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Vaccine delivery vector as well as preparation method and application thereof

A delivery carrier and vaccine technology, applied in the field of vaccine delivery carrier and its preparation, can solve problems such as design difficulties and limited materials, and achieve the effects of improving delivery efficiency, reducing difficulty, and good specificity

Pending Publication Date: 2022-07-08
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the materials for efficient delivery of antigens are still very limited, and the development of self-adjuvant functional carriers often requires large-scale screening, the process is accompanied by chance, and the design is difficult. How to construct an efficient simplified tumor vaccine is still full of challenges

Method used

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  • Vaccine delivery vector as well as preparation method and application thereof
  • Vaccine delivery vector as well as preparation method and application thereof
  • Vaccine delivery vector as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Synthesis of PAMAM-R848 polymer

[0058] The immunoadjuvant-bonded polymer is obtained by Michael addition reaction between the amino group on the surface of the dendritic PAMAM polymer and the double bond of the R848 disulfide derivative.

[0059] In this embodiment, the dendritic PAMAM polymer is the fifth-generation PAMAM polymer, the molecular weight of the fifth-generation PAMAM polymer is 28826 g / mol, and 1 mol of the fifth-generation PAMAM contains 128 mol of amino groups. The synthetic route of PAMAM-R848 is as follows figure 1 shown.

[0060] The specific preparation steps of the immunoadjuvant-bonded polymer (ie PAMAM-R848) are as follows: 15 mg of fifth-generation PAMAM (G5-PAMAM) and 17.1 mg of HSEA-R848 are dissolved in 1.0 mL of anhydrous DMSO, HSEA-R848 and G5 - The molar ratio of PAMAM was 60:1, and the reaction was carried out in an oil bath at 40°C for 48 hours; the product was passed through a PD-10 gel column with ultrapure water as the ...

Embodiment 2

[0062] Example 2: PAMAM-R848 polymer and antigenic protein to prepare nanovaccine

[0063] The schematic diagram of the formation of nanovaccine between PAMAM-R848 polymer and protein antigen through electrostatic interaction and hydrophobic interaction is shown in Fig. image 3 shown in A.

[0064] In the verification experiment, a PAMAM-R848 aqueous solution with a concentration of 0.1-1 mg / mL was prepared, and then OVA model antigen was added at mass ratios of 2:1, 1:1, and 1:2 (polymer: antigenic protein), and vortexed. Mix well (the rotation speed is 500-2500rpm, just mix well) and incubate for 10 minutes to obtain the PAMAM-R848 / OVA (referred to as PRO) nano-vaccine. The PRO nanovaccine prepared with a mass ratio of 2:1 is denoted as PRO(2:1), and its dynamic light scattering (DLS) and transmission electron microscopy (TEM) characterization results are as follows image 3 As shown in B, the results show that the PRO(2:1) particle size is about 100 nm, and the morpholog...

Embodiment 3

[0066] Example 3: In vitro immune activation and antigen delivery effect of nanovaccine

[0067] As a TLR 7 / 8 agonist, R848 can promote the maturation of DC cells. In order to identify the immune activation function of PRO at the cellular level, the present invention selects the nanovaccine with three mass ratios obtained by the preparation method of Example 2: PRO (1:2 ), PRO(1:1), PRO(2:1). In the present invention, these three nano-vaccine and successfully induced BMDCs are incubated together for 24 hours, and the concentration of OVA is 10 μg / mL, and then CD11c, CD80 and CD86 antibodies are used to label the surface molecules of BMDCs, and flow cytometry is used to detect the expression level of costimulatory signal molecules , the streaming statistics are as follows Figure 4 shown in A. Compared with the control group (without vaccine and OVA) and the free OVA group (without immune adjuvant), the PRO vaccine can effectively promote the maturation of BMDCs in vitro, and...

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Abstract

The invention belongs to the technical field of biological medicine, and discloses a vaccine delivery carrier as well as a preparation method and application thereof. The vaccine delivery carrier is an immunologic adjuvant bonding polymer, and the structure of the vaccine delivery carrier is shown in the specification. The invention also discloses a preparation method of the vaccine delivery vector. The vaccine delivery vector provided by the invention can deliver a protein antigen to cytoplasm of an antigen presenting cell. According to the delivery carrier disclosed by the invention, the antigen cytoplasm delivery efficiency is improved, and the cross presentation of DC cell antigens is effectively promoted. The vaccine delivery carrier provided by the invention is used for preparing nano vaccines. The nano vaccine comprises a vaccine delivery carrier and a protein antigen. The vaccine provided by the invention is a vaccine for preventing tumors and / or resisting tumors. The vaccine inhibits tumor growth by inducing specific immune response, and is good in specificity and high in treatment efficiency.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, relates to the technical field of vaccines, and in particular relates to a vaccine delivery carrier and a preparation method and application thereof. Background technique [0002] Tumor immunotherapy kills tumor cells by regulating or stimulating the body's immune response. Compared with chemotherapy and radiotherapy, immunotherapy has stronger tumor specificity and less damage to normal tissues. Among the three immunotherapy methods, tumor vaccines simulate the process of the body being infected by pathogens to activate the immune response, and at the same time generate long-term immune protection through the immune memory function, which has been widely concerned. [0003] Tumor vaccines are usually composed of three parts: tumor antigen, immune adjuvant, and delivery carrier. The carrier protects the antigen from degradation and clearance and facilitates its delivery to lymph nodes (LNs) a...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K47/59A61K47/69A61K39/00A61P35/00
CPCA61K39/39A61K39/0011A61K47/595A61K47/6935A61P35/00A61K2039/55511A61K2039/876A61K2039/812A61K2300/00Y02A50/30
Inventor 杜金志徐炜高振林
Owner SOUTH CHINA UNIV OF TECH
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