CN101307076A - Prodrug with liver-targeted anti-HBV effect - Google Patents
Prodrug with liver-targeted anti-HBV effect Download PDFInfo
- Publication number
- CN101307076A CN101307076A CNA2008101237339A CN200810123733A CN101307076A CN 101307076 A CN101307076 A CN 101307076A CN A2008101237339 A CNA2008101237339 A CN A2008101237339A CN 200810123733 A CN200810123733 A CN 200810123733A CN 101307076 A CN101307076 A CN 101307076A
- Authority
- CN
- China
- Prior art keywords
- adefovir
- ester
- tenofovir
- acid
- liver
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
The invention provides a prodrug of a hepatitis b virus resistant drug with a liver targeting action and a preparation method for the same. The prodrug is a prodrug of Adefovir or Tenofovir. Higher fatty acid, glyceride, cholic acid compounds or the base groups of derivatives thereof with the liver targeting action are connected with Adefovir or Tenofovir to prepare the prodrug. After being taken into a human body, the prodrug is concentrated in the liver and then is decomposed, thereby having the effects of reducing the dosage, improving the curative effect, reducing the drug concentration in a kidney and reducing the toxic and side effect. The invention also provides a method for synthesizing the prodrug.
Description
Technical field
What the present invention relates to is some hepatitis B virus resisting medicines with liver targeting and preparation method thereof, particularly prodrug of adefovir ester and tenofovir and preparation method thereof.
Background technology
Hepatitis B is the disease of serious harm human health, and the whole world has 2,000,000,000 people to infect hepatitis virus (HBV) approximately, hepatitis B virus carriers 3.5 hundred million people wherein, and China is hepatitis B country occurred frequently, the HBV carrier surpasses 1.2 hundred million, surplus the hepatitis B patient 3,000 ten thousand.Chronic viral hepatitis B patient liver cirrhosis incidence 2-20% develops into liver cancer and accounts for 6-15%.China chronic viral hepatitis B patient finally has 25-40% to die from liver cirrhosis or liver cancer.The HBV carrier finally dies from the danger of relevant hepatopathy, and the male sex is 50%, and the women is 15%.The medical fee that China is directly used in viral hepatitis every year exceeds hundred million yuan of 300-500.Therefore, the treating hepatitis B medicine becomes one of emphasis of global medical science research and development safely and effectively.
Hepatitis B resisting medicated mainly containing commonly used at present: alpha-interferon, lamivudine, adefovir ester, Entecavir and Telbivudine.Tenofovir disoproxil and emtricitabine go on the market as inverase in addition, and anti-hepatitis b is at III phase clinical stage.
More than several anti-hbv drug alpha-interferons be outside the immunostimulant, all the other all are ucleosides antiviral drugs.
Adefovir ester (I) and tenofovir disoproxil (II) are respectively the prodrugs of Adefovir (III) and tenofovir (IV), Adefovir and tenofovir structural similitude, be that the ucleosides antiviral drug has the anti-hepatitis b effect, but because ionization in vivo, enteric solubility difference and be difficult for to absorb, bioavailability is very low, can not directly use.Adefovir ester and tenofovir disoproxil then enteric solubility are good, and bioavailability is higher, become to be used for clinical medicine.
Structural formula is:
Nucleoside medicine occupies considerable status in hepatitis B virus resisting medicine, but curative effect is not really desirable, mainly be that nucleoside medicine need enter liver cell because hepatitis B core antigen is present in the liver cell, and through thymidine kinase catalysis triphosphoric acid rear produce effects.And nucleoside medicine be difficult for to see through liver cell, and the cell of hepatitis B virus infection lacks thymidine kinase, and the host cell phosphorylation is slow, does not reach the effective concentration that suppresses virus replication.Though heavy dose of administration can improve Plasma Concentration, increases curative effect, can be owing to occur toxic side effect such as blood urine and renal tubal dysfunction rapidly through renal excretion in blood by the medicine of phosphorylation.
1994, Hostetler KY etc. reported two oleoyl phosphatidyl zalcitabines (DOP-ddC) synthetic and liver target (Hostetler KY et al., Antiviral Res.1994,
24(1): 59-67), but the anti-HBV effect of ddC own is not strong, and deliberately justice is little.
Nineteen ninety-five, reports such as Xie H. have synthesized 1 of hepatitis B resisting medicated lamivudine, 2-dipalmitoyl-glycerol-3-phosphoric acid conjugates (Xie H.et al., Antiviral Res.1995,
28(2): 113-20), test shows that it has the liver target.2005, equality was reported the synthetic and preliminarily stabilised Journal of Sex Research (Hu equality, Central-South pharmacy, 2005 (1)) of lamivudine courage acid amides recklessly.But the present resistance of its parent medicine lamivudine is higher, has been worth little.
1996, Korba BA etc. reported 1, the liver target of 2-two palmityl phosphatidyl dideoxy guanosines (Korba BA et al., Hepatology, 1996,
23(5): 958-63), but the effect of parent medicine dideoxy guanosine anti-hepatitis b is also not really strong.
1997, Fiume L. etc. reported the conjugates (ara-AMP-L-poly-lys) of vidarabine acid and lactose amine-L-polylysine have liver target (Fiume L.et al., J.Hepatol.1997,
26(2): 253-9).The same year, Fiume L. etc. reported again the lactose amine human albumin conjugates of vidarabine have liver target (Fiume L.et al., Ital.J.Gastsoenterol Hepatol., 1997,
29(3): 275-80; Fiume L.et al., Viral Hepat.1997,
4(6): 363-70), but the problem that exists is that vidarabine itself is not effectively hepatitis B resisting medicated, and the structure of its prodrug is very complicated again.
1999, Hashida M. etc. reported PGE
1(PGE
1)-semi-lactosi hydrazides-polyglutamic acid conjugates has liver target and discharges PGE at hepatic parenchymal cells
1(Hashida M.et al., J.Control Release, 1999,
66(1-2): 253-62), problem also is that structure is too complicated.
2002, Xiang Guiming etc. reported the preparation of lactosylation polylysine conjugates (Lac-PLL-ACV) of acyclovir and liver target (Xiang Guiming etc., the The 2nd Army Medical College journal, 2002,
24(3): 311-3), shortcoming is that acyclovir itself is not good hepatitis B virus resisting medicine, and the conjugates structure is also complicated.
2004, Mark D.Erion etc. reported the design of the liver activation prodrug of vidarabine acid, lamivudine, cytosine arabinoside and Adefovir, synthetic and biological property (Mark D.Erion et al., J.Am.Chem.Soc., 2004,
126: 5154-63), be the six-ring phosphate compounds, wherein the prodrug Reomfovir of Adefovir has entered II phase clinical study, and this compounds is not to concentrate target to be distributed in the liver in vivo, and only is to activate in liver, and certain limitation is arranged, and is undesirable.
According to the above, the situation of target anti-hepatitis b prodrug research both at home and abroad can reduce:
1) target anti-hepatitis b prodrug research time of carrying out shorter, study lessly, system not only prepares and studies a few compounds very much, also has a lot of defectives.
2) to be that the parent medicine of prodrug is selected improper for one of defective, is not good hepatitis B resisting medicated as zalcitabine, dideoxy guanosine, vidarabine, acyclovir, PGE of clinical efficacy
1Deng, though that the parent medicine that has has been selected is clinical commonly used hepatitis B resisting medicated, as lamivudine, its Time To Market is longer, the resistance incidence is higher at present.
3) another defective is, the prodrug structure is too complicated, molecular weight is very big, be difficult for preparation and quality control, as be used for clinically, and dosage is also very big, and multiple molecule segment might produce other toxic side effect in vivo, lack practicality, as Adefovir-quadrol-Methionin-hydroxypentanoic acid-semi-lactosi conjugates.
4) also having a defective is or not that the liver target distributes in the body, but activation in the liver though the anti-hepatitis b effect is better, can not reduces dosage and reduce toxic side effect, as the six-ring phosphoric acid ester of vidarabine acid, lamivudine, cytosine arabinoside and Adefovir.
Summary of the invention
Technical problem: the present invention proposes weak point such as undesirable at the hepatitis B resisting medicated ubiquitous curative effect of ucleosides, that toxic side effect is bigger prodrug of anti-hepatitis B virus with liver targeting and preparation method thereof is provided, they can be used for treating hepatitis B, can improve curative effect, reduce consumption by its liver target effect, reduce toxic side effect.
Technical scheme:
The present invention proposes a kind of prodrug with liver-targeted anti-HBV effect, its structure is:
R wherein
1Be H or CH
3R
3Be H or R
2R
2Be R
4Or L-R
4, wherein L connects base, is C
1-C
8Alkyl or ether group, R
4It is the group of higher fatty acid with liver targeting, glyceryl ester, cholic acid compounds or derivatives thereof.
R
4Can also be preferably:
N=11-25; Or
L is preferably-CH
2-,-CH
2-CH
2-,-CH
2-CH
2-CH
2-,-CH
2-CH
2-CH
2-CH
2-or-CH
2-CH
2-O-CH
2-CH
2-.
Higher fatty acid can be preferably lauric acid, tetradecanoic acid, palmitinic acid, stearic acid, eicosanoic acid, docosoic or cerinic acid.
Glyceryl ester can be preferably 1-monopalmitin, stearin, 1, two palmitins of 2-or stearin.
The cholic acid class can be preferably cholic acid, lithocholic acid, Deoxycholic Acid, ursodesoxycholic acid or Chenodiol.
The preceding medicine of these that invented most preferably is:
Adefovir list palm acyl-oxygen methyl esters
The two palm acyl-oxygen methyl esters of Adefovir
Tenofovir list palm acyl-oxygen methyl esters
The two palm acyl-oxygen methyl esters of tenofovir
Adefovir list stearoyl keto methyl esters
Adefovir stearic bicine diester oxygen methyl esters
Tenofovir list stearoyl keto methyl esters
Tenofovir stearic bicine diester oxygen methyl esters
The bimonthly osmanthus of Adefovir acyl-oxygen methyl esters
The two hexacosane acyl-oxygen methyl esters of Adefovir
Adefovir list palmitoyl glyceride
The two palmitoyl glycerides of Adefovir
Tenofovir list palmitoyl glyceride
The two palmitoyl glycerides of tenofovir
Adefovir list stearyl glyceryl ester
Adefovir stearic bicine diester glyceryl ester
Tenofovir list stearyl glyceryl ester
Tenofovir stearic bicine diester glyceryl ester
Adefovir Methyl cholate-3-ester
Adefovir Ethyl cholate-3-ester
Tenofovir Methyl cholate-3-ester
Tenofovir Ethyl cholate-3-ester
Adefovir courage acyl glycol ester
Adefovir courage acyl propylene glycol ester
Adefovir courage acyl butanediol ester
Adefovir courage acyl binaryglycol ester
Tenofovir courage acyl glycol ester
Tenofovir courage acyl binaryglycol ester
Adefovir list ursodeoxycholic acid propylene glycol ester
Tenofovir list ursodeoxycholic acid propylene glycol ester
Adefovir list Chenodiol butanediol ester
Adefovir list lithocholic acid glycol ether ester
The invention allows for the preparation method of these prodrugs:
A kind of preparation has the preceding medicament preparation with liver-targeted anti-HBV effect of higher fatty acid compound or derivatives thereof group, comprise following steps: higher fatty acid is made the chloromethyl ester derivative earlier, again in N-Methyl pyrrolidone or DMF (dimethyl formamide) under the triethylamine effect with adefovir ester or tenofovir condensation.
The synthetic route of the senior fatty acyl-oxygen methyl esters of Adefovir or tenofovir is:
R=C
11-C
25Alkyl
R
1＝H，CH
3
R
2＝H，-CH
2OOCR
Temperature of reaction is 50-120 ℃, and the best is 70-90 ℃.
A kind of preparation method of the prodrug with liver-targeted anti-HBV effect of the group that the glyceride compounds or derivatives thereof arranged comprises following steps: glyceryl ester in N-Methyl pyrrolidone or DMF (dimethyl formamide) in DCC (dicyclohexylcarbodiimide) effect down and adefovir ester and tenofovir condensation.
The synthetic route of the senior fatty acyl glyceryl ester of Adefovir or tenofovir is:
R
1＝H，CH
3
R
2=H, C
12-C
26Senior fatty acyl group
R
3=C
12-C
26Senior fatty acyl group
Temperature of reaction is 50-150 ℃, and the best is 80-120 ℃.
A kind of preparation method of the prodrug with liver-targeted anti-HBV effect of the group that cholic acid compounds or derivatives thereof arranged, comprise following steps: the carboxyl on the cholic acid and methyl alcohol generate Methyl cholate, and hydroxyl on its 3 and the phosphonic acids on Adefovir or the tenofovir are condensed into ester; Or the carboxyl on the cholic acid earlier with ethylene glycol, 1, ammediol, 1, the reaction of glycol such as 4-butyleneglycol, glycol ether generates ester, another hydroxyl in the glycol again with Adefovir or tenofovir on phosphonic acids be condensed into ester.
Adefovir or tenofovir become the synthetic route of ester to be with Methyl cholate or ethyl ester:
R
1＝H，CH
3
R
2, R
3=H or CH
3
R
4＝CH
3、C
2H
5
Temperature of reaction is 50-150 ℃, and the best is 80-120 ℃.
Adefovir or tenofovir become the synthetic route of ester to be with cholic acid class diol ester:
R
1＝H，CH
3
R
2, R
3=H or CH
3
R
4＝CH
2CH
2、CH
2CH
2CH
2、CH
2CH
2CH
2、CH
2CH
2OCH
2CH
2
Temperature of reaction is 20-150 ℃, and the best is 50-120 ℃.
Beneficial effect: treat the curative effect of hepatitis B and reduce its toxicity in order to improve nucleoside medicine, the hepatitis B resisting medicated imagination of target is proposed, on hepatitis B medicine, connect molecule and make prodrug with liver avidity, after taking in the body it is enriched in the liver and decomposes again, can play simultaneously like this and reduce consumption, raising curative effect, the effect that the kidney concentration reduces toxic side effect that reduces.Experimental results show that but this compound of invention is synthetic and has the hepatitis B resisting medicated effect of target.The problems of the prior art have been solved.
The target of target anti-hepatitis b prodrug of the present invention and carrier are higher fatty acid class, glyceride type and cholic acid class.One of these carriers and the coupling of one of parent medicine use no or little other and connect base, so that the prodrugs structure is not too complicated, are easy to preparation and quality control, are easy to use.
Target anti-hepatitis b prodrug of the present invention because selectivity is concentrated and to be distributed in the liver, can improve curative effect, reduce the toxicity of other organ, especially kidney, also can suitably reduce dosage.
Target anti-hepatitis b prodrug parent medicine of the present invention is connected with the form of targeting vector with ester, helps improving stability to hydrolysis and the liver target of prodrug in gi tract.
Target anti-hepatitis b prodrug of the present invention is fat-soluble, and oral easy absorption has high bioavailability.Target anti-hepatitis b prodrug of the present invention can decomposite the parent medicine with certain speed in liver, produce anti-HBV effect, has guaranteed drug effect.
Embodiment
Embodiment 1: the preparation and the evaluation of the two palm acyl-oxygen methyl esters of Adefovir
250mL there-necked flask dress agitator, prolong, thermometer add palmitinic acid 52g, are warmed up to 80 ℃ of solid dissolvings, add several DMF, stir down and slowly splash into sulfur oxychloride 50mL, have hydrogen chloride gas to emit.Finish restir 2 hours, and got yellow clear liquor.Add Paraformaldehyde 96 6g and Zinc Chloride Anhydrous 0.6g, stirred 3 hours, Paraformaldehyde 96 dissolves gradually.Reaction solution is poured in the frozen water, adds sodium hydroxide solution under stirring and transfers to neutrality, oily matter ethyl acetate extraction, the washing of ester layer, anhydrous sodium sulfate drying.Evaporated under reduced pressure, solidify palmitinic acid chloromethyl ester off-white color solid 58g.
Add Adefovir 1.0g, palmitinic acid chloromethyl ester 3.6g, N-Methyl pyrrolidone 20mL and triethylamine 1mL in the 100mL round-bottomed flask, 75 ℃ of following stirring reactions 4 hours.Add the 50mL cold ethyl acetate, stir elimination white solid after 1 hour, the filtrate water washing is made sand behind the ester phase anhydrous sodium sulfate drying, is that eluent carries out column chromatography for separation with ethyl acetate/dehydrated alcohol=3/1, gets off-white color solid 0.5g.MS-ESI(+)：810[M+H]
+。
Embodiment 2: the preparation and the evaluation of the two palm acyl-oxygen methyl esters of tenofovir
The same embodiment 1 method replaces the Adefovir reaction with tenofovir, gets the off-white color solid.
MS-ESI(+)：824[M+H]
+。
Embodiment 3: the preparation and the evaluation of Adefovir list palm acyl-oxygen methyl esters
Add Adefovir 1.0g, palmitinic acid chloromethyl ester 1.1g, N-Methyl pyrrolidone 20mL and DCC1g in the 100mL round-bottomed flask, 50 ℃ of following stirring reactions 3 hours.Add the 50mL ethyl acetate, use the 1M sodium hydroxide solution extraction, extracting solution with in the hydrochloric acid with after use ethyl acetate extraction again, make sand behind the ester phase anhydrous sodium sulfate drying, be that eluent carries out column chromatography for separation with ethyl acetate/dehydrated alcohol, must off-white color solid 0.4g.MS-ESI(-)：540[M-H]
-。
Embodiment 4: the preparation and the evaluation of tenofovir list palm acyl-oxygen methyl esters
The same embodiment 3 methods replace the Adefovir reaction with tenofovir, get off-white color solid 0.3g.MS-ESI(-)：554[M-H]
-。
Embodiment 5: the preparation and the evaluation of Adefovir stearic bicine diester oxygen methyl esters
250mL there-necked flask dress agitator, prolong, thermometer add stearic acid 58g, are warmed up to 80 ℃ of solid dissolvings, add several DMF, stir down and slowly splash into sulfur oxychloride 50mL, have hydrogen chloride gas to emit.Finish restir 3 hours, and got yellow liquid.Add Paraformaldehyde 96 6g and Zinc Chloride Anhydrous 0.6g, stirred 3 hours, Paraformaldehyde 96 dissolves gradually.Reaction solution is poured in the frozen water, stirs down and transfers neutrality with sodium hydroxide solution, oily matter ethyl acetate extraction, the washing of ester layer, anhydrous sodium sulfate drying.Evaporated under reduced pressure, solidify stearic acid chloromethyl ester off-white color solid 61g.
Add Adefovir 1.1g, stearic acid chloromethyl ester 3.2g, DMF20mL and triethylamine 1mL in the 100mL round-bottomed flask, 80 ℃ of following stirring reactions 4 hours.Add the 50mL cold ethyl acetate, stir elimination solid after half an hour, the filtrate water washing is made sand behind the ester phase anhydrous sodium sulfate drying, is that eluent carries out column chromatography for separation with ethyl acetate/dehydrated alcohol=3/1, off-white color solid 0.8g.MS-ESI(+)：866[M+H]
+。
Embodiment 6: the preparation and the evaluation of tenofovir stearic bicine diester oxygen methyl esters
The same embodiment 5 methods replace the Adefovir reaction with tenofovir, and 90 ℃ of temperature of reaction get off-white color solid 0.7g.MS-ESI(+)：880[M+H]
+。
Embodiment 7: the preparation and the evaluation of Adefovir list stearoyl keto methyl esters
With example 3 methods, replace the reaction of palmitinic acid chloromethyl ester with the stearic acid chloromethyl ester, get off-white color solid 0.5g, MS-ESI (-): 568[M-H]
-
Embodiment 8: the preparation and the evaluation of tenofovir list stearoyl keto methyl esters
The same embodiment 7 methods replace the Adefovir reaction with tenofovir, get off-white color solid 0.5g, MS-ESI (-): 582[M-H]
-
Embodiment 9: the preparation and the evaluation of the bimonthly osmanthus of Adefovir acyl-oxygen methyl esters
The same embodiment 1 method replaces the reaction of palmitinic acid chloromethyl ester with the lauric acid chloromethyl ester, gets the off-white color solid.MS-ESI(+)：698[M+H]
+。
Embodiment 10: the preparation and the evaluation of the two hexacosane acyl-oxygen methyl esters of Adefovir
The same embodiment 1 method replaces the reaction of palmitinic acid chloromethyl ester with the hexacosanoic acid chloromethyl ester, and 120 ℃ of temperature of reaction get the off-white color solid.MS-ESI(+)：1091[M+H]
+。
Embodiment 11: the preparation and the evaluation of Adefovir list palmitoyl glyceride
Add monopalmitin 7g (0.02mol), Adefovir 2.73g (0.01mol), N-Methyl pyrrolidone 30mL and N in the 100mL round-bottomed flask, N-dicyclohexyl carbimide 4.1g (0.02mol), reacted 6 hours down at 100 ℃, the pressure reducing and steaming solvent, add the 50mL dehydrated alcohol, with the insoluble urea elimination that generates, filter cake is washed with an amount of dehydrated alcohol, merge ethanol liquid, add 10g silica gel system sand.Cross post with methanol/ethanol=1/3 for elutriant, evaporated under reduced pressure gets off-white color solid 2.6g, MS-ESI (-): 584[M-H]
-
Embodiment 12: the preparation and the evaluation of tenofovir list palmitoyl glyceride
The same embodiment 11 methods replace the Adefovir reaction with tenofovir, get off-white color solid 1.9g, MS-ESI (-): 598[M-H]
-
Embodiment 13: the preparation and the evaluation of Adefovir list stearyl glyceryl ester
The same embodiment 11 methods replace the monopalmitin reaction with glyceryl monostearate, and 150 ℃ of temperature of reaction get off-white color solid 2.9g, MS-ESI (-): 612[M-H]
-
The preparation and the evaluation of embodiment 14 tenofovir list stearyl glyceryl ester
With embodiment 13 methods, replace the Adefovir reaction with tenofovir, get off-white color solid 2.9g, MS-ESI (-): 626[M-H]
-
Embodiment 15: the preparation and the evaluation of the two palmitoyl glycerides of Adefovir
Add glycerol-1,3-dipalmitate 11.4g (0.02mol), Adefovir 2.73g (0.01mol), DMF30mL and triethylamine 1.4mL in the 100mL round-bottomed flask, reacted 5 hours down at 120 ℃, the pressure reducing and steaming solvent, add the 50mL dehydrated alcohol, with the insoluble urea elimination that generates, filter cake is washed with an amount of dehydrated alcohol, merges ethanol liquid, adds proper silica gel system sand.Cross post with methanol/ethanol=1/3 for elutriant, evaporated under reduced pressure gets off-white color solid 3.6g, MS-ESI (-): 822[M-H]
-
Embodiment 16: the preparation and the evaluation of the two palmitoyl glycerides of tenofovir
The same embodiment 15 methods replace the Adefovir reaction with tenofovir, get off-white color solid 2.9g, MS-ESI (-): 836[M-H]
-
Embodiment 17: the preparation and the evaluation of Adefovir stearic bicine diester glyceryl ester
The same embodiment 15 methods replace the reaction of glycerol-1,3-dipalmitate Wei with distearin, and 150 ℃ of temperature of reaction get off-white color solid 2.5g, MS-ESI (-): 878[M-H]
-
Embodiment 18: the preparation and the evaluation of tenofovir stearic bicine diester glyceryl ester
The same embodiment 18 methods replace the Adefovir reaction with tenofovir, get off-white color solid 2.5g, MS-ESI (-): 892[M-H]
-
Embodiment 19: the preparation and the evaluation of Adefovir list courage acyl methyl esters
Add 60mL methyl alcohol and 10g cholic acid in the 100mL round-bottomed flask, splash into several concentrated hydrochloric acids again, refluxed 6 hours.Crystal is slowly separated out in cooling, and filtration drying gets white crystals 8.5g.
Add Methyl cholate 5g, Adefovir 2.2g, N-Methyl pyrrolidone 30mL, triethylamine 2mL and N in the 100mL round-bottomed flask, N-dicyclohexyl carbimide 2.2g, 80 ℃ of stirring reactions 4 hours.Oil pump pressure reducing and steaming solvent adds the 40mL dehydrated alcohol and refluxes half an hour insolubles elimination, ethanol liquid system sand.Column chromatography with methanol/ethanol mixed solvent gradient elution, gets off-white color solid 3.2g.MS-ESI(-)：676[M-H]
-。
Embodiment 20: the preparation and the evaluation of tenofovir list courage acyl methyl esters
The same embodiment 19 methods replace the Adefovir reaction with tenofovir, get the off-white color solid.MS-ESI(-)：690[M-H]
-。
Embodiment 21: the preparation and the evaluation of Adefovir list courage acyl ethyl ester
Add 100mL ethanol and 10g cholic acid in the 100mL round-bottomed flask, splash into several concentrated hydrochloric acids again, refluxed 6 hours.Crystal is slowly separated out in cooling, and filtration drying gets white crystals 8.5g.
Add Ethyl cholate 5g, Adefovir 2.2g, N-Methyl pyrrolidone 30mL, triethylamine 2mL and N in the 100mL round-bottomed flask, N-dicyclohexyl carbimide 2.2g, 100 ℃ of stirring reactions 4 hours.Oil pump pressure reducing and steaming solvent adds the 40mL dehydrated alcohol and refluxes half an hour insolubles elimination, ethanol liquid system sand.Column chromatography with methanol/ethanol mixed solvent gradient elution, gets off-white color solid 3.4g.MS-ESI(-)：690[M-H]
-。
Embodiment 22: the preparation and the evaluation of tenofovir list courage acyl ethyl ester
The same embodiment 21 methods replace the Adefovir reaction with tenofovir, get the off-white color solid.MS-ESI(-)：704[M-H]
-。
Embodiment 23: the preparation and the evaluation of Adefovir list courage acyl glycol ester
Add 50mL ethylene glycol and 10g cholic acid in the 100mL round-bottomed flask, splash into the 2mL concentrated hydrochloric acid again, 60 ℃ of stirring reactions 6 hours.Pour in the 300mL frozen water after the cooling slightly, the oily matter ethyl acetate extraction is washed 3 times again, and anhydrous sodium sulfate drying boils off solvent and gets light yellow oil 10.3g.Be directly used in next step.
Add cholic acid glycol ester 6g, Adefovir 2.0g, triethylamine 2ml and N-Methyl pyrrolidone 20mL in the 100mL round-bottomed flask, stirring reaction is 3 hours in 80 ℃ of oil baths.Add DCC2.5g again and continue reaction 20 hours, oil pump pressure reducing and steaming solvent adds the 40mL dehydrated alcohol and refluxed 1 hour.The insolubles elimination, filtrate system sand, column chromatography, the methanol/ethanol gradient elution gets off-white color solid 2.1g.MS-ESI(-)：706[M-H]
-。
Embodiment 24: the preparation and the evaluation of tenofovir list courage acyl glycol ester
The same embodiment 23 methods replace the Adefovir reaction with tenofovir, get the off-white color solid.MS-ESI(-)：720[M-H]
-。
Embodiment 25: the preparation and the evaluation of Adefovir list ursodeoxycholic acyl propylene glycol ester
With the method for preparing the cholic acid glycol ester among the embodiment 23, with ursodesoxycholic acid and propylene glycol reaction system row ursodeoxycholic acid propylene glycol ester.Make the light brown solid 1.1g of Adefovir list ursodeoxycholic acid propylene glycol ester again, MS-ESI (-): 704[M-H]
-
Embodiment 26: the preparation and the evaluation of tenofovir list ursodeoxycholic acyl propylene glycol ester
The same embodiment 25 replaces the Adefovir reaction with tenofovir, and 100 ℃ of temperature of reaction get pale solid.MS-ESI(-)：718[M-H]
-。
Embodiment 27: the preparation and the evaluation of Adefovir list courage acyl butanediol ester
Add 60mL1 in the 100mL round-bottomed flask, 4-butyleneglycol and 10g cholic acid splash into the 2mL concentrated hydrochloric acid again, 70 ℃ of stirring reactions 6 hours.Pour in the 300mL frozen water after the cooling slightly, the oily matter ethyl acetate extraction is washed 3 times again, and anhydrous sodium sulfate drying boils off solvent and gets light yellow oil 12.6g.Be directly used in next step.
Add cholic acid butanediol ester 8g, Adefovir 1.5g, triethylamine 3ml and N-Methyl pyrrolidone 30mL in the 100mL round-bottomed flask, stirring reaction is 3 hours in 80 ℃ of oil baths.Add DCC2g again and continue reaction 20 hours, oil pump pressure reducing and steaming solvent adds the 30mL dehydrated alcohol and refluxed 1 hour.The insolubles elimination, filtrate system sand, column chromatography, the methanol/ethanol gradient elution gets off-white color solid 1.6g.MS-ESI(-)：734[M-H]
-。
Embodiment 28: the preparation and the evaluation of tenofovir list cholic acid butanediol ester
The same embodiment 27 methods replace the Adefovir reaction with tenofovir, get the off-white color solid.MS-ESI(-)：748[M-H]
-。
Embodiment 29: the preparation and the evaluation of Adefovir list goose deoxidation courage acyl butanediol ester
With embodiment 27 methods, substitute the cholic acid reaction with Chenodiol, get the off-white color solid, MS-ESI (-): 718[M-H]
-
Embodiment 30: the preparation and the evaluation of Adefovir list courage acyl glycol ether ester
Add 50mL glycol ether and 10g cholic acid in the 100mL round-bottomed flask, splash into the 1mL concentrated hydrochloric acid again, 80 ℃ of stirring reactions 5 hours.Pour in the 300mL frozen water after the cooling slightly, the oily matter ethyl acetate extraction is washed 3 times again, and anhydrous sodium sulfate drying boils off solvent and gets light yellow oil 10.5g.Be directly used in next step.
Add cholic acid glycol ether ester 5g, Adefovir 1.5g, triethylamine 2ml and N-Methyl pyrrolidone 20mL in the 100mL round-bottomed flask, stirring reaction is 5 hours in 100 ℃ of oil baths.Add DCC2g again and continue reaction 20 hours, oil pump pressure reducing and steaming solvent adds the 30mL dehydrated alcohol and refluxed 1 hour.The insolubles elimination, filtrate system sand, column chromatography, the methanol/ethanol gradient elution gets off-white color solid 1.8g.MS-ESI(-)：750[M-H]
-。
Embodiment 31: the preparation and the evaluation of tenofovir list courage acyl glycol ether ester
The same embodiment 30 methods replace the Adefovir reaction with tenofovir, get the off-white color solid.MS-ESI(-)：764[M-H]
-。
Embodiment 32: the preparation and the evaluation of Adefovir list stone courage acyl glycol ether ester
The same embodiment 30 methods replace the cholic acid reaction with lithocholic acid, and 150 ℃ of temperature of reaction get the off-white color solid.MS-ESI(-)：718[M-H]
-。
Embodiment 33: the activity test of prodrug representation compound
Get the rat that body weight is 160g-200g, raise two days later and use, by 2.0 * 10
-5Mol/kg body weight gastric infusion, after the administration respectively at sacrificed by exsanguination behind 0.5h, the 1.0h (♂ half and half for six animals of every time point, ♀).Liver, kidney are taken out in blood sampling rapidly, blot surperficial crimson blood, get tissue sample (about 0.5-1g), and precision weighing was in 4: 1 (ml: g) make homogenate with the high speed dispersion device behind the ratio adding physiological saline.Every duplicate samples is got 0.5ml homogenate before measuring, and adds internal standard substance acyclovir solution 20 ū l (being equivalent to acyclovir 2.0 ū g), mixing, add 10% perchloric acid 0.2ml, vibration 1min, the centrifugal 10min of 4000rpm, get supernatant 10 ū l sample introductions, get blank plasma simultaneously and tissue compares.The content of Adefovir in the assay determination computation organization (Tai Luofuwei).
Dosage=rat body weight * 2.0 * 10 wherein
-5Mol/kg * medicine molecular weight
The result is as follows:
From last table, can clearly find out, mouse stomach give among the present invention mentioned before behind the medicine, detect antiviral ingredients content in the liver obviously than the height of control drug, illustrate that prodrugs of the present invention has tangible liver targeting; Simultaneously as can be seen, the content in the kidney is obviously low than contrast content, proves that drug molecule has the effect of the renal toxicity of reduction.
Claims (10)
1, a kind of prodrug with liver-targeted anti-HBV effect, its structure is:
R wherein
1Be H or CH
3R
3Be H or R
2R
2Be R
4Or L-R
4, wherein L connects base, is C
1-C
8Alkyl or ether group, R
4It is the group of higher fatty acid with liver targeting, glyceryl ester, cholic acid compounds or derivatives thereof.
2, according to the described a kind of liver-targeted anti-HBV effect prodrug that has of claim 1, be characterised in that R
4For:
N=11-25; Or
N=11-25; Or
The group of cholic acid compounds or derivatives thereof.
3, according to the described a kind of liver-targeted anti-HBV effect prodrug that has of claim 1, be characterised in that-L-is :-CH
2-,-CH
2-CH
2-,-CH
2-CH
2-CH
2-,-CH
2-CH
2-CH
2-CH
2-or-CH
2-CH
2-O-CH
2-CH
2-.
4, according to claim 1,2 or 3 described a kind of liver-targeted anti-HBV effect prodrugs that have, be characterised in that higher fatty acid is lauric acid, tetradecanoic acid, palmitinic acid, stearic acid, eicosanoic acid, docosoic or cerinic acid.
5,, be characterised in that glyceryl ester is the single palmitoyl glyceride of 1-, stearin, 1, two palmitoyl glycerides of 2-or stearin according to claim 1,2 or 3 described a kind of prodrugs with liver-targeted anti-HBV effect.
6, according to claim 1,2 or 3 described a kind of prodrugs, be characterised in that the cholic acid class is cholic acid, lithocholic acid, Deoxycholic Acid, ursodesoxycholic acid or Chenodiol with liver-targeted anti-HBV effect.
7, according to claim 1,2 or 3 described a kind of prodrugs, be characterised in that these prodrugs are with liver-targeted anti-HBV effect:
Adefovir list palm acyl-oxygen methyl esters
The two palm acyl-oxygen methyl esters of Adefovir
Tenofovir list palm acyl-oxygen methyl esters
The two palm acyl-oxygen methyl esters of tenofovir
Adefovir list stearoyl keto methyl esters
Adefovir stearic bicine diester oxygen methyl esters
Tenofovir list stearoyl keto methyl esters
Tenofovir stearic bicine diester oxygen methyl esters
The bimonthly osmanthus of Adefovir acyl-oxygen methyl esters
The two hexacosane acyl-oxygen methyl esters of Adefovir
Adefovir list palmitoyl glyceride
The two palmitoyl glycerides of Adefovir
Tenofovir list palmitoyl glyceride
The two palmitoyl glycerides of tenofovir
Adefovir list stearyl glyceryl ester
Adefovir stearic bicine diester glyceryl ester
Tenofovir list stearyl glyceryl ester
Tenofovir stearic bicine diester glyceryl ester
Adefovir Methyl cholate-3-ester
Adefovir Ethyl cholate-3-ester
Tenofovir Methyl cholate-3-ester
Tenofovir Ethyl cholate-3-ester
Adefovir courage acyl glycol ester
Adefovir courage acyl propylene glycol ester
Adefovir courage acyl butanediol ester
Adefovir courage acyl binaryglycol ester
Tenofovir courage acyl glycol ester
Tenofovir courage acyl binaryglycol ester
Adefovir list ursodeoxycholic acid propylene glycol ester
Tenofovir list ursodeoxycholic acid propylene glycol ester
Adefovir list Chenodiol butanediol ester
Adefovir list lithocholic acid glycol ether ester.
8, a kind of a kind of preparation method as claimed in claim 4 with prodrug of liver-targeted anti-HBV effect, comprise following steps: higher fatty acid is made the chloromethyl ester derivative, again in N-Methyl pyrrolidone or dimethyl formamide under the triethylamine effect with adefovir ester or tenofovir condensation.
9, a kind of a kind of preparation method with prodrug of liver-targeted anti-HBV effect as claimed in claim 5 comprises following steps: glyceryl ester in N-Methyl pyrrolidone or dimethyl formamide under the dicyclohexylcarbodiimide effect with adefovir ester or tenofovir condensation.
10, a kind of a kind of preparation method as claimed in claim 6 with prodrug of liver-targeted anti-HBV effect, comprise following steps: the carboxyl on the cholic acid and methyl alcohol generate Methyl cholate, and hydroxyl on its 3 and the phosphonic acids on Adefovir or the tenofovir are condensed into ester; Or the carboxyl on the cholic acid earlier with ethylene glycol, 1, ammediol, 1, the reaction of glycol such as 4-butyleneglycol, glycol ether generates ester, another hydroxyl in the glycol again with Adefovir or tenofovir on phosphonic acids be condensed into ester.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101237339A CN101307076A (en) | 2008-05-30 | 2008-05-30 | Prodrug with liver-targeted anti-HBV effect |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101237339A CN101307076A (en) | 2008-05-30 | 2008-05-30 | Prodrug with liver-targeted anti-HBV effect |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101307076A true CN101307076A (en) | 2008-11-19 |
Family
ID=40123777
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008101237339A Pending CN101307076A (en) | 2008-05-30 | 2008-05-30 | Prodrug with liver-targeted anti-HBV effect |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101307076A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2017530194A (en) * | 2014-09-28 | 2017-10-12 | フアフイ ヘルス リミテッドＨｕａｈｕｉ Ｈｅａｌｔｈ Ｌｔｄ． | Inhibition of hepatitis B and D viruses and NTCP transport by polymerized bile acid derivatives |
| US9908908B2 (en) | 2012-08-30 | 2018-03-06 | Jiangsu Hansoh Pharmaceutical Co., Ltd. | Tenofovir prodrug and pharmaceutical uses thereof |
| JP2019069984A (en) * | 2013-03-15 | 2019-05-09 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Acrylic nucleoside phosphonic diester |
| WO2022168884A1 (en) * | 2021-02-04 | 2022-08-11 | 塩野義製薬株式会社 | Cationic lipid |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1947796A (en) * | 2005-10-13 | 2007-04-18 | 江苏正大天晴药业股份有限公司 | Chemical modified adefovir and tynofovir |
| CN101104625A (en) * | 2006-07-11 | 2008-01-16 | 北京世桥生物制药有限公司 | Liver targetable adefovir precursor medicament and medical use thereof |
| CN101182331A (en) * | 2007-11-30 | 2008-05-21 | 中国药科大学 | Adefovir dipivoxil ester colalin derivatives as well as preparation method and uses thereof |
-
2008
- 2008-05-30 CN CNA2008101237339A patent/CN101307076A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1947796A (en) * | 2005-10-13 | 2007-04-18 | 江苏正大天晴药业股份有限公司 | Chemical modified adefovir and tynofovir |
| CN101104625A (en) * | 2006-07-11 | 2008-01-16 | 北京世桥生物制药有限公司 | Liver targetable adefovir precursor medicament and medical use thereof |
| CN101182331A (en) * | 2007-11-30 | 2008-05-21 | 中国药科大学 | Adefovir dipivoxil ester colalin derivatives as well as preparation method and uses thereof |
Non-Patent Citations (3)
| Title |
|---|
| SAMIRA BENZARIA,ET AL.: ""Synthesis, in Vitro Antiviral Evaluation, and Stability Studies of Bis(S-acyl-2-thioethyl) Ester Derivatives of 9-[2-(Phosphonomethoxy)ethyl]adenine (PMEA) as Potential PMEA Prodrugs with Improved Oral Bioavailability"", 《JOURNAL OF MEDICINAL CHEMISTRY》 * |
| SCOTT J. HECKER ET AL.: ""Prodrugs of Phosphates and Phosphonates"", 《JOURNAL OF MEDICINAL CHEMISTRY》 * |
| XIAO ZHONG FU， ET AL.: ""Design, synthesis and in vitro evaluation of L-amino acid esters prodrugs of acyclic nucleoside phosphonates as anti-HBV agent"", 《CHINESE CHEMICAL LETTERS》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9908908B2 (en) | 2012-08-30 | 2018-03-06 | Jiangsu Hansoh Pharmaceutical Co., Ltd. | Tenofovir prodrug and pharmaceutical uses thereof |
| JP2019069984A (en) * | 2013-03-15 | 2019-05-09 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Acrylic nucleoside phosphonic diester |
| JP2017530194A (en) * | 2014-09-28 | 2017-10-12 | フアフイ ヘルス リミテッドＨｕａｈｕｉ Ｈｅａｌｔｈ Ｌｔｄ． | Inhibition of hepatitis B and D viruses and NTCP transport by polymerized bile acid derivatives |
| JP2019077710A (en) * | 2014-09-28 | 2019-05-23 | フアフイ ヘルス リミテッドＨｕａｈｕｉ Ｈｅａｌｔｈ Ｌｔｄ． | Polymeric bile acid derivatives that inhibit hepatitis b and d viruses and ntcp transport |
| US11701369B2 (en) | 2014-09-28 | 2023-07-18 | Huahui Health Ltd. | Polymeric bile acid derivatives inhibit Hepatitis B and D virus and NTCP transport |
| WO2022168884A1 (en) * | 2021-02-04 | 2022-08-11 | 塩野義製薬株式会社 | Cationic lipid |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102675403B (en) | Synthesis of anti-hepatitis B medicine LQC-X and application thereof | |
| CN103113264B (en) | Magnolol derivative, honokiol derivative and preparation method and application thereof | |
| CN104672290B (en) | A kind of medicine of disease for preventing or treating FXR mediations and its production and use | |
| CN105985396A (en) | Deuterated chenodeoxycholic acid derivative and pharmaceutical composition containing same | |
| CN105457038A (en) | Quick release type medicine phosphatide compound and medicine composition thereof | |
| CN106380502A (en) | G-protein-coupled receptor 5 (TGR5) modulators and methods of use thereof | |
| CN106866572B (en) | Nitric oxide donator type β elemene derivatives and its production and use | |
| CN101307076A (en) | Prodrug with liver-targeted anti-HBV effect | |
| CN104292290A (en) | Bile acid-drug conjugate with amino acid as connexon, and medical application thereof | |
| CN104707148A (en) | Polyethylene glycol modified glycyrrhetinic acid and curcumin compound used for resisting hepatic carcinoma, and preparation method thereof | |
| CN103864791B (en) | A kind of Entecavir derivative and preparation method thereof | |
| CN105131277A (en) | Polymer material containing cholic acid and liposome modified by same | |
| CN100513415C (en) | Bile acid derivative and pharmaceutical use thereof | |
| CN1911450A (en) | Live target adjuvant containing D-galactose and sterol or aliphatic alcohol and its preparation | |
| CN101580530A (en) | Amino acid conjugate prodrug of pentacyclic triterpenoid and medical application thereof | |
| CN104610240A (en) | Bicyclol-carnosine conjugate, and preparation method and application thereof | |
| CN101134769A (en) | Ursodeoxycholic acid entecavir acidamide and preparation method and use thereof | |
| CN1947796B (en) | Chemical modified adefovir and tynofovir | |
| CN106631957A (en) | Antitumor compound targeting FAP-alpha enzyme and preparation method and application thereof | |
| CN102617679B (en) | Preparation method and application of connected conjugated linoleic acid and gemcitabine prodrug | |
| US10729713B2 (en) | Pharmaceutical composition for treating hepatitis, liver fibrosis, and liver cancer | |
| CN100467480C (en) | Nitrate derivative of bile acid and its medical use | |
| WO2004026298A1 (en) | Derivatives of triptolide having high immunosuppressive effect and high water solubility, and uses thereof | |
| CN101774921A (en) | Method for preparing dicaffeoylquinic acid methyl compound and composition thereof | |
| CN107735108A (en) | A kind of PEGylation vitamin E periplocymarin conjugate nanoparticle and its preparation method and purposes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: WUZHONG MEDICINE DEVELOPMENT CO., LTD., JIANGSU Free format text: FORMER OWNER: QIN YINLIN Effective date: 20110701 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 210016 NO. 323-2 (1/F, BUILDING 1, NO. 12, FRONT BANSHANYUAN), ZHONGSHAN EAST ROAD, NANJING CITY, JIANGSU PROVINCE TO: 210009 NO. 26, MAJIA STREET, NANJING CITY, JIANGSU PROVINCE |
|
| TA01 | Transfer of patent application right |
Effective date of registration: 20110701 Address after: 210009 No. 26, Ma Jia street, Jiangsu, Nanjing Applicant after: Jiangsu Wuzhong Pharmaceutical Development Co., Ltd. Address before: 210016 Jiangsu Province, Nanjing city Zhongshan Road No. 323, 2 (former banshanyuan No. 12 Building 1 layer 1) Applicant before: Qin Yinlin |
|
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20081119 |