CN105440163A - Method for preparing and purifying enoxaparin sodium - Google Patents
Method for preparing and purifying enoxaparin sodium Download PDFInfo
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- CN105440163A CN105440163A CN201510882503.0A CN201510882503A CN105440163A CN 105440163 A CN105440163 A CN 105440163A CN 201510882503 A CN201510882503 A CN 201510882503A CN 105440163 A CN105440163 A CN 105440163A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof
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Abstract
The invention discloses a method for preparing and purifying enoxaparin sodium. After a crude enoxaparin sodium product is subjected to centrifugal separation and alcohol precipitation impurity removal, the finished enoxaparin sodium product meeting pharmacopeia standards can be obtained. The method is high in yield and easy and convenient to implement, and the product is good in clarity and capable of meeting requirements of the pharmaceutical industry.
Description
Technical field
The invention belongs to field of biological pharmacy, relate to a kind of production and purification process of Enoxaparin Sodium particularly.
Background technology
Heparin has been the choice drug of prevention and therapy thrombus class medicine since the eighties in last century always.Low molecular heparin is made up by specific chemical chop and purifies and separates of unfractionated heparin, and due to its determined curative effect, the advantages such as side effect is little and measurable, progressively instead of the market position of traditional heparin.
Enoxaparin Sodium (EnoxaparinSodium) belongs to Low molecular heparin, and principal character is have a 4-alkene pyranose aldehydic acid structure in molecule.This product anti thrombotic action is strong, hemorrhage risk is little, is one of current anticoagulation, antithrombotic primary treatment medicine.
Enoxaparin Sodium obtains by carrying out alkaline hydrolysis poly-to the benzyl ester derivative of chitling mucous membrane heparin, its basic step is: with pig mucous membrane heparin for starting raw material, through the preparation of heparin quaternary ammonium salt, the preparation of heparin benzyl ester, heparin benzyl ester carried out to alkaline hydrolysis and gather, with acid neutralization, alcohol precipitation, refining, decolouring, dehydrate, obtain Enoxaparin Sodium finished product.Wherein, in acid and, the Enoxaparin Sodium intermediate that obtains after alcohol precipitation, its clarity is often very poor, if not in addition polishing purification, just cannot reach the requirement of clarity in pharmacopeia.
Existing purification process is mainly divided into following several: (1) is by Enoxaparin Sodium activated carbon decolorizing, filter through macroporous resin, filtrate freeze-drying obtains purifying Enoxaparin Sodium, the method decolorizing effect is better, but shortcoming has a large amount of products to be adsorbed in gac and resin, and yield only can reach about 60%; (2) Enoxaparin Sodium sodium hydroxide is adjusted pH, by hydrogen peroxide for decoloration, freeze-drying after sodium-chlor and washing with alcohol, obtain purifying Enoxaparin Sodium, the method needs to use the extremely strong hydrogen peroxide of oxidisability, while removing pigment, easily cause the generation of other by products, and environmental pollution is comparatively large, is difficult to reach environmental impact assessment requirement; (3) Enoxaparin Sodium is passed through hydrophobic chromatography post and anionite-exchange resin successively, after nanofiltration desalination, precipitation, the dry Enoxaparin Sodium obtaining purifying, the method needs too much time chromatography column, and product loss is very large; (4) Enoxaparin Sodium is passed through filtration, micro-filtration and ultra-filtration equipment successively, obtain purifying Enoxaparin Sodium after freeze-drying, the method product loss is less, but the working pressure of this classification ultrafiltration is comparatively large, is not suitable for industrially applying; (5) Enoxaparin Sodium sodium chloride solution and methyl alcohol are processed repeatedly, the Enoxaparin Sodium that final acquisition clarity is up to standard, the method is easy and simple to handle, easily industrially realize, but its purification effect is poor, and the number of times repeatedly processed is difficult to accurately determine, is unfavorable for the foundation of production standard at every turn.
There is many drawbacks in the market outlook wide due to Enoxaparin Sodium and existing purification technique, developing a kind of method that can be applicable to that industrial production uses, fast and convenient purifying Enoxaparin Sodium just becomes problem demanding prompt solution.
Summary of the invention
The object of this invention is to provide the method for a kind of yield raising Enoxaparin Sodium purity high, easy and simple to handle.
A method for Enoxaparin Sodium preparation and purifying, is characterized in that, comprise the following steps:
(1) by Enoxaparin Sodium crude product, put into sodium chloride solution and dissolve, obtain Enoxaparin Sodium crude product solution, filter;
(2) by step (1) gained filtrate through centrifugation, obtain supernatant liquor and throw out;
(3) ethanol or methyl alcohol are joined in the supernatant liquor obtained in step (2), stir and be precipitated thing after leaving standstill;
(4) throw out purified water is dissolved into the solution of 10wt ~ 15wt%, after 0.1 μm of membrane filtration, spraying dry obtains Enoxaparin Sodium finished product.
In step (1), the concentration of sodium chloride solution is 5-15%, and the weight/volume of Enoxaparin Sodium crude product and sodium chloride solution is 1:5-10.
In step (2), the rotating speed of centrifugation is 1000 ~ 3000r/min, and centrifugation time is 10min ~ 60min.
In step (3), the volume ratio of ethanol or methyl alcohol and supernatant liquor is 1-5:1, and churning time is 1 ~ 3h, and time of repose is 3-10h.
Research finds, impurity affect Enoxaparin Sodium clarity is mainly derived from high molecular weight protein in heparin sodium and micromolecular oligosaccharides, and these impurity are in the preparation process of Enoxaparin Sodium, and aldehyde radical warp " Maillard reaction " in its molecule can produce coloring matter.The present invention first adopts that high-speed low temperature is centrifugal eliminates macromolecular albumen, then by small molecular weight impurities such as alcohol deposition method removing oligosaccharides, thus reach the object of purifying Enoxaparin Sodium.
Compared with existing purification process, the method for the invention yield can reach more than 90%, and product clarity is good, and purification process is easy, is applicable to very much pharmaceutical industry and produces.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Embodiment 1: the preparation of Enoxaparin Sodium crude product
Step (1): the preparation of heparin benzethonium chloride salt
In retort, drop into heparin sodium 1kg and purified water 20L, stir about 20 minutes, dissolve completely, transfer in cancellation tank.In retort, drop into benzethonium chloride 2.5kg and purified water 20L, stir about 20 minutes, dissolve completely, by benzethonium chloride solution suction cancellation tank, stirring reaction 30 minutes.Supplementary purified water 60L is added in cancellation tank, continue stirring 10 minutes, leave standstill 30 minutes.Filtered by reaction mixture, filter cake purified water pulls an oar 3 times (50kg, 50kg, 60kg).Collect filter cake, under 60 DEG C ± 2 DEG C hot water, vacuum tightness are greater than 0.08MPa, vacuum-drying 10 hours, obtains heparin benzethonium chloride salt 2.1kg.
Step (2): the preparation of heparin benzyl ester
In the esterification tank of clean dried, drop into heparin benzethonium chloride salt 2.1kg and methylene dichloride 20L, under whipped state, be warming up to 35 DEG C ± 1 DEG C, insulation reaction 1 hour.Drop into Benzyl Chloride 9kg, continue insulation reaction 25 hours.Reaction is finished, and adds sodium-acetate-methanol solution, finishes and continues reaction 30 minutes.Reacting liquid filtering, filter cake methyl alcohol is pulled an oar 2 times, then washs once.Collect filter cake, at room temperature vacuum-drying 10 hours, obtains heparin benzyl ester 1.3kg.
Step (3): the degraded of heparin benzyl ester
In degraded tank, drop into heparin benzyl ester 1.3kg and purified water 10L, under whipped state, be warming up to 60 DEG C ± 2 DEG C.Sodium hydroxide 150g and purified water 1L is dropped into, stirring and dissolving, temperature control to 60 DEG C ± 2 DEG C in alkali-prepared tank.By in the sodium hydroxide solution suction degraded tank in alkali-prepared tank, insulation reaction 2 hours.Reaction is finished, and is cooled to room temperature, drips 2N hcl acidifying, regulates reaction solution pH to 6 ~ 7, continues stirring 5 minutes, filters.Add sodium-chlor 0.5kg in filtrate, stir 30 minutes, dissolve completely and transfer in setting tank, add 5L methyl alcohol, finish, continue stirring 5 minutes, filter, collect filter cake, dry Enoxaparin Sodium crude product 1.1kg.
The purifying of embodiment 2 Enoxaparin Sodium
Enoxaparin Sodium crude product 1.1kg embodiment 1 prepared puts into the sodium chloride solution stirring and dissolving of 5L10%, obtains Enoxaparin Sodium crude product solution, filters and obtains filtrate.By filtrate 30min in the whizzer of 2000r/min, obtain supernatant liquor and throw out.10L ethanol is joined in supernatant liquor, leaves standstill 3h after stirring 1h, obtain throw out.Throw out purified water is dissolved into the solution of 10wt ~ 15wt%, after 0.1 μm of membrane filtration, spraying dry obtains Enoxaparin Sodium finished product 1.02kg, purification yield 92.7%.
The purifying of embodiment 3 Enoxaparin Sodium
Enoxaparin Sodium crude product 1.1kg embodiment 1 prepared puts into the sodium chloride solution stirring and dissolving of 10L5%, obtains Enoxaparin Sodium crude product solution, filters and obtains filtrate.By filtrate 30min in the whizzer of 2000r/min, obtain supernatant liquor and throw out.10L methyl alcohol is joined in supernatant liquor, leaves standstill 5h after stirring 1.5h, obtain throw out.Throw out purified water is dissolved into the solution of 10wt ~ 15wt%, after 0.1 μm of membrane filtration, spraying dry obtains Enoxaparin Sodium finished product 1.01kg, purification yield 91.8%.
Embodiment 4 inspection after construction
According to European Pharmacopoeia 8.0(EP8.0) method recorded detects the Enoxaparin Sodium finished product that the present invention prepares: weight-average molecular weight: 4350-4500
Be less than the molecular weight of 2000: 14%-18%
2000-8000 molecular weight: 72%-77%
Anti-Xa tires: 106-120IU/mg
Anti-IIa tires: 25-29IU/mg
Anti-Xa with the ratio of anti-IIa is: 3.5-4.0
pH：6.5-7.0
231nm absorbancy: 17-18
Phenylcarbinol remains: <0.05%
Sodium content: 11.5%-12.5%
Clarity: be shallower than No. 1 turbidity standard
Above index all reaches the standard of EP8.0, especially clarity one, far above record in EP8.0 not higher than the requirement of No. 6 turbidity standards.
The above is only preferred embodiment of the present invention, and be not restriction the present invention being made to other form, any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the Equivalent embodiments of equivalent variations.But everyly do not depart from technical solution of the present invention content, any simple modification, equivalent variations and the remodeling done above embodiment according to technical spirit of the present invention, still belong to the protection domain of technical solution of the present invention.
Claims (6)
1. a method for Enoxaparin Sodium preparation and purifying, its feature comprises the steps:
(1) Enoxaparin Sodium crude product is put into sodium chloride solution to dissolve, obtain Enoxaparin Sodium crude product solution, filter;
(2) by step (1) gained filtrate through centrifugation, obtain supernatant liquor and throw out;
(3) ethanol or methyl alcohol are joined in the supernatant liquor obtained in step (2), stir and be precipitated thing after leaving standstill;
(4) throw out purified water is dissolved into the solution of 10wt ~ 15wt%, after 0.1 μm of membrane filtration, spraying dry obtains Enoxaparin Sodium finished product.
2. the method for a kind of Enoxaparin Sodium preparation according to claim 1 and purifying, Enoxaparin Sodium crude product preparation method is wherein:
(1) preparation of heparin benzethonium chloride salt: drop into heparin sodium 1kg and purified water 20L in retort, stir about 20 minutes, dissolves completely, transfers in cancellation tank; In retort, drop into benzethonium chloride 2.5kg and purified water 20L, stir about 20 minutes, dissolve completely, by benzethonium chloride solution suction cancellation tank, stirring reaction 30 minutes; Supplementary purified water 60L is added in cancellation tank, continue stirring 10 minutes, leave standstill 30 minutes; Filtered by reaction mixture, filter cake purified water is pulled an oar 3 times, collects filter cake, and under 60 DEG C ± 2 DEG C hot water, vacuum tightness are greater than 0.08MPa, vacuum-drying 10 hours, obtains heparin benzethonium chloride salt 2.1kg;
(2) preparation of heparin benzyl ester: drop into heparin benzethonium chloride salt 2.1kg and methylene dichloride 20L in the esterification tank of clean dried, is warming up to 35 DEG C ± 1 DEG C under whipped state, insulation reaction 1 hour, drop into Benzyl Chloride 9kg, continue insulation reaction 25 hours, reaction is finished, add sodium-acetate-methanol solution, finish and continue reaction 30 minutes, reacting liquid filtering, filter cake methyl alcohol is pulled an oar 2 times, wash again once, collect filter cake, at room temperature vacuum-drying 10 hours, obtains heparin benzyl ester 1.3kg;
(3) degraded of heparin benzyl ester: drop into heparin benzyl ester 1.3kg and purified water 10L in degraded tank, be warming up to 60 DEG C ± 2 DEG C under whipped state; Sodium hydroxide 150g and purified water 1L is dropped into, stirring and dissolving, temperature control to 60 DEG C ± 2 DEG C in alkali-prepared tank; By in the sodium hydroxide solution suction degraded tank in alkali-prepared tank, insulation reaction 2 hours, reaction is finished, be cooled to room temperature, drip 2N hcl acidifying, regulate reaction solution pH to 6 ~ 7, continue stirring 5 minutes, filter, in filtrate, add sodium-chlor 0.5kg, stir 30 minutes, dissolve completely and transfer in setting tank, add 5L methyl alcohol, finish, continue stirring 5 minutes, filter, collect filter cake, dry Enoxaparin Sodium crude product 1.1kg.
3. the method for a kind of Enoxaparin Sodium preparation according to claim 1 and purifying, is characterized in that: in step (1), the concentration of sodium chloride solution is 5-15%, and the weight of Enoxaparin Sodium crude product and sodium chloride solution and volume ratio are 1:5-10.
4. the method for a kind of Enoxaparin Sodium preparation according to claim 1 and purifying, is characterized in that: in step (2), the rotating speed of centrifugation is 1000 ~ 3000r/min, and centrifugation time is 10min ~ 60min.
5. the method for a kind of Enoxaparin Sodium preparation according to claim 1 and purifying, is characterized in that: in step (3), the volume ratio of ethanol or methyl alcohol and supernatant liquor is 1-5:1, and churning time is 1 ~ 3h, and time of repose is 3-10h.
6. a kind of Enoxaparin Sodium preparation according to claim and the method for purifying, the feature of the refined heparin sodium produced is: weight-average molecular weight: 4350-4500; Be less than the molecular weight of 2000: 14%-18%; 2000-8000 molecular weight: 72%-77%; Anti-Xa tires: 106-120IU/mg; Anti-IIa tires: 25-29IU/mg; Anti-Xa is 3.5-4.0 with the ratio of anti-IIa; PH:6.5-7.0231nm; Absorbancy: 17-18; Phenylcarbinol remains: <0.05%; Sodium content: 11.5%-12.5%; Clarity: be shallower than No. 1 turbidity standard.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510882503.0A CN105440163A (en) | 2015-12-07 | 2015-12-07 | Method for preparing and purifying enoxaparin sodium |
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| CN201510882503.0A CN105440163A (en) | 2015-12-07 | 2015-12-07 | Method for preparing and purifying enoxaparin sodium |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106432547A (en) * | 2016-09-14 | 2017-02-22 | 苏州天马精细化学品股份有限公司 | Method for preparing enoxaparin sodium through heparin benzyl ester |
| CN109467621A (en) * | 2017-09-08 | 2019-03-15 | 山阳县恒瑞肉制品有限公司 | A kind of production of Enoxaparin Sodium and purification process |
| CN109666086A (en) * | 2018-11-05 | 2019-04-23 | 上海宝维医药技术有限公司 | A kind of preparation method and applications of High-purity heparin quaternary ammonium salt |
-
2015
- 2015-12-07 CN CN201510882503.0A patent/CN105440163A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106432547A (en) * | 2016-09-14 | 2017-02-22 | 苏州天马精细化学品股份有限公司 | Method for preparing enoxaparin sodium through heparin benzyl ester |
| CN109467621A (en) * | 2017-09-08 | 2019-03-15 | 山阳县恒瑞肉制品有限公司 | A kind of production of Enoxaparin Sodium and purification process |
| CN109666086A (en) * | 2018-11-05 | 2019-04-23 | 上海宝维医药技术有限公司 | A kind of preparation method and applications of High-purity heparin quaternary ammonium salt |
| CN109666086B (en) * | 2018-11-05 | 2020-12-22 | 上海宝维医药技术有限公司 | Preparation method and application of high-purity heparin quaternary ammonium salt |
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