CN102344426A - Method for extracting and purifying lovastatin - Google Patents
Method for extracting and purifying lovastatin Download PDFInfo
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- CN102344426A CN102344426A CN2010102420447A CN201010242044A CN102344426A CN 102344426 A CN102344426 A CN 102344426A CN 2010102420447 A CN2010102420447 A CN 2010102420447A CN 201010242044 A CN201010242044 A CN 201010242044A CN 102344426 A CN102344426 A CN 102344426A
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Abstract
The invention discloses a method for extracting and purifying lovastatin. The method comprises the following steps of: acidizing the pH of a fermentation liquor for producing lovastatin to 1-3; filtering or centrifugally separating; collecting bacterium slags; extracting the bacterium slags with an organic solvent; washing an extracting solution with an alkaline aqueous solution, an acid aqueous solution and deionized water in sequence; concentrating the washed extracting solution, and crystalizing; separating and drying the crystal to obtain a crude product; and recrystallizing the crude product to obtain purified lovastatin. Due to the adoption of the method, the problem of emulsion during the extraction of lovastatin is solved, and the dissolubility of a lovastatin product in acetone, chloroform and methanol is increased.
Description
Technical field
The present invention relates to the preparation method of bulk drug, particularly from fermented liquid, extract the method for refining lovastatin, belong to field of medicaments.
Background technology
Lovastatin (Lovastatin) is microbe-derived 3-hydroxy-3-methylglutaryl coenzyme A reductase suppressor factor class anticholesteremic agent, and because of its mechanism of action is clear and definite, clinical efficacy is remarkable, is present important clinically control cardiovascular and cerebrovascular diseases medicament.Lovastatin is the secondary metabolite of fungi; Main generation bacterium has terreus (Aspergillus terreus), Penicillium citrinum (Penicillium citrinum) and monascus (Monascus ruber) in the industry; The existing method that prepare lovastatin of extracting mainly is that fermented liquid heating, alkalization are filtered, and then filtrating is extracted or filtrating is carried out the resins exchange extraction.
The lovastatin process for extracting that US Patent No. 2003215932 (A1) is set forth is that fermented liquid is also heated with sulfuric acid acidation, at high temperature stirs cyclisation, then to the fermented liquid extracted in toluene; Filter residue is with soda solution and no brine wash; Carry out vacuum concentration then, low temperature crystallization separates, washing.The toluene consumption is big in this invention, because its huge toxicity is brought very big pressure to environmental protection, the employee's on the production line health receives very big threat, is unfavorable for big production.
PCT patent application WO 9720834 has set forth a kind of lovastatin process for extracting; Fermented liquid is transferred to alkalescence with NaOH; Under agitation extract with toluene and ethanol mixed solvent then; In extraction liquid, add a large amount of toluene and perlite; Use the nitric acid acidifying, add chloroform, press filtration under stirring; Concentrate, crystallization gets crude product recrystallization again.This method is used mixed solvent, reclaims difficulty, in the extraction emulsification serious, environmental protection pressure is big, simultaneously owing to use multiple solvent to make production operation become complicated.
In order to remedy the deficiency of above-mentioned technology; At PCT patent application WO 0063411 a kind of novel method is provided: transfer fermented liquid to alkalescence with NaOH; Stir at low temperatures; Centrifugation; Collect supernatant and transfer to acidity with sulfuric acid; The mixed solvent extraction that adds butyl ester and octane, centrifugation, extraction liquid vacuum concentration low temperature crystallization.Still use the mixed solvent extraction in this invention, reclaim difficulty, alkaline extraction emulsification is serious, needs to use a large amount of emulsion splitters, has brought difficulty for the treating process of back.
Further, the method that adopts among the United States Patent (USP) NO.5712130 is: transfer fermented liquid to acid with hydrochloric acid, add the butyl ester extraction after the cooling, the vacuum concentration extraction liquid is at low temperature crystallization.But still there are some shortcomings in this method: extraction solvent consumption is big, is prone to emulsification, and the purifying of extraction process is not enough, only leans on the recrystallization of back to purify, product poorly soluble.
One Chinese patent application prospectus CN 1583736A provides a kind of method of extracting lovastatin in the fermented liquid with resin: fermented liquid is alkalized; Filter; Adsorb with resin; Water mixed liquid with ethanol, acetone and other organic solvent carries out wash-out; The elutriant condensing crystal gets crude product, carries out recrystallization then.This method also exists big not enough: equipment is many, and area occupied is big, and spissated material liquid volume is huge, and energy consumption is big; Wash-out reclaims difficulty with organic solvent; Product gas purity is mainly controlled through recrystallization, and quality product can not get effective assurance aborning.
To sum up, mainly there is following shortcoming in existing lovastatin process for extracting:
1, extraction liquid or effluent volume are big, in the leaching process emulsification serious, production operation is complicated;
2, quality control mainly concentrates in the last recrystallization processing, and difficult quality is effectively guaranteed;
3, product is poorly soluble, is carrying out when semi-synthetic, is prone to emulsification, has a strong impact on yield.
Summary of the invention
The purpose of this invention is to provide a kind of from the broth extraction that is rich in lovastatin and the method for purifying lovastatin; In an emulsification difficult problem in solving the lovastatin extraction, the raising yield; Strengthen the material purifying of pilot process; Improve final product quality; Solve the ubiquitous poorly soluble problem of lovastatin product, improve the solvability of lovastatin product in acetone, chloroform, methyl alcohol.
For realizing above-mentioned purpose, the present invention adopts following technical scheme:
A kind of also method of purifying lovastatin of extracting may further comprise the steps:
1) fermented liquid that will produce lovastatin is acidified to pH1～3, filters then or centrifugation, collects the bacterium slag;
2) with organic solvent extraction bacterium slag, extraction liquid is used alkaline aqueous solution, acidic aqueous solution and deionized water wash successively;
3) concentrated, the crystallization of the extraction liquid after the washing separates and dried crystals obtains crude product;
4), obtain the lovastatin of purifying to the crude product recrystallization.
In the acidization of step 1) fermented liquid; Normally fermented liquid is acidified to pH1～3 with HCl; Stirred heavyization 1-2 hour; Intracellular lovastatin is retained in the spore; The outer lovastatin acid of spore is fully precipitated, guarantee in the filtrate filtered or in the waste water after centrifugal lovastatin unit in 100u/ml.
Collect the bacterium slag through filtering perhaps centrifugation after the acidifying, filtration can use traditional plate and frame to filter, and centrifugal available separating machine separates, and bacterium slag automatic transport to extraction process, is reduced labour intensity, the raising yield.
Step 2) in the extraction treatment; Calculate consumption of organic solvent according to lovastatin unit content in the fermented liquid; By lovastatin content in the extraction liquid is that 3-4 ten thousand units/ml adds water-insoluble organic solvent; A kind of in toluene, butyl ester, ethyl ester and the chloroform for example; Under temperature 50-60 ℃ condition, stir extraction 3-6 hour; With carrying out vacuum filtration under separating machine separation or the air tight condition, collect organic extract liquid, extraction liquid is washed.
Preferred washing process is: extraction liquid earlier with the 0.5-1.0mol/LNaOH of extraction liquid volume 20-30% (V/V) under temperature 30-40 ℃ condition agitator treating 30-50 minute, is left standstill after 15-30 minute again and removes water; The 0.5-1mol/L HCl that uses extraction liquid volume 20-30% (V/V) then under temperature 30-40 ℃ condition agitator treating 30-50 minute leaves standstill after 15-30 minute again and removes water; The no salt solution of using extraction liquid volume 20-30% (V/V) at last under temperature 30-40 ℃ condition agitator treating 30-50 minute leaves standstill after 15-30 minute again and removes water.
Extraction liquid after the step 3) washing concentrates under temperature 60-70 ℃, vacuum 0.075-0.1MPa condition, and the content of lovastatin is controlled at 25-40 ten thousand units/ml in the concentrated solution; Concentrated solution was cooled to 10-15 ℃ of crystallization 3-10 hour, and centrifugation gets damp crystalline substance; Tide is brilliant hour to obtain crude product at temperature 50-60 ℃, vacuum 0.09-01MPa condition following time of drying of 3-5.
During step 4) is handled the crude product recrystallization; A kind of dissolution with solvents crude product in available ethanol, n-propyl alcohol, Virahol, propyl carbinol, isopropylcarbinol, acetone, butyl ester and the ethyl acetate; Concentration is controlled at 0.2-0.5g/ml; Temperature 60-75 ℃; And the gac of adding crude product weight 0.3-0.6%, the dissolving crude product after-filtration, filtrating slowly cools to 10-15 ℃; Be incubated 3-10 hour and carry out recrystallization, centrifugation and drying obtain the pure article of lovastatin.
The invention provides a kind of also novel method of purifying lovastatin of from the fermented liquid that is rich in lovastatin, extracting, improved the quality of lovastatin comprehensively.Extract purification techniques relatively with existing lovastatin, the present invention has adopted elder generation that the great amount of wastewater in the fermented liquid is leached, and has reduced the volume of extraction liquid, has eliminated the emulsification of extraction process; Through the reinforced washing to extraction liquid, having removed influences the deliquescent impurity of lovastatin, has improved the quality of intermediate material greatly, and acid, alkali number that washing process uses are equal basically, and the back discharging that can neutralize has voluntarily reduced environmental protection pressure; Extraction, recrystallization process all use single solvent, help reclaiming.The lovastatin purity of present method preparation is more than 99.5%, and maximum list is planted foreign matter content＜0.15% (weight percent), and generally about 0.1%, product is dissolving fully in acetone, chloroform, methyl alcohol, and the solution clarification is suitable for the industrialization continuous production.
Embodiment
Embodiment 1
Fermented liquid 10L; The 8458u/ml of lovastatin unit transfers to pH1.5 with 5mol/L HCl, stirs heavyization 1 hour; Filter; Collect the bacterium slag,, under 55 ℃ of conditions of temperature, stirred extraction bacterium slag 3 hours in the bacterium slag, adding the 2.5L butyl ester; Separate; Collect extraction liquid, insulation adds 500ml 0.5mol/L NaOH agitator treating 30 minutes at 30 ℃; Leave standstill and removed water in 15 minutes; Organic phase adds 500ml 0.5mol/L HCl, and agitator treating 30 minutes leaves standstill and removed water in 15 minutes; Organic phase adds the 500ml deionized water; Agitator treating 30 minutes leaves standstill and removed water in 15 minutes, and organic phase is carried out (temperature 65-70 ℃ of vacuum concentration; Vacuum tightness is less than 0.08MPa) to the about 300ml of volume; Cool to 12 ℃, be incubated 3 hours, centrifugation and the dry crude product 71g that gets.
Crude product 71g adds 250ml ethanol, and thermosol under 70 ℃ of conditions of temperature adds the 0.3g gac, filters, and slowly cools to 12 ℃ of crystallizations, is incubated 3 hours, and centrifugal drying gets elaboration 60g.Elaboration lovastatin content 99.6%, single foreign matter content of planting is below 0.12%.
Embodiment 2
Fermented liquid 10L; The 7850u/ml of lovastatin unit transfers to pH1.0 with 5mol/L HCl, stirs heavyization 1 hour; Filter; Collect the bacterium slag, in the bacterium slag, add the 2.5L butyl ester, under 54 ℃ of conditions of temperature, stirred extraction bacterium slag 3 hours; Separate; Collect extraction liquid, insulation adds 500ml 0.5mol/L NaOH agitator treating 30 minutes at 30 ℃; Leave standstill and removed water in 15 minutes; Organic phase adds 500ml 0.5mol/LHCl, and agitator treating 30 minutes leaves standstill and removed water in 15 minutes; Organic phase adds the 500ml deionized water; Agitator treating 30 minutes leaves standstill and removed water in 15 minutes, and organic phase is carried out (temperature 65-70 ℃ of vacuum concentration; Vacuum tightness is less than 0.08MPa) to the about 280ml of volume; Cool to 14 ℃, be incubated 3 hours, centrifugation and the dry crude product 70g that gets.
Crude product 70g adds 210ml ethanol, and thermosol under 71 ℃ of conditions of temperature adds the 0.3g gac, filters, and slowly cools to 13 ℃ of crystallizations, is incubated 3 hours, and centrifugal drying gets elaboration 55g.Elaboration lovastatin content 99.5%, single foreign matter content of planting is below 0.11%.
More than described lovastatin provided by the present invention through embodiment and extracted and purification process; Those skilled in the art is to be understood that; In the scope that does not break away from essence of the present invention; Can make certain variation or modification to the present invention, so protection scope of the present invention is looked the claim scope and is defined.
Claims (10)
1. one kind is extracted the also method of purifying lovastatin, may further comprise the steps:
1) fermented liquid that will produce lovastatin is acidified to pH1～3, filters then or centrifugation, collects the bacterium slag;
2) with organic solvent extraction bacterium slag, extraction liquid is used alkaline aqueous solution, acidic aqueous solution and deionized water wash successively;
3) extraction liquid after will washing concentrate, crystallization, separate and dried crystals obtains crude product;
4), obtain the lovastatin of purifying to the crude product recrystallization.
2. the method for claim 1 is characterized in that, in the step 1) with the hcl acidifying fermented liquid to pH1～3, and stirred heavyization 1-2 hour.
3. the method for claim 1 is characterized in that, the laggard andante frame of fermented liquid acidifying filters in the step 1), to collect the bacterium slag.
4. the method for claim 1 is characterized in that step 2) in extraction be selected from a kind of in toluene, butyl ester, ethyl ester and the chloroform with organic solvent.
5. the method for claim 1 is characterized in that step 2) the bacterium slag is stirred extraction 3-6 hour with organic solvent under temperature 50-60 ℃ condition, collect extraction liquid afterwards, extraction liquid is washed.
6. the method for claim 1; It is characterized in that; The process of step 2) extraction liquid being washed is: add earlier the 0.5-1.0mol/L NaOH of extraction liquid volume 20-30%, under temperature 30-40 ℃ condition agitator treating 30-50 minute, leave standstill after 15-30 minute and remove water; The 0.5-1mol/L HCl that adds extraction liquid volume 20-30% again under temperature 30-40 ℃ condition agitator treating 30-50 minute, leaves standstill after 15-30 minute and removes water in organic phase; The deionized water that adds extraction liquid volume 20-30% at last under temperature 30-40 ℃ condition agitator treating 30-50 minute, leaves standstill after 15-30 minute and removes water in organic phase.
7. the method for claim 1 is characterized in that, the extraction liquid after will washing in the step 3) concentrates under temperature 60-70 ℃, vacuum 0.075-0.1MPa condition, and the content of lovastatin is controlled at 25-40 ten thousand units/ml in the concentrated solution, carries out crystallization then.
8. the method for claim 1 is characterized in that, the crystalline temperature is 10-15 ℃ in the step 3), and the time is 3-10 hour.
9. the method for claim 1 is characterized in that, step 4) is carried out recrystallization afterwards with a kind of dissolution with solvents crude product in ethanol, n-propyl alcohol, Virahol, propyl carbinol, isopropylcarbinol, acetone, butyl ester and the ethyl acetate.
10. method as claimed in claim 9 is characterized in that, step 4) is dissolved crude product down at temperature 60-75 ℃; And in solution, add the gac of crude product weight 0.3-0.6%; The dissolving crude product after-filtration, filtrating slowly is cooled to 10-15 ℃ and carries out recrystallization, obtains the pure article of lovastatin.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2010102420447A CN102344426A (en) | 2010-07-30 | 2010-07-30 | Method for extracting and purifying lovastatin |
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| CN2010102420447A CN102344426A (en) | 2010-07-30 | 2010-07-30 | Method for extracting and purifying lovastatin |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102827123A (en) * | 2012-08-02 | 2012-12-19 | 丽珠集团新北江制药股份有限公司 | Separation and extraction process for Mevastatin |
| CN104341377A (en) * | 2013-08-05 | 2015-02-11 | 北大方正集团有限公司 | Method for recovering lovastatin from lovastatin crystallization mother liquor |
| CN106083789A (en) * | 2016-06-07 | 2016-11-09 | 山东鲁抗医药股份有限公司 | The method that precipitate and separate extracts lovastatin from fermentation liquid |
| CN106496174A (en) * | 2016-09-30 | 2017-03-15 | 丽珠集团宁夏新北江制药有限公司 | A kind of extraction and purification process of Lovastatin |
| CN106518824A (en) * | 2016-09-30 | 2017-03-22 | 丽珠集团宁夏新北江制药有限公司 | Extraction technology of lovastatin |
| CN108976190A (en) * | 2017-12-21 | 2018-12-11 | 北大方正集团有限公司 | A method of recycling Lovastatin from Lovastatin crystalline mother solution |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994029292A1 (en) * | 1993-06-08 | 1994-12-22 | Krka Tovarna Zdravil P.O. | Process for the isolation of lovastatin |
| US20030215932A1 (en) * | 2000-06-30 | 2003-11-20 | Parveen Kumar | Process for the isolation of lovastatin |
| CN1583736A (en) * | 2004-06-03 | 2005-02-23 | 山东鲁抗医药股份有限公司 | Extraction and refinement of lovastatin |
| US20090156837A1 (en) * | 2007-12-18 | 2009-06-18 | Themis Medicare Limited | Isolation and recovery of simvastatin in lactone form or in the form of an acid salt from the harvested fermentation broth |
-
2010
- 2010-07-30 CN CN2010102420447A patent/CN102344426A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994029292A1 (en) * | 1993-06-08 | 1994-12-22 | Krka Tovarna Zdravil P.O. | Process for the isolation of lovastatin |
| US20030215932A1 (en) * | 2000-06-30 | 2003-11-20 | Parveen Kumar | Process for the isolation of lovastatin |
| CN1583736A (en) * | 2004-06-03 | 2005-02-23 | 山东鲁抗医药股份有限公司 | Extraction and refinement of lovastatin |
| US20090156837A1 (en) * | 2007-12-18 | 2009-06-18 | Themis Medicare Limited | Isolation and recovery of simvastatin in lactone form or in the form of an acid salt from the harvested fermentation broth |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102827123A (en) * | 2012-08-02 | 2012-12-19 | 丽珠集团新北江制药股份有限公司 | Separation and extraction process for Mevastatin |
| CN102827123B (en) * | 2012-08-02 | 2015-04-22 | 丽珠集团新北江制药股份有限公司 | Separation and extraction process for Mevastatin |
| CN104341377A (en) * | 2013-08-05 | 2015-02-11 | 北大方正集团有限公司 | Method for recovering lovastatin from lovastatin crystallization mother liquor |
| CN104341377B (en) * | 2013-08-05 | 2016-10-19 | 北大方正集团有限公司 | The method reclaiming lovastatin from lovastatin crystalline mother solution |
| CN106083789A (en) * | 2016-06-07 | 2016-11-09 | 山东鲁抗医药股份有限公司 | The method that precipitate and separate extracts lovastatin from fermentation liquid |
| CN106496174A (en) * | 2016-09-30 | 2017-03-15 | 丽珠集团宁夏新北江制药有限公司 | A kind of extraction and purification process of Lovastatin |
| CN106518824A (en) * | 2016-09-30 | 2017-03-22 | 丽珠集团宁夏新北江制药有限公司 | Extraction technology of lovastatin |
| CN106496174B (en) * | 2016-09-30 | 2019-06-04 | 丽珠集团(宁夏)制药有限公司 | A kind of extraction and purification process of Lovastatin |
| CN106518824B (en) * | 2016-09-30 | 2019-06-04 | 丽珠集团(宁夏)制药有限公司 | A kind of extraction process of Lovastatin |
| CN108976190A (en) * | 2017-12-21 | 2018-12-11 | 北大方正集团有限公司 | A method of recycling Lovastatin from Lovastatin crystalline mother solution |
| CN108976190B (en) * | 2017-12-21 | 2020-09-04 | 北大方正集团有限公司 | Method for recovering lovastatin from lovastatin crystallization mother liquor |
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Application publication date: 20120208 |