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Universal CpGODN nanoparticle adjuvant as well as preparation method and application thereof

A nanoparticle and adjuvant technology, which is applied in the field of universal CpGODN nanoparticle adjuvant and its preparation, can solve the problems of increasing CpGODN immune effect, CpGODN toxicity and side effects, limiting the use range of CpGODN immune adjuvant, body toxicity and side effects, etc., to achieve Good immune effect, improved utilization and safety, and reduced systemic inflammation

Inactive Publication Date: 2020-12-18
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, it is also necessary to increase the injection dose of CpG ODN in order to achieve the expected immune effect, but the increase in the injection dose will also cause the risk of systemic inflammation and produce toxic side effects on the body, which poses a safety risk
At present, the CpG ODN immune adjuvant and immune antigen are usually prepared together to form a nanoparticle vaccine. For example, the patent: CN 106421770A discloses a nanoparticle containing the EV71VP1 protein. The EV71 VP1 protein, immune adjuvant, cationic polymer and Anionic polymers are prepared into nanoparticles, although it can increase the immune effect of CpG ODN and reduce the toxic and side effects of CpG ODN to a certain extent, but it also limits the scope of use of CpG ODN immune adjuvants

Method used

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  • Universal CpGODN nanoparticle adjuvant as well as preparation method and application thereof
  • Universal CpGODN nanoparticle adjuvant as well as preparation method and application thereof
  • Universal CpGODN nanoparticle adjuvant as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Preparation method of nanoparticle adjuvant comprising CpG ODN

[0048] 1. Method

[0049] 1. Solution preparation

[0050] (1) Disperse protamine in sterilized distilled water, stir overnight under magnetic stirring, and filter through filter paper to obtain a protamine solution. (2) Disperse sodium tripolyphosphate in sterilized distilled water, stir for 5 minutes under magnetic stirring, and filter through a 0.22 μm filter membrane to obtain sodium tripolyphosphate solution. (3) Dissolve CpG in distilled water, stir for 5 minutes under magnetic stirring, and filter through a 0.22 μm filter membrane to obtain a CpG solution. (4) Using the method of fast nanocomplexation (FNC) (such as figure 1 ), channel 1: protamine solution; channel 2: sodium tripolyphosphate solution; channel 3: CpG-containing solution; channel 4: pure water, the flow rate of the syringe pump is 10 mL / min, and solution A can be obtained. (5) The concentration of protamine in step (1) ...

Embodiment 2

[0061] Example 2 Preparation method of nanoparticle adjuvant comprising CpG ODN

[0062] 1. Method

[0063] 1. Solution preparation

[0064] (1) Disperse protamine in sterilized distilled water, stir overnight under magnetic stirring, and filter through filter paper to obtain a protamine solution. (2) Disperse sodium tripolyphosphate in sterilized distilled water, stir for 5 minutes under magnetic stirring, and filter through a 0.22 μm filter membrane to obtain sodium tripolyphosphate solution. (3) Dissolve CpG in distilled water, stir for 5 minutes under magnetic stirring, and filter through a 0.22 μm filter membrane to obtain a CpG solution. (4) Using the method of fast nanocomplexation (FNC) (such as figure 1 ), channel 1: protamine solution; channel 2: sodium tripolyphosphate solution; channel 3: CpG-containing solution; channel 4: pure water, and the flow rate of the syringe pump was 10 mL / min to obtain solution B. (5) The concentration of protamine in step (1) is 0.5...

Embodiment 3

[0075] Example 3 Preparation method of nanoparticle adjuvant comprising CpG ODN

[0076] 1. Method

[0077] 1. Solution preparation

[0078] (1) Disperse protamine in sterilized distilled water, stir overnight under magnetic stirring, and filter through filter paper to obtain a protamine solution. (2) Disperse sodium tripolyphosphate in sterilized distilled water, stir for 5 minutes under magnetic stirring, and filter through a 0.22 μm filter membrane to obtain sodium tripolyphosphate solution. (3) Dissolve CpG in distilled water, stir for 5 minutes under magnetic stirring, and filter through a 0.22 μm filter membrane to obtain a CpG solution. 4. Using fast nanocomplexation (FNC) methods (such as figure 1 ), channel 1: protamine solution; channel 2: sodium tripolyphosphate solution; channel 3: CpG-containing solution; channel 4: pure water, the flow rate of the syringe pump is 10 mL / min, and solution C can be obtained. (5) The concentration of protamine in step (1) is 0.5 ...

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Abstract

The invention discloses a CpG ODN nanoparticle adjuvant as well as a preparation method and application thereof. The CpG ODN nanoparticle adjuvant comprises a CpG ODN immunologic adjuvant, a cationicpolymer and an anionic polymer. According to the CpG ODN nanoparticle adjuvant and the preparation method thereof, on one hand, controllable preparation of the nano-particle adjuvant can be achieved,a series of nano-adjuvants of different sizes can be prepared, and on the other hand, the metabolic rate of CpG ODN can be improved, and the residence time of CpG ODN in lymph nodes can be prolonged;the distribution of the CpG ODN adjuvant in vivo can be regulated, so that systemic inflammation is reduced, and the availability and the safety of CpG ODN are improved; the CpG ODN nanoparticles havesafety and universality, can be matched with different antigens for use, can be prepared into different vaccine according to requirements, and are simple and convenient.

Description

technical field [0001] The present invention relates to the technical field of nano adjuvant, more specifically, relates to a universal CpG ODN nano particle adjuvant and its preparation method and application. Background technique [0002] Immune adjuvants, also known as immune stimulants, refer to substances that can enhance the body's immune response to antigens or change the type of immune response of the body, thereby allowing the body to produce an effective immune response, and play an important role in vaccines. CpG ODN (CpG oligonucleotide, CpG oligodeoxynucleotide) is an oligodeoxynucleotide (ODN) containing unmethylated cytosine guanine dinucleotide (CpG) with immunostimulatory effect. [0003] Usually, each DNA single strand of CpG ODN has 2-4 CpG dinucleotides, and each backbone has two phosphorylation modification methods: phosphorothioate backbone (PTO) or phosphodiester backbone (PO). CpG ODN can be divided into 3 types according to its structure and activat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K9/51A61K47/42A61K47/30A61K47/34A61K47/36A61K39/145A61K39/00A61P31/16A61P35/00
CPCA61K9/5115A61K9/5146A61K9/5161A61K9/5169A61K39/0011A61K39/12A61K39/39A61K2039/55561A61P31/16A61P35/00A61K2039/876C12N2760/16134
Inventor 刘利新乔冬冬陈永明
Owner SUN YAT SEN UNIV
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