CN100402085C - Production of high-concentration thymic peptide solution and large-specification thymic peptide preparation - Google Patents
Production of high-concentration thymic peptide solution and large-specification thymic peptide preparation Download PDFInfo
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- CN100402085C CN100402085C CNB2004100092480A CN200410009248A CN100402085C CN 100402085 C CN100402085 C CN 100402085C CN B2004100092480 A CNB2004100092480 A CN B2004100092480A CN 200410009248 A CN200410009248 A CN 200410009248A CN 100402085 C CN100402085 C CN 100402085C
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Abstract
The present invention discloses a thymic peptide solution with high concentration, a preparation method thereof and a large-specification preparation prepared from the thymic peptide solution as raw medical material. In the present invention, a thymic peptide solution with low concentration is concentrated by a physical refrigeration technique to obtain a thymic peptide solution with high concentration. The method is characterized in that the method operated at low temperature prevents the bioactive ingredients of the product from being damaged; microorganisms are difficult in propagation in the state of low temperature so as to guarantee the hygienic requirement of the product; the content of thymic peptides in the concentrated thymic peptide solution can reach more than 50 mg/ml. The thymic peptide solution with high concentration is used as raw medical material for preparing thymopeptide preparations with the large specifications of 50 mg and 100 mg for injection.
Description
Technical field
The present invention relates to the preparation method of thymosin, more particularly, relate to the preparation method of high concentration thymus peptide solution.
The invention still further relates to big specification Thymopeptide by the inventive method preparation.
Background technology
Thymosin is an immunomodulator.Have the effect of adjusting and enhancing human body cell immunologic function, can impel T-lymphocyte maturation.Be mainly used in various constitutionales of treatment or Secondary cases T cell defect disease, some autoimmune disease, the disease that various cellular immune functions are low and the auxiliary treatment of tumor. comprising: 1. various hepatitis gravis, chronic active hepatitis, chronic persistent hepatitis and liver cirrhosis etc.; 2. herpes zoster, genital herpes, condyloma acuminatum etc.; 3. bronchitis, bronchial asthma, pulmonary tuberculosis are prevented upper respiratory tract infection etc.; 4. various malignant tumor early stages and chemotherapy, radiotherapy is share and is used; 5. lupus erythematosus, rheumatic and atrophic diseases, ankylosing spondylitis, Guillain Barre syndrome etc.; 6. aplastic anemia, leukemia, thrombocytopenia etc.; 7. viral keratitis, viral conjunctivitis, allergic rhinitis etc.; 8. senile senilism, climacteric syndrome etc.; 9. multiple furuncle and phyma and skin of face acne etc., psoriasis, lichen planus, squamous cell carcinoma and epithelium seborrheic keratosis etc.; 10. child's innate immunity defective disease etc.Using method is subcutaneous or intramuscular injection: 1 10～20mg, 1 time on the one.Intravenous drip: a 20～200mg is dissolved in 500ml 0.9% sodium chloride injection or 5% glucose injection, 1 time on the one.
Thymopeptide is owing to determined curative effect, and side effect is little, and using dosage has the trend of increasing.The dosage that is used for the treatment of at present diseases such as malignant tumor, hepatitis gravis, acquired immune deficiency syndrome (AIDS) clinically can reach 200mg, in the treatment scheme of atypical pneumonia, the using dosage of thymosin reached 300mg, and 1 time on the one, thereby heavy dose of Thymopeptide that uses has obtained clinical verification.On the other hand,, can only prepare the low specification preparation of 5mg, 10mg and 20mg, be difficult under low capacity, prepare the big specification preparation of 50mg and 100mg because of the thymus peptide solution content lower (less than 10mg/ml) that general extraction process obtains; And the preparation of low specification has a lot of shortcomings for heavy dose is used, and for example once uses many, many to open, dissolve, and not only during operating cost, and pollutes easily; On the other hand, the increase of drug packing material not only causes the waste of resource, and brings burden to environmental protection.
The most effectual way that improves thymus peptide solution content is concentrated.Method for concentration commonly used has distillation under vacuum, spray drying method, hyperfiltration, freeze-drying and freeze concentration method etc.After deliberation and contrast, distillation under vacuum and spray drying method need easily cause the bioactive loss of thymosin through pyroprocess; Hyperfiltration will also can influence the biological activity of thymosin through high-pressure process; Freeze-drying is difficult to satisfy large-scale production; Freeze concentration method can be avoided above-mentioned unfavorable conditions only, is suitable for the low content thymus peptide solution is concentrated.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of high concentration thymus peptide solution, this method not only can make the thymus peptide solution of thymosin content greater than 50mg/ml, and the product bioactive ingredients is not damaged, microorganism is difficult for breeding under low temperature state, has guaranteed the hygienic requirement of product.
Another object of the present invention is to provide a kind of Thymopeptide of big specification, comprises the preparation and the lyophilized formulations of solution form, and wherein the thymus peptide content can reach more than the 50mg/ml in the solution, and the thymosin content of unit formulation is 50～100mg in the lyophilized formulations.
The present invention discloses a kind of preparation method of high concentration thymus peptide solution, may further comprise the steps:
(1) extraction of thymus peptide solution: get thymus fresh or that quick-freezing is preserved, remove impurity and clean; The thymus of handling well is added water be prepared into homogenate; With the homogenate multigelation; Heating freeze thawing liquid is removed unsettled foreign protein, again through centrifugal, mistake leaching supernatant; The component of supernatant below ultrafiltration molecular cut off 10000 dalton obtains the thymus peptide solution of low concentration;
(2) concentrating of low concentration thymus peptide solution: adopt the physics concentration technique to prepare the high concentration thymus peptide solution low concentration thymus peptide solution that obtains in the step (1).
Described thymus comprises calf thymus or porcine thymus.
The content of thymosin is 5～10mg/ml in the low concentration thymus peptide solution that obtains in the step (1), and the content of thymosin is 50～60mg/ml in the high concentration thymus peptide solution that obtains in the step (2).
With the homogenate multigelation, be that homogenate is refrigerated to deep layer rapidly under-25～-35 ℃ of temperature in the step (1), in 20～30 ℃ water-bath, melt then, and repeatedly for several times.
The physics concentration technique that adopts in the step (2) comprises progressive freeze concentration method and suspension crystallization freeze concentration method, the preferred suspension crystallization freeze concentration method that adopts among the present invention.
Described suspension crystallization freeze concentration method carries out under-5～5 ℃ temperature and normal pressure, and its principle as shown in Figure 1.Among Fig. 1, abscissa is represented the concentration of solution, and vertical coordinate is represented the temperature of solution.Curve D ABE is the freezing point line of solution, and D is the freezing point of pure water, and E is an eutectic point.From the freezing point curve as seen, the freezing point of solution in the finite concentration scope descends along with the increase of concentration.A certain initial concentration is the weak solution of X1, lowers the temperature from temperature A1 point, and when temperature was reduced to freezing point line A point, if there be not " ice is planted " in the solution, then solution can't freeze.But when temperature continued to drop to the C point and becomes over-cooled solution, over-cooled solution was extremely unstable, in case be subjected to external interference (as vibrations), will produce ice crystal in the solution, and grew up and become the great achievement ice pellets.At this moment, along with the generation of ice crystal and ice pellets, the concentration of solution increases and is the ice pellets in the solution to be filtered out X2, can reach concentrated purpose.The solution of freeze concentration must be rarer, and its concentration must be less than eutectic point concentration.
The present invention also discloses a kind of high concentration thymus peptide solution that is made by the described preparation method of claim 1, it is characterized in that, wherein the content of thymosin is 50～60mg/ml.
It is the big specification preparation that crude drug makes that the present invention also discloses a kind of high concentration thymus peptide solution that made by preparation method of the present invention or of the present invention, and it comprises described high concentration thymus peptide solution and the pharmaceutically acceptable carrier and/or the solvent for the treatment of effective dose.
The high concentration thymus peptide solution that the present invention is made adds that pharmaceutically acceptable excipient can be made into the thymosin lyophilized injectable powder of big specification, thymosin in its unit formulation reaches 50～100mg, the unit formulation here refers generally to a preparation packing, for example an antibiotic bottle.Lyophilized formulations can supply the injection preparation of intramuscular injection, intravenous injection and intravenous drip.
The excipient that uses comprises mannitol or dextran, and its consumption is 3%～10% weight.
Method of the present invention is easy, can concentrate the thymus peptide solution of low content, makes the big specification Thymopeptide of content greater than 50mg/ml.Because this method is operated at low temperatures, has fully kept the biological activity of product, and can suppress microbial reproduction, thymus peptide solution for preparing and preparation meet the requirement of the national drug standards, have satisfied clinical needs.
Description of drawings
Fig. 1 is the schematic diagram of suspension crystallization freeze concentration method;
Fig. 2 is a thymus peptide solution preparation technology flow chart;
Fig. 3 is a suspension crystallization freeze concentration equipment sketch map;
Fig. 4 is an injection Thymopeptide process chart;
Fig. 5 is a thymus peptide solution component high-efficient liquid phase chromatogram.
The specific embodiment
The calf thymus 100Kg that gets the quick-freezing preservation puts in the special-purpose tank and melts, and extracts fat and adventitia, gets net weight 85Kg.The thymus of plucking cleans 3 times with drinking water earlier, and reuse water for injection cleans 3 times.After the meat grinder rubbing, add cold water for injection 85L, with colloid mill homogenate 2min.Homogenate put-30 ℃ freezing 48 hours, take out back 20 ℃ of water-baths and melt, be heated to 90 ℃ rapidly and kept 3 minutes, speed is chilled to room temperature.Centrifugal 20 minutes of 3000rpm gets supernatant liquid filtering, the 10000 daltonian ultrafilter ultrafiltration of filtrate reuse molecular cut off, and aseptic collection permeate is thymus peptide solution.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 5.9mg/ml, Thymosin alpha
1Account for 1.6% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
The calf thymus 100Kg that gets the quick-freezing preservation puts in the special-purpose tank and melts, and extracts fat and adventitia, gets net weight 80Kg.The thymus of plucking cleans 3 times with drinking water earlier, and reuse water for injection cleans 3 times.After the meat grinder rubbing, add cold water for injection 60L, with colloid mill homogenate 2min.Homogenate put-30 ℃ freezing 24 hours, take out 25 ℃ of water-baths next day and melt, 3 times repeatedly, be heated to 80 ℃ rapidly after melting for the third time and kept 10 minutes, speed is chilled to room temperature again.Centrifugal 20 minutes of 3000rpm gets supernatant liquid filtering, the 10000 daltonian ultrafilter ultrafiltration of filtrate reuse molecular cut off, and aseptic collection permeate is thymus peptide solution.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 7.7mg/ml, Thymosin alpha
1Account for 2.1% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
The calf thymus 100Kg that gets the quick-freezing preservation puts in the special-purpose tank and melts, and extracts fat and adventitia, gets net weight 83Kg.The thymus of plucking cleans 3 times with drinking water earlier, and reuse water for injection cleans 3 times.After the meat grinder rubbing, add cold water for injection 40L, with colloid mill homogenate 2min.Homogenate put-30 ℃ freezing 24 hours, take out 30 ℃ of water-baths next day and melt, 5 times repeatedly, be heated to 85 ℃ rapidly after melting for the 5th time and kept 5 minutes, speed is chilled to room temperature again.Centrifugal 20 minutes of 3000rpm gets supernatant liquid filtering, the 10000 daltonian ultrafilter ultrafiltration of filtrate reuse molecular cut off, and aseptic collection permeate is thymus peptide solution.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 9.6mg/ml, Thymosin alpha
1Account for 2.6% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
The piglets thymus 100Kg that gets the quick-freezing preservation puts in the special-purpose tank and melts, and extracts fat and adventitia, gets net weight 88Kg.The thymus of plucking cleans 3 times with drinking water earlier, and reuse water for injection cleans 3 times.After the meat grinder rubbing, add cold water for injection 88L, with colloid mill homogenate 2min.Homogenate put-30 ℃ freezing 48 hours, take out back 20 ℃ of water-baths and melt, be heated to 80 ℃ rapidly and kept 10 minutes, speed is chilled to room temperature.Centrifugal 20 minutes of 3000rpm gets supernatant liquid filtering, the 10000 daltonian ultrafilter ultrafiltration of filtrate reuse molecular cut off, and aseptic collection permeate is thymus peptide solution.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 6.1mg/ml, Thymosin alpha
1Account for 1.6% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
The piglets thymus 100Kg that gets the quick-freezing preservation puts in the special-purpose tank and melts, and extracts fat and adventitia, gets net weight 85Kg.The thymus of plucking cleans 3 times with drinking water earlier, and reuse water for injection cleans 3 times.After the meat grinder rubbing, add cold water for injection 60L, with colloid mill homogenate 2min.Homogenate put-30 ℃ freezing 24 hours, take out 25 ℃ of water-baths next day and melt, 3 times repeatedly, be heated to 85 ℃ rapidly after melting for the third time and kept 5 minutes, speed is chilled to room temperature again.Centrifugal 20 minutes of 3000rpm gets supernatant liquid filtering, the 10000 daltonian ultrafilter ultrafiltration of filtrate reuse molecular cut off, and aseptic collection permeate is thymus peptide solution.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 7.9mg/ml, Thymosin alpha
1Account for 2.2% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
The piglets thymus 100Kg that gets the quick-freezing preservation puts in the special-purpose tank and melts, and extracts fat and adventitia, gets net weight 90Kg.The thymus of plucking cleans 3 times with drinking water earlier, and reuse water for injection cleans 3 times.After the meat grinder rubbing, add cold water for injection 45L, with colloid mill homogenate 2min.Homogenate put-30 ℃ freezing 24 hours, take out 25 ℃ of water-baths next day and melt, 5 times repeatedly, be heated to 90 ℃ rapidly after melting for the 5th time and kept 3 minutes, speed is chilled to room temperature again.Centrifugal 20 minutes of 3000rpm gets supernatant liquid filtering, the 10000 daltonian ultrafilter ultrafiltration of filtrate reuse molecular cut off, and aseptic collection permeate is thymus peptide solution.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 9.8mg/ml, Thymosin alpha
1Account for 2.7% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
Fig. 3 is a suspension crystallization freeze concentration equipment sketch map.
Get the calf thymus peptide solution 100L that embodiment 1 obtains, by the scraper-type heat exchanger, thymus peptide solution is cooled to-1 ℃～1 ℃, makes the fine ice crystal of its generating portion, sends into the recrystallize jar with the 0.5L/min flow velocity.Because Ostwald's effect, little ice crystal melts, and big ice crystal is grown up, and this step needs 2 hours approximately.The thymus peptide solution that contains ice crystal is removed ice crystal by cleaning tower, ices melting liquid flushing ice crystal so that reclaim the concentrated solution that ice crystal surface adheres to part simultaneously, and flushing liquor is back to feed end and concentrates.Collect concentrated solution in clean container, obtain high concentration thymus peptide solution 10.5L.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 51.3mg/ml, Thymosin alpha
1Account for 1.6% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
Get the calf thymus peptide solution 100L that embodiment 2 obtains, by the scraper-type heat exchanger, thymus peptide solution is cooled to-2 ℃～2 ℃, makes the fine ice crystal of its generating portion send into the recrystallize jar with the 0.5L/min flow velocity.Because Ostwald's effect, little ice crystal melts, and big ice crystal is grown up, and this step needs 3 hours approximately.The thymus peptide solution that contains ice crystal is removed ice crystal by cleaning tower, ices melting liquid flushing ice crystal so that reclaim the concentrated solution that ice crystal surface adheres to part simultaneously, and flushing liquor is back to feed end and concentrates.Collect concentrated solution in clean container, obtain high concentration thymus peptide solution 12.8L.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 52.7mg/ml, Thymosin alpha
1Account for 2.1% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
Get the calf thymus peptide solution 100L that embodiment 3 obtains, by the scraper-type heat exchanger, thymus peptide solution is cooled to-4 ℃～4 ℃, makes the fine ice crystal of its generating portion send into the recrystallize jar with the 0.5L/min flow velocity.Because Ostwald's effect, little ice crystal melts, and big ice crystal is grown up, and this step needs 4 hours approximately.The thymus peptide solution that contains ice crystal is removed ice crystal by cleaning tower, ices melting liquid flushing ice crystal so that reclaim the concentrated solution that ice crystal surface adheres to part simultaneously, and flushing liquor is back to feed end and concentrates.Collect concentrated solution in clean container, obtain high concentration thymus peptide solution 16.1L.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 53.0mg/ml, Thymosin alpha
1Account for 2.6% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
Get the piglets thymus peptide solution 100L that embodiment 4 obtains, by the scraper-type heat exchanger, thymus peptide solution is cooled to-1 ℃～1 ℃, makes the fine ice crystal of its generating portion send into the recrystallize jar with the 0.5L/min flow velocity.Because Ostwald's effect, little ice crystal melts, and big ice crystal is grown up, and this step needs 3 hours approximately.The thymus peptide solution that contains ice crystal is removed ice crystal by cleaning tower, ices melting liquid flushing ice crystal so that reclaim the concentrated solution that ice crystal surface adheres to part simultaneously, and flushing liquor is back to feed end and concentrates.Collect concentrated solution in clean container, obtain high concentration thymus peptide solution 10.2L.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 54.5mg/ml, Thymosin alpha
1Account for 1.7% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
Get the piglets thymus peptide solution 100L that embodiment 5 obtains, by the scraper-type heat exchanger, thymus peptide solution is cooled to-2 ℃～2 ℃, makes the fine ice crystal of its generating portion send into the recrystallize jar with the 0.5L/min flow velocity.Because Ostwald's effect, little ice crystal melts, and big ice crystal is grown up, and this step needs 4 hours approximately.The thymus peptide solution that contains ice crystal is removed ice crystal by cleaning tower, ices melting liquid flushing ice crystal so that reclaim the concentrated solution that ice crystal surface adheres to part simultaneously, and flushing liquor is back to feed end and concentrates.Collect concentrated solution in clean container, obtain high concentration thymus peptide solution 13.2L.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 55.6mg/ml, Thymosin alpha
1Account for 2.2% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
Get the calf thymus peptide solution 100L that embodiment 6 obtains, by the scraper-type heat exchanger, thymus peptide solution is cooled to-4 ℃～4 ℃, makes the fine ice crystal of its generating portion send into the recrystallize jar with the 0.5L/min flow velocity.Because Ostwald's effect, little ice crystal melts, and big ice crystal is grown up, and this step needs 4 hours approximately.The thymus peptide solution that contains ice crystal is removed ice crystal by cleaning tower, ices melting liquid flushing ice crystal so that reclaim the concentrated solution that ice crystal surface adheres to part simultaneously, and flushing liquor is back to feed end and concentrates.Collect concentrated solution in clean container, obtain high concentration thymus peptide solution 15.7L.
Press thymus peptide solution national drug standards WS
1-XG-042-2000-2003 detects, and content is 57.3mg/ml, Thymosin alpha
1Account for 2.6% (standard-required must not be lower than 1.0%), molecular weight does not detect (standard code must not cross 5.0%) greater than 10000 daltonian high molecular weight materials, and character, discriminating, pH and protein inspection, activity are all up to specification.
At first, get high concentration calf thymus peptide solution 12～15L that embodiment 7～9 obtains respectively, carry out depyrogenation with 10000 daltonian ultrafilters and handle, collect ultrafiltrate in aseptic apyrogeneity container, measure through intermediate, content is 52.0mg/ml; Then, in the local laminar flow clean area, accurately measure above-mentioned intermediate 9.7L in liquid dispensing container by technology preparation, add aseptic pyrogen-free dextran solution as excipient, add the injection water again to 20.0L, fully mixing, aseptic filtration, in the 5ml control antibiotic bottle of packing into filtrate branch, every loading amount 2.0ml, sign indicating number is gone on the freezer dryer internal partition; Then, carry out lyophilizing, dried frozen aquatic products again through roll cover finished product; At last, after leak detection, lamp inspection and product inspection are qualified, pack again, go into warehouse for finished product and deposit.
At first, get high concentration piglets thymus peptide solution 11～14L that embodiment 10～12 obtains, carry out depyrogenation with 10000 daltonian ultrafilters and handle, collect ultrafiltrate in aseptic apyrogeneity container, measure through intermediate, content is 55.2mg/ml; Then, in the local laminar flow clean area, accurately measure above-mentioned intermediate 9.1L in liquid dispensing container by technology preparation, add aseptic pyrogen-free dextran solution as excipient, add the injection water again to 20.0L, fully mixing, aseptic filtration, in the 5ml control antibiotic bottle of packing into filtrate branch, every loading amount 2.0ml, sign indicating number is gone on the freezer dryer internal partition; Then, carry out lyophilizing, dried frozen aquatic products again through roll cover finished product; At last, after leak detection, lamp inspection and product inspection are qualified, pack again, go into warehouse for finished product and deposit.
At first, get high concentration calf thymus peptide solution 22～25L that embodiment 7～9 obtains, carry out depyrogenation with 10000 daltonian ultrafilters and handle, collect ultrafiltrate in aseptic apyrogeneity container, measure through intermediate, content is 52mg/ml; Then, in the local laminar flow clean area, accurately measure above-mentioned intermediate 19.3L in liquid dispensing container by technology preparation, add aseptic pyrogen-free mannitol solution as excipient, add the injection water again to 25.0L, fully mixing, aseptic filtration, in the 5ml control antibiotic bottle of packing into filtrate branch, every loading amount 2.5ml, sign indicating number is gone on the freezer dryer internal partition; Then, carry out lyophilizing, dried frozen aquatic products again through roll cover finished product; At last, after leak detection, lamp inspection and product inspection are qualified, pack again, go into warehouse for finished product and deposit.
At first, get high concentration piglets thymus peptide solution 21～24L that embodiment 10～12 obtains, carry out depyrogenation with 10000 daltonian ultrafilters and handle, collect ultrafiltrate in aseptic apyrogeneity container, measure through intermediate, content is 55.2mg/ml; Then, in the local laminar flow clean area, accurately measure above-mentioned intermediate 18.2L in liquid dispensing container by technology preparation, add aseptic pyrogen-free mannitol solution as excipient, add the injection water again to 25.0L, fully mixing, aseptic filtration, in the 5ml control antibiotic bottle of packing into filtrate branch, every loading amount 2.5ml, sign indicating number is gone on the freezer dryer internal partition; Then, carry out lyophilizing, dried frozen aquatic products again through roll cover finished product; At last, after leak detection, lamp inspection and product inspection are qualified, pack again, go into warehouse for finished product and deposit.
Claims (9)
1. the preparation method of a high concentration thymus peptide solution is characterized in that, may further comprise the steps:
1) from the thymus material below preparation molecular weight 10000 dalton, thymosin content is the low concentration thymus peptide solution of 5～10mg/ml;
2) concentrating of low concentration thymus peptide solution: adopt Freeze Concentration Technology to prepare the high concentration thymus peptide solution that thymosin content is 50～60mg/ml the low concentration thymus peptide solution that obtains in the step 1).
2. preparation method as claimed in claim 1 is characterized in that described thymus comprises calf thymus or porcine thymus.
3. preparation method as claimed in claim 1 is characterized in that described step 1) comprises
1a) get thymus fresh or that quick-freezing is preserved, remove impurity and clean; The thymus of handling well is added water be prepared into homogenate; With being refrigerated to deep layer rapidly under ℃ temperature of homogenate-25～-35, in 20～30 ℃ water-bath, melt multigelation then;
1b) heating freeze thawing liquid is removed unsettled foreign protein, again through centrifugal, mistake leaching supernatant; The component of supernatant below ultrafiltration molecular cut off 10000 dalton obtains the thymus peptide solution of low concentration.
4. as claim 1 or 3 described preparation methoies, it is characterized in that described step 2) comprising:
2a) the low concentration thymus peptide solution that obtains in the step 1) is cooled to-5～5 ℃, makes the solution after the described cooling generate ice crystal;
2b) above-mentioned steps continues 2～4 hours, makes ice crystal grow into big ice crystal;
2c) remove ice crystal, reclaim solution, obtain the thymus peptide solution of high concentration.
5. a high concentration thymus peptide solution preparation is characterized in that it contains the thymosin that is made by claim 1 method, and the content of wherein said thymosin is 50～60mg/ml.
6. the described high concentration Thymopeptide of claim 5 is characterized in that it further comprises pharmaceutically acceptable carrier and/or solvent.
7. one kind big specification thymosin lyophilized formulations is characterized in that, it is to be formed as feedstock production by the thymosin that claim 1 method makes, and the thymus peptide content is 100mg in the unit formulation.
8. the described big specification thymosin lyophilized formulations of claim 7 is characterized in that it further comprises pharmaceutically acceptable excipient.
9. the described big specification thymosin lyophilized formulations of claim 7 is characterized in that described unit formulation is each preparation packing.
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| CN1840177B (en) * | 2006-01-11 | 2011-04-06 | 成都圣诺科技发展有限公司 | Injection liquid of thymic peptide alpha 1 and preparation method thereof |
| CN101921331A (en) * | 2010-08-17 | 2010-12-22 | 郑州后羿制药有限公司 | Method for preparing thymosin |
| CN105777891A (en) * | 2014-12-16 | 2016-07-20 | 康普药业股份有限公司 | Method for improving extraction efficiency of thymosin |
| CN104856965B (en) * | 2015-05-29 | 2018-07-20 | 湖南科伦制药有限公司 | A kind of freeze drying process of injection thymic peptide |
| CN106138099A (en) * | 2016-07-12 | 2016-11-23 | 浙江华尔成生物药业股份有限公司 | Pharmaceutical composition, pharmaceutical preparation and preparation method thereof, purposes |
| CN108125139A (en) * | 2017-12-11 | 2018-06-08 | 长春三真实业有限公司 | A kind of thymic extract frozen dried food that can improve the elderly's body's immunity |
| CN107897602B (en) * | 2018-01-05 | 2023-10-03 | 陈锦权 | Movable horizontal type freeze concentration equipment and freeze concentration method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0331934A1 (en) * | 1988-03-11 | 1989-09-13 | SCLAVO S.p.A. | New pharmaceutical compositions containing thymosin alpha 1 |
| CN1152052C (en) * | 2002-05-16 | 2004-06-02 | 西安迪赛生物药业有限责任公司 | Process for preparing thymic peptide |
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2004
- 2004-06-23 CN CNB2004100092480A patent/CN100402085C/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0331934A1 (en) * | 1988-03-11 | 1989-09-13 | SCLAVO S.p.A. | New pharmaceutical compositions containing thymosin alpha 1 |
| CN1152052C (en) * | 2002-05-16 | 2004-06-02 | 西安迪赛生物药业有限责任公司 | Process for preparing thymic peptide |
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| Title |
|---|
| 胸腺素的两种制备工艺对比. 王凤山.氨基酸杂志，第2期. 1993 |
| 胸腺素的两种制备工艺对比. 王凤山.氨基酸杂志，第2期. 1993 * |
| 胸腺肽溶液质量标准. 中国药品标准，第4卷第6期. 2003 |
| 胸腺肽溶液质量标准. 中国药品标准，第4卷第6期. 2003 * |
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| Publication number | Publication date |
|---|---|
| CN1712064A (en) | 2005-12-28 |
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Address after: 100176 No. 8 prosperous street, Beijing economic and Technological Development Zone Patentee after: Beijing Science Sun Pharmaceutical Co., Ltd. Address before: 100176 No. 8 prosperous street, Beijing economic and Technological Development Zone Patentee before: Beijing Saisheng Pharmaceutical Co., Ltd. |