CN104262508A - Preparation technique of tinzaparin sodium - Google Patents
Preparation technique of tinzaparin sodium Download PDFInfo
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- CN104262508A CN104262508A CN201410480869.0A CN201410480869A CN104262508A CN 104262508 A CN104262508 A CN 104262508A CN 201410480869 A CN201410480869 A CN 201410480869A CN 104262508 A CN104262508 A CN 104262508A
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Abstract
The invention relates to a preparation technique of tinzaparin sodium. The technical scheme is as follows: the technique comprises the following steps: preparing a heparin sodium solution, preparing a heparin sodium enzyme solution, preparing a heparin sodium degradation solution, refining, freeze-drying and the like. The weight average molecular weight of the product is 5500-7500; the components with the molecular weight of less than 2000 account for not more than 10%, the components with the molecular weight of 2000-8000 account for 60-72%, and the components with the molecular weight of greater than 8000 account for 22-36%; and the anti-X activity is greater than or equal to 110 IU/mg. The technique has the advantages of abundant raw material sources, high yield, stable and reliable quality, high purity, simple technique and no discharge of three wastes, and is convenient to operate; and the product has the anticoagulant, antithrombotic, antitumor, anti-inflammatory, antiallergic and other effects, has obvious curative effect, can be used for preventing deep venous thrombosis after surgery, preventing venous thrombosis during hemodialysis and extracorporeal circulation and treating deep venous thrombosis, and has wide application range.
Description
Technical field
The present invention relates to a kind of spit of fland heparin sodium preparation technology.
Background technology
Spit of fland heparin sodium (Tinzaparin Sodium) low molecule sodium salt is developed by Aktiebolaget Leo (SE) Box 941, S-251 09 Helsingborg, Sweden of Denmark, and in June, 1996 listing, trade(brand)name Innohep.In Europe listing, in July, 2000 was through FDA approval listing in 1991 for spit of fland heparin sodium.The spit of fland heparin sodium global marketing volume of 2013 about 6.2 hundred million dollars, in Low molecular heparin series products before the sales volume single variety whole world four.Meanwhile, domesticly there is no this marketing drugs, there is great market potential.
The sodium salt of the CSSO3 that heparin system extracts in the intestinal mucosa of pig or ox, belongs to mucopolysaccharide class material.There is the linear chain structure of six Tang Huobatang repeating units, its molecular weight is between 3000 ~ 37000, molecular-weight average about 15000, heparin has the multiple pharmacologically actives such as anti-freezing, anti-inflammatory, antianaphylaxis, antiviral, anticancer, adjusting blood lipid, but life-time service then may produce some untoward reaction, as hemorrhage and induced platelet minimizing etc.Low molecular heparin (1owmolecular weight heparin, LMWH) is heparin fractionated or degraded and the less fragment of the molecular mass obtained.Over nearly 10 years.People find the research of Low molecular heparin, and its anti thrombotic action is better than heparin, have that bioavailability is high, Half-life in vivo is long, bleeding tendency is little, the features such as oral easy absorption.Because Low molecular heparin validity and security are all better than unfractionated heparin, become the focus of heparin class drug research at present.
The preparation method of Low molecular heparin LMWH mainly chemical cleavage method and enzymolysis process in the market, chemical cleavage method is mainly by β-null method, nitrous acid edman degradation Edman, hydrogen peroxide degradation method, acid degradation method and ray edman degradation Edman etc., react more violent in these method operating process, easily make the group of heparin end produce to destroy, cause the biological activity units change more or less of LMWH, thus likely cause the instability of whole product.And enzymolysis process is exactly by the low molecule heparin product needed for depolymerized heparin one-tenth under comparatively gentle condition.
Summary of the invention
Object of the present invention is exactly in view of the foregoing defects the prior art has, a kind of spit of fland heparin sodium preparation technology is provided, heparinase I is utilized to cut off heparin and the company key of Suleparoid (relative reactivity is about 3:1) between glucosamine and zero-sulfuric acid iduronic acid, depolymerized heparin is become disaccharide small molecules, thus obtain this product of spit of fland heparin.
Its technical scheme comprises following preparation process:
(1), heparin sodium aqua is prepared:
Refined heparin sodium is dissolved as the solution of 5% ~ 10% concentration, regulates material liquid pH value 7.0 ~ 8.0, temperature 25-30 DEG C;
(2) heparin sodium enzymolysis solution, is prepared:
Adding the heparinase of the 1.0%-3.0% of the refined heparin sodium liquor capacity of above-mentioned steps, is liquid, and enzymolysis time is 1.0 ~ 2.0h, intermittent stirring;
The ratio absorbance having reacted rear its feed liquid 232nm of on-line checkingi is 0.9 ~ 1.2, and rapidly feed liquid is warming up to 80.0 ~ 95.0 DEG C, 0.2 ~ 0.5h stops whole DeR;
(3) enzymolysis solution throw out, is prepared:
The liquid refrigerating completed by enzymolysis, adds enzymolysis solution 2 ~ 3 times of medicinal alcohols, fully stirs to obtain its throw out;
(4) fine work spit of fland heparin sodium, is prepared:
By above-mentioned throw out melt into 10% ~ 15% concentration, be further processed with the Sterile Filtration film of 0.22 micron, the finished product carry out lyophilize and obtain spit of fland heparin sodium.
Preferably, in step 3 of the present invention, be distributed in 5500-7500 by Liquid Detection main molecules amount, the component that molecular weight is less than 2000 is not more than 10%, and the component of 2000 ~ 8000, between 60% and 72%, is greater than the component of 8000 between 22% and 36%.
Compared with the prior art, advantage and the beneficial effect of present invention process are in the present invention:
1) utilize heparin sodium for raw material, by control degradation and uv-absorbing on-line monitoring, make DeR more controlled, be conducive to the generation reducing by product, realize the large-scale production of spit of fland heparin sodium;
2) present invention process is adopted, the weight-average molecular weight of products obtained therefrom is 5500-7500, the component that molecular weight is less than 2000 is not more than 10%, and the component of 2000 ~ 8000, between 60% and 72%, is greater than the component of 8000 between 22% and 36%, anti-Ⅹ Huo≤110IU/mg, anti-II a of anti-Ⅹ a/ is 1.5-2.5, and this product antithrombotic acitivity is better, and purity is high, impurity is few, and product is stablized.
Embodiment
Embodiment 1: add 900ml purified water to 1kg refined heparin sodium (tire 175usp/mg), until completely dissolved, feed temperature is adjusted to 27 DEG C, then with the sodium hydroxide that concentration is 6mol/L, material liquid pH value is adjusted to 7.3, accurate measuring heparin sodium aqua volume is 1000ml, add the heparinase of heparin sodium aqua volume 1%, i.e. 10ml, enzymolysis 2.0h.In reaction process, intermittent stirring, enzymolysis is complete, and the photoabsorption of its feed liquid 232nm of on-line checkingi is 0.9, rapidly feed liquid is warming up to 87 DEG C, and reaction 0.3h, stops whole DeR.The liquid refrigerating completed by enzymolysis, adds enzymolysis solution 2 ~ 3 times of medicinal alcohols, fully stirs to obtain its throw out;
Its molecular weight of Liquid Detection is: main molecules amount is 6150, the component that molecular weight is less than 2000 be 9.0%, 2000 ~ 8000 component be 67%, the component being greater than 8000 is 32%;
By the concentration of spit of fland heparin sodium throw out melt into 10%, feed liquid carries out Sterile Filtration by 0.22um filter membrane, filtrate lyophilize, obtains spit of fland heparin sodium 3.95 Kg.
Embodiment 2: add 2500ml purified water to 3kg refined heparin sodium (tire 187usp/mg), until completely dissolved, feed temperature is adjusted to 25 DEG C, then with the sodium hydroxide that concentration is 6mol/L, material liquid pH value is adjusted to 7.5, accurate measuring heparin sodium aqua volume is 2690ml, add the heparinase of heparin sodium aqua volume 1.5%, i.e. 40.35ml, enzymolysis 1.5h.Reaction process should monitor pH, temperature, intermittent stirring, and enzymolysis is complete, and the ratio photoabsorption that on-line ultraviolet detects its feed liquid 232nm is 1.0, rapidly feed liquid is warming up to 92 DEG C, and reaction 0.4h, stops whole DeR.The liquid refrigerating completed by enzymolysis, adds enzymolysis solution 2 ~ 3 times of medicinal alcohols, fully stirs to obtain its throw out;
Its molecular weight of Liquid Detection is: main molecules amount is 5930, the component that molecular weight is less than 2000 be 8.9%, 2000 ~ 8000 component be 68%, the component being greater than 8000 is 29%;
By the concentration of above-mentioned throw out melt into 10%, feed liquid carries out Sterile Filtration by 0.22um filter membrane, filtrate lyophilize, obtains spit of fland heparin sodium 4.15 Kg.
Embodiment 3: add 4800ml purified water to 5kg refined heparin sodium (tire 191usp/mg), until completely dissolved, feed temperature is adjusted to 30 DEG C, then with the sodium hydroxide that concentration is 6mol/L, material liquid pH value is adjusted to 8.0, accurate measuring heparin sodium aqua volume is 5000ml, add the heparinase of heparin sodium aqua volume 2%, i.e. 100.00ml, enzymolysis 1.0h.Reaction process should monitor pH, temperature, intermittent stirring, and enzymolysis is complete, and the photoabsorption that on-line ultraviolet detects its feed liquid 232nm is 1.1, rapidly feed liquid is warming up to 95 DEG C, and reaction 0.4h, stops whole DeR.The liquid refrigerating completed by enzymolysis, adds enzymolysis solution 2 ~ 3 times of medicinal alcohols, fully stirs to obtain its throw out;
Its molecular weight of Liquid Detection is: main molecules amount is 6200, the component that molecular weight is less than 2000 be 8.5%, 2000 ~ 8000 component be 69%, the component being greater than 8000 is 31%;
By the concentration of above-mentioned throw out melt into 10%, feed liquid carries out Sterile Filtration by 0.22um filter membrane, filtrate lyophilize, obtains spit of fland heparin sodium 4.21Kg.
Anti-Ⅹ Huo≤110IU/mg of the present invention; And abundant raw material source, productive rate are high, steady quality is reliable, purity is high, technique is simple, easy to operate, three-waste free discharge.Product spit of fland heparin sodium has anticoagulation, antithrombotic, antitumor, effect such as anti-inflammatory, antianaphylaxis, evident in efficacy.Can be used for prevention of surgical Post operation dvt to be formed, venothrombotic formation when preclude blood dialysis and extracorporeal circulation, treatment deep venous thrombosis.Can effectively prevent unstable coronary heart disease, use range is extensive.
Claims (2)
1. a spit of fland heparin sodium preparation technology, is characterized in that comprising following preparation process:
(1), heparin sodium aqua is prepared:
Refined heparin sodium is dissolved as the solution of 5% ~ 10% concentration, regulates material liquid pH value 7.0 ~ 8.0, temperature 25-30 DEG C;
(2) heparin sodium enzymolysis solution, is prepared:
Adding the heparinase of the 1.0%-3.0% of the refined heparin sodium liquor capacity of above-mentioned steps, is liquid, and enzymolysis time is 1.0 ~ 2.0h, intermittent stirring;
The ratio absorbance having reacted rear its feed liquid 232nm of on-line checkingi is 0.9 ~ 1.2, and rapidly feed liquid is warming up to 80.0 ~ 95.0 DEG C, 0.2 ~ 0.5h stops whole DeR;
(3) enzymolysis solution throw out, is prepared:
The liquid refrigerating completed by enzymolysis, adds enzymolysis solution 2 ~ 3 times of medicinal alcohols, fully stirs to obtain its throw out;
(4) fine work spit of fland heparin sodium, is prepared:
By above-mentioned throw out melt into 10% ~ 15% concentration, be further processed with the Sterile Filtration film of 0.22 micron, the finished product carry out lyophilize and obtain spit of fland heparin sodium.
2. spit of fland heparin sodium preparation technology according to claim 1, it is characterized in that: in step 3,5500-7500 are distributed in by Liquid Detection main molecules amount, the component that molecular weight is less than 2000 is not more than 10%, the component of 2000 ~ 8000, between 60% and 72%, is greater than the component of 8000 between 22% and 36%.
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| CN201410480869.0A CN104262508A (en) | 2014-09-19 | 2014-09-19 | Preparation technique of tinzaparin sodium |
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| CN201410480869.0A CN104262508A (en) | 2014-09-19 | 2014-09-19 | Preparation technique of tinzaparin sodium |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104774279A (en) * | 2015-03-26 | 2015-07-15 | 深圳市海普瑞药业股份有限公司 | Novel ultra low molecular weight heparin produced from enzyme |
| CN111073921A (en) * | 2019-12-30 | 2020-04-28 | 东营天东制药有限公司 | Preparation method of heparin sodium |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5106734A (en) * | 1986-04-30 | 1992-04-21 | Novo Nordisk A/S | Process of using light absorption to control enzymatic depolymerization of heparin to produce low molecular weight heparin |
| CN1421464A (en) * | 2002-11-29 | 2003-06-04 | 上海惠海生化制品厂 | Low molecular weight heparine sodium (calcium) and its prepn |
| CN102558393A (en) * | 2011-12-31 | 2012-07-11 | 河北常山生化药业股份有限公司 | Preparation process of dalteparin sodium |
-
2014
- 2014-09-19 CN CN201410480869.0A patent/CN104262508A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5106734A (en) * | 1986-04-30 | 1992-04-21 | Novo Nordisk A/S | Process of using light absorption to control enzymatic depolymerization of heparin to produce low molecular weight heparin |
| CN1421464A (en) * | 2002-11-29 | 2003-06-04 | 上海惠海生化制品厂 | Low molecular weight heparine sodium (calcium) and its prepn |
| CN102558393A (en) * | 2011-12-31 | 2012-07-11 | 河北常山生化药业股份有限公司 | Preparation process of dalteparin sodium |
Non-Patent Citations (1)
| Title |
|---|
| 刘亚梅等: "酶法制备低分子量肝素及其活性研究", 《药物生物技术》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104774279A (en) * | 2015-03-26 | 2015-07-15 | 深圳市海普瑞药业股份有限公司 | Novel ultra low molecular weight heparin produced from enzyme |
| CN111073921A (en) * | 2019-12-30 | 2020-04-28 | 东营天东制药有限公司 | Preparation method of heparin sodium |
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Application publication date: 20150107 |