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Preparation and application of cationic epitope vaccine

An epitope vaccine and cationic technology, which is applied in the field of self-assembly of cationic epitope peptides and anionic adjuvants to form cationic epitope vaccines, can solve autoimmune diseases that induce body inflammation, epitope vaccine toxicity and side effects, and no interaction and other issues, to achieve the effect of inhibiting tumor occurrence and growth, good biocompatibility, and eliminating toxic and side effects

Pending Publication Date: 2020-12-04
INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the addition of a large number of adjuvants can induce inflammation and autoimmune diseases in the body; and the addition of additional delivery systems often brings unpredictable toxic side effects to the application of epitope vaccines, such as inflammatory reactions, T cell inactivation, etc. Great obstacles to clinical application
Additionally, in these vaccines the antigen and adjuvant are simply physically mixed without interaction

Method used

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  • Preparation and application of cationic epitope vaccine
  • Preparation and application of cationic epitope vaccine
  • Preparation and application of cationic epitope vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Preparation of cationic epitope polypeptide

[0032] The epitope peptides described in the present invention are 5-20 amino acid antigen epitopes including T cell or B cell epitope polypeptides, specifically including but not limited to LWVFFDYVS, RWPSCQKKF, SIINFEKL, SVYDFFVWL, LCPGNKYEM, QAVHAAHAEINE, CYTWNQMNL, YMLDLQPETT, KSPWFTTL, SPSYVYHQF, EQLESIINFEKLTE, YEEYYPLI, EADPTGHSY, KIWEELSML, IMDQVPFSV, YMDGTMSQV, AAGIGILTV, and FLWGPRALI.

[0033] The cell penetrating peptides in the present invention include but not limited to

[0034] RRRRRRRR,RRRRRRRRR,RRRRRRRRRR,RRRRRRRRRRRR,RKKRRQRRR,LLIILRRRIRKQAHAHSK,GRKKRRQRRRPPQ,RKKRRRESRKKRRRES,KKKKKKKK,SQIKIWFQNKRAKIKK,GRPRESGKKRKRKRLKP,TRQARRNRRRRWRERQR,RRRRNRTRRNRRRVR,RKKRRRESRRARRSPRHL,SRRARRSPRHLGSG,GKRKKKGKLGKKRPRSR,SRRARRSPRESGKKRKRKR,LRRERQSRLRRERQSR,RCGRASRCRVRWMRRRRI。

[0035] Using SVYDFFVWL, LCPGNKYEM, and SIINFEKL as epitope polypeptides, the epitope polypeptides connected with polyarginine R8 gro...

Embodiment 2

[0043] Embodiment 2: Preparation of CpG / epitope nanocomposite vaccine

[0044] The cationized epitope polypeptide sequences SVY-R8, LCP-R8, SII-R8 and the anionic adjuvant CpG prepared in Example 1 were respectively dissolved in water to form a solution, and then respectively added to SII-R8, LCP-R8, Add CpG aqueous solution to the SVY-R8 aqueous solution at a ratio of 30:1 (w / w), mix with a vortex mixer, and incubate at 4°C for 20 minutes to obtain a self-assembled nano-epitope vaccine. The particle size of the nano-vaccine was detected by DLS, and its morphology was observed by TEM. The results are as follows: image 3 shown.

Embodiment 3

[0045] Example 3: Agarose Gel Electrophoresis of Epitope Peptide / CpG Nanocomplexes

[0046] The mass ratio of epitope peptide modified by R8 group to CpG was set as 10:1, 30:1, 50:1 and 60:1, and agarose gel electrophoresis experiment was carried out. Take the agarose solution in the Erlenmeyer flask

[0047] Preparation of agarose gel plate: add 1g of agarose to 50mL-1×TAE buffer solution, heat in microwave until the agarose is completely melted, and shake well to obtain 2.0% agarose gel. Add 1.5 μl ethidium bromide staining solution to the agarose solution and mix well. After a little cooling, carefully pour the agarose gel solution cooled to about 65°C into the glue tank, so that the glue solution slowly spreads until the entire glass plate A uniform gel layer is formed on the surface, and it is allowed to stand at room temperature until the gel is completely solidified, and the comb is pulled out vertically.

[0048] Sample application: Mix the nanocomposite aqueous solu...

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Abstract

The invention relates to a self-assembled nano epitope vaccine and application thereof. A T cell epitope peptide is subjected to cationization modification, so that the T cell epitope peptide and an anionic adjuvant are self-assembled in an aqueous solution to form a nano composite vaccine through electrostatic interaction. The vaccine can prevent or treat tumors or serve as a combined preparationof an anticancer active preparation. The epitope vaccine has the advantages of clear components, easiness in preparation, specificity cavity, high safety and good clinical transformation and practical application prospects. The problems of low synthetic efficiency, difficulty in separation and purification and the like of a synthetic vaccine are solved. Compared with other vaccines, the epitope vaccine has the advantages that a preparation method is simple, the size of formed vaccine nanoparticles is favorable, an additional delivery carrier is not needed, self-delivery of an antigen and theadjuvant is realized, activation of antigen presenting cells is promoted, an antitumor immune response is induced, tumor growth is inhibited, and tumor prevention and treatment functions can be realized.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and in particular relates to self-assembly of cationic epitope peptide and anionic adjuvant to form cationic epitope vaccine. Background technique [0002] Epitope peptide vaccine is a new type of vaccine prepared based on antigenic epitope. Compared with traditional vaccines, epitope peptide vaccine is safe, non-toxic, stable, and has low cost, high specificity, and adjustable immune type. High research and clinical application value. The epitope peptide vaccine directly delivers the most effective information fragment in the antigen—antigen epitope, which saves the complicated and inefficient antigen processing process, and can directly combine with MHC molecules to present antigen information, which can induce more effective and specific immune response. However, epitope vaccines are easily degraded in vivo, resulting in low immunogenicity, and the synthesis efficiency of chemically syn...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K39/385A61K39/00A61K47/64A61P35/00
CPCA61K39/39A61K39/385A61K39/0011A61K47/64A61P35/00A61K2039/55561A61K2039/5154A61K2039/5158A61K2039/6093
Inventor 王伟伟时圣彬宋会娟黄平升张闯年时小光
Owner INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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