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Preparation method of chymotrypsin and medicine composition for improving stability of chymotrypsin
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A chymotrypsin and protein technology, which is applied in the field of biopharmaceuticals, can solve the problems of expensive affinity chromatography medium, large amount of medium used, and few reusable times, and achieves the effect of low cost and large market competitiveness.
Inactive Publication Date: 2016-03-16
QINGDAO KANGYUAN PHARMA
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Problems solved by technology
[0003] Chinese patent CN1944641A discloses a preparation method of chymotrypsin and a pharmaceutical composition for improving the stability of chymotrypsin, and adopts an affinity chromatography process to produce high-purity chymotrypsin from prochymotrypsin, but as analyzed in Chinese patent CN1013025B, there are also some shortcomings , the most important thing is that the affinity chromatography medium is expensive, the number of reusable times is small, and the amount of medium used is large, which is not suitable for large-scale production
Although the Chinese patent CN1013025B has solved the deficiency of affinity chromatography, it can also obtain high specific activity chymotrypsin, but similarly, compared with the traditional process, the production cost is too high
Method used
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Effect test
Embodiment 1
[0041] 1. Crude production operation of chymotrypsin:
[0042] (1) Weigh 4kg of freshly frozen bovine pancreas, immediately remove the fatty connective tissue with tweezers and scissors, mince it, and weigh it;
[0043] (2) Put the minced material in a plastic bucket and soak it with 6L of 0.125mol / L sulfuric acid solution for 22 hours;
[0044] (3) Filter the impregnated matter, and impregnate the filter residue with 4L of 0.125mol / L sulfuric acid solution for 1 hour, and discard the filter residue after filtering;
[0045] (4) Combine the two impregnating solutions, measure a volume of 11.4L, put it in a plastic bucket, add 2.76kg of solidammoniumsulfate to reach 40% saturation, and let it stand for 12 hours;
[0046] (5) Filtrate, keep the filtrate, measure the volume of 10.2L, and discard the solid;
[0047] (6) Add 2.1kg of solidammoniumsulfate to the filtrate to reach 70% saturation, and let it stand for 12 hours;
[0048] (7) Filter, retain the solid, weigh 116g,...
Embodiment 2
[0074] 1. Crude production operation of chymotrypsin:
[0075] (1) Weigh 4kg of freshly frozen bovine pancreas, immediately remove the fatty connective tissue with tweezers and scissors, mince it, and weigh it;
[0076] (2) Put the minced material in a plastic bucket and soak it with 10L of 0.125mol / L sulfuric acid solution for 28 hours;
[0077] (3) Filter the impregnation, and impregnate the filter residue with 8L of 0.125mol / L sulfuric acid solution for 2.5 hours, and discard the filter residue after filtering;
[0078] (4) Combine the two impregnating solutions, measure a volume of 11.5L, put it in a plastic bucket, add 2.78kg of solid ammoniumsulfate to reach 40% saturation, and let it stand for 15 hours;
[0079] (5) Filtrate, keep the filtrate, measure a volume of 10.3L, and discard the solid;
[0080] (6) Add 2.1kg of solid ammonium sulfate to the filtrate to reach 70% saturation, and let it stand for 15 hours;
[0081] (7) Filter, retain the solid, weigh 118g, and...
Embodiment 3
[0107] Take 880,000 units of chymotrypsin prepared in Example 2, weigh 15g of mannitol and 4g of sodiumbicarbonate, add 500ml of water for injection to dissolve, adjust the pH value to 5.8 after mixing, then add water for injection to 1000ml, polyethersulfone super Membrane sterile filtration, packed in 1000 treated vials, freeze-dried and capped.
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Abstract
The invention discloses a preparation method of chymotrypsin and a medicine composition for improving the stability of chymotrypsin. The preparation method includes the following processing steps: 1, mincing raw materials and extracting protein; 2, conducting graded salting out and obtaining a chymotrypsin crude product; 3, feeding the crude product and conducting multiple crystallization; 4, conducting activation; 5, conducting salting out; 6, conducting dialysis; 7, conducting freeze-drying. By means of the preparation method of chymotrypsin and the medicine composition for improving the stability of chymotrypsin, a scaled production process is established, and the preparation method and the medicine composition have the advantages of being simple, low in cost and the like; the boutique titer of the prepared chymotrypsin reaches as high as 1200 unit / mg or more and meets the requirements of 'Chinese Pharmacopoeia' (Version 2010), and huge market competitiveness is achieved.
Description
technical field [0001] The invention belongs to the field of biopharmaceuticals and relates to a biochemical separation technology of medicinal chymotrypsin. Specifically, techniques such as salting out, multiple crystallization combined with dialysis are used to extract chymotrypsin. Background technique [0002] Chymotrypsin is a protease isolated and purified from bovine pancreas or porcine pancreas. Its pharmacological and clinical functions mainly include two aspects: 1. Anti-inflammation: it has good curative effect on various inflammations, inflammatory edema, hematoma, ulcer and thrombus. 2. Anti-cancer: It can promote the penetration of anti-cancer drugs into the lesion, promote the effect of chemotherapy drugs, and the effect is stronger when used locally in large doses. Large-dose use can treat a variety of tumors, such as breast cancer, cervical cancer, gastric cancer, etc., without any toxic side effects or adverse reactions. [0003] Chinese patent CN1944641...
Claims
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Application Information
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