Slow-release injection containing levosalbutamol and preparation process thereof

Abstract

The invention provides a slow-release injection containing levosalbutamol and a preparation process thereof, the injection combines a ginkgo leaf extract, a resveratrol extract, vitamin B6, vitamin C, tea polyphenol and other auxiliary components, and through specific preparation steps, the injection can be prepared into the slow-release injection containing levosalbutamol. Comprising the following steps: preparing raw materials, mixing and dissolving, preparing a slow-release carrier, adjusting pH, filtering, filling, sterilizing and the like, thereby obtaining the injection with the slow-release effect. Wherein the ginkgo biloba extract and the resveratrol extract are obtained through specific extraction and purification steps, and the activity and purity of the ginkgo biloba extract and the resveratrol extract are ensured. Polylactic acid-glycolide (PLGA) is used as a sustained-release carrier material and is combined with the active component to form a stable sustained-release system. Every 20 ml of the injection contains 0.2-1 mg of levosalbutamol, and the injection is suitable for a treatment scene where the drug concentration needs to be maintained for a long time. The injection is simple and controllable in preparation process, the obtained product is good in stability and remarkable in slow release effect, and a new choice is provided for treatment of respiratory tract obstructive diseases.

Description

Technical Field

[0001] The invention belongs to the field of biopharmaceuticals and specifically discloses a sustained-release injection containing levosalbutamol and a preparation process thereof. Background Art

[0002] In the medical field, obstructive respiratory diseases such as asthma seriously affect patients' daily life and health. As an effective β2 receptor agonist, levosalbutamol is widely used to treat and prevent such diseases. However, traditional levosalbutamol preparations, such as oral tablets or ordinary injections, have the problem of too fast drug release and short duration of action, which requires patients to take the drug frequently to maintain the treatment effect. This not only increases the patient's medication burden, but also affects the stability and continuity of the treatment effect.

[0003] In the prior art, although there have been some attempts at sustained-release preparations, they often have shortcomings such as complex preparation processes, high costs, unsatisfactory sustained-release effects, or poor drug stability. For example, some sustained-release preparations may use special sustained-release materials or complex preparation processes, which leads to increased production costs, and may introduce unnecessary impurities or pollutants during the production process, affecting the safety and effectiveness of the drug. In addition, the sustained-release effects of some sustained-release preparations are not stable, and the drug release rate is difficult to control, which may lead to fluctuations in the therapeutic effect.

[0004] In addition, the salbutamol preparations in the prior art often lack synergistic effects with other active ingredients, which makes them limited in effect when treating obstructive respiratory diseases. Ingredients such as ginkgo leaf extract and resveratrol extract have multiple biological activities such as anti-oxidation, anti-inflammatory, and bronchial relaxation, and may produce better therapeutic effects when used in combination with salbutamol. However, how to effectively combine these ingredients into the salbutamol preparation and achieve a stable sustained-release effect is a problem to be solved in the prior art.

[0005] In summary, the levosalbutamol preparations in the prior art still have many deficiencies in terms of sustained release effect, drug stability, preparation cost, and synergistic effect with other active ingredients, and need to be further improved and optimized. Summary of the invention

[0006] In order to solve the above problems, we disclose a sustained-release injection containing levosalbutamol and a preparation process thereof.

[0007] The present invention includes the following specific technical solutions:

[0008] A sustained-release injection containing levosalbutamol is prepared from the following raw materials in parts by weight:

[0009]

[0010] Furthermore, in the above-mentioned sustained-release injection containing salbutamol, the ginkgo leaf extract is prepared by the following steps:

[0011] 1) grinding dried ginkgo leaves into powder, and sieving to obtain ginkgo leaf powder of suitable particle size;

[0012] 2) mixing the ginkgo leaf powder with an ethanol solution, wherein the concentration of the ethanol is 40-90%, and subjecting the mixture to microwave ultrasonic treatment for 60-120 minutes at a temperature of 40-50° C., a microwave ultrasonic power of 100-120 W, and a microwave frequency of 40-45 KHz;

[0013] 3) After the extraction is completed, the obtained extract is concentrated until no solvent remains;

[0014] 4) decolorizing the concentrated extract, wherein the decolorizing agent comprises diatomaceous earth, and the mass ratio of the decolorizing agent to the extract is 1-5:100;

[0015] 5) subjecting the decolorized material to acid precipitation treatment, adjusting the pH value to 2-4, and allowing to stand at 2-10° C. for 4-8 hours;

[0016] 6) The liquid after acid precipitation is subjected to column treatment and adsorbed by a macroporous adsorption resin chromatography column. After adsorption, it is first washed with water until the injection liquid is almost colorless, and then eluted with a volume concentration of 60-70% ethanol aqueous solution, the elution rate is 1-3BV / h, and the elution volume is 8-10BV;

[0017] 7) collecting the ethanol eluate, concentrating under reduced pressure and drying to constant weight, and crushing and passing through a 500-mesh sieve to obtain a ginkgo leaf extract.

[0018] Furthermore, in the above-mentioned sustained-release injection containing levosalbutamol, the model of the macroporous adsorption resin is one of ADS-7, AB-8, LK001, polyamide or D-101;

[0019] The mass ratio of the decolorizing agent to the extract is 3:100;

[0020] The concentration of the ethanol is 70%, the microwave ultrasonic treatment time is 90 minutes, the treatment temperature is 45° C., the microwave ultrasonic power is 110 W, and the microwave frequency is 42.5 KHz;

[0021] The acid precipitation treatment is allowed to stand for 6 hours at a pH of 3 and a temperature of 5°C;

[0022] The elution rate is 2BV / h and the elution volume is 9BV.

[0023] Furthermore, in the above-mentioned sustained-release injection containing salbutamol, the resveratrol extract is prepared by the following steps:

[0024] 1) Select fresh or dried plant materials rich in resveratrol, preferably grape skin or Polygonum cuspidatum root, clean them and cut them into pieces of appropriate size;

[0025] 2) placing the chopped plant material in an extraction container, adding an ethanol solution with a concentration of 50%-95%, with a solid-liquid ratio of 1:5 to 1:20 g / ml, performing ultrasonic-assisted extraction at a temperature of 20-60° C., an ultrasonic frequency of 20-60 kHz, and an extraction time of 15-60 minutes;

[0026] 3) After the extraction is completed, filter to remove the solid residue and collect the filtrate;

[0027] 4) Concentrating the filtrate under reduced pressure to remove ethanol and part of the water to obtain a concentrated solution;

[0028] 5) purifying the concentrate by column chromatography, wherein the chromatographic column is filled with silica gel, activated carbon or macroporous resin, and an ethanol solution with a volume fraction of 5% to 30% is used as an eluent, and collecting the eluate containing resveratrol;

[0029] 6) Concentrating the collected eluate under reduced pressure again to remove the solvent to obtain a resveratrol extract;

[0030] 7) Optionally, the extract is subjected to crystallization or recrystallization treatment to improve the purity of resveratrol.

[0031] Furthermore, in the above-mentioned sustained-release injection containing levosalbutamol, the preferred conditions for the ultrasonic-assisted extraction are: ethanol concentration of 70%-80%, extraction temperature of 40-50° C., ultrasonic frequency of 40 kHz, and extraction time of 30-45 minutes;

[0032] In the column chromatography purification step, the chromatography column filler is preferably a macroporous resin, and the volume fraction of ethanol in the eluent is 10%-20%.

[0033] In step 7), a solvent evaporation method or a freeze-drying method is used for crystallization or recrystallization to obtain resveratrol crystals with higher purity.

[0034] Furthermore, in the above-mentioned sustained-release injection containing levosalbutamol, the polylactic acid-glycolide PLGA is copolymerized by lactic acid LA and glycolide GA in a molar ratio of 75:25, and the molecular weight of PLGA is between 5000-30000.

[0035] Furthermore, the above-mentioned sustained-release injection containing levosalbutamol contains 0.2-1 mg of levosalbutamol per 20 ml of the injection.

[0036] The present invention also discloses a preparation process of the sustained-release injection containing levosalbutamol, which comprises the following steps:

[0037] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of levosalbutamol, 3-6 parts of ginkgo leaf extract, 2-4 parts of resveratrol extract, 0.5-1.5 parts of vitamin B6, 1-3 parts of vitamin C, 1-2 parts of tea polyphenols, 5-10 parts of polylactic acid-glycolide PLGA, 4-8 parts of sodium hyaluronate and 100-300 parts of phosphate buffer;

[0038] S2 Mixing and Dissolving:

[0039] Evenly mixing levosalbutamol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols to obtain an active ingredient mixture;

[0040] Adding polylactic acid-co-glycolide PLGA microparticles to the above active ingredient mixture, and uniformly dispersing them by mechanical stirring or ultrasonic treatment;

[0041] Dissolving sodium hyaluronate in an appropriate amount of phosphate buffer to obtain a sodium hyaluronate solution;

[0042] S3 Preparation of sustained-release carrier:

[0043] Slowly add the mixture containing the active ingredient and PLGA microparticles into the sodium hyaluronate solution, stirring while adding to ensure uniform mixing, to form a sustained-release carrier solution;

[0044] S4 pH adjustment and filtration:

[0045] The pH of the sustained-release carrier solution was adjusted to pH 6.8-7.2 using phosphate buffer;

[0046] The sustained-release carrier solution is filtered to remove insoluble impurities and particles therein to ensure the clarity of the injection solution;

[0047] S5 filling and sterilization:

[0048] Filling the filtered sustained-release carrier solution into sterile injection bottles, with each bottle filled with a predetermined amount of injection solution;

[0049] The injection solution after filling is sterilized to achieve the sterility of the injection solution, thereby obtaining the sustained-release injection solution containing levosalbutamol.

[0050] Compared with the prior art, the present invention has the following outstanding beneficial effects:

[0051] 1) Optimized drug combination: By combining multiple active ingredients such as levalbuterol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols, a synergistic effect between drugs is achieved, which is expected to improve the effect of treating obstructive respiratory diseases while reducing the side effects that may be caused by the use of a single drug.

[0052] 2) Stable sustained-release effect: Using polylactic acid-co-glycolide PLGA as a sustained-release material and combining it with sodium hyaluronate to form a sustained-release carrier can effectively control the release rate of the drug, prolong the drug's action time in the body, reduce the patient's medication frequency, and improve the stability and continuity of treatment.

[0053] 3) Improve drug stability: Through specific preparation processes, such as mechanical stirring or ultrasonic treatment to evenly disperse PLGA particles, as well as pH adjustment and filtration treatment, the clarity and stability of the injection are ensured, avoiding drug degradation or aggregation during storage and use.

[0054] 4) Simplified preparation process: The preparation process has clear steps, is easy to operate, and does not require complicated equipment or conditions, which is conducive to reducing production costs, improving production efficiency, and facilitating industrial production.

[0055] 5) Ensure sterility and safety: By filling into sterile injection bottles and sterilizing them, the sterility of the injection solution is ensured, reducing the risk of infection during patient use and improving the safety of the drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 This is the in vitro release test result in the test example. DETAILED DESCRIPTION

[0057] A preparation process of a sustained-release injection containing levosalbutamol comprises the following steps:

[0058] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of levosalbutamol, 3-6 parts of ginkgo leaf extract, 2-4 parts of resveratrol extract, 0.5-1.5 parts of vitamin B6, 1-3 parts of vitamin C, 1-2 parts of tea polyphenols, 5-10 parts of polylactic acid-glycolide PLGA, 4-8 parts of sodium hyaluronate and 100-300 parts of phosphate buffer;

[0059] S2 Mixing and Dissolving:

[0060] Evenly mixing levosalbutamol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols to obtain an active ingredient mixture;

[0061] Adding polylactic acid-co-glycolide PLGA microparticles to the above active ingredient mixture, and uniformly dispersing them by mechanical stirring or ultrasonic treatment;

[0062] Dissolving sodium hyaluronate in an appropriate amount of phosphate buffer to obtain a sodium hyaluronate solution;

[0063] S3 Preparation of sustained-release carrier:

[0064] Slowly add the mixture containing the active ingredient and PLGA microparticles into the sodium hyaluronate solution, stirring while adding to ensure uniform mixing, to form a sustained-release carrier solution;

[0065] S4 pH adjustment and filtration:

[0066] The pH of the sustained-release carrier solution was adjusted to pH 6.8-7.2 using phosphate buffer;

[0067] The sustained-release carrier solution is filtered to remove insoluble impurities and particles therein to ensure the clarity of the injection solution;

[0068] S5 filling and sterilization:

[0069] Filling the filtered sustained-release carrier solution into sterile injection bottles, with each bottle filled with a predetermined amount of injection solution;

[0070] The injection solution after filling is sterilized to achieve the sterility of the injection solution, thereby obtaining the sustained-release injection solution containing levosalbutamol.

[0071] The ginkgo leaf extract is prepared by the following steps:

[0072] 1) grinding dried ginkgo leaves into powder, and sieving to obtain ginkgo leaf powder of suitable particle size;

[0073] 2) mixing the ginkgo leaf powder with an ethanol solution, wherein the concentration of the ethanol is 70%, and subjecting the mixture to microwave ultrasonic treatment for 90 minutes at a temperature of 45° C., a microwave ultrasonic power of 110 W, and a microwave frequency of 42.5 KHz;

[0074] 3) After the extraction is completed, the obtained extract is concentrated until no solvent remains;

[0075] 4) decolorizing the concentrated extract, wherein the decolorizing agent includes diatomaceous earth, and the mass ratio of the decolorizing agent to the extract is 3:100;

[0076] 5) subjecting the decolorized material to acid precipitation, adjusting the pH value to 3, and standing at 5° C. for 6 hours;

[0077] 6) The liquid after acid precipitation is subjected to column treatment and adsorbed with a macroporous adsorption resin chromatography column ADS-7. After adsorption, it is first washed with water until the injection liquid is almost colorless, and then eluted with a volume concentration of 60-70% ethanol aqueous solution, the elution rate is 2BV / h, and the elution volume is 9BV;

[0078] 7) collecting the ethanol eluate, concentrating under reduced pressure and drying to constant weight, and crushing and passing through a 500-mesh sieve to obtain a ginkgo leaf extract.

[0079] The resveratrol extract is prepared by the following steps:

[0080] 1) Selecting dried resveratrol-rich plant material Polygonum cuspidatum root, cleaning it and cutting it into pieces of appropriate size;

[0081] 2) placing the chopped plant material in an extraction container, adding a 70% ethanol solution with a solid-liquid ratio of 1:10 g / ml, performing ultrasonic-assisted extraction at a temperature of 40° C., an ultrasonic frequency of 40 kHz, and an extraction time of 40 minutes;

[0082] 3) After the extraction is completed, filter to remove the solid residue and collect the filtrate;

[0083] 4) Concentrating the filtrate under reduced pressure to remove ethanol and part of the water to obtain a concentrated solution;

[0084] 5) purifying the concentrate by column chromatography, wherein the chromatographic column is filled with a macroporous resin and an ethanol solution with a volume fraction of 15% is used as an eluent, and collecting the eluate containing resveratrol;

[0085] 6) Concentrating the collected eluate under reduced pressure again to remove the solvent to obtain a resveratrol extract;

[0086] 7) The extract is subjected to crystallization or recrystallization treatment to improve the purity of resveratrol.

[0087] The polylactic acid-glycolide PLGA is copolymerized by lactic acid LA and glycolide GA in a molar ratio of 75:25, and the average molecular weight of PLGA is 15,000.

[0088] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0089] Example 1

[0090] A preparation process of a sustained-release injection containing levosalbutamol comprises the following steps:

[0091] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of levosalbutamol, 3 parts of ginkgo leaf extract, 2 parts of resveratrol extract, 0.5 parts of vitamin B6, 1 part of vitamin C, 1 part of tea polyphenols, 5 parts of polylactic acid-co-glycolide PLGA, 4 parts of sodium hyaluronate and 100 parts of phosphate buffer;

[0092] S2 Mixing and Dissolving:

[0093] Evenly mixing levosalbutamol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols to obtain an active ingredient mixture;

[0094] Adding polylactic acid-co-glycolide PLGA microparticles to the above active ingredient mixture, and uniformly dispersing them by mechanical stirring or ultrasonic treatment;

[0095] Dissolving sodium hyaluronate in an appropriate amount of phosphate buffer to obtain a sodium hyaluronate solution;

[0096] S3 Preparation of sustained-release carrier:

[0097] Slowly add the mixture containing the active ingredient and PLGA microparticles into the sodium hyaluronate solution, stirring while adding to ensure uniform mixing, to form a sustained-release carrier solution;

[0098] S4 pH adjustment and filtration:

[0099] The pH of the sustained-release carrier solution was adjusted to pH 6.8-7.2 using phosphate buffer;

[0100] The sustained-release carrier solution is filtered to remove insoluble impurities and particles therein to ensure the clarity of the injection solution;

[0101] S5 filling and sterilization:

[0102] Filling the filtered sustained-release carrier solution into sterile injection bottles, with each bottle filled with a predetermined amount of injection solution;

[0103] The injection solution after filling is sterilized to achieve the sterility of the injection solution, thereby obtaining the sustained-release injection solution containing levosalbutamol.

[0104] The ginkgo leaf extract is prepared by the following steps:

[0105] 1) grinding dried ginkgo leaves into powder, and sieving to obtain ginkgo leaf powder of suitable particle size;

[0106] 2) mixing the ginkgo leaf powder with an ethanol solution, wherein the concentration of the ethanol is 70%, and subjecting the mixture to microwave ultrasonic treatment for 90 minutes at a temperature of 45° C., a microwave ultrasonic power of 110 W, and a microwave frequency of 42.5 KHz;

[0107] 3) After the extraction is completed, the obtained extract is concentrated until no solvent remains;

[0108] 4) decolorizing the concentrated extract, wherein the decolorizing agent includes diatomaceous earth, and the mass ratio of the decolorizing agent to the extract is 3:100;

[0109] 5) subjecting the decolorized material to acid precipitation, adjusting the pH value to 3, and standing at 5° C. for 6 hours;

[0110] 6) The liquid after acid precipitation is subjected to column treatment and adsorbed with a macroporous adsorption resin chromatography column ADS-7. After adsorption, it is first washed with water until the injection liquid is almost colorless, and then eluted with a volume concentration of 60-70% ethanol aqueous solution, the elution rate is 2BV / h, and the elution volume is 9BV;

[0111] 7) collecting the ethanol eluate, concentrating under reduced pressure and drying to constant weight, and crushing and passing through a 500-mesh sieve to obtain a ginkgo leaf extract.

[0112] The resveratrol extract is prepared by the following steps:

[0113] 1) Selecting dried resveratrol-rich plant material Polygonum cuspidatum root, cleaning it and cutting it into pieces of appropriate size;

[0114] 2) placing the chopped plant material in an extraction container, adding a 70% ethanol solution with a solid-liquid ratio of 1:10 g / ml, performing ultrasonic-assisted extraction at a temperature of 40° C., an ultrasonic frequency of 40 kHz, and an extraction time of 40 minutes;

[0115] 3) After the extraction is completed, filter to remove the solid residue and collect the filtrate;

[0116] 4) Concentrating the filtrate under reduced pressure to remove ethanol and part of the water to obtain a concentrated solution;

[0117] 5) purifying the concentrate by column chromatography, wherein the chromatographic column is filled with a macroporous resin and an ethanol solution with a volume fraction of 15% is used as an eluent, and collecting the eluate containing resveratrol;

[0118] 6) Concentrating the collected eluate under reduced pressure again to remove the solvent to obtain a resveratrol extract;

[0119] 7) The extract is subjected to crystallization or recrystallization treatment to improve the purity of resveratrol.

[0120] The polylactic acid-glycolide PLGA is copolymerized by lactic acid LA and glycolide GA in a molar ratio of 75:25, and the average molecular weight of PLGA is 15,000.

[0121] Example 2

[0122] A preparation process of a sustained-release injection containing levosalbutamol comprises the following steps:

[0123] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of salbutamol, 4.5 parts of ginkgo leaf extract, 3 parts of resveratrol extract, 1.0 parts of vitamin B6, 2.0 parts of vitamin C, 1.5 parts of tea polyphenols, 7.5 parts of polylactic acid-glycolide PLGA, 6 parts of sodium hyaluronate and 200 parts of phosphate buffer;

[0124] S2 Mixing and Dissolving:

[0125] Evenly mixing levosalbutamol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols to obtain an active ingredient mixture;

[0126] Adding polylactic acid-co-glycolide PLGA microparticles to the above active ingredient mixture, and uniformly dispersing them by mechanical stirring or ultrasonic treatment;

[0127] Dissolving sodium hyaluronate in an appropriate amount of phosphate buffer to obtain a sodium hyaluronate solution;

[0128] S3 Preparation of sustained-release carrier:

[0129] Slowly add the mixture containing the active ingredient and PLGA microparticles into the sodium hyaluronate solution, stirring while adding to ensure uniform mixing, to form a sustained-release carrier solution;

[0130] S4 pH adjustment and filtration:

[0131] The pH of the sustained-release carrier solution was adjusted to pH 6.8-7.2 using phosphate buffer;

[0132] The sustained-release carrier solution is filtered to remove insoluble impurities and particles therein to ensure the clarity of the injection solution;

[0133] S5 filling and sterilization:

[0134] Filling the filtered sustained-release carrier solution into sterile injection bottles, with each bottle filled with a predetermined amount of injection solution;

[0135] The injection solution after filling is sterilized to achieve the sterility of the injection solution, thereby obtaining the sustained-release injection solution containing levosalbutamol.

[0136] The ginkgo leaf extract is prepared by the following steps:

[0137] 1) grinding dried ginkgo leaves into powder, and sieving to obtain ginkgo leaf powder of suitable particle size;

[0138] 2) mixing the ginkgo leaf powder with an ethanol solution, wherein the concentration of the ethanol is 70%, and subjecting the mixture to microwave ultrasonic treatment for 90 minutes at a temperature of 45° C., a microwave ultrasonic power of 110 W, and a microwave frequency of 42.5 KHz;

[0139] 3) After the extraction is completed, the obtained extract is concentrated until no solvent remains;

[0140] 4) decolorizing the concentrated extract, wherein the decolorizing agent includes diatomaceous earth, and the mass ratio of the decolorizing agent to the extract is 3:100;

[0141] 5) subjecting the decolorized material to acid precipitation, adjusting the pH value to 3, and standing at 5° C. for 6 hours;

[0142] 6) The liquid after acid precipitation is subjected to column treatment and adsorbed with a macroporous adsorption resin chromatography column ADS-7. After adsorption, it is first washed with water until the injection liquid is almost colorless, and then eluted with a volume concentration of 60-70% ethanol aqueous solution, the elution rate is 2BV / h, and the elution volume is 9BV;

[0143] 7) collecting the ethanol eluate, concentrating under reduced pressure and drying to constant weight, and crushing and passing through a 500-mesh sieve to obtain a ginkgo leaf extract.

[0144] The resveratrol extract is prepared by the following steps:

[0145] 1) Selecting dried resveratrol-rich plant material Polygonum cuspidatum root, cleaning it and cutting it into pieces of appropriate size;

[0146] 2) placing the chopped plant material in an extraction container, adding a 70% ethanol solution with a solid-liquid ratio of 1:10 g / ml, performing ultrasonic-assisted extraction at a temperature of 40° C., an ultrasonic frequency of 40 kHz, and an extraction time of 40 minutes;

[0147] 3) After the extraction is completed, filter to remove the solid residue and collect the filtrate;

[0148] 4) Concentrating the filtrate under reduced pressure to remove ethanol and part of the water to obtain a concentrated solution;

[0149] 5) purifying the concentrate by column chromatography, wherein the chromatographic column is filled with a macroporous resin and an ethanol solution with a volume fraction of 15% is used as an eluent, and collecting the eluate containing resveratrol;

[0150] 6) Concentrating the collected eluate under reduced pressure again to remove the solvent to obtain a resveratrol extract;

[0151] 7) The extract is subjected to crystallization or recrystallization treatment to improve the purity of resveratrol.

[0152] The polylactic acid-glycolide PLGA is copolymerized by lactic acid LA and glycolide GA in a molar ratio of 75:25, and the average molecular weight of PLGA is 15,000.

[0153] Example 3

[0154] A preparation process of a sustained-release injection containing levosalbutamol comprises the following steps:

[0155] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of salbutamol, 6 parts of ginkgo leaf extract, 4 parts of resveratrol extract, 1.5 parts of vitamin B6, 3 parts of vitamin C, 2 parts of tea polyphenols, 10 parts of polylactic acid-glycolide PLGA, 8 parts of sodium hyaluronate and 300 parts of phosphate buffer;

[0156] S2 Mixing and Dissolving:

[0157] Evenly mixing levosalbutamol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols to obtain an active ingredient mixture;

[0158] Adding polylactic acid-co-glycolide PLGA microparticles to the above active ingredient mixture, and uniformly dispersing them by mechanical stirring or ultrasonic treatment;

[0159] Dissolving sodium hyaluronate in an appropriate amount of phosphate buffer to obtain a sodium hyaluronate solution;

[0160] S3 Preparation of sustained-release carrier:

[0161] Slowly add the mixture containing the active ingredient and PLGA microparticles into the sodium hyaluronate solution, stirring while adding to ensure uniform mixing, to form a sustained-release carrier solution;

[0162] S4 pH adjustment and filtration:

[0163] The pH of the sustained-release carrier solution was adjusted to pH 6.8-7.2 using phosphate buffer;

[0164] The sustained-release carrier solution is filtered to remove insoluble impurities and particles therein to ensure the clarity of the injection solution;

[0165] S5 filling and sterilization:

[0166] Filling the filtered sustained-release carrier solution into sterile injection bottles, with each bottle filled with a predetermined amount of injection solution;

[0167] The injection solution after filling is sterilized to achieve the sterility of the injection solution, thereby obtaining the sustained-release injection solution containing levosalbutamol.

[0168] The ginkgo leaf extract is prepared by the following steps:

[0169] 1) grinding dried ginkgo leaves into powder, and sieving to obtain ginkgo leaf powder of suitable particle size;

[0170] 2) mixing the ginkgo leaf powder with an ethanol solution, wherein the concentration of the ethanol is 70%, and subjecting the mixture to microwave ultrasonic treatment for 90 minutes at a temperature of 45° C., a microwave ultrasonic power of 110 W, and a microwave frequency of 42.5 KHz;

[0171] 3) After the extraction is completed, the obtained extract is concentrated until no solvent remains;

[0172] 4) decolorizing the concentrated extract, wherein the decolorizing agent includes diatomaceous earth, and the mass ratio of the decolorizing agent to the extract is 3:100;

[0173] 5) subjecting the decolorized material to acid precipitation, adjusting the pH value to 3, and standing at 5° C. for 6 hours;

[0174] 6) The liquid after acid precipitation is subjected to column treatment and adsorbed with a macroporous adsorption resin chromatography column ADS-7. After adsorption, it is first washed with water until the injection liquid is almost colorless, and then eluted with a volume concentration of 60-70% ethanol aqueous solution, the elution rate is 2BV / h, and the elution volume is 9BV;

[0175] 7) collecting the ethanol eluate, concentrating under reduced pressure and drying to constant weight, and crushing and passing through a 500-mesh sieve to obtain a ginkgo leaf extract.

[0176] The resveratrol extract is prepared by the following steps:

[0177] 1) Selecting dried resveratrol-rich plant material Polygonum cuspidatum root, cleaning it and cutting it into pieces of appropriate size;

[0178] 2) placing the chopped plant material in an extraction container, adding a 70% ethanol solution with a solid-liquid ratio of 1:10 g / ml, performing ultrasonic-assisted extraction at a temperature of 40° C., an ultrasonic frequency of 40 kHz, and an extraction time of 40 minutes;

[0179] 3) After the extraction is completed, filter to remove the solid residue and collect the filtrate;

[0180] 4) Concentrating the filtrate under reduced pressure to remove ethanol and part of the water to obtain a concentrated solution;

[0181] 5) purifying the concentrate by column chromatography, wherein the chromatographic column is filled with a macroporous resin and an ethanol solution with a volume fraction of 15% is used as an eluent, and collecting the eluate containing resveratrol;

[0182] 6) Concentrating the collected eluate under reduced pressure again to remove the solvent to obtain a resveratrol extract;

[0183] 7) The extract is subjected to crystallization or recrystallization treatment to improve the purity of resveratrol.

[0184] The polylactic acid-glycolide PLGA is copolymerized by lactic acid LA and glycolide GA in a molar ratio of 75:25, and the average molecular weight of PLGA is 15,000.

[0185] Comparative Example 1

[0186] A preparation process of a sustained-release injection containing levosalbutamol comprises the following steps:

[0187] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of salbutamol, 4 parts of resveratrol extract, 1.5 parts of vitamin B6, 3 parts of vitamin C, 2 parts of tea polyphenols, 10 parts of polylactic acid-co-glycolide PLGA, 8 parts of sodium hyaluronate and 300 parts of phosphate buffer;

[0188] The other parts are the same as those in Example 2 except that the ginkgo leaf extract is not contained.

[0189] Comparative Example 2

[0190] A preparation process of a sustained-release injection containing levosalbutamol comprises the following steps:

[0191] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of levosalbutamol, 6 parts of ginkgo leaf extract, 1.5 parts of vitamin B6, 3 parts of vitamin C, 2 parts of tea polyphenols, 10 parts of polylactic acid-glycolide PLGA, 8 parts of sodium hyaluronate and 300 parts of phosphate buffer;

[0192] The resveratrol extract is not contained, and the rest is the same as in Example 2.

[0193] Comparative Example 3

[0194] A preparation process of a sustained-release injection containing levosalbutamol comprises the following steps:

[0195] S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of levosalbutamol, 6 parts of ginkgo leaf extract, 4 parts of resveratrol extract, 1.5 parts of vitamin B6, 3 parts of vitamin C, 2 parts of tea polyphenols, and 300 parts of phosphate buffer;

[0196] It does not contain polylactic acid-co-glycolide PLGA and sodium hyaluronate, and the rest is the same as Example 2.

[0197] Test Example 1

[0198] The sustained-release injections containing levosalbutamol prepared in the above Examples 1-3 and Comparative Examples 1-3, and the commercially available salbutamol sulfate injection (Comparative Example 4) were tested together for comparative testing, and the testing was conducted with reference to the pharmacopoeia standards.

[0199] Table 1 Injection properties test

[0200]

[0201]

[0202] It can be seen from the data in Table 1 above that after adding polylactic acid-co-glycolide PLGA and sodium hyaluronate, the shape test of the present invention can meet the requirements, reduce the floccules in the solution, and reduce the detection of impurities.

[0203] Test Example 2

[0204] The stability test was carried out on the sustained-release injection containing levosalbutamol prepared in the above Examples 1-3 and Comparative Examples 1-3, and the commercially available salbutamol sulfate injection (Comparative Example 4). The stability test was carried out at 60°C and light (1.2×10 6 Lux·hr, near ultraviolet not less than 200W·hr / m 2 The accelerated stability was investigated for 1 and 6 months under the conditions of . The results are shown in Table 2-4.

[0205] Table 2. 1-month accelerated stability

[0206]

[0207] Table 3. 6-month accelerated stability

[0208]

[0209] It can be seen from Tables 2 and 3 above that the addition of ginkgo leaf extract and resveratrol extract can greatly increase the stability of the sustained-release injection.

[0210] Test Example 3

[0211] In vitro release test:

[0212] The in vitro release behavior of the sustained-release injection containing levosalbutamol prepared in Examples 1-3 and Comparative Examples 1-3, and the commercially available salbutamol sulfate injection (Comparative Example 4) was investigated by the constant temperature shaking table method: refer to the method in CN201910804352.5. The results are shown in Tables 4 and Figure 1shown.

[0213] Table 4 In vitro release test

[0214]

[0215] From the results in Table 4, it can be seen that the addition of polylactic acid-co-glycolide PLGA and sodium hyaluronate can achieve a smooth in vitro release with a significant sustained release effect. From Comparative Examples 1 and 2, it can also be seen that the addition of ginkgo leaf extract and resveratrol extract can help improve the sustained release effect.

[0216] As can be seen from the above examples, the present invention achieves synergy between drugs by combining multiple active ingredients such as salbutamol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols, and is expected to improve the effect of treating obstructive respiratory diseases, while reducing the side effects that may be caused by the use of a single drug. Using polylactic acid-co-glycolide PLGA as a sustained-release material and combining sodium hyaluronate to form a sustained-release carrier can effectively control the release rate of the drug, prolong the action time of the drug in the body, reduce the frequency of medication for patients, and improve the stability and continuity of treatment. Through a specific preparation process, such as mechanical stirring or ultrasonic treatment to uniformly disperse PLGA particles, and adjusting the pH value and filtering treatment, the clarity and stability of the injection are ensured, and the degradation or aggregation of the drug during storage and use is avoided.

[0217] The above are only a few preferred embodiments of the present invention, which are described in a more specific and detailed manner, but they cannot be understood as limiting the scope of the present invention. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A sustained-release injection containing salbutamol, characterized in that: Made from the following raw materials by weight: 10 parts of Levosalbutamol 3-6 servings of Ginkgo biloba extract 2-4 servings of Resveratrol Extract Vitamin B6 0.5-1.5 parts Vitamin C 1-3 servings 1-2 servings of tea polyphenols Polylactic acid-glycolide PLGA 5-10 parts 4-8 parts of sodium hyaluronate Phosphate buffer 100-300 parts.

2. A sustained-release injection containing levosalbutamol according to claim 1, characterized in that: The ginkgo leaf extract is prepared by the following steps: 1) Grinding the dried ginkgo leaves into powder, and sieving to obtain ginkgo leaf powder of suitable particle size; 2) mixing the ginkgo leaf powder with an ethanol solution, wherein the concentration of the ethanol is 40-90%, and subjecting the mixture to microwave ultrasonic treatment for 60-120 minutes at a temperature of 40-50° C., a microwave ultrasonic power of 100-120 W, and a microwave frequency of 40-45 KHz; 3) After the extraction is completed, the obtained extract is concentrated until no solvent remains; 4) Decolorizing the concentrated extract, wherein the decolorizing agent includes diatomaceous earth, and the mass ratio of the decolorizing agent to the extract is 1-5:100; 5) Treat the decolorized material with acid precipitation, adjust the pH value to 2-4, and let it stand at 2-10°C for 4-8 hours; 6) The liquid after acid precipitation is subjected to column treatment and adsorbed with a macroporous adsorption resin chromatography column. After adsorption, it is first washed with water until the injection liquid is almost colorless, and then eluted with a volume concentration of 60-70% ethanol aqueous solution. The elution rate is 1-3BV / h and the elution volume is 8-10BV; 7) Collect the ethanol eluate, concentrate under reduced pressure and dry to constant weight, and grind it through a 500-mesh sieve to obtain the ginkgo leaf extract.

3. The sustained-release injection containing salbutamol according to claim 2, characterized in that: The model of the macroporous adsorption resin is one of ADS-7, AB-8, LK001, polyamide or D-101; The mass ratio of the decolorizing agent to the extract is 3:100; The concentration of ethanol is 70%, the microwave ultrasonic treatment time is 90 minutes, the treatment temperature is 45° C., the microwave ultrasonic power is 110 W, and the microwave frequency is 42.5 KHz; The acid precipitation treatment is allowed to stand for 6 hours at a pH of 3 and a temperature of 5°C; The elution rate is 2BV / h and the elution volume is 9BV.

4. The sustained-release injection containing levosalbutamol according to claim 1, characterized in that: The resveratrol extract is prepared by the following steps: 1) Select fresh or dried plant materials rich in resveratrol, preferably grape skin or knotweed root, clean them and cut them into pieces of appropriate size; 2) Place the chopped plant material in an extraction container, add 50%-95% ethanol solution, with a solid-liquid ratio of 1:5 to 1:20 g / ml, perform ultrasonic-assisted extraction at a temperature of 20-60°C, an ultrasonic frequency of 20-60kHz, and an extraction time of 15-60 minutes; 3) After the extraction is completed, filter to remove the solid residue and collect the filtrate; 4) Concentrating the filtrate under reduced pressure to remove ethanol and part of the water to obtain a concentrated solution; 5) purifying the concentrate by column chromatography, wherein the chromatography column is filled with silica gel, activated carbon or macroporous resin, and an ethanol solution with a volume fraction of 5%-30% is used as an eluent, and collecting the eluate containing resveratrol; 6) Concentrating the collected eluate under reduced pressure again to remove the solvent to obtain a resveratrol extract; 7) Optionally, the extract is subjected to crystallization or recrystallization treatment to improve the purity of resveratrol.

5. The sustained-release injection containing levosalbutamol according to claim 1, characterized in that: The preferred conditions for the ultrasonic-assisted extraction are: ethanol concentration of 70%-80%, extraction temperature of 40-50 ℃, ultrasonic frequency of 40kHz, extraction time of 30-45 minutes; In the column chromatography purification step, the chromatography column filler is preferably a macroporous resin, and the volume fraction of ethanol in the eluent is 10%-20%; In step 7), a solvent evaporation method or a freeze-drying method is used for crystallization or recrystallization to obtain resveratrol crystals with higher purity.

6. The sustained-release injection containing levosalbutamol according to claim 1, characterized in that: The polylactic acid-glycolide PLGA is copolymerized by lactic acid LA and glycolide GA in a molar ratio of 75:25, and the molecular weight of PLGA is between 5000-30000.

7. The sustained-release injection containing levosalbutamol according to claim 1, characterized in that: Each 20 ml of the injection contains 0.2-1 mg of levosalbutamol.

8. The process for preparing the sustained-release injection containing levosalbutamol according to any one of claims 1 to 7, characterized in that: The following steps are involved: S1. Raw material preparation: prepare the following raw materials by weight: 10 parts of levosalbutamol, 3-6 parts of ginkgo leaf extract, 2-4 parts of resveratrol extract, 0.5-1.5 parts of vitamin B6, 1-3 parts of vitamin C, 1-2 parts of tea polyphenols, 5-10 parts of polylactic acid-glycolide PLGA, 4-8 parts of sodium hyaluronate and 100-300 parts of phosphate buffer; S2 Mixing and Dissolving: Evenly mixing levosalbutamol, ginkgo leaf extract, resveratrol extract, vitamin B6, vitamin C and tea polyphenols to obtain an active ingredient mixture; Adding polylactic acid-co-glycolide PLGA microparticles to the above active ingredient mixture, and uniformly dispersing them by mechanical stirring or ultrasonic treatment; Dissolving sodium hyaluronate in an appropriate amount of phosphate buffer to obtain a sodium hyaluronate solution; S3 Preparation of sustained-release carrier: Slowly add the mixture containing the active ingredient and PLGA microparticles into the sodium hyaluronate solution, stirring while adding to ensure uniform mixing, to form a sustained-release carrier solution; S4 pH adjustment and filtration: The pH of the sustained-release carrier solution was adjusted to pH 6.8-7.2 using phosphate buffer; The sustained-release carrier solution is filtered to remove insoluble impurities and particles therein to ensure the clarity of the injection solution; S5 filling and sterilization: Filling the filtered sustained-release carrier solution into sterile injection bottles, with each bottle filled with a predetermined amount of injection solution; The injection solution after filling is sterilized to achieve the sterility of the injection solution, thereby obtaining the sustained-release injection solution containing levosalbutamol.

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