (weight 1.95 g, yield 38%, MSI-MS: 515.2 [M+H] + ) was obtained.

[0153] Using 22a as a raw material, compound 22 was prepared according to the method of preparing 10a from 10b in Example 10, in combination with the deprotection method of 1a in Example 1, with a total yield of 79%. The detection results of the obtained compound 22 were HNMR (400MHz, DMSO-d6) δ 10.60 (s, 1H), 8.67 (s, 1H), 8.35 (s, 1H), 6.15 (d, 1H), 5.33 (d, 1H), 4.98 (m, 1H), 4.31 (m, 1H), 4.02 (m, 1H), 3.78 (m, 1H), 3.58 (m, 1H), 3.53 (m, 1H), 2.32 (m, 2H), 1.51 (m, 2H), 1.34-1.21 (m, 28H), 0.87 (m, 3H). 13 CNMR (100MHz, DMSO-d6) δ 171.4, 152.5, 151.6, 148.4, 140.5, 121.4, 98.7, 77.5, 74.3, 68.1, 59.4, 38.2, 33.7, 31.5, 28.1, 27.5, 25.4, 22.1, 14.6. (Partial alkyl peak overlap) MSI-MS: 553.2 [M+H] + It was.

[0154] Example 23 (((((4-hydroxy-5-(6-(isonicotinamido)-9H-purin-9-yl)tetrahydrofuran-2-yl)methoxy)methyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionic acid) [ka]

[0155] Compound 23a was prepared according to the protection method of 1b in Example 1 using compound 12 as the raw material, and compound 23 was prepared according to the process of adjusting 17 from 17a in Example 17 using 23a as the raw material. 17a in the reaction was replaced with 23a in an equivalent amount, and the total yield of the reaction was 81%. The detection result of the obtained compound 23 was as follows: 1 HNMR(400MHz,DMSO-d6)δ11.05,(s,1H),8,85(d,2H),8.34(s,1H),8.18(s,1H),8.00(d,2H),6.84(s,4H),6.14( d,1H),5.35(d,1H),4.01(m,1H),3.94(m,1H),3.86(s,2H),3.60-3.38(m,2H),2.06-1.84(m,2H),1.27(s,18H). 13 CNMR(100MHz,DMSO-d6)δ175.8,165.1,152.4,151.5,149.8,149.6,140.7,140.2,12 1.2,120.1,99.5,92.8,78.1,74.7,74.5,71.4,38.7,34.5,27.2.MSI-MS:679.6[M+H] + It was.

[0156] Example 24 Isopropyl((((4-hydroxy-5-(6-(isonicotinamido)-9H-purin-9-yl)tetrahydrofuran-2-yl)methoxy)methyl)(phenoxy)phosphoryl)alanine ester [ka]

[0157] Compound 23a was obtained according to the method of Example 23, and compound 24 was obtained from 23a according to the method of Example 7. Compound 1b was replaced with 23a in an equivalent amount, and the total yield was 85%. The detection result of the obtained compound 24 was as follows: 1HNMR(400MHz,DMSO-d6)δ11.08(s,1H),8.81(d,2H),8.36(s,1H),8.18(s,1H),8.02(d,2H),7.42(m,2H),7.21(m,3H),6.16 (d,1H),5.35(s,1H),4.92(m,1H),4.01-3.95(m,2H),3.65-3.54(m,5H),3.35(m,1H),2.04-1.84(m,2H),1.25-1.18(d,9H). 13 CNMR(100MHz,DMSO-d6)δ171.1,165.2,152.7,151.8,150.4,149.8,149.5,140.5,140.1,130.4,123.2 ,121.8,121.4,120.2,99.3,76.7,74.8,74.5,72.5,69.8,50.4,34.7,21.5,19.3.MSI-MS:640.6[M+H] + It was.

[0158] (Example 25) (5-(6-amino-9H-purin-9-yl)-3-azido-4-hydroxytetrahydrofuran-2-yl) dihydrogen methyl phosphate [ka]

[0159] Under argon gas atmosphere, 2.67 g (10.0 mmol) of adenosine and 35.78 g (22 mmol) of PPh were added to a round-bottom flask. Then, 18 mL of DMF was added, and the resulting mixture was stirred at room temperature until the solid was completely dissolved. The solution was cooled to 10°C, and 4.4 mL (2.2 mmol) of diisopropyl azodicarboxylate (DIAD) was added dropwise rapidly. The reaction mixture was allowed to reach room temperature. After stirring at 40°C for 2 h, the DMF was evaporated under reduced pressure to obtain a viscous liquid, and the mixture was partitioned between 150 mL of water and 100 mL of diethyl ether. The aqueous layer was collected and washed with 100 mL of diethyl ether. The organic phase was concentrated under reduced pressure, and the crude product was purified by column chromatography and eluted with DCM and acetone solvent mixture (3:1 to 2:3). A white solid compound 25a (weight 1.84 g, yield 74%) was obtained. The detection results of the obtained compound were as follows: 1 HNMR(400MHz,DMSO-d6)δ8.57(s,1H),8.36(s,1H),7.10(s,2H),6.37(d,1H),4.59(m,1H),3.95(s,1H),3.57-3.50(m,2H),2.62-2.55(m,2H). 13 CNMR(100MHz,DMSO-d6)δ156.3,152.5,149.9,140.5,119.5,90.4,82.6,63.5,60.8,59.6.MSI-MS:250.2[M+H] + It was.

[0160] 2.49 g (10 mmol) of compound 25a and 2.6 g (40 mmol) of sodium azide were added to a flask, and 20 mL of DMF was added as a solvent. The mixture was reacted at 120° C. for 16 hours. The progress of the reaction was monitored on a TLC plate. After the reaction was completed, water was added to quench the reaction, and the aqueous phase was extracted three times with ethyl acetate (180 mL). The organic phases were combined and washed with water. The solvent was removed under reduced pressure to obtain foamy solid 25b (weight 2.22 g, yield 76%, MSI-MS: 293.2 [M+H] + ) was obtained.

[0161] Starting from 25b, 25c was obtained according to the protection method for preparing 1b in Example 1. Starting from 25c, 1b was replaced with 25c in an equivalent amount according to the method in Example 2 to obtain compound 25 (yield 76%). The detection result of the obtained compound 25 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.57(s,1H),8.38(s,1H),7.09(s,2H),6.17(d,1H),5.3 5(d,1H),4.27-4.24(m,2H),4.19(s,2H),4.02(m,1H),3.81(m,1H),1.75(m,1H). 13 CNMR(100MHz,DMSO-d6)δ158.2,152.4,149.8,140.5,119.4,101.2,78.1,73.2,68.4,57.5.MSI-MS:373.2[M+H] + It was.

[0162] (Example 26) (((((5-(6-amino-9H-purin-9-yl)-3-azido-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionic acid) [ka]

[0163] Compound 25c was obtained according to the preparation method of Example 25, and compound 26a was obtained using 25c as a raw material according to the method of preparing compound 17 from compound 17a in Example 17, and compound 26 was obtained by the deprotection method of 1b in Example 1. 25c was replaced with 17a in an equivalent amount, and the total yield was 54%. The detection result of the obtained compound 26 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.34(s,1H),7.04(s,2H),6.82(s,4H),6.15(d,1H) ),5.35(d,1H),4.00(m,2H),3.85(s,2H),3.62-3.41(m,2H),1.84(m,1H),1.26(s,18H). 13CNMR(100MHz,DMSO-d6)δ175.4,157.3,152.5,149.7,140.2,119.5,99.8,92.8,79.0,72.3,71.5,71.1,56.5,38.8,27.6.MSI-MS:615.6[M+H] + It was.

[0164] Example 27 Isopropyl((((5-(6-amino-9H-purin-9-yl)-3-azido-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)(phenoxy)phosphoryl)alanine ester [ka]

[0165] Compound 25c was obtained according to the preparation method of Example 25, and 25c was used as the raw material to replace 1b with 25c in an equivalent amount according to the preparation method of Example 7 to obtain compound 27 (total yield 43%). The detection result of the obtained compound 27 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.34(s,1H),7.42(m,2H),7.21(m,3H),7.07(s,2H),6.15( d,1H),5.34(s,1H),4.95(d,1H),3.99(m,2H),3.76-3.34(m,6H),1.88(m,1H),1.27-1.16(d,9H). 13 CNMR(100MHz,DMSO-d6)δ171.4,156.4,152.1,150.1,149.5,140.3,130.5,121.2,120.5 ,119.4,100.5,78.4,72.8,72.0,71.5,69.1,57.2,50.5,21.5,19.1.MSI-MS:576.5[M+H] + It was.

[0166] (Example 28) Isopropyl((((3-azido-4-hydroxy-5-(6-(isonicotinamido)-9H-purin-9-yl)tetrahydrofuran-2-yl)methoxy)methyl)(phenoxy)phosphoryl)alanine ester [ka]

[0167] Using the obtained compound 27 as a raw material, compound 28a was obtained by referring to the protection method of 1a or 1b in Example 1, and using compound 28a as a raw material, cordycepin or 1b was substituted for compound 28a in an equivalent amount by referring to the preparation method of Example 12 to obtain compound 28 (total yield 37%). The detection result of the obtained compound 28 was as follows: 1 HNMR(400MHz,DMSO-d6)δ11.07(s,1H),8.83(d,2H),8.35(s,1H),8.20(s,1H),8.01(d,2H),7.41(m,2H),7.21( m,3H),6.15(d,1H),5.35(s,1H),4.94(d,1H),4.00(m,2H),3.72-3.32(m,6H),1.82(m,1H),1.24-1.17(d,9H). 13 CNMR(100MHz,DMSO-d6)δ171.5,164.5,152.1,151.6,150.1,149.8,149.4,140.7,140.3,130.2,122.2 ,121.5,121.1,120.4,100.4,78.7,72.5,72.1,71.5,69.2,57.4,50.8,21.6,19.2.MSI-MS:681.6[M+H] + It was.

[0168] (Example 29) 2-(6-amino-9H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3-yl valine salt [ka]

[0169] Compound 1c was obtained according to the protection method of preparing 1c in Example 1, and 1c was used as raw material for further reaction. 9.95 g (27.2 mmol) of compound 1c and 4.88 g (27.2 mmol) of CbzCl were added to 38.5 mL of toluene and 38.5 mL of water, and 4.70 g (34 mmol) of K2CO3 were added, and the mixture was vigorously stirred at a temperature below 25 ° C. After stirring at room temperature for 3 hours, 0.275 g (2.72 mmol) of triethylamine and 5.78 g of sodium chloride were added in sequence, and the mixture was stirred for another 30 minutes. The organic layer was separated and concentrated to obtain the desired oily product 29a (weight 13.1 g, yield 90%, MSI-MS: 522.6 [M + Na] + ) was obtained.

[0170] 4.99g (10mmol) of compound 29a and 1.36g (10mmol) of valinyl chloride were taken, and 60mL of anhydrous pyridine was added under ice bath environment, and the temperature was gradually raised to 40℃ and reacted for 10 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, water and ethyl acetate were added for extraction, and the organic phase was back-extracted. The organic phase was recovered and rotary evaporated to obtain an oily liquid 10b. The purified product 29b (5.03g, yield 84%, MSI-MS: 625.5[M+Na] + ) was obtained.

[0171] 5.99g of compound 29b (10mmol) was dissolved in 200mL of methanol. Then, 1.5g of ammonium formate (30mmol) and 0.75g of 10% Pd-C were added, and the reaction mixture was stirred at room temperature for 10 minutes, and then heated to reflux for 45 minutes. The mixture was filtered through diatomaceous earth, and the filtrate was evaporated to dryness to obtain compound 29c (4.41g, 95% yield). Using 29c as the raw material, compound 29 (90% yield) was obtained by deprotection according to the method of 1c in Example 1. The detection result of the obtained compound 29 was: 1HNMR(400MHz,DMSO-d6)δ8.89(s,2H),8.58(s,1H),8.37(s,1H),7.05(s,2H),6.73(d,1H),5.01(m,1H),4.9 2(s,1H),4.27(m,1H),3.75(m,1H),3.56-3.47(m,2H),2.38(m,1H),2.13(m,1H),1.89(m,1H),0.98(d,6H). 13 CNMR(100MHz,DMSO-d6)δ172.1,156.0,152.3,149.5,140.1,119.4,102.2,82.5,74.4,63.7,59.5,33.1,30.5,19.1.MSI-MS:351.3[M+H] + It was.

[0172] (Example 30) 2-(6-amino-9H-purin-9-yl)-5-(phosphonooxy)methyl)tetrahydrofuran-3-yl valine salt [ka]

[0173] Compound 29 was used as a raw material and reacted according to the method of Example 2 to obtain compound 30. 1b was substituted for 29 in an equivalent amount to obtain compound 30 (yield 84%). The detection result of the obtained compound 30 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.90(s,2H),8.56(s,1H),8.33(s,1H),7.04(s,2H),6.75(d,1H),5.0 0(m,1H),4.27-4.02(m,5H),3.73(m,1H),2.36(m,1H),2.16(m,1H),1.88(m,1H),0.96(d,6H). 13 CNMR(100MHz,DMSO-d6)δ172.4,156.3,152.5,149.8,140.2,119.5,101.2,76.1,74.2,68.7,59.3,32.1,30.4,19.0.MSI-MS:431.3[M+H] + It was.

[0174] (Example 31) 2-(6-(isonicotinamido)-9H-purin-9-yl)-5-(phosphono)methyl)tetrahydrofuran-3-yl valine salt [ka]

[0175] Using compound 30 as a raw material, 1a was replaced with compound 30 in an equivalent amount according to the preparation method of Example 12 to obtain compound 31 (yield 91%). The detection result of the obtained compound 31 was as follows: 1 HNMR(400MHz,DMSO-d6)δ11.08(s,1H),8.92(s,2H),8.82(d,2H),8.34(s,1H),8.20(s,1H),8.02(d,2H),6. 73(d,1H),5.03(m,1H),4.29-4.01(m,5H),3.75(m,1H),2.39(m,1H),2.15(m,1H),1.88(m,1H),0.97(d,6H). 13 CNMR(100MHz,DMSO-d6)δ171.4,164.5,152.3,151.1,149.9,149.7,140.5,140.1,1 23.7,121.5,101.7,77.1,74.5,68.2,59.8,32.4,30.8,19.2.MSI-MS:558.4[M+Na] + It was.

[0176] Example 32 (((5-(6-(isonicotinamido)-9H-purin-9-yl)-4-(pentyloxy)tetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid [ka]

[0177] Using compound 29 as a raw material, compound 1b was replaced with compound 29 in an equivalent amount according to the preparation method of Example 3 to obtain compound 32a (yield 77%). Using compound 32a as a substrate, 1a was replaced with compound 32a in an equivalent amount according to the preparation method of Example 12 to obtain compound 32 (yield 90%). The detection result of the obtained compound 32 was as follows:1 HNMR(400MHz,DMSO-d6)δ11.05(s,1H),8.91(s,2H),8.84(d,2H),8.35(s,1H),8.18(s,1H),8.01(d,2H),6.75(d,1H),5.01(m,1 H),4.81(s,2H),4.24(m,1H),3.95(m,1H),3.73(d,2H),3.60-3.35(m,2H),2.37(m,1H),2.14(m,1H),1.89(m,1H),0.96(d,6H). 13 CNMR(100MHz,DMSO-d6)δ171.5,165.5,152.4,151.1,149.8,140.7,140.1,123.5, 121.7,102.7,76.8,76.6,75.5,74.3,59.5,32.5,30.5,18.9.MSI-MS:550.4[M+H] + It was.

[0178] (Example 33) Methyl (((2-(6-amino-9H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3-yl)oxy)(phenoxy)phosphoryl)alaninate [ka]

[0179] Using compound 1c as a raw material, compound 1b was replaced with compound 1c in an equivalent amount according to the preparation method of Example 6 to obtain compound 33a (yield 62%), which was then reacted according to the deprotection method of 1c in Example 1 to obtain compound 33 (yield 90%). The detection result of the obtained compound 33 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.33(s,1H),7.42(m,2H),7.21(m,3H),7.08(s,2H), 6.15(d,1H),5.00(m,1H),4.02(m,1H),3.71-3.48(m,8H),2.24-1.98(m,2H),1.25(d,3H). 13CNMR(100MHz,DMSO-d6)δ172.1,156.1,152.4,149.9,149.6,140.3,131.0,121.5, 120.2,119.6,101.8,82.1,74.3,63.4,52.1,46.2,32.5,19.2.MSI-MS:439.4[M+H] + It was.

[0180] (Example 34) Methyl (((5-(hydroxymethyl)-2-(6-(isonicotinamido)-9H-purin-9-yl)tetrahydrofuran-3-yl)oxy(phenoxy)phosphoryl)alaninate [ka]

[0181] Compound 33a was obtained according to the process of Example 33. Compound 33a was used as the raw material, and 1a was replaced with compound 33a in an equivalent amount according to the preparation method of Example 12 to obtain compound 34 (total yield 83%). The detection result of the obtained compound 34 was as follows: 1 HNMR(400MHz,DMSO-d6)δ11.02(s,1 H),8.81(d,2H),8.34(s,1H),8.17(s,1H),8.02(d,2H),7.41(m,2H),7.23(m,3H),6.1 6(d,1H),4.98(m,1H),4.01(m,1H),3.78-3.47(m,8H),2.25-1.97(m,2H),1.27(d,3H). 13 CNMR(100MHz,DMSO-d6)δ171.4,163.8,152.1,151.5,150.4,149.8,149.7,140.8,140.5,130.2, 122.5,121.7,121.2,119.9,102.4,82.5,74.5,63.5,51.7,46.3,32.7,19.4.MSI-MS:598.5[M+H] + It was.

[0182] Example 35 (5-(2-((E)-2-bromocarbonyl)-6-(isonicotinamido)-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl) dihydrogen methyl phosphate [ka]

[0183] 2.51 g (10 mmol) of cordycepin was dissolved in dilute nitric acid, heated to 110 °C, and 1.26 g (5 mmol) of elemental iodine was added and reacted for 4 hours, and the reaction was monitored by TLC. After the reaction was completed, it was extracted with petroleum ether, the aqueous phase was collected, the organic phase solution was extracted with deionized water, and the aqueous phase was combined and rotary evaporated to obtain compound 35a (weight 2.72 g, yield 72%, MSI-MS: 378.2 [M + H] + ) was obtained.

[0184] 100 mL of anhydrous 1,4-dioxane was placed in a reactor, argon gas was introduced to protect the reactor, and the reactor was heated to 70°C. 0.11 g (0.5 mmol) of palladium acetate, 0.26 g (1 mmol) of triphenylphosphine, and 1.8 mL of triethylamine were added in sequence and stirred for about 30 minutes. 3.77 g (10 mmol) of compound 35a and 2.59 g (30 mmol) of methyl acrylate were added and reacted for 2 hours in an environment of 80°C. The reaction was then terminated, filtered, the filtrate was collected, allowed to stand, and further filtered to obtain compound 35b (weight 2.41 g, yield 72.7%, MSI-MS: 336.4 [M+H]) as a white cotton-like solid. + ) was obtained.

[0185] Compound 35b (1.00 g) was taken, 12 mL of sodium hydroxide solution (2 mol / L) was added dropwise, and the mixture was stirred at room temperature for 3 hours. Concentrated hydrochloric acid was then added dropwise in an ice bath until the pH reached 1, resulting in a large amount of white precipitate. The white solid 35c (weight 0.88 g, yield 92%, MSI-MS: 322.6 [M+H]) was obtained by suction filtration. + ) was obtained.

[0186] 3.21g (10mmol) of compound 35c was taken and added to 120mL of water, and heated to 100℃ with stirring. Then, 2.07g (15mmol) of anhydrous potassium carbonate was added, and 2.67g (15mmol) of NBS was dissolved in 22.5mL of acetone and 22.5mL of water, respectively, and the mixture was added dropwise to the flask, and the addition was completed within 135 minutes. After stirring for 3 hours, the reaction was stopped, half of the solvent was removed, and the mixture was placed in a refrigerator overnight. A large amount of needle-shaped brown crystals were precipitated and filtered by suction to obtain compound 35d (weight 1.78g, yield 50%, MSI-MS: 357.3 [M+H] + ) was obtained.

[0187] Compound 35d (1.78 g, 5 mmol) was taken, and 40.00 mL of anhydrous pyridine and 8.5 mL of acetic anhydride were added under ice bath conditions. The reaction was monitored by HPLC liquid phase, and the reaction was terminated after about 5 hours. The solvent was removed to obtain viscous liquid 35e (weight 1.76 g, yield 80%). Compound 35f (yield 95%) was prepared from compound 35e using the method of Example 16, and compound 35 (yield 72%) was obtained using the phosphorylation method of Example 2 and the deprotection method of Example 1. The detection result of the obtained compound 35 was as follows: 1 HNMR(400MHz,DMSO-d6)δ11.05(s,1H),8.84(d,2H),8.31(s,1H),8.02(d,2H),7.11(d,1H) ,6.70(d,1H),6.11(d,1H),5.35(s,1H),4.31-4.02(m,5H),3.76(m,1H),2.07-1.84(m,2H). 13 CNMR(100MHz,DMSO-d6)δ164.5,152.6,151.8,149.8,149.7,140.9,140.3,1 34.5,124.3,123.4,121.5,98.5,74.5,74.3,68.5,34.7.MSI-MS:542.2[M+H] + It was.

[0188] (Example 36) 3-((5-(6-amino-2-fluoro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-5-(hydroxymethyl)cyclopentane-1,2-diol [ka]

[0189] Compound 17a was obtained according to the method of Example 17, and 17a was used as the raw material to prepare according to the preparation method of Example 8, and 2-chloro-3-hydroxypropylvaline was replaced with 3-chloro-5-(hydroxymethyl)cyclopentane-1,2-diol in an equivalent amount to obtain compound 36a, which was then deprotected to obtain compound 36 (total yield 41%). The detection result of the obtained compound 36 was: 1 HNMR(400MHz,DMSO-d6)δ8.37(s,1H),7.01(s,2H),6.15(d,1H),5.92(s,1H),5.37(d,1H),4.38(s,1 H),4.25(s,1H),4.02(m,2H),3.84(m,1H),3.65-3.31(m,6H),2.07-1.82(m,2H),1.70-1.44(m,3H). 13 CNMR(100MHz,DMSO-d6)δ156.1,153.5,149.8,140.3,119.5,99.2,83.5,81.2,76.2,75.7,74.8,73.1,64.1,34.9,34.7,31.5.MSI-MS:416.8[M+H] + It was.

[0190] (Example 37) Preparation of the compound: (((((5-(6-amino-9H-purin-9-yl)-2-azido-4-hydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphoryl)bis(oxy)bis(methylene)bis(2,2-dimethylpropionic acid) [ka]

[0191] 3.65g (10mmol) of compound 1b and 3.44g (20mmol) of metachloroperbenzoic acid were added to a reaction flask, and 100mL of anhydrous dichloromethane was added to dissolve the compound. 2.13g (15mmol) of diphosphorus pentoxide was added separately under an ice bath environment, and the temperature was gradually raised to 40°C and reacted for 3 hours. After the reaction was completed, the compound was filtered, and 100mL of saturated sodium bicarbonate was added to perform a quenching reaction. The compound was further washed twice with 50mL of dichloromethane and back-extracted twice with 60mL of saturated sodium bicarbonate. The organic phases were combined, concentrated under reduced pressure, and subjected to column chromatography to obtain compound 37a (weight 1.49g, yield 41%). The detection results of the obtained compound were as follows: 1 HNMR(400MHz,DMSO-d6)δ8.56(s,1H),8.36(s,1H),7.10(s,2H),6.16(d,1 H),4.02(s,1H),2.75-2.45(m,2H),2.13(d,2H),0.99(s,9H),0.22(s,6H). 13 CNMR(100MHz,DMSO-d6)δ156.5,152.5,149.8,140.4,119.6,102.5,85.9,69.3,52.7,42.5,30.8,25.8,0.MSI-MS:364.5[M+H] + It was.

[0192] 3.63g (10mmol) of compound 37a and 2.6g (40mmol) of sodium azide were added to a flask, and 20mL of DMF was added as a solvent. The mixture was reacted for 16 hours under an environment of 120°C. The progress of the reaction was monitored on a TLC plate. After the reaction was completed, water was added to quench the reaction, and the aqueous phase was extracted three times with ethyl acetate (180mL). The organic phases were combined, washed with water, and the solvent was removed under reduced pressure to obtain foamy solid 37b (weight 2.77g, yield 68%). Compound 37 was obtained according to the method of preparing compound 17 from compound 17a in Example 17 using 37b as a raw material. 37b was replaced with 17a in an equivalent amount, and the total yield was 50%. The detection result of the obtained compound 37 was as follows: 1HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.34(s,1H),7.07(s,2H),6.82(d,4H),6.15(d,1H), 5.34(d,1H),4.02(m,1H),3.84(d,2H),3.61-3.42(m,2H),2.11-1.80(m,2H),1.28(d,18H). 13 CNMR(100MHz,DMSO-d6)δ175.2,156.3,152.6,149.9,140.2,119.4,100.3,93.1,90.2,81.0,71.5,68.8,38.6,36.2,27.4.MSI-MS:637.6[M+Na] + It was.

[0193] (Example 38) Preparation of the compound (5-(6-amino-2-((E)-2-bromocarbonyl-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methyl dihydrogen phosphate, reaction steps are as follows: [ka]

[0194] Compound 38 (total yield 24%) was obtained from compound 2 by using the method of Example 35 using compound 2 as a raw material. The detection result of the obtained compound 38 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.35(s,1H),7.09(d,1H),6.95(s,2H),6.68(d,1H), 6.15(d,1H),5.35(s,1H),4.27-4.02(m,5H),3.74(m,1H),2.07-1.84(m,2H). 13 CNMR(100MHz,DMSO-d6)δ156.1,152.3,149.8,140.2,134.9,124.2,119.5,98.7,74.9,74.3,68.1,34.5.MSI-MS:458.3[M+N] + It was.

[0195] (Example 39) Preparation of compound: (5-(6-amino-8-(isopropylamino)-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methyl dihydrogen phosphate. The preparation steps are as follows: [ka]

[0196] Compound 39 was prepared using compound 1b as a raw material. First, 3.65 g (10 mmol) of compound 1b and 1.18 g (20 mmol) of 2-propylamine were added to a reaction flask, 60 mL of dioxane was added as a solvent, and the mixture was refluxed for 20 hours under an environment of 80 ° C., and the progress of the reaction was monitored by TLC. After the reaction was completed, the reaction solution was concentrated, and then extracted twice with 50 mL of water and 50 mL of ethyl acetate. The organic phases were combined and concentrated under reduced pressure to obtain an oily liquid, and the pale yellow oily liquid obtained by purifying it by column chromatography was compound 39a (weight 1.44 g, yield 34%).

[0197] Compound 39 was prepared according to the preparation method in Example 2 using compound 39a as the raw material, and 1b was replaced with 39a in an equivalent amount, with a yield of 91%. The detection result of the obtained compound 39 was: 1 HNMR(400MHz,DMSO-d6)δ8.15(s,1H),7.05(s,2H),6.15(d,1H),5.98(s,1H), 5.38(s,1H),4.28-3.95(m,6H),3.75(m,1H),2.05-1.81(m,2H),1.18(d,6H). 13 CNMR(100MHz,DMSO-d6)δ153.1,152.3,151.5,149.8,118.5,99.7,74.8,74.5,68.5,46.5,34.3,23.5.MSI-MS:389.3[M+H] + It was.

[0198] (Example 40) Preparation of compound: 3-((5-(6-amino-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-5-(hydroxymethyl)cyclopentane-1,2-diol, the preparation steps are as follows: [ka]

[0199] Compound 1b was used as the raw material, and was prepared according to the preparation method of Example 8, and 2-chloro-3-hydroxypropylvaline was replaced with 3-chloro-5-(hydroxymethyl)cyclopentane-1,2-diol in an equivalent amount to obtain compound 40 (total yield 46%). The detection result of the obtained compound 40 was as follows: 1 HNMR(400MHz,DMSO-d6)δ8.58(s,1H),8.37(s,1H),7.04(s,2H),6.15(d,1H),5.91(s,1H),5.37(d,1H),4. 37(s,1H),4.25(s,1H),4.00(m,2H),3.82(m,1H),3.63-3.32(m,6H),2.08-1.81(m,2H),1.71-1.45(m,3H). 13 CNMR(100MHz,DMSO-d6)δ156.3,153.4,149.7,140.2,119.4,99.5,83.4,81.1,76.1,75.5,74.9,73.6,64.4,34.9,34.7,31.6.MSI-MS:382.3[M+H] + It was.

[0200] (Example 41) Antitumor effects of modified cordycepin derivatives on experimental hepatocellular carcinoma Preparation of mouse liver cancer transplant model and drug evaluation. 2.5×10 5 The hepatoma cells Hep-1-6 were suspended in 100 μL of PBS and inoculated into the outer side of the left thigh of C57BL / 6j nude mice. The transplanted tumor grew to about 100 mm after one week. 3When the size of the tumor reached 10, the mice were randomly divided into 12 groups of 10 mice each, which were the blank group, the cordycepin control group, and the drug group (compounds 2, 3, 5, 7, 15, 24, 27, 31, 35, 40). The blank group was forcibly administered nutrients such as corn syrup and DMSO as a solvent control every other day, while the cordycepin control group and the drug group were forcibly administered compound drugs (500ug / time / mouse) every other day, and observed for 14 consecutive days. The changes in the size of the liver cancer tumors in the two groups of mice were observed, and the mice were sacrificed after the experiment was completed, and blood and corresponding tissue specimens were collected. The tumor bodies were immediately photographed and weighed, and a portion of the tumor tissue was collected and fixed in formalin solution for further examination. The two perpendicular diameters (length and width) of the transplanted tumor were measured every 2 days using a caliper, and the size of the transplanted tumor was calculated. The results of the tumor volume change experiment and the results of the blank group, the cordycepin control group, and the tumors treated with the drug group within 14 days are shown in Figure 1. Formula: Tumor volume (mm 3 )=1 / 2×(length×width) 2 The experimental results showed that when the drugs were used alone, they could significantly inhibit the growth of mouse tumors, and all of them were more effective than the cordycepin control group. Compound 24 had a better anti-liver cancer tumor effect (900 mm 3 From 100mm 3 The difference in the tumor-cell survival time was reduced to less than 0.05 (P<0.001), indicating that the compound drug group had the effect of killing liver cancer tumor cells or activating tumor immunity.

[0201] (Example 42) Experimental antitumor effects of modified cordycepin derivatives on small cell lung cancer The antitumor dose-response curves of cordycepin and compounds 1, 2, 4, 5, 7, 8, 10, 12, 16, 18, 21, 23, 24, 35, and 38 were analyzed in small cell lung cancer cell lines H1048, H446, and H69 using the MTT assay. 50) were calculated, and the results are summarized in the table below. The results of the in vitro antitumor experiments showed that compared with cordycepin, the effective concentrations of the modified cordycepin derivatives against tumor cells were decreased, among which compounds 35 and 38 had strong in vitro killing effects against the three types of cells, but also had high toxicity, followed by compounds 18 and 21. The IC of cordycepin in small cell lung cancer cell lines H1048 and H446 50 In both cases, the effective concentrations were higher than 100 μM, and it was possible that a large amount of drug would be required to achieve the effect in practical use.

[0202] [Table 1]

[0203] (Example 43) Determination of the maximum tolerated dose of modified cordycepin derivatives in an in vivo experiment with zebrafish 240 3dpf wild-type AB strain zebrafish were randomly selected with 30 fish per well and collected into 8-well plates with a volume of 3mL of fish culture water per well, cordycepin was diluted according to the maximum lethal dose, dissolved in DMSO, and added to each well in a concentration gradient (the maximum lethal dose was diluted 100-fold, and 8 concentration gradient values ​​were tested in the middle), and a normal control group, a solvent control group G (5% glucose) and a solvent control group D (DMSO) were set up. During the experiment, each group of zebrafish was cultured in an environment of 35°C, and after treating the zebrafish with the test product until 5dpf, the lethality and developmental malformation of the zebrafish at different doses were examined, and the maximum tolerated dose (MTD) of the test product to normal zebrafish was determined at a concentration with a survival rate of more than 90% and a teratogenicity rate of less than 20%. Cordycepin was selected as the control group, and further, Compound 2, Compound 5, Compound 6, Compound 7, Compound 9, Compound 11, Compound 13, Compound 20, Compound 24, Compound 25, Compound 28, Compound 35, Compound 36, Compound 37, and Compound 39 were selected to test the MTD in zebrafish according to the above method, and the test results are shown in Table 2.

[0204] [Table 2]

[0205] (Example 44) Inhibitory effects of modified cordycepin derivatives on zebrafish small cell lung cancer in vivo We established zebrafish tumor-bearing models of three types of small cell lung cancer, H69, H446, and H1048 cells, each of which had a tumor size of 1×10 6 H69, H446, H1048 cell suspensions at a concentration of 1000 mL were prepared in serum-free cell medium, and 5 mL of red fluorescent dye (CM-DiI) cell labeling solution was added per 1 mL of cell suspension, mixed gently, incubated at 37 °C for 20 min, centrifuged at 1500 rpm for 5 min, the supernatant was removed, and serum-free medium was added again to resuspend the cells. The above steps were repeated twice to obtain the desired labeled cell suspensions, which were then transplanted into zebrafish yolk sacs by microinjection, with approximately 100 cells transplanted per animal, to create a zebrafish-human small cell lung cancer susceptibility line transplant tumor model. The zebrafish injected with ovarian cancer cells were cultured at 35 °C until 3 dpf.

[0206] To test the effect of cordycepin and prepared compounds 2, 5, 6, 7, 9, 11, 13, 20, 24, 25, 28, 35, 36, 37, 39 on the growth of anti-small cell lung cancer in zebrafish in vivo, and inhibit tumor zebrafish model up to 3 dpf, zebrafish with good consistency of transplanted tumor were selected under a microscope and randomly assigned to 6-well plates with 30 per well. At 3 dpf, the above compounds were diluted at MTD dose, dissolved in DMSO and added to each well, and normal control group, model control group and solvent control group (DMSO) were set up with a volume of 3 mL of fish water per well. After the zebrafish in each experimental group were further cultured at 35℃ until 5dpf, 10 zebrafish per experimental group were randomly selected to be observed under a fluorescent microscope, photographed and saved, and analyzed using Nikon NIS-Elementsl3.10 high-end image processing software to calculate the fluorescence intensity (S) of the zebrafish transplanted tumors. The inhibition results are shown in Table 3 below, among which the inhibition results of the control group, cordycepin group, compound 24 and compound 35 groups in the transplanted zebrafish model of H446 small cell lung cancer are shown in Figure 2. The growth inhibition effect of the candida injection solution and paclitaxel alone on the zebrafish-human ovarian cancer susceptible line was calculated by the total fluorescence intensity, and the formula is as follows: Tumor growth inhibition effect (%) = S (model control group) - S (drug group) / S (model control group) × 100%

[0207] [Table 3]

[0208] (Example 45) Experimental inhibition of colon cancer by modified cordycepin derivatives Preparation of mouse colon cancer transplant model and drug evaluation. 2.5×10 6 MC-38 colon cancer cells were suspended in 100 μL of PBS and inoculated into the outer side of the left thigh of C57BL / 6j nude mice. One week later, the transplanted tumor grew to approximately 100 mm 3When the mice reached the size of 100 mm, they were randomly assigned to 7 groups, with 8 mice per group, as follows: control group IgG, compound 16 drug + IgG group, PD-1 antibody treatment group, CTLA4 monoclonal antibody treatment group, compound 16 drug + PD-1 combination treatment group, and compound 16 drug + CTLA4 monoclonal antibody combination treatment group. Control group IgG: immunoglobulin G (IgG) (500 μg / dose / animal) was forcefully administered intragastrically every day for 28 consecutive days. Cordycepin + IgG: immunoglobulin G (IgG) and cordycepin (both 500 μg / dose / animal) were forcefully administered intragastrically every day for 28 consecutive days. Compound 16 drug + IgG: immunoglobulin G (IgG) and compound 16 drug (both 500 μg / dose / animal) were forcefully administered intragastrically every day for 28 consecutive days. Antibody treatment group: antibody (500 μg / dose / animal) was administered intragastrically every 4 days for 28 consecutive days. Compound 16 drug + antibody combination treatment group: Immunoglobulin (500 μg / dose / mouse) was forcibly administered intragastrically every day for 28 consecutive days, and Compound 16 drug (500 μg / dose / mouse) was administered intragastrically every 4 days for 28 consecutive days. After the experiment was completed, the mice were sacrificed, blood and corresponding tissue specimens were collected, the tumor body was immediately imaged and weighed, and a portion of the tumor tissue was taken and fixed in formalin solution for further examination. The two perpendicular diameters (length and width) of the implanted tumor were measured every 2 days using a caliper, and the size of the implanted tumor was calculated using the formula: tumor volume (mm 3 )=1 / 2×(length×width) 2 The volume was calculated by the above method, and the results of observing the change in size of the colon cancer tumor in the mice of the 7 groups are shown in Figure 3. After 28 days, the experimental results showed that cordycepin had an anti-proliferative effect on colon cancer (the size was 1300 mm 3 From 730mm 3 Compound 16, when used alone, had a significant antitumor growth effect compared to cordycepin (size reduced to 1300 mm 3 From 480mm 3 (P<0.01), and the immune checkpoint inhibitors PD-1 and CTLA4 used alone had a significant anti-tumor growth effect (size reduced to 1300 mm 3 From 448mm 3When compound 16 was used in combination with immune checkpoint inhibitors PD-1 and CTLA4, the synergistic effect of the two significantly enhanced the antitumor effect (the size was reduced to 1300 mm 3 From 260mm 3 The results showed that the number of

[0209] (Example 46) Experimental inhibitory effects of modified cordycepin derivatives on melanoma. Preparation of mouse melanoma transplant model and drug evaluation: 2.5×10 5 B16-F10 melanoma cells were suspended in 100 μL of PBS and inoculated into the outer side of the left thigh of C57BL / 6j nude mice. One week later, the transplanted tumor grew to approximately 100 mm 3 When the size of the tumor reached 100 mm, the mice were randomly assigned to 10 groups, with 10 mice per group, as follows: blank control group, cordycepin group, compound 13 group, compound 14 group, compound 17 group, compound 19 group, compound 22 group, compound 26 group, compound 29 group, and compound 34 group. The blank control group was forcibly administered nutrients such as corn syrup and DMSO every day for 24 consecutive days. The compound groups were forcibly administered the corresponding compounds (500ug / time / mouse) every other day for 24 consecutive days, and the size changes of the melanomas of the 10 groups of mice were observed every 4 days in the early period and every 2 days in the late period. After the experiment was completed, the mice were sacrificed, and blood and the corresponding tissue specimens were collected. The tumor body was immediately photographed and weighed, and a portion of the tumor tissue was taken and fixed in formalin solution for further examination. The two perpendicular diameters (length and width) of the transplanted tumor were measured every 2 days using a caliper, and the size of the transplanted tumor was calculated, using the formula: tumor volume (mm 3 )=1 / 2×(length×width) 2 The volume was calculated by the above method, and the experimental results are shown in Figure 4. The experimental results showed that cordycepin itself inhibited the growth of melanoma (1760 mm 3 From 600mm 3 ), and modified cordycepin derivatives were more effective than cordycepin (1760 mm 3 From 560mm 3Compounds 19 and 34, when used alone, significantly inhibited the growth of mouse tumors (1760 mm 3 From about 240 mm 3 The results showed that the compound had the effect of killing tumor cells.

[0210] (Example 46-1) Compound 27 is used to inhibit melanoma in the above manner with the same level of inhibitory effect as compound 26.

[0211] (Example 47) Experimental inhibitory effects of modified cordycepin derivatives on ovarian cancer. Preparation of mouse ovarian cancer transplant model and drug evaluation. 2.5×10 6 ID8 ovarian cancer cells were suspended in 100 μL of PBS and inoculated into the outer side of the left thigh of C57BL / 6j nude mice. After about one week, the transplanted tumor grew to about 100 mm 3When the mice reached the size of 10 mm, they were randomly assigned to 5 groups, with 10 mice per group, as the control group, the cordycepin group, the compound 24 drug group, the PD-1 + TIM3 antibody treatment group, and the compound 24 drug + PD-1 + TIM3 antibody treatment group. Control group: Immunoglobulin G (IgG) (500 μg / dose / mouse) and PBS were forcefully administered intragastrically every day for 15 consecutive days. Cordycepin group: Cordycepin (500 μg / dose / mouse) was mixed with PBS and forcefully administered intragastrically every day for 15 consecutive days. Compound 24 drug group: Compound 24 (500 μg / dose / mouse) was mixed with PBS and forcefully administered intragastrically every day for 15 consecutive days. PD-1 + TIM3 antibody treatment group: Antibody PD-1 + TIM3 (each 500 μg / dose / mouse) was administered intragastrically every 4 days for 15 consecutive days. Compound 24 drug + PD-1 + TIM3 antibody treatment group: Immunoglobulin and compound 24 drug (500 μg / time / animal) were forcibly administered intragastrically every day for 15 consecutive days, and antibody PD-1 + TIM3 (each 500 μg / time / animal) was administered intragastrically every 4 days for 15 consecutive days. The size changes of ovarian tumors in the mice of the 5 groups were observed every 2 days, and after the end of the experiment, the mice were sacrificed, blood and corresponding tissue specimens were collected, the tumor body was immediately photographed and weighed, and a part of the tumor tissue was taken and fixed in formalin solution for further examination. The two perpendicular diameters (length and width) of the implanted tumor were measured every 2 days using a caliper, and the size of the implanted tumor was calculated, using the formula: tumor volume (mm 3 )=1 / 2×(length×width) 2 The volume was calculated by the above method, and the experimental results are shown in Figure 5. The experimental results show that cordycepin has an ovarian cancer anti-proliferation effect (size reduced by 1.5 times in 15 days), compound 24 used alone has a more significant anti-tumor proliferation effect than cordycepin (size reduced by about 2 times in 15 days), immune checkpoint inhibitors PD-1 and CTLA4 used alone have a significant anti-tumor proliferation effect (size reduced by more than 2 times in 15 days), and compound 24 and immune checkpoint inhibitors PD-1 and CTLA4 used together significantly enhance the anti-tumor effect (size reduced by about 4 times in 15 days).

[0212] (Example 48) Experimental inhibitory effects of modified derivatives of cordycepin on gastric cancer cells in vitro The antitumor dose-response curves of cordycepin and the 40 obtained compounds were analyzed in gastric cancer cell lines AGS and BGC-823 using MTT assay. 50 ) was calculated, and the results are summarized in Table 4 below. The results of the in vitro antitumor experiments showed that the effective concentration of the modified cordycepin derivatives against tumor cells was reduced compared to cordycepin.

[0213] [Table 4]

[0214] (Example 49) Experimental inhibitory effects of modified cordycepin derivatives on in vivo tumor growth in a mouse model of gastric cancer Preparation of mouse gastric cancer transplant model and drug evaluation: 2×10 7 BGC-823 gastric cancer cells were suspended in 100 μL of PBS, and the anterior chest wall of a C57BL / 6j nude mouse was disinfected with 75% ethanol. The area where the apical pulsation was most obvious was touched with the hand, and the cells were inoculated into the left ventricle about 3 mm from the left edge of the sternum in the second intercostal space. After about one week, the transplanted tumor grew to about 100 mm 3When the tumor size reached 100 mg / kg, the mice were randomly divided into 5 groups, with 10 mice per group, and were classified as the control group, cordycepin group, compound 16, 18, and 24 drug groups, respectively. Control group: Nutrients such as corn syrup and DMSO were administered intragastrically every day as a solvent control group. Cordycepin group: Cordycepin (500 μg / time / mouse) was mixed with DMSO and administered intragastrically every day. Compound drug group: Compound (500 μg / time / mouse) was mixed with DMSO and administered intragastrically every day for 18 consecutive days. The size changes of the gastric tumors in the 5 groups of mice were observed every 2 days, and after the experiment was completed, the mice were sacrificed, blood and corresponding tissue specimens were collected, and the tumor bodies were immediately photographed and weighed. The weighing experimental results are shown in Figure 6. The experimental results showed that cordycepin could effectively inhibit the growth of gastric tumors, with a reduction of about 4 times compared to the control group (reduced from 1.73 g to 0.46 g); modified cordycepin had a more pronounced effect than cordycepin, with the compound 24 group showing a reduction of about 12 times compared to the control group (reduced from 1.73 g to 0.14 g).

[0215] (Example 50) Experimental inhibitory effects of modified cordycepin derivatives on in vivo tumor growth in a mouse model of pancreatic cancer Preparation of mouse pancreatic cancer transplant model and drug evaluation: Pancreatic cancer Pan02-luc cells were placed in DMEM medium containing 10% fetal bovine serum and placed in a CO2 incubator at 37 °C. Pancreatin EDTA was digested and passaged every 2-3 days. When the cells reached the desired number, the cells in the logarithmic growth phase were harvested and diluted to 1 × 10 in medium. 7The cells were resuspended in 1000μg / mL. C57BL / 6j nude mice were bred under pathogen-free conditions, and after the mice had grown for 6 weeks, 200μL of pancreatic cancer Pan02-uc cells were subcutaneously injected into the right dorsal axilla of each mouse. The appearance of transplanted tumors within about 1 week indicated that the model was successfully created. The mice were randomly assigned to 5 groups, 10 mice per group, as the control group, cordycepin group, compound 11, 16, and 24 drug groups, respectively. Control group: Nutrients such as corn syrup and DMSO were administered intragastrically every day as a solvent control group. Cordycepin group: Cordycepin (500μg / time / mouse) was mixed with DMSO and administered intragastrically every day. Compound drug group: Compound (500μg / time / mouse) was mixed with DMSO and administered intragastrically every day. The administration was continued for 24 consecutive days. Thereafter, the survival rates of the five groups of mice were observed daily, and the median survival times were examined, as shown in Figure 7. The experimental results showed that administration of cordycepin alone significantly extended the survival time of tumor-bearing mice (increased from 41 to 52 days), the compound group further extended the survival time of tumor-bearing mice based on cordycepin (increased from 41 to more than 58 days), and compound 24 was the most effective, extending the survival time of tumor-bearing mice by almost half (increased from 41 to 76 days).

[0216] It should be noted that the above is merely a preferred embodiment of the present invention, and those skilled in the art can make various improvements and modifications without departing from the spirit of the present invention, and these improvements and modifications should also be considered as within the protection scope of the present invention. Each component not specified in this embodiment can be realized by using conventional technology.

[0217] (Additional Note) (Appendix 1) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is R1 10 -R1 11 , or R1 14-R1 16 R2 is selected from hydrogen or an azide group, R3 is selected from hydrogen, R4 is selected from a cyano group, R5 is selected from hydrogen, a bromovinyl group, a mercapto group, a methyl group, fluorine, or chlorine, R6 is selected from an amino group, and R7 is selected from hydrogen or an isopropylamino group.

[0218] (Appendix 2) A cordycepin derivative represented by formula I as described in Appendix 1, characterized in that the cordycepin derivative represented by formula I is compound 16, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. [ka]

[0219] (Appendix 3) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is R1 10 -R1 11 , or R1 14 -R1 16 R2 is selected from hydrogen or an azide group, R3 is selected from hydrogen, R4 is selected from a hydroxyl group, R5 is selected from hydrogen, a bromovinyl group, a mercapto group, a methyl group, a fluorine group, or a chlorine group, and R6 is selected from R6 15 and R7 is selected from hydrogen or an isopropylamino group.

[0220] (Appendix 4) A cordycepin derivative represented by formula I as described in Appendix 3, characterized in that the cordycepin derivative represented by formula I is compound 24 or compound 23, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka]

[0221] (Appendix 5) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is selected from hydrogen, R2 is selected from hydrogen or an azide group, R3 is selected from hydrogen, and R4 is R4 6 or R4 7 R5 is selected from hydrogen, a bromovinyl group, a mercapto group, a methyl group, fluorine, or chlorine; R6 is selected from R6 15 and R7 is selected from hydrogen or an isopropylamino group.

[0222] (Appendix 6) A cordycepin derivative represented by formula I as described in Appendix 5, characterized in that the cordycepin derivative represented by formula I is compound 34, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. [ka]

[0223] (Appendix 7) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is R1 10 -R1 11 , or R1 14 -R1 16R2 is selected from hydrogen or an azide group, R3 is selected from hydrogen, R4 is selected from a hydroxyl group, R5 is selected from a mercapto group, R6 is selected from an amino group, and R7 is selected from hydrogen or an isopropylamino group.

[0224] (Appendix 8) The cordycepin derivative represented by formula I according to Appendix 7, characterized in that the cordycepin derivative represented by formula I is compound 19, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. [ka]

[0225] (Appendix 9) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is R1 10 -R1 11 , or R1 14 -R1 16 R2 is selected from hydrogen or an azide group, R3 is selected from hydrogen, R4 is selected from a hydroxyl group, R5 is selected from fluorine, R6 is selected from an amino group, and R7 is selected from hydrogen or an isopropylamino group.

[0226] (Appendix 10) A cordycepin derivative represented by formula I according to Appendix 9, characterized in that the cordycepin derivative represented by formula I is compound 20 or compound 17, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. [ka]

[0227] (Appendix 11) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is R1 10 -R1 11 , or R1 14 -R1 16 R2 is selected from hydrogen or an azide group, R3 is selected from fluorine, R4 is selected from a hydroxyl group, R5 is selected from hydrogen, a bromovinyl group, a mercapto group, a methyl group, fluorine, or chlorine, R6 is selected from an amino group, and R7 is selected from hydrogen or an isopropylamino group.

[0228] (Appendix 12) A cordycepin derivative represented by formula I according to Appendix 11, characterized in that the cordycepin derivative represented by formula I is compound 18, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. [ka]

[0229] (Appendix 13) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is R1 10 -R1 11 , or R1 14 -R1 16R2 is selected from hydrogen or an azide group, R3 is selected from an azide group, R4 is selected from a hydroxyl group, R5 is selected from hydrogen, a bromovinyl group, a mercapto group, a methyl group, fluorine, or chlorine, R6 is selected from an amino group, and R7 is selected from hydrogen or an isopropylamino group.

[0230] (Appendix 14) A cordycepin derivative represented by formula I as described in Appendix 13, characterized in that the cordycepin derivative represented by formula I is compound 26 or compound 27, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka]

[0231] (Appendix 15) A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. [ka] (Wherein, R1 is selected from hydrogen, R2 is selected from hydrogen or an azide group, R3 is selected from hydrogen, and R4 is R4 5 R5 is selected from hydrogen, a bromovinyl group, a mercapto group, a methyl group, fluorine, or chlorine, R6 is selected from an amino group, and R7 is selected from hydrogen or an isopropylamino group.

[0232] (Appendix 16) The cordycepin derivative represented by formula I according to Appendix 15, characterized in that the cordycepin derivative represented by formula I is compound 29, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. [ka]

[0233] (Appendix 17) A pharmaceutical composition comprising the cordycepin derivative according to any one of Appendices 1 to 16, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof, and at least one immune checkpoint inhibitor, wherein the immune checkpoint inhibitor is preferably selected from PD-1 and / or CTLA4 monoclonal antibodies, and preferably, the mass ratio of the cordycepin derivative, or the pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof to the immune checkpoint inhibitor is 1:0.2-10, preferably 1:0.2-8, preferably 1:0.2-5, preferably 1:0.2-3, preferably 1:0.2-2, preferably 1:0.5-1.5, preferably 1:0.8-1.2, and preferably 1:1.

[0234] (Appendix 18) 18. The pharmaceutical composition according to claim 17, wherein the dosage form of the pharmaceutical composition is selected from tablets, pellets, capsules, pills, syrups, disintegrants, injections, sustained release agents, or kits.

[0235] (Appendix 19) Use of a cordycepin derivative according to any one of appendices 1 to 16, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof, or a pharmaceutical composition according to appendices 17 or 18, in the preparation of a product for preventing and treating a disease associated with mutation of cellular dysfunction in a mammal or human, wherein the disease associated with mutation of cellular dysfunction is preferably a tumor, and the tumor preferably includes gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, colorectal cancer, esophageal cancer, prostate cancer, melanoma, glioma, and ovarian cancer, and the tumor is preferably any one of gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, colorectal cancer, melanoma, and ovarian cancer.

[0236] (Appendix 20) A method for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R1 is R1 10 -R1 11 , or R1 14 -R1 16 In the case where the compound IR-1 is selected from the group represented by the formula (I), the method for preparing the cordycepin derivative represented by the formula (I) comprises chemically reacting the compound IR-1 as a raw material in an organic solvent to obtain the cordycepin derivative represented by the formula (I); [ka] (In formula IR-1, R2 to R7 are the same as R2 to R7 in formula I, or each is independently selected from a protecting group.) In formula I, R1 is R1 14 -R1 15 In the case where the cordycepin derivative is selected from the group represented by the formula I, preferably, the method for preparing the cordycepin derivative represented by the formula I is to react the compound IR-1 with nitrophenyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide and / or tetrahydrofuran using tert-butylmagnesium chloride as a catalyst, the substitution being substituted with one or more functional groups of an aryloxy group and an amide amino acid group, preferably, the ratio of the amounts of the compound IR-1, the modifying agent, the catalyst and the organic solvent used is 1 mmol: 1-3 mmol: 1-2 mmol: 9-15 mL, preferably, the reaction temperature is 20-40° C., and preferably, the reaction time is 2-5 hours; R1 in formula I is R1 10 -R1 11 , or R1 16In the case where the cordycepin derivative is selected from the groups represented by the formula I, preferably, the method for preparing the cordycepin derivative represented by the formula I is characterized in that the compound IR-1 is reacted with p-toluenesulfonyloxymethyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide using NaH as a catalyst, the substitution being substituted with one or more functional groups of either an aryloxy group or an amide amino acid group, preferably, the ratio of the amounts of the compound IR-1, the modifying agent, the catalyst and the organic solvent used is 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:100-200 mL, preferably 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:150 mL, preferably, the reaction temperature is -20-0°C, and preferably, the reaction time is 0.5-6 hours. Preparation method.

[0237] (Appendix 21) A method for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In the case where R3 in formula I is selected from fluorine or azide group, the method for preparing the cordycepin derivative represented by formula I includes chemically reacting compound IR-3 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I; [ka] (In formula IR-3, R1, R2, R5 to R7 are the same as R1, R2, R5 to R7 in formula I, or each is independently selected from a protecting group.) In formula I, when R3 is selected from fluorine, the preparation method of the cordycepin derivative represented by formula I is preferably carried out by reacting compound IR-3 with trifluoromethanesulfonic anhydride in pyridine and dichloromethane to obtain intermediate IR-3a, 5-(6-amino-9H-purin-9-yl)-4-hydroxy-2-(hydroxymethyl)tetrahydrofuran-3-yl trifluoromethanesulfonic acid ester or its derivative, and then by substitution reaction of intermediate IR-3a with hydrofluoric acid and sulfur trifluoride in ethyl acetate to obtain the cordycepin derivative represented by formula I. In the preparation method of intermediate IR-3a, the ratio of the amount of compound IR-3, trifluoromethanesulfonic anhydride, pyridine and dichloromethane used is preferably 1. mmol:1-1.5 mmol:0.15-0.2 mL:10-20 mL, preferably the reaction temperature is -5-5°C, preferably 0°C, and preferably the reaction time is 1-3 hours; in the preparation method of the cordycepin derivative represented by formula I, preferably intermediate IR-3a is subjected to a substitution reaction with 37% hydrofluoric acid or diethylaminosulfur trifluoride or a triethylamine solution of hydrochloric acid, preferably the ratio of the amount of intermediate IR-3a, 37% hydrofluoric acid or diethylaminosulfur trifluoride or a triethylamine solution of hydrochloric acid to ethyl acetate is 1 mmol:2-3 mmol:4-10 mL, preferably the reaction temperature is 60-80°C, and preferably the reaction time is 8-10 hours; In the case where R3 in formula I is selected from an azide group, the preparation method of the cordycepin derivative represented by formula I is preferably carried out by subjecting compound IR-3 to a cyclization reaction in N,N-dimethylformamide under the catalysis of triphenylphosphine and diisopropyl azodicarboxylate to obtain intermediate IR-3b, 4-(6-amino-9H-purin-9-yl)-3,6-dioxane[3.1.0]hexan-2-yl)methanol or a derivative thereof, and then subjecting the obtained intermediate IR-3b to a ring-opening reaction with sodium azide in dimethylformamide to obtain the cordycepin derivative represented by formula I. In the preparation method of the intermediate IR-3b, preferably, the compound IR-3, triphenylphosphine, diisopropyl azodicarboxylate are catalyzed. The ratio of the amounts of sodium azide and N,N-dimethylformamide used is 10.0 mmol: 16-28 mmol: 1.6-2.8 mmol: 15-50 mL, preferably 10.0 mmol: 22 mmol: 2.2 mmol: 15-50 mL, preferably the cyclization reaction temperature is 10-60°C, preferably the cyclization reaction time is 1-5 hours, preferably the method for preparing the cordycepin derivative represented by formula I is characterized in that the ratio of the amounts of intermediate IR-3b, sodium azide and dimethylformamide used is 1 mmol: 4-5 mmol: 2-5 mL, preferably the ring-opening reaction temperature is 100-120°C, preferably the ring-opening reaction time is 12-16 hours; Preparation method.

[0238] (Appendix 22) A method for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R4 is a cyano group, R4 5 ~R4 7 In the case where the compound IR-4 is selected from the group represented by the formula (I), the preparation method of the cordycepin derivative represented by the formula (I) comprises chemically reacting the compound IR-4 as a raw material in an organic solvent to obtain the cordycepin derivative represented by the formula (I); [ka] (In formula IR-4, R1 to R3 and R5 to R7 are the same as R1 to R3 and R5 to R7 in formula I, or each is independently selected from a protecting group.) In the case where R4 in formula I is selected from a cyano group, the preparation method of the cordycepin derivative represented by formula I is preferably carried out by stirring compound IR-4, trifluoromethanesulfonic acid, and trimethylsilyl triflate in dichloromethane under an environment of −50 to −30° C., and then reacting with trimethylsilyl cyanide and triethylamine, and preferably, the ratio of the amounts of compound IR-4, trifluoromethanesulfonic acid, trimethylsilyl triflate, and dichloromethane used is 10 mmol:0.8 to 1.8 mL: Preferably, the stirring time is 20 to 40 minutes, preferably 30 minutes. Preferably, the ratio of the amounts of compound IR-4, trimethylsilyl cyanide, and triethylamine used is 10 mmol: 3.4 to 4.3 g: 3 to 4 mL. Preferably, the reaction temperature is 20 to 30° C., preferably room temperature. Preferably, the reaction time is 2 to 4 hours. In formula I, R4 is R4 5 In the case where the compound IR-4 is selected from the group represented by the formula (I), preferably, the method for preparing the cordycepin derivative represented by the formula (I) comprises reacting the compound IR-4 with an amino acid chloride in anhydrous pyridine to obtain the cordycepin derivative represented by the formula (I), preferably, the ratio of the amounts of the compound IR-4, the amino acid chloride and the pyridine used is 10 mmol: 5-15 mmol: 50-100 mL, preferably 10 mmol: 10 mmol: 50-100 mL, preferably, the reaction temperature is 20-60°C, preferably 40°C, preferably, the reaction time is 6-20 hours, In formula I, R4 is R4 7In the case where the cordycepin derivative is selected from the group represented by the formula I, preferably, the method for preparing the cordycepin derivative represented by the formula I is to react the compound IR-4 with the modifying agent p-toluenesulfonyloxymethylphenyl phosphate substituted with an amino acid alkyl ester in anhydrous N,N-dimethylformamide using NaH as a catalyst, preferably, the ratio of the amounts of the compound IR-4, the modifying agent, NaH and anhydrous N,N-dimethylformamide used is 0.1 mol: 0.1-0.15 mol: 0.2-0.03 mol: 100-200 mL, preferably 0.1 mol: 0.1-0.15 mol: 0.2-0.03 mol: 150 mL, preferably, the reaction temperature is -20-0°C, and preferably, the reaction time is 0.5-6 hours, In formula I, R4 is R4 6 In the case where the compound IR-4 is selected from the group represented by the formula I, the preparation method of the cordycepin derivative represented by the formula I is preferably characterized in that the compound IR-4 is reacted with nitrophenyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide and / or tetrahydrofuran using tert-butylmagnesium chloride as a catalyst, the substitution being substituted with one or more functional groups of an aryloxy group and an alkyl acid methyloxy group, the ratio of the amounts of the compound IR-4, the modifying agent, the catalyst and the organic solvent used is preferably 1 mmol: 1-3 mmol: 1-2 mmol: 9-15 mL, the reaction temperature is preferably 20-40° C., and the reaction time is preferably 2-5 hours. Preparation method.

[0239] (Appendix 23) A method for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In the formula I, when R5 is selected from a mercapto group or fluorine, the method for preparing the cordycepin derivative represented by formula I includes chemically reacting compound IR-5 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I; [ka] (In formula IR-5, R1 to R4 and R6 to R7 are the same as R1 to R4 and R6 to R7 in formula I, or each is independently selected from a protecting group.) In the formula I, when R5 is selected from a mercapto group, the method for preparing the cordycepin derivative represented by the formula I is preferably carried out by reacting the compound IR-5 with hydrogen peroxide in acetic acid to obtain intermediate IR-5d, 1N-oxide-3'-deoxyadenosine or its derivative, and heating the intermediate IR-5d in an aqueous hydrochloric acid solution to reflux to obtain intermediate IR-5e, 5-amino-N'-hydroxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1H-isopropyl ether or its derivative. midazole-4-carboxamide or its derivatives are obtained; intermediate IR-5e is dissolved in water and reacted with Raney nickel catalyst under hydrogen atmosphere to obtain intermediate IR-5f, 5-amino-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1H-imidazole-4-carboxamide or its derivatives; intermediate IR-5f is reacted with methanol, pyridine and carbon disulfide to obtain cordycepin derivatives represented by formula I; preferably, The method for preparing intermediate IR-5d comprises reacting compound IR-5 with acetic acid in aqueous hydrogen peroxide, preferably the concentration of the aqueous hydrogen peroxide is 20% to 40%, and preferably 30%, preferably the ratio of the amounts of compound IR-5, aqueous hydrogen peroxide, and acetic acid used is 1 mmol: 2-3 mol: 1-6 mL, preferably the reaction temperature is 30-50° C., preferably the reaction time is 2-4 days, and preferably the method for preparing intermediate IR-5e comprises reacting compound IR-5 with acetic acid in aqueous hydrogen peroxide, and preferably the concentration of the aqueous hydrogen peroxide is 20% to 40%, and preferably 30%, preferably the ratio of the amounts of compound IR-5, aqueous hydrogen peroxide, and acetic acid used is 1 mmol: 2-3 mol: 1-6 mL, preferably the reaction temperature is 30-50° C., preferably the reaction time is 2-4 days, Intermediate IR-5d is heated to reflux in an aqueous solution, preferably the concentration of the aqueous hydrochloric acid solution is 1-5 mol / L, preferably 3 mol / L, preferably the ratio of the amount of intermediate IR-5d to the aqueous hydrochloric acid solution used is 1 mmol:3-5 mL, preferably the heating to reflux time is 10-30 minutes, and in the method for preparing intermediate IR-5f, preferably the ratio of the amount of intermediate IR-5e to Raney nickel and water used is 1 mmol:0.08-0.In the method for preparing the cordycepin derivative represented by formula I, preferably, the ratio of the amount of the intermediate IR-5f to the amount of methanol, pyridine and carbon disulfide used is 1 mmol:5-10 mL, preferably, the ratio of the amount of the methanol, pyridine and carbon disulfide used is 4:3-7:0.5-3.5, preferably 4:5:2, preferably, the reaction temperature is 30-50°C, preferably 40°C, and preferably, the reaction time is 3-5 days. [ka] In formula I, when R5 is selected from fluorine, the preparation method of the cordycepin derivative represented by formula I is preferably carried out by nitro-derivating compound IR-5 to obtain nitrated intermediate IR-5g, 2-(6-amino-2-nitro-1,6-dihydro-9H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3-ol, which is then subjected to a substitution reaction to obtain the cordycepin derivative represented by formula I; the preparation method of the intermediate IR-5g is preferably carried out by reacting compound IR-5 with tetrabutylammonium nitrate in dichloromethane under the catalysis of trifluoroacetic anhydride; Preferably, the method for preparing the cordycepin derivative represented by formula I comprises reacting 5g of nitrated intermediate IR-5g with tetrabutylammonium fluoride in acetonitrile, preferably, the ratio of the amounts of nitrated intermediate IR-5g, tetrabutylammonium fluoride, and acetonitrile is 1 mmol: 1.3-1.5 mmol: 30-50 mL, the reaction temperature is -5-5°C, preferably 0°C, and the reaction time is preferably 20-30 minutes. Preparation method. [ka]

[0240] (Appendix 24) A method for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R6 is R6 15 In the case where the cordycepin derivative represented by formula I is selected from the group represented by the formula: [ka] (In formula IR-6, R1 to R5 and R7 are the same as R1 to R5 and R7 in formula I, or each is independently selected from a protecting group.) The method for preparing cordycepin derivatives represented by formula I is preferably characterized in that in anhydrous pyridine, compound IR-6 is reacted with a substituted acid chloride, the substitution being substituted with a pyridyl group, the ratio of compound IR-6, the substituted acid chloride and anhydrous pyridine used is preferably 1 mmol: 1-2 mmol: 5-10 mL, the reaction temperature is preferably 0-60°C, and the reaction time is preferably 2-20 hours. Preparation method.

Claims

1. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 1】 (In the formula, R １ is R １ １０ -R １ １１ , or R １ １４ -R １ １６ R ２ is selected from hydrogen or an azide group; R ３ is selected from hydrogen, R ４ is selected from a cyano group; R ５ is selected from hydrogen, bromovinyl, mercapto, methyl, fluorine, or chlorine; R ６ is selected from an amino group; R ７ is selected from hydrogen or an isopropylamino group.

2. The cordycepin derivative of formula I according to claim 1, characterized in that the cordycepin derivative of formula I is compound 16, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 2】

3. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 3】 (In the formula, R １ is R １ １０ -R １ １１ , or R １ １４ -R １ １６ R ２ is selected from hydrogen or an azide group; R ３ is selected from hydrogen, R ４ is selected from a hydroxyl group, R ５ is selected from hydrogen, bromovinyl, mercapto, methyl, fluorine, or chlorine; R ６ is R ６ １５ R ７ is selected from hydrogen or an isopropylamino group.

4. The cordycepin derivative of formula I according to claim 3, characterized in that the cordycepin derivative of formula I is compound 24 or compound 23, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 4】

5. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 5】 (In the formula, R １ is selected from hydrogen, R ２ is selected from hydrogen or an azide group; R ３ is selected from hydrogen, R ４ is R ４ ６ Or R ４ ７ R ５ is selected from hydrogen, bromovinyl, mercapto, methyl, fluorine, or chlorine; R ６ is R ６ １５ R ７ is selected from hydrogen or an isopropylamino group.

6. The cordycepin derivative of formula I according to claim 5, characterized in that the cordycepin derivative of formula I is compound 34, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 6】

7. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 7】 (In the formula, R １ is R １ １０ -R １ １１ , or R １ １４ -R １ １６ R ２ is selected from hydrogen or an azide group; R ３ is selected from hydrogen, R ４ is selected from a hydroxyl group, R ５ is selected from mercapto groups, R ６ is selected from an amino group; R ７ is selected from hydrogen or an isopropylamino group.

8. The cordycepin derivative of formula I according to claim 7, characterized in that the cordycepin derivative of formula I is compound 19, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 8】

9. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 9】 (In the formula, R １ is R １ １０ -R １ １１ , or R １ １４ -R １ １６ R ２ is selected from hydrogen or an azide group; R ３ is selected from hydrogen, R ４ is selected from a hydroxyl group, R ５ is selected from fluorine, R ６ is selected from an amino group; R ７ is selected from hydrogen or an isopropylamino group.

10. The cordycepin derivative of formula I according to claim 9, characterized in that the cordycepin derivative of formula I is compound 20 or compound 17, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 10】

11. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 11】 (In the formula, R １ is R １ １０ -R １ １１ , or R １ １４ -R １ １６ R ２ is selected from hydrogen or an azide group; R ３ is selected from fluorine, R ４ is selected from a hydroxyl group, R ５ is selected from hydrogen, bromovinyl, mercapto, methyl, fluorine, or chlorine; R ６ is selected from an amino group; R ７ is selected from hydrogen or an isopropylamino group.

12. The cordycepin derivative of formula I according to claim 11, characterized in that the cordycepin derivative of formula I is compound 18, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 12】

13. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 13】 (In the formula, R １ is R １ １０ -R １ １１ , or R １ １４ -R １ １６ R ２ is selected from hydrogen or an azide group; R ３ is selected from an azide group; R ４ is selected from a hydroxyl group, R ５ is selected from hydrogen, bromovinyl, mercapto, methyl, fluorine, or chlorine; R ６ is selected from an amino group; R ７ is selected from hydrogen or an isopropylamino group.

14. The cordycepin derivative of formula I according to claim 13, characterized in that the cordycepin derivative of formula I is compound 26 or compound 27, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 14】

15. A cordycepin derivative represented by formula I, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or metabolite thereof. 【Chemistry 15】 (In the formula, R １ is selected from hydrogen, R ２ is selected from hydrogen or an azide group; R ３ is selected from hydrogen, R ４ is R ４ ５ R ５ is selected from hydrogen, bromovinyl, mercapto, methyl, fluorine, or chlorine; R ６ is selected from an amino group; R ７ is selected from hydrogen or an isopropylamino group.

16. The cordycepin derivative of formula I according to claim 15, characterized in that the cordycepin derivative of formula I is compound 29, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof. 【Chemistry 16】

17. A pharmaceutical composition comprising the cordycepin derivative according to any one of claims 1 to 16, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof, and at least one immune checkpoint inhibitor, preferably the immune checkpoint inhibitor is selected from PD-1 and / or CTLA4 monoclonal antibodies, and preferably the mass ratio of the cordycepin derivative, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof to the immune checkpoint inhibitor is 1:0.2-10, preferably 1:0.2-8, preferably 1:0.2-5, preferably 1:0.2-3, preferably 1:0.2-2, preferably 1:0.5-1.5, preferably 1:0.8-1.2, and preferably 1:

1.

18. 18. The pharmaceutical composition according to claim 17, wherein the dosage form of the pharmaceutical composition is selected from a tablet, a pellet, a capsule, a pill, a syrup, a disintegrant, an injection, a sustained release agent, or a kit.

19. Use of the cordycepin derivative according to any one of claims 1 to 16, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug or metabolite thereof, or the pharmaceutical composition according to claim 17 or 18, in the preparation of a product for preventing and treating a disease associated with a mutation in a cellular dysfunction in a mammal or human, wherein the disease associated with a mutation in a cellular dysfunction is preferably a tumor, and the tumor preferably includes gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, colorectal cancer, esophageal cancer, prostate cancer, melanoma, glioma, and ovarian cancer, and the tumor is preferably any one of gastric cancer, pancreatic cancer, liver cancer, small cell lung cancer, colorectal cancer, melanoma, and ovarian cancer.

20. A process for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R １ R １ １０ -R １ １１ , or R １ １４ -R １ １６ In the case where the compound I-R-1 is selected from the group represented by the formula (I), the method for preparing the cordycepin derivative represented by the formula (I) comprises chemically reacting the compound I-R-1 as a raw material in an organic solvent to obtain the cordycepin derivative represented by the formula (I); 【Chemistry 17】 (In formula I-R-1, R ２ ~R ７ is R in formula I ２ ~R ７ or each independently is selected from a protecting group. In formula I, R １ R １ １４ -R １ １５ In the case where the cordycepin derivative is selected from the group represented by the formula I, preferably, the method for preparing the cordycepin derivative represented by the formula I is to react the compound I-R-1 with the nitrophenyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide and / or tetrahydrofuran using tert-butylmagnesium chloride as a catalyst, the substitution being substituted with one or more functional groups of an aryloxy group and an amide amino acid group, preferably, the ratio of the amounts of the compound I-R-1, the modifying agent, the catalyst and the organic solvent used is 1 mmol: 1-3 mmol: 1-2 mmol: 9-15 mL, preferably, the reaction temperature is 20-40° C., and preferably, the reaction time is 2-5 hours; R in Formula I １ R １ １０ -R １ １１ , or R １ １６ In the case where the cordycepin derivative is selected from the group represented by the formula I, preferably, the method for preparing the cordycepin derivative represented by the formula I is characterized in that the compound I-R-1 is reacted with p-toluenesulfonyloxymethyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide using NaH as a catalyst, the substitution being substituted with one or more functional groups of either an aryloxy group or an amide amino acid group, preferably, the ratio of the amounts of the compound I-R-1, the modifying agent, the catalyst and the organic solvent used is 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:100-200 mL, preferably, 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:150 mL, preferably, the reaction temperature is -20-0°C, and preferably, the reaction time is 0.5-6 hours. Preparation method.

21. A process for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R ３ is selected from fluorine or azide group, the method for preparing the cordycepin derivative represented by formula I includes chemically reacting compound I-R-3 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I, 【Chemistry 18】 (In formula IR-3, R １ , R ２ , R ５ ~R ７ is R in formula I １ , R ２ , R ５ ~R ７ or each independently is selected from a protecting group. In formula I, R ３ is selected from fluorine, the preparation method of the cordycepin derivative represented by formula I is preferably a method of reacting compound I-R-3 with trifluoromethanesulfonic anhydride in pyridine and dichloromethane to obtain intermediate I-R-3a, 5-(6-amino-9H-purin-9-yl)-4-hydroxy-2-(hydroxymethyl)tetrahydrofuran-3-yl trifluoromethanesulfonate or a derivative thereof, and a substitution reaction of intermediate I-R-3a with hydrofluoric acid and sulfur trifluoride in ethyl acetate to obtain the cordycepin derivative represented by formula I. In the preparation method of intermediate I-R-3a, the ratio of the amounts of compound I-R-3, trifluoromethanesulfonic anhydride, pyridine and dichloromethane used is preferably 1m mol: 1-1.5 mmol: 0.15-0.2 mL: 10-20 mL, preferably the reaction temperature is -5-5°C, preferably 0°C, and preferably the reaction time is 1-3 hours; in the preparation method of the cordycepin derivative represented by formula I, preferably the intermediate I-R-3a is subjected to a substitution reaction with 37% hydrofluoric acid or diethylaminosulfur trifluoride or a triethylamine solution of hydrochloric acid, and preferably the ratio of the amount of the intermediate I-R-3a, 37% hydrofluoric acid or diethylaminosulfur trifluoride or a triethylamine solution of hydrochloric acid, and ethyl acetate is 1 mmol: 2-3 mmol: 4-10 mL; preferably the reaction temperature is 60-80°C, and preferably the reaction time is 8-10 hours; In formula I, R ３ is selected from an azide group, the preparation method of the cordycepin derivative represented by formula I is preferably carried out by subjecting a compound I-R-3 to a cyclization reaction in N,N-dimethylformamide under the catalysis of triphenylphosphine and diisopropyl azodicarboxylate to obtain an intermediate I-R-3b, 4-(6-amino-9H-purin-9-yl)-3,6-dioxane[3.1.0]hexan-2-yl)methanol or a derivative thereof, and then subjecting the obtained intermediate I-R-3b to a ring-opening reaction with sodium azide in dimethylformamide to obtain a cordycepin derivative represented by formula I. In the preparation method of the intermediate I-R-3b, preferably, Preferably, the ratio of the amounts of the intermediate I-R-3b, sodium azide and N,N-dimethylformamide used is 10.0 mmol: 16-28 mmol: 1.6-2.8 mmol: 15-50 mL, and more preferably, the cyclization reaction temperature is 10-60° C., and the cyclization reaction time is 1-5 hours. Preferably, in the method for preparing the cordycepin derivative represented by formula I, the ratio of the amounts of the intermediate I-R-3b, sodium azide and dimethylformamide used is 1 mmol: 4-5 mmol: 2-5 mL, and preferably, the ring-opening reaction temperature is 100-120° C., and the ring-opening reaction time is 12-16 hours. Preparation method.

22. A process for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R ４ is a cyano group, R ４ ５ ~R ４ ７ In the case where the compound I-R-4 is selected from the group represented by the formula (I), the preparation method of the cordycepin derivative represented by the formula (I) comprises chemically reacting the compound I-R-4 as a raw material in an organic solvent to obtain the cordycepin derivative represented by the formula (I); 【Chemistry 19】 (In formula IR-4, R １ ~R ３ , R ５ ~R ７ is R in formula I １ ~R ３ , R ５ ~R ７ or each independently is selected from a protecting group. In formula I, R ４ is selected from a cyano group, the preparation method of the cordycepin derivative represented by formula I is preferably carried out by stirring compound I-R-4, trifluoromethanesulfonic acid, and trimethylsilyl triflate in dichloromethane under an environment of −50 to −30° C., and then reacting with trimethylsilyl cyanide and triethylamine, and preferably, the ratio of the amounts of compound I-R-4, trifluoromethanesulfonic acid, trimethylsilyl triflate, and dichloromethane used is 10 mmol:0.8 to 1.8 mL:2.2 3.2 mL:90-110 mL, preferably 10 mmol:1.3 mL:2.7 mL:100 mL, preferably the stirring time is 20-40 minutes, preferably 30 minutes, preferably the ratio of the amounts of compound I-R-4, trimethylsilyl cyanide, and triethylamine used is 10 mmol:3.4-4.3 g:3-4 mL, preferably the reaction temperature is 20-30° C., preferably room temperature, preferably the reaction time is 2-4 hours, In formula I, R ４ R ４ ５ In the case where the compound I-R-4 is selected from the group represented by the formula (I), the preparation method of the cordycepin derivative represented by the formula I is preferably carried out by reacting the compound I-R-4 with an amino acid chloride in anhydrous pyridine to obtain the cordycepin derivative represented by the formula I, preferably, the ratio of the amounts of the compound I-R-4, the amino acid chloride and the pyridine used is 10 mmol: 5-15 mmol: 50-100 mL, preferably 10 mmol: 10 mmol: 50-100 mL, preferably, the reaction temperature is 20-60°C, preferably 40°C, preferably, the reaction time is 6-20 hours, In formula I, R ４ R ４ ７ In the case where the compound I-R-4 is selected from the group represented by the formula (I), the preparation method of the cordycepin derivative represented by the formula (I) is preferably carried out by reacting the compound I-R-4 with the modifying agent p-toluenesulfonyloxymethylphenyl phosphate substituted with an amino acid alkyl ester in anhydrous N,N-dimethylformamide using NaH as a catalyst, and the ratio of the amounts of the compound I-R-4, the modifying agent, NaH and anhydrous N,N-dimethylformamide used is preferably 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:100-200 mL, and preferably 0.1 mol:0.1-0.15 mol:0.2-0.03 mol:150 mL, and the reaction temperature is preferably -20 to 0°C, and the reaction time is preferably 0.5 to 6 hours, In formula I, R ４ R ４ ６ In the case where the cordycepin derivative is selected from the group represented by the formula I, preferably, the method for preparing the cordycepin derivative represented by the formula I is characterized in that the compound I-R-4 is reacted with nitrophenyl phosphate substituted with a modifying agent in anhydrous N,N-dimethylformamide and / or tetrahydrofuran using tert-butylmagnesium chloride as a catalyst, the substitution being substituted with one or more functional groups of an aryloxy group and an alkyl acid methyloxy group, preferably, the ratio of the amounts of the compound I-R-4, the modifying agent, the catalyst and the organic solvent used is 1 mmol: 1-3 mmol: 1-2 mmol: 9-15 mL, preferably, the reaction temperature is 20-40° C., and preferably, the reaction time is 2-5 hours. Preparation method.

23. A process for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R ５ is selected from a mercapto group or fluorine, the method for preparing the cordycepin derivative represented by formula I includes chemically reacting compound I-R-5 as a raw material in an organic solvent to obtain the cordycepin derivative represented by formula I, 【Chemistry 20】 (In formula IR-5, R １ ~R ４ , R ６ ~R ７ is R in formula I １ ~R ４ , R ６ ~R ７ or each independently is selected from a protecting group. In Formula I, R ５ is selected from a mercapto group, the preparation method of the cordycepin derivative represented by formula I is preferably carried out by reacting compound I-R-5 with hydrogen peroxide in acetic acid to obtain intermediate I-R-5d, 1N-oxide-3'-deoxyadenosine or a derivative thereof, heating intermediate I-R-5d in an aqueous hydrochloric acid solution under reflux to obtain intermediate I-R-5e, 5-amino-N'-hydroxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1H-imidazole-4-carboxamide or a derivative thereof, dissolving intermediate I-R-5e in water, and dissolving it in Raney reacting with nickel catalyst under hydrogen atmosphere to obtain intermediate I-R-5f, 5-amino-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1H-imidazole-4-carboxamide or its derivative; reacting intermediate I-R-5f with methanol, pyridine and carbon disulfide to obtain cordycepin derivative represented by formula I; preferably, the method for preparing intermediate I-R-5d comprises reacting compound I-R-5 with acetic acid in hydrogen peroxide solution, preferably, the concentration of hydrogen peroxide solution is 20% to 40%, preferably 30%; Preferably, the ratio of the amounts of the compound I-R-5, hydrogen peroxide solution, and acetic acid used is 1 mmol: 2-3 mol: 1-6 mL, preferably, the reaction temperature is 30-50° C., preferably, the reaction time is 2-4 days, preferably, the method for preparing the intermediate I-R-5e comprises heating the intermediate I-R-5d in an aqueous hydrochloric acid solution under reflux, preferably, the concentration of the aqueous hydrochloric acid solution is 1-5 mol / L, preferably 3 mol / L, preferably, the ratio of the amounts of the intermediate I-R-5d and the aqueous hydrochloric acid solution used is 1 mmol: 3-5 mL, preferably, The heating reflux time is 10 to 30 minutes, and in the preparation method of the intermediate I-R-5f, preferably, the ratio of the amount of the intermediate I-R-5e, Raney nickel, and water used is 1 mmol: 0.08 to 0.2 g: 10 to 20 mL, preferably, the reaction temperature is 50 to 70°C, and preferably, the reaction time is 2 to 5 days. In the preparation method of the cordycepin derivative represented by formula I, preferably, the ratio of the amount of the intermediate I-R-5f, methanol, pyridine, and carbon disulfide used is 1 mmol: 5 to 10 mL, and preferably, the methanol,The ratio of the amounts of pyridine and carbon disulfide used is 4:3-7:0.5-3.5, preferably 4:5:2; the reaction temperature is preferably 30-50° C., preferably 40° C.; the reaction time is preferably 3-5 days; 【Chemistry 21】 In formula I, R ５ is selected from fluorine, the preparation method of the cordycepin derivative represented by formula I is preferably a method of preparing the cordycepin derivative represented by formula I by nitro-derivating compound I-R-5 to obtain nitrated intermediate I-R-5g, 2-(6-amino-2-nitro-1,6-dihydro-9H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3-ol, which is then subjected to a substitution reaction to obtain the cordycepin derivative represented by formula I; the preparation method of the intermediate I-R-5g is preferably a method of preparing the intermediate I-R-5 by reacting compound I-R-5 with tetrabutylammonium nitrate in dichloromethane with the catalyst of trifluoroacetic anhydride; Preferably, the method for preparing the cordycepin derivative represented by formula I comprises reacting nitrated intermediate I-R-5g with tetrabutylammonium fluoride in acetonitrile, preferably, the ratio of the amounts of nitrated intermediate I-R-5g, tetrabutylammonium fluoride, and acetonitrile is 1 mmol: 1.3-1.5 mmol: 30-50 mL, the reaction temperature is −5° C. to 5° C., preferably 0° C., and the reaction time is preferably 20-30 minutes. Preparation method. 【Chemical 22】

24. A process for preparing the cordycepin derivative according to any one of claims 1 to 16, comprising the steps of: In formula I, R ６ R ６ １５ In the case where the compound I-R-6 is selected from the group represented by the formula (I), the method for preparing the cordycepin derivative represented by the formula (I) comprises chemically reacting the compound I-R-6 as a raw material in an organic solvent to obtain the cordycepin derivative represented by the formula (I); 【Chemistry 23】 (In formula IR-6, R １ ~R ５ , R ７ is R in formula I １ ~R ５ , R ７ or each independently is selected from a protecting group. The method for preparing cordycepin derivatives represented by formula I is preferably characterized in that in anhydrous pyridine, compound I-R-6 is reacted with a substituted acid chloride, said substitution being substituted with a pyridyl group, the ratio of compound I-R-6, substituted acid chloride and anhydrous pyridine used is preferably 1 mmol: 1-2 mmol: 5-10 mL, the reaction temperature is preferably 0-60°C, and the reaction time is preferably 2-20 hours; Preparation method.

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