Background technology
Dalteparin sodium is low molecular sodium salt, and it is mainly used in treatment, unstable coronary artery disease treatment and the prevention and the relevant thrombosis of performing the operation of pre-anticoagulant in the hemodialysis and deep venous thrombosis.Heparin (heparin) extensively is present in a kind of glycosaminoglycan in the tissue such as animal organ and mucous membrane of small intestine, lung, have anti-freezing, anti-inflammatory, antianaphylaxis, antiviral, anticancer, regulate the multiple pharmacologically active such as blood fat.
Low molecular heparin uses as anticoagulation except possessing the whole pharmacological actions of unfractionated heparin, usually short-term medication, and therapeutic index is high.U.S. FDA has been ratified the prophylactic of dalteparin sodium after as low level limbs displacement technique.
The method of the at present product of domestic production Low molecular heparin class degraded mainly contains the methods such as Sodium Nitrite edman degradation Edman, hydrogen peroxide degradation method, enzyme liberating method.But hydrogen peroxide degradation may be introduced foreign matter, and quality is not easy control; Enzyme liberating product end has unsaturated group, poor stability, and cost is high; On the removal of impurities decoloring method, present domestic mainly contain potassium permanganate and hydrogen peroxide secondary oxidation method or hydrogen peroxide oxidation process under condition of different pH.The shortcoming of potassium permanganate and hydrogen peroxide secondary oxidation method be the Manganse Dioxide that produces difficulty leach, and Manganse Dioxide has adsorption to portioned product, and product recovery rate is reduced; Technical at purifying products, the domestic methods such as gel chromatography, ultrafiltration that mainly contain although the quality product that gel chromatography obtains is higher, be difficult to large-scale production, and cost are higher.
Summary of the invention
The present invention aims to provide a kind of dalteparin sodium production technique, and DALT and the quality standard thereof prepared with this technique, the defective that exists to overcome prior art.
For realizing the object of the invention, dalteparin sodium production technique of the present invention comprises the following steps:
1., preparation heparin sodium aqua
After in heparin sodium, adding 5~8 quality purified water doubly, stir at normal temperatures dissolving after 4~6 hours;
2., preparation depolymerized heparin liquid
After the pH value 2~3 of the heparin sodium aqua that 1. obtains with the hydrochloric acid regulating step, adding weight is the Sodium Nitrite of heparin sodium 1.5%~2.5%, then at 10~30 ℃ of lower stirring reaction 2~5h, obtains depolymerized heparin liquid after leaving standstill 20~24h again;
3., preparation reduced liquid
Regulate the pH value 9~11 of above-mentioned depolymerized heparin liquid with sodium hydroxide solution after, add 0.6%~1.0% times of heparin sodium weight sodium borohydride reduction 10~16h, and then regulate pH value to 6.5~7.0 with sodium hydroxide solution, obtain reduced liquid;
4., preparation precipitate reduction thing
In above-mentioned reduced liquid, add the ethanol that 2~4 volumes are doubly measured, collect after fully stirring and obtain the precipitate reduction thing;
5., preparation dalteparin sodium crude product
After in the precipitate reduction thing of collecting, adding the pure water stirring and dissolving, the hydrogen peroxide that adding 0.2%~1.0% volume is doubly measured is at 25~35 ℃ of lower oxidation 10~24h, and then adding the ethanol that 2~4 volumes are doubly measured, the collecting precipitation thing obtains the dalteparin sodium crude product after fully stirring;
6., preparation dalteparin sodium elaboration
The dalteparin sodium crude product of above-mentioned collection precipitation being added purified water be dissolved into 1%~2%(w/w) solution, is that 3000 and 8000 ultra-filtration membrane carries out ultrafiltration with the molecular weight cutoff value respectively, obtains dalteparin sodium elaboration solution after Sterile Filtration again;
7., freeze-drying
The dalteparin sodium solution that above-mentioned steps is made namely obtains the dalteparin sodium finished product-25~-35 ℃ of lower freeze-drying.
The relevant comparative data with the domestic and imported standard of product of the present invention
The technical progress that the present invention obtains:
1, owing to adopts production technique of the present invention, with application of membrane separation technology in separating the different molecular weight section, the solvent precipitation classification and the expensive gel molecular sieve method that fall behind have been changed, having obtained anti-Xa tires 〉=130IU/mg, specific activity is 2.0~3.0, and the molecular weight distribution of main ingredient is the dalteparin sodium product of 5500~6500D.
2, the dalteparin sodium product that production technique acquisition molecular-weight average of the present invention is 5500~6500D can effectively prevent venous thromboembolism, can be used for preventing general surgery, orthopedics, before the art of Neurological Surgery and the thrombosis of postoperative, can effectively prevent the venous thromboembolism of Ischemic Apoplexy Patients, can greatly reduce the risk that suffers stroke, what can prevent effectively that blood extracorporeal circulation from causing solidifies, and can effectively prevent unstable coronary heart disease, can effectively solve be prone to behind the unfraction heparin life-time service hemorrhage, osteoporosis, induced platelet minimizings etc. have broad application prospects.
3, the present invention and existing preparation process of dalteparin sodium are relatively, productive rate is up to 85%~95%, and weight-average molecular weight is 5500~6500D, and peak molecular weight is 3500~6000, molecular weight is not more than 13% less than 3000 component, and molecular weight is not more than 15% greater than 8000 component; Anti-Xa activity 〉=130IU/mg, quality reaches the quality standard of European Pharmacopoeia regulation.The structure of product compound reducing end is the N.F,USP MANNITOL of dehydration, and has abundant, high a, stable and reliable product quality of productive rate of raw material sources, purity is high, and cost is low, adopts the Sodium Nitrite degraded so that technique is simple, easy to operate, production process is controlled easily, steady quality, reaction temperature and, product color is better, without characteristics such as " three wastes " dischargings.The effect that the product dalteparin sodium has the effect such as anticoagulation, antithrombotic, antitumor, anti-inflammatory, antianaphylaxis and regulates blood fat, evident in efficacy, use range is extensive, can be used as the raw material of the products such as injection, capsule, ointment, is convenient to scale production.
Embodiment
Embodiment 1: as shown in Figure 1, and at first:
1., take by weighing respectively behind 3Kg heparin sodium, 45g Sodium Nitrite, the 30g sodium borohydride for subsequent use;
2., add the 3Kg heparin sodium in 15~24L purified water retort after, stir at normal temperatures that dissolving obtains heparin sodium aqua after 4~6 hours;
3., regulate the pH=2 of above-mentioned heparin sodium aqua~3 with hydrochloric acid after, under whipped state, add the 45g Sodium Nitrite that takes by weighing, then 10~30 ℃ of insulations, react 2~5h under stirring, obtain depolymerized heparin liquid after leaving standstill again 20~24h;
4., regulate the pH value 9~11 of above-mentioned depolymerized heparin liquid with sodium hydroxide solution after, add 30g sodium borohydride reduction 10~16h, react complete after again with sodium hydroxide solution adjusting pH value to 6.5~7.0, obtain reduced liquid;
5., in above-mentioned reduced liquid, add the ethanol that 2~4 volumes are doubly measured, collection precipitate reduction thing after fully stirring;
6., in the precipitate reduction thing of collecting, add the pure water stirring and dissolving after, add 0.2%~1.0% volume hydrogen peroxide doubly at 25~35 ℃ of lower oxidation 10~24h, add again the ethanol that 2~4 volumes doubly measure after oxidation is complete and precipitate, collect after fully stirring and obtain dalteparin sodium throw out crude product;
7., above-mentioned dalteparin sodium crude product precipitation is added pure water 150L~200L, be dissolved into the solution of 1~2% concentration after, be respectively 3000 and 8000 ultra-filtration membrane with the molecular weight cutoff value and carry out ultrafiltration, after the degerming processing, obtain dalteparin sodium solution again;
8., the dalteparin sodium solution that above-mentioned steps is made obtains dalteparin sodium finished product 2.7Kg after-25~-35 ℃ of lower freeze-drying.
Embodiment 2: the present embodiment difference from Example 1 is that heparin sodium is 10Kg, and Sodium Nitrite is 150g, and sodium borohydride is 100g, obtains dalteparin sodium finished product 9Kg after the freeze-drying.
Embodiment 3: the present embodiment difference from Example 1 is that heparin sodium is 30Kg, and Sodium Nitrite is 450g, and sodium borohydride is 300g, obtains dalteparin sodium finished product 27Kg after the freeze-drying.