CN103937866B - A kind of preparation method of the ampicillin of improvement - Google Patents
A kind of preparation method of the ampicillin of improvement Download PDFInfo
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Abstract
The present invention relates to pharmaceutical field, provide a kind of method preparing ampicillin and the product of the method acquisition, comprising: 1) under 0 DEG C～30 DEG C conditions, 6-APA is mixed in water in the synzyme of addition ampicillin with D-PG methyl ester or its salt, being 5.5～7.5 at pH value, temperature is react at 10 DEG C～30 DEG C；2) by step 1) products therefrom, it is adjusted to solution clarification with acid, keeping pH value is 0.5～2.0, and temperature is 10 DEG C-30 DEG C；3) by step 2) gained solution, be cooled to 12 DEG C～15 DEG C, then regulate pH to 3.00～3.50, then be cooled to 5 DEG C～6 DEG C, pH value maintain 3.50；Then adjust pH 4.9～5.0 to cool to 1 DEG C～2 DEG C maintenances, obtain ampicillin crystallization.The present invention improves yield and the quality of ampicillin product significantly, improves production efficiency, ensure that the drug safety of ampicillin product.
Description
Technical field
The preparation method that the present invention relates to a kind of ampicillin, the enzyme process being specifically related to a kind of improvement prepares the method for ampicillin.
Background technology
Ampicillin, chemistry (2S by name, 5R, 6R)-3,3-dimethyl-6-[(R)-2-amino-2-benzene ethylamino]-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-formic acid, its free acid is containing 3 molecular crystalline water, and it is a kind of conventional penicillins wide spectrum beta-lactam antibiotic.
Ampicillin bactericidal action, penetration cell wall very capable, be one of current widely used oral penicillin.The preparation of ampicillin has chemical synthesis and enzyme process to prepare method, and chemical method relative response condition is harsh, and reactions steps is complicated, and environmental pollution is bigger.And enzymatic synthesis method can substantially reduce reactions steps, reaction condition is gentle, is one of important channel producing ampicillin.
" research of ampicillin enzymatic synthesis method and medium system " (Pan Shengbin; college of science of Zhejiang University; master thesis; 20070901) synthetic method of ampicillin enzymatic under different medium system is reported; mainly describe the method synthesizing ampicillin under organic system and two systems; due to the requirement to environmental conservation, enzyme' s catalysis ampicillin in organic system medium, the problem that there is the pollution to environment；And aqueous phase system meets environmental requirement relatively, but the productivity of the enzyme' s catalysis ampicillin under research aqueous phase system recorded in this article is not significantly high, even if its productivity is also just 65.6% under optimum reaction condition, and this aqueous media adds organic solvent, the ratio of concentration of substrate is inappropriate (such as 1:2), D-PG methyl ester hydrochloride large usage quantity, it not most economical and be not best suited for big production, and facts have proved that the side chain quality more many, product of introducing is more poor, it is exactly impurity in product that side chain can not react；Prior art literature adds organic solvent to improve reaction efficiency, and reacting phase ratio in complete aqueous phase, is unfavorable for economic and environment-friendly, low toxicity, and anaphylactogen reduces；And how to improve the ampicillin productivity under pure water phase system, still need further research, improve simultaneously ampicillin product quality, ensure drug safety, be the target that further seeks for of drug research and development field equally.
Summary of the invention
For above technological deficiency, the invention provides the method that the enzyme process of a kind of improvement prepares ampicillin.Said method comprising the steps of:
1) under 0 DEG C～30 DEG C conditions, being mixed to join in the synzyme of ampicillin by 6-APA with D-PG methyl ester or its salt, be 5.5～7.5 at pH value in water, temperature is react at 10 DEG C～30 DEG C；
2) by step 1) products therefrom, it is adjusted to solution clarification with acid, maintenance pH value is 0.5-2.0, and temperature is 10 DEG C-30 DEG C；
3) by step 2) gained solution, be cooled to 12～15 DEG C, then regulate pH to 3.00～3.50, then be cooled to 5-6 DEG C, pH value maintain 3.50；Then adjusting pH is 4.9-5.0, cools to 1-2 DEG C of maintenance, obtains ampicillin crystallization.
The described step 1 of the present invention) in, as one of embodiment, described 6-APA and D-PG methyl ester or its salt with in be mixed to join the mode in the synzyme of ampicillin, include but not limited to: stir after the solid of 6-APA solid with D-PG methyl ester or its salt is mixed in water and be added directly in the synzyme of ampicillin；Or 6-APA solid first dissolved be then added in the synzyme of ampicillin, it is then slowly added into D-PG methyl ester or its salt liquid；Then pH value be 5.5～7.5, temperature be react under 10 DEG C～30 DEG C conditions；
The described step 1 of the present invention) in, as one of embodiment, the salt of wherein said D-PG methyl ester includes but not limited to hydrochlorate, it is also possible to be other pharmaceutical salts of D-PG methyl ester commonly used in the art.
The described step 1 of the present invention) in, as one of embodiment, described pH value is 5.5～7.5, and pH value range includes but not limited to 5.5～7.0,6.0～7.5,6.6～7.5 or 6.0～7.0；Same pH value can be 5.5,5.8,6.0,6.3,6.5,6.8,7.0,7.2 or 7.5；The described step 1 of the present invention) in, as one of embodiment, described pH value is 6.2～6.5.
The described step 1 of the present invention) in, as one of embodiment, 10 DEG C～30 DEG C scopes of described reaction temperature include but not limited to: 10 DEG C～25 DEG C, 10 DEG C～20 DEG C, 15 DEG C～20 DEG C, 15 DEG C～25 DEG C, 15 DEG C～30 DEG C, 20 DEG C～30 DEG C, 25 DEG C～30 DEG C；
As one of embodiment, described temperature can be 10 DEG C, 11 DEG C, 12 DEG C, 13 DEG C, 14 DEG C, 15 DEG C, 16 DEG C, 17 DEG C, 18 DEG C, 19 DEG C, 20 DEG C；21 DEG C, 22 DEG C, 23 DEG C, 24 DEG C, 25 DEG C, 26 DEG C, 27 DEG C, 28 DEG C, 29 DEG C or 30 DEG C；As one of embodiment, described reaction temperature is preferably 10 DEG C～15 DEG C
As one of embodiment of the present invention, described step 1) in, mixing and the temperature in course of reaction are preferably 10 DEG C～15 DEG C；PH value in course of reaction is 6.2～6.5；When the residual quantity of 6-APA is lower than 2mg/ml, terminate reaction.
When step 1 of the present invention) course of reaction in, in order to keep above-mentioned pH value range, selectively use include but not limited to that the ammonia spirit of 3N suitably regulates.
The described step 1 of the present invention) in, it is 1.0:1.0～1.5 as one of embodiment, described 6-APA and the weight ratio to Phenylglycine methyl ester hydrochlorate, it is preferable that 1.0:1.0；The concentration of described ampicillin synzyme is 1.0KU/L-1.5U/L.
The described step 2 of the present invention) in, as one of embodiment, described acid is selected from hydrochloric acid, dilute sulfuric acid, dust technology, phosphoric acid solution, oxalic acid or citric acid；The mass concentration of preferred acid ranges for: 10%-30%, it can be the concentration range of 12%～28%, 15%～25%, 18%～22%, the exemplarily explanation of property, such as hydrochloric acid that mass concentration is 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, dilute sulfuric acid, dust technology, phosphoric acid solution, oxalic acid, citric acid；As one of embodiment；It is preferably hydrochloric acid.
Step 2 described in the inventive method) in, as one of embodiment, being optionally added appropriate ETDA-2Na, wherein said " in right amount " can be determined by the amount of those skilled in the art's association reaction product, as being the 0.5%～2% of raw material 6-APA mass；The illustratively amount of 0.5%, 1%, 1.5% or 2%.
The described step 3 of the present invention) in, as one of embodiment, described alkali includes but not limited to sodium hydroxide, ammonia, potassium hydroxide, barium hydroxide or calcium hydroxide；Described paper mill wastewater is 10-40%；It can be 12%～38%, 12%～35%, 12%～30%, 12%～25%, 15%～35%, 15%～30%, 15%～25%, 18%～25%, 18%～35%, 18%～30%, 18%～25%；10%, the sodium hydroxide of 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40% mass concentration, ammonia, potassium hydroxide, barium hydroxide or aqua calcis；It is preferably the sodium hydroxide solution of 20% mass concentration.
As one of embodiment of the present invention, the inventive method may further comprise:
4) precipitate out after crystal, successively with water, selectively with 70%～95% methanol, ethanol or isopropanol, 80-98% the organic solvent washing of acetone, dry, to obtain final product.
In the inventive method, as one of real-time mode, described step 4) in, the weight ratio of water or organic solvent and ampicillin is 2～3:1, it is preferred to 1:1.
In inventive method, as one of real-time mode, described step 4) in, drying condition is: evacuation stirring is dry, and below vacuum 0.095MPa, temperature is 45-65 DEG C.
As one of embodiment, Phenylglycine methyl ester or its hydrochlorate, ampicillin synzyme and/or water are the raw material that this area is conventional by described 6-APA, D-of the present invention, meet the quality criteria requirements that this area is conventional, it is preferable that meet following standard, arbitrarily the originate raw material of (buying in city or preparation) or the raw material of higher standard.
As one of preferred embodiment of the present invention, the enzymatic-process preparation method of ampicillin of the present invention preferably includes following steps:
Feed in raw material and reaction:
1) under 10 DEG C～30 DEG C conditions, 6-APA is mixed in water in the synzyme of addition ampicillin with D-PG methyl ester or its hydrochlorate, ampicillin synzyme is 1.0KU/L-1.5KU/L. concentration, control pH value is 5.5-7.5, temperature is 10-30 DEG C, when the residual of 6-APA is lower than 2mg/ml, terminate reaction；Selectively regulate the pH value in course of reaction with 3mol/L ammonia
Crystallization:
2) by step 1) products therefrom, it is adjusted to solution clarification with hydrochloric acid, keeping pH value is 0.5～2.0, and temperature is 10 DEG C～30 DEG C；And it is optionally added appropriate EDTA-Na；
3) it is then passed through filter and enters crystallizer, be first cooled to 12～15 DEG C, then regulate pH with sodium hydroxide solution, under 3.00～3.50,5～6 DEG C of conditions, pH is maintained 3.50；Then adjust pH to be 4.9～5.0 with sodium hydroxide solution, cool to 1 DEG C～2 DEG C growing the grains；Wherein sodium hydroxide solution is 15-30% mass concentration；
Refining, dry:
4) solid-liquid separation is carried out after growing the grain, after separating whole mother solution, first purified water washing, then with the methanol of 70%～95% concentration, 80%～98% acetone or 70%～95% washing with alcohol is centrifugal, while stirring evacuation, make vacuum at below 0.095MPa, temperature keeps 45～65 DEG C, dry.
The present invention still further provides a kind of ampicillin product adopting the inventive method to prepare and to obtain, and the ampicillin content of described product is more than or equal to 99.5%；Always assorted content is less than or equal to for 0.20%；Single assorted content is less than or equal to 0.10%.
Relative to prior art, the inventive method considerably improves the yield of ampicillin product, and the quality of ampicillin product that the present invention obtains is higher, and always assorted content assorted, single significantly decreases.
Compared with recording prior art with document, the inventive method has carried out significant raising or improvement in following several respects: in pH value in reaction, and pH value, closer to neutrality, conveniently controls, it is to avoid the destruction of highly basic strong acid, further reduces the generation of impurity；From the raw material used: inapplicable poisonous and hazardous raw material, for instance: pivaloyl chloride, pyridine, triethylamine, the contour carcinogenic chemical raw material such as dichloromethane, simply last washing link is suitable for two classes and dissolves acetone, is high-efficiency low-toxicity to human body and link；Reaction temperature, temperature is compared the control of the more convenient big production of chemical method, because chemical method to react under-40 DEG C of conditions, the present invention reacts under 10 DEG C of-15 DEG C of conditions, more energy efficient, and more environmental protection is more easy to operation；From method for crystallising, crystallization process all carries out in aqueous phase, nonpoisonous and tasteless, health environment-friendly, is not added with any organic solvent, will not remain in crystals in crystallization after being added without any dissolving in product, making product quality better, sensitinogen is less, and commercial production is more efficiently easily-controllable；From refining aspect, in the selection of solvent, the methanol of 70%-95% concentration of the present invention, the acetone of 80%-98%, 70%-95% washing with alcohol, it is possible to reach better effect, as moisture content dries well, solvent residual is lower.
The present invention is by adopting the Accelerated stability test method described in pharmacopeia (second edition in 2010) to detect, and in the present invention, the ampicillin steady quality of products obtained therefrom in each embodiment, further increases the drug safety of ampicillin product.
Detailed description of the invention
The present invention is expanded on further the present invention by following example and experimental example, but the present invention is not limited to this.
Embodiment 1
Material prepares:
6-APA35g；D-PG methyl ester 29g；Ampicillin synthetase 1 .0KU/L；98% acetone 35ml；EDTA-2Na0.35g；10% hydrochloric acid 56ml；3mol/L ammonia 2ml；20% sodium hydrate aqueous solution 38ml；Purified water 585ml.
Preparation process:
Add purified water 350ml in the reactor, add 6-APA35g under stirring, D-PG methyl ester 29g stirs.
Being cooled to 10 DEG C, add ampicillin synthetase 1 .0KU/L, reaction whole process 3mol/L ammonia controls pH6.4～6.5, temperature 10 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 2ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 20 DEG C, after dripping 10% hydrochloric acid 56ml, ampicillin is dissolved, and pH=1.01 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C, dropping 36ml20% sodium hydrate aqueous solution regulates pH to 3.0～3.5, it is cooled to 5～6 DEG C and pH is maintained at 3.5,2ml20% sodium hydroxide solutions adjustment pH to 4.9～5.0, continue to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, in 45 DEG C after 35ml washing with acetone 1 time, dries under vacuum 0.09MP.Obtain ampicillin: 56.84g, yield: 86.9%.
Embodiment 2
Material prepares:
6-APA35g；D-PG methyl ester hydrochloride 35g；Ampicillin synthetase 1 .0KU/L；80% acetone 35ml；EDTA-2Na0.35g；15% hydrochloric acid 37ml；3mol/L ammonia 55ml；10% sodium hydrate aqueous solution 78ml；Purified water 585ml.
Preparation process:
Add purified water 350ml in the reactor, add 6-APA35g, D-PG methyl ester hydrochloride 35g under stirring, stir.
Being cooled to 11 DEG C, add ampicillin synthetase 1 .3KU/L, reaction whole process 3mol/L ammonia controls pH6.3～6.4, temperature 11 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 55ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 21 DEG C, after dripping 15% hydrochloric acid 37ml, ampicillin is dissolved, and pH=1.04 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C, pH to 3.0～3.5 is regulated with 10% sodium hydrate aqueous solution 74ml, it is cooled to 5～6 DEG C and keeps pH value 3.5, then 10% sodium hydrate aqueous solution 4ml regulates pH to 4.9～5.0, continues to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, and filter cake 35ml purified water is washed 1 time, in 50 DEG C after 35ml80% washing with acetone 1 time, dry under vacuum 0.09MP.Obtain ampicillin: 56.77g, yield: 86.8%.
Embodiment 3
Material prepares:
6-APA35g；D-PG methyl ester hydrochloride aqueous solution 88ml (concentration 400mg/ml)；Ampicillin synthetase 1 .2KU/L；70% methanol 35ml；EDTA-2Na0.35g；20% hydrochloric acid 28ml；3mol/L ammonia 46ml；15% sodium hydrate aqueous solution 51ml；Purified water 497ml.
Preparation process:
Add purified water 262ml in the reactor, add 6-APA35g, D-PG methyl ester hydrochloride aqueous solution 88ml under stirring, stir.
Being cooled to 12 DEG C, add ampicillin synthetase 1 .2KU/L, reaction whole process 3mol/L ammonia controls pH6.3～6.4, temperature 12 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 46ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 22 DEG C, after dripping 20% hydrochloric acid 28ml, ampicillin is dissolved, and pH=1.05 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, be cooled to 12～15 DEG C, then regulate pH to 3.0～3.5 with 15% sodium hydroxide solution 48ml, being cooled to 5～6 DEG C keeps pH value 3.50, then regulate pH to 4.9～5.0 with 15% sodium hydroxide solution 3ml, continue to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, after 35ml70% methanol washs 1 time, in 55 DEG C, dry under vacuum 0.09MP, obtain ampicillin: 56.26g, yield: 86.1%.
Embodiment 4
Material prepares:
6-APA35g；D-PG methyl ester 37g；Ampicillin synthetase 1 .5KU/L；95% methanol 35ml；EDTA-2Na0.35g；30% hydrochloric acid 19ml；3mol/L ammonia 10ml；25% sodium hydrate aqueous solution 28ml；Purified water 585ml.
Preparation process:
Add purified water 350ml in the reactor, add 6-APA35g, D-PG methyl ester 37g under stirring, stir.
Being cooled to 13 DEG C, add ampicillin synthetase 1 .5KU/L, reaction whole process 3mol/L ammonia controls pH6.3～6.4, temperature 13 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 10ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 23 DEG C, after dripping 30% hydrochloric acid 19ml, ampicillin is dissolved, and pH=1.06 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C, then 25% sodium hydrate aqueous solution 27ml regulates pH to 3.0～3.5, being cooled to 5～6 DEG C keeps pH 3.5, then regulates pH to 4.9～5.0 with 25% sodium hydrate aqueous solution 1ml, continues to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, and 35ml95% methanol washs after 1 time in 60 DEG C, dries under vacuum 0.09MP.Obtain ampicillin: 56.42g, yield: 86.3%.
Embodiment 5
6-APA35g；D-PG methyl ester hydrochloride 44g；Ampicillin synthetase 1 .4KU/L；95% ethanol 35ml；EDTA-2Na0.35g；15% hydrochloric acid 37ml；3mol/L ammonia 65ml；20% sodium hydrate aqueous solution 38ml；Purified water 585ml.
Preparation process:
Add purified water 350ml in the reactor, add 6-APA35g, D-PG methyl ester 37g under stirring, stir.
Being cooled to 14 DEG C, add ampicillin synthetase 1 .4KU/L, reaction whole process 3mol/L ammonia controls pH6.2～6.3, temperature 14 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 65ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 23 DEG C, after dripping 15% hydrochloric acid 37ml, ampicillin is dissolved, and pH=1.06 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C, pH to 3.0～3.5 is regulated with 20% sodium hydrate aqueous solution 37ml, it is cooled to 5～6 DEG C and pH value is maintained at 3.50, then regulate pH to 4.9～5.0 with 25% sodium hydrate aqueous solution 1ml, continue to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, in 60 DEG C after 95% washing with alcohol of 35ml 1 time, dries under vacuum 0.09MP.Obtain ampicillin: 56.59g, yield: 86.6%.
Embodiment 6
6-APA35g；D-PG methyl ester hydrochloride aqueous solution 116ml (concentration 380mg/ml)；Ampicillin synthetase 1 .5KU/L；80% ethanol 35ml；EDTA-2Na0.35g；15% hydrochloric acid 37ml；3mol/L ammonia 60ml；25% sodium hydrate aqueous solution 29ml；Purified water 469ml.
Preparation process:
Add purified water 234ml in the reactor, add 6-APA35g, D-PG methyl ester hydrochloride aqueous solution 116ml under stirring, stir.
Being cooled to 15 DEG C, add ampicillin synthetase 1 .5KU/L, reaction whole process 3mol/L ammonia controls pH6.2～6.3, temperature 15 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 60ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 24 DEG C, after dripping 15% hydrochloric acid 37ml, ampicillin is dissolved, and pH=1.05 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C and then regulates pH to 3.0～3.5 with 25% sodium hydrate aqueous solution 28ml, being cooled to 5～6 DEG C keeps pH to maintain 3.5, then pH to 4.9～5.0 is regulated with 25% sodium hydroxide solution 1ml, continuing to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, in 55 DEG C after 35ml80% washing with alcohol 1 time, dry under vacuum 0.09MP.Obtain ampicillin: 56.11g, yield: 85.8%.
Embodiment 7
Material prepares:
6-APA35g；D-PG methyl ester hydrochloride 43g；Ampicillin synthetase 1 .3KU/L；80% isopropyl acetone 35ml；EDTA-2Na0.35g；10% hydrochloric acid 55ml；3mol/L ammonia 20ml；20% sodium hydrate aqueous solution 38ml；Purified water 585ml.
Preparation process:
Add purified water 350ml in the reactor, add 6-APA35g, D-PG methyl ester 43g under stirring, stir.
Being cooled to 15 DEG C, add ampicillin synthetase 1 .3KU/L, reaction whole process 3mol/L ammonia controls pH6.2～6.3, temperature 14 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 20ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 24 DEG C, after dripping 10% hydrochloric acid 55ml, ampicillin is dissolved, and pH=1.06 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C, then pH to 3.0～3.5 is regulated with 20% sodium hydrate aqueous solution 37ml, being cooled to 5～6 DEG C keeps pH value 3.50, then regulates pH to 4.9～5.0 with 20% sodium hydrate aqueous solution 1ml, continues to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, and 35ml80% isopropyl acetone washs after 1 time in 60 DEG C, dries under vacuum 0.09MP.Obtain ampicillin: 56.34g, yield: 86.2%.
Embodiment 8
Material prepares:
6-APA35g；D-PG methyl ester hydrochloride 52.5g；Ampicillin synthetase 1 .2KU/L；90% isopropanol 35ml；EDTA-2Na0.35g；15% hydrochloric acid 37ml；3mol/L ammonia 48ml；20% sodium hydrate aqueous solution 38ml；Purified water 585ml.
Preparation process:
Add purified water 350ml in the reactor, add 6-APA35g, D-PG methyl ester hydrochloride 52.5g under stirring, stir.
It is cooled to 14 DEG C, adds ampicillin synthetase 1 .2KU/L, reaction whole process 3mol/L ammonia control ph 6.4～6.5, temperature 14 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 70ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 21 DEG C, after dripping 15% hydrochloric acid 37ml, ampicillin is dissolved, and pH=1.06 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C, then pH to 3.0～3.5 is regulated with 20% sodium hydrate aqueous solution 37ml, it is cooled to 5～6 DEG C and keeps pH value 3.50, then regulate pH to 4.9～5.0 with 20% sodium hydrate aqueous solution 1ml, continue to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, and 90% isopropyl acetone of 35ml washs after 1 time in 60 DEG C, dries under vacuum 0.09MP.Obtain ampicillin: 56.77g, yield: 86.8%.
Embodiment 9
Material prepares:
6-APA35g；D-PG methyl ester hydrochloride aqueous solution 125ml (concentration 420mg/ml)；Ampicillin synthetase 1 .4KU/L；80% methanol 35ml；EDTA-2Na0.35g；15% hydrochloric acid solution 37ml；3mol/L ammonia 72ml；20% sodium hydrate aqueous solution 38ml；Purified water 460ml.
Preparation process:
Add purified water 225ml in the reactor, add 6-APA35g, D-PG methyl ester hydrochloride aqueous solution 125ml under stirring, stir.
Being cooled to 15 DEG C, add ampicillin synthetase 1 .4KU/L, reaction whole process 3mol/L ammonia controls pH6.2～6.3, temperature 15 ± 0.5 DEG C, stopped reaction when reaction is to 6-APA residual concentration < 2mg/ml, 3mol/L ammonia volume 72ml.
Ampicillin synzyme is separated, obtains ampicillin crude product.
In beaker, putting into ampicillin crude product, add 200ml purified water, be warming up to 22 DEG C, after dripping 15% hydrochloric acid 37ml, ampicillin is dissolved, and pH=1.01 adds EDTA-2Na0.35g, filters to obtain clear liquid after stirring 10min.
Clear liquid is proceeded in crystallizer, it is cooled to 12～15 DEG C, drip 20% sodium hydrate aqueous solution 38ml and regulate pH to 3.0～3.5, it is cooled to 5～6 DEG C and keeps pH3.50, then regulate pH to 4.9～5.0 with 20% sodium hydrate aqueous solution 2ml, continue to be cooled to 1～2 DEG C of growing the grain 30min, fractional crystallization liquid, filter cake 35ml purified water is washed 1 time, and 35ml80% methanol washs after 1 time in 55 DEG C, dries under vacuum 0.09MP.Obtain ampicillin: 56.69g, yield: 86.7%.
Experimental example 1 accelerated stability test
Experimental apparatus: high performance liquid chromatograph, gas chromatograph, polymer determination instrument, Karl Fischer, polariscope etc..
The condition of efficient liquid phase: wavelength: 225nm, UV-detector, flow velocity: 1.0ml/minl, mobile phase: phosphate buffer: acetonitrile=95:5,
High performance liquid chromatograph: Shimadzu LC-20A, gas chromatograph: Shimadzu GC-2010, Karl Fischer: logical 870 types of Switzerland ten thousand, polariscope: numeral automatic polarimeter WZZ-2S
Experimental technique: at each point in time sampling, the method about accelerated stability recorded by pharmacopeia 2010 editions detects.Point take employing embodiment 6, embodiment 7, embodiment 8, sample 60g prepared by the method for embodiment 9, seal bag two-layer and seal, be positioned over temperature be 40 DEG C, the accelerated test case of humidity 75% ± 5%.
Experimental result:
The accelerated test result of embodiment 6 gained ampicillin product
(40 DEG C ± 2 DEG C, RH75% ± 5%)
The accelerated test result of embodiment 7 gained ampicillin product
The accelerated test result of embodiment 8 gained ampicillin product
The accelerated test result of embodiment 9 gained ampicillin product
Experiment conclusion: Acceleration study is after 6 months, the content of gained ampicillin of the present invention product, single assorted, always assorted and polymer etc. all significantly change, and the steady quality of enzyme process ampicillin of the present invention product is described.
The present invention not only yield aspect compared with prior art has and is greatly improved, and the content of the ampicillin product of gained, single assorted, always assorted and polymer have and significantly reduce and accelerated test all significantly changed after 6 months, the steady quality of enzyme process ampicillin of the present invention product is described.
Claims (13)
1. the method that enzyme process prepares ampicillin, described method includes:
1) under 0 DEG C～30 DEG C conditions, 6-APA is mixed in water in the synzyme of addition ampicillin with D-PG methyl ester or its hydrochlorate, it is 5.5～7.5 at pH value, temperature is react at 10 DEG C～30 DEG C, wherein 6-APA and the mass ratio to Phenylglycine methyl ester or its hydrochlorate are 1.0:1.0～1.5, and the concentration of described ampicillin synzyme is 1.0KU/L-1.5KU/L；
2) by step 1) products therefrom, it is adjusted to solution clarification with acid, keeping pH value is 0.5～2.0, and temperature is 10 DEG C～30 DEG C, adds appropriate EDTA-2Na；
3) by step 2) gained solution, it is cooled to 12～15 DEG C, then alkali regulates pH value to 3.00～3.50, then is cooled to 5 DEG C～6 DEG C, maintains pH value 3.50；Then alkali tune pH value is 4.9～5.0, cools to 1 DEG C～2 DEG C maintenances, obtains ampicillin crystallization.
2. method according to claim 1, it is characterised in that described step 1) in, 6-APA and the mass ratio to Phenylglycine methyl ester or its hydrochlorate are 1.0:1.0.
3. method according to claim 1, it is characterised in that described step 1) in, the temperature of course of reaction is 10 DEG C～15 DEG C；PH value in course of reaction is 6.2～6.5；Or when the residual quantity of 6-APA is lower than 2mg/ml, terminate reaction.
4. method according to claim 1, it is characterised in that described step 2) in temperature be 20 DEG C～24 DEG C.
5. method according to claim 1, it is characterised in that described step 2) in acid selected from hydrochloric acid, sulphuric acid, nitric acid, phosphoric acid solution, oxalic acid or citric acid.
6. method according to claim 5, it is characterised in that described step 2) in acid mass concentration range for 10%～30%.
7. method according to claim 1, it is characterised in that described step 3) in alkali selected from sodium hydroxide, ammonia, potassium hydroxide, barium hydroxide or calcium hydroxide.
8. method according to claim 7, it is characterised in that described step 3) in the mass concentration of alkali be 10～40%.
9. method according to claim 1, it is characterised in that described method also includes:
4) step 3) precipitate out after crystal, with water or water and 70%～95% methanol, ethanol or isopropanol or 80-98% the organic solvent washing of acetone, dry, to obtain final product.
10. method according to claim 9, it is characterised in that described step 4) in the weight ratio of water or organic solvent and ampicillin be 1:1.
11. method according to claim 9, it is characterised in that described step 4) in, drying condition is: evacuation stirring is dry, and below vacuum 0.095MPa, temperature is 45-65 DEG C.
12. according to the arbitrary described method of claim 1-11, it is characterised in that described method includes:
1) under 10 DEG C～30 DEG C conditions, 6-APA is mixed in water in the synzyme of addition ampicillin with D-PG methyl ester or its hydrochlorate, ampicillin synzyme is 1.0KU/L-1.5KU/L concentration, it is 5.5-7.5 at pH value, temperature reacts at being 10-30 DEG C, when the residual of 6-APA is lower than 2mg/ml, terminate reaction；Or regulate the pH value in course of reaction with 3mol/L ammonia；
2) by step 1) products therefrom hydrochloric acid be adjusted to solution clarification,
3) keeping pH value is 0.5-2.0, and temperature is 10 DEG C-30 DEG C；Being then passed through filter and enter crystallizer, be first cooled to 12～15 DEG C, then regulating pH with sodium hydroxide solution is 3.00～3.50, under 5-6 DEG C of condition, pH is maintained 3.50；Then adjust pH to be 4.9-5.0 with sodium hydroxide solution, cool to 1-2 DEG C of growing the grain；Wherein sodium hydroxide solution is 15-30% concentration；
4) solid-liquid separation is carried out after growing the grain, after separating whole mother solution, first carry out purified water washing, then with the methanol of 70%-95% concentration, the acetone of 80%-98%, or 70%-95% washing with alcohol is centrifugal, while stirring evacuation, make vacuum at below 0.095MPa, temperature keeps 45-65 DEG C, dry.
13. the ampicillin product that the arbitrary described method of claim 1-12 obtains, it is characterised in that in described product, ampicillin content is more than or equal to 99.5%；Always assorted content is less than or equal to for 0.20%；With single assorted content less than or equal to 0.10%.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008110527A1 (en) * | 2007-03-09 | 2008-09-18 | Dsm Ip Assets B.V. | Process for the preparation of beta-lactam compounds |
| CN102660621A (en) * | 2012-05-04 | 2012-09-12 | 联邦制药(内蒙古)有限公司 | Improved method for preparing amoxicillin by enzymic method |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008110527A1 (en) * | 2007-03-09 | 2008-09-18 | Dsm Ip Assets B.V. | Process for the preparation of beta-lactam compounds |
| CN102660621A (en) * | 2012-05-04 | 2012-09-12 | 联邦制药(内蒙古)有限公司 | Improved method for preparing amoxicillin by enzymic method |
Non-Patent Citations (2)
| Title |
|---|
| 氨苄西林结晶工艺研究;王金良;《河北科技大学硕士学位论文》;20110517;全文 * |
| 氨苄西林酶促合成方法与介质体系研究;潘绳彬;《中国优秀硕士学位论文全文数据库工程科技I辑》;20090115(第1期);第30页第1段，第32页第2段，第33页第1段 * |
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