Traditional Chinese medicine injection containing dendritic macromolecular polysaccharide and preparation method thereof

Abstract

The invention discloses a Chinese medicine injection containing dendritic macromolecular polysaccharide and a preparation method thereof, and belongs to the technical field of Chinese medicine preparations. Dendritic macromolecular polysaccharide, two molecular weight polysaccharides of 1KD~10KD and 10KD~100KD are mixed at a mass ratio of 1:3. The Chinese medicine injection in which the dendritic macromolecular polysaccharide replaces Tween 80 comprises 200mL of a concentrated solution of the dendritic macromolecular polysaccharide, 200mL of an injection alcohol extract concentrate, 0.1~0.5wt% of an antioxidant, a pH regulator and water for injection, and the pH is adjusted to 7~8 to prepare a 1000mL solution. The dendritic macromolecular polysaccharide of the invention is a polysaccharide in the preparation prescription itself, which can effectively maintain the stability of the Chinese medicine injection system.

Description

Technical Field

[0001] The present invention relates to a Chinese medicine injection containing a dendritic macromolecular polysaccharide and a preparation method thereof, and in particular to a novel solubilizing agent polysaccharide, which is used to replace Tween 80 to maintain the stability of the Chinese medicine injection and has better hemolytic safety and allergic-like safety, and belongs to the technical field of Chinese medicine preparations. Background Art

[0002] Chinese medicine injection is a liquid preparation obtained by extracting, separating and purifying Chinese medicine slices and specially injected into the body. Chinese medicine injection breaks through the traditional way of administering Chinese medicine and has the characteristics of high bioavailability and rapid efficacy. However, the composition of Chinese medicine injection is relatively complex, and there are many components with low solubility, such as berberine and ginseng secondary saponin, which are common insoluble effective components. In the preparation process, in order to dissolve insoluble components, it is generally chosen to increase the amount of solvent or use solubilization technology. However, the use of a large amount of solvent not only violates the daily use habits of injection, but also causes systemic edema, venous inflammation, organ damage, and may even be life-threatening. Therefore, choosing appropriate solubilization technology according to the characteristics of the effective components is the first choice to maintain the stability and safety of Chinese medicine injection.

[0003] In industrial production, adding solubilizer Tween 80 is an important method to maintain the clarity and stability of the injection system. Tween 80 is a non-ionic surfactant that can form micelles in water, encapsulate and adsorb the insoluble components in the traditional Chinese medicine injection, improve the solubility, and make the injection system clear and stable. However, Tween 80 may cause a series of adverse reactions such as hemolysis, allergy, allergic reactions, and peripheral neurotoxicity. Animal experiments have shown that after intravenous infusion of Tween 80 solution and traditional Chinese medicine injection containing Tween 80 to Beagle dogs, Beagle dogs have allergic reactions such as decreased blood pressure, slowed heart rate, and skin erythema. The allergic mechanism may be to accelerate vesicle circulation by increasing cell membrane fluidity, promote cell degranulation, and increase the release of histamine and β-hexosaminidase to cause allergic reaction symptoms.

[0004] The prescription of Shenmai injection comes from "Shengmai Powder" in "Prescriptions for Emergencies" written by Sun Simiao. It is mainly composed of two Chinese medicines, Ophiopogon japonicus and red ginseng. It has the effects of invigorating qi and consolidating deficiency, nourishing yin and promoting body fluid, and promoting pulse. It is clinically used to treat shock caused by qi and yin deficiency, chronic heart failure, coronary heart disease, viral myocarditis, chronic cor pulmonale, granulocytopenia, and can improve the immunity of tumor patients. However, the ginseng secondary saponins produced by high-temperature sterilization during the production process have low solubility and are easily precipitated from the solution, affecting the stability of the injection system. Therefore, there is an urgent need for a safe and non-toxic dendritic macromolecular polysaccharide that can maintain the stability of the injection system and has better hemolytic safety and allergic safety, and a Chinese medicine injection, preparation method and application thereof that replaces Tween 80. Summary of the invention

[0005] The purpose of the present invention is to address the problems existing in the prior art and provide a dendritic macromolecular polysaccharide as a new solubilizing agent for replacing Tween 80 to maintain the stability of traditional Chinese medicine injections, and having better hemolytic safety and allergic-like safety.

[0006] Another object of the present invention is to provide a method for preparing dendritic macromolecular polysaccharide.

[0007] Another object of the present invention is to provide a traditional Chinese medicine injection in which dendritic macromolecular polysaccharide replaces Tween 80.

[0008] Another object of the present invention is to provide a method for preparing a traditional Chinese medicine injection in which dendritic macromolecular polysaccharide replaces Tween 80.

[0009] Another object of the present invention is to provide an application of dendritic macromolecular polysaccharide.

[0010] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0011] Dendritic macromolecular polysaccharide, two molecular weight polysaccharides of 1KD-10KD and 10KD-100KD are mixed in a mass ratio of 1:3.

[0012] The preparation method of dendritic macromolecular polysaccharide comprises the following steps: using 3-10 BV 50-90% ethanol at 70-90° C. to extract Chinese herbal medicine pieces 3-9 times, each time for 1-5 hours, drying the obtained Chinese herbal medicine alcohol extraction residues at normal pressure in a drying oven at 50-60° C., adding 5-10 BV purified water or 10% sodium hydroxide solution cold water to extract the dried Chinese herbal medicine alcohol extraction residues 2-3 times, each extraction time is 30-120 minutes, obtaining a Chinese herbal medicine crude polysaccharide water extract, centrifuging at 1000-12000 rpm for 5-30 minutes, taking the supernatant and using hollow fiber column ultrafiltration to prepare Chinese herbal medicine polysaccharide, collecting component A: 1KD-10KD and component B: 10KD-100KD Chinese herbal medicine polysaccharide solutions, respectively, reducing pressure and concentrating component A and component B to a relative density of 1.1 0~1.30, vacuum concentration degree -0.1~0MPa, concentration temperature 60~90℃, 95% pure industrial ethanol is added to concentrated Chinese medicine extracts A and B respectively, stirring while adding until the alcohol concentration ranges from 50% to 90%, and refrigerated and stored at 0~4℃ for 6~12h, and the precipitate and supernatant of the alcohol precipitate are separated by suction filtration, and the precipitate is washed with 95% industrial ethanol until the supernatant is colorless, and the precipitate is dried under reduced pressure to obtain Chinese medicine polysaccharides A and B, the molecular weight of Chinese medicine polysaccharide A is 1KD~10KD, and the molecular weight of Chinese medicine polysaccharide B is 10KD~100KD, and Chinese medicine polysaccharide A and Chinese medicine polysaccharide B are mixed in a mass ratio of 1:3 to obtain dendritic macromolecular polysaccharide.

[0013] Chinese herbal medicines include red ginseng slices, licorice slices and epimedium slices.

[0014] The method for preparing a concentrated solution of dendritic macromolecular polysaccharide comprises the following steps: taking a dendritic macromolecular polysaccharide, preparing a concentrated solution of the dendritic macromolecular polysaccharide with a concentration of 1 to 10 g / L, adjusting the pH value of the solution to 1 to 4 using a hydrochloric acid solution, heating and boiling the solution for 10 to 30 minutes, cooling the solution and centrifuging the solution at 1000 to 12000 rpm for 5 to 30 minutes to obtain a supernatant, adjusting the pH value of the supernatant solution to 9 to 14 using a sodium hydroxide solution, heating and boiling the solution for 10 to 30 minutes, cooling the solution and centrifuging the solution at 1000 to 12000 rpm for 5 to 30 minutes to obtain a supernatant, and filtering the solution using a 0.45 μm microporous filter membrane to obtain a pretreated concentrated solution of the dendritic macromolecular polysaccharide.

[0015] The traditional Chinese medicine injection in which dendritic macromolecular polysaccharide replaces Tween 80 comprises 200 mL of a concentrated solution of the dendritic macromolecular polysaccharide, 200 mL of an injection alcohol extract concentrate, 0.1-0.5 wt % of an antioxidant, a pH regulator and water for injection, and the pH is adjusted to 7-8 to prepare a 1000 mL solution.

[0016] The antioxidant includes vitamin C, sodium bisulfite or sodium pyrosulfite; the pH regulator is 10% sodium hydroxide; and the injection water is the injection water that is boiled and deoxygenated.

[0017] The preparation method of the alcohol extract concentrate for injection is as follows: taking the amount of red ginseng and ophiopogon japonicus used in shengmai injection, respectively adding 3-10BV 50-90% ethanol at 70-90°C to extract 3-9 times, each time for 1-5 hours, collecting the extract, reducing pressure to recover ethanol until each 1mL contains 0.3-0.4g of crude drug, adding 1-10% activated carbon and stirring for 0.5-3h, reducing pressure to concentrate until there is no alcohol taste, adding water for injection until each 1mL contains 2.0g of crude drug, filtering and mixing to obtain the alcohol extract concentrate for shengmai injection.

[0018] The preparation method of a traditional Chinese medicine injection in which dendritic macromolecular polysaccharide replaces Tween 80 comprises the following steps: taking 200 mL of injection alcohol extraction concentrate, adding 200 mL of a dendritic macromolecular polysaccharide concentrated solution, adding 0.1-0.5 wt% of an antioxidant, adding boiling and deoxygenated injection water to prepare a 1000 mL solution, adjusting the pH to 6-8 with a pH regulator, filling in an ampoule under N2 protection, and dry-heat sterilizing at 115-121°C for 20-31 minutes to obtain a shenmai injection in which the dendritic macromolecular polysaccharide replaces Tween 80.

[0019] Application of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0020] Application of concentrated solution of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0021] As one of the main ingredients of traditional Chinese medicine, polysaccharides have a variety of chain conformations, good water solubility, large molecular weight, high degree of branching, micelle formation, easy cross-linking and other characteristics. They can reduce the intramolecular effects of poorly soluble molecules or reduce the surface tension of water molecules, and play a solubilizing effect different from the cyclodextrin mechanism. Moreover, as a natural ingredient, polysaccharides have the characteristics of low toxicity and biocompatibility, and have great potential in replacing the traditional solubilizer Tween 80 for traditional Chinese medicine injections.

[0022] The polysaccharide of the present invention can increase the solubility of poorly soluble drugs, so the red ginseng polysaccharide from the Shenmai prescription itself is an ideal material for the combination of medicine and adjuvant in Chinese medicine injection. Therefore, the present invention uses the polysaccharide in the Chinese medicine injection prescription itself to replace Tween 80 as a solubilizer, and prepares a Chinese medicine injection in which polysaccharide replaces Tween 80, which can maintain the stability of the injection system and has better hemolytic safety and allergic safety.

[0023] In summary, the present invention provides a Chinese medicine injection in which a dendritic macromolecular polysaccharide replaces a solubilizing agent Tween 80 and a preparation method thereof, which can maintain the stability of the injection system. The method of the present invention has the following advantages: based on the holistic concept of the substance basis of traditional Chinese medicine, the original prescription Chinese medicine polysaccharide is used as a solubilizing agent to replace Tween 80 to maintain the stability of the injection, and the Chinese medicine polysaccharide has low toxicity, high biocompatibility, and higher safety; the alcohol-extracted drug residue can be recycled and reused, saving a lot of manpower and material resources, creating huge economic and social benefits, and is of great significance to promoting the development of the integration of Chinese medicine preparation products and adjuvants.

[0024] The present invention provides a Chinese medicine injection in which a dendritic macromolecular polysaccharide replaces Tween 80 and a preparation method thereof. The Chinese medicine injection in which a dendritic macromolecular polysaccharide replaces Tween 80 of the present invention comprises a dendritic macromolecular polysaccharide, an injection alcohol extract concentrate, an antioxidant, a pH regulator and water for injection; the dendritic macromolecular polysaccharide is a polysaccharide in the preparation prescription itself; the preparation method comprises: extracting the dendritic macromolecular polysaccharide, preparing the injection solution, an antioxidant and a solubilizing agent are uniformly mixed, the pH is adjusted, and the injection water from which oxygen is removed is used to make a fixed volume and nitrogen is added for filling and sterilization. The dendritic macromolecular polysaccharide replacing Tween 80 of the present invention can effectively maintain the stability of the Chinese medicine injection system. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 TEM image of red ginseng dendrimer polysaccharide;

[0026] Figure 2 TEM image of red ginseng polysaccharide extracted from water and precipitated with alcohol. DETAILED DESCRIPTION

[0027] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments. The following embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0028] Example 1

[0029] The red ginseng pieces were extracted five times with 3BV 50% ethanol at 90℃ for 1h, and the alcohol extract was used to prepare Shenmai injection. The red ginseng alcohol-extracted residues were dried at normal pressure in a drying oven at 50℃. The dried red ginseng alcohol-extracted residues were then extracted twice with 5BV purified water, each time for 60min, to obtain a crude red ginseng polysaccharide water extract, which was centrifuged at 3000rpm for 10min, and the supernatant was ultrafiltered using a hollow fiber column to prepare red ginseng polysaccharides, and component A: 1KD~10KD and component B: 10KD~100KD red ginseng polysaccharide solution was collected. liquid, respectively, reducing pressure and concentrating component A and component B to a relative density of 1.10, the vacuum degree of reduced pressure and concentration is -0.1MPa, the concentration temperature is 90°C, 95% pure industrial ethanol is slowly added to the concentrated red ginseng extract A and B, respectively, while stirring, until the alcohol concentration range is 50%, and placed in a 4°C refrigerated storage for 12h, using a Buchner funnel filter bottle to separate the precipitate and the supernatant of the alcohol precipitate, the precipitate is washed 5 times with 95% industrial ethanol until the supernatant is colorless, and the precipitate is dried under reduced pressure to obtain red ginseng polysaccharides A and B.

[0030] The molecular weight polysaccharides A and B are mixed in a ratio of 1:3 to prepare a red ginseng dendritic macromolecular polysaccharide concentrated solution with a concentration of 10 g / L, the solution pH is adjusted to 3 with a 10% hydrochloric acid solution, heated and boiled for 10 minutes, centrifuged at 3000 rpm for 10 minutes after cooling, and the supernatant is obtained. The supernatant is adjusted to pH 11 with a 10% sodium hydroxide solution, heated and boiled for 10 minutes, centrifuged at 3000 rpm for 10 minutes after cooling, and the supernatant is obtained. The supernatant is filtered using a 0.45 μm microporous filter membrane to obtain the pretreated red ginseng dendritic macromolecular polysaccharide solution.

[0031] Red ginseng and Ophiopogon japonicus were respectively added with 3BV 50% ethanol and refluxed at 90°C for 1h for extraction 5 times, the extracts were collected and the ethanol was recovered under reduced pressure until 0.3g of crude drug was contained in each 1mL, 1% activated carbon was added and stirred for 1h, the extracts were concentrated under reduced pressure until there was no alcohol taste, water for injection was added until 2.0g of crude drug was contained in each 1mL, and the alcohol extract concentrate of Shenmai injection (5 times concentration, 5 times concentration is relative to the concentration of finished Shenmai injection) was obtained after filtration and mixing, 200mL of Shenmai injection alcohol extract concentrate was taken, 200mL of pretreated 10g / L red ginseng dendritic macromolecular polysaccharide concentrated solution was added, 0.5% vitamin C was added as an antioxidant, and boiling and deoxygenated injection water was added to prepare 1000mL solution, 10% sodium hydroxide was used to adjust the pH to 8, and the solution was sealed in an ampoule under N2 protection, and the Shenmai injection in which the dendritic macromolecular polysaccharide replaced Tween 80 was obtained by dry heat sterilization at 121°C for 20min.

[0032] Application of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0033] Application of concentrated solution of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0034] Example 2

[0035] Licorice pieces were extracted three times with 3BV 70% ethanol at 80°C for 2h, and the obtained liquorice alcohol-extracted residues were dried at normal pressure in a drying oven at 50°C. The dried liquorice alcohol-extracted residues were then extracted three times with 10BV purified water, each time for 30min, to obtain a crude liquorice polysaccharide water extract, which was centrifuged at 1000rpm for 30min, and the supernatant was ultrafiltered using a hollow fiber column to prepare liquorice polysaccharides, and component A: 1KD~10KD and component B: 10KD~100KD molecular weight range were collected. The liquorice polysaccharide solution is concentrated under reduced pressure to a relative density of 1.30, the vacuum degree of reduced pressure concentration is 0 MPa, the concentration temperature is 60°C, 95% pure industrial ethanol is slowly added to the concentrated liquorice extract, stirring while adding until the alcohol concentration range is 90%, and the liquorice polysaccharide is stored in a refrigerator at 0°C for 6 hours, and a Buchner funnel filter bottle is used to separate the precipitate and the supernatant of the alcohol precipitate, the precipitate is washed 5 times with 95% industrial ethanol until the supernatant is colorless, and the precipitate is dried under reduced pressure to obtain liquorice polysaccharide.

[0036] The molecular weight polysaccharides A and B were mixed in a ratio of 1:3 to prepare a concentrated solution of 1 g / L of licorice dendrimer polysaccharide, the pH of the solution was adjusted to 4 using a 10% hydrochloric acid solution, heated and boiled for 30 minutes, centrifuged at 1000 rpm for 30 minutes after cooling, and the supernatant was obtained. The supernatant was adjusted to pH 14 using a 10% sodium hydroxide solution, heated and boiled for 20 minutes, centrifuged at 1000 rpm for 30 minutes after cooling, and the supernatant was filtered using a 0.45 μm microporous filter membrane to obtain a pretreated licorice dendrimer polysaccharide solution.

[0037] Red ginseng and Ophiopogon japonicus were respectively added with 3BV 70% ethanol and refluxed at 80°C for 2h for extraction 3 times. The extracts were collected and the ethanol was recovered under reduced pressure until 0.4g of crude drug was contained in 1mL. 1% activated carbon was added and stirred for 1h. The extracts were concentrated under reduced pressure until there was no alcohol taste. Water for injection was added until 2.0g of crude drug was contained in 1mL. After filtration and mixing, the alcohol extraction concentrate of Shenmai injection (5 times concentration) was obtained. 200mL of the concentrate was taken, 200mL of the pretreated 1g / L liquorice dendrimer polysaccharide solution was added, 0.5% sodium bisulfite was added as an antioxidant, and boiling and deoxygenated water for injection was added to prepare 1000mL of solution. 10% sodium hydroxide was used to adjust the pH to 7, and the extracts were sealed in ampoules under N2 protection. Licorice dendrimer polysaccharide replacing Tween 80 was obtained by dry heat sterilization at 115°C for 31min.

[0038] Application of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0039] Application of concentrated solution of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0040] Example 3

[0041] The epimedium pieces were extracted five times with 3BV 50% ethanol at 80°C for 2h under reflux, and the obtained epimedium alcohol-extracted drug residues were dried at normal pressure in a drying oven at 50°C. The dried epimedium alcohol-extracted drug residues were then extracted twice with 5BV purified water, each extraction time was 120min, and the crude epimedium polysaccharide water extract was obtained. The supernatant was taken and ultrafiltrated using a hollow fiber column to prepare epimedium polysaccharides, and component A: 1KD~10KD and component B: epimedium polysaccharides with a molecular weight range of 10KD~100KD were collected. The sugar solution is concentrated under reduced pressure to a relative density of 1.10, the vacuum degree of reduced pressure concentration is -0.1MPa, the concentration temperature is 90°C, 95% pure industrial ethanol is slowly added to the concentrated epimedium extract, stirring while adding, until the alcohol concentration range is 50%, and it is placed in a 4°C refrigerator for 12 hours, and a Buchner funnel filter bottle is used to separate the precipitate and the supernatant of the alcohol precipitate, and the precipitate is washed 5 times with 95% industrial ethanol until the supernatant is colorless, and the precipitate is dried under reduced pressure to obtain epimedium dendritic macromolecular polysaccharide.

[0042] The molecular weight polysaccharides A and B were mixed in a ratio of 1:3 to prepare a concentrated solution of epimedium polysaccharide with a concentration of 5 g / L, the solution pH was adjusted to 1 with a 10% hydrochloric acid solution, heated and boiled for 20 minutes, centrifuged at 12000 rpm for 5 minutes after cooling, and the supernatant was obtained. The supernatant was adjusted to pH 11 with a 10% sodium hydroxide solution, heated and boiled for 30 minutes, centrifuged at 12000 rpm for 5 minutes after cooling, and the supernatant was filtered with a 0.45 μm microporous filter membrane to obtain the pretreated epimedium dendritic macromolecular polysaccharide solution.

[0043] Red ginseng and Ophiopogon japonicus were respectively added with 3BV 50% ethanol and refluxed at 80°C for 2h for extraction 5 times. The extracts were collected and the ethanol was recovered under reduced pressure until 0.3g of crude drug was contained in each 1mL. 5% activated carbon was added and stirred for 0.5h. The extracts were concentrated under reduced pressure until there was no alcohol taste. Water for injection was added until 2.0g of crude drug was contained in each 1mL. After filtration and mixing, the alcohol extraction concentrate of Shenmai injection (5 times concentration) was obtained. 200mL of the concentrate was taken, 200mL of the pretreated 5g / L epimedium dendritic macromolecular polysaccharide concentrated solution was added, 0.1% sodium metabisulfite was added as an antioxidant, and 1000mL of injection water that had been boiled and deoxygenated was added to prepare a solution. 10% sodium hydroxide was used to adjust the pH to 6, and the solution was sealed in an ampoule under N2 protection. The solution was dry-heat sterilized at 120°C for 25min to obtain epimedium dendritic macromolecular polysaccharide replacing Tween 80 Shenmai injection.

[0044] Application of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0045] Application of concentrated solution of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0046] Example 4

[0047] Red ginseng slices were extracted 9 times with 10BV 90% ethanol at 70℃ for 5h, and the alcohol extract was used to prepare Shenmai injection. The obtained red ginseng alcohol-extracted residue was dried at normal pressure in a drying oven at 60℃. The dried red ginseng alcohol-extracted residue was then extracted twice with 5BV purified water, each time for 60min, to obtain a crude red ginseng polysaccharide water extract, which was centrifuged at 3000rpm for 10min, and the supernatant was ultrafiltered using a hollow fiber column to prepare red ginseng polysaccharides, and component A: 1KD~10KD and component B: 10KD~100KD molecules were collected. The red ginseng polysaccharide solution in the range of 100% was concentrated under reduced pressure to a relative density of 1.10, the vacuum degree of reduced pressure concentration was -0.1MPa, the concentration temperature was 90°C, 95% pure industrial ethanol was slowly added to the concentrated red ginseng extract, stirring while adding, until the alcohol concentration range was 50%, and it was placed in a refrigerated storage at 4°C for 12h, and the precipitate and the supernatant of the alcohol precipitate were separated by a Buchner funnel and suction flask, the precipitate was washed 5 times with 95% industrial ethanol until the supernatant was colorless, and the precipitate was dried under reduced pressure to obtain red ginseng polysaccharide.

[0048] The molecular weight polysaccharides A and B are mixed in a ratio of 1:3 to prepare a red ginseng polysaccharide concentrated solution with a concentration of 10 g / L, the solution pH is adjusted to 3 with a 10% hydrochloric acid solution, heated and boiled for 10 minutes, centrifuged at 3000 rpm for 10 minutes after cooling, and the supernatant is obtained. The supernatant is adjusted to pH 11 with a 10% sodium hydroxide solution, heated and boiled for 10 minutes, centrifuged at 3000 rpm for 10 minutes after cooling, and the supernatant is filtered with a 0.45 μm microporous filter membrane to obtain the pretreated red ginseng dendritic macromolecular polysaccharide solution.

[0049] Red ginseng and Ophiopogon japonicus were respectively added with 10BV90% ethanol and refluxed at 70°C for 5h for extraction 9 times, the extracts were collected and decompressed to recover ethanol until 0.4g of crude drug was contained in 1mL, 10% activated carbon was added and stirred for 3h, and the extracts were concentrated under reduced pressure until there was no alcohol taste, and water for injection was added until 2.0g of crude drug was contained in 1mL. After filtering and mixing, the alcohol extraction concentrate of Shenmai injection (5 times concentration) was obtained, 200mL of the concentrate was taken, 200mL of the pretreated 10g / L red ginseng dendritic macromolecular polysaccharide concentrated solution was added, 0.1% vitamin C was added as an antioxidant, and boiling and deoxygenated injection water was added to prepare 1000mL of solution, 10% sodium hydroxide was used to adjust the pH to 8, and the extracts were sealed in ampoules under N2 protection, and red ginseng dendritic macromolecular polysaccharide replacing Tween 80 Shenmai injection was obtained by dry heat sterilization at 121°C for 20min.

[0050] Application of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0051] Application of concentrated solution of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0052] Example 5

[0053] The red ginseng pieces were extracted 5 times with 3BV 70% ethanol at 90℃ for 1h, and the obtained red ginseng alcohol-extracted residues were dried at normal pressure in a drying oven at 50℃. The dried red ginseng alcohol-extracted residues were then extracted with 10BV 10% sodium hydroxide solution in cold water for 2 times, each extraction time was 60min, and the red ginseng crude polysaccharide water extract was obtained. The supernatant was centrifuged at 3000rpm for 10min, and the red ginseng dendritic macromolecular polysaccharide was prepared by hollow fiber column ultrafiltration, and component A: 1KD~10KD and component B: 10KD~100KD molecular weight were collected. The red ginseng polysaccharide solution in the range of 100% was concentrated under reduced pressure to a relative density of 1.10, the vacuum degree of reduced pressure concentration was -0.1MPa, the concentration temperature was 60°C, 95% pure industrial ethanol was slowly added to the concentrated red ginseng extract, stirring while adding, until the alcohol concentration range was 80%, and it was placed in a refrigerated storage at 4°C for 12h, and the precipitate and supernatant of the alcohol precipitate were separated by a Buchner funnel filter bottle, the precipitate was washed 3 times with 95% industrial ethanol until the supernatant was colorless, and the precipitate was dried under reduced pressure to obtain red ginseng dendritic macromolecular polysaccharide.

[0054] The molecular weight polysaccharides A and B were mixed in a ratio of 1:3 to prepare a red ginseng dendritic macromolecular polysaccharide concentrated solution with a concentration of 10 g / L, the solution pH was adjusted to 4 with a 10% hydrochloric acid solution, heated and boiled for 10 minutes, centrifuged at 3000 rpm for 10 minutes after cooling, and the supernatant was taken. The supernatant was adjusted to pH 9 with a 10% sodium hydroxide solution, heated and boiled for 10 minutes, centrifuged at 3000 rpm for 10 minutes after cooling, and the supernatant was filtered with a 0.45 μm microporous filter membrane to obtain the pretreated red ginseng polysaccharide solution.

[0055] Red ginseng and Ophiopogon japonicus were respectively added with 3BV 70% ethanol and refluxed at 90°C for 1h for extraction 5 times, the extracts were collected and the ethanol was recovered under reduced pressure until 0.3g of crude drug was contained in each 1mL, 1% activated carbon was added and stirred for 1h, and the extracts were concentrated under reduced pressure until there was no alcohol taste, and water for injection was added until 2.0g of crude drug was contained in each 1mL. After filtration and mixing, the alcohol extraction concentrate of Shenmai injection (5 times concentration) was obtained, 200mL of the concentrate was taken, 200mL of the pretreated 10g / L red ginseng polysaccharide concentrated solution was added, 0.5% sodium bisulfite was added as an antioxidant, and boiling and deoxygenated injection water was added to prepare a 1000mL solution, 10% sodium hydroxide was used to adjust the pH to 7, and the extracts were sealed in ampoules under N2 protection conditions, and red ginseng polysaccharide substituted Tween 80 Shenmai injection was obtained by dry heat sterilization at 121°C for 20min.

[0056] Application of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0057] Application of concentrated solution of dendritic macromolecular polysaccharide to replace Tween 80 in Shenmai injection.

[0058] Comparative Example 1

[0059] The sample preparation process is the same as that of Example 5, except that the alcohol-extracted residues of red ginseng are replaced with the alcohol-extracted residues of epimedium.

[0060] Comparative Example 2

[0061] The sample preparation process is the same as that in Example 5, except that the red ginseng dendritic macromolecular polysaccharide concentrated solution is replaced with a 25 mg / mL Tween 80 solution. The preparation method is as follows: 200 mL of the alcohol extract concentrate of Shenmai injection is taken, 200 mL of 25 mg / mL Tween 80 is added, 0.5% sodium bisulfite is added as an antioxidant, and 1000 mL of the solution is prepared by boiling and deoxygenating the injection water, 10% sodium hydroxide is used to adjust the pH to 7, and the solution is sealed in an ampoule under N2 protection, and dry-heat sterilized at 121°C for 20 min to obtain the red ginseng polysaccharide replacing Tween 80 Shenmai injection.

[0062] Comparative Example 3

[0063] The sample preparation process is the same as that of Example 5, except that no solubilizer is added, that is, the red ginseng dendritic macromolecular polysaccharide concentrated solution in Example 5 is omitted.

[0064] Comparative Example 4

[0065] The sample preparation process is the same as in Example 5, except that the red ginseng dendritic macromolecular polysaccharide is replaced with conventional water-extracted and alcohol-precipitated red ginseng polysaccharide.

[0066] The conventional preparation method of water-extracted and alcohol-precipitated red ginseng polysaccharide is as follows: take red ginseng slices and directly add 10BV purified water to heat and boil for 2 hours, extract twice in total, combine the extracts and concentrate under reduced pressure to a relative density of 1.10, add 95% ethanol to an alcohol concentration of 80%, precipitate and dry to obtain water-extracted and alcohol-precipitated red ginseng polysaccharide.

[0067] Comparative Example 5

[0068] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 1:1.

[0069] Comparative Example 6

[0070] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 3:1.

[0071] Comparative Example 7

[0072] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 1:4.

[0073] Comparative Example 8

[0074] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 1:1.

[0075] Comparative Example 9

[0076] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 3:1.

[0077] Comparative Example 10

[0078] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 1:3.

[0079] Comparative Example 11

[0080] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 1:1.

[0081] Comparative Example 12

[0082] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 3:1.

[0083] Comparative Example 13

[0084] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 10KD to 100KD, and the mass ratio of A:B is 1:3.

[0085] Comparative Example 14

[0086] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 1:1.

[0087] Comparative Example 15

[0088] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 3:1.

[0089] Comparative Example 16

[0090] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 1:3.

[0091] Comparative Example 17

[0092] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 1:1.

[0093] Comparative Example 18

[0094] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 3:1.

[0095] Comparative Example 19

[0096] The sample preparation process is the same as that of Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 1:3.

[0097] Comparative Example 20

[0098] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 1:1.

[0099] Comparative Example 21

[0100] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 3:1.

[0101] Comparative Example 22

[0102] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 30KD to 300KD, and the mass ratio of A:B is 1:3.

[0103] Comparative Example 23

[0104] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 1:1.

[0105] Comparative Example 24

[0106] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 3:1.

[0107] Comparative Example 25

[0108] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 1KD to 10KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 1:3.

[0109] Comparative Example 26

[0110] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 1:1.

[0111] Comparative Example 27

[0112] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 3:1.

[0113] Comparative Example 28

[0114] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 3KD to 30KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 1:3.

[0115] Comparative Example 29

[0116] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 1:1.

[0117] Comparative Example 30

[0118] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 3:1.

[0119] Comparative Example 31

[0120] The sample preparation process is the same as in Example 5, except that the molecular weight range of component A is 5KD to 50KD and the molecular weight range of component B is 50KD to 500KD, and the mass ratio of A:B is 1:3.

[0121] Test Trial 1:

[0122] The clarity of the injections obtained after sterilization in Example 5 and Comparative Examples 1 to 4 was observed, and foreign matter and insoluble particles were visible as shown in Table 1.

[0123] Compared with the traditional water-extracted and alcohol-precipitated red ginseng polysaccharides, red ginseng dendritic macromolecular polysaccharides have a more excellent solubilizing effect and can improve the stability of shengmai injection. This is due to the unique structure of red ginseng dendritic macromolecular polysaccharides extracted by the unique extraction method. Alcohol extraction of the decoction pieces will remove low molecular weight oligosaccharides and lipids in the decoction pieces, and improve the purity of the polysaccharides. Alkaline extraction will extract a large amount of high molecular weight acidic polysaccharides and reduce the extraction of starch-like polysaccharides. Combined with the ultrafiltration membrane with a porous structure, the spherical structure of the dendritic macromolecular polysaccharides can be more easily separated. The acidic dendritic macromolecular polysaccharides with higher purity will spontaneously aggregate in the solution to form a nanophase. The transmission electron microscopy of red ginseng dendritic macromolecular polysaccharides, such as Figure 1 As shown in the figure, the hydroxyl and carboxyl groups on the polysaccharide molecules easily form hydrogen bonds with various substances in the injection solution, thereby playing a solubilizing role in the form of nano-aggregates; the traditional water-extraction and alcohol-precipitation red ginseng polysaccharide extraction is too simple, the obtained red ginseng polysaccharide has low purity and contains more starch polysaccharides, and will not form a nano-phase state. The transmission electron microscopy of the water-extraction and alcohol-precipitation red ginseng polysaccharide is as follows Figure 2 As shown, the solubilization effect is poor and it is easy to precipitate in the solution system, affecting the stability of the injection system.

[0124] Table 1 Clarity of injection solutions with different solubilizers, visible foreign matter, insoluble particles

[0125]

[0126]

[0127] Test Trial 2:

[0128] The clarity of the injections obtained after sterilization in Examples 1 to 5 was observed, and foreign matter and insoluble particles were visible as shown in Table 2.

[0129] Table 2 Clarity of injection solutions with different solubilizers, visible foreign matter, insoluble particles

[0130]

[0131] Test Trial 3:

[0132] The clarity of the injection solutions obtained after sterilization of Comparative Examples 5 to 13 for 12 months is shown in Table 3.

[0133] Table 3 Clarity of injection solutions with different polysaccharide molecular weights and ratios

[0134]

[0135]

[0136] Test Trial 4:

[0137] The clarity of the injection solutions obtained after sterilization of Comparative Examples 14 to 22 for 12 months is shown in Table 4.

[0138] Table 4 Clarity of injection solutions with different polysaccharide molecular weights and ratios

[0139]

[0140]

[0141] Test Trial 5:

[0142] The clarity of the injections obtained after sterilization of Comparative Examples 23 to 31 for 12 months is shown in Table 5. By comparison, it is found that polysaccharides in the molecular weight range of 50KD to 500KD have a greater effect on clarity. This may be due to the large molecular weight of dendritic macromolecules. Dendritic macromolecular polysaccharides are more easily obtained in the ultrafiltration range of 50KD to 500KD. However, the concentration should be maintained within an appropriate range. Too high a concentration will still lead to poor stability of the injection.

[0143] Table 5 Clarity of injection solutions with different polysaccharide molecular weights and ratios

[0144]

[0145]

[0146] Test Trial 6:

[0147] According to Example 6 and Comparative Examples 2 and 3, Shenmai injection was prepared and placed in a stability test box for accelerated stability test at a temperature of 40±2°C and a relative humidity of 75%±5%. The clarity, pH, visible foreign matter, insoluble particles, and ginsenoside Rg1 content were tested at 0, 1, 2, 3, and 6 months, respectively. As shown in Table 6.

[0148] Table 6 Results of accelerated stability test of injection solutions with different solubilizers

[0149]

[0150]

[0151] The polysaccharide prepared by the present invention replaces Tween 80 injection and Shenmai injection, and the ingredients both contain ginsenoside Rg1. The accelerated test of Shenmai injection prepared in Example 5 and Comparative Examples 2 and 3 shows that the clarity, pH, visible foreign matter, insoluble particles, and ginsenoside Rg1 content of red ginseng polysaccharide Shenmai injection meet the requirements within 6 months, and the injection with red ginseng polysaccharide added is compared with the injection with Tween 80 added, and the clarity, pH, visible foreign matter, insoluble particles, and ginsenoside Rg1 content are not significantly different.

[0152] It can be seen from the above examples that the method provided by the present invention solves the problem of removing the stable vacancy of Tween 80 and overcomes the potential adverse effects caused by the addition of Tween 80.

[0153] It should be understood that in order to streamline the present disclosure and aid in understanding one or more of the various inventive aspects, in the above description of exemplary embodiments of the present invention, various features of the present invention are sometimes grouped together into a single embodiment, figure, or description thereof. However, this disclosed method should not be interpreted as reflecting the intention that the claimed invention requires more features than those expressly recited in each claim. Rather, as reflected in the claims, inventive aspects lie in less than all of the features of the previously disclosed embodiments. Therefore, the claims that follow the detailed description are hereby expressly incorporated into the detailed description, with each claim itself serving as a separate embodiment of the present invention.

[0154] Although the present invention has been described according to a limited number of embodiments, it will be apparent to those skilled in the art, with the benefit of the above description, that other embodiments may be envisioned within the scope of the invention thus described. In addition, it should be noted that the language used in this specification is selected primarily for readability and teaching purposes, rather than for explaining or defining the subject matter of the present invention. Therefore, many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the appended claims. The disclosure of the present invention is illustrative, not restrictive, with respect to the scope of the present invention, which is defined by the appended claims.

[0155] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine injection containing dendritic macromolecular polysaccharide, characterized in that: The Chinese medicine injection comprises 200 mL of the concentrated solution of the pretreated dendritic macromolecular polysaccharide, 200 mL of the injection alcohol extract concentrate, 0.1-0.5 wt% of an antioxidant, a pH regulator and water for injection, and the pH is adjusted to 6-8 to prepare a 1000 mL solution; The preparation method of the dendritic macromolecular polysaccharide comprises the following steps: using 3-10 BV 50-90% ethanol at 70-90°C to reflux extract the Chinese herbal medicine pieces for 3-9 times, each time for 1-5 hours, drying the obtained Chinese herbal medicine alcohol-extracted drug residues at normal pressure in a drying oven at 50-60°C, adding 5-10 BV purified water or 10% sodium hydroxide solution cold water to extract the dried Chinese herbal medicine alcohol-extracted drug residues for 2-3 times, each extraction time being 30-120 minutes, obtaining a Chinese herbal medicine crude polysaccharide water extract, centrifuging at 1000-12000 rpm for 5-30 minutes, taking the supernatant and using hollow fiber column ultrafiltration to prepare Chinese herbal medicine polysaccharides, collecting component A: a Chinese herbal medicine polysaccharide solution with a molecular weight range of 1KD-10KD and component B: a Chinese herbal medicine polysaccharide solution with a molecular weight range of 10KD-100KD, respectively, and reducing the pressure to concentrate component A and component B to a relative density of 1.10-1.30, and the vacuum degree of the reduced pressure concentration is -0.1-0 MPa, the concentration temperature is 60-90 ℃, 95% pure industrial ethanol is added to the concentrated Chinese medicine extract A and Chinese medicine extract B respectively, and stirred while adding until the alcohol concentration ranges from 50% to 90%, and placed in a 0-4 ℃ refrigerator for 6-12 h, and the precipitate and the supernatant of the alcohol precipitate are separated by suction filtration, and the precipitate is washed with 95% industrial ethanol until the supernatant is colorless, and the precipitate is dried under reduced pressure to obtain Chinese medicine polysaccharides A and Chinese medicine polysaccharides B, the molecular weight of Chinese medicine polysaccharide A is 1KD-10KD, and the molecular weight of Chinese medicine polysaccharide B is 10KD-100KD, and Chinese medicine polysaccharide A and Chinese medicine polysaccharide B are mixed in a mass ratio of 1:3 to obtain dendritic macromolecular polysaccharides; The method for preparing the concentrated solution of the pretreated dendritic macromolecular polysaccharide comprises the following steps: taking the dendritic macromolecular polysaccharide, preparing a concentrated solution of the dendritic macromolecular polysaccharide with a concentration of 1-10 g / L, adjusting the pH value of the solution to 1-4 with a hydrochloric acid solution, heating and boiling the solution for 10-30 minutes, cooling the solution and centrifuging the solution at 1000-12000 rpm for 5-30 minutes, taking the supernatant, adjusting the pH value of the supernatant to 9-14 with a sodium hydroxide solution, heating and boiling the solution for 10-30 minutes, cooling the solution and centrifuging the solution at 1000-12000 rpm for 5-30 minutes, taking the supernatant, and filtering the solution with a 0.45 μm microporous filter membrane to obtain the concentrated solution of the pretreated dendritic macromolecular polysaccharide; The Chinese herbal medicine slices include red ginseng slices, liquorice slices or epimedium slices; The preparation method of the alcohol extract concentrate for injection is as follows: taking red ginseng and ophiopogon japonicus, respectively adding 3-10BV 50-90% ethanol at 70-90°C to extract 3-9 times, each time for 1-5 hours, collecting the extract, recovering ethanol under reduced pressure until each 1mL contains 0.3-0.4g of the crude drug, adding 1-10% activated carbon and stirring for 0.5-3h, concentrating under reduced pressure until there is no alcohol taste, adding water for injection until each 1mL contains 2.0g of the crude drug, filtering and mixing to obtain the alcohol extract concentrate for ginseng and ophiopogon japonicus injection.

2. The Chinese medicine injection containing dendritic macromolecular polysaccharide according to claim 1, characterized in that: The antioxidant includes vitamin C, sodium bisulfite or sodium metabisulfite; the pH adjuster is 10% sodium hydroxide; and the water for injection is boiled and deoxygenated water for injection.

3. The method for preparing the Chinese medicine injection containing dendritic macromolecular polysaccharide according to any one of claims 1 to 2, characterized in that: The following steps are involved: Take 200 mL of the injection alcohol extract concentrate, add 200 mL of the pretreated concentrated solution of the dendritic macromolecular polysaccharide, add 0.1-0.5 wt % of an antioxidant, add boiling and deoxygenated injection water to prepare a 1000 mL solution, use a pH adjuster to adjust the pH to 6-8, fill in an ampoule under N2 protection, dry-heat sterilize at 115-121°C for 20-31 min, and obtain a traditional Chinese medicine injection containing dendritic macromolecular polysaccharide.

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