DK200401720A - New mixts. of heparin-based sulphated polysaccharide(s) - are used to prevent post-operative and arterial thrombosis and has longer half-life than known heparin prod - Google Patents

New mixts. of heparin-based sulphated polysaccharide(s) - are used to prevent post-operative and arterial thrombosis and has longer half-life than known heparin prod Download PDF

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DK200401720A
DK200401720A DK200401720A DKPA200401720A DK200401720A DK 200401720 A DK200401720 A DK 200401720A DK 200401720 A DK200401720 A DK 200401720A DK PA200401720 A DKPA200401720 A DK PA200401720A DK 200401720 A DK200401720 A DK 200401720A
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Denmark
Prior art keywords
heparin
process according
mixts
prod
operative
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DK200401720A
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Danish (da)
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Debrie Roger
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Aventis Pharma Sa
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Priority claimed from FR9008013A external-priority patent/FR2663639B1/en
Application filed by Aventis Pharma Sa filed Critical Aventis Pharma Sa
Priority to DK200401720A priority Critical patent/DK200401720A/en
Publication of DK200401720A publication Critical patent/DK200401720A/en

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  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Mixed sulphated polysaccharides (A) having the basic structure of heparin-forming polysaccharides are characterised by a wt. average mol. wt. (Mw) below that of heparin; 9-20% chains have mol. wt. below 2000 and 5-20% chains have mol. wt. over 8000. The ratio Mw:number average mol. wt. (Mn) is 1.3-1.6. - Pref. (A) contains less than 2% dermatan sulphate and has Mw 3500-5500. The polysaccharide chain has at one end 2-O-sulpho-4-enopyranosuronic acid.

Description

Patentkrav 1. Blandinger af sulfaterede polysaccharider med samme generelle struktur som de polysaccharider, der indgår i heparin, kendetegnet ved, at de har lavere gennemsnitsmolekylvægt efter vægt end heparin, at de indeholder mellem 9 og 20% kæder med en molekylvægt på under 2000 dalton og mellem 5 og 20% kæder med en molekylvægt på over 8000 dalton, og at forholdet mellem gennemsnitsmolekylvægten efter vægt og gennemsnitsmolekylvægten efter antal er på mellem 1,3 og 1,6. 2. Blandinger ifølge krav 1, kendetegnet ved, at de indeholder mindre end 2% dermatansulfat. 3. Blandinger ifølge krav 1 eller 2, kendetegne t ved, at de har en gennemsnitsmolekylvægt efter vægt på mellem ca. 3500 og ca. 5500 dalton. 4. Blandinger ifølge ethvert af kravene 1-3, kendetegnet ved, at kæderne af sulfaterede polysaccharider indeholder en 2-0-sulfo-4-enopyranosuronsyre ved en af kædeenderne. 5. Fremgangsmåde til fremstilling af blandinger af sulfaterede polysaccharider ifølge ethvert af kravene 1-4, kendetegnet ved, at der gennemføres de følgende trin: {a) i et første trin omdannes et heparin i vandholdigt medium til et salt deraf ved hjælp af et kvaternært ammoniumsalt med lang kæde, (b) i et andet trin forestres det fremkomne salt til en ester med en forestringsgrad mellem 9,5 og 14%, og (c) i et tredje trin depolymeriseres den fremkomne ester med en forestringsgrad mellem 9,5 og 14%. 6. Fremgangsmåde ifølge krav 5, kendeteg net ved, at det andet trin gennemføres i et chloreret organisk opløsningsmiddel i nærværelse af en chlorforbin-delse. 7. Fremgangsmåde ifølge krav 6, kendeteg net ved, at chlorforbindelsen er benzylchlorid, og at opløsningsmidlet er chloroform eller methylenchlorid. 8. Fremgangsmåde ifølge ethvert af kravene 5-7, kendetegnet ved, at forestringen gennemføres ved blanding af 1 vægtdel heparinsalt med ca. 1 volumendel af en chlorforbindelse i 3-5 volumendele af et chloreret organisk opløsningsmiddel ved en temperatur på mellem 25 og 45°C, fortrinsvis mellem 30 og 40°C. 9. Fremgangsmåde ifølge krav 5, kendetegnet ved, at det tredje trin gennemføres ved, at esteren behandles med en stærk base i vandig opløsning. 10. Fremgangsmåde ifølge krav 9, kendeteg net ved, at vægt forholdet mellem base og ester er mellem 0,05 og 2, fortrinsvis mellem 0,08 og 0,15. 11. Fremgangsmåde ifølge krav 9, kendeteg net ved, at vægtforholdet mellem vand og ester er mellem 15 og 30. 12. Fremgangsmåde ifølge krav 9, kendete g- n e t ved, at mediets temperatur indstilles på en værdi på mellem 50 og 70°C, fortrinsvis mellem 55 og 65°C, og at reaktionen gennemføres i løbet af 30 minutter til 3 timer, fortrinsvis i løbet af 1-2 timer. 13. Fremgangsmåde ifølge ethvert af kravene 5-12, kendetegnet ved, at man i et indledende trin udfælder udgangsheparinet ved hjælp af en alkohol. 14. Anvendelse af blandinger af sulfaterede polysac-charider ifølge ethvert af kravene 1-4 til fremstilling af terapeutiske præparater. 15. Anvendelse ifølge krav 14 til fremstilling af terapeutiske præparater beregnet til anvendelse til forhindring af venethromboser efter operationer. 16. Fremgangsmåde ifølge krav 5, kendetegnet ved, at urenheder fjernes fra blandingerne. 17. Fremgangsmåde ifølge krav 16, kendetegnet ved, at urenheder valgt blandt nucleinsyrer, poly-peptider og polysaccharider, især chondroitinsulfater, heparan- og dermatansulfat, fjernes. 18. Fremgangsmåde ifølge krav 17, kendetegnet ved, at dermantansulfatet fjernes til et restindhold på under 2%. 19. Fremgangsmåde ifølge ethvert af de foregående krav 5 og 16-18, kendetegnet ved, at der fremstilles blandinger med en gennemsnitsmolekylvægt mellem ca. 3500 dalton og ca. 5500 dalton. 20. Fremgangsmåde ifølge krav 5, kendetegnet ved, at i det første trin omdannes heparinet til benzethoniumsaltet. 21. Fremgangsmåde ifølge krav 5, kendetegne t ved, at i det andet trin gennemføres forestringen med et chlorderivat, især benzylchlorid. 22. Fremgangsmåde ifølge krav 5, kendetegnet ved, at i det andet trin gennemføres forestringen med et organisk opløsningsmiddel. 23. Fremgangsmåde ifølge krav 5, kendetegnet ved, i det andet trin reguleres mængderne af de forskellige reagenser og temperaturen og reaktionstiden. 24. Fremgangsmåde ifølge krav 5, kendetegnet ved, at i det andet trin af forestringen dannes en aromatisk ester som heparinester. 25. Fremgangsmåde ifølge krav 5, kendetegnet ved, at esteren, der er dannet i det andet trin, udvindes ved udfældning med alkohol, især methanol, i nærværelse af natriumacetat. 26. Fremgangsmåde ifølge krav 5, kendetegnet ved, at forestringsgraden af heparinesteren reguleres, især ved højeffektiv væskechromatografi i væskefase. 27. Fremgangsmåde ifølge krav 5, kendetegnet ved, at i det tredje trin gennemføres depolymerisa-tionen med en stærk base i vandig opløsning, især med natriumhydroxid . 28. Terapeutisk præparat på basis af blandinger af sulfaterede polysaccharider med samme generelle strukturer som de polysaccharider, der indgår i heparin, og som har en lavere gennemsnitsmolekylvægt efter vægt end heparin, som indeholder mellem 9 og 20% kæder med en molekylvægt på under 2000 dalton og mellem 5 og 20% kæder med en molekylvægt på over 8000 dalton, og hvis forhold mellem gennemsnitsmolekylvægten efter vægt og gennemsnitsmolekylvægten efter antal ligger mellem 1,3 og 1,6. 29. Præparat ifølge krav 28, som er et antithrombotisk præparat.Patent claims 1. Mixtures of sulfated polysaccharides having the same general structure as the polysaccharides included in heparin, characterized in that they have lower average molecular weight by weight than heparin, containing between 9 and 20% chains having a molecular weight of less than 2000 daltons and between 5 and 20% chains with a molecular weight greater than 8000 daltons, and that the ratio between the average molecular weight by weight and the average molecular weight by number is between 1.3 and 1.6. Mixtures according to claim 1, characterized in that they contain less than 2% dermatan sulfate. Mixtures according to claim 1 or 2, characterized in that they have an average molecular weight by weight of between approx. 3500 and approx. 5500 daltons. Mixtures according to any one of claims 1-3, characterized in that the chains of sulfated polysaccharides contain a 2-O-sulfo-4-enopyranosuronic acid at one of the chain ends. Process for preparing mixtures of sulfated polysaccharides according to any one of claims 1-4, characterized in that the following steps are carried out: (a) in a first step a heparin in aqueous medium is converted to a salt thereof by means of a quaternary (b) in a second step, the resulting salt is esterified to an ester with a degree of esterification between 9.5 and 14%, and (c) in a third step, the resulting ester is depolymerized with a degree of esterification between 9.5 and 14.5%. 14%. Process according to claim 5, characterized in that the second step is carried out in a chlorinated organic solvent in the presence of a chlorine compound. Process according to claim 6, characterized in that the chlorine compound is benzyl chloride and that the solvent is chloroform or methylene chloride. Process according to any one of claims 5-7, characterized in that the esterification is carried out by mixing 1 part by weight of heparin salt with approx. 1 part by volume of a chlorine compound in 3-5 parts by volume of a chlorinated organic solvent at a temperature between 25 and 45 ° C, preferably between 30 and 40 ° C. Process according to claim 5, characterized in that the third step is carried out by treating the ester with a strong base in aqueous solution. Process according to claim 9, characterized in that the weight ratio of base to ester is between 0.05 and 2, preferably between 0.08 and 0.15. Process according to claim 9, characterized in that the weight ratio of water to ester is between 15 and 30. A process according to claim 9, characterized in that the temperature of the medium is adjusted to a value between 50 and 70 ° C. , preferably between 55 and 65 ° C, and the reaction is carried out over 30 minutes to 3 hours, preferably in 1-2 hours. Process according to any one of claims 5-12, characterized in that, in an initial step, the starting heparin is precipitated by an alcohol. Use of mixtures of sulfated polysaccharides according to any one of claims 1-4 for the preparation of therapeutic compositions. Use according to claim 14 for the manufacture of therapeutic compositions intended for use in preventing venous thrombosis after surgery. Process according to claim 5, characterized in that impurities are removed from the mixtures. Process according to claim 16, characterized in that impurities selected from nucleic acids, polypeptides and polysaccharides, especially chondroitin sulphates, heparan and dermatan sulphate, are removed. Process according to claim 17, characterized in that the dermantane sulphate is removed to a residual content of less than 2%. Process according to any one of the preceding claims 5 and 16-18, characterized in that mixtures having an average molecular weight of between approx. 3500 daltons and approx. 5500 daltons. Process according to claim 5, characterized in that in the first step the heparin is converted to the benzethonium salt. Process according to claim 5, characterized in that in the second step the esterification is carried out with a chlorine derivative, especially benzyl chloride. Process according to claim 5, characterized in that the esterification is carried out with an organic solvent. The process according to claim 5, characterized in that in the second step, the amounts of the various reagents and the temperature and reaction time are controlled. Process according to claim 5, characterized in that in the second step of the esterification an aromatic ester is formed as a heparin ester. Process according to claim 5, characterized in that the ester formed in the second step is recovered by precipitation with alcohol, especially methanol, in the presence of sodium acetate. Process according to claim 5, characterized in that the degree of esterification of the heparin ester is controlled, especially by high-efficiency liquid phase chromatography. Process according to claim 5, characterized in that in the third step the depolymerization is carried out with a strong base in aqueous solution, especially with sodium hydroxide. 28. Therapeutic composition based on mixtures of sulfated polysaccharides having the same general structures as the polysaccharides included in heparin and having a lower average molecular weight by weight than heparin containing between 9 and 20% chains with a molecular weight below 2000 daltons and between 5 and 20% chains with a molecular weight greater than 8000 daltons and whose ratio between the average molecular weight by weight and the average molecular weight by number is between 1.3 and 1.6. The composition of claim 28, which is an antithrombotic preparation.

DK200401720A 1990-06-26 2004-11-09 New mixts. of heparin-based sulphated polysaccharide(s) - are used to prevent post-operative and arterial thrombosis and has longer half-life than known heparin prod DK200401720A (en)

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Application Number Priority Date Filing Date Title
DK200401720A DK200401720A (en) 1990-06-26 2004-11-09 New mixts. of heparin-based sulphated polysaccharide(s) - are used to prevent post-operative and arterial thrombosis and has longer half-life than known heparin prod

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR9008013A FR2663639B1 (en) 1990-06-26 1990-06-26 LOW MOLECULAR WEIGHT POLYSACCHARIDE BLENDS PROCESS FOR PREPARATION AND USE.
DK199101243A DK176524B1 (en) 1990-06-26 1991-06-25 Mixtures of sulfated polysaccharides and their preparation and use
DK200401720A DK200401720A (en) 1990-06-26 2004-11-09 New mixts. of heparin-based sulphated polysaccharide(s) - are used to prevent post-operative and arterial thrombosis and has longer half-life than known heparin prod

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