Use of total saponin from Panax notoginseng stalks and leaves in preparing immunoadjuvants

An immune adjuvant and total saponin technology, which is applied in the direction of medical preparations containing active ingredients, antibody medical ingredients, pharmaceutical formulas, etc., can solve the problems of short immunity period, aluminum glue preparation process, carcinogen hazards, etc., and achieve enhanced immunity Originality, extensive regulation, wide range of pharmacological effects

Inactive Publication Date: 2012-07-25
秦枫
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But it has some insurmountable shortcomings: (1) No immunity to antigens of human immunodeficiency virus (HIV), hepatitis C virus (HCV), herpes simplex virus (HSV), influenza virus, schistosomiasis, whooping cough and injury Adjuvant effect; (2) aluminum glue preparation process, quality standard is difficult to standardize; (3) can only induce humoral immunity, can not induce cell-mediated immune response. Protozoa, etc.) and tumor immunity> is particularly necessary, far from meeting the development requirements of new vaccines
In the 1930s, the research on adjuvants reached its first climax, and the research on using white oil as an adjuvant became popular all over the world. However, what followed were strong side effects and the harm of carcinogens.
In the 1990s, although the new adjuvant represented by propolis can fully activate the cellular immune system and reduce stress, it has a short immune period, and it is prone to discoloration, precipitation, agglomeration, and side effects when stored at room temperature. a series of disadvantages
However, due to some unavoidable shortcomings such as different degrees of toxic side effects or safety hazards, their practical application is limited.

Method used

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  • Use of total saponin from Panax notoginseng stalks and leaves in preparing immunoadjuvants
  • Use of total saponin from Panax notoginseng stalks and leaves in preparing immunoadjuvants
  • Use of total saponin from Panax notoginseng stalks and leaves in preparing immunoadjuvants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Determination of saponin content in Panax notoginseng stem and leaf total saponins

[0019] (1) Preparation of reference substance and test solution solution Precisely weigh 2.48 mg of ginsenoside Rb1 reference substance dried under reduced pressure at 60°C to constant weight in a 25mL measuring bottle, add methanol to dissolve and dilute to the mark, shake well, and prepare Each mL contains 0.992mg of the reference substance solution. Accurately weigh 25.6 mg of total saponins in stems and leaves of Panax notoginseng in a 25 mL volumetric flask, add methanol to dissolve and dilute to the mark, shake well, and prepare a test solution containing 1.024 mg per mL.

[0020](2) Examination of linear relationship Precisely draw 0.5, 1, 1.5, 2, 2.5, 3.0mL of reference substance solution, place them in 10mL stoppered test tubes respectively, evaporate the solvent, and add newly prepared 5% vanillin-glacial acetic acid Solution 0.2mL, perchloric acid 0.8mL, heated in ...

Embodiment 2

[0028] Example 2 Hemolytic Determination

[0029] Blood was collected from the rabbit heart with a vacuum blood collection tube, placed in a triangular flask filled with glass beads, shaken gently for 15 minutes to remove fibrin, added twice the amount of normal saline for washing, centrifuged at 2000r / min for 10 minutes, washed 3 times, red blood cells The suspension was diluted to 2% with physiological saline. Take the above-mentioned total saponins from the stems and leaves of Panax notoginseng, filter it with a 0.45 μm microporous membrane, and dilute it with normal saline to make a 0.25mg / mL-5mg / mL dilution, and set up four repetitions, and the different concentrations of the test product Take 2 mL of each solution, add 2 mL of 2% erythrocyte suspension in the test tube, mix well, and use normal saline and distilled water as the maximum and minimum hemolysis controls respectively. Place the above-mentioned test tubes in a constant temperature water bath at 37°C for 1 hou...

Embodiment 3

[0035] Example 3 In vitro lymphocyte transformation test

[0036] Take 100 μL splenocyte suspension (1x10 7 each sample / mL) in a 96-well plate, each sample has 12 wells, of which 4 wells are added with ConA (final concentration 5 μg / mL), 4 wells are added with LPS (final concentration 10 μg / mL), 4 wells are added with 100 μL RPMI1640 solution, each well Add different concentrations of total saponin dilutions of Panax notoginseng stems and leaves (final concentrations are 0 μg / mL, 0.1 μg / mL, 1 μg / mL, 10 μg / mL, 100 μg / mL), and incubate for 48 hours at 37°C and 5% CO2. 4 hours before the end, 50 μL / well of MTT (2 mg / mL) was added to each well, and culture was continued for 4 hours. Discard the liquid in each well, add 150 μL of dimethyl sulfoxide to each well, place in a dark place at room temperature for 15 minutes, measure the OD value with a microplate reader at a wavelength of 578 nm, and see the results in figure 1 .

[0037] Depend on figure 1 It can be seen that the to...

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Abstract

The invention discloses the application of panax notoginseng stalk and leave total saponin in preparing immune adjuvant. The immune adjuvant prepared by adopting the panax notoginseng stalk and leave total saponin as a main effective component can remarkably strengthen the immunogenicity of antigens and can fulfill wide regulation action on the body immune function, and can induce immune responseto Th1 and Th2. The activity of the immune adjuvants is superior to aluminum hydroxide adhesive which is the only one immune adjuvant used by human up to now. The invention has low production cost and no toxicity.

Description

technical field [0001] The invention relates to the application of total saponins of notoginseng stems and leaves in the preparation of immune adjuvants. Background technique [0002] In recent years, with the in-depth study of basic immunology and the rapid development of antigen purification technology and genetic engineering technology, a variety of DNA recombinant vaccines and synthetic peptide vaccines have emerged. These new vaccines have high purity, small molecular weight, and specificity. Strong, good security and other advantages. But so far, very few related vaccines have been successfully developed. The main reason is that the purified antigen has weak immunogenicity, and it is difficult to induce an effective immune response in the body when used alone. An effective way to solve this critical problem is to use adjuvants. Since Glenny used aluminum hydroxide (alum) as an immune adjuvant in the 1920s, aluminum colloidal adjuvant is still the only adjuvant appro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39
Inventor 秦枫孙红祥葛竹兴朱善元郁杰方希修陈玉勇于生兰郑义张成洪伟鸣王妲妲
Owner 秦枫
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