A kind of preparation method of 6-aminopenicillanic acid
Technical field
The present invention relates to the preparation method of pharmaceutical raw material, specifically a kind of preparation method of 6-aminopenicillanic acid.
Background technology
6-aminopenicillanic acid is called for short 6-APA.With 6-APA is parent nucleus, can be prepared into the beta-lactam semisynthetic antibiotics with different side chain condensation.The beta-lactam semisynthetic antibiotics is widely used in clinical because of its determined curative effect, toxic side effect are little.Along with a large amount of uses of a series of beta-lactam semisynthetic antibiotics such as amoxycilline Trihydrate bp, Ampicillin Trihydrate, the market demand of its raw material 6-APA is also in continuous rising.
At present, prepare the method for 6-APA, generally comprise following steps: a, with the penicillin fermentation liquid be raw material after filtration, acidifying, butanols concentration extraction, make penicillin butyl ester extracting solution; B, with penicillin butyl ester extracting solution alkalization, make the penicillin salt aqueous solution, again with the butanols azeotropic, after filtration, washing, crystallization, drying be prepared into potassium salt of penicillin industry powder; C, be raw material,, obtain 6-APA through dissolving, enzymatic conversion, crystallization with potassium salt of penicillin industry powder.In this method processing step two times of crystallization is arranged.This just causes it, and yield losses is many in dual crystallisation process, product yield is low, also causes the waste of solvent, power and energy simultaneously, has strengthened production cost.Prepare 6-APA as stated above, the yield of its finished product is generally 81%.
Summary of the invention
Purpose of the present invention will provide a kind of 6-APA preparation method exactly, and this method production cost is low, the product yield height, and can be used for suitability for industrialized production.
The objective of the invention is such realization: the inventive method may further comprise the steps:
A, with penicillin fermentation liquid filtration, acidifying, butyl alcohol extraction concentrate, decolour penicillin butyl ester extracting solution (being called for short a time BA);
B, penicillin butyl ester extracting solution get the penicillin salt aqueous solution (being called for short RB) through the basic solution back extraction;
C, the penicillin salt aqueous solution carry out degreasing, washed resin post through resin column, are prepared into the degreasing penicillin salt aqueous solution;
After d, degreasing penicillin salt aqueous solution injection penicillin acylase retort are carried out enzymatic conversion, add the 6-APA crystal seed, growing the grain, crystallization, drying.
Processing parameter in a, the b step can carry out according to the processing parameter in the ordinary method.As in the prior art when preparation benzylpenicillin potassium (sodium) salt or the 6-APA, this acidifying all arranged, concentrate, technological processs such as degreasing, alkali lye back extraction.Basic solution can be salt of wormwood, yellow soda ash, but preferred salt of wormwood.
In the c step, the degreasing penicillin salt aqueous solution preferably carries out degreasing with the velocity flow of 200-400L/h through resin column, to remove residual butyl ester in the degreasing penicillin salt aqueous solution better; During the washed resin post, the volume that preferably makes the penicillin salt aqueous solution after the degreasing be the degreasing front volume 1.5-3 doubly; Can effectively prolong the work-ing life of penicillin acylase, further reduce production costs.
Employed resin is preferably ADS-18, AXT-33.Preferred ADS-18 can further guarantee the quality of resins exchange like this.
In the c step, in the retort of penicillin acylase, processing parameter preferably adds borate buffer by the 3%-10% of degreasing penicillin salt aqueous solution cumulative volume, carries out conversion reaction; Keep pH6.5-9.0 with ammoniacal liquor, temperature is controlled at 25-35 ℃, reaction times 90-120 minute; Conversion fluid is transferred pH0.5-2.5 with hydrochloric acid, and the 25%-50% that presses the conversion fluid volume adds organic solvent, stirs down at 5-20 ℃, leaves standstill 20 minutes, separates; Water is transferred pH1.5-2.7 with ammoniacal liquor, adds crystal seed and growing the grain 10 minutes, continues to transfer pH2.8-3.2 with ammoniacal liquor, adds crystallizing agent, continues to transfer to iso-electric point 4.0-4.3 with ammoniacal liquor, crystallization, drying.
The said organic solvent that gets in the c step can be selected butanols, butyl ester, methyl iso-butyl ketone (MIBK), methylene dichloride or toluene for use.But preferable methyl isobutyl ketone.
Crystallizing agent can be acetone, methyl alcohol or methyl iso-butyl ketone (MIBK), preferred acetone.
Crystallizing agent is as selecting acetone, methyl alcohol for use, and its add-on is counted 40-70% by the crystal solution volume, preferred 50-60%.
Crystallizing agent is as selecting methyl iso-butyl ketone (MIBK) for use, and its add-on is counted 5-30% by the crystal solution volume, preferred 10-20%.
Innovation part of the present invention has been to reduce the crystallization number of times of prior art in preparation 6-APA process, finish brilliant this processing step of penicillin Industrial Salt that forms thereby omit, the yield losses of effectively having avoided the penicillin crystallization to be caused, make the 6-APA yield improve 7~10%, reach more than 91%, significantly reduce the 6-APA production cost, also improved labour productivity simultaneously.
Embodiment
Embodiment 1
The Penicillium notatum fermented liquid after filtration, acidifying, butyl ester concentration extraction obtain tiring and be 7-10 ten thousand unit penicillinic acid butyl ester extracting solutions, and (i.e. BA) presses the 40-100g/ BOU and add gac, stirred 20-60 minute in-20-10 ℃, filters; To remove the part foreign pigment among the BA one time, filter, in filtrate, add 9~12% solution of potassium carbonate by 1.0-1.5 liter/BOU, stir, leave standstill phase-splitting, back extraction gets the benzylpenicillin potassium aqueous solution (RB), RB is tired be controlled at 45--55 ten thousand units, guarantee to take off the ester effect; RB carries out degreasing with the velocity flow of 200-400L/h through ADS-18 (Tianjin Compositech Inc. of Nankai produces and sells), to remove butyl ester residual among the RB, wash resin column with the purified water top, the 1.5-3 that the benzylpenicillin potassium aqueous solution after the degreasing (being elutriant) volume reaches (before being degreasing) volume before the upper prop doubly, the wash-out multiple increases and increases along with RB tires.Improve the multiple of elutriant, can guarantee that benzylpenicillin potassium is fully eluted; Elutriant is placed the retort that penicillin acylase is housed; the 3%-10% that presses the elutriant cumulative volume adds borate buffer; carry out conversion reaction; stir; with the toluylic acid that produces in the ammoniacal liquor and in the conversion reaction process, make conversion reaction system pH maintain 6.5-9.0 simultaneously, the conversion reaction temperature is 25-35 ℃; reacted 90-120 minute; filter, transform filtrate and transfer pH0.5-2.5 with 20% hydrochloric acid (w/v), the 25%-50% that presses the conversion fluid volume adds butanols; stir down at 5-20 ℃; left standstill 20 minutes, and separated discarding organic phase, to remove the part oil-soluble impurities; water is transferred pH1.5-2.7 with ammoniacal liquor; add crystal seed and growing the grain 10 minutes, continue to transfer pH2.8-3.2, add crystallizing agent acetone with ammoniacal liquor; consumption is 40% of a crystal solution volume; continuation transfers to isoelectric pH 4.0-4.3 with ammoniacal liquor, and growing the grain 3-4 hour, the crystallization control terminal temperature was below 10 ℃; crystallization; filter; drying, the yield of gained 6-APA finished product reaches more than 92.3%.
Embodiment 2
The Penicillium notatum fermented liquid after filtration, acidifying, the butyl ester concentration extraction obtains tiring is 7.3 ten thousand unit penicillin butyl ester extracting solutions, and promptly a BA presses the 40g/ BOU and adds gac, stirred 20 minutes in-20-10 ℃, filters; Removing the part foreign pigment among the BA one time, filter, in filtrate, adds 9% sodium carbonate solution by 1.0 liters/BOU, phase-splitting is left standstill in stirring, separate the benzylpenicillin sodium aqueous solution (RB), it is 550,000 units that RB tires; RB carries out degreasing with the velocity flow of 400L/h through AXT-33 (having commercially available) resin column, removing butyl ester residual among the RB, washes resin column with the purified water top, makes the volume of elutriant reach 3 times of upper prop front volume; Elutriant is placed the retort that penicillin acylase is housed, add borate buffer by 3% of elutriant cumulative volume after the degreasing, stir, with the toluylic acid that produces in the ammoniacal liquor and in the conversion reaction process, make conversion reaction system pH maintain 6.5 simultaneously, the conversion reaction temperature is 25 ℃, reacted 90 minutes, filter, transform filtrate and transfer pH0.5, press 25% of conversion fluid volume and add methyl iso-butyl ketone (MIBK) with 20% hydrochloric acid (w/v); Stir down at 5-20 ℃, leave standstill, separate discarding organic phase, to remove the part oil-soluble impurities, water is transferred pH1.5-2.7 with ammoniacal liquor, adds crystal seed and growing the grain 10 minutes, continue to transfer ammoniacal liquor to transfer pH2.8-3.2, add methyl alcohol, the consumption of methyl alcohol is 60% of a crystal solution volume, continuation transfers to isoelectric pH 4.0 with ammoniacal liquor, growing the grain 3 hours, crystallization control terminal temperature filter at 8 ℃, drying, the yield of gained 6-APA finished product reaches 92.3%.
Embodiment 3
The Penicillium notatum fermented liquid after filtration, acidifying, the butyl ester concentration extraction obtains tiring is 100,000 unit penicillinic acid butyl ester extracting solutions, and promptly a BA presses the 100g/ BOU and adds gac, stirred 50 minutes in-20-10 ℃, filters; Removing the part foreign pigment among the BA one time, filter, in filtrate, adds 12% sodium carbonate solution by 1.5 liters/BOU, phase-splitting is left standstill in stirring, separate the benzylpenicillin sodium aqueous solution (RB), it is 500,000 units that RB tires; RB carries out degreasing with the velocity flow of 200L/h through the AXT-33 resin column, removing butyl ester residual among the RB, washes resin column with the purified water top, makes the volume of elutriant reach 3 times of upper prop front volume; Elutriant is placed the retort that penicillin acylase is housed, add borate buffer by 10% of degreasing elutriant cumulative volume, stir, with the toluylic acid that produces in the ammoniacal liquor and in the conversion reaction process, make conversion reaction system pH maintain 6.5 simultaneously, the conversion reaction temperature is 25 ℃, reacted 120 minutes, filter, filtrate is transferred pH0.5 with 20% hydrochloric acid (w/v), presses 25% of filtrate volume and adds butanols; Stir down at 5-20 ℃, left standstill 20 minutes, separate discarding organic phase, to remove the part oil-soluble impurities, water is transferred pH1.5-2.7 with ammoniacal liquor, adds crystal seed and growing the grain 10 minutes, continue to transfer ammoniacal liquor to transfer pH2.8-3.2, add methyl iso-butyl ketone (MIBK), consumption is 30% of a crystal solution volume, continuation transfers to isoelectric pH 4.2 with ammoniacal liquor, growing the grain 3 hours, crystallization control terminal temperature filter at 5 ℃, drying, the yield of gained 6-APA finished product reaches 94.1%.