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A kind of immunostimulatory compound and its preparation method and application

An immunostimulatory compound technology, applied in vaccines, drug combinations, pharmaceutical formulations, etc., can solve the problems of limiting the application of immune stimulating compound, no cost of reporting, scarcity of saponin resources, etc., to improve the immune function and survival rate of fish Improve and improve the effect of non-specific immune response

Active Publication Date: 2021-12-24
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the saponins used in the preparation of immunostimulatory complexes are mainly Quilaja saponins with the trade name QuilA, which are produced in South America and depend on imports. However, due to the scarcity of saponin resources, the cost of imports has increased year by year in recent years. Therefore, it is used to prepare immunostimulatory compounds. The cost of stimulatory complexes is also increasing, limiting the application of this immunostimulatory complex
The prior art does not report a low-cost active ingredient source that can replace QuilA

Method used

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  • A kind of immunostimulatory compound and its preparation method and application
  • A kind of immunostimulatory compound and its preparation method and application
  • A kind of immunostimulatory compound and its preparation method and application

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preparation example Construction

[0028] The present invention also provides a method for preparing the immunostimulatory complex described in the above technical scheme, comprising the following steps:

[0029] 1) using phosphate buffer as a solvent, mixing bovine serum albumin and surfactant, and standing at 37°C for 4 hours to obtain cleaved bovine serum albumin;

[0030] 2) mixing the lysed bovine serum albumin obtained in step 1) with Panax notoginseng saponins to obtain the first colorless transparent solution;

[0031] 3) mixing the first colorless and transparent solution obtained in step 2) with lecithin and cholesterol, and performing ultrasonic treatment to obtain a second colorless and transparent solution;

[0032] 4) The second colorless transparent solution obtained in step 3) was dialyzed in phosphate buffered saline at 4° C. for 72 hours to obtain an immunostimulatory complex.

[0033] The invention uses phosphate buffer saline as a solvent, mixes bovine serum albumin and surfactant, and stan...

Embodiment 1

[0041] Prepare 10ml of immunostimulatory complex

[0042] Bovine serum albumin (BSA) was dissolved in phosphate buffered saline (PBS, 0.01 mol / L, pH=7.4) to a solution with a protein concentration of 4 mg / mL. Cholesterol and lecithin were mixed and dissolved in chloroform in advance, and the concentrations were 250 mg / mL. Panax notoginseng saponins (PNS) were dissolved in PBS at a concentration of 1000 mg / mL. The preparation method is as follows: Take 8 mL of the above BSA solution and add 0.2 g of N-decanoyl-N-methylglucamine (Mega-10, product of Amresco Company) to stand in a 37°C incubator for 4 hours, then add 0.1 ml of the above PNS solution, Lecithin and cholesterol solution 10 μL, mix at room temperature, add PBS to a total solution volume of 10ml. The solution was placed in a mixture of ice and water at 0° C., and treated with ultrasonic waves (power 200 W) for 15 min. Finally, the solution was transferred into a dialysis bag, put into PBS solution and dialyzed at 4...

Embodiment 2

[0044] The structure of the immunostimulatory complex was observed by transmission electron microscopy.

[0045] Take an appropriate amount of the above-mentioned immunostimulatory complex, spot on the copper grid, use filter paper to absorb excess liquid from the edge after one minute, and then stain with acetic acid glaze, after one minute, use filter paper to absorb excess staining solution from the edge, and dry at room temperature , observed under a transmission electron microscope. The structure of the immunostimulatory complex observed under the transmission electron microscope is shown in figure 1 As shown, the spherical particle structure can be observed under the electron microscope, and the particle diameter is about 30nm. The particle is a ring structure formed by the combination of Panax notoginseng saponins and cholesterol, and is bonded into a spherical particle under the condition of the presence of lecithin.

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Abstract

The invention relates to an immunostimulatory compound and its preparation method and application, belonging to the technical field of immune preparations. The immunostimulatory complex provided by the present invention includes 4 mg / mL bovine serum albumin, 10 mg / mL Panax notoginseng saponins, 250 μg / mL cholesterol, 250 μg / mL lecithin and a surfactant with a mass concentration of 2%; The solvent is phosphate buffered saline. The immunostimulating compound provided by the invention has low cost and good immune effect on fish.

Description

technical field [0001] The invention relates to the technical field of immune preparations, in particular to an immune stimulating compound and its preparation method and application. Background technique [0002] The immunostimulatory complex is a spherical particle with a diameter of about 40nm formed by the hydrophobic interaction of saponins, lipids and antigen components. It can effectively present antigens to the immune system, improve the immune function of the body, and obtain better immune effects. At present, the saponins used in the preparation of immunostimulatory complexes are mainly Quilaja saponins with the trade name QuilA, which are produced in South America and depend on imports. However, due to the scarcity of saponin resources, the cost of imports has increased year by year in recent years. Therefore, it is used to prepare immunostimulatory compounds. The cost of stimulatory compounds is also increasing, limiting the application of this immunostimulatory...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61P37/04A61P31/04
CPCA61K39/39A61K2039/55577A61K2039/552
Inventor 陈秀霞龚晖许斌福池洪树陈永聪
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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