CN102086226B - Method for preparing cyclosporine A - Google Patents
Method for preparing cyclosporine A Download PDFInfo
- Publication number
- CN102086226B CN102086226B CN 200910253905 CN200910253905A CN102086226B CN 102086226 B CN102086226 B CN 102086226B CN 200910253905 CN200910253905 CN 200910253905 CN 200910253905 A CN200910253905 A CN 200910253905A CN 102086226 B CN102086226 B CN 102086226B
- Authority
- CN
- China
- Prior art keywords
- cyclosporin
- purification
- fermented liquid
- separation
- silica gel
- 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.)
- Active
Links
Images
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to the field of biomedical technology and provides an industrialized method for preparing high-purity cyclosporine A. According to the method, the cyclosporine A is obtained through extraction by utilizing fermentation liquid, chromatography by utilizing a nonpolar macroporous absorption resin, silica gel column chromatography, crystallization and purification. In the invention, isoconcentration elution is adopted without complicated intermediary steps of dissolvent substitution processes; the process is easy to operate and is suitable for industrialized production; the purity of products is greater than 98.5; and the yield is above 65%.
Description
Technical field
The present invention relates to the biological medicine technology field, be specifically related to a kind of separation purifying technique of cyclosporin A.
Background technology
Cyclosporin A (Ciclosporin A; CsA) be from porous wood mould (Tolypocladium inflatum CSP3); After separate obtain a kind of in the culture of snow-white muscardine (Beauveria bassiana) just by name and contain 11 amino acid whose ring type polypeptides; Be the very strong immunosuppressor of a kind of effect, except cyclosporin A, also have more than 20 kind of homologues such as B, C, D.Ciclosporin A is used for the immunosuppressant therapy of transplantations such as kidney, liver and heart; It can also be used to treating some autoimmune diseases, like various autoimmune property diseases such as lupus erythematosus, rheumatoid arthritis, idiopathic thrombocytopenic purpura, ulcerative colitis, myasthenia gravis, chronic nephritis, chronic active hepatitis and nephrotic syndromes.
Major impurity in the cyclosporin A leavened prod is the cyclosporin A homologue of structural similitude; Comprise B, C, D etc.; Extraction purifying main technique route for cyclosporin A in the prior art is to use organic solvent extraction earlier, carries out separation and purification through dissimilar column chromatographies then.
CN200510108097.9 has described a kind of S-Neoral coarse crystal that at first obtains; Utilize the column chromatographic isolation and purification cyclosporin A again, and carry out the purifying process of secondary crystal, this method coarse crystallization complex process; With an organic solvent kind is many; Some solvent price is more expensive, and crystallization needs 3 days, whole purifying process length consuming time.
US5656459 discloses a kind of extraction and purification process; Use earlier methanol extraction, remove methyl alcohol and again extract is dissolved in ETHYLE ACETATE, obtain the pure article of cyclosporin A through twice silica gel column chromatography; This technology is extracted and can not be connected with the silica gel column chromatography solvent for use; Need conversion, and silica gel column chromatography often, total recovery is low.
US5382655 discloses a kind of with chloroform, methylene dichloride and the ethanol silica gel column chromatography method as moving phase separation and purification cyclosporin A, but these solvent toxicity are big, and is unfriendly to environment.Patent US597638 discloses a kind of high-pressure carbon dioxide that utilizes as the method for flowing carrier through silica gel column chromatography acquisition cyclosporin A, but this method is high to equipment requirements, and quantity of sample handling is few, can't realize extensive large-scale production.Patent US2002/0162789A1 discloses and has a kind ofly separated with the multistep silica gel chromatography, and moving phase toluene and ETHYLE ACETATE, main drawback are that toluene toxicity is big, and chromatographic step is many, and solvent consumption is big.WO01/64935A1 has described a kind of method of utilizing organic solvent extraction, gel-filtration and silica gel column chromatography separating purification cyclosporin A; This method main drawback is that the gel-filtration filler costs an arm and a leg; And all need removal of solvents between each step with back, and workload is big, and solvent consumption is many.US2003/0186856A1 adopts the method for polydextran gel through twice chromatographic separation and purification cyclosporin A, and subject matter is the filler expensive, and cost is high.CA2108655 provides a kind of method that adopts the superfluid abstraction technique to separate cyclosporin A, and this method investment is big, and cost is high, is not suitable for suitability for industrialized production.
The principal feature of comprehensive publication is to extract with organic solvent earlier; Utilize a step or multistep silica gel column chromatography to carry out separation and purification then; Some separation purifying technique cost is high, is not suitable for suitability for industrialized production, and some process solvent toxicity is big; Consumption is big, and total recovery about 40%～60%.
" (numbering: 1001-8689 (2008) 05-0276-04) report utilizes macroporous adsorbent resin HP to combine high speed adverse current chromatogram to separate the S-Neoral small component to Chinese microbiotic journal article; Can realize that enrichment separates again to S-Neoral; But this method treatment scale is little, equipment requirements is high, is unfavorable for suitability for industrialized production.
Summary of the invention
The object of the present invention is to provide a kind of method of extensive, efficient preparation of industrialization high purity cyclosporin A.
Complex process to cyclosporin A separation and purification ubiquity in the prior art, complex steps, the organic solvent kind is many and consumption is big; Shortcomings such as pigment poor removal effect and total recovery are low, the contriver carries out a large amount of research to the cyclosporin A separation purifying technique, to the biochemical property of ciclosporin A; Grope experiment condition and processing parameter, the separation method that adopts macroporous absorption and silica gel column chromatography to combine has been obtained the good technical effect; The present invention adopts the macroporous adsorption resin chromatography technology; Organic solvent in the extracting solution need not displacement can directly go up an appearance macroporous adsorbent resin, and the eluent consumption is few, through experiment only confirm need 1.5～2 times of column volumes can be with the complete wash-out of cyclosporin A; Cyclosporin A is played the enrichment method effect of highly significant; And can remove most pigments, solve the shortcoming of original extraction process, improve product yield; Because macroporous adsorption resin chromatography has been removed most of impurity; The present invention adopts non-gradient elution in silica gel column chromatography, and uses low organic solvent-acetone of toxicity and sherwood oil as moving phase, compares gradient elution; Simplify chromatography technology, reduced environmental pollution.
The present invention realizes through following technical scheme:
(1) extraction of cyclosporin A
In fermented liquid, add NaOH and transfer PH, add organic solvent, stir and extracted 5～8 hours, extracting solution is leached thalline, collect filtrating to neutral.
Wherein in fermented liquid, add organic solvent and be in methyl alcohol, ethanol, ETHYLE ACETATE and the acetone one or more; Be preferably acetone; Fermented liquid is 1: 1～1: 2 with the volume of organic solvent ratio; Preferred 1: 1, after stirring 5～8 hours under 30 ℃～40 ℃ conditions, carry out solid-liquid separation and collect filtrating.
(2) macroporous adsorption resin chromatography
Macroporous adsorbent resin dress post carries out pre-treatment to resin, and appearance on the extracting solution is handled the resin of going up after the appearance with washing composition, and elutriant wash-out and collect elutriant adds the petroleum ether extraction elutriant then, concentrates extraction liquid for use.
Different model resin D101, HP20, HPD300, XDA-6 and X-5 have been carried out cyclosporin A Static Adsorption-desorption experiment, confirmed that absorption and the desorption effect of D101, HP20 and X-5 is good, wherein more preferably D101 resin.
When acetone content is lower than 50% in the extracting solution, the D101 resin to the maximal absorptive capacity of cyclosporin A greater than the 30mg/g resin, further test confirm extracting solution adsorptive capacity greater than 10 times of column volumes; The used washing composition of pre-treatment resin is pure water, 3% sodium hydroxide solution and 3% hydrochloric acid soln successively; Resin washing composition after the last appearance is followed successively by 3% hydrochloric acid and pure water; Eluent is one or more in ETHYLE ACETATE, methyl alcohol and the acetone; Preferred acetone, elution volume is about 1.5～2 times of column volumes.
(3) silica gel column chromatography
Silica gel dress post will extract appearance on the liquid concentrator, with acetone sherwood oil mixed solvent wash-out, collect the qualified cyclosporin A elutriant of purity.
Silica gel column chromatography selects for use high order to count silica gel, one or both in 200-300 order or the 100-200 order, preferred 200-300 order; Silica gel column chromatography post aspect ratio scope is 8: 1～12: 1, preferred 9: 1 and 10: 1, and more preferably 10: 1; The silica gel column chromatography elutriant consists of acetone and sherwood oil; Proportioning is an acetone: sherwood oil (volume ratio)=20～40: 60～80, and preferred 40: 60 and 30: 70, more preferably 30: 70; When column volume was 14L, sample solution was 20% column volume, and appearance concentration is 80～110g/L on the cyclosporin A, preferred 100g/L, and eluent flow rate is 70～90mL/min, preferred 80mL/min detects effluent with HPLC, collects specification product.
(4) crystallization purifying
Washing with acetone is used in elutriant vacuum concentration and crystallization, and last vacuum-drying gets the pure article of colourless powder shape cyclosporin A.
Elutriant is evaporated to small volume under 60 ℃～75 ℃ conditions, add the small amount of acetone dissolving, places crystallization under-5 ℃～-20 ℃ conditions, and preferred-10 ℃, filter and collect crystal, wash under back 40 ℃～70 ℃ conditions with cold acetone and be dried to constant weight.
Technique effect of the present invention is: through adopting macroporous adsorption resin chromatography fractionation by adsorption cyclosporin A; With respect to traditional extraction technology; Can make consumption of organic solvent reduce 1 times, pigment removal reaches 95%, and the concentration of cyclosporin A can improve nearly 10 times through enrichment method; Slave unit can save bulky extractor on requiring, and cuts down the consumption of energy and labour intensity; Silica gel column chromatography employing acetone that toxicity is low, price is relatively cheap and sherwood oil are as moving phase; Adopt the isoconcentration wash-out with respect to gradient elution, reduced process step, improved yield (more than 65%) and purity (98.5% above HPLC); See from whole operational path; Do not have complicated intermediate steps solvent exchange process, technological operation is simple and direct, is suitable for suitability for industrialized production.
Description of drawings
Fig. 1, cyclosporin A preparation technology schema
The HPLC collection of illustrative plates of cyclosporin A behind Fig. 2, the macroporous adsorption resin chromatography
The HPLC collection of illustrative plates of cyclosporin A behind Fig. 3, the purification by silica gel column chromatography
The HPLC collection of illustrative plates of Fig. 4, S-Neoral xln
Embodiment
Further describe beneficial effect of the present invention through following examples at present; Embodiment only is used for the purpose of illustration; Do not limit the scope of the invention, conspicuous change and modification that while those of ordinary skills are made according to the present invention are also contained within the scope of the invention.
Embodiment 1
(1) get cyclosporin A fermented liquid 50L, add 5mol/l NaOH and regulate pH to neutral, add 1 times of volume acetone and extracted 8 hours 30 ℃ of stirrings ,-0.06Mpa vacuum filtration is removed thalline, collects filtrating and treats upper prop.
(2) with 5kg D101 macroporous adsorbent resin dress post (Φ 10cm * 120cm), use pure water, 3% sodium hydroxide solution and 3% hydrochloric acid soln carrying out washing treatment resin respectively, after disposing with the extracting solution upper prop of about 10 times of column volumes; Be washed till pH 6.5～7.5 with the salt pickling post of about 2 column volumes 3% and with pure water; Use the acetone wash-out, collect about 1.5 times of column volume elutriants; In elutriant, add the petroleum ether extraction layering of 0.5 times of volume, remove lower floor's water, concentrated upper phase is treated silicagel column.
(3) 200-300 order silica gel is suitably handled back wet method dress post (Φ 10cm * 140cm); With 2 column volume acetone: sherwood oil (volume ratio)=30: 70 balance silicagel columns; After balance finishes with the liquid concentrator upper prop of about 20% column volume; With above-mentioned acetone/sherwood oil mix reagent wash-out, collect purity and be higher than 97% elutriant.
(4) 60 ℃ of vacuum concentration of elutriant are positioned over 12 hours after-filtration of-5 ℃ of crystallizations, and carry out drip washing with cold acetone, and 40 ℃ of vacuum-dryings obtain white amorphous powder finished product to constant weight then, and calculating total recovery is 67.7%.
(1) get cyclosporin A fermented liquid 50L, add 5mol/l NaOH and regulate pH to neutral, add 2 times of volumes methanol and extracted 5 hours 40 ℃ of stirrings ,-0.06Mpa vacuum filtration is removed thalline, collects filtrating and treats upper prop.
(2) with 5kg H103 macroporous adsorbent resin dress post (Φ 10cm * 120cm), use pure water, 3% sodium hydroxide solution and 3% hydrochloric acid soln carrying out washing treatment resin respectively, after disposing with about 8 times of column volume extracting solution upper props; Be washed till pH 6.5～7.5 with the salt pickling post of 1.5 times of column volumes 3% and with pure water; Use eluent ethyl acetate, collect about 2 times of column volume elutriants, in elutriant, add the petroleum ether extraction layering of 0.5 times of volume; Remove lower floor's water, concentrated upper phase is treated silicagel column.
(3) this step is with embodiment 1 step (3).
(4) 65 ℃ of vacuum concentration of elutriant are positioned over-10 ℃ and carry out 12 hours after-filtration of crystallization, and carry out drip washing with cold acetone, and 70 ℃ of drying under reduced pressure obtain white amorphous powder finished product to constant weight, and calculating total recovery is 65.3%.
Embodiment 3
(1) get cyclosporin A fermented liquid 50L, add 5mol/l NaOH and regulate pH to neutral, add 1.5 times of volumes of acetic acid ethyl esters and extracted 6 hours 35 ℃ of stirrings ,-0.06Mpa vacuum filtration is removed thalline, collects filtrating and treats upper prop.
(2) with 5kg X-5 macroporous adsorbent resin dress post (Φ 10cm * 120cm), use pure water, 3% sodium hydroxide solution and 3% hydrochloric acid soln carrying out washing treatment resin respectively, after disposing with about 7 times of column volume extracting solution upper props; Be washed till pH 6.5～7.5 with the salt pickling post of 2 times of column volumes 3% and with pure water; Use the acetone wash-out, collect about 1.5 times of column volume elutriants; In elutriant, add the petroleum ether extraction layering of 0.5 times of volume, remove lower floor's water, concentrated upper phase is treated silicagel column.
(3) this step is with embodiment 1 step (3).
(4) 70 ℃ of vacuum concentration of elutriant are positioned over-20 ℃ and carry out 12 hours after-filtration of crystallization, carry out drip washing with cold acetone; 65 ℃ of drying under reduced pressure obtain white amorphous powder finished product to constant weight, and calculating total recovery is 66.2%.
(1) this step is with embodiment 1 step (1).
(2) with 5kg D101 macroporous adsorbent resin dress post (Φ 10cm * 120cm), use pure water, 3% sodium hydroxide solution and 3% hydrochloric acid soln carrying out washing treatment resin respectively, after disposing with about 12 times of column volume extracting solutions; Be washed till pH 6.5～7.5 with the salt pickling post of 2 column volumes 3% and with pure water; Use methanol-eluted fractions, collect about 1.5 times of column volume elutriants, in elutriant, add the petroleum ether extraction layering of 0.5 times of volume; Remove lower floor's water, concentrated upper phase is treated silicagel column.
(3) 200-300 order silica gel is suitably handled back wet method dress post (Φ 10cm * 140cm); Acetone with 2 column volumes: sherwood oil (volume ratio)=25: 75 balance silicagel columns; With about 20% column volume liquid concentrator upper prop; With above-mentioned acetone sherwood oil mix reagent wash-out, collect purity and be higher than 97% elutriant.
(4) 75 ℃ of vacuum concentration of elutriant are positioned over-10 ℃ and carry out 12 hours after-filtration of crystallization, and carry out drip washing with cold acetone, and 50 ℃ of drying under reduced pressure obtain white amorphous powder finished product to constant weight, and calculating total recovery is 68.5%.
Embodiment 5
(1) this step is with embodiment 1 step (1).
(2) with 5kg D101 macroporous adsorbent resin dress post (Φ 10cm * 120cm), use pure water, 3% sodium hydroxide solution and 3% hydrochloric acid soln carrying out washing treatment resin respectively, after disposing with appearance on about 11 times of column volume extracting solutions; Be washed till pH 6.5～7.5 with the salt pickling post of 2 column volumes 3% and with pure water; Use the acetone wash-out, collect about 1.5 times of column volume elutriants, add the petroleum ether extraction layering of 0.5 times of volume; Remove lower floor's water, concentrated upper phase is treated silicagel column.
(3) 200-300 order silica gel is suitably handled back wet method dress post (Φ 10cm * 140cm); Acetone with 2 column volumes: sherwood oil (volume ratio)=20: 80 balance silicagel columns; With about 20% column volume liquid concentrator upper prop; With above-mentioned acetone sherwood oil mix reagent wash-out, collect purity and be higher than 97% elutriant.
(4) 65 ℃ of vacuum concentration of elutriant are positioned over-20 ℃ and carry out 12 hours after-filtration of crystallization, and carry out drip washing with cold acetone, and 65 ℃ of drying under reduced pressure obtain white amorphous powder finished product to constant weight, and calculating total recovery is 70.1%.
Claims (1)
1. the method for a separation and purification cyclosporin A from fermented liquid, its process comprises following steps:
(1) extraction of cyclosporin A
In fermented liquid, add NaOH and transfer pH value for neutral, add organic solvent, stir and extracted 5 ~ 8 hours, extracting solution is leached microorganism collection filtrating, said organic solvent is one or more in methyl alcohol, ethanol, ETHYLE ACETATE and the acetone;
(2) macroporous adsorption resin chromatography
Use the nonpolar macroporous adsorption resin dress post of model, resin is carried out pre-treatment, appearance on the extracting solution as D101, HP20, X-5 or H103; With the resin after the last appearance of washing composition processing; Elutriant wash-out and collect elutriant adds the petroleum ether extraction elutriant then, concentrates extraction liquid for use;
(3) silica gel column chromatography
Silica gel dress post will extract appearance on the liquid concentrator, with acetone sherwood oil mixed solvent wash-out, collect the qualified cyclosporin A elutriant of purity;
(4) crystallization purifying
Washing with acetone is used in elutriant vacuum concentration and crystallization, and last vacuum-drying gets the pure article of colourless powder shape cyclosporin A.
2, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (1) fermented liquid and volume of organic solvent than being 1:1 ~ 1:2.
3, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (1) fermented liquid and volume of organic solvent than being 1:1.
4, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (1) organic solvent is an acetone.
5, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (1) broth extraction temperature is 30 ℃ ~ 40 ℃.
6, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (2) big pore adsorption resin model is D101.
7, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that the used washing composition of step (2) pre-treatment resin is pure water, 3% sodium hydroxide solution and 3% hydrochloric acid soln successively.
8, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that the resin washing composition that step (2) goes up after the appearance is followed successively by 3% hydrochloric acid and pure water.
9, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (2) eluent is one or more in ETHYLE ACETATE, methyl alcohol and the acetone.
10, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (2) eluent is an acetone.
11, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that silica gel granularity that step (3) silica gel column chromatography is selected for use is one or both in 200-300 order and the 100-200 order.
12, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (3) silica gel column chromatography post aspect ratio is 8:1～12:1.
13, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (3) silica gel column chromatography post aspect ratio 10:1.
14, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (3) sample solution is 10% ~ 30% column volume.
15, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (3) sample solution is 20% column volume.
16, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that the elutriant proportioning is a volume ratio acetone in step (3) silica gel column chromatography: sherwood oil=20～40:60～80.
17, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that the elutriant proportioning is volume ratio acetone: sherwood oil=30:70 in step (3) silica gel column chromatography.
18, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that step (4) silica gel column chromatography elutriant vacuum concentration temperature is 60 ℃ ~ 75 ℃.
19, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that the Tc of step (4) cyclosporin A is-5 ℃～-20 ℃.
20, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, it is characterized in that the Tc of step (4) cyclosporin A is-10 ℃.
21, according to claim 1 a kind of from fermented liquid the method for separation and purification cyclosporin A, the vacuum-drying temperature that it is characterized in that step (4) cyclosporin A is 40 ℃ ~ 70 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200910253905 CN102086226B (en) | 2009-12-04 | 2009-12-04 | Method for preparing cyclosporine A |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200910253905 CN102086226B (en) | 2009-12-04 | 2009-12-04 | Method for preparing cyclosporine A |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102086226A CN102086226A (en) | 2011-06-08 |
| CN102086226B true CN102086226B (en) | 2012-10-10 |
Family
ID=44098248
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200910253905 Active CN102086226B (en) | 2009-12-04 | 2009-12-04 | Method for preparing cyclosporine A |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102086226B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102368072B (en) * | 2011-06-30 | 2014-09-24 | 同昕生物技术（北京）有限公司 | Chemiluminescent enzyme-linked immunoassay kit for detecting concentration of cyclosporine A drug |
| CN102367270B (en) * | 2011-06-30 | 2015-04-01 | 同昕生物技术（北京）有限公司 | Preparation method of cyclosporin A haptin and enzymelinked immunosorbent quantitative detection kit of cyclosporin A |
| CN103073624B (en) * | 2011-10-25 | 2016-02-24 | 华北制药集团新药研究开发有限责任公司 | A kind of preparation method of high purity cyclosporin A derivative |
| CN108250275A (en) * | 2016-12-29 | 2018-07-06 | 天津领世生物科技开发有限公司 | A kind of method that cyclosporin A is isolated and purified from zymotic fluid |
| CN108864257A (en) * | 2018-06-28 | 2018-11-23 | 杭州中美华东制药有限公司 | High purity cyclosporin A and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1212703A (en) * | 1996-03-21 | 1999-03-31 | 德雷斯顿药品工厂有限公司 | Chromatographic process for obtaining very pure cycloporin A and related cyclosporins |
| CN1570130A (en) * | 2004-05-09 | 2005-01-26 | 上海医药工业研究院 | Cyclosporins A fermentation production method |
| CN1763084A (en) * | 2005-10-11 | 2006-04-26 | 山东新时代药业有限公司 | High purity cyclosporin A preparation method |
-
2009
- 2009-12-04 CN CN 200910253905 patent/CN102086226B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1212703A (en) * | 1996-03-21 | 1999-03-31 | 德雷斯顿药品工厂有限公司 | Chromatographic process for obtaining very pure cycloporin A and related cyclosporins |
| CN1570130A (en) * | 2004-05-09 | 2005-01-26 | 上海医药工业研究院 | Cyclosporins A fermentation production method |
| CN1763084A (en) * | 2005-10-11 | 2006-04-26 | 山东新时代药业有限公司 | High purity cyclosporin A preparation method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102086226A (en) | 2011-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ni et al. | Column chromatographic extraction and preparation of cordycepin from Cordyceps militaris waster medium | |
| CN102086226B (en) | Method for preparing cyclosporine A | |
| CN103275152B (en) | A kind of preparation method of high-purity fidaxomicin | |
| CN102936253B (en) | A kind of preparation method of high purity tacrolimus | |
| CN109867663A (en) | A method of with preparation chromatographic isolation chrysomycin A and chrysomycin B | |
| CN102443012B (en) | A kind of method of purifying rapamycin from fermented liquid | |
| CN102070688A (en) | Method for enriching and purifying icariin in epimedium herb | |
| JP6592500B2 (en) | Purification method of fidaxomycin | |
| CN103058871B (en) | Separation and purification method of tobacco chlorogenic acid | |
| CN104844620B (en) | A kind of isolation and purification method of rapamycin | |
| CN108840845A (en) | The method of Xanthatin is extracted from Siberian cocklebur | |
| CN101869581B (en) | Technical method for extracting active component from Chinese herbal medicine | |
| CN113512036A (en) | Method for extracting and separating phellodendrine from phellodendron amurense | |
| CN105585578A (en) | A preparing method of high-purity rapamune | |
| CN104231011B (en) | Preparation method of verbascoside | |
| CN103570647A (en) | Method for preparing high-purity paclitaxel from taxus chinensis cell culture fluid | |
| CN1931835A (en) | Process of extracting citrulline from water melon | |
| CN101759641A (en) | Method for extracting and separating huperzine A from huperizia serrata | |
| CN103073624B (en) | A kind of preparation method of high purity cyclosporin A derivative | |
| CN108250275A (en) | A kind of method that cyclosporin A is isolated and purified from zymotic fluid | |
| CN109609568B (en) | Method for preparing swainsonine by solid fermentation | |
| CN101353294A (en) | Separation and purification method of high-content resveratrol | |
| CN103467352B (en) | A kind of method of prodigiosin extraction purification | |
| CN102321153B (en) | Preparation method of xin'ao glycoside peptide powdery solid | |
| CN105085443A (en) | Method for using Taxus media branches and leaves as raw material to prepare high-purity 10-deacetyl baccatin III |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221123 Address after: 273400 Shandong city of Linyi province Feixian County North Ring Road No. 1 Patentee after: LUNAN NEW TIME BIO-TECH Co.,Ltd. Address before: 273400 Shandong city of Linyi province Feixian County North Ring Road No. 1 Patentee before: SHANDONG NEW TIME PHARMACEUTICAL Co.,Ltd. |
|
| TR01 | Transfer of patent right |