CN109111445B - Synthesis method and application of 5 '-furoyl ester-3' -deoxyadenosine - Google Patents
Synthesis method and application of 5 '-furoyl ester-3' -deoxyadenosine Download PDFInfo
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- C07D473/00—Heterocyclic compounds containing purine ring systems
- C07D473/26—Heterocyclic compounds containing purine ring systems with an oxygen, sulphur, or nitrogen atom directly attached in position 2 or 6, but not in both
- C07D473/32—Nitrogen atom
- C07D473/34—Nitrogen atom attached in position 6, e.g. adenine
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Abstract
A synthesis method and application of 5 '-furoyl ester-3' -deoxyadenosine belong to the technical field of synthesis of cordycepin analogues. The preparation method comprises the steps of weighing cordycepin bulk drug and anhydrous pyridine respectively, adding the weighed cordycepin bulk drug into the anhydrous pyridine, stirring and dissolving to obtain a mixed solution, slowly dropwise adding furoyl chloride with certain mass under the conditions of nitrogen protection and magnetic heating and stirring for reaction, monitoring the reaction process by using a thin-layer chromatography, carrying out reduced pressure distillation after the reaction to obtain a paste product, dissolving by using distilled water, extracting by using ethyl acetate for 3 times, combining organic phase solutions, carrying out reduced pressure distillation to remove an organic solvent, mixing the obtained solid powder mixture with column chromatography silica gel, and carrying out column chromatography by using a dry-method sample loading method to obtain the 5 '-furoyl ester-3' -deoxyadenosine. The method is used for synthesizing the analog of cordycepin.
Description
Technical Field
The invention belongs to the technical field of synthesis of cordycepin analogues; in particular to a synthesis method and application of 5 '-furoyl ester-3' -deoxyadenosine.
Background
Cordycepin is an analogue of adenosine, also called 3' -deoxyadenosine, and is separated from culture filtrate of traditional Chinese medicine Cordyceps militaris (Cordyceps) in China as early as 1951 by Cunningham and the like in Germany, is the first nucleoside antibiotic separated from fungi, and has various pharmacological effects of resisting tumors and leukemia, resisting bacteria, regulating immunity, eliminating free radicals in vivo and the like. At present, the research of cordycepin has become an extremely active field in medicinal chemistry.
Tumor diseases are one of the diseases with high incidence at present, and in recent years, the research on the pharmacological action mechanism, especially the anti-tumor action mechanism, of cordycepin (3' -deoxyadenosine), which is the active ingredient in cordyceps sinensis, has become one of the extremely active research fields. Cordycepin is reported to have growth inhibition effect, so that cordycepin is widely researched as a potential anti-tumor drug.
Disclosure of Invention
The invention aims to provide a synthesis method and application of 5 '-furoyl ester-3' -deoxyadenosine.
The invention is realized by the following technical scheme:
a method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine comprises the following steps:
step 3, carrying out reduced pressure distillation on the reaction liquid obtained in the step 2 to obtain a paste product for later use;
step 4, dissolving the paste product obtained in the step 3 with distilled water, extracting for 3 times with ethyl acetate, and combining organic phase solutions for later use;
and 5, distilling the organic phase solution obtained in the step 4 under reduced pressure to remove the organic solvent to obtain a solid powder mixture, uniformly mixing the solid powder mixture with column chromatography silica gel, and carrying out column chromatography by a dry-method sample loading method to obtain the 5 '-furoyl ester-3' -deoxyadenosine.
The synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine comprises the following steps that in the step 1, the material-liquid ratio of cordycepin bulk drug to anhydrous pyridine is 1 g: 45-55 ml.
According to the synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine, the mass ratio of furoyl chloride in the step 2 to cordycepin raw material medicine in the step 1 is 0.55-0.65: 1, heating in a water bath mode, wherein the heating temperature is 68-72 ℃, and the reaction time is 4.8-5.2 h.
According to the synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine, in the step 2, a thin-layer chromatography solvent is a mixed solvent of methanol and dichloromethane, wherein the volume ratio of the methanol to the dichloromethane is 1:10, a thin-layer plate is observed under an ultraviolet analyzer, the raw material point disappears, and the reaction is finished.
According to the synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine, in the step 3, a rotary evaporator is used for carrying out reduced pressure distillation at the temperature of 80 ℃, and the distillation time is 5-10 min.
The synthesis method of 5 '-furoyl ester-3' -deoxyadenosine comprises the following steps of adding distilled water in the step 4 at the temperature of 60 ℃, wherein the volume ratio of the added distilled water to ethyl acetate is 10 ml: 25-35 ml.
The invention relates to a synthesis method of 5 '-furoyl ester-3' -deoxyadenosine, in step 5, a developing agent for column chromatography is a mixed solvent of methanol and dichloromethane, and the volume ratio of the methanol to the dichloromethane is 1: and 15, putting the mixture into a silica gel column which is a2 multiplied by 75cm glass column, and performing 200-300-mesh column chromatography on silica gel.
The 5 '-furoyl ester-3' -deoxyadenosine prepared by the synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine is applied to preparation of a medicament for inhibiting cancers, wherein the cancers comprise liver cancers.
The synthesis reaction formula of the 5 '-furoyl ester-3' -deoxyadenosine is shown in the reaction formula (1):
the synthetic method of the 5 '-furoyl ester-3' -deoxyadenosine comprises the following steps of:
the invention has the following beneficial effects:
the yield of the synthetic method of the 5 '-furoyl ester-3' -deoxyadenosine is more than 50%.
The synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine, disclosed by the invention, is used for synthesizing the 5 '-furoyl ester-3' -deoxyadenosine of the cordycepin analogue, and the characterization and confirmation of the structure of the 5 '-furoyl ester-3' -deoxyadenosine are carried out through infrared spectroscopy, nuclear magnetic resonance hydrogen spectrum, carbon spectrum and mass spectrum; and (3) measuring the in vitro anti-tumor activity of the compound obtained by synthesis by using an MTT colorimetric method.
The synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine provided by the invention is used for researching the in vitro anti-tumor activity, provides a basis for the deep research of the anti-tumor effect of the cordycepin structural analogue and the exploration of the prospect of the cordycepin structural analogue in the aspect of tumor treatment application, and is expected to provide a new thought for the drug development of the cordycepin structural analogue and lay a theoretical basis for clinical application.
The 5 '-furoyl ester-3' -deoxyadenosine synthesized by the synthesis method of the 5 '-furoyl ester-3' -deoxyadenosine can improve the drug effect of cordycepin.
The 5 '-furoyl ester-3' -deoxyadenosine synthesized by the invention improves the inhibition effect of cordycepin on liver cancer cell HepG-2. 3' -deoxyadenosine as a control, IC50The value was 19.18 ug/ml. IC of 5 '-furoyl ester-3' -deoxyadenosine50The value was 8.891 ug/ml.
Drawings
FIG. 1 is an IR spectrum of 5 '-furoyl ester-3' -deoxyadenosine prepared by a method according to an embodiment;
FIG. 2 is an infrared absorption spectrum of 3' -deoxyadenosine;
FIG. 3 is a nuclear magnetic hydrogen spectrum of 5 '-furoyl ester-3' -deoxyadenosine prepared by a method according to an embodiment;
FIG. 4 is a nuclear magnetic carbon spectrum of 5 '-furoyl ester-3' -deoxyadenosine prepared by a method according to an embodiment;
FIG. 5 is a mass spectrum of 5 '-furoyl ester-3' -deoxyadenosine prepared by a method according to an embodiment;
FIG. 6 is a MTT assay inhibition curve for 5 '-furoyl ester-3' -deoxyadenosine prepared by a method of an embodiment.
Detailed Description
The first embodiment is as follows:
a method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine comprises the following steps:
step 3, carrying out reduced pressure distillation on the reaction liquid obtained in the step 2 to obtain a paste product for later use;
step 4, dissolving the paste product obtained in the step 3 with distilled water, extracting for 3 times with ethyl acetate, and combining organic phase solutions for later use;
and 5, distilling the organic phase solution obtained in the step 4 under reduced pressure to remove the organic solvent to obtain a solid powder mixture, uniformly mixing the solid powder mixture with column chromatography silica gel, and carrying out column chromatography by a dry-method sample loading method to obtain the 5 '-furoyl ester-3' -deoxyadenosine.
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, in the step 1, the ratio of the cordycepin bulk drug to the anhydrous pyridine is 1 g: 50 ml.
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, the mass ratio of furoyl chloride in the step 2 to cordycepin bulk drug in the step 1 is 0.6: 1, heating in a water bath mode, wherein the heating temperature is 70 ℃, and the reaction time is 5 hours.
In the synthesis method of 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, the thin-layer chromatography solvent in the step 2 is a mixed solvent of methanol and dichloromethane, wherein the volume ratio of the methanol to the dichloromethane is 1:10, and when the thin-layer plate is observed under an ultraviolet analyzer, the raw material point disappears and the reaction is finished.
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, in the step 3, a rotary evaporator is used for carrying out reduced pressure distillation at 80 ℃, and the distillation time is 5-10 min.
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, the temperature of the distilled water added in the step 4 is 60 ℃, and the volume ratio of the added distilled water to the ethyl acetate is 10 ml: 30 ml.
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, in step 5, a developing solvent for column chromatography is a mixed solvent of methanol and dichloromethane, and the volume ratio of the methanol to the dichloromethane is 1: 15, the silica gel column is a2 multiplied by 75cm glass column, and the silica gel is 200-mesh column chromatography silica gel.
According to the synthesis method of 5 '-furoyl ester-3' -deoxyadenosine, when the amount of cordycepin raw material drug is 1g, 1.4369g of white solid powder obtained in the step 5 is obtained, and 0.695g of 5 '-furoyl ester-3' -deoxyadenosine is finally obtained, 1.38g of 5 '-furoyl ester-3' -deoxyadenosine is theoretically generated, and the yield is 50.4%.
In the method for synthesizing 5' -furoyl ester-3 ' -deoxyadenosine according to the embodiment, an infrared spectrum test is performed on the synthesized 5' -furoyl ester-3 ' -deoxyadenosine, as shown in fig. 1, fig. 2 is an infrared absorption spectrum of 3' -deoxyadenosine as a comparison spectrum, and as can be seen from the peak value comparison of fig. 1 and fig. 2, the characteristic peak value of 5' -furoyl ester-3 ' -deoxyadenosine in fig. 1 is 3486cm-1、3159cm-1、2934cm-1、2857cm-1、1736cm-1、1713cm-1、1662cm-1、1595cm-1、1479cm-1、1393cm-1、963cm-1、729，636cm-1(ii) a The characteristic peak value of 3' -deoxyadenosine (cordycepin) in figure 2 is 3337cm-1、3141cm-1、2934cm-1、1671cm-1、1617cm-1、1479cm-1、1342cm-1、1304cm-1、829cm-1、633cm-1(ii) a Wherein the length of the groove is 3141cm-1Is a characteristic absorption peak of 5-hydroxyl, 3337cm-1Is the flexible vibration absorption peak of N-H at the 6 position, 1341cm-1The hydroxyl absorption peak of the 5-position primary alcohol. FIG. 1 is 1341cmcm in FIG. 1 compared with FIG. 2-1The absorption peak disappeared. The product is 1736cmcm-1Shows a strong absorption peak which is a characteristic absorption peak of the ester carbonyl group, 1194cmcm-1The broad peak is the characteristic absorption peak of C ═ O on the ester carbonyl.
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, nuclear magnetic hydrogen spectrum analysis is performed on the synthesized 5 '-furoyl ester-3' -deoxyadenosine, a solvent DMSO is tested, the frequency is 10000Hz, and the test result is as followsAs shown in fig. 3, 13 signal peaks are obtained, which are:1HNMR(500MHz，DMSO-d6)8.26(1H，s，2-H)，8.13(1H，d，8-H)，7.98-6.70(3H，d，5’-CH＝)，5.93(1H，d，1’-H)，5.77(1H，d，1’-H)4.69-4.24(4H，ddtd，2’-OH，4’-H，5’-CH2)，2.06(2H，ddd，3’-CH2)。
in the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, nuclear magnetic carbon spectrum analysis is performed on the synthesized 5 '-furoyl ester-3' -deoxyadenosine to test a solvent DMSO at a test frequency of 29761.904Hz, and a test result is shown in fig. 4, so that 15 signal peaks are obtained, wherein the signal peaks are respectively: 13CNMR (126MHz, DMSO-d6)158.16(1C, -COO), 156.52(1C, 6-C), 153.10(1C, 2-C), 149.47(1C, 4-C), 148.27-143.97(2C, 5' -CH ═ CH), 139.31(1C, 8-C), 119.44(1C, 5-C), 119.15-112.85(2C, 5' -CH ═ CH), 91.22(1C, 1 ' -C), 77.87(1C, 4 ' -C), 74.81(1C, 2 ' -C), 65.95(1C, 5' -C), 35.17(1C, 3' -C).
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, mass spectrometry is performed on the synthesized 5 '-furoyl ester-3' -deoxyadenosine, and the test result is shown in fig. 5, wherein the mass spectrometry test result shows that the compound has a relative molecular weight of 346.1154 and a predicted molecular weight of 345, and the test result corresponds to the prediction result.
The second embodiment is as follows:
one specific embodiment is the application of 5 '-furoyl ester-3' -deoxyadenosine prepared by a synthesis method in preparing a medicament for inhibiting cancers, wherein the cancers comprise liver cancers.
In the embodiment, the 5 '-furoyl ester-3' -deoxyadenosine is applied to the preparation of the cancer inhibiting drug, the 5 '-furoyl ester-3' -deoxyadenosine is used for cell proliferation detection, and the detection method adopts an MTT (methyl thiazolyl tetrazolium) detection method:
the first step is cell recovery: taking out the freezing tube filled with HepG2 cells from the liquid nitrogen tank, quickly putting the freezing tube into a preheated water bath kettle, quickly thawing at 37 ℃, and continuously shaking to quickly thaw the liquid in the tube. After about 1min, the liquid in the freezing storage tube is completely dissolved, and the alcohol cotton is taken outWiping the outer wall of the cryopreservation tube with a ball, placing the cryopreservation tube into an ultra-clean workbench, sucking out cell suspension by using a suction pipe, injecting the cell suspension into a centrifuge tube, adding 10 times of culture solution, mixing, and centrifuging at 1000rpm for 5 min. The supernatant was aspirated off, 10mL of culture medium was added to the tube, and the tube was aspirated to obtain a cell suspension. Counting cells, adjusting cell concentration to 3 × 104cell/mL. Subpackaging the cell suspension into culture bottles, and placing the culture bottles in a temperature of 37 ℃ and 5% CO2Culturing in an incubator, and replacing the culture solution once the next day.
The second step is subculture of cells: human hepatoma cell line HepG2Culturing in RPMI-1640 culture medium containing 10% calf serum at 37 deg.C and 5% C02In a saturated humidity incubator, when the adherent growth of the cells is 80% -90% fused, washing the cells twice by PBS liquid, digesting the cells by 0.25% trypsin, observing under a microscope until most of the cells are rounded, removing digestive juice, adding 5mL of 10% calf serum RPMI-1640 culture solution into a culture bottle, repeatedly and fully blowing the bottle wall by a straw to prepare single cell suspension, adding a proper amount of 10% calf serum-containing RPMI-1640 culture solution according to the principle of one bottle transferring two bottles or three bottles, uniformly mixing, respectively filling into each bottle, placing at 37 ℃, and placing at 5% C02Culturing in a saturated humidity incubator. Counting under the mirror, adjusting the cell concentration to 3X 104one/mL.
The third step is solution preparation: PBS: accurately weighing KCL0.4g, NaCL16g, Na2HP 04.12 H205.76g, KH2P040.4g and ddH20, dissolving to constant volume of 2L, adjusting the pH of the solution to 7.2-7.4 with HCL, filtering with 0.22 μm sterile filter membrane, sterilizing under high pressure, and storing in refrigerator at 4 deg.C. MTT: MTT50mg was weighed, dissolved by ultrasonic in 10ml PBS under dark, filtered through 0.22 μm filter membrane for sterilization, and stored at 4 ℃ under dark.
The fourth step is IC50 determination: the cells were cultured at 5X 105The inoculum size of each/mL is inoculated in a 96-well culture plate, and 5% CO is added2Culturing in 37 deg.C incubator for 24 hr, adding 10 μ L of each drug with different concentrations into each well to make final concentration of the primarily screened drugs 103.10210 μ g/mL, and incubation was continued for 24h, while the final drug concentration screened was 100.200.400.600.800 μ g/mL, respectively, with 5 parallel wells per well. The zeroing well was not filled with cell suspension, but with 10% fetal bovine serum RPMI-1640200 μ L, culturing for 48 hr, adding 50 μ LMTT (thiazole blue) respectively, incubating for 4 hr, discarding the culture medium, adding 150 μ LDMSO, shaking on a plate shaker, reading the plate under enzyme labeling instrument 490nm, and calculating IC with analysis software SPSS according to the measured absorbance value50The value is obtained.
The fifth step is the determination of the cell inhibition rate: taking HepG2 cells in logarithmic growth phase, centrifuging at 1000rpm for 10min, and adjusting the concentration of the precipitated cells to 1X 10 by using RPMI-1640 culture solution (containing 10% fetal bovine serum)5cell·mL-1After the cell suspension, the cells were seeded in a 96-well culture plate. Add 100. mu.L (1X 10) of cell suspension per well4cell), then respectively adding N-benzoyl cordycepin, N-thiophene amide cordycepin, N-furan amide cordycepin and cordycepin with different concentrations to 200 mu g/mL-1And 100. mu.L each of the drug-free culture medium, 4 wells in parallel, to make a total volume of 200. mu.L. The zeroing wells did not contain HepG2 cell suspension, but only 10% fetal bovine serum RPMI-1640200. mu.L. Mixing, adding 5% CO at 37 deg.C2After culturing for 48 hours under 95% humidity conditions, 5 mg/mL of the solution was added to each well-120. mu.L of MTTPBS solution was cultured under the same conditions for 4 hours, and the culture was terminated. Centrifuging at 1000rpm for 5min, removing the culture solution in the culture plate holes, adding 150 mu LDMSO into each hole, shaking for 10min to fully dissolve the formed formazan particles, and detecting the light absorption value by an enzyme-labeling instrument. The measurement wavelength was chosen to be 490 nm. The cell growth inhibition rate was calculated according to the following formula, and the test results are shown in table 1:
cell growth inhibition (%) (1-experimental well OD 570/control well OD570) × 100% (3)
TABLE 1MTT test COMPARATIVE TABLE
As can be seen from Table 1, 5 '-furoyl ester-3' -deoxyadenosine has an inhibitory effect (P <0.01) on liver cancer cell HepG-2, the inhibition rate is 50%, the inhibitory effect is shown in figure 6, as can be seen from figure 6, the inhibitory effect of 5 '-furoyl ester-3' -deoxyadenosine on liver cancer cell HepG-2 is stronger and stronger along with the increase of the administration concentration, and when the concentration reaches 8.891ug/ml, the inhibitory effect reaches half inhibition of cell growth.
The third concrete implementation mode:
a method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine comprises the following steps:
step 3, carrying out reduced pressure distillation on the reaction liquid obtained in the step 2 to obtain a paste product for later use;
step 4, dissolving the paste product obtained in the step 3 with distilled water, extracting for 3 times with ethyl acetate, and combining organic phase solutions for later use;
and 5, distilling the organic phase solution obtained in the step 4 under reduced pressure to remove the organic solvent to obtain a solid powder mixture, uniformly mixing the solid powder mixture with column chromatography silica gel, and carrying out column chromatography by a dry-method sample loading method to obtain the 5 '-furoyl ester-3' -deoxyadenosine.
The method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment has a yield of 50% or more.
In the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the embodiment, the 5 '-furoyl ester-3' -deoxyadenosine is synthesized as a cordycepin analogue, and the structure of the 5 '-furoyl ester-3' -deoxyadenosine is characterized and confirmed by an infrared spectrum, a nuclear magnetic resonance hydrogen spectrum, a carbon spectrum and a mass spectrum; and (3) measuring the in vitro anti-tumor activity of the compound obtained by synthesis by using an MTT colorimetric method.
The synthesis method of 5 '-furoyl ester-3' -deoxyadenosine in the embodiment is used for researching the in vitro antitumor activity, provides a basis for the deep research of the antitumor effect of the cordycepin structural analogue and the exploration of the prospect of the cordycepin structural analogue in the aspect of tumor treatment application, and is expected to provide a new idea for the drug development of the cordycepin structural analogue and lay a theoretical basis for clinical application.
The fourth concrete implementation mode:
according to the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the third embodiment, the ratio of the cordycepin bulk drug to the anhydrous pyridine in the step 1 is 1 g: 45-55 ml.
The fifth concrete implementation mode:
according to the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine in the third embodiment, the mass ratio of furoyl chloride in the step 2 to cordycepin bulk drug in the step 1 is 0.55-0.65: 1, heating in a water bath mode, wherein the heating temperature is 68-72 ℃, and the reaction time is 4.8-5.2 h.
The sixth specific implementation mode:
according to the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to the third embodiment, the thin-layer chromatography solvent in the step 2 is a mixed solvent of methanol and dichloromethane, wherein the volume ratio of the methanol to the dichloromethane is 1:10, and when the thin-layer plate is observed under an ultraviolet analyzer, the raw material point disappears and the reaction is finished.
The seventh embodiment:
according to the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine in the third embodiment, in the step 3, the rotary evaporator is used for carrying out reduced pressure distillation at 80 ℃, and the distillation time is 5-10 min.
The specific implementation mode is eight:
according to the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine of the third embodiment, the temperature of the distilled water added in the step 4 is 60 ℃, and the volume ratio of the added distilled water to the ethyl acetate is 10 ml: 25-35 ml.
The specific implementation method nine:
according to the method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine of the third embodiment, the developing solvent for column chromatography in the step 5 is a mixed solvent of methanol and dichloromethane, and the volume ratio of the methanol to the dichloromethane is 1: and 15, putting the mixture into a silica gel column which is a2 multiplied by 75cm glass column, and performing 200-300-mesh column chromatography on silica gel.
The detailed implementation mode is ten:
the application of the 5 '-furoyl ester-3' -deoxyadenosine prepared by the method for synthesizing the 5 '-furoyl ester-3' -deoxyadenosine according to one of the third embodiment to the ninth embodiment in preparing a medicament for inhibiting cancers, wherein the cancers comprise liver cancers.
Claims (4)
1. A method for synthesizing 5 '-furoyl ester-3' -deoxyadenosine is characterized in that: the method comprises the following steps:
step 1, weighing cordycepin bulk drug and anhydrous pyridine respectively according to a certain feed-liquid ratio, adding the weighed cordycepin bulk drug into the anhydrous pyridine, stirring and dissolving to obtain a mixed solution for later use;
step 2, slowly dripping furoyl chloride with certain mass into the mixed solution obtained in the step 1 under the conditions of nitrogen protection and magnetic heating and stirring for reaction, monitoring the reaction process by using thin-layer chromatography, and obtaining reaction liquid for later use after the reaction;
step 3, carrying out reduced pressure distillation on the reaction liquid obtained in the step 2 to obtain a paste product for later use;
step 4, dissolving the paste product obtained in the step 3 with distilled water, extracting for 3 times with ethyl acetate, and combining organic phase solutions for later use;
step 5, distilling the organic phase solution obtained in the step 4 under reduced pressure to remove the organic solvent to obtain a solid powder mixture, uniformly mixing the solid powder mixture with column chromatography silica gel, and carrying out column chromatography by a dry-method sample loading method to obtain 5 '-furoyl ester-3' -deoxyadenosine;
the mass ratio of the furoyl chloride in the step 2 to the cordycepin raw material medicine in the step 1 is 0.55-0.65: 1, heating in a water bath mode, wherein the heating temperature is 68-72 ℃, and the reaction time is 4.8-5.2 h;
in the step 1, the ratio of the cordycepin bulk drug to the anhydrous pyridine is 1 g: 45-55 ml;
in the step 2, the thin-layer chromatography solvent is a mixed solvent of methanol and dichloromethane, wherein the volume ratio of the methanol to the dichloromethane is 1:10, and when the thin-layer plate is observed under an ultraviolet analyzer, the raw material point disappears and the reaction is finished;
the temperature of the distilled water added in the step 4 is 60 ℃, and the volume ratio of the added distilled water to the ethyl acetate is 10 ml: 25-35 ml.
2. The method of synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to claim 1, wherein: and 3, carrying out reduced pressure distillation at 80 ℃ by using a rotary evaporator, wherein the distillation time is 5-10 min.
3. The method of synthesizing 5 '-furoyl ester-3' -deoxyadenosine according to claim 1, wherein: step 5, the developing agent for column chromatography is a mixed solvent of methanol and dichloromethane, and the volume ratio of the methanol to the dichloromethane is 1: and 15, putting the mixture into a silica gel column which is a2 multiplied by 75cm glass column, and performing 200-300-mesh column chromatography on silica gel.
4. Use of 5 '-furoyl ester-3' -deoxyadenosine prepared by the synthetic method of claims 1-3 in the preparation of a medicament for inhibiting cancer, characterized in that: the cancer is liver cancer.
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