Saponin with immunoadjuvant function, preparation method, vaccine preparation containing the saponin as adjuvant and uses thereof

A technology of immune adjuvant and effect, which is applied in the field of saponin with immune adjuvant effect and its preparation and vaccine preparation containing the saponin as adjuvant and application field, which can solve complex chemical structure, variability of aluminum glue, and strong hemolysis and other problems, to achieve the effect of simple preparation process, easy quality and convenient use

Inactive Publication Date: 2009-04-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are some problems with aluminum colloid adjuvant: ①Mainly through the induction of Th2 type immune response to produce antibodies to play a protective role, can not induce Th1 type immune response and cell-mediated immune response, only suitable for the main protective immunity of antibodies Vaccine; ②It has no adjuvant effect on various antigens such as human immunodeficiency virus, hepatitis C virus, herpes simplex virus, influenza virus, schistosomiasis, pertussis and typhoid fever; ③It can promote the production of IgE antibodies and easily induce allergic reactions in the body; ④ local reaction, granuloma will be formed, and local aseptic abscess may occur in very rare cases; ⑤ due to its physical and chemical properties, aluminum-containing glue vaccines are afraid of freezing, and aluminum glue is easy to denature after freezing; ⑥ may have harmful effects on the nervous system of humans and animals. influences
However, there are serious defects in Quil A: ① strong hemolysis, high toxicity and side effects, can cause local tissue necrosis, and even systemic adverse reactions and poisoning [Waite DC, Jacobson EW, Ennis FA, Edelman R, White B, Kammer R, et al .Three double-blind, randomized trials evaluating the safety and tolerance of different formulations of the saponinadjuvant QS-21.Vaccine 2001;19:3957-7.]
② Its main active ingredients such as QS-21, QS-17, QS-18 and QS-7 have acyl groups in their molecular structures, which are easy to hydrolyze, and their deacylated hydrolyzate loses the ability to induce Th1 type immune response and produce specific CTL ability, can only trigger Th2 type immune response [Kensil CR.Immunomodulatory adjuvants from Quillajasaponaria.Immunopotentiators in Modern Vaccines 2007; 109-22.]; Large, complex chemical structure, difficult to separate and prepare

Method used

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  • Saponin with immunoadjuvant function, preparation method, vaccine preparation containing the saponin as adjuvant and uses thereof
  • Saponin with immunoadjuvant function, preparation method, vaccine preparation containing the saponin as adjuvant and uses thereof
  • Saponin with immunoadjuvant function, preparation method, vaccine preparation containing the saponin as adjuvant and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Embodiment 1: preparation of platycodon saponin

[0037] Preparation of Platycodon grandiflora total saponins

[0038] Medicinal platycodon grandiflora coarse powder 5kg, reflux extraction with 70% ethanol for 3 times, 2 hours each time, filter, combine the filtrate, recover ethanol under reduced pressure, add appropriate amount of water to suspend the extract, extract with ether or petroleum ether until the ether liquid is nearly free color. The aqueous layer was extracted with water-saturated butanol or pentanol until the extract was nearly colorless, and the extracts were combined, recovered under reduced pressure and concentrated to obtain the alcohol extract. The alcohol extract is passed through a macroporous adsorption resin column, and the mixture of water and ethanol is used for gradient elution, and the eluent is checked by thin-layer chromatography. The fractions with the same spots are combined, concentrated, and separated to obtain the total saponins of Pl...

Embodiment 2

[0046] Embodiment 2 contains the hepatitis B vaccine preparation of saponin adjuvant

[0047] Weigh 500 μg of platycodon total saponins, platycodon saponin D or platycodon saponin D2, and 100 μg of hepatitis B virus surface antigen (HBsAg), dissolve in 10 ml of normal saline, filter through a 0.22 μm microporous membrane, and aseptically dispense, 1 ml each. Each ml contains saponin 50μg and HBsAg 10μg.

Embodiment 3

[0048] Example 3 Hepatitis B vaccine preparation containing aluminum gel and saponin adjuvant

[0049] Weigh an appropriate amount of platycodon total saponins, platycodon saponin D or platycodon saponin D2, dissolve in physiological saline to make a solution with a concentration of 100 μg / ml, and filter through a 0.22 μm microporous membrane. The saponin solution is mixed with an equal volume of aluminum-containing adjuvanted hepatitis B surface antigen vaccine stock solution (each ml contains 20 μg of HBsAg and 500 μg of aluminum hydroxide), mixed evenly, and aseptically dispensed, 1 ml per tube. Each ml contains saponin 50μg, HBsAg 10μg and aluminum hydroxide 250μg.

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PUM

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Abstract

The invention relates to a saponin with immune adjuvant function and a preparation method thereof, a vaccine preparation containing the saponin as an adjuvant, and applications of the saponin and the vaccine preparation in the prevention and treatment of infectious diseases and cancers of human and animals. The saponin is platycodin D, platycodin D2, or a total-saponin containing the two saponin compounds. The platycodin D and platycodin D2 are both extracted and separated from balloonflower, a Chinese medicine. The saponin can induce an organism to generate Th1-type and Th2-type immune responses, show the capability of inducing the organism to generate stronger cell immune response and humoral immune response to a vaccine than the alhydrogel adjuvant known in the prior art, and can be taken as the immune adjuvant for a plurality of vaccines and achieve an ideal immunity effect. The vaccine which takes the saponin as the adjuvant has simple preparation technology and simple and convenient method, and the quality is easy to control and the saponin can be reserved by freezing.

Description

technical field [0001] The present invention relates to saponin with immune adjuvant function, its preparation method, vaccine preparation containing the saponin as adjuvant, and their application in prevention and treatment of human and animal infectious diseases and cancer agents. Background technique [0002] Traditional vaccines are generally made of whole bacteria or whole viruses, and contain a large amount of non-immunogenic substances. These substances have certain adjuvant effects in addition to toxic and side effects, so generally do not need to add adjuvants [Lima KM, dos Santos SA , Rodrigues Jr JM, Silva CL. Vaccine adjuvant: it makes the difference. Vaccine 2004;22:2374-9]. With the continuous deepening of the basic theory of immunology and the rapid development of antigen purification technology and genetic engineering technology, many new vaccines such as gene vaccines, recombinant vaccines and synthetic peptide vaccines have emerged. These new vaccines have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07J63/00A61K39/39A61K36/34A61P31/00A61P35/00
Inventor 孙红祥胡松华
Owner ZHEJIANG UNIV
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