Gastrodin injection as well as preparation method and application thereof

Abstract

The invention discloses a preparation method of gastrodin injection and relates to the technical field of pharmaceutical preparations, gastrodin is synthesized by starting with a synthesis process of gastrodin, taking 4-formylphenyl-2, 3, 4, 6-tetra-O-acetyl-beta-D-glucopyranoside as an initial raw material, and performing reduction reaction and deacetylation reaction. The yield and the purity of the gastrodin are improved, and efficient synthesis of the gastrodin is realized; gastrodin is prepared into the injection, and the injection directly enters blood circulation after being injected, takes effect quickly and is suitable for emergency treatment of acute symptoms.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical preparations, in particular to a preparation method of gastrodin injection. BACKGROUND

[0002] Gastrodin is a glycoside compound with the chemical name of 4-hydroxymethylphenyl-β-D-glucopyranoside, a molecular formula of C 13 H 18 O7, a molecular weight of 286.278, and a CAS number of 62499-27-8. Gastrodin is mainly used to improve nervous system dysfunction and related symptoms, has the effects of calming the nerves, promoting sleep, and relieving pain, and can help regulate the excitation and inhibition functions of the cerebral cortex and restore the balance between the two. Common dosage forms of gastrodin include tablets, capsules, and injections. Tablets are convenient to take, accurate in dosage, and suitable for daily use. Capsules can reduce gastrointestinal irritation by wrapping the drug ingredients, and are suitable for people who need to take them for a long time. Injections have a quick onset and are suitable for emergency treatment or scenarios that require a quick effect.

[0003] Since gastrodin is easily soluble in water, it is suitable for being made into an injection. The main component of gastrodin injection is gastrodin, and the auxiliary materials usually include water for injection, pH adjustors, and isotonicity adjustors (such as sodium chloride and glucose). The role of the pH adjustor is to adjust the pH of the gastrodin injection to an appropriate value, ensuring the stability of gastrodin, preventing gastrodin from degrading or losing effectiveness, and also reducing the irritation of gastrodin to blood vessels or tissues. The role of the isotonicity adjustor is to adjust the osmotic pressure of the gastrodin injection to be close to that of human plasma, so as to avoid pain or tissue damage caused by the difference in osmotic pressure during injection.

[0004] The traditional source of gastrodin is extracted from the dried rhizomes of the orchid plant gastrodia elata, and it can also be artificially synthesized by chemical or biological means. Although plant extraction method has a wide source, it has a long extraction period, high cost, low yield and purity, and the yield and quality of gastrodia elata grown in artificial environment differ greatly. Chemical synthesis method has the advantages of large yield, high purity, and easy scale production, but the cost, process environmental friendliness, and control limits of impurities and harmful substances need to be considered, otherwise it cannot be industrialized and popularized. As a new method of obtaining gastrodin, biological synthesis method shows good application prospect, but there are still many challenges to achieve efficient synthesis and utilization. SUMMARY

[0005] The technical problem to be solved by the present application is to provide a preparation method of gastrodin injection, starting from the synthesis process of gastrodin, improving the yield and purity of gastrodin, controlling the types and contents of impurities, and thereby improving the stability, safety and efficacy of gastrodin injection.

[0006] The technical problem to be solved by the present invention is achieved by the following technical solution: This invention provides a method for preparing gastrodin injection, comprising the following steps: (1) 4-Formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside (compound 1) was reduced to 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside (compound 2). (2) Gastrodin was obtained by deacetylation of 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside; (3) Dissolve gastrodin completely in water for injection, then add pH adjuster and isotonic adjuster, mix evenly, filter, fill, and sterilize to obtain gastrodin injection solution.

[0007] The synthetic route of gastrodin is as follows:

[0008] Furthermore, the reduction reaction uses sodium borohydride or potassium borohydride as the reducing agent, preferably sodium borohydride.

[0009] Furthermore, the amount of reducing agent used is 1 to 2 times the molar amount of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside. Theoretically, 1 mole of sodium borohydride can reduce 4 moles of aldehyde groups. To ensure complete reaction, this invention uses excess sodium borohydride, and the reaction is quenched with water during post-treatment.

[0010] Furthermore, the reduction reaction uses barium titanate as a catalyst. To improve reduction efficiency and selectivity, this invention uses barium titanate or ruthenium dicerocene as the catalyst for the reduction reaction.

[0011] Furthermore, the reduction reaction uses ruthenium dicene as a catalyst. Compared to barium titanate, ruthenium dicene exhibits better catalytic activity for the reduction reaction.

[0012] Furthermore, the amount of the catalyst used is 3 to 5% of the mass of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0013] Furthermore, the deacetylation reaction is carried out in a sodium methoxide / methanol or sodium ethoxide / ethanol system. The sodium ethoxide acts as a catalyst, converting the ester group to a hydroxyl group via deacetylation.

[0014] Furthermore, the pH adjuster includes, but is not limited to, one or more of the following: acetic acid, hydrochloric acid, phosphoric acid, malic acid, citric acid, tartaric acid, oxalic acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium citrate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate. The function of the pH adjuster is to bring the pH value of the injection solution closer to the internal environment of the human body, reduce the irritation of the injection solution to the blood vessel walls or tissues, prevent drug degradation reactions, and prolong the drug's shelf life.

[0015] Furthermore, the isotonic adjuster includes, but is not limited to, one or more of sodium chloride, glucose, sucrose, lactose, fructose, mannitol, and sorbitol. The function of the isotonic adjuster is to adjust the osmotic pressure of the injection solution to be close to that of human blood plasma, thus avoiding pain, hemolysis, or tissue damage during injection.

[0016] Furthermore, the pH value of the gastrodin injection solution is 5.0~7.0.

[0017] Furthermore, the osmotic pressure of the gastrodin injection solution is 280~310 mOsm / kg.

[0018] Furthermore, the filtration is membrane filtration. Membrane filtration can remove impurities such as particles, bacteria, and pyrogens from the injection solution, improving the safety and efficacy of the injection solution.

[0019] Furthermore, the pore size of the filter membrane is 0.22 μm or 0.45 μm. The 0.45 μm filter membrane can filter out some bacteria as well as larger particles and impurities; the 0.22 μm filter membrane can effectively intercept most bacteria, particles and impurities.

[0020] Furthermore, the sterilization includes, but is not limited to, one of autoclaving, dry heat sterilization, and radiation sterilization. The purpose of sterilization is to ensure that the injection solution is in a sterile state and to prevent microbial contamination.

[0021] The beneficial effects of this invention are as follows: This invention starts with the synthesis process of gastrodin, using 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside as the starting material, and synthesizes gastrodin through reduction and deacetylation reactions, thereby improving the yield and purity of gastrodin and achieving efficient synthesis of gastrodin; and the gastrodin is made into an injection solution, which directly enters the blood circulation after injection, has a rapid onset of action, and is suitable for emergency treatment of acute symptoms. Detailed Implementation

[0022] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below with reference to specific embodiments.

[0023] Example 1 (1) Add 20 mmol of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and 20 mmol of sodium borohydride to 200 mL of methanol, stir to dissolve, then add barium titanate accounting for 5% of the mass of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside, and reflux for 5 h; filter (50 μm filter paper), take the filtrate, add 100 mL of water to quench the reaction; filter (50 μm filter paper), take the filtrate, concentrate under reduced pressure to obtain 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0024] (2) Add 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside prepared in step (1) and 80 mmol sodium methoxide to 250 mL of methanol, and reflux for 5 h; adjust the pH of the solution to 7.0 with acetic acid, filter (using 50 μm filter paper), collect the filtrate, concentrate under reduced pressure, and recrystallize the obtained solid with methanol-ethyl acetate (volume ratio 1:8) to obtain gastrodin. The yield was 98.24%, and the purity was 99.71%.

[0025] (3) Dissolve the gastrodin prepared in step (2) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0026] Example 2 (1) Add 20 mmol of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and 20 mmol of sodium borohydride to 200 mL of methanol, stir to dissolve, then add barium titanate accounting for 4% of the mass of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside, and reflux for 3 h; filter (50 μm filter paper), take the filtrate, add 100 mL of water to quench the reaction; filter (50 μm filter paper), take the filtrate, concentrate under reduced pressure to obtain 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0027] (2) Add 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside prepared in step (1) and 100 mmol sodium methoxide to 250 mL of methanol, and reflux for 5 h; adjust the pH of the solution to 7.0 with acetic acid, filter (using 50 μm filter paper), collect the filtrate, concentrate under reduced pressure, and recrystallize the obtained solid with methanol-ethyl acetate (volume ratio 1:5) to obtain gastrodin. The yield was 97.53%, and the purity was 99.68%.

[0028] (3) Dissolve the gastrodin prepared in step (2) completely with water for injection, then add citric acid and sodium chloride, mix evenly, adjust the pH value to 5.5, control the osmotic pressure at 290 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0029] Example 3 (1) Add 20 mmol of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and 30 mmol of sodium borohydride to 150 mL of methanol, stir to dissolve, then add barium titanate accounting for 3% of the mass of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside, and reflux for 4 h; filter (50 μm filter paper), take the filtrate, add 100 mL of water to quench the reaction; filter (50 μm filter paper), take the filtrate, concentrate under reduced pressure to obtain 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0030] (2) Add 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside prepared in step (1) and 120 mmol sodium methoxide to 250 mL of methanol, and reflux for 5 h; adjust the pH of the solution to 7.0 with acetic acid, filter (using 50 μm filter paper), collect the filtrate, concentrate under reduced pressure, and recrystallize the obtained solid with methanol-ethyl acetate (volume ratio 1:10) to obtain gastrodin. The yield was 97.18%, and the purity was 99.54%.

[0031] (3) Dissolve the gastrodin prepared in step (2) completely with water for injection, then add tartaric acid and glucose, mix evenly, adjust the pH value to 7.0, control the osmotic pressure at 310 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0032] Example 4 (1) Add 20 mmol of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and 40 mmol of sodium borohydride to 150 mL of methanol, stir to dissolve, then add barium titanate accounting for 5% of the mass of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside, and reflux for 3 h; filter (50 μm filter paper), take the filtrate, add 100 mL of water to quench the reaction; filter (50 μm filter paper), take the filtrate, concentrate under reduced pressure to obtain 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0033] (2) Add 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside prepared in step (1) and 100 mmol sodium methoxide to 250 mL of methanol, and reflux for 4 h; adjust the pH of the solution to 7.0 with acetic acid, filter (using 50 μm filter paper), collect the filtrate, concentrate under reduced pressure, and recrystallize the obtained solid with methanol-ethyl acetate (volume ratio 1:8) to obtain gastrodin. The yield was 98.07%, and the purity was 99.62%.

[0034] (3) Dissolve the gastrodin prepared in step (2) completely with water for injection, then add citric acid and sodium chloride, mix evenly, adjust the pH value to 6.5, control the osmotic pressure at 280 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0035] Example 5 Gastrodin injection was prepared according to the method in Example 1, except that barium titanate was replaced with ruthenium dicerocene during the synthesis of gastrodin.

[0036] (1) Add 20 mmol of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and 20 mmol of sodium borohydride to 200 mL of methanol, stir to dissolve, then add 5% ruthenium thiocene by mass of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and reflux for 5 h; filter (50 μm filter paper), take the filtrate, add 100 mL of water to quench the reaction; filter (50 μm filter paper), take the filtrate, concentrate under reduced pressure to obtain 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0037] (2) Add 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside prepared in step (1) and 80 mmol sodium methoxide to 250 mL of methanol, and reflux for 5 h; adjust the pH of the solution to 7.0 with acetic acid, filter (using 50 μm filter paper), collect the filtrate, concentrate under reduced pressure, and recrystallize the obtained solid with methanol-ethyl acetate (volume ratio 1:8) to obtain gastrodin. The yield was 98.96%, and the purity was 99.82%.

[0038] (3) Dissolve the gastrodin prepared in step (2) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0039] Example 6 Gastrodin injection was prepared according to the method in Example 2, except that barium titanate was replaced with ruthenium dicerocene during the synthesis of gastrodin.

[0040] (1) Add 20 mmol of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and 20 mmol of sodium borohydride to 200 mL of methanol, stir to dissolve, then add 4% (by mass) of ruthenium thiocene of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside, and reflux for 3 h; filter (using 50 μm filter paper), take the filtrate, add 100 mL of water to quench the reaction; filter (using 50 μm filter paper), take the filtrate, concentrate under reduced pressure to obtain 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0041] (2) Add 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside prepared in step (1) and 100 mmol sodium methoxide to 250 mL of methanol, and reflux for 5 h; adjust the pH of the solution to 7.0 with acetic acid, filter (using 50 μm filter paper), collect the filtrate, concentrate under reduced pressure, and recrystallize the obtained solid with methanol-ethyl acetate (volume ratio 1:8) to obtain gastrodin. The yield was 98.92%, and the purity was 99.85%.

[0042] (3) Dissolve the gastrodin prepared in step (2) completely with water for injection, then add citric acid and sodium chloride, mix evenly, adjust the pH value to 5.5, control the osmotic pressure at 290 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0043] Comparative Example 1 Gastrodin injection was prepared according to the method in Example 1, except that no catalyst was added during the synthesis of gastrodin.

[0044] (1) Add 20 mmol of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside and 20 mmol of sodium borohydride to 200 mL of methanol, stir to dissolve and reflux for 5 h; filter (50 μm filter paper), take the filtrate and add 100 mL of water to quench the reaction; filter (50 μm filter paper), take the filtrate and concentrate under reduced pressure to obtain 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

[0045] (2) Add 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside prepared in step (1) and 80 mmol sodium methoxide to 250 mL of methanol, and reflux for 5 h; adjust the pH of the solution to 7.0 with acetic acid, filter (using 50 μm filter paper), collect the filtrate, concentrate under reduced pressure, and recrystallize the obtained solid with methanol-ethyl acetate (volume ratio 1:8) to obtain gastrodin. The yield was 90.36%, and the purity was 98.92%.

[0046] (3) Dissolve the gastrodin prepared in step (2) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100mg / mL.

[0047] As can be seen from Examples 1-6 and Comparative Example 1 above, the present invention uses barium titanate and ruthenium dicerocene as catalysts for the reduction reaction, which can significantly improve the yield of gastrodin by increasing the yield of 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside. To simplify the operation, the present invention does not separate and purify 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside, but directly introduces it into the deacetylation reaction.

[0048] This invention also provides a method for preparing gastrodin injection, comprising the following steps: (1) After slicing Gastrodia elata, soak it in a solvent, reflux extract it, filter it, take the filtrate, concentrate it under reduced pressure, and separate and purify it by column chromatography to obtain gastrodin. (2) Dissolve gastrodin completely in water for injection, then add pH adjuster and isotonic adjuster, mix evenly, filter, fill, and sterilize to obtain gastrodin injection solution.

[0049] Further, the solvent is an ethanol-water-N-methylpyrrolidone mixed solution, wherein the volume ratio of ethanol to water and N-methylpyrrolidone is (55~65): (30~35): (5~10).

[0050] Furthermore, the stationary phase for column chromatography separation and purification is a polar macroporous adsorption resin, and the mobile phase is a mixed solvent of ethanol and ethyl acetate, wherein the volume ratio of ethanol to ethyl acetate is (3~5):1.

[0051] Furthermore, the pH adjuster includes, but is not limited to, one or more of the following: acetic acid, hydrochloric acid, phosphoric acid, malic acid, citric acid, tartaric acid, oxalic acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium citrate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate. The function of the pH adjuster is to bring the pH value of the injection solution closer to the internal environment of the human body, reduce the irritation of the injection solution to the blood vessel walls or tissues, prevent drug degradation reactions, and prolong the drug's shelf life.

[0052] Furthermore, the isotonic adjuster includes, but is not limited to, one or more of sodium chloride, glucose, sucrose, lactose, fructose, mannitol, and sorbitol. The function of the isotonic adjuster is to adjust the osmotic pressure of the injection solution to be close to that of human blood plasma, thus avoiding pain, hemolysis, or tissue damage during injection.

[0053] Furthermore, the pH value of the gastrodin injection solution is 5.0~7.0.

[0054] Furthermore, the osmotic pressure of the gastrodin injection solution is 280~310 mOsm / kg.

[0055] Furthermore, the filtration is membrane filtration. Membrane filtration can remove impurities such as particles, bacteria, and pyrogens from the injection solution, improving the safety and efficacy of the injection solution.

[0056] Furthermore, the pore size of the filter membrane is 0.22 μm or 0.45 μm. The 0.45 μm filter membrane can filter out some bacteria as well as larger particles and impurities; the 0.22 μm filter membrane can effectively intercept most bacteria, particles and impurities.

[0057] Furthermore, the sterilization includes, but is not limited to, one of autoclaving, dry heat sterilization, and radiation sterilization. The purpose of sterilization is to ensure that the injection solution is in a sterile state and to prevent microbial contamination.

[0058] Example 7 (1) Gastrodia elata slices were soaked in a mixed solution of ethanol-water-N-methylpyrrolidone (volume ratio of ethanol to water and N-methylpyrrolidone was 55:35:10) for 12 h at a solid-liquid ratio of 1:5. The mixture was then refluxed for 3 h, filtered (using 50 μm filter paper), and the filtrate was concentrated under reduced pressure. The filtrate was then purified by column chromatography using LSA-10 macroporous adsorption resin as the stationary phase and a mixed solvent of ethanol and ethyl acetate (volume ratio of ethanol to ethyl acetate was 5:1) to obtain gastrodin. The extraction rate was 0.54%, and the purity was 98.4%.

[0059] (2) Dissolve the gastrodin prepared in step (1) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0060] Example 8 (1) Gastrodia elata slices were soaked in a mixed solution of ethanol-water-N-methylpyrrolidone (volume ratio of ethanol to water and N-methylpyrrolidone was 60:30:10) for 12 h at a material-to-liquid ratio of 1:6. The mixture was then refluxed for 3 h, filtered (using 50 μm filter paper), and the filtrate was concentrated under reduced pressure. The filtrate was then purified by column chromatography using NKA-9 macroporous adsorption resin as the stationary phase and a mixed solvent of ethanol and ethyl acetate (volume ratio of ethanol to ethyl acetate was 3:1) to obtain gastrodin. The extraction rate was 0.59%, and the purity was 98.7%.

[0061] (2) Dissolve the gastrodin prepared in step (1) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0062] Comparative Example 2 Gastrodin injection was prepared according to the method in Example 8, except that N-methylpyrrolidone was not added during the extraction of gastrodin.

[0063] (1) Gastrodia elata slices were soaked in an ethanol-water mixture (ethanol to water volume ratio of 70:30) for 12 h at a material-to-liquid ratio of 1:6. The mixture was then refluxed for 3 h, filtered (using 50 μm filter paper), and the filtrate was collected, concentrated under reduced pressure, and purified by column chromatography. The stationary phase was NKA-9 macroporous adsorption resin, and the mobile phase was an ethanol-ethyl acetate mixture (ethanol to ethyl acetate volume ratio of 3:1). Gastrodin was obtained. The extraction rate was 0.42%, and the purity was 98.1%.

[0064] (2) Dissolve the gastrodin prepared in step (1) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0065] As can be seen from Examples 7, 8 and Comparative Example 2, the present invention uses an ethanol-water-N-methylpyrrolidone mixed solution as the soaking solvent and extraction solvent for Gastrodia elata, wherein the addition of a small amount of N-methylpyrrolidone is beneficial to improving the extraction rate of gastrodin.

[0066] The present invention also provides a method for preparing polar macroporous adsorption resin by modifying nonpolar macroporous adsorption resin, and applying the obtained polar macroporous adsorption resin to the separation and purification of gastrodin, with the aim of further improving the yield and purity of gastrodin.

[0067] The preparation method of the polar macroporous adsorption resin is as follows: Non-polar macroporous adsorption resin is swollen in toluene for 12-24 h, then chloromethyl ether and anhydrous zinc chloride are added, and the mixture is reacted at 50-60℃ for 12-24 h. After filtration, the resin is washed successively with ethanol and deionized water until no precipitate is formed after adding silver nitrate solution to the washing liquid. Subsequently, N,N-dimethylformamide, glutamine, and potassium carbonate are added, and the mixture is reacted at 70-80℃ for 8-12 h. After filtration, the resin is washed successively with ethanol and deionized water until the absorbance of the washing liquid is less than 0.005, and then dried to obtain the polar macroporous adsorption resin. The mass ratio of the non-polar macroporous adsorption resin, chloromethyl ether, anhydrous zinc chloride, glutamine, and potassium carbonate is 10 : (10-20) : (1-5) : (5-15) : (3-10).

[0068] Example 9 Gastrodin injection was prepared according to the method in Example 8, except that the NKA-9 macroporous adsorption resin was replaced with modified D-101 macroporous adsorption resin during the extraction of gastrodin.

[0069] Preparation of modified D-101 macroporous adsorption resin: 10 g of D-101 macroporous adsorption resin was swollen in 50 mL of toluene for 12 h. Then, 10 g of chloromethyl ether and 3 g of anhydrous zinc chloride were added, and the mixture was reacted at 50 °C for 12 h. The mixture was filtered (using 50 μm filter paper) and washed successively with ethanol and deionized water until no precipitate was formed after adding silver nitrate solution to the washing solution. Subsequently, 50 mL of N,N-dimethylformamide, 15 g of glutamine, and 10 g of potassium carbonate were added, and the mixture was reacted at 80 °C for 8 h. The mixture was filtered (using 50 μm filter paper) and washed successively with ethanol and deionized water until the absorbance of the washing solution was less than 0.005. The mixture was dried at 70 °C for 12 h to obtain the modified D-101 macroporous adsorption resin.

[0070] Preparation of Gastrodin Injection: (1) Gastrodia elata slices were soaked in a mixed solution of ethanol-water-N-methylpyrrolidone (volume ratio of ethanol to water and N-methylpyrrolidone was 60:30:10) for 12 h at a material-to-liquid ratio of 1:6. The mixture was then refluxed for 3 h, filtered (using 50 μm filter paper), and the filtrate was collected, concentrated under reduced pressure, and purified by column chromatography. The stationary phase was modified D-101 macroporous adsorption resin, and the mobile phase was a mixed solvent of ethanol and ethyl acetate (volume ratio of ethanol to ethyl acetate was 3:1). Gastrodin was obtained. The extraction rate was 0.64%, and the purity was 99.5%.

[0071] (2) Dissolve the gastrodin prepared in step (1) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0072] Example 10 Gastrodin injection was prepared according to the method in Example 8, except that the NKA-9 macroporous adsorption resin was replaced with modified D-101 macroporous adsorption resin during the extraction of gastrodin.

[0073] Preparation of modified D-101 macroporous adsorption resin: 10 g of D-101 macroporous adsorption resin was swollen in 50 mL of toluene for 24 h. Then, 20 g of chloromethyl ether and 4 g of anhydrous zinc chloride were added, and the mixture was reacted at 60 °C for 12 h. The mixture was filtered (using 50 μm filter paper) and washed successively with ethanol and deionized water until no precipitate was formed after adding silver nitrate solution to the washing solution. Subsequently, 50 mL of N,N-dimethylformamide, 10 g of glutamine, and 6 g of potassium carbonate were added, and the mixture was reacted at 70 °C for 12 h. The mixture was filtered (using 50 μm filter paper) and washed successively with ethanol and deionized water until the absorbance of the washing solution was less than 0.005. The mixture was dried at 70 °C for 12 h to obtain the modified D-101 macroporous adsorption resin.

[0074] Preparation of Gastrodin Injection: (1) Gastrodia elata slices were soaked in a mixed solution of ethanol-water-N-methylpyrrolidone (volume ratio of ethanol to water and N-methylpyrrolidone was 60:30:10) for 12 h at a material-to-liquid ratio of 1:6. The mixture was then refluxed for 3 h, filtered (using 50 μm filter paper), and the filtrate was collected, concentrated under reduced pressure, and purified by column chromatography. The stationary phase was modified D-101 macroporous adsorption resin, and the mobile phase was a mixed solvent of ethanol and ethyl acetate (volume ratio of ethanol to ethyl acetate was 3:1). Gastrodin was obtained. The extraction rate was 0.68%, and the purity was 99.4%.

[0075] (2) Dissolve the gastrodin prepared in step (1) completely with water for injection, then add malic acid and sodium chloride, mix evenly, adjust the pH value to 6.0, control the osmotic pressure at 300 mOsm / kg, filter (0.22μm filter membrane), fill, and autoclave (121℃ / 20 min) to obtain gastrodin injection solution with a content of 100 mg / mL.

[0076] As demonstrated in Examples 9 and 10, the present invention can further improve the extraction rate and purity of gastrodin by modifying the D-101 macroporous adsorption resin. However, the D-101 macroporous adsorption resin itself is a non-polar macroporous adsorption resin and is not suitable for the separation and purification of the polar substance gastrodin.

[0077] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a gastrodin injection, characterized in that, Includes the following steps: (1) 4-Formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside was reduced to 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside; (2) Gastrodin was obtained by deacetylation of 4-hydroxymethylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside; (3) Dissolve gastrodin completely in water for injection, then add pH adjuster and isotonic adjuster, mix evenly, filter, fill, and sterilize to obtain gastrodin injection solution.

2. The method for preparing gastrodin injection according to claim 1, characterized in that: The reduction reaction uses sodium borohydride or potassium borohydride as the reducing agent. Preferably, the amount of the reducing agent is 1 to 2 times the molar amount of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

3. The method for preparing gastrodin injection according to claim 1, characterized in that: The reduction reaction uses barium titanate as a catalyst; Preferably, the amount of catalyst used is 3 to 5% of the amount of 4-formylphenyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside.

4. The method for preparing gastrodin injection according to claim 1, characterized in that: The deacetylation reaction is carried out in a sodium methoxide / methanol or sodium ethoxide / ethanol system.

5. The method for preparing gastrodin injection according to claim 1, characterized in that: The pH adjuster is selected from one or more of the following: acetic acid, hydrochloric acid, phosphoric acid, malic acid, citric acid, tartaric acid, oxalic acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium citrate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate.

6. The method for preparing gastrodin injection according to claim 1, characterized in that: The isotonic regulator is selected from one or more of sodium chloride, glucose, sucrose, lactose, fructose, mannitol, and sorbitol.

7. The method for preparing gastrodin injection according to claim 1, characterized in that: The pH value of the gastrodin injection solution is 5.0~7.

0.

8. The method for preparing gastrodin injection according to claim 1, characterized in that: The osmotic pressure of the gastrodin injection solution is 280~310 mOsm / kg.

9. The method for preparing gastrodin injection according to claim 1, characterized in that: The filtration is membrane filtration; Preferably, the pore size of the filter membrane is 0.22 μm or 0.45 μm.

10. The method for preparing gastrodin injection according to claim 1, characterized in that: The sterilization process is selected from one of the following: autoclaving, dry heat sterilization, and radiation sterilization.

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