阅读说明：本技术 一种盐酸去氧肾上腺素杂质标准品的制备方法 (Preparation method of phenylephrine hydrochloride impurity standard ) 是由 蔡锦镇 于 2020-07-01 设计创作，主要内容包括：本发明提供了一种盐酸去氧肾上腺素杂质标准品的制备方法,先将盐酸去氧肾上腺素原料药置于光照条件下,使其产生杂质,然后采用制备色谱法获得色谱纯目标杂质；再在酮基还原酶KRED和还原型辅酶Ⅱ NADPH存在的条件下,以硝基甲烷和间羟基苯甲醛为原料反应得到浓缩物；最后将浓缩物利用异丙醇-乙酸正丙酯-乙腈混合溶液进行重结晶,并在重结晶过程中加入碱中和的色谱纯目标杂质,成盐后获得一种盐酸去氧肾上腺素杂质标准品(杂质A),产品收率和纯度高,满足药物杂质标准品要求。(The invention provides a preparation method of phenylephrine hydrochloride impurity standard, which comprises the steps of putting phenylephrine hydrochloride raw material medicine under the illumination condition to generate impurities, and then obtaining chromatographically pure target impurities by adopting a preparation chromatography; reacting nitromethane and m-hydroxybenzaldehyde as raw materials in the presence of ketoreductase KRED and reduced coenzyme II NADPH to obtain a concentrate; and finally, recrystallizing the concentrate by using an isopropanol-n-propyl acetate-acetonitrile mixed solution, adding alkali-neutralized chromatographic pure target impurities in the recrystallization process, and salifying to obtain the phenylephrine hydrochloride impurity standard (impurity A), wherein the product yield and purity are high, and the requirements of the pharmaceutical impurity standard are met.)

1. A preparation method of phenylephrine hydrochloride impurity standard is characterized by comprising the following specific steps:

(1) placing phenylephrine hydrochloride raw material medicine under the illumination condition to generate impurities, and then preparing twice by adopting a preparation chromatography to obtain chromatographically pure target impurities;

(2) then in the presence of ketoreductase KRED and reduced coenzyme II NADPH, nitromethane and m-hydroxybenzaldehyde are used as raw materials to react to obtain light yellow oily matter, and then the light yellow oily matter is subjected to catalytic hydrogenation reaction, filtered and concentrated to obtain a concentrate;

(3) adding the concentrate into a mixture with the volume ratio of 1: 0.5-0.6: and (2) dissolving the isopropanol-n-propyl acetate-acetonitrile mixed solution in 0.08-0.1 by heating and stirring, adding alkali-neutralized chromatographic pure target impurities, standing for crystallization, filtering, drying under reduced pressure to obtain a solid, dissolving the solid in dry ethyl acetate, introducing dry hydrogen chloride gas, and salifying to obtain the phenylephrine hydrochloride impurity standard product.

2. The production method according to claim 1, wherein in the step (1), the light irradiation conditions are as follows: irradiating the substrate with light with a wavelength of 350-380 nm for 8-10 hours.

3. The method according to claim 1, wherein in the step (1), the apparatus conditions for the preparative chromatography are as follows: hanbang scientific NP2000, chromatographic column kromasil C1810 mu m 21.2 multiplied by 250mm, detection wavelength of 273nm, flow rate of 30mL/min, mobile phase volume ratio of 0.1: 45: 55: 0.2 of glacial acetic acid-methanol-acetonitrile-water.

4. The method according to claim 1, wherein in the step (1), the two-time preparation is carried out by the following specific method:

(A) the first preparation: firstly, phenylephrine raw material medicines are mixed according to the volume ratio of 1: dissolving 8-9 acetonitrile-n-propyl acetate, injecting into a chromatograph, collecting eluent corresponding to a peak with relative retention time of 1, and removing the solvent by rotary evaporation to obtain an oily substance;

(B) and (3) second preparation: and then the oily substance is mixed by using a volume ratio of 1: dissolving 8-9 acetonitrile-n-propyl acetate mixed solution, injecting the mixed solution into a chromatograph again, collecting eluent corresponding to a peak with relative retention time of 1, removing the solvent by rotary evaporation to obtain a concentrate, and then, mixing the concentrate with the mixed solution according to a volume ratio of 1: dissolving the 2-3 acetonitrile-n-propyl acetate mixed solution, standing for crystallization, and filtering to obtain the chromatographically pure target impurity.

5. The method according to claim 1, wherein the step (2) is specifically performed by:

(2-1) firstly, slowly dripping sodium methoxide into nitromethane at the temperature of 0 ℃, keeping the temperature and stirring for 30-40 minutes, adding ketoreductase KRED and reduced coenzyme II NADPH, then slowly dripping a nitromethane solution of m-hydroxybenzaldehyde, stirring for reaction, adding a saturated ammonium chloride solution to terminate the reaction, and carrying out aftertreatment to obtain a light yellow oily substance;

(2-2) then ultrasonically dispersing the light yellow oily substance into methanol, adding a palladium-carbon catalyst, carrying out catalytic hydrogenation, filtering and concentrating to obtain a concentrate.

6. The method according to claim 5, wherein the mass ratio of the sodium methoxide, nitromethane, ketoreductase KRED, reduced coenzyme II NADPH, the nitromethane solution of m-hydroxybenzaldehyde, methanol and palladium-carbon catalyst is 150: 10: 0.2: 0.005: 10: 3: 4, wherein the solution of m-hydroxybenzaldehyde in nitromethane is obtained by dissolving m-hydroxybenzaldehyde in nitromethane with 10 times of weight.

7. The method according to claim 1, wherein in the step (3), the mass to volume ratio of the concentrate to the isopropanol-n-propyl acetate-acetonitrile mixed solution to the base-neutralized chromatographically pure target impurity is about 1 g: 6-7 mL: 0.03-0.05 g.

8. The preparation method according to claim 1, wherein in the step (3), the temperature for heating, stirring and dissolving is 80-85 ℃, and the standing crystallization time is 6-8 hours.

9. The preparation method according to claim 1, wherein in the step (3), the alkali-neutralized chromatographic pure target impurity is obtained by adding a sodium carbonate aqueous solution with a mass concentration of 20-30% dropwise to the chromatographic pure target impurity until the chromatographic pure target impurity is neutral, and washing the chromatographic pure target impurity with high-purity water.

Technical Field

The invention relates to the technical field of pharmacy, in particular to a preparation method of a phenylephrine hydrochloride impurity standard substance.

Background

Phenylephrine hydrochloride is used for preventing and treating hypotension caused by spinal anesthesia, general anesthesia, chlorpromazine and the like, and is also used for supraventricular tachycardia, mydriasis examination and the like. Phenylephrine hydrochloride is an alpha-adrenergic receptor agonist, a sympathomimetic amine drug that acts directly on the receptor, but sometimes acts indirectly by promoting the release of phenylephrine from the site of storage, and has a similar effect to phenylephrine, but is weaker and longer-lasting, and less toxic. The traditional Chinese medicine composition is generally used in the aspects of toxicity and anaphylactic shock in infection, supraventricular cardiac acceleration and the like in clinic, has good effectiveness and safety, and has very good clinical application prospect. The structural formula of phenylephrine hydrochloride is as follows:

。

in order to ensure the safety of the medicine and reduce adverse reactions, the impurities of the medicine need to be effectively detected and controlled. The impurities found in the quality research of the phenylephrine hydrochloride medicament comprise impurity A, impurity C, impurity D, impurity E and the like, wherein the impurity A is norphenylephrine hydrochloride, the impurity is degradation impurity which is easily generated in the synthesis and storage processes of the phenylephrine hydrochloride raw material and the preparation thereof, and the structural formula is as follows:

。

patent CN100361962C discloses a method for preparing norphenylephrine hydrochloride, which is obtained by using m-hydroxybenzaldehyde as raw material, sodium methoxide or potassium hydroxide as alkali, and performing addition reaction with nitromethane, then performing catalytic hydrogenation reaction to reduce nitro, and finally performing hydrochlorination. In performing drug quality testing, high purity impurities are required as controls for controlling the quality of drugs that may contain the impurities. The norphenylephrine hydrochloride obtained by the method disclosed in the patent has low purity, cannot meet the requirements of pharmaceutical impurity standards, and has low overall yield.

Disclosure of Invention

The invention aims to provide a preparation method of phenylephrine hydrochloride impurity standard, which improves the product yield and purity and meets the requirements of pharmaceutical impurity standard.

In order to achieve the purpose, the invention is realized by the following scheme:

a preparation method of phenylephrine hydrochloride impurity standard comprises the following specific steps:

(1) placing phenylephrine hydrochloride raw material medicine under the illumination condition to generate impurities, and then preparing twice by adopting a preparation chromatography to obtain chromatographically pure target impurities;

(2) then in the presence of ketoreductase KRED and reduced coenzyme II NADPH, nitromethane and m-hydroxybenzaldehyde are used as raw materials to react to obtain light yellow oily matter, and then the light yellow oily matter is subjected to catalytic hydrogenation reaction, filtered and concentrated to obtain a concentrate;

(3) adding the concentrate into a mixture with the volume ratio of 1: 0.5-0.6: and (2) dissolving the isopropanol-n-propyl acetate-acetonitrile mixed solution in 0.08-0.1 by heating and stirring, adding alkali-neutralized chromatographic pure target impurities, standing for crystallization, filtering, drying under reduced pressure to obtain a solid, dissolving the solid in dry ethyl acetate, introducing dry hydrogen chloride gas, and salifying to obtain the phenylephrine hydrochloride impurity standard product.

Preferably, in step (1), the illumination conditions are as follows: irradiating the substrate with light with a wavelength of 350-380 nm for 8-10 hours.

Preferably, in step (1), the apparatus conditions for preparative chromatography are as follows: hanbang scientific NP2000, chromatographic column kromasil C1810 mu m 21.2 multiplied by 250mm, detection wavelength of 273nm, flow rate of 30mL/min, mobile phase volume ratio of 0.1: 45: 55: 0.2 of glacial acetic acid-methanol-acetonitrile-water.

Preferably, in step (1), the specific method for the two preparations is as follows:

(A) the first preparation: firstly, phenylephrine raw material medicines are mixed according to the volume ratio of 1: dissolving 8-9 acetonitrile-n-propyl acetate, injecting into a chromatograph, collecting eluent corresponding to a peak with relative retention time of 1, and removing the solvent by rotary evaporation to obtain an oily substance;

(B) and (3) second preparation: and then the oily substance is mixed by using a volume ratio of 1: dissolving 8-9 acetonitrile-n-propyl acetate mixed solution, injecting the mixed solution into a chromatograph again, collecting eluent corresponding to a peak with relative retention time of 1, removing the solvent by rotary evaporation to obtain a concentrate, and then, mixing the concentrate with the mixed solution according to a volume ratio of 1: dissolving the 2-3 acetonitrile-n-propyl acetate mixed solution, standing for crystallization, and filtering to obtain the chromatographically pure target impurity.

Further preferably, in the step (B), the standing and crystallization time is 5 to 6 hours.

Preferably, the specific method of step (2) is as follows:

(2-1) firstly, slowly dripping sodium methoxide into nitromethane at the temperature of 0 ℃, keeping the temperature and stirring for 30-40 minutes, adding ketoreductase KRED and reduced coenzyme II NADPH, then slowly dripping a nitromethane solution of m-hydroxybenzaldehyde, stirring for reaction, adding a saturated ammonium chloride solution to terminate the reaction, and carrying out aftertreatment to obtain a light yellow oily substance;

(2-2) then ultrasonically dispersing the light yellow oily substance into methanol, adding a palladium-carbon catalyst, carrying out catalytic hydrogenation, filtering and concentrating to obtain a concentrate.

More preferably, the mass ratio of the sodium methoxide, nitromethane, ketoreductase KRED, reduced coenzyme II NADPH, the nitromethane solution of m-hydroxybenzaldehyde, methanol and palladium carbon catalyst is 150: 10: 0.2: 0.005: 10: 3: 4, wherein the solution of m-hydroxybenzaldehyde in nitromethane is obtained by dissolving m-hydroxybenzaldehyde in nitromethane with 10 times of weight.

Preferably, the sodium methoxide is obtained by adding metal sodium into methanol at 0 ℃ and stirring until the metal sodium is dissolved, wherein the mass volume ratio of the metal sodium to the methanol is 150 mg: 1 mL.

Further preferably, the process conditions of the stirring reaction are as follows: stirring and reacting for 10-12 hours at 25 ℃.

Further preferably, in the step (2-1), the post-treatment is performed by the following specific method: the reaction mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated.

Further preferably, in the step (2-2), the supported amount of the palladium on carbon catalyst is 10%.

Further preferably, in the step (2-2), the process conditions of the catalytic hydrogenation are as follows: stirring the mixture for 10 to 20 hours at 25 ℃ and under 1 standard atmospheric pressure of hydrogen.

Further preferably, in the step (2-2), the filtration is carried out using a diatomaceous earth.

Preferably, in the step (3), the mass volume ratio of the concentrate to the isopropanol-n-propyl acetate-acetonitrile mixed solution and the alkali-neutralized chromatographically pure target impurity is 1 g: 6-7 mL: 0.03-0.05 g.

Preferably, in the step (3), the temperature for heating, stirring and dissolving is 80-85 ℃, and the standing and crystallization time is 6-8 hours.

Preferably, in the step (3), the alkali-neutralized chromatographically pure target impurity is obtained by dropwise adding a sodium carbonate aqueous solution with the mass concentration of 20-30% to the chromatographically pure target impurity until the target impurity is neutral, and washing the target impurity with high-purity water.

The invention has the beneficial effects that:

the method comprises the steps of firstly placing phenylephrine hydrochloride raw material medicine under the illumination condition to generate impurities, and then obtaining chromatographically pure target impurities by adopting a preparation chromatography method; reacting nitromethane and m-hydroxybenzaldehyde as raw materials in the presence of ketoreductase KRED and reduced coenzyme II NADPH to obtain a concentrate; and finally, recrystallizing the concentrate by using an isopropanol-n-propyl acetate-acetonitrile mixed solution, adding alkali-neutralized chromatographic pure target impurities in the recrystallization process, and salifying to obtain the phenylephrine hydrochloride impurity standard (impurity A), wherein the product yield and purity are high, and the requirements of the pharmaceutical impurity standard are met.

According to the invention, the chromatographic pure target impurities are obtained by adopting the preparative chromatography, and a small amount of the chromatographic pure target impurities are added in the recrystallization process to be used as crystal nuclei, so that the phenomenon that other impurities become crystal nuclei and are wrapped in the crystal nuclei to influence the product purity is avoided. The applicant selects a solvent, uses a mixed solution of isopropanol-n-propyl acetate-acetonitrile with a specific ratio as a crystallization solvent, and greatly improves the product purity.

In a reaction system taking nitromethane and m-hydroxybenzaldehyde as raw materials, ketoreductase KRED and reduced coenzyme II NADPH are added to jointly play a role in antioxidation, so that the phenomenon that other impurities are generated by oxidation in the reaction process to influence the purity of a product is avoided. The light yellow oily substance obtained by the reaction does not need column chromatography separation, and enters the next reaction after simple post-treatment, so that the product loss is greatly reduced, the product yield is improved, and meanwhile, the product purity is ensured by combining with the special treatment of subsequent recrystallization.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The invention relates to a phenylephrine hydrochloride raw material medicine which is purchased from Shangke biological medicine (Shanghai) Co.