CN100355901C - Synthesis method of 5-fluorocytidine - Google Patents
Synthesis method of 5-fluorocytidine Download PDFInfo
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- CN100355901C CN100355901C CNB200510029428XA CN200510029428A CN100355901C CN 100355901 C CN100355901 C CN 100355901C CN B200510029428X A CNB200510029428X A CN B200510029428XA CN 200510029428 A CN200510029428 A CN 200510029428A CN 100355901 C CN100355901 C CN 100355901C
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- flurocytosine
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- damping fluid
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- thymidine
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Abstract
The present invention provides a method for synthesizing 5-fluorocytidine. In the method, 5-fluorocytidine is synthesized from thymidine and 5-flucytosine under the catalysis of deoxyribose transferase. The method avoids the operation of radical protection and deprotection during chemical synthesis, shortens synthesizing steps, improves the yield of target products, reduces synthesizing cost, and avoids environmental pollution brought by organic solvents used in chemical synthesizing methods.
Description
Technical field
The present invention relates to a kind of synthetic method of 5-flurocytosine nucleosides, relate to a kind of enzymatic synthesis method of 5-flurocytosine nucleosides specifically.
Background technology
β-D-2 ', 3 '-two deoxidations are two, and dehydrogenation-5-flurocytosine nucleosides (D-D4FC) is the s-generation does not have toxic side effect to the human body plastosome nucleoside analog (Schinazi, R.; Ma, L.; Shi, J.; Liotta, D.; Faraj, A.; Sommadossi, J.P.Abstract 557/41174,12th worldAIDS conf, Geneva, June 28-July 3,1998.), be a kind of HIV efficiently and HBV selective depressant, can suppress duplicating of HIV and HBV in vitro and in vivo.It is reported that it can be restrained AZT, ddC, ddI, D4T and 3TC have drug-fast HIV-1 C-type virus C, simultaneously, HBV are also had very strong supression effect (Hammond, J.; Schinazi, R.; Schlueter-Wirtz, S.; Mellors, J.Abstract 597,6th Conference on Retroviruses andOpportunistic Infections, Chicago, January 31-fuebuary4,1999.).Owing in cell system and body, show good pharmacy dynamics and hypotoxicity, be regarded as the potential good medicine of anti-HIV and HBV.This medicine is at the U.S., Germany and France's beginning phase ii clinical trial.
Up to the present, D-D4FC only has report (J Org Chem, 1992,57 (14): 3887 of chemical synthesis; JMed Chem, 1999,42 (5): 859; Bioorg Med Chem Lett, 1998,8 (22): 3245; J Med Chem, 1999,42 (5): 859; Carbohydrate Res.1997,300 (4): 375), the yield of these synthetic routes is all lower, and the mole total recovery is all about 10%; The condition of chemosynthesis is relatively harsher, needs operation at low temperatures, and the industrialization difficulty is bigger; In the process of chemosynthesis, need to use expendable heavy metal catalyst, not only toxic to workman's health, and also also very unfriendly to environment, do not meet the green friendly chemical theme of advocating now.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of synthetic method of 5-flurocytosine nucleosides, to solve synthetic cost height in the prior art, synthetic route is long, yield is low, photolytic activity is low shortcoming.
The invention discloses a kind of synthetic method of 5-flurocytosine nucleosides, this method comprises the steps:
The thymidine of structural formula in the reaction formula 1 is carried out the base permutoid reaction with 5-flurocytosine in the damping fluid that contains the ribodesose transferring enzyme, 37-42 ℃ was reacted 20-30 hour; The dried residue of reaction solution is the 5-flurocytosine nucleosides with methyl alcohol or dichloromethane extraction, the fluorine-containing nucleoside analog that obtains structural formula 2;
Wherein reaction formula is:
Wherein damping fluid is KH
2PO
4A kind of in damping fluid, citrate buffer solution or the water.
Add the ribodesose transferring enzyme in the reaction process.
Preferred synthetic method is:
The thymidine of 100-150mg50mmol is carried out the base permutoid reaction with the 5-flurocytosine of 100-150mg50mmol in the damping fluid that contains 4ml ribodesose transferring enzyme, 37-42 ℃ of constant temperature was with 100-200r/min tachyphylaxis 20-30 hour; The dried residue of reaction solution methyl alcohol or the dichloromethane extraction of 10-30ml, the fluorine-containing nucleoside analog that obtains structural formula 2 is the 5-flurocytosine nucleosides.
Adopt the synthetic D-D4FC of biological enzyme, have following advantage
(1) enzyme catalysis efficient height, the catalytic efficiency of enzyme exceeds 10 than general chemical catalysis
6～10
13Doubly.In enzymic catalytic reaction, avoided radical protection and de-protected operation in chemosynthesis, shortened the synthetic step, improved the yield of target product, simultaneously, reduced synthetic cost.Experimental result shows, has been brought up to about 25% by about 10% of chemical synthesis with the productive rate of the synthetic D-D4FC of enzyme process catalysis, and experimental procedure has shortened to a step by seven steps of chemical synthesis.
(2) good reaction selectivity, most of enzymes have the specificity of height, and a certain gene of catalysis that can be single-minded rapidly or the reaction of a certain specific position synthesize the asymmetric compound with high optical activity.Experimental result shows that the ee value of synthesizing D-D4FC with enzyme process catalysis has reached 99%
(3) carry out in the aqueous solution of temperature of reaction under normal temperature (37-42 ℃) normal pressure of the synthetic D-D4FC of enzyme catalysis, mild condition, control is easy, and side reaction is few, the environmental pollution of having avoided the organic solvent in the chemical synthesis to bring simultaneously.
Embodiment:
Embodiment 1
Get unsaturated thymidine (D-D4T) 1112mg (50mmol), 5-flurocytosine (5-FC) 124mg (100mmol), KH
2PO
4Damping fluid 30mL (pH6.35), the homemade ribodesose transferring enzyme of 4ml liquid send out with 150r/min vibration in water bath with thermostatic control and answer 20-24h in the Erlenmeyer flask of 100ml.Being reflected at boils in the boiling water boils 5min with stopped reaction.Reaction solution is frozen drying, and residue extracts with the 10ml ethanol/methylene, and extraction liquid launches with TLC, obtains product 214.96mg, Y:16.8%.
Embodiment 2
Get unsaturated thymidine (D-D4T) 1112mg (50mmol), 5-flurocytosine (5-FC) 124mg (100mmol), KH
2PO
4Damping fluid 30mL (pH6.35), the homemade ribodesose transferring enzyme of 4ml liquid send out with 150r/min vibration in water bath with thermostatic control and answer 20-24h in the Erlenmeyer flask of 100ml.Being reflected at boils in the boiling water boils 5min with stopped reaction.Reaction solution is frozen drying, residue 30ml methanol extraction, and concentrating under reduced pressure gets solid mixture.Solid mixture is crossed silica gel column chromatography (20%CH
3OH/CH
2Cl
2, v/v) purifying gets product 227.5mg, Y:24.6%.
Embodiment 3
Get unsaturated thymidine (D-D4T) 1112mg (50mmol), 5-flurocytosine (5-FC) 124mg (100mmol), citrate buffer solution 30mL (pH6.35), the homemade ribodesose transferring enzyme of 4ml liquid is sent out with 150r/min vibration in water bath with thermostatic control and is answered 20-24h in the Erlenmeyer flask of 100ml.Being reflected at boils in the boiling water boils 5min with stopped reaction.Reaction solution is frozen drying, residue 30ml methanol extraction, and concentrating under reduced pressure gets solid mixture.Solid mixture is crossed silica gel column chromatography (20%CH
3OH/CH
2Cl
2, v/v) purifying gets product 227.0mg, Y:24.1%.
The TLC method is adopted in qualitative analysis in the reaction process.With activated silica gel G plate is carrier, methylene chloride-methanol-water (8: 2: 0.25v/v) be developping agent, the ultraviolet colour developing.The R of D-D4T, D-D4FC, 5-FC and thymus pyrimidine
fReference value is respectively the analysis of 0.82,0.61,0.29 and 0.05 substrate and product and adopts the HPLC method.Adopt chromatographic column Diamonsil C
18, Φ 4.6 * 150mm; With the 15-35% methanol is that moving phase is carried out tonsure washing, flow velocity 0.9mL/min; Detect at the 270nm place.The t of D-D4FC, 5-FC, D-D4T and thymus pyrimidine
RReference value is respectively 7.06min, 5.20min, 3.86min and 2.71min.HPLC analyzes specimen in use and all takes from organic phase, and calculates productive rate according to substrate starting point concentration in the organic phase and production concentration.
The spectrum analysis data are as follows:
1H?NMR(DMSO-d
6)：δ8.04(d，J＝7.6Hz，1H，H-6)，7.77(bs，1H，NH)，7.64(bs，1H，NH)，6.82(bs，1H，H-1′)，6.39(d，J＝7.6Hz，1H，H-3′)，5.83(d，J＝6.0Hz，1h，H-2′)，5.11(t，J＝6.0Hz，1H，OH)，4.77(bs，1H，H-4′)，3.41-3.59(m，2H，H-5′)
19F?NMR(DMSO-d
6)：δ171.36(J＝8.0Hz)
EI?mass?calcd?for?C
9H
11FN
3O
3(M+H
+)：228.0784，found：228.0784
UV(MeOH)：max280nm
Claims (2)
1. the synthetic method of a 5-flurocytosine nucleosides is characterized in that this method comprises the steps:
The thymidine of structural formula in the reaction formula 1 is carried out the base permutoid reaction with 5-flurocytosine in the damping fluid that contains the ribodesose transferring enzyme, 37-42 ℃ was reacted 20-30 hour; The dried residue of reaction solution is the 5-flurocytosine nucleosides with methyl alcohol or dichloromethane extraction, the fluorine-containing nucleoside analog that obtains structural formula 2;
Wherein reaction formula is:
Wherein damping fluid is KH
2PO
4A kind of in damping fluid, citrate buffer solution or the water.
2. the synthetic method of 5-flurocytosine nucleosides according to claim 1 is characterized in that this method comprises the steps:
The thymidine of 100-150mg 50mmol is carried out the base permutoid reaction with the 5-flurocytosine of 100-150mg 50mmol in the damping fluid that contains 4ml ribodesose transferring enzyme, 37-42 ℃ of constant temperature was with 100-200r/min tachyphylaxis 20-30 hour; The dried residue of reaction solution obtains the 5-flurocytosine nucleosides with methyl alcohol or the dichloromethane extraction of 10-30ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200510029428XA CN100355901C (en) | 2005-09-06 | 2005-09-06 | Synthesis method of 5-fluorocytidine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200510029428XA CN100355901C (en) | 2005-09-06 | 2005-09-06 | Synthesis method of 5-fluorocytidine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1763213A CN1763213A (en) | 2006-04-26 |
| CN100355901C true CN100355901C (en) | 2007-12-19 |
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ID=36747532
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200510029428XA Expired - Fee Related CN100355901C (en) | 2005-09-06 | 2005-09-06 | Synthesis method of 5-fluorocytidine |
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|---|---|
| CN (1) | CN100355901C (en) |
-
2005
- 2005-09-06 CN CNB200510029428XA patent/CN100355901C/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| 酶法合成核苷类抗病毒药物 邱蔚然，丁庆豹.中国医药工业杂志，第30(10)期 1999 * |
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| CN1763213A (en) | 2006-04-26 |
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Granted publication date: 20071219 Termination date: 20100906 |