CN109942697A - A kind of rework preocess of corticotropin(ACTH) - Google Patents
A kind of rework preocess of corticotropin(ACTH) Download PDFInfo
- Publication number
- CN109942697A CN109942697A CN201910243773.5A CN201910243773A CN109942697A CN 109942697 A CN109942697 A CN 109942697A CN 201910243773 A CN201910243773 A CN 201910243773A CN 109942697 A CN109942697 A CN 109942697A
- Authority
- CN
- China
- Prior art keywords
- acth
- corticotropin
- finished product
- over again
- ice
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a kind of rework preocess of corticotropin(ACTH), relate in particular to following steps: the finished product done over again will be needed to be placed in stainless steel barrel, purified water, stirring and dissolving is added；Dissolved finished product is refrigerated to icing；After freezing completely, then it is made to change ice；After completion ice, filtered on buchner funnel is used；Organic solution is added into filtered filtrate to be precipitated, after the completion of precipitating, removes supernatant, sediment is dehydrated with acetone and is washed；Dewatered drying precipitate is handled, corticotropin(ACTH) finished product is obtained.Corticotropin(ACTH) rework preocess of the invention is simple, and at low cost, process is mild, can effectively solve the underproof problem of product clarity, and product is made to regain economic value, creates income, retrieves a loss and avoid to waste；The corticotropin(ACTH) produced using technical solution of the present invention, compared to before doing over again, the corticotropin(ACTH) potency after doing over again improves about 0.24U/mg.
Description
Technical field
The present invention relates to field of biotechnology more particularly to a kind of rework preocess of corticotropin(ACTH).
Background technique
Corticotropin(ACTH) (Corticotrophin, ACTH) is a kind of protein and peptide hormone, promotes glucocorticoid secretion, dimension
Hold the normal function of adrenocorticotropic.Natural ACTH is a straight-chain polypeptide, is made of 39 amino acid, molecular weight 4500
~4600 dalton (Da), it is soluble easily in water, it can also be dissolved in 60~70% acetone and ethyl alcohol.In dry and acid solution compared with
Stablize, under 100 DEG C of heating, ACTH vigor is unaffected；And the easy in inactivation in the solution of alkalinity.24 peptides of ACTH, i.e., 1
~24 segments (ACTH 1~24) have all activity, and 15 amino acid residues later be immunized with kind it is related, no
The effect for participating in isoacceptor, only maintains the stability of entire polypeptide structure.Therefore artificial synthesized ACTH 1~24 and natural A CTH
Act on immune response that is identical, and not causing body.
It both includes the key enzyme 3- hydroxyl -3- first-penta two for promoting cholesterol biosynthesis that ACTH, which promotes the mechanism of cortin synthesis,
Acyl coenzyme A (HMG-CoA) reductase activity, increases cholesterol biosynthesis；It also include increasing cholesterol side chain oxicracking enzyme system
Activity make cholesterol be transformed into pregnenolone increase, so that corticosteroid be promoted to synthesize.
As one very important product of our company, while the enterprise as national unique production ACTH, ACTH product
Yield and company's benefit moment are linked up with.The index of correlation of usual ACTH quality testing mainly has 5, and respectively character, solution is clear
Clear degree, hypertensive substance, undue toxicity and potency.In testing result, it often and only will appear the underproof situation of clarity,
And once occur that clarity of solution is unqualified to be will lead to entire block and directly scrap, other than income is substantially damaged, also waste
A large amount of raw material and manpower and material resources.Therefore, for the underproof product of these clarity how to retrieve certain loss at
It is eager to solve the problems, such as.
Summary of the invention
In view of the foregoing deficiencies of prior art, the purpose of the present invention is to provide a kind of works of doing over again of corticotropin(ACTH)
Skill faces and scraps because product clarity is unqualified in production for solving, causes asking for the wastes such as human and material resources and financial resources
Topic, retrieves a loss as far as possible.
In order to achieve the above objects and other related objects, on the one hand, the present invention provides a kind of works of doing over again of corticotropin(ACTH)
Skill, chilled to change ice processing again after the finished product dissolution that needs are done over again, filtering, precipitating and drying obtain corticotropin(ACTH) finished product.
Specifically, which includes the following steps:
1) it dissolves: the finished product done over again will be needed to be placed in stainless steel barrel, purified water is added, stirring is extremely dissolved；
2) it freezes: finished product dissolved in step 1) is refrigerated to icing；
3) change ice: after freezing completely in step 2), then it being made to change ice；
4) it filters: after completion ice in step 3), clarification plate and two layers of filter paper filtering is put into Buchner funnel；
5) it precipitates: organic solution being added into filtrate filtered in step 4) and is precipitated, after the completion of precipitating, goes
Clearly, sediment is dehydrated with acetone and is washed；
Wherein, acetone is preferably analyzed pure, and main purpose is in order to which finished product is further dried；
6) dry: by drying precipitate dewatered in step 5) processing, to obtain corticotropin(ACTH) finished product.
Preferably, in step 1), the mass ratio of the pure water and the finished product for needing to do over again is 15~25:1, further
Preferably, the mass ratio of the pure water and the finished product for needing to do over again is 20:1
Preferably, in step 2), dissolved finished product is refrigerated to icing method particularly includes: be placed in stainless steel barrel interior
Wall is connected in the enamel pot of ethylene glycol, is refrigerated to icing, the gap processing industry third between stainless steel barrel and enamel pot
Ketone is as medium, and while to guarantee that solution is cooled uniform in bucket, stainless steel barrel, which will not be frozen, to be difficult to take out in pot.
Preferably, the ethylene glycol ice bath is closed after solution freezes completely, makes its naturalization ice.
Preferably, in step 5), the volume ratio of the organic solution and the filtrate is 8~12:1, it is further preferred that
The organic solution and the volume ratio of the filtrate are 10:1.
It is further preferred that the acetone that the organic solution is 0~-10 DEG C, wherein preferably, the acetone is industry
Acetone can reduce rework cost using industrial acetone.
Preferably, the drying process method particularly includes: be dried in vacuo dewatered sediment with phosphorus pentoxide
40~48 hours.The purpose that phosphorus pentoxide is added is the vapor in absorption dehydration case.
On the other hand, the present invention also provides a kind of corticotropin(ACTH) obtained by above-mentioned rework preocess.
As described above, a kind of rework preocess of corticotropin(ACTH) provided by the invention, has the advantages that
(1) corticotropin(ACTH) rework preocess of the invention is simple, and at low cost, process is mild；
(2) compared to before doing over again, the corticotropin(ACTH) potency after doing over again improves about 0.24U/mg；
(3) being done over again using corticotropin(ACTH) rework preocess of the invention can effectively solve the underproof problem of product clarity,
So that product is regained economic value, creates income, retrieve a loss and avoid to waste.
Detailed description of the invention
Fig. 1 be the embodiment of the present invention 2 in ATCH do over again front and back potency comparison line chart；
Fig. 2 is that ATCH does over again front and back yield comparison line chart in the embodiment of the present invention 2；
Fig. 3 be the embodiment of the present invention 2 in ATCH do over again front and back yield comparison line chart.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
Experimental method described in following embodiments is unless otherwise specified conventional method, the reagent and material for example without
Specified otherwise commercially obtains.
The quality standard of ACTH finished product is specifically shown in the following table 1:
1 ACTH final product quality standard of table
Embodiment 1:
There are 10 batches to do over again because clarity of solution is unqualified in 24 batches of ACTH products of the production of our company in 2018.First will
Chilled to change ice processing again after the finished product dissolution for needing to do over again, filtering, precipitating and drying obtain corticotropin(ACTH) finished product.Needs are returned
Chilled to change ice processing again after the finished product dissolution of work, filtering, precipitating and drying obtain corticotropin(ACTH) finished product.Specifically, the technique
Include the following steps:
1) it dissolves: the finished product done over again will be needed to be placed in stainless steel barrel, 20 times of purified water is then added, stir about 1 is small
When to ensure that it is completely dissolved；
2) it freezes: stainless steel barrel being placed in the enamel pot of the logical ethylene glycol of inner wall and be refrigerated to icing.And in stainless steel barrel
Gap processing industry acetone between enamel pot is as medium, while to guarantee that solution is cooled uniform in bucket, stainless steel
Bucket, which will not be frozen, to be difficult to take out in pot.
3) change ice: closing ethylene glycol ice bath after solution freezes completely, make its naturalization ice；
4) it filters: after changing ice, clarification plate is added in Buchner funnel and two layers of filter paper is filtered；
5) precipitate: the industrial acetone that filtrate measures 0~-10 DEG C with 10 times is precipitated, so that ACTH is separated from water；It has precipitated
Cheng Hou removes supernatant, and sediment is washed with acetone (analysis is pure) dehydration, to achieve the purpose that finished product is further dried；
6) dry: dewatered sediment phosphorus pentoxide being dried in vacuo 40~48 hours, corticotropin(ACTH) finished product is obtained.
The purpose that phosphorus pentoxide is added is the vapor in absorption dehydration case.
Embodiment 2:
Performance evaluation is carried out to 10 batches of finished products after doing over again according to the rework preocess in embodiment 1.In every quality standard
In, product characteristics, hypertensive substance and undue toxicity in internal control critical field, in the present embodiment only to the clarity of product,
Potency and relevant item are illustrated.
(1) ACTH do over again front and back clarity comparison
Clarity checks that the genealogy of law compares drug solution with regulation turbidity standard, is used to check the clarification journey of solution
Degree, it is specified that under " clarification " refer to that the clarity of drug solution is identical as solvent for use, or it is turbid no more than No. 0.5 turbidity standard
Degree.
1) preparation of No. 0.5 turbidity standard
It weighs in 105 DEG C of dry hydrazine sulfate 1.00g to constant weight, sets in 100mL measuring bottle, appropriate amount of water is added to make to dissolve, it is necessary
When can be in 40 DEG C of water-bath medium temperature heat of solution, and be diluted with water to scale, shake up, place 4~6 hours；It takes this solution and waits and hold
The 10% methenamine solution mixing of amount, shakes up, and standing is protected from light in 25 DEG C 24 hours to get Standard Reserving Solution.
Standard Reserving Solution 15.0mL is taken, sets in 1000mL measuring bottle, is diluted with water to scale, is shaken up to get turbidity standard original
Liquid.It takes in right amount, is placed in 1cm absorption cell, using ultraviolet-visible scene photometry, its extinction is measured under the wavelength of 550nm
Degree, absorbance should be in 0.12~0.15 range.
It takes 2.50mL turbidity standard stoste to mix with 97.50mL water, sufficiently shakes up to get No. 0.5 turbidity standard.
2) visual method checks clarity
No. 0.5 turbidity standard for being diluted with water to certain density drug solution and equivalent is distinguished at room temperature
The ratio for being placed in pairing is turbid with glass tube (15~16mm of internal diameter, flat, tool plug is made with colourless, transparent, neutral hard glass)
In, it prepares after five minutes, is vertically arranged in darkroom under umbrella canopy lamp, illumination 10001x is seen from horizontal direction in turbidity standard
It examines, compare clarity.Experimental result is as shown in table 2 below:
2 ATCH of table do over again front and back clarity comparison
| Batch | Before doing over again | After doing over again |
| 1 | It is unqualified | It is qualified |
| 2 | It is unqualified | It is qualified |
| 3 | It is unqualified | It is qualified |
| 4 | It is unqualified | It is qualified |
| 5 | It is unqualified | It is qualified |
| 6 | It is unqualified | It is qualified |
| 7 | It is unqualified | It is qualified |
| 8 | It is unqualified | It is qualified |
| 9 | It is unqualified | It is qualified |
| 10 | It is unqualified | It is qualified |
(2) ATCH do over again front and back potency, yield and yield comparison
ATCH does over again the potency of front and back, the correlation data of yield and yield respectively as shown in the following table 3,4 and 5:
3 ATCH of table do over again front and back potency (U/mg) comparison
| Batch | Before doing over again | After doing over again |
| 1 | 1.39 | 1.53 |
| 2 | 1.44 | 1.54 |
| 3 | 1.27 | 1.55 |
| 4 | 1.36 | 1.53 |
| 5 | 1.26 | 1.76 |
| 6 | 1.43 | 1.69 |
| 7 | It does not detect | 1.48 |
| 8 | It does not detect | 1.70 |
| 9 | It does not detect | 1.48 |
| 10 | It does not detect | 2.08 |
* due to before the doing over again of number 7,8,9,10 potency do not detect so not showing in the graph
4 ATCH of table do over again front and back yield (g) comparison
| Batch | Before doing over again | After doing over again |
| 1 | 272 | 199 |
| 2 | 301 | 215 |
| 3 | 319 | 237 |
| 4 | 336 | 252 |
| 5 | 369 | 265 |
| 6 | 278 | 197 |
| 7 | 221 | 140 |
| 8 | 228 | 159 |
| 9 | 155 | 107 |
| 10 | 288 | 207 |
5 ATCH of table do over again front and back yield (ten thousand U/kg) comparison
| Batch | Before doing over again | After doing over again |
| 1 | 11.5 | 9.2 |
| 2 | 13.1 | 10.3 |
| 3 | 12.3 | 11.1 |
| 4 | 13.8 | 11.7 |
| 5 | 14.1 | 14.1 |
| 6 | 12.0 | 10.1 |
| 7 | It does not detect | 6.3 |
| 8 | It does not detect | 8.2 |
| 9 | It does not detect | 4.8 |
| 10 | It does not detect | 13.0 |
From table 2 it can be seen that product clarity after being done over again using corticotropin(ACTH) rework preocess of the invention all from
It is unqualified to become qualified；As can be seen that potency of the ACTH product after doing over again integrally is higher than and does over again from the data of table 3 and Fig. 1
Preceding potency, product potency averagely improve about 0.24U/mg；As can be seen that ACTH product is being done over again from the data of table 4 and Fig. 2
Yield afterwards is integrally lower than the yield before doing over again, and yield averagely declines about 85g；It can be seen that from the data of table 5 and Fig. 3
Yield of the ACTH product in front and back of doing over again is close, although most of is all that yield before doing over again is higher, the yield that number is 5 batches
It is almost equal before and after doing over again.
According to an injection 27.5U, 37 yuan/of tax-free price, average every batch of yield 227.5g, average every batch of potency
1.6U/mg is calculated, and the ACTH product after average every a batch is done over again can obtain about 4,897,450,000 yuan of income, is created not small
Economic value, retrieved many loss and waste.
Specific embodiments of the present invention are described in detail above, it should be understood that those skilled in the art are without wound
The property made labour, which according to the present invention can conceive, makes many modifications and variations.Therefore, all technician in the art
Pass through the available technology of logical analysis, reasoning, or a limited experiment on the basis of existing technology under this invention's idea
Scheme, all should be within the scope of protection determined by the claims.
Claims (9)
1. a kind of rework preocess of corticotropin(ACTH), which is characterized in that chilled to change at ice again after the finished product dissolution for needing to do over again
Reason, filtering, precipitating and drying, obtains corticotropin(ACTH) finished product.
2. the rework preocess of corticotropin(ACTH) according to claim 1, which is characterized in that the technique includes the following steps:
1) it dissolves: the finished product done over again will be needed to be placed in stainless steel barrel, purified water, stirring and dissolving is added；
2) it freezes: finished product dissolved in step 1) is refrigerated to icing；
3) change ice: after freezing completely in step 2), then it being made to change ice；
4) it filters: after completion ice in step 3), using filtered on buchner funnel；
5) it precipitates: organic solution being added into filtrate filtered in step 4) and is precipitated, after the completion of precipitating, removes supernatant, sinks
Starch is dehydrated with acetone and is washed；
6) dry: by drying precipitate dewatered in step 5) processing, to obtain corticotropin(ACTH) finished product.
3. the rework preocess of corticotropin(ACTH) according to claim 2, which is characterized in that in step 1), the pure water and institute
It states and needs the mass ratio for the finished product done over again for 15~25:1.
4. the rework preocess of corticotropin(ACTH) according to claim 2, which is characterized in that in step 2), by it is dissolved at
Product are refrigerated to icing method particularly includes: stainless steel barrel is placed in inner wall and is connected in the enamel pot of ethylene glycol, icing is refrigerated to.
5. the rework preocess of corticotropin(ACTH) according to claim 4, which is characterized in that close institute after solution freezes completely
Ethylene glycol ice bath is stated, its naturalization ice is made.
6. the rework preocess of corticotropin(ACTH) according to claim 2, which is characterized in that in step 5), the organic solution
Volume ratio with the filtrate is 8~12:1.
7. the rework preocess of corticotropin(ACTH) according to claim 6, which is characterized in that the organic solution is 0~-10 DEG C
Acetone.
8. the rework preocess of corticotropin(ACTH) according to claim 2, which is characterized in that in step 6), the drying process
Method particularly includes: dewatered sediment phosphorus pentoxide is dried in vacuo 40~48 hours.
9. a kind of corticotropin(ACTH) that the rework preocess by any corticotropin(ACTH) of claim 1~8 obtains.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910243773.5A CN109942697A (en) | 2019-03-28 | 2019-03-28 | A kind of rework preocess of corticotropin(ACTH) |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910243773.5A CN109942697A (en) | 2019-03-28 | 2019-03-28 | A kind of rework preocess of corticotropin(ACTH) |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109942697A true CN109942697A (en) | 2019-06-28 |
Family
ID=67012233
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910243773.5A Pending CN109942697A (en) | 2019-03-28 | 2019-03-28 | A kind of rework preocess of corticotropin(ACTH) |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109942697A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2906667A (en) * | 1956-08-21 | 1959-09-29 | Canada Packers Ltd | Preparation and purification of corticotrophin |
| CN103694341A (en) * | 2013-11-23 | 2014-04-02 | 青岛康原药业有限公司 | Preparation method of corticotropin |
| CN103724427A (en) * | 2013-11-29 | 2014-04-16 | 青岛康原药业有限公司 | Method for refining corticotropin |
| CN108948180A (en) * | 2018-06-26 | 2018-12-07 | 滨州医学院 | Corticotropin preparation method |
-
2019
- 2019-03-28 CN CN201910243773.5A patent/CN109942697A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2906667A (en) * | 1956-08-21 | 1959-09-29 | Canada Packers Ltd | Preparation and purification of corticotrophin |
| CN103694341A (en) * | 2013-11-23 | 2014-04-02 | 青岛康原药业有限公司 | Preparation method of corticotropin |
| CN103724427A (en) * | 2013-11-29 | 2014-04-16 | 青岛康原药业有限公司 | Method for refining corticotropin |
| CN108948180A (en) * | 2018-06-26 | 2018-12-07 | 滨州医学院 | Corticotropin preparation method |
Non-Patent Citations (6)
| Title |
|---|
| HEWETT CL等: "Extraction of ACTH from human pituitaries", 《NATURE》 * |
| 中国法制出版社: "《中华人民共和国卫生健康法律法规全书》", 31 January 2019 * |
| 国家基本药物领导小组: "《国家基本药物 西药》", 31 August 1999 * |
| 张炤化: "用盐酸丙酮法提取促皮质素（ACTH）的研究", 《化学世界》 * |
| 沈力: "《药事管理与法规》", 31 January 2017 * |
| 金文进: "《药物分析》", 31 January 2015 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xu et al. | An in situ hydrogel based on carboxymethyl chitosan and sodium alginate dialdehyde for corneal wound healing after alkali burn | |
| Tsuji et al. | Direct transformation from amorphous to crystalline calcium phosphate facilitated by motif-programmed artificial proteins | |
| Liu et al. | The role of matrix proteins in the control of nacreous layer deposition during pearl formation | |
| Bourgade et al. | Impact of the length of time between diagnosis and surgical removal of urologic neoplasms on survival | |
| Zand et al. | pH-dependent conformational changes of concanavalin A | |
| Kweon et al. | Structural and conformational changes of regenerated Antheraea pernyi silk fibroin films treated with methanol solution | |
| CN104172200A (en) | Method for increasing content of soluble dietary fibers in apple pomace | |
| Clarke et al. | The preparation of plant nucleic acids | |
| CN102585038A (en) | Islamic enoxaparin sodium and method for producing and purifying same | |
| Castillo et al. | Low levels of von Willebrand markers associate with high serum IgM levels and improve with response to therapy, in patients with Waldenström macroglobulinaemia. | |
| CN109942697A (en) | A kind of rework preocess of corticotropin(ACTH) | |
| Tooke et al. | Kinetics of the self‐aggregation and film formation of poly‐L‐proline at high temperatures explored by circular dichroism spectroscopy | |
| He et al. | Low pressure‐induced secondary structure transitions of regenerated silk fibroin in its wet film studied by time‐resolved infrared spectroscopy | |
| CN104568804B (en) | Basin total nitrogen total phosphorus detection method | |
| CN117209624A (en) | Chondroitin sulfate extraction process | |
| CN106188341B (en) | Heparin sodium leftover bits and pieces prepares high-purity sulfuric acid dermatan technique | |
| Jyothi et al. | Studies on the properties of citrate derivatives of cassava (Manihot esculenta Crantz) starch synthesized by microwave technique | |
| Wu et al. | Methanol–water-dependent structural changes of regenerated silk fibroin probed using terahertz spectroscopy | |
| CN110746659A (en) | Natural preservative film and preparation method thereof | |
| CN103755838A (en) | Preparation method of heparin sodium | |
| Rousselin et al. | Renal and vascular outcomes in patients with isolated antiphospholipid syndrome nephropathy | |
| Ginsberg et al. | Sensitive methods for the determination of ester sulfate in biological systems | |
| JPS59113001A (en) | Production of carrageenan | |
| Belton et al. | Traditional materials from new sources–conflicts in analytical methods for calcium carbonate | |
| CN110751401A (en) | Method for evaluating quality of encarsia formosa bee card |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |