CN103342761B - A kind of membrane sepn prepares Enoxaparin Sodium technique - Google Patents
A kind of membrane sepn prepares Enoxaparin Sodium technique Download PDFInfo
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- 229960005153 enoxaparin sodium Drugs 0.000 title claims abstract description 25
- CIJQTPFWFXOSEO-NDMITSJXSA-J tetrasodium;(2r,3r,4s)-2-[(2r,3s,4r,5r,6s)-5-acetamido-6-[(1r,2r,3r,4r)-4-[(2r,3s,4r,5r,6r)-5-acetamido-6-[(4r,5r,6r)-2-carboxylato-4,5-dihydroxy-6-[[(1r,3r,4r,5r)-3-hydroxy-4-(sulfonatoamino)-6,8-dioxabicyclo[3.2.1]octan-2-yl]oxy]oxan-3-yl]oxy-2-(hydroxy Chemical compound [Na+].[Na+].[Na+].[Na+].O([C@@H]1[C@@H](COS(O)(=O)=O)O[C@@H]([C@@H]([C@H]1O)NC(C)=O)O[C@@H]1C(C[C@H]([C@@H]([C@H]1O)O)O[C@@H]1[C@@H](CO)O[C@H](OC2C(O[C@@H](OC3[C@@H]([C@@H](NS([O-])(=O)=O)[C@@H]4OC[C@H]3O4)O)[C@H](O)[C@H]2O)C([O-])=O)[C@H](NC(C)=O)[C@H]1C)C([O-])=O)[C@@H]1OC(C([O-])=O)=C[C@H](O)[C@H]1O CIJQTPFWFXOSEO-NDMITSJXSA-J 0.000 title claims abstract description 25
- 239000012528 membrane Substances 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 12
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- 239000000047 product Substances 0.000 claims abstract description 28
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 claims abstract description 18
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- 238000000108 ultra-filtration Methods 0.000 claims abstract description 16
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- 238000009826 distribution Methods 0.000 claims abstract description 8
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
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- 238000003756 stirring Methods 0.000 claims description 21
- -1 heparinate compound Chemical class 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 16
- 238000001556 precipitation Methods 0.000 claims description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 229960002897 heparin Drugs 0.000 claims description 10
- 238000013019 agitation Methods 0.000 claims description 9
- 229960001950 benzethonium chloride Drugs 0.000 claims description 9
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical class OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims description 8
- UREZNYTWGJKWBI-UHFFFAOYSA-M benzethonium chloride Chemical compound [Cl-].C1=CC(C(C)(C)CC(C)(C)C)=CC=C1OCCOCC[N+](C)(C)CC1=CC=CC=C1 UREZNYTWGJKWBI-UHFFFAOYSA-M 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 239000001632 sodium acetate Substances 0.000 claims description 6
- 235000017281 sodium acetate Nutrition 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000003960 organic solvent Substances 0.000 claims description 4
- 239000000243 solution Substances 0.000 claims description 4
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 3
- 239000008186 active pharmaceutical agent Substances 0.000 claims description 3
- KCXMKQUNVWSEMD-UHFFFAOYSA-N benzyl chloride Chemical compound ClCC1=CC=CC=C1 KCXMKQUNVWSEMD-UHFFFAOYSA-N 0.000 claims description 3
- 229940073608 benzyl chloride Drugs 0.000 claims description 3
- 150000007942 carboxylates Chemical class 0.000 claims description 3
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- 230000000694 effects Effects 0.000 abstract description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 4
- 208000032843 Hemorrhage Diseases 0.000 abstract description 3
- 230000002785 anti-thrombosis Effects 0.000 abstract description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 abstract description 3
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- CVCQAQVBOPNTFI-AAONGDSNSA-N (3r,4r,5s,6r)-3-amino-6-(hydroxymethyl)oxane-2,4,5-triol;sulfuric acid Chemical compound OS(O)(=O)=O.N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O.N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O CVCQAQVBOPNTFI-AAONGDSNSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses a kind of membrane separation technique and prepare Enoxaparin Sodium technique.Heparin sodium realizes β-degraded, oxidative decoloration, filtering and impurity removing, filtrate again through uf processing through process such as quaternary ammonium salt salinization, benzyl esterification, alkaline bleach liquor degradations successively, obtains molecular-weight average and the satisfactory Enoxaparin Sodium product of range of molecular weight distributions.The present invention adopts the membrane ultrafiltration of different pore size to realize controlling the molecular weight and molecualr weight distribution of product, prepares high purity, highly active Enoxaparin Sodium product.The present invention takes hydrogen peroxide oxidation to decolour and activated carbon filtration removal of impurities, effectively eliminates the impurity in reaction system and significantly improves product colourity.Enoxaparin Sodium weight-average molecular weight prepared by the present invention is 3800 ~ 5000, molecular mass and distribution scope is all more satisfactory, the anti-F Ⅹ a/ anti-F II a ratio recorded is greater than 3.3, the principal element of hemorrhage risk anti-F II a activity is caused significantly to reduce, play the active raising relatively of anti-F Ⅹ a of antithrombotic effect, validity and the security advantages of product are obvious.
Description
Technical field
The present invention relates to the method utilizing membrane separation technique to prepare Enoxaparin Sodium, belong to technical field of bioengineering.
Technical background
The class long chain polysaccharides that heparin (heparin) is made up of glucosamine sulphate and hexuronic acid, is extensively present in the animal organ such as mucous membrane of small intestine, lung and tissue, and main separation from mammiferous mastocyte obtains at present.Heparin is by protein binding different from other, important biological activity can be played, there is the various biological functions such as anti-freezing, anti-inflammatory, antianaphylaxis, antiviral, anticancer, Adjust-blood lipid, it can activate antithrombin, accelerate the speed that antithrombin suppresses serine protease in coagulation cascade, thus be widely used as the anticoagulant medicine of one.But life-time service can produce many negative impacts, as side effects such as hemorrhage and induced platelet minimizing, osteoporosises.These shortcomings limit heparin application clinically.Therefore, people have developed novel anticoagulation medicine, low molecular weight heparin (LMWH).This kind of medicine has that chain is short, combining site is few, and bioavailability is high, and the advantages such as long half time, thus there is good medical prospect.
Enoxaparin Sodium is the one of Low molecular heparin, is mainly used in antithrombotic, anticoagulation, treatment acute myocardial infarction and unstable angina pectoris etc.Its anti-freezing complicated mechanism, indication is relatively more extensive, all has effect to the links of blood coagulation.Comprise anticoagulant proenzyme and change zymoplasm into, anticoagulant enzymic activity, obstruction conversion of fibrinogen is scleroproein, prevents platelet aggregation.Enoxaparin Sodium, by reducing LDL and VLDL, raises HDL, changes blood viscosity, and protection vascular endothelial cell, pre-preventing thrombosis and atherosclerosis, improve the effects such as coronary artery circulation.It is wide that Enoxaparin Sodium has clinical indication, and the advantages such as bioavailability is high, and Half-life in vivo is long, and side effect is little, its share of market reaches more than 2/3 of heparin class medicine.
What Enoxaparin Sodium produced main employing at present is β-elimination edman degradation Edman technique, the common problem that this processing method exists is that the target product molecular size range obtained is uneven, need to carry out classification to different molecular weight section, to reach medicinal requirements, and the traditional ethanol fractionation precipitation processing method step of widespread use is comparatively loaded down with trivial details, and produce a large amount of waste ethanol need recycling.
Summary of the invention
For prior art Problems existing, the present invention takes to carry out uf processing with the film of different pore size to liquid, intercepts and obtains molecular-weight average and range of molecular weight distributions satisfactory high reactivity Enoxaparin Sodium product.
For realizing the object of the invention, this membrane sepn is prepared Enoxaparin Sodium technique and is comprised the following steps:
A. quaternary ammonium salt salinization
By the heparin sodium aqueous solution and the benzyl rope chlorine aqueous solution with 1/1.5 ~ 2.5 volume ratio mix after react at 45 ~ 50 DEG C, react after 2 ~ 4 hours and throw out washing, oven dry are obtained heparin quaternary ammonium salt, described heparin sodium and benzethonium chloride mass concentration ratio are 1/2.5 ~ 3;
B. benzyl esterification
Heparin quaternary ammonium salt is dissolved into after in 5 ~ 7 quality DMF organic solvent doubly, after adding heparin quaternary ammonium salt 1 ~ 4 quality Benzyl Chloride doubly, at 25 ~ 35 DEG C, carries out esterification;
C. degrade
Esterification reaction product is added sodium hydroxide and reacts at 40 ~ 60 DEG C by above-mentioned esterification again after 20 ~ 30 hours, described sodium hydroxide concentration is 6 ~ 12% of esterification reaction product weight;
D. oxidative decoloration
After above-mentioned reaction 30 ~ 60min, degraded product is added the hydrogen peroxide oxidation decolouring of 30% concentration of 1 ~ 1.5%v/v reaction solution volume;
E. filtering and impurity removing
After adding gac with the consumption of 40 ~ 80g/L in above-mentioned reaction system, filter under 0.05 ~ 0.12MPa;
F. classification ultrafiltration
By filter membrane under 0.22 ~ 0.28MPa the ultrafiltration of the filtrate of filtering and impurity removing with MWCO8kDa aperture, to remove the heparin sodium degraded product of molecular weight higher than 8kDa, filtrate uses the ultrafiltration under 0.18 ~ 0.24MPa of the filter membrane in MWCO2kDa aperture again, obtains the Enoxaparin Sodium degraded product of molecular weight lower than 2kDa to retain.
Beneficial effect of the present invention: crucial being of this invention adopts the membrane ultrafiltration technology of different pore size to realize controlling the molecular weight and molecualr weight distribution scope of product, prepares high purity, highly active Enoxaparin Sodium product.Production technique of the present invention utilizes β-null method that general macromolecular heparin sodium is degraded to low molecular weight heparin sodium under given conditions.This invention takes hydrogen peroxide oxidation decolouring and the technique of activated carbon filtration removal of impurities, effectively eliminate the impurity in reaction system and significantly improve the colourity of product.
Accompanying drawing explanation
Fig. 1 is Enoxaparin Sodium production technological process of the present invention.
Embodiment
Embodiment 1: as shown in Figure 1,
A. hydrophilic reaction system prepares heparin-benzethonium chloride salt
In 30 liters of 30 ~ 40 DEG C of purified water, add 3kg heparin sodium (injection stage removes DS), stir and make it to dissolve completely; In 50 liters of 45 ~ 50 DEG C of purified water, add 8kg benzethonium chloride, stir and make it to dissolve completely.Then under agitation slowly joined in the benzethonium chloride solution of dissolving by the heparin sodium aqua of dissolving, after adding at 45 ~ 50 DEG C after Keep agitation 30 ~ 60min, by centrifugal for the throw out generated, abandoning supernatant, obtains heparinate compound.After 100 liters of purified water washing heparinate compounds, centrifugal abandoning supernatant, after washing centrifugal repetitive operation 3 ~ 4 times, opens Hotaircirculatingoven, by the heparinate compound after centrifugal at 50 ~ 60 DEG C dry 18 ~ 24 hours in an oven, obtain water content lower than 10% heparinate compound;
B. hydrophobic reactant system prepares heparin benzyl ester
Setting bath temperature 30 ~ 35 DEG C, dried heparinate compound is joined in 50 liters of dinethylformamide organic solvents to stir and make it to dissolve, add 10L Benzyl Chloride until completely dissolved, insulation esterification is stirred after 20 ~ 30 hours at 25 ~ 35 DEG C, add 200 liter of 3% ~ 10% sodium acetate ethanolic soln again to stir, then the sodium acetate ethanolic soln agitator treating throw out of 150 liter 3% ~ 10% is under agitation added, add a small amount of purified water dissolution precipitation thing again, after adding the sodium-chlor stirring and dissolving of liquor capacity 5% ~ 10% again, add 250 liter of 95% ethanol and stir precipitation, stop stirring latter standing 6 hours, repeat salt adding precipitation operation 2 times,
C. β under alkaline condition-elimination degraded
Above-mentioned carboxylate precipitation purified water is dissolved into the feed liquid of mass concentration 10%, during warming-in-water to 40 ~ 60 DEG C, after 220g sodium hydroxide is dissolved by purified water, add in feed liquid and start degraded, maintain liquid temperature 40 ~ 60 DEG C in degradation process, pH value is not less than 10.0, within every 10 minutes, survey a liquid temperature and pH value, after degraded 30 ~ 60min, logical cooling water temperature, adds hydrochloric acid and is neutralized to pH value 6.0 ~ 8.0, then alcohol precipitation after vacuum filtration;
D. Enoxaparin Sodium decolouring and removal of impurities
After above-mentioned degraded product being added purified water dissolving, add 1% ~ 1.5%(v/v) reaction liquid volume concentrations be 30% hydrogen peroxide rear oxidation 6 ~ 8 hours, reacting liquid temperature controls at 20 ~ 30 DEG C, and then in reaction system, adding Plate Filtration after gac with the consumption of 40 ~ 80g/L, filter pressure controls at 0.05 ~ 0.12MPa;
E. membrane ultrafiltration is adopted to carry out classification to different molecular weight component
By the filtrate after Plate Filtration after the membrane filtration in 0.2 μm of aperture, adopt the filter membrane of larger aperture (MWCO8kDa) in 0.22 ~ 0.28MPa, ultrafiltration at 20 ~ 35 DEG C, to remove the heparin sodium degraded product of most of molecular weight higher than 8kDa, again by the filtrate that obtains and then through the filter membrane of smaller aperture due (MWCO2kDa) in 0.18 ~ 0.24MPa, ultrafiltration at 20 ~ 35 DEG C, to retain the heparin sodium degraded product of most of molecular weight lower than 2kDa.Molecular-weight average and the satisfactory Enoxaparin Sodium product of the present invention of range of molecular weight distributions is obtained by after the essence filter successively of the concentrated trapped fluid obtained, lyophilize.The model that wherein two times of ultrafiltration all adopts Millipore Corp. to produce is the polysulfone membrane of 2KD.
Enoxaparin Sodium weight-average molecular weight prepared by the present invention is 3800 ~ 5000, the anti-F Ⅹ a/ anti-F II a ratio recorded is greater than 3.3, the principal element of hemorrhage risk anti-F II a activity is caused significantly to reduce, and the active raising relatively of anti-F Ⅹ a playing antithrombotic effect, validity and the security advantages of product are obvious.Table 1 lists the comparative result of Enoxaparin Sodium product prepared by the present invention and country and imported product standard.
Embodiment 2: the present embodiment difference from Example 1 is:
A. hydrophilic reaction system prepares heparin-benzethonium chloride salt
In 50 liters of 30 ~ 40 DEG C of purified water, add 5kg heparin sodium (injection stage removes DS), stir and make it to dissolve completely; In 85 liters of 45 ~ 50 DEG C of purified water, add 13kg benzethonium chloride, stir and make it to dissolve completely.Then under agitation slowly joined in the benzethonium chloride solution of dissolving by the heparin sodium aqua of dissolving, after adding at 45 ~ 50 DEG C after Keep agitation 30 ~ 60min, by centrifugal for the throw out generated, abandoning supernatant, obtains heparinate compound.After 100 liters of purified water washing heparinate compounds, centrifugal abandoning supernatant, after washing centrifugal repetitive operation 3 ~ 4 times, opens Hotaircirculatingoven, by the heparinate compound after centrifugal at 50 ~ 60 DEG C dry 18 ~ 24 hours in an oven, obtain water content lower than 10% heparinate compound;
B. hydrophobic reactant system prepares heparin benzyl ester
Setting bath temperature 30 ~ 35 DEG C, dried heparinate compound is joined 105 liters of N, stir in dinethylformamide organic solvent and make it to dissolve, add 16 liters of Benzyl Chlorides until completely dissolved, insulation esterification is stirred after 20 ~ 30 hours at 25 ~ 35 DEG C, add 500 liter of 3% ~ 10% sodium acetate ethanolic soln again to stir, then the sodium acetate ethanolic soln agitator treating throw out of 300 liter 3% ~ 10% is under agitation added, add a small amount of purified water dissolution precipitation thing again, then after adding the sodium-chlor stirring and dissolving of liquor capacity 5% ~ 10%, add 400 liter of 95% ethanol and stir precipitation, stop stirring latter standing 6 hours, repeat salt adding precipitation operation 2 times.
C. β under alkaline condition-elimination degraded
Above-mentioned carboxylate precipitation purified water is dissolved into the feed liquid of mass concentration 10%, during warming-in-water to 40 ~ 60 DEG C, after the sodium hydroxide of 6% ~ 12% is dissolved by purified water, add in feed liquid and start degraded, maintain liquid temperature 40 ~ 60 DEG C in degradation process, pH value is not less than 10.0, within every 10 minutes, survey a liquid temperature and pH value, after degraded 30 ~ 60min, logical cooling water temperature, adds hydrochloric acid and is neutralized to alcohol precipitation after pH value 6.0 ~ 8.0 final vacuum suction filtration;
D. Enoxaparin Sodium decolouring and removal of impurities
Above-mentioned degraded product is added purified water to dissolve, reacting liquid temperature controls at 20 ~ 30 DEG C, add 1% ~ 1.5%(v/v) reaction liquid volume concentrations be 30% hydrogen peroxide oxidation 6 ~ 8 hours, in reaction system, add Plate Filtration after gac with the consumption of 40 ~ 80g/L again, filter pressure controls at 0.05 ~ 0.12MPa;
E. membrane ultrafiltration is adopted to carry out classification to different molecular weight component
By the filtrate after Plate Filtration after the membrane filtration in 0.2 μm of aperture, adopt the filter membrane of larger aperture (MWCO8kDa) in 0.22 ~ 0.28MPa, ultrafiltration at 20 ~ 35 DEG C, to remove the heparin sodium degraded product of most of molecular weight higher than 8kDa, again by the filtrate that obtains and then through the filter membrane of smaller aperture due (MWCO2kDa) in 0.18 ~ 0.24MPa, ultrafiltration at 20 ~ 35 DEG C, to retain the heparin sodium degraded product of most of molecular weight lower than 2kDa.Molecular-weight average and the satisfactory Enoxaparin Sodium product of the present invention of range of molecular weight distributions is obtained by after the essence filter successively of the concentrated trapped fluid obtained, lyophilize.The model that wherein two times of ultrafiltration all adopts Millipore Corp. to produce is the polysulfone membrane of 2KD.
Claims (1)
1. membrane sepn prepares an Enoxaparin Sodium technique, it is characterized in that it comprises the following steps,
A. hydrophilic reaction system prepares heparin-benzethonium chloride salt
In 30 liters of 30 ~ 40 DEG C of purified water, add 3kg injection stage remove DS heparin sodium, stir and make it to dissolve completely, 8kg benzethonium chloride is added in 50 liters of 45 ~ 50 DEG C of purified water, stirring makes it to dissolve completely, then the heparin sodium aqua of dissolving is under agitation slowly joined in the benzethonium chloride solution of dissolving, after adding at 45 ~ 50 DEG C after Keep agitation 30 ~ 60min, by centrifugal for the throw out generated, abandoning supernatant, obtain heparinate compound, after 100 liters of purified water washing heparinate compounds, centrifugal abandoning supernatant, after washing centrifugal repetitive operation 3 ~ 4 times, open Hotaircirculatingoven, by the heparinate compound after centrifugal at 50 ~ 60 DEG C dry 18 ~ 24 hours in an oven, obtain water content lower than 10% heparinate compound,
B. hydrophobic reactant system prepares heparin benzyl ester
Setting bath temperature 30 ~ 35 DEG C, dried heparinate compound is joined 50 liters of N, stir in dinethylformamide organic solvent and make it to dissolve, add 10L Benzyl Chloride until completely dissolved, insulation esterification is stirred after 20 ~ 30 hours at 25 ~ 35 DEG C, add 200 liter of 3% ~ 10% sodium acetate ethanolic soln again to stir, then the sodium acetate ethanolic soln agitator treating throw out of 150 liter 3% ~ 10% is under agitation added, add a small amount of purified water dissolution precipitation thing again, after adding the sodium-chlor stirring and dissolving of liquor capacity 5% ~ 10% again, add 250 liter of 95% ethanol and stir precipitation, stop stirring latter standing 6 hours, repeat salt adding precipitation operation 2 times,
C. β under alkaline condition-elimination degraded
Above-mentioned carboxylate precipitation purified water is dissolved into the feed liquid of mass concentration 10%, during warming-in-water to 40 ~ 60 DEG C, after 220g sodium hydroxide is dissolved by purified water, add in feed liquid and start degraded, maintain liquid temperature 40 ~ 60 DEG C in degradation process, pH value is not less than 10.0, within every 10 minutes, survey a liquid temperature and pH value, after degraded 30 ~ 60min, logical cooling water temperature, adds hydrochloric acid and is neutralized to pH value 6.0 ~ 8.0, then alcohol precipitation after vacuum filtration;
D. Enoxaparin Sodium decolouring and removal of impurities
After above-mentioned degraded product being added purified water dissolving, add the hydrogen peroxide rear oxidation 6 ~ 8 hours that 1% ~ 1.5%v/v reaction liquid volume concentrations is 30%, reacting liquid temperature controls at 20 ~ 30 DEG C, and then in reaction system, adding Plate Filtration after gac with the consumption of 40 ~ 80g/L, filter pressure controls at 0.05 ~ 0.12MPa;
E. membrane ultrafiltration is adopted to carry out classification to different molecular weight component
By the filtrate after Plate Filtration after the membrane filtration in 0.2 μm of aperture, adopt the filter membrane of aperture MWCO8kDa at 0.22 ~ 0.28MPa, ultrafiltration at 20 ~ 35 DEG C, to remove the heparin sodium degraded product of most of molecular weight higher than 8kDa, again by the filtrate that obtains and then through the filter membrane of aperture MWCO2kDa at 0.18 ~ 0.24MPa, ultrafiltration at 20 ~ 35 DEG C, to retain the heparin sodium degraded product of most of molecular weight lower than 2kDa, by the essence filter successively of the concentrated trapped fluid obtained, molecular-weight average and the satisfactory Enoxaparin Sodium product of range of molecular weight distributions is obtained after lyophilize.
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| CN103936889A (en) * | 2014-03-19 | 2014-07-23 | 苏州英诺凯生物医药科技有限公司 | Method for purification of enoxaparin by tangential flow filtration |
| 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 |
| CN106977627A (en) * | 2017-05-16 | 2017-07-25 | 苏州二叶制药有限公司 | A kind of Enoxaparin production method of sodium |
| AU2018385557B2 (en) | 2017-12-11 | 2024-01-04 | Biological E Limited | Process for the preparation of low molecular weight heparin |
| CN109293800B (en) * | 2018-08-16 | 2022-02-25 | 山东万邦赛诺康生化制药股份有限公司 | Method for removing benzyl chloride taste in production process of heparin benzyl ester |
| 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 |
| AR118776A1 (en) * | 2019-04-26 | 2021-10-27 | Farm Rovi Lab Sa | PROCEDURE FOR OBTAINING LOW MOLECULAR WEIGHT HEPARINS BY TANGENTIAL FLOW FILTRATION |
| WO2020216981A1 (en) | 2019-04-26 | 2020-10-29 | Laboratorios Farmacéuticos Rovi, S.A. | Method for obtaining low-molecular-weight heparins by means of tangential flow filtration |
| CN115043959A (en) * | 2022-05-25 | 2022-09-13 | 湖北亿诺瑞生物制药有限公司 | Preparation method of high-yield enoxaparin sodium |
| CN116284499B (en) * | 2022-07-28 | 2024-07-23 | 河北常山生化药业股份有限公司 | Preparation method of sheep-derived low-molecular heparin sodium |
| CN117946294A (en) * | 2023-11-13 | 2024-04-30 | 河北常山凯库得生物技术有限公司 | Preparation method of energy-saving and environment-friendly enoxaparin sodium |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101165071A (en) * | 2006-10-20 | 2008-04-23 | 江苏江山制药有限公司 | Clexane and preparation method thereof |
| CN102050888A (en) * | 2010-12-13 | 2011-05-11 | 河北常山生化药业股份有限公司 | Method for preparing enoxaparin sodium |
| CN102558392A (en) * | 2010-12-14 | 2012-07-11 | 王芃 | Preparation method of high-FXa-resistant low-FIIa-resistant low-molecular heparin sodium |
| CN102603925A (en) * | 2012-03-21 | 2012-07-25 | 东营天东生化工业有限公司 | Method for directly producing enoxaparin sodium from crude product heparin sodium |
| CN102633908A (en) * | 2012-05-02 | 2012-08-15 | 雷晓刚 | Method for preparing high-quality LMW (low molecular weight) heparins |
-
2013
- 2013-07-15 CN CN201310295978.0A patent/CN103342761B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101165071A (en) * | 2006-10-20 | 2008-04-23 | 江苏江山制药有限公司 | Clexane and preparation method thereof |
| CN102050888A (en) * | 2010-12-13 | 2011-05-11 | 河北常山生化药业股份有限公司 | Method for preparing enoxaparin sodium |
| CN102558392A (en) * | 2010-12-14 | 2012-07-11 | 王芃 | Preparation method of high-FXa-resistant low-FIIa-resistant low-molecular heparin sodium |
| CN102603925A (en) * | 2012-03-21 | 2012-07-25 | 东营天东生化工业有限公司 | Method for directly producing enoxaparin sodium from crude product heparin sodium |
| CN102633908A (en) * | 2012-05-02 | 2012-08-15 | 雷晓刚 | Method for preparing high-quality LMW (low molecular weight) heparins |
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