Cordycepin derivative as well as preparation method and application thereof

Abstract

The invention discloses a cordycepin derivative as well as a preparation method and application thereof. The structural general formula of the cordycepin derivative is # imgabs0. The cordycepin derivative provided by the invention is structurally modified by adopting a special group, so that not only is the stability of the cordycepin derivative improved, but also the biological activities of bacteriostasis, inflammation resistance, oxidation resistance, tumor resistance and the like are remarkably improved. Besides, the cordycepin derivative provided by the invention has certain selectivity on strains, especially on certain beneficial bacteria, in the aspect of bacteriostasis, and the application range of the cordycepin derivative is expanded.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a cordycepin derivative and a preparation method and use thereof. Background Art

[0002] Nucleoside drugs can block the growth of RNA or DNA chains and thus exert their efficacy. Cordycepin is the first nucleoside antibiotic isolated from fungi and has a variety of pharmacological activities, including antibacterial, anti-inflammatory, antioxidant, anti-tumor, immunomodulatory and prevention of hyperlipidemia. Cordycepin and its derivatives have received extensive attention and exploration in the field of biomedicine, but the types of cordycepin derivatives currently developed are relatively limited. In addition, due to their specific structure, cordycepin and its derivatives have poor chemical stability and cannot be widely used. The N-acyl cordycepin derivatives synthesized by Wei et al. can protect its amino group and improve its biological activity; Shimada et al. reported that replacing hydrogen at the 4' position with fluorine can delay its metabolism and reduce its cytotoxicity. The types of cordycepin derivatives currently obtained are limited, and although adding certain side groups to cordycepin can delay deamination, it still cannot completely block the deamination of adenosine deaminase (ADA enzyme), so it is still necessary to develop cordycepin derivatives with higher biological activity and chemical stability. Summary of the invention

[0003] In view of the above technical problems, the present invention provides a cordycepin derivative and a preparation method and use thereof. The cordycepin derivative provided by the present invention has both strong stability and high biological activity, providing more drug options for clinical application.

[0004] To achieve the above object, the technical solution adopted by the present invention is:

[0005] In one aspect, the present invention provides a cordycepin derivative, the general structural formula of which is shown in formula (I):

[0006]

[0007] In formula (I):

[0008] R1 and R2 are each independently selected from any one of alkoxycarbonyl and H;

[0009] R3 is selected from any one of an alkoxycarbonyl group, a silicon group, a substituted benzenesulfonyl group and H;

[0010] R4 is selected from any one of a thiol-substituted heterocyclic ring, a thiol-substituted aromatic ring and H;

[0011] R5 is any one of silicon group and H; R1 to R5 are not H at the same time.

[0012] In the technical solution of the present invention, the structural formula of the alkoxycarbonyl group is The Z1 group is selected from any one of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms, a substituted or unsubstituted cycloalkyl group having 3 to 10 carbon atoms, a substituted or unsubstituted cycloalkenyl group having 3 to 10 carbon atoms, a substituted or unsubstituted aryl group having 6 to 30 carbon atoms, a substituted or unsubstituted aryloxy group having 6 to 30 carbon atoms and a substituted or unsubstituted arylthio group having 6 to 30 carbon atoms, and is preferably any one of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms.

[0013] In the technical solution of the present invention, the structural formula of the silicon-based The Z2 group, the Z3 group, and the Z4 group are each independently selected from any one of hydrogen, -F, -Cl, -Br, hydroxyl, cyano, nitro, amidino, hydrazine, hydrazone, alkyl having 1 to 10 carbon atoms, alkenyl having 2 to 10 carbon atoms, alkynyl having 2 to 10 carbon atoms, alkoxy having 1 to 10 carbon atoms, cycloalkyl having 3 to 10 carbon atoms, cycloalkenyl having 3 to 10 carbon atoms, aryl having 6 to 30 carbon atoms, aryloxy having 6 to 30 carbon atoms, and arylthio having 6 to 30 carbon atoms, and are preferably any one of alkyl having 1 to 10 carbon atoms.

[0014] In the technical solution of the present invention, the structural formula of the substituted benzenesulfonyl group is C is any one of an aryl group having 6 to 30 carbon atoms and a heteroaryl group having 5 to 30 carbon atoms; the Z5 group is selected from hydrogen, -F, -Cl, -Br, hydroxyl, cyano, nitro, mercapto, amidino, hydrazine, hydrazone, an alkyl group having 1 to 10 carbon atoms, an alkenyl group having 2 to 10 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, an alkoxy group having 1 to 10 carbon atoms, a cycloalkyl group having 3 to 10 carbon atoms, a heterocycloalkyl group having 1 to 10 carbon atoms, a cycloalkenyl group having 3 to 10 carbon atoms, The invention can be any one of a heterocycloalkenyl group, an aryl group having 6 to 30 carbon atoms, an aryloxy group having 6 to 30 carbon atoms, an arylthio group having 6 to 30 carbon atoms, a heteroaryl group having 1 to 30 carbon atoms, a heteroaryloxy group having 1 to 30 carbon atoms, and a heteroarylthio group having 1 to 30 carbon atoms, and is preferably any one of an alkyl group having 1 to 10 carbon atoms and an alkoxy group having 1 to 10 carbon atoms; the number of substitutions of the Z5 group can be 1 to 2, and the substitution positions are not particularly limited; when the Z5 groups are multi-substituted, they can be the same or different.

[0015] In the technical solution of the present invention, the structural formula of the thiol-substituted heterocycle is A is any one of heteroaryl and fused heterocyclic ring having 5 to 30 carbon atoms; the heteroatom is any one or more of O, S and N; Z6 is hydrogen, -F, -Cl, -Br, hydroxyl, cyano, nitro, thiol, amidino, hydrazine, hydrazone, alkyl having 1 to 10 carbon atoms, alkenyl having 2 to 10 carbon atoms, alkynyl having 2 to 10 carbon atoms, alkoxy having 1 to 10 carbon atoms, cycloalkyl having 3 to 10 carbon atoms, heterocycloalkyl having 1 to 10 carbon atoms, or cycloalkyl having 3 to 10 carbon atoms. any one of a cycloalkenyl group having 1 to 10 carbon atoms, a heterocycloalkenyl group having 6 to 30 carbon atoms, an aryl group having 6 to 30 carbon atoms, an aryloxy group having 6 to 30 carbon atoms, an arylthio group having 6 to 30 carbon atoms, a heteroaryl group having 1 to 30 carbon atoms, a heteroaryloxy group having 1 to 30 carbon atoms, and a heteroarylthio group having 1 to 30 carbon atoms, preferably any one of a mercapto group and H; the number of substitutions of the Z6 group may be 1 to 2, and the substitution positions are not particularly limited; when the Z6 groups are multi-substituted, they may be the same or different.

[0016] In the technical solution of the present invention, the structural formula of the thiol-substituted aromatic ring is B is any one of an aryl group having 6 to 30 carbon atoms; hydrogen, -F, -Cl, -Br, hydroxyl, cyano, mercapto, amidino, hydrazine, hydrazone, an alkyl group having 1 to 10 carbon atoms, an alkenyl group having 2 to 10 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, an alkoxy group having 1 to 10 carbon atoms, a cycloalkyl group having 3 to 10 carbon atoms, a heterocycloalkyl group having 1 to 10 carbon atoms, a cycloalkenyl group having 3 to 10 carbon atoms, a heterocycloalkenyl group having 1 to 10 carbon atoms, a carbon atom Z7 is any one of an aryl group having 6 to 30 carbon atoms, an aryloxy group having 6 to 30 carbon atoms, an arylthio group having 6 to 30 carbon atoms, a heteroaryl group having 1 to 30 carbon atoms, a heteroaryloxy group having 1 to 30 carbon atoms, and a heteroarylthio group having 1 to 30 carbon atoms, and Z7 is preferably any one of an alkyl group having 1 to 10 carbon atoms and H; the number of substitutions of the Z7 group can be 1 to 2, and the substitution positions are not particularly limited; when the Z7 groups are multiple substitutions, they may be the same or different.

[0017] In certain specific embodiments, the cordycepin derivatives can be specifically listed as:

[0018]

[0019] In another aspect, the present invention provides a method for preparing the above cordycepin derivatives;

[0020] (I) R1 and R2 are alkoxycarbonyl, R3, R4 and R5 are H; the preparation method is shown in formula (II):

[0021]

[0022] The specific steps include:

[0023] S1: Cordycepin and tert-butyldimethylsilyl chloride (TBSCl) are reacted by substitution to prepare

[0024] S2: Prepared by amidation reaction

[0025] S3: Remove the protecting group on the product of step S2 by tetrabutylammonium fluoride (TBAF) to obtain

[0026] As a preferred embodiment, in step S1, the substitution reaction is carried out in an organic solvent; the organic solvent is selected from any one or more of CH2Cl2, tetrahydrofuran (THF) or N,N-dimethylformamide (DMF);

[0027] Preferably, in step S1, the substitution reaction is carried out under the action of an organic base; the organic base is selected from any one of imidazole, pyridine and Et3N;

[0028] In certain specific embodiments, Et3N is used as an organic base, and the substitution reaction further requires a catalyst; the catalyst is 4-dimethylaminopyridine (DMAP); the equivalent of the catalyst is 1% to 20%;

[0029] Preferably, the molar ratio of cordycepin to tert-butyldimethylchlorosilane is 1:2-15;

[0030] In certain specific embodiments, the concentration of cordycepin is 0.05 to 1 M;

[0031] Preferably, the substitution reaction does not require heating and the reaction time is 8 to 12 hours.

[0032] As a preferred embodiment, in step S2, the amidation reaction is carried out in an organic solvent; the organic solvent is selected from any one or more of N,N-dimethylformamide (DMF), dichloromethane (DCM) and tetrahydrofuran (THF);

[0033] Preferably, in step S2, the amidation reaction is carried out under the action of an organic base; the organic base is selected from any one of imidazole, pyridine and Et3N;

[0034] Preferably, in step S2, the amidation reaction is Carry out amidation reaction with R1-O-R2; The molar ratio with R1-O-R2 is 1:10-14;

[0035] Preferably, in step S2, the The molar ratio with the organic base is 1:0.1-2;

[0036] In certain specific embodiments, the The concentration is 0.01~0.5M;

[0037] Preferably, the amidation reaction is carried out at a temperature of 20 to 80° C. and for a time of 10 to 14 hours.

[0038] As a preferred embodiment, the reaction in step S3 is carried out in an organic solvent; the organic solvent is selected from any one of N,N-dimethylformamide (DMF), acetonitrile and tetrahydrofuran (THF);

[0039] Preferably, in step S3, the molar ratio of tetrabutylammonium fluoride (TBAF) to the product of step S2 is 1:40-60.

[0040] Preferably, the reaction temperature in step S3 is 20-80° C. and the reaction time is 3-5 h.

[0041] (ii) R3 is a substituted benzenesulfonyl group, and R1, R2, R4, and R5 are H; the preparation method comprises the following steps:

[0042] Cordycepin and sulfonyl chloride compounds are prepared by substitution reaction

[0043] As a preferred embodiment, the substitution reaction is carried out under the action of an organic base; the organic base is selected from any one of imidazole and pyridine;

[0044] Preferably, in the substitution reaction, the molar ratio of cordycepin to the sulfonyl chloride compound is 1:1-5;

[0045] Preferably, the temperature of the substitution reaction is 20 to 80°C;

[0046] In some specific embodiments, the concentration of cordycepin is 0.05-1M.

[0047] (III) R4 is a thiol-substituted heterocyclic ring or a thiol-substituted aromatic ring, and R1, R2, R3, and R5 are H; the preparation method is shown in formula (III):

[0048]

[0049] The specific steps include:

[0050] Step i: Synthesis of 2',3'-dehydroadenosine (3):

[0051] Will and 2-acetoxyisobutyryl bromide by substitution reaction

[0052]

[0053] Step ii: Prepare the product of step i by epoxidation reaction

[0054] Step iii: and R4H are prepared by substitution reaction

[0055] As a preferred embodiment, in step i, the substitution reaction is carried out in a solvent; the solvent is water and / or acetonitrile;

[0056] Preferably, the temperature of the substitution reaction is 20 to 80° C. and the time is 10 to 14 hours;

[0057] Preferably, The molar ratio of 2-acetoxyisobutyryl bromide to 2-acetoxyisobutyryl bromide is 1:3-5;

[0058] Preferably, the The concentration is 0.01~1M.

[0059] As a preferred embodiment, in step ii, the epoxidation reaction is carried out by subjecting the product of step i to the oxidation action of a strong base anion resin; the strong base anion resin is Amberlite IRA402 resin;

[0060] Preferably, the epoxidation reaction is carried out in an organic solvent; the organic solvent is selected from any one of methanol and ethanol;

[0061] In certain specific embodiments, the The concentration is 0.01~0.5M.

[0062] As a preferred embodiment, in step iii, the substitution reaction is carried out under the catalytic action of an organic base; the organic base is selected from any one of sodium methoxide and sodium tert-butoxide;

[0063] Preferably, in step iii, the substitution reaction is carried out in an organic solvent; the organic solvent is selected from any one of methanol and tert-butanol;

[0064] Preferably, in step iii, the The molar ratio of R4H to R4H is 1:1-3.

[0065] Preferably, in step iii, the temperature of the substitution reaction is 20 to 100°C;

[0066] In certain specific embodiments, the The concentration is 0.05~1M.

[0067] In another aspect, the present invention provides the use of the cordycepin derivatives in the preparation of antibacterial drugs, antioxidant drugs or cosmetics, and drugs for treating and / or preventing cancer.

[0068] In the technical solution of the present invention, the bacteria include Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Lactobacillus, Bifidobacterium and Candida albicans.

[0069] In the technical solution of the present invention, the cancer includes leukemia, breast cancer, gastric cancer and lung cancer.

[0070] Compared with the prior art, the present invention has the following advantages:

[0071] The present invention provides a cordycepin derivative to solve the technical problem of poor stability of cordycepin and its derivatives in the prior art. The cordycepin derivative uses a special group to modify the structure of cordycepin, which not only improves the stability of the cordycepin derivative, but also significantly improves the biological activities such as antibacterial, anti-inflammatory, antioxidant, and anti-tumor. In addition, the cordycepin derivative in the present invention has a certain selectivity for strains, especially for certain beneficial bacteria, in terms of antibacterial properties, which expands the application range of the cordycepin derivative. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 The product S3-1 obtained in Example 1 1 H NMR spectrum.

[0073] Figure 2 The product S2-4 obtained in Example 10 1 H NMR spectrum.

[0074] Figure 3 This is a graph showing the free radical scavenging rate of compound S3-2 in Example 13 as a function of concentration. DETAILED DESCRIPTION

[0075] The following embodiments are only some embodiments of the present invention, rather than all embodiments. Therefore, the detailed description in the embodiments of the present invention provided below is not intended to limit the scope of the present invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative work belong to the protection scope of the present invention.

[0076] In the present invention, unless otherwise specified, all equipment and raw materials can be purchased from the market or are commonly used in the industry. The methods in the following embodiments are all conventional methods in the art unless otherwise specified.

[0077] Preparation Example 1:

[0078]

[0079] The preparation method of compound 3 (2',3'-dehydrated adenosine) refers to non-patent document 1 "Hansske F, Robins MJ.Regiospecific and stereoselective conversion of ribonucleosides to 3'-deoxynucleosides.A high yield three-stage synthesis of cordycepin from adenosine.[J].Tetrahedron Letters,1985,26(36):4295-4298,DOI:10.1016 / S0040-4039(00)98716-1", and the specific process is as follows:

[0080] Adenosine (a, 267 mg) was suspended in "wet acetonitrile" (acetonitrile: water = 100: 1) (7.25 mL), and 2-acetoxyisobutyryl bromide (b, 0.60 mL) was added dropwise at 0°C. 20 mL of ultra-dry acetonitrile was added at room temperature, and the solution became clear after stirring for 1 h. After the raw material adenosine (a) was completely consumed, the reaction was quenched, extracted three times with ethyl acetate, and the organic layer was washed with a combination of Na2CO3 aqueous solution and water, dried over anhydrous Na2SO4, and concentrated by rotary evaporation to obtain a crude product (728.30 mg) as a yellow oil.

[0081] Activation of Amberlite IRA402(c) resin: Soak twice the volume of resin in 4% HCl for 4-8 hours, and wash with clean water until the pH is about 3-5. Rinse the resin further with 4 times the volume of 4% NaOH aqueous solution. Soak the resin in a 4% NaOH aqueous solution equal to the volume of the resin for 4-8 hours. Wash with clean water until the pH is neutral, and store in clean water for later use.

[0082] In a dry flask, add the crude product (728.30 mg) obtained in the previous step and 10 mL of methanol, and add newly activated resin (c) (7 mL). Stir at room temperature for 1 hour. Collect the reaction solution, rinse the resin with enough methanol and collect it, and concentrate it by rotary evaporation to obtain the crude product. Column chromatography gives 236.70 mg of white solid (3), with a yield of 95%.

[0083] Embodiment 1:

[0084] Synthesis steps of compound (S3-1):

[0085]

[0086] In a dry flask, sodium methoxide (86.40 mg) and 4-methylbenzenethiol (4,298.09 mg) were suspended in methanol (3.20 mL) under nitrogen protection and stirred at room temperature for 10 minutes; 2',3'-O-dehydrated adenosine (3,86.40 mg) obtained in Preparation Example 1 was added, and the mixture was refluxed at 75°C for 3 hours. After cooling, the crude product was concentrated by rotary evaporation and purified by column chromatography to obtain 80 mg of a cyan solid (Compound S3-1) with a yield of 67%.

[0087] Figure 1 Shown is compound S3-1 1 H NMR spectrum: 1 H NMR (400MHz, DMSO-d6) δ8.36(s,1H),8.15(s,1H),7.40(s,2H),7.36(d,J=8.2Hz,2H),7.14(d,J=8.0Hz,2H),6.09(d,J=5.8Hz, 1H),5.85-5.71(m,2H),4.78-4.62(m,1H),4.52(dt,J=8.1,2.8Hz,1H),3.92(t,J=8.3Hz,1H),3.83-3.67(m,2H),2.27(s,3H).

[0088] Embodiment 2:

[0089] Synthesis steps of compound (S3-2):

[0090]

[0091] In a dry flask, 2,5-dimercapto-1,3,4-thiadiazole (5, 66.10 mg) was dissolved in THF (5 mL), and sodium tert-butoxide was added to form a white suspension; the suspension was stirred for 10 minutes, 2',3'-O-dehydrated adenosine (3, 100 mg) obtained in Preparation Example 1 was added, and heated at 55°C for 6 h; after cooling, the crude product was concentrated by rotary evaporation and purified by column chromatography to obtain 46.50 mg of the product (compound S3-2) as a yellow oil with a yield of 29%.

[0092] Embodiment 3:

[0093] Synthesis steps of compound (S3-3):

[0094]

[0095] In a dry flask, sodium methoxide (207.92 mg) and 2-mercaptopyridine (6, 213.92 mg) were dissolved in methanol (8 mL), and the mixture was stirred for 10 minutes. 2', 3'-O-dehydrated adenosine (3, 200 mg) obtained in Preparation Example 1 was added, and sodium tert-butoxide was added to form a white suspension, which was refluxed at 75°C overnight. After cooling, the crude product was concentrated by rotary evaporation and purified by column chromatography to obtain 166 mg of a white solid compound (S3-3) with a yield of 57%.

[0096] Embodiment 4:

[0097] Synthesis steps of compound (S3-4):

[0098]

[0099] In a dry flask, sodium methoxide (312.14 mg) and 6-mercaptopurine (7,491.85 mg) were dissolved in methanol (3 mL), and the mixture was stirred for 10 minutes; 2',3'-O-dehydrated adenosine (3,240 mg) obtained in Preparation Example 1 was added, and the mixture was refluxed at 75°C overnight, and the crude product was concentrated by rotary evaporation after cooling. 367 mg of a cyan solid (S3-4) was obtained by column chromatography purification, with a yield of 95%.

[0100] Embodiment 5:

[0101] Synthesis steps of compound (S1-1):

[0102]

[0103] Synthesis of 6,6-bis(tert-butyloxycarbonyl)-3'-deoxyadenosine (S1-1):

[0104] In a dry flask, cordycepin (1,460 mg), imidazole (996.70 mg) and DMAP (22.36 mg) were added, and under nitrogen protection, DMF (16 mL) and TBSCl (1.10 g) were added at 0 ° C. Stir at room temperature for 15 hours. The reaction was quenched with saturated NH4Cl aqueous solution. It was extracted three times with EtOAc. The combined organic layers were washed with H2O and saturated NaCl aqueous solution, respectively, and concentrated by rotary evaporation to obtain a crude product. Purification by chromatography gave 605 mg of a white solid 2', 5'-bis-O-(tert-butyldimethylsilyl)-3'-deoxyadenosine (d) with a yield of 69%.

[0105] In a dry flask, add 2',5'-bis-O-(tert-butyldimethylsilyl)-3'-deoxyadenosine (S4, 600 mg, 1.25 mmol) and DMAP (30.56 mg), add THF (10 mL) at 0°C, and dropwise add Boc2O (3.60 mL). Stir at room temperature for 12 hours. Cool to room temperature, wash with saturated NaCl aqueous solution, dehydrate with anhydrous sodium sulfate, and concentrate by rotary evaporation to obtain a crude product. Purify by column chromatography to obtain 2',5'-bis-O-(tert-butyldimethylsilyl)-3'-deoxyadenosine (2, 786 mg) as a colorless oil with a yield of 92%.

[0106] In a dry flask, add 2',5'-bis-O-(tert-butyldimethylsilyl)-3'-deoxyadenosine (2,393 mg) to THF (3 mL), add TBAF (0.90 mL) dropwise at 0°C, and stir at room temperature for 4 h; purify by column chromatography to obtain 239 mg of white solid 6,6-bis(tert-butyloxycarbonyl)-3'-deoxyadenosine (S1-1), with a yield of 92%.

[0107] Embodiment 6:

[0108] Synthesis steps of compound (S1-2):

[0109]

[0110] In a dry flask, 6,6-bis(tert-butyloxycarbonyl)-3'-deoxyadenosine (S1-1, 80 mg) was dissolved in 3 mL of deionized water and placed at room temperature for more than 4 days. 47 mg of compound (S1-2) was purified by column chromatography with a yield of 76%.

[0111] Embodiment 7:

[0112] Synthesis steps of compound (S2-1):

[0113]

[0114] In a dry flask, cordycepin (1, 300 mg) was dissolved in pyridine (2 mL) at 0°C, TsCl (8, 409.76 mg) was added, and stirred at room temperature overnight; purified by column chromatography to obtain 106.50 mg of a light yellow solid (S2-1) with a yield of 16%.

[0115] Embodiment 8:

[0116] Synthesis steps of compound (S2-2):

[0117]

[0118] In a dry flask, cordycepin (1,200 mg) was dissolved in pyridine (2 mL) at 0°C, and then 4-methoxybenzenesulfonyl chloride (9,296.10 mg) was added and stirred at room temperature overnight; purified by column chromatography to obtain 108 mg of a light yellow solid (S2-2) with a yield of 32%.

[0119] Embodiment 9:

[0120] Synthesis steps of compound (S2-3):

[0121]

[0122] In a dry flask, add Boc2O (10,362 μL) to a solution of cordycepin (1,500 mg) and DMAP (48.60 mg) in THF (6 mL) at 0°C; heat the reaction to 65°C and stir for 12 hours; post-treatment: cool the reaction flask to room temperature, wash the organic phase with saturated saline solution, dehydrate with anhydrous sodium sulfate, and concentrate in vacuo; purify by column chromatography to obtain 46 mg of a white solid (S2-3) with a yield of 11%.

[0123] Embodiment 10:

[0124] Synthesis steps of compound (S2-4):

[0125]

[0126] In a dry flask, a solution of d (300 mg, 0.63 mmol) and TFA / H2O (4.60 mL, 1:1, 50 eq.) in THF (6.30 mL, 0.1 M) was stirred at 0°C for 1 hour; post-treatment: washed with saturated sodium bicarbonate solution, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated in vacuo; purified by column chromatography to give 111 mg of a white solid (S2-4) with a yield of 49%.

[0127] Characterization data: 1 H NMR (400MHz,CDCl3)δ8.30(s,1H),7.84(s,1H),6.36(s,2H),6.26(s,1H),5.61(d,J=5.8Hz,1H),5.04-4.94(m,1H),4.52-4.42(m,1H),3.97(dd,J=12.8,1.7Hz,1H),3.54(d,J=11.8Hz,1H),2.52(ddd,J=10.8,7.2,3.4Hz,1H),2.24-2.09(m,1H),0.76(s,9H),-0.15(s,3H),-0.27(s,3H).(See the specific spectrum for details) Figure 2 )

[0128] Example 11 Evaluation of antibacterial effect:

[0129] In this example, the two-fold dilution method was used to measure and evaluate the effect of the compound on the growth inhibition of the selected strain for 16 to 24 hours. The process was as follows: 100 μL of blank liquid culture medium was added to the 96-well plate, 100 μL of the test compound solution was added to the experimental group, the compound was diluted two-fold in sequence, and finally 10 μL of bacterial strain was added; only 100 μL of blank liquid culture medium and 10 μL of bacterial strain were added to the positive control group, and no test compound solution was added; only 110 μL of blank liquid culture medium was added to the negative control group; the above 96-well plate was placed in a water-insulated constant temperature incubator at 37°C and incubated for 16-24 hours, and then the OD was measured by an ELISA instrument. 600 All tests were performed three to five times independently. The test results are shown in Table 1-3.

[0130] Table 1: Antibacterial rate test data of 4800 μg / mL test compound solution

[0131]

[0132] Table 2: Antibacterial rate test data of 300 μg / mL test compound solution

[0133]

[0134] Table 3 Antibacterial performance of compounds at the maximum tested concentration

[0135]

[0136] In Table 3, “+” and “-” indicate the degree of inhibition: “+++” indicates that the bacterial solution is clear, “++” indicates slightly turbid, “+” indicates turbid, and “-” indicates very turbid.

[0137] Non-patent document 2 "Qu, Shuhao, et al." Design, synthesis, antibacterial / antitumoractivity and in vitro stability of novel cordycepin derivatives with unsaturated fatty acid chain. European Journal of Pharmaceutical Sciences 187 (2023): 106466" discloses cordycepin derivatives 1b and 1d, the structures of which are shown below:

[0138]

[0139] In vitro antibacterial activity of cordycepin derivatives 1b and 1d:

[0140]

[0141] As can be seen from the above table, the cordycepin derivatives S1-1, S2-3, and S2-4 in the present invention exhibit good antibacterial activity at the maximum test concentration, among which some cordycepin derivatives such as S1-1 have improved inhibitory activity on pathogenic bacteria while having a certain selectivity for beneficial bacteria. Cordycepin derivatives 1b and 1d in Document 1 need to reach 512 and 1024 μg / mL, respectively, to inhibit bacterial growth and cannot kill bacteria. In comparison, compounds S2-4, S2-3, S3-1, and S3-2 in this application all have obvious bactericidal effects at a concentration of 300 μg / mL, among which S2-4 has a bactericidal rate of 65% against Staphylococcus aureus, and compounds S1-1, S1-2, and 3 have stronger antibacterial effects at high concentrations.

[0142] In addition, the cordycepin derivatives in the present application have an antibacterial effect on certain pathogenic bacteria that is comparable to that of antibiotics, but are also relatively friendly to probiotics.

[0143] Example 12 Evaluation of the inhibitory effect on tumor cells:

[0144] This embodiment uses the CCK-8 method to detect cell viability, that is, cells growing in the logarithmic growth phase, aspirate the culture medium, trypsinize, add culture medium to terminate, gently blow, count, and inoculate 100 μL in a 96-well plate at a density of 2500 / well. After overnight attachment, discard the supernatant and add fresh culture medium containing a certain concentration of compound. Set two duplicate wells, add each concentration to the corresponding well, and culture in a 5% CO237°C incubator for 72 hours. Discard the supernatant and add a reaction solution mixed with fresh culture medium and CCK-8 in a ratio of 10 to 1. After incubation at 37°C for about 2 hours, use Thermo K3 to measure the light absorption value of 450nm (L1), refer to the wavelength of 630nm (L2), and calculate the relative cell activity by (L1-L2) value, and the effect of the compound on the cell can be obtained. The results are shown in Table 4. Cordycepin (1) and compound (S2-3) have certain inhibitory effects on cancer cells K562 (human chronic myeloid leukemia cells) and MGC-803 (gastric cancer cell line). The half-inhibitory concentrations of cordycepin (1) on K562 and MGC-803 are 37.32 μM and 16.10 μM, respectively, and the half-inhibitory concentrations of S2-3 on K562 and MGC-803 are 42.21 μM and 27.79 μM, respectively.

[0145] Table 4

[0146] Compound K562 MGC-803 Cordycepin 37.32μM 16.10μM S2-3 42.21μM 27.79μM

[0147] Example 13 Antioxidant Evaluation:

[0148] This embodiment tests the antioxidant properties of the above-mentioned derivatives. The specific process is: using 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) solution (0.3mmol / L, maximum absorption wavelength near 517nm) as the test substrate, it is divided into 4 groups, namely positive control group, negative control group, sample group and sample control group. The positive control group is 160μL Vc solution and 40μLDPH, the negative control group A is 160μL methanol and 40μL DPPH, and the sample group is 160μL compound solution and 40μL DPPH. Use a multifunctional microplate reader, set the wavelength to 517nm, and measure 3 times in parallel. The test results are shown in Table 5, where the free radical scavenging rate measured by compound (S3-2) varies with concentration as shown in the figure Figure 3 shown.

[0149] Table 5

[0150]

[0151]

[0152] Example 14 Stability Evaluation

[0153] This embodiment tests the stability of the above cordycepin derivatives, and the test process is as follows: weigh 0.007mmol of the drug to be tested, dissolve it in 40μL of DMSO, and dilute it to 10mL with a phosphate buffer solution of pH=7.4. Finally, take 0.5mL of the above solution in a centrifuge tube. Weigh 1mg of adenosine deaminase (64.8U) and dissolve it in 10mL of PBS solution of pH=7.4, take 1mL and dilute it to 10mL, take 50μL and add it to a centrifuge tube containing 0.5mL of the drug to be tested, and dilute it to 9mL. The experiment was carried out on a UV analyzer. In the presence of 3.5mU / mL adenosine deaminase, the absorption spectrum was scanned every 15min at a resolution of 2nm in the range of 230-300nm for 6-7h.

[0154] Table 6-8 shows the transformation of cordycepin (1), cordycepin derivative S1-1, and cordycepin derivative S2-3 under the action of adenosine deaminase (ADA enzyme) over time:

[0155] Table 6

[0156] time The remaining unconverted percentage 0min 100.00％ 15min 53.31％ 30min 34.80％ 45min 12.90％ 60min 2.72％ 75min 0.51％ 90min 0.00％

[0157] Table 7

[0158]

[0159]

[0160] Table 8

[0161] time The remaining unconverted percentage 0min 100.00％ 15min 79.10％ 30min 57.63％ 45min 50.28％ 60min 23.73％ 75min 9.04％ 90min 0.00％

[0162] It can be seen from the above table that the stability of the cordycepin derivatives S1-1 and S2-3 in the present application under the action of adenosine deaminase is improved to varying degrees compared with cordycepin.

[0163] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A cordycepin derivative, characterized in that: The general structural formula is shown in formula (I): In formula (I): R1 and R2 are each independently selected from any one of alkoxycarbonyl and H; R3 is selected from any one of an alkoxycarbonyl group, a silicon group, a substituted benzenesulfonyl group and H; R4 is selected from any one of a thiol-substituted heterocyclic ring, a thiol-substituted aromatic ring and H; R5 is any one of silicon group and H; R1 to R5 are not H at the same time.

2. The cordycepin derivative according to claim 1, characterized in that The structural formula of the alkoxycarbonyl group is The Z1 group is selected from any one of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms, a substituted or unsubstituted cycloalkyl group having 3 to 10 carbon atoms, a substituted or unsubstituted cycloalkenyl group having 3 to 10 carbon atoms, a substituted or unsubstituted aryl group having 6 to 30 carbon atoms, a substituted or unsubstituted aryloxy group having 6 to 30 carbon atoms, and a substituted or unsubstituted arylthio group having 6 to 30 carbon atoms, and is preferably any one of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms; Preferably, the silicon-based structure is The Z2 group, the Z3 group, and the Z4 group are each independently selected from any one of hydrogen, -F, -Cl, -Br, hydroxyl, cyano, nitro, amidino, hydrazine, hydrazone, alkyl having 1 to 10 carbon atoms, alkenyl having 2 to 10 carbon atoms, alkynyl having 2 to 10 carbon atoms, alkoxy having 1 to 10 carbon atoms, cycloalkyl having 3 to 10 carbon atoms, cycloalkenyl having 3 to 10 carbon atoms, aryl having 6 to 30 carbon atoms, aryloxy having 6 to 30 carbon atoms, and arylthio having 6 to 30 carbon atoms, and are preferably any one of alkyl having 1 to 10 carbon atoms; Preferably, the structural formula of the substituted benzenesulfonyl group is C is any one of an aryl group having 6 to 30 carbon atoms and a heteroaryl group having 5 to 30 carbon atoms; the Z5 group is selected from hydrogen, -F, -Cl, -Br, hydroxyl, cyano, nitro, thiol, amidino, hydrazine, hydrazone, an alkyl group having 1 to 10 carbon atoms, an alkenyl group having 2 to 10 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, an alkoxy group having 1 to 10 carbon atoms, a cycloalkyl group having 3 to 10 carbon atoms, a heterocycloalkyl group having 1 to 10 carbon atoms, a cycloalkenyl group having 3 to 10 carbon atoms, a carbon atom any one of heterocycloalkenyl having 1 to 10 carbon atoms, aryl having 6 to 30 carbon atoms, aryloxy having 6 to 30 carbon atoms, arylthio having 6 to 30 carbon atoms, heteroaryl having 1 to 30 carbon atoms, heteroaryloxy having 1 to 30 carbon atoms, heteroarylthio having 1 to 30 carbon atoms, preferably any one of alkyl having 1 to 10 carbon atoms and alkoxy having 1 to 10 carbon atoms; the number of substitutions of the Z5 group is 1 to 2; the Z5 groups are polysubstituted and are the same or different from each other; Preferably, the structural formula of the thiol-substituted heterocycle is A is any one of heteroaryl and fused heterocyclic ring having 5 to 30 carbon atoms; the heteroatom is any one or more of O, S and N; Z6 is hydrogen, -F, -Cl, -Br, hydroxyl, cyano, nitro, thiol, amidino, hydrazine, hydrazone, alkyl having 1 to 10 carbon atoms, alkenyl having 2 to 10 carbon atoms, alkynyl having 2 to 10 carbon atoms, alkoxy having 1 to 10 carbon atoms, cycloalkyl having 3 to 10 carbon atoms, heterocycloalkyl having 1 to 10 carbon atoms, Any one of a cycloalkenyl group having 3 to 10 atoms, a heterocycloalkenyl group having 1 to 10 carbon atoms, an aryl group having 6 to 30 carbon atoms, an aryloxy group having 6 to 30 carbon atoms, an arylthio group having 6 to 30 carbon atoms, a heteroaryl group having 1 to 30 carbon atoms, a heteroaryloxy group having 1 to 30 carbon atoms, and a heteroarylthio group having 1 to 30 carbon atoms, preferably any one of a mercapto group and H; the number of substitutions of the Z6 group is 1 to 2; the Z6 groups are polysubstituted and are the same or different from each other; Preferably, the structural formula of the thiol-substituted aromatic ring is B is any one of an aryl group having 6 to 30 carbon atoms; hydrogen, -F, -Cl, -Br, hydroxyl, cyano, mercapto, amidino, hydrazine, hydrazone, an alkyl group having 1 to 10 carbon atoms, an alkenyl group having 2 to 10 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, an alkoxy group having 1 to 10 carbon atoms, a cycloalkyl group having 3 to 10 carbon atoms, a heterocycloalkyl group having 1 to 10 carbon atoms, a cycloalkenyl group having 3 to 10 carbon atoms, Any one of heterocycloalkenyl, aryl having 6 to 30 carbon atoms, aryloxy having 6 to 30 carbon atoms, arylthio having 6 to 30 carbon atoms, heteroaryl having 1 to 30 carbon atoms, heteroaryloxy having 1 to 30 carbon atoms, heteroarylthio having 1 to 30 carbon atoms, preferably Z7 is any one of alkyl having 1 to 10 carbon atoms and H; the number of substitutions of the Z7 group is 1 to 2; the Z7 groups are polysubstituted and are the same or different from each other.

3. The method for preparing a cordycepin derivative according to claim 1 or 2, characterized in that: The preparation method is method (1), method (2) or method (3); Method (I): R1 and R2 are alkoxycarbonyl, R3, R4 and R5 are H, and the preparation method is shown in formula (II): The specific steps include: S1: Cordycepin and tert-butyldimethylsilyl chloride (TBSCl) are reacted by substitution to prepare S2: Prepared by amidation reaction S3: Remove the protecting group on the product of step S2 by tetrabutylammonium fluoride (TBAF) to obtain Method (II): R3 is a substituted benzenesulfonyl group, R1, R2, R4, and R5 are H, and the preparation method comprises the following steps: Cordycepin and sulfonyl chloride compounds are prepared by substitution reaction Method (III): R4 is a thiol-substituted heterocyclic ring or a thiol-substituted aromatic ring, R1, R2, R3, and R5 are H, and the preparation method is shown in formula (III): The specific steps include: Step i: Synthesis of 2',3'-dehydroadenosine (3): Will and 2-acetoxyisobutyryl bromide by substitution reaction Step ii: Prepare the product of step i by epoxidation reaction Step iii: and R4H are prepared by substitution reaction 4. The preparation method according to claim 3, characterized in that: In the method (I): In step S1, the substitution reaction is carried out in an organic solvent; the organic solvent is selected from any one or more of CH2Cl2, tetrahydrofuran or N,N-dimethylformamide; Preferably, in step S1, the substitution reaction is carried out under the action of an organic base; the organic base is selected from any one of imidazole, pyridine and Et3N; Preferably, the molar ratio of cordycepin to tert-butyldimethylchlorosilane is 1:2-15; Preferably, the reaction time of the substitution reaction is 8 to 12 hours.

5. The preparation method according to claim 3, characterized in that: In the method (I): In step S2, the amidation reaction is carried out in an organic solvent; the organic solvent is selected from any one or more of N,N-dimethylformamide, dichloromethane and tetrahydrofuran; Preferably, in step S2, the amidation reaction is carried out under the action of an organic base; the organic base is selected from any one of imidazole, pyridine and Et3N; Preferably, in step S2, the amidation reaction is Carry out amidation reaction with R1-O-R2; The molar ratio with R1-O-R2 is 1:10-14; Preferably, in step S2, the The molar ratio with the organic base is 1:0.1-2; Preferably, the amidation reaction is carried out at a temperature of 20 to 80° C. and for a time of 10 to 14 hours.

6. The preparation method according to claim 3, characterized in that: In the method (I): The reaction in step S3 is carried out in an organic solvent; the organic solvent is selected from any one of N,N-dimethylformamide, acetonitrile and tetrahydrofuran; Preferably, in step S3, the molar ratio of tetrabutylammonium fluoride to the product of step S2 is 1:40-60; Preferably, the reaction temperature in step S3 is 20-80° C. and the reaction time is 3-5 h.

7. The preparation method according to claim 3, characterized in that: In the method (ii), the substitution reaction is carried out under the action of an organic base; the organic base is selected from any one of imidazole and pyridine; Preferably, in the substitution reaction, the molar ratio of cordycepin to the sulfonyl chloride compound is 1:1-5; Preferably, the temperature of the substitution reaction is 20-80°C.

8. The preparation method according to claim 3, characterized in that: In the method (ii): In step i, the substitution reaction is carried out in a solvent; the solvent is water and / or acetonitrile; Preferably, the temperature of the substitution reaction is 20 to 80° C. and the time is 10 to 14 hours; Preferably, The molar ratio of 2-acetoxyisobutyryl bromide to 2-acetoxyisobutyryl bromide is 1:3-5; Preferably, the The concentration is 0.01~1M.

9. The preparation method according to claim 3, characterized in that: In the method (ii): In step ii, the epoxidation reaction is carried out by subjecting the product of step i to the oxidation action of a strong base anion resin; the strong base anion resin is Amberlite IRA402 resin; Preferably, the epoxidation reaction is carried out in an organic solvent; the organic solvent is selected from any one of methanol and ethanol; Preferably, in step iii, the substitution reaction is carried out under the catalytic action of an organic base; the organic base is selected from any one of sodium methoxide and sodium tert-butoxide; Preferably, in step iii, the substitution reaction is carried out in an organic solvent; the organic solvent is selected from any one of methanol and tert-butanol; Preferably, in step iii, the The molar ratio of R4H is 1:1-3; Preferably, in step iii, the temperature of the substitution reaction is 20-100°C.

10. Use of the cordycepin derivative according to claim 1 or 2 in the preparation of antibacterial drugs, antioxidant drugs or cosmetics, and drugs for treating and / or preventing cancer; Preferably, the bacteria include Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Lactobacillus, Bifidobacterium and Candida albicans; Preferably, the cancer includes leukemia, breast cancer, gastric cancer and lung cancer.

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