Dendrobium huoshanense polysaccharide nanoemulsion immunologic adjuvant and preparation method thereof

Abstract

The invention discloses a dendrobium huoshanense polysaccharide nanoemulsion immunologic adjuvant and a preparation method thereof. The dendrobium huoshanense polysaccharide nanoemulsion immunologic adjuvant is prepared from the following raw materials in percentage by mass: 0.1%-2.0% of dendrobium huoshanense polysaccharide, 20%-30% of a surfactant, 5%-10% of a cosurfactant, 5%-10% of an oil phase and the balance of deionized water. The preparation method of the nano-emulsion is simple and easy to implement, equipment is easy to control, the cost is low, and the obtained nano-emulsion is small in droplet particle size, high in stability and capable of improving the spleen lymphocyte proliferation activity of immunocompromised mice. The nanoemulsion can significantly improve the immunomodulatory activity of dendrobium huoshanense polysaccharide, and has the potential of developing immunopotentiators, thereby promoting the development and utilization of traditional Chinese medicine resources and active components thereof.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical preparations, and particularly to a polysaccharide nanoemulsion immune adjuvant of Dendrobium huoshanense and a preparation method thereof. Background Art

[0002] Polysaccharide of Dendrobium huoshanense is the main active ingredient extracted from traditional Chinese medicine Dendrobium huoshanense. Research has found that polysaccharide of Dendrobium huoshanense has an improving effect on the reduction of phagocytic activity of mouse macrophages, the reduction of in vitro proliferation ability of splenic lymphocytes, and the reduction of thymus index induced by cyclophosphamide, and has a relieving effect on the hyperactive phagocytic activity of mouse peritoneal macrophages and the in vitro proliferation ability of splenic lymphocytes induced by Bacillus Calmette-Guérin. However, at present, in terms of application, the application form of polysaccharide of Dendrobium huoshanense is relatively single, mostly in traditional forms such as cut crude drugs and granules, and there are problems such as poor stability, poor solubility, less absorption, easy metabolism, and low bioavailability, and it cannot reach the level of finished drugs.

[0003] Nanoemulsion is a new type of drug delivery system. Applying nanoemulsion to the field of traditional Chinese medicine preparations can increase the solubility of poorly soluble drugs, and is also beneficial to the absorption of easily metabolized drugs in the body, effectively improving the bioavailability of traditional Chinese medicine components. Nanoemulsion also has sustained-release and controlled-release effects and targeting effects. By increasing the residence time of drugs in special parts of the body, the drug effect can be exerted continuously and slowly. However, there is currently no relevant research on nanoemulsion of polysaccharide of Dendrobium huoshanense. Summary of the Invention

[0004] Object of the Invention: To solve the problems existing in the prior art, the present invention provides a polysaccharide nanoemulsion immune adjuvant of Dendrobium huoshanense and a preparation method thereof. The obtained nanoemulsion has small particle size and strong stability, can improve drug solubility, promote drug absorption, and improve drug bioavailability.

[0005] Technical Solution: In order to achieve the above object of the invention, the technical solution adopted by the present invention is as follows:

[0006] A polysaccharide nanoemulsion immune adjuvant of Dendrobium huoshanense is composed of raw materials in the following mass percentages:

[0007]

[0008] Preferably, the surfactant is one or a mixture of several of Tween80, soybean lecithin, Span80, and Span85;

[0009] Preferably, the co-surfactant is one of propylene glycol, glycerol, and absolute ethanol.

[0010] Preferably, the oil phase is one of liquid paraffin, isopropyl myristate, and soybean oil.

[0011] Preferably, the particle size of the polysaccharide nanoemulsion immune adjuvant of Dendrobium huoshanense is 1-200 nm, and the dispersity is 0.01-0.3.

[0012] The present invention also provides a preparation method of the polysaccharide nanoemulsion immune adjuvant of Dendrobium huoshanense, comprising the following steps:

[0013] Dissolve the polysaccharide of Dendrobium huoshanense with deionized water to obtain an aqueous phase. After mixing the surfactant and the co-surfactant in a certain ratio, mix them with the oil phase in a certain ratio. After the solution is mixed evenly, slowly drop the aqueous phase into the mixture until a new equilibrium is reached to obtain a mixed solution; then add the remaining mass of deionized water and stir evenly to obtain a mixed solution. Ultrasonically treat the above mixed solution to obtain the polysaccharide nanoemulsion immune adjuvant of Dendrobium huoshanense.

[0014] Preferably, the ultrasonic power of the ultrasonic treatment is 700-900 W, and the ultrasonic frequency is 15-25 kHz.

[0015] Preferably, the ultrasonic time of the ultrasonic treatment is 5-10 min.

[0016] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0017] The present invention prepares the active ingredient polysaccharide of Dendrobium huoshanense into a nanoemulsion form. On the one hand, it is used to improve the bioavailability and drug effect of the polysaccharide of Dendrobium huoshanense and enhance the added value of Chinese medicinal materials produced in the Dabie Mountains. On the other hand, studying the application of nanoemulsions in the field of traditional Chinese medicine polysaccharide preparations can provide a reference for the dosage form improvement of other existing traditional Chinese medicine polysaccharide preparations and the improvement of the efficacy and clinical application value of active ingredients of traditional Chinese medicine.

[0018] The nanoemulsion preparation method of the present invention is simple and easy to implement, the equipment is easy to control, the cost is low, the obtained nanoemulsion droplets have a small particle size and strong stability, which can improve drug solubility, promote drug absorption, and improve drug bioavailability.

[0019] The polysaccharide nanoemulsion of Dendrobium huoshanense of the present invention can improve the proliferation activity of splenic lymphocytes in immunosuppressed mice and has the potential to develop immunopotentiators, thereby promoting the development and utilization of traditional Chinese medicine resources and their active ingredients. Specific embodiments

[0020] The following further illustrates the present invention through examples, but the following examples do not limit the scope of rights of this patent.

[0021] Example 1: Preparation of the polysaccharide nanoemulsion immune adjuvant of Dendrobium huoshanense

[0022] Take 0.1 g of polysaccharides from Dendrobium huoshanense, dissolve it in an appropriate amount of deionized water to obtain an aqueous phase. Weigh 20 g of Tween 80 and 5 g of anhydrous ethanol, mix them, then mix evenly with 10 g of soybean oil. Slowly add the aqueous phase dropwise to the mixture until a new equilibrium is reached to obtain a mixed solution. Then add the remaining mass of deionized water to make the total weight 100 g, and stir evenly to obtain a mixed solution. Place the mixed solution in an ultrasonic cell disruptor for ultrasonic treatment with an ultrasonic power of 800 W, an ultrasonic frequency of 20 kHz, and an ultrasonic time of 10 min to obtain the polysaccharides from Dendrobium huoshanense nanoemulsion immune adjuvant.

[0023] Example 2: Preparation of the polysaccharides from Dendrobium huoshanense nanoemulsion immune adjuvant

[0024] Take 0.5 g of polysaccharides from Dendrobium huoshanense, dissolve it in an appropriate amount of deionized water to obtain an aqueous phase. Weigh 20 g of soybean lecithin and 10 g of anhydrous ethanol, mix them, then mix evenly with 10 g of soybean oil. Slowly add the aqueous phase dropwise to the mixture until a new equilibrium is reached to obtain a mixed solution. Then add the remaining mass of deionized water to make the total weight 100 g, and stir evenly to obtain a mixed solution. Place the mixed solution in an ultrasonic cell disruptor for ultrasonic treatment with an ultrasonic power of 800 W, an ultrasonic frequency of 20 kHz, and an ultrasonic time of 10 min to obtain the polysaccharides from Dendrobium huoshanense nanoemulsion immune adjuvant.

[0025] Example 3: Preparation of the polysaccharides from Dendrobium huoshanense nanoemulsion immune adjuvant

[0026] Take 1 g of polysaccharides from Dendrobium huoshanense, dissolve it in an appropriate amount of deionized water to obtain an aqueous phase. Weigh 20 g of soybean lecithin and 10 g of propylene glycol, mix them, then mix evenly with 10 g of isopropyl myristate. Slowly add the aqueous phase dropwise to the mixture until a new equilibrium is reached to obtain a mixed solution. Then add the remaining mass of deionized water to make the total weight 100 g, and stir evenly to obtain a mixed solution. Place the mixed solution in an ultrasonic cell disruptor for ultrasonic treatment with an ultrasonic power of 800 W, an ultrasonic frequency of 20 kHz, and an ultrasonic time of 10 min to obtain the polysaccharides from Dendrobium huoshanense nanoemulsion immune adjuvant.

[0027] Example 4: Preparation of the polysaccharides from Dendrobium huoshanense nanoemulsion immune adjuvant

[0028] Take 2 g of polysaccharides from Dendrobium huoshanense, dissolve it in an appropriate amount of deionized water to obtain an aqueous phase. Weigh 30 g of soybean lecithin and 10 g of anhydrous ethanol, mix them, then mix evenly with 5 g of soybean oil. Slowly add the aqueous phase dropwise to the mixture until a new equilibrium is reached to obtain a mixed solution. Then add the remaining mass of deionized water to make the total weight 100 g, and stir evenly to obtain a mixed solution. Place the mixed solution in an ultrasonic cell disruptor for ultrasonic treatment with an ultrasonic power of 800 W, an ultrasonic frequency of 20 kHz, and an ultrasonic time of 10 min to obtain the polysaccharides from Dendrobium huoshanense nanoemulsion immune adjuvant.

[0029] Example 5: Preparation of Dendrobium huoshanense Polysaccharide Nanoemulsion Immunoadjuvant

[0030] Take 2 g of Dendrobium huoshanense polysaccharide and dissolve it in an appropriate amount of deionized water to obtain an aqueous phase. Weigh 20 g of Tween 80, 10 g of Span 85, and 10 g of absolute ethanol, mix them, and then mix them evenly with 5 g of soybean oil. Then, slowly drop the aqueous phase into the mixture until a new equilibrium is reached to obtain a mixed solution. Add the remaining mass of deionized water to make the total weight 100 g, and stir evenly to obtain a mixed solution. Place the mixed solution in an ultrasonic cell disruptor for ultrasonic treatment with an ultrasonic power of 800 W, an ultrasonic frequency of 20 kHz, and an ultrasonic time of 8 min to obtain the Dendrobium huoshanense polysaccharide nanoemulsion immunoadjuvant.

[0031] As mentioned above, it is only a general description of the present invention and does not impose any formal restrictions on the present invention. Any changes made by any other relevant person based on the relevant content of the present invention fall within the protection scope of the present invention.

Claims

1. A polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense, characterized in that, it is composed of raw materials in the following mass percentages:

2. The polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense according to claim 1, characterized in that, the surfactant is one or a mixture of several of Tween 80, soybean lecithin, Span 80, and Span 85.

3. The polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense according to claim 1, characterized in that, the co-surfactant is one of propylene glycol, glycerol, and absolute ethanol.

4. The polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense according to claim 1, characterized in that, the oil phase is one of liquid paraffin, isopropyl myristate, and soybean oil.

5. The polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense according to claim 1, characterized in that, the particle size of the polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense is 1 - 200 nm, and the dispersity is 0.01 - 0.

3.

6. The preparation method of the polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense according to any one of claims 1 - 5, characterized in that, it includes the following steps: Dissolve the polysaccharide from Dendrobium huoshanense with deionized water to obtain an aqueous phase. After mixing the surfactant and the co-surfactant in a certain proportion, then mix them with the oil phase in a certain proportion. After the solution is mixed evenly, slowly drop the aqueous phase into the mixture until a new equilibrium is reached to obtain a mixed solution; then add the remaining mass of deionized water and stir evenly to obtain a mixed solution. Ultrasonically treat the above mixed solution to obtain the polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense.

7. The preparation method of the polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense according to claim 6, characterized in that, the ultrasonic power of the ultrasonic treatment is 700 - 900 W, and the ultrasonic frequency is 15 - 25 kHz.

8. The preparation method of the polysaccharide nanoemulsion immune adjuvant from Dendrobium huoshanense according to claim 6, characterized in that, the ultrasonic time of the ultrasonic treatment is 5 - 10 min.

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