CN103739701A - Technical solution for purification of menotrophin - Google Patents
Technical solution for purification of menotrophin Download PDFInfo
- Publication number
- CN103739701A CN103739701A CN201310643509.3A CN201310643509A CN103739701A CN 103739701 A CN103739701 A CN 103739701A CN 201310643509 A CN201310643509 A CN 201310643509A CN 103739701 A CN103739701 A CN 103739701A
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- CN
- China
- Prior art keywords
- centrifugal
- supernatant liquor
- supernatant
- resin
- menotropins
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- 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.)
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- 108010057021 Menotropins Proteins 0.000 title claims abstract description 16
- 238000000746 purification Methods 0.000 title claims abstract description 6
- CBUWTGCATVNMJE-UHFFFAOYSA-N 6,6-dimethylheptanal Chemical compound CC(C)(C)CCCCC=O CBUWTGCATVNMJE-UHFFFAOYSA-N 0.000 title abstract 5
- 239000006228 supernatant Substances 0.000 claims abstract description 29
- 238000003756 stirring Methods 0.000 claims abstract description 22
- 239000011347 resin Substances 0.000 claims abstract description 15
- 229920005989 resin Polymers 0.000 claims abstract description 15
- 238000001035 drying Methods 0.000 claims abstract description 8
- 239000000843 powder Substances 0.000 claims abstract description 6
- 238000001556 precipitation Methods 0.000 claims abstract description 5
- 238000005406 washing Methods 0.000 claims abstract 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 27
- 229960004756 ethanol Drugs 0.000 claims description 12
- 238000013016 damping Methods 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 6
- 239000005695 Ammonium acetate Substances 0.000 claims description 6
- 229940043376 ammonium acetate Drugs 0.000 claims description 6
- 235000019257 ammonium acetate Nutrition 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000010521 absorption reaction Methods 0.000 claims description 5
- 239000012043 crude product Substances 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 3
- 229960000583 acetic acid Drugs 0.000 claims description 3
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 3
- 230000018044 dehydration Effects 0.000 claims description 3
- 238000006297 dehydration reaction Methods 0.000 claims description 3
- 239000012362 glacial acetic acid Substances 0.000 claims description 3
- 230000003252 repetitive effect Effects 0.000 claims description 3
- 238000005201 scrubbing Methods 0.000 claims description 3
- 239000001632 sodium acetate Substances 0.000 claims description 3
- 229960004249 sodium acetate Drugs 0.000 claims description 3
- 235000017281 sodium acetate Nutrition 0.000 claims description 3
- 239000007974 sodium acetate buffer Substances 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 239000000243 solution Substances 0.000 abstract description 7
- 239000007853 buffer solution Substances 0.000 abstract 1
- 238000010828 elution Methods 0.000 abstract 1
- 238000000605 extraction Methods 0.000 abstract 1
- 239000012535 impurity Substances 0.000 abstract 1
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 238000001179 sorption measurement Methods 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 description 4
- 210000002700 urine Anatomy 0.000 description 4
- GNTDGMZSJNCJKK-UHFFFAOYSA-N divanadium pentaoxide Chemical compound O=[V](=O)O[V](=O)=O GNTDGMZSJNCJKK-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 206010001928 Amenorrhoea Diseases 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 206010062767 Hypophysitis Diseases 0.000 description 1
- 108010073521 Luteinizing Hormone Proteins 0.000 description 1
- 102000009151 Luteinizing Hormone Human genes 0.000 description 1
- 208000019255 Menstrual disease Diseases 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 102000011759 adducin Human genes 0.000 description 1
- 108010076723 adducin Proteins 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000003325 follicular Effects 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000003821 menstrual periods Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000036299 sexual function Effects 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Endocrinology (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Reproductive Health (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a technical solution for purification of menotrophin by adsorption, elution, precipitation, drying and other steps, the technical solution is characterized in that dissolving menotrophin coarse extraction powder in a buffer solution, adding resin to a supernatant, stirring, centrifuging, removing the supernatant, washing the resin, adding an eluant, stirring, centrifuging, precipitating, and drying to obtain Kyowa dry powder, and the technical solution has the advantages of removal of most of impurities in the menotrophin and purification of the menotrophin.
Description
Technical field
The invention discloses a kind of Menotropins crude product through must the coordinate technical scheme of dry powder of absorption, wash-out, precipitation, drying and other steps purification Menotropins
Background technology
Menotropins (HMG) is by the merocrine a kind of glycoprotein gonad-stimulating hormone of people's anterior lobe of hypophysis, is generally through extracting purifying, to make in the urine of women from climacteric or climacteric.Women's content of HMG between menstrual period fluctuates to some extent, but lower, only have women to enter in climacteric or more than ten years afterwards, in its urine, (60-100iu/L urine) peaks the content side of HMG, and can maintain in a stable level, HMG contains follicular stimulating hormone (FSH) and lutropin (LH), and its contained ratio is similar to 1:1.Clinically be mainly used in treating women because of functional infertility due to not ovulating; The male sex is because of the low caused spermacrasia of hormonal readiness or vigor deficiency.In addition, be also used for the treatment of diseases such as women's amenorrhoea, menoxenia, the imbalance of men and women's sexual function.
First nineteen twenty-eight Ascbheim has reported from climacteric women urine and has found active substance, though report is many afterwards, but real to the understanding of HMG and preparation, is the Donimi since the sixties in 20th century, and forms gradually prepared by more normalized technique.China begins one's study from 20 century 70s, formally enters clinical experimental stage the eighties, starts a large amount of production the nineties.
Summary of the invention
The present invention discloses a kind of Menotropins crude product through the technical scheme of absorption, wash-out, precipitation, drying and other steps purification Menotropins.
1. damping fluid compound method is: take a certain amount of sodium-acetate, add water stirring and dissolving, drip Glacial acetic acid and adjust pH;
2. elutriant compound method is: by 95% ethanol: water stirs in the ratio of 0.4-0.5:0.5-0.6 (L:L), is made into 40% ethanol, then ammonium acetate is dissolved in 40% ethanol, make every liter in containing ammonium acetate 90-120g;
3. absorption:
By Menotropins crude product: damping fluid is in the ratio of 1-2:100-150 (Kg:L) stirring and dissolving under cold condition, centrifuging and taking supernatant liquor, supernatant liquor after centrifugal is added to consonance resin agitating 2 hours by 1-1.5:10-20 (L:L) again, centrifugal abandoning supernatant, then uses resin sodium-acetate buffer repetitive scrubbing;
4. wash-out
The resin having washed is added to elutriant by 0.5-1.5:8-20 (L:L), stir 2 hours, centrifugal, obtain supernatant liquor, then add elutriant in 0.5-1.5:5-15 (L:L) ratio, stir, centrifugal, obtain supernatant liquor;
5. precipitate
Merge twice supernatant liquor and add 2-6 doubly to measure ethanol, under 8-12 ℃ of condition, stir 30 minutes standing over night;
6. dry
Siphon supernatant liquor, throw out is centrifugal abandons supernatant afterwards with dehydrated alcohol dehydration twice, the throw out vacuum drying oven inner drying dry product of must coordinating.
Embodiment
Embodiment 1
1. the preparation of solution:
The preparation of 1.1 damping fluids: take a certain amount of sodium-acetate, add appropriate purified water stirring and dissolving, drip Glacial acetic acid and adjust PH.
The preparation of 1.2 elutriants: 0.421 part of 95% ethanol adds 0.579 part of water, stirs, and is made into 40% ethanol, then ammonium acetate is dissolved in 40% ethanol, making ammonium acetate content in every liter of solution is 100g.
2. absorption
2.1 first will slightly promote powder doubly measures stirring and dissolving under low temperature 8-12 ℃ condition, centrifuging and taking supernatant liquor by damping fluid (1:100).
2.2 supernatant liquors add 10 times of amount consonance resin agitatings of product 2 hours again, centrifugal abandoning supernatant.Resin sodium-acetate buffer repetitive scrubbing, is less than till 0.25 to 280nmOD value.
3. wash-out
8 times of amounts of the resin having adsorbed are added to elutriant, stir 2 hours, centrifugal, obtain supernatant.Resin adds 6 times of amount elutriants again, stirs 2 hours, centrifugal, obtains supernatant, discards resin.
4. precipitate
Merge supernatant twice, proceed in stainless steel cask, while stirring, add 95% ethanol of 4 times of amounts, under low temperature 8-12 ℃ condition, stir 30 minutes static spending the night.
5. dry
Siphon 5.1 next day abandoning supernatant, throw out is centrifugal.
Appropriate dehydrated alcohol dehydration 2 times for 5.2 throw outs, centrifugal, abandon supernatant.
5.3 last throw outs are equipped with the vacuum drying oven inner drying that Vanadium Pentoxide in FLAKES is done water-retaining agent, the dry product of must coordinating after being dried.
The above, be only preferred embodiment of the present invention, is not the present invention to be done to the restriction of other form, and any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the equivalent embodiment of equivalent variations.But every any simple modification, equivalent variations and remodeling that does not depart from technical solution of the present invention content, above embodiment is done according to technical spirit of the present invention, still belongs to the protection domain of technical solution of the present invention.
Claims (8)
1. the present invention discloses a kind of Menotropins crude product through the technical scheme of absorption, wash-out, precipitation, drying and other steps purification Menotropins, it is characterized in that Menotropins is slightly promoted to powder to be dissolved in damping fluid, again supernatant liquor is added to resin agitating, centrifugal abandoning supernatant, after washing resin, add again elutriant, stir, centrifugal, obtain supernatant, precipitation, the dry consonance dry powder that to obtain.
2. according to damping fluid compound method described in claims 1, be: take a certain amount of sodium-acetate, add water stirring and dissolving, drip Glacial acetic acid and adjust pH.
3. according to elutriant compound method described in claims 1, be: by 95% ethanol: water is in (0.4-0.5): (0.5-0.6) ratio of (L:L) stirs, be made into 40% ethanol, again ammonium acetate is dissolved in 40% ethanol, make every liter in containing ammonium acetate 90-120g.
According to described in claims 1 by Menotropins crude product: damping fluid is in (1-2): (100-150) ratio of (Kg:L) stirring and dissolving under cold condition, centrifuging and taking supernatant liquor.
5. according to the supernatant liquor after centrifugal described in claims 1, press again (1-1.5): (10-20) (L:L) adds consonance resin agitating 2 hours, and centrifugal abandoning supernatant, then uses resin sodium-acetate buffer repetitive scrubbing.
6. according to described in claims 1, the resin having washed being pressed to (0.5-1.5): (8-20) (L:L) adds elutriant, stir 2 hours, centrifugal, obtain supernatant liquor, again in (0.5-1.5): (5-15) (L:L) ratio adds elutriant, stir, centrifugal, obtain supernatant liquor.
7. merge according to claim 1 twice supernatant liquor and add 2-6 doubly to measure ethanol, under 8-12 ℃ of condition, stir 30 minutes standing over night.
8. siphon supernatant liquor according to claim 1, throw out is centrifugal abandons supernatant afterwards with dehydrated alcohol dehydration twice, the throw out vacuum drying oven inner drying dry product of must coordinating.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310643509.3A CN103739701A (en) | 2013-11-30 | 2013-11-30 | Technical solution for purification of menotrophin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310643509.3A CN103739701A (en) | 2013-11-30 | 2013-11-30 | Technical solution for purification of menotrophin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103739701A true CN103739701A (en) | 2014-04-23 |
Family
ID=50496785
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310643509.3A Pending CN103739701A (en) | 2013-11-30 | 2013-11-30 | Technical solution for purification of menotrophin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103739701A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105399813A (en) * | 2015-11-26 | 2016-03-16 | 青岛康原药业有限公司 | Method for extracting and purifying high-valence HMG (human menopausal gonadotropin) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1958603A (en) * | 2005-11-04 | 2007-05-09 | 上海天伟生物制药有限公司 | Method for purifying human chorionic gonadotropin |
| CN101792482A (en) * | 2010-03-12 | 2010-08-04 | 丽珠医药集团股份有限公司 | Method for purifying gonadotropins |
| CN101792481A (en) * | 2010-03-12 | 2010-08-04 | 丽珠医药集团股份有限公司 | Purification method of human chorionic gonadotropin (HCG) |
-
2013
- 2013-11-30 CN CN201310643509.3A patent/CN103739701A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1958603A (en) * | 2005-11-04 | 2007-05-09 | 上海天伟生物制药有限公司 | Method for purifying human chorionic gonadotropin |
| CN101792482A (en) * | 2010-03-12 | 2010-08-04 | 丽珠医药集团股份有限公司 | Method for purifying gonadotropins |
| CN101792481A (en) * | 2010-03-12 | 2010-08-04 | 丽珠医药集团股份有限公司 | Purification method of human chorionic gonadotropin (HCG) |
Non-Patent Citations (1)
| Title |
|---|
| 邹国林等: "尿促性腺激素", 《生化药物杂志》, no. 3, 1 October 1991 (1991-10-01) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105399813A (en) * | 2015-11-26 | 2016-03-16 | 青岛康原药业有限公司 | Method for extracting and purifying high-valence HMG (human menopausal gonadotropin) |
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| PB01 | Publication | ||
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| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
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Application publication date: 20140423 |