CN104651439A - Enzymatic preparation process of 7-aminocephalosporanic acid - Google Patents
Enzymatic preparation process of 7-aminocephalosporanic acid Download PDFInfo
- Publication number
- CN104651439A CN104651439A CN201510127375.9A CN201510127375A CN104651439A CN 104651439 A CN104651439 A CN 104651439A CN 201510127375 A CN201510127375 A CN 201510127375A CN 104651439 A CN104651439 A CN 104651439A
- Authority
- CN
- China
- Prior art keywords
- amino
- cephalosporanic acid
- enzyme preparation
- preparation technique
- cephalosporin
- 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
- Cephalosporin Compounds (AREA)
Abstract
The invention discloses an enzymatic preparation process of 7-aminocephalosporanic acid, relating to the technical field of medicinal intermediate production. The process comprises the steps of carrying out pyrolysis on cephalosporin C; dezymotizing; carrying out ultrafiltration and the like. The content of the obtained 7-aminocephalosporanic acid is more than 99%, the color grade of the obtained 7-aminocephalosporanic acid is up to 2#, meanwhile, the indexes such as endotoxin, proteins and polymers are effectively improved, and the 7-aminocephalosporanic acid can be directly used for cephalosporin antibiotic production without purification; and the whole process is simple in operation, suitable for application to industrial production and capable of creating considerable economic and social benefits.
Description
Technical field
The invention belongs to medical art, relate to a kind of preparation method of medicine intermediate, be specifically related to a kind of enzyme preparation technique of 7-amino-cephalosporanic acid.
Background technology
7-amino-cephalosporanic acid, be called for short 7-ACA, molecular formula is C
10h
12n
2o
5s, molecular weight is 272.28, and chemical name is 3-acetyl-o-methyl-5-sulphur-7-amino-8-oxygen-1-azabicyclic oct-2-ene-2 carboxylic acid, and its structural formula is as follows:
7-amino-cephalosporanic acid is intermediate parent nucleus crucial in cephalosporins preparation, two active groups are had in its structure, the acetoxyl group of 3-position and the amino of 7-position, these two active groups connect different side chains, just form cephalosporin analog antibiotic of different nature, can be used for the production of cephalosporins, as cefotaxime (cefotaxime), rocephin (ceftriaxone), Kefzol (cefazolin), cephalofruxin (cefuroxime), cefoperazone (cefoperzone) etc.Over nearly 10 years, cephalosporins development is very surprising, constantly has new variety to go on the market, as Cefaclor, Cefuroxime sodium etc.The cephalosporins sum put into production at present is more than 50, and clinical conventional kind, also more than 30, has become the leading role in current international microbiotic market.
Thus, the status of 7-amino-cephalosporanic acid in cephalosporins is produced has some idea of, not only of crucial importance, and can not be substituted at present, so also just seem of crucial importance to the research and development of 7-amino-cephalosporanic acid production technique, no matter be Improving The Quality of Products, or reduce production cost, various improvement all by promoting the progress of pharmaceutical industries, for manufacturing enterprise creates objective economic benefit and social benefit.
At present, the 7-amino-cephalosporanic acid that cephalosporins uses in producing is all that the cephalosporin obtained by fermenting corn steep liquor obtains in amido linkage cracking, according to the difference of cleavage method, can be divided into chemical method and enzyme process.
Chemical method produces 7-ACA by cracking cephalosporin zinc salt or sodium salt, its reaction process needs to carry out under extreme condition (as very low temperature), production cost is high, and needs to use a large amount of poisonous and hazardous chemical solvents, can cause very large harm to environmental and human health impacts.
Enzyme process obtains 7-ACA by cephalosporin C acrylase cracking cephalosporin; it is little that production process has floor space; environmental pollution is little; lower-cost feature; but the content of the 7-ACA that current used enzyme preparation technique obtains is lower; look level is higher, could be used for the production of cephalosporins after needing to carry out purifying, so the space be still improved.
Summary of the invention
The object of this invention is to provide a kind of enzyme preparation technique of 7-amino-cephalosporanic acid, the 7-amino-cephalosporanic acid content prepared by this technique is more than 99%, and look level is 2#, can be directly used in the production of cephalosporins without the need to purifying.
In order to realize object of the present invention, inventor provide following technical scheme.
An enzyme preparation technique for 7-amino-cephalosporanic acid, comprises following operation steps:
A. in cephalosporin concentrated solution, drop into cephalosporin C acrylase, control feed temperature is 15-25 DEG C, adds ammoniacal liquor, until the pH value of feed liquid is stabilized in 8.2-8.5;
B. the feed liquid after pH value being stablized uses ultrafiltration membrance filter, and gained filtrate is for subsequent use;
C. the temperature controlling filtrate is 10-12 DEG C, makes it be filtered by ultra-filtration membrane, obtains secondary filtrate, for subsequent use;
D. in secondary filtrate, add dilute hydrochloric acid, regulate its pH value to be 3.8-4.0, filter, filter cake is through washing, dry, namely obtains 7-amino-cephalosporanic acid.
The enzyme preparation technique of above-mentioned 7-amino-cephalosporanic acid, in concentrated solution described in steps A, cephalosporin tires as 15000-35000 μ g/ml.The cracking of enzyme is suitable within the scope of this.
The amount ratio of cephalosporin described in steps A and cephalosporin C acrylase is 22.5-52.5g:50-80U.
The concentration of ammoniacal liquor described in steps A is 10-15%.The speed that adds of ammoniacal liquor is 100-120ml/40min.
Filter described in step B and use ultra-filtration membrane filtering reacted cephalosporin C acrylase bacterium slag, adopt polyethersulfone rolling ultrafiltration membrane, molecular weight cut-off is 5-10 ten thousand dalton.
Ultra-filtration membrane described in step C is polyethersulfone rolling ultrafiltration membrane.Ultra-filtration membrane molecular weight cut-off is 1500 dalton, can filter out the macromole impurity such as the intracellular toxin in feed liquid, albumen and pigment.The pressure filtered is 2-2.5bar.
The concentration of dilute hydrochloric acid described in step D is 10-15%.The speed that dilute hydrochloric acid adds is 100-120ml/60min.
Washing, dry process conveniently operate.
Cephalosporin concentrated solution of the present invention refers to the solution described in industry routine, namely the cynnematin fermented liquid that obtains of fermentation method successively after filtration, the solution that obtains after concentrating of macroporous resin extraction, nanofiltration.
Enzyme preparation technique of the present invention is decoloured without the need to using gac, by the filtration of ultra-filtration membrane after cracking, in conjunction with the accurate control of each operation steps parameter, finally achieves the lifting of finished product 7-amino-cephalosporanic acid quality.
Embodiment
Below in conjunction with specific embodiment, content of the present invention is described in further detail.
Embodiment 1
The cephalosporin concentrated solution of 1.5L, cephalosporin concentration is 25000 μ g/ml, drops into cephalosporin C acrylase 70U wherein; temperature control 20 DEG C, adds the ammoniacal liquor adjust ph of 10%, and the speed that ammoniacal liquor adds is 110ml/40min; until the pH value of feed liquid is stabilized in 8.3, stop adding ammoniacal liquor.
Feed liquid after pH value being stablized crosses polyethersulfone rolling ultrafiltration membrane; ultra-filtration membrane molecular weight cut-off is 100,000 dalton; the gained filtrate cephalosporin C acrylase bacterium slag that filtering is reacted; it is 10-12 DEG C that the filtrate obtained controls its temperature; cross polyethersulfone rolling ultrafiltration membrane; ultra-filtration membrane molecular weight cut-off is 1500 dalton, and during filtration, pressure is 2.3bar, obtains secondary filtrate.
In secondary filtrate, add the dilute hydrochloric acid adjust ph of 15%, the speed that dilute hydrochloric acid adds is 110ml/60min, and adjust ph is 3.9, have solid to separate out, filter, filter cake washes twice through proper amount of acetone, 60-65 DEG C of dry 30min, obtains 7-amino-cephalosporanic acid 19.9g, yield 53.1%.
Embodiment 2
The cephalosporin concentrated solution of 2L, concentration is 15000 μ g/ml, drops into cephalosporin C acrylase 50U wherein; temperature control 25 DEG C, adds the ammoniacal liquor adjust ph of 15%, and the speed that ammoniacal liquor adds is 100ml/40min; until the pH value of feed liquid is stabilized in 8.2, stop adding ammoniacal liquor.
Feed liquid after pH value being stablized crosses polyethersulfone rolling ultrafiltration membrane; ultra-filtration membrane molecular weight cut-off is 50,000 dalton; the gained filtrate cephalosporin C acrylase bacterium slag that filtering is reacted; it is 10-12 DEG C that the filtrate obtained controls its temperature; cross polyethersulfone rolling ultrafiltration membrane; ultra-filtration membrane molecular weight cut-off is 1500 dalton, and during filtration, pressure is 2.2bar, obtains secondary filtrate.
In secondary filtrate, add the dilute hydrochloric acid adjust ph of 13%, the speed that dilute hydrochloric acid adds is 100ml/60min, and adjust ph is 3.8, have solid to separate out, filter, filter cake washs three times through proper amount of acetone, 60-65 DEG C of dry 40min, obtains 7-amino-cephalosporanic acid 15.9g, yield 53.0%.
Embodiment 3
The cephalosporin concentrated solution of 2L, concentration is 35000 μ g/ml, drops into cephalosporin C acrylase 80U wherein; temperature control 15 DEG C, adds the ammoniacal liquor adjust ph of 12%, and the speed that ammoniacal liquor adds is 120ml/40min; until the pH value of feed liquid is stabilized in 8.5, stop adding ammoniacal liquor.
Feed liquid after pH value being stablized crosses polyethersulfone rolling ultrafiltration membrane; ultra-filtration membrane molecular weight cut-off is 100,000 dalton; the gained filtrate cephalosporin C acrylase bacterium slag that filtering is reacted; it is 10-12 DEG C that the filtrate obtained controls its temperature; cross polyethersulfone rolling ultrafiltration membrane; ultra-filtration membrane molecular weight cut-off is 1500 dalton, and during filtration, pressure is 2.5bar, obtains secondary filtrate.
In secondary filtrate, add the dilute hydrochloric acid adjust ph of 10%, the speed that dilute hydrochloric acid adds is 120ml/60min, and adjust ph is 4.0, have solid to separate out, filter, filter cake washes twice through proper amount of acetone, 60-65 DEG C of dry 35min, obtains 7-amino-cephalosporanic acid 37.0g, yield 52.9%.
Embodiment 4
Contriver has carried out quality examination to the 7-amino-cephalosporanic acid that embodiment 1-3 prepares, and result is as shown in table 1:
Table 1
| Sample | Content | Look level | Intracellular toxin | Albumen | Polymkeric substance | D-7-ACA | DO-7-ACA |
| Embodiment 1 | 99.7％ | 2# | 0.41EU/mg | 3500ppm | 0.22％ | 0.31％ | 0.35％ |
| Embodiment 2 | 99.5％ | 2# | 0.62EU/mg | 4200ppm | 0.30％ | 0.41％ | 0.38％ |
| Embodiment 3 | 99.2％ | 2# | 0.55EU/mg | 4800ppm | 0.29％ | 0.42％ | 0.43％ |
As can be seen from Table 1, prepare 7-amino-cephalosporanic acid content according to the method for the invention and can reach more than 99%, look level can reach 2#, intracellular toxin, albumen, polymkeric substance, accompanying impurities D-7-ACA and DO-7-ACA content are all lower, the production of cephalosporins can be directly used in as intermediate, without the need to carrying out purifying again, and product prepared by the 7-ACA that the quality level of product prepared therefrom in These parameters is all produced higher than other method.
Contriver also investigates the stability of the 7-amino-cephalosporanic acid that embodiment 1-3 prepares, specific as follows:
Sample is deposited 36h under the condition of temperature 30 DEG C, humidity 60%, and detect, result is as shown in table 2.
Table 2
| Sample | Content | Look level | Intracellular toxin | Albumen | Polymkeric substance | D-7-ACA | DO-7-ACA |
| Embodiment 1 | 99.1％ | 3# | 0.44EU/mg | 3500ppm | 0.26％ | 0.38％ | 0.39％ |
| Embodiment 2 | 98.9％ | 3# | 0.61EU/mg | 4200ppm | 0.30％ | 0.49％ | 0.42％ |
| Embodiment 3 | 98.4％ | 3# | 0.56EU/mg | 4800ppm | 0.30％ | 0.47％ | 0.49％ |
Claims (10)
1. an enzyme preparation technique for 7-amino-cephalosporanic acid, is characterized in that, comprises following operation steps:
A. in cephalosporin concentrated solution, drop into cephalosporin C acrylase, control feed temperature is 15-25 DEG C, adds ammoniacal liquor, until the pH value of feed liquid is stabilized in 8.2-8.5;
B. the feed liquid after pH value being stablized uses ultrafiltration membrance filter, and gained filtrate is for subsequent use;
C. the temperature controlling filtrate is 10-12 DEG C, makes it be filtered by ultra-filtration membrane, obtains secondary filtrate, for subsequent use;
D. in secondary filtrate, add dilute hydrochloric acid, regulate its pH value to be 3.8-4.0, filter, filter cake is through washing, dry, namely obtains 7-amino-cephalosporanic acid.
2. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, in concentrated solution described in steps A, cephalosporin tires as 15000-35000 μ g/ml.
3. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, the amount ratio of cephalosporin described in steps A and cephalosporin C acrylase is 22.5-52.5g:50-80U.
4. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, the concentration of ammoniacal liquor described in steps A is 10-15%.
5. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, the speed that adds of ammoniacal liquor described in steps A is 100-120ml/40min.
6. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, ultra-filtration membrane described in step C is polyethersulfone rolling ultrafiltration membrane.
7. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, ultra-filtration membrane molecular weight cut-off described in step C is 1500 dalton.
8. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, the pressure filtered described in step C is 2-2.5bar.
9. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, the concentration of dilute hydrochloric acid described in step D is 10-15%.
10. the enzyme preparation technique of a kind of 7-amino-cephalosporanic acid according to claim 1, is characterized in that, the speed that dilute hydrochloric acid described in step D adds is 100-120ml/60min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510127375.9A CN104651439A (en) | 2015-03-23 | 2015-03-23 | Enzymatic preparation process of 7-aminocephalosporanic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510127375.9A CN104651439A (en) | 2015-03-23 | 2015-03-23 | Enzymatic preparation process of 7-aminocephalosporanic acid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104651439A true CN104651439A (en) | 2015-05-27 |
Family
ID=53243079
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510127375.9A Pending CN104651439A (en) | 2015-03-23 | 2015-03-23 | Enzymatic preparation process of 7-aminocephalosporanic acid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104651439A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107868812A (en) * | 2017-12-20 | 2018-04-03 | 焦作健康元生物制品有限公司 | A kind of improvement production technology of 7 amino-cephalo-alkanoic acid |
| CN109553628A (en) * | 2017-09-25 | 2019-04-02 | 联邦制药(内蒙古)有限公司 | A method of eliminating 7-ACA finished foam |
| CN112625053A (en) * | 2020-12-30 | 2021-04-09 | 伊犁川宁生物技术股份有限公司 | 7-ACA with low content of maximum unknown simple impurity and preparation method thereof |
| CN113150010A (en) * | 2021-04-19 | 2021-07-23 | 瑞阳制药股份有限公司 | 7-aminocephalosporanic acid purification process |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102827912A (en) * | 2012-08-31 | 2012-12-19 | 山东鲁抗立科药业有限公司 | Technology for preparing medicine intermediate D-7-ACA by two enzyme carriers one-step method |
| CN103014114A (en) * | 2012-12-27 | 2013-04-03 | 华北制药河北华民药业有限责任公司 | Method for preparing 7-aminocephalosporanic acid via enzymic method |
| CN103555807A (en) * | 2013-10-31 | 2014-02-05 | 哈药集团制药总厂 | Method for preparing 7-ACA (aminocephalosporanic acid) and obtaining alpha-aminoadipic acid by one-step enzymatic reaction |
| CN104402905A (en) * | 2014-10-30 | 2015-03-11 | 华北制药河北华民药业有限责任公司 | Method for recovering 7-aminocephalosporanic acid (7-ACA) from 7-ACA mother liquor |
-
2015
- 2015-03-23 CN CN201510127375.9A patent/CN104651439A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102827912A (en) * | 2012-08-31 | 2012-12-19 | 山东鲁抗立科药业有限公司 | Technology for preparing medicine intermediate D-7-ACA by two enzyme carriers one-step method |
| CN103014114A (en) * | 2012-12-27 | 2013-04-03 | 华北制药河北华民药业有限责任公司 | Method for preparing 7-aminocephalosporanic acid via enzymic method |
| CN103555807A (en) * | 2013-10-31 | 2014-02-05 | 哈药集团制药总厂 | Method for preparing 7-ACA (aminocephalosporanic acid) and obtaining alpha-aminoadipic acid by one-step enzymatic reaction |
| CN104402905A (en) * | 2014-10-30 | 2015-03-11 | 华北制药河北华民药业有限责任公司 | Method for recovering 7-aminocephalosporanic acid (7-ACA) from 7-ACA mother liquor |
Non-Patent Citations (1)
| Title |
|---|
| 范英敏 等: "超滤膜用于酶法7-ACA脱色中的试验研究", 《煤炭与化工》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109553628A (en) * | 2017-09-25 | 2019-04-02 | 联邦制药(内蒙古)有限公司 | A method of eliminating 7-ACA finished foam |
| CN109553628B (en) * | 2017-09-25 | 2021-03-09 | 联邦制药(内蒙古)有限公司 | Method for eliminating 7-ACA finished product foam |
| CN107868812A (en) * | 2017-12-20 | 2018-04-03 | 焦作健康元生物制品有限公司 | A kind of improvement production technology of 7 amino-cephalo-alkanoic acid |
| CN112625053A (en) * | 2020-12-30 | 2021-04-09 | 伊犁川宁生物技术股份有限公司 | 7-ACA with low content of maximum unknown simple impurity and preparation method thereof |
| CN113150010A (en) * | 2021-04-19 | 2021-07-23 | 瑞阳制药股份有限公司 | 7-aminocephalosporanic acid purification process |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104651439A (en) | Enzymatic preparation process of 7-aminocephalosporanic acid | |
| CN110423241B (en) | Process for preparing cephalosporin C sodium salt and 7-aminocephalosporanic acid | |
| KR100293172B1 (en) | Streptomyces sp. Novel method for isolating clavulanic acid and its pharmaceutically acceptable salts from fermentation broth of P6621 FERM P2804 | |
| CN102863519B (en) | Refining method for vancomycin hydrochloride | |
| CN102925526B (en) | Preparation method for 6-amino penicillanic acid | |
| CN110283869B (en) | Preparation method of 7-aminocephalosporanic acid | |
| CN104278071B (en) | The extracting method of cephalosporin | |
| CN104262431A (en) | Method and device for extracting erythromycin thiocyanate | |
| CN111171097B (en) | Separation and purification method for producing adenosine by fermentation | |
| CN104262435B (en) | A kind of production technology of CAMP | |
| CN104066747A (en) | Process of purification of teicoplanin | |
| TW202012433A (en) | A preparation method of precursor recombinant human insulin or analog thereof | |
| CN102838624B (en) | Method for purifying clavulanic acid from fermentation liquor | |
| CN102812050B (en) | The method of purification of hyaluronic acid and/or its salt | |
| CN111574639B (en) | Method for separating and purifying nitraria tangutorum bobr polysaccharide | |
| CN106496022B (en) | A method of extracting pyruvic acid from microbial fermentation solution or enzymatic conversion liquid | |
| CN105111285A (en) | Daptomycin extraction method | |
| US6110699A (en) | Process for producing 6-amino-penicillanic acid and phenylacetic acid | |
| CN111777680A (en) | Separation and purification process for improving stability of recombinant collagen solution | |
| CN110804078A (en) | Deep decolorization and purification method of glycerol glucoside | |
| CN110305925A (en) | Preparation process of enzyme modified rutin mixture | |
| CN206414979U (en) | A kind of clavulanic acid decolourizes and enrichment facility | |
| CN111848644B (en) | Method for treating penicillin fermentation liquor | |
| CN204162637U (en) | A kind of Matachrom extraction element | |
| CN104593351B (en) | A kind of method for preparing cephalosporin C acrylase immobilised enzymes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150527 |
|
| RJ01 | Rejection of invention patent application after publication |