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Gene vaccine vector, and preparation method and application thereof

A gene vaccine and carrier technology, applied in the field of gene vaccine carrier and its preparation, achieves the effects of mild preparation conditions, large loading capacity and simple process

Active Publication Date: 2015-01-14
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The Chinese patent application (application number: 200910229186.7) involves a series of synthetic methods of polypeptide derivatives used to generate stable small molecule hydrogels, but does not involve the application of such small molecule hydrogels

Method used

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  • Gene vaccine vector, and preparation method and application thereof
  • Gene vaccine vector, and preparation method and application thereof
  • Gene vaccine vector, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Synthesis of Small Molecular Peptides

[0037] The synthetic method step of small molecule polypeptide used in the present invention is as follows:

[0038]

[0039] Among them, Fmoc-OSu is fluorenylmethoxycarbonyl succinimide, DIPEA is N,N-diisopropylethylamine, Acetone is acetone, I 2 is iodine, triethyl phosphite is triethyl phosphite, DCM is dichloromethane, pyridine is pyridine, TFA is trifluoroacetic acid, spps is the English abbreviation of "polypeptide solid-phase synthesis", TMSBr is trimethylbromosilane, and MeOH is Methanol, methylamine hydrochloride is methylamine hydrochloride.

[0040] Step 1, Fmoc-L-Tyr-O t Synthesis of Bu

[0041] Dissolve 10mmol L-tert-butyl tyrosine and 10 molecules of N,N-diisopropylethylamine in 75mL of acetone, add 9.8mmol of Fmoc-OSu containing 75mL of acetone solution under stirring , stirred at room temperature for 12 hours, and then separated by silica gel column to obtain 4.4 g of product Fmoc-L-Tyr-O t Bu (i...

Embodiment 2

[0059] Example 2: Preparation of supramolecular hydrogel (gene vaccine carrier)

[0060] Configuration solution 1: alkaline phosphatase 2U / μL, the buffer is 50mM Tris-HCl+1mM MgCl 2 ; Solution 2: 2mg Nap-GFFY(p)-NHMe (prepared in Example 1) was dissolved in 1mL PBS buffer, configured as a 2‰ (w / v) small molecule peptide solution, and Nap 2 CO 3 Adjust the pH to 9. Then, 3 μL of solution 1 was added to 100 μL of solution 2, mixed evenly, and allowed to stand at room temperature for at least 10 min to form a supramolecular hydrogel.

[0061] The prepared supramolecular hydrogel was observed by transmission electron microscope, such as figure 1 As shown, the prepared supramolecular hydrogel is a left-handed helix, which can compress DNA, protect its loaded DNA from degradation, and promote cell antigen expression.

Embodiment 3

[0062] Example 3: Preparation of supramolecular hydrogel loaded with HIV DNA (gene vaccine carrier loaded with DNA)

[0063] Solution 1: Alkaline phosphatase 2U / μL, the buffer is 50mM Tris-HCl, 1mM MgCl2; Solution 2: 2mg Nap-GFFY(p)-NHMe (prepared in Example 1) was dissolved in 1mL PBS buffer, Configured as a 2‰ (w / v) small molecule peptide solution, use Na 2 CO 3 Adjust the pH to 9, and add 10 μg of HIV DNA vaccine (pDRVI SV1.0-Env). Then, 3 μL of solution 1 was added to 100 μL of solution 2, mixed evenly, and allowed to stand at room temperature for at least 10 min to form a supramolecular hydrogel.

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Abstract

The invention discloses a gene vaccine vector, and a preparation method and an application thereof; the gene vaccine vector is a supramolecular hydrogel formed through phosphatase catalysis of a small-molecular peptide; the small-molecular peptide has the structural formula represented by the formula (I), wherein when m=1, n=0, 1, 2 or 3; when n=1, m=1, 2 or 3. The gene vaccine vector after being loaded with DNA has the advantages of strong immunogenicity, large load capacity, no obvious toxicity and injectable immunity. The gene vaccine vector is mild in preparation conditions and simple in process.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a gene vaccine carrier, its preparation method and application. Background technique [0002] Human immunodeficiency virus (Human Immunodeficiency Virus, HIV) has caused more than 30 million deaths, and it is urgent to develop a highly effective vaccine to prevent HIV infection. In addition, traditional vaccines such as hepatitis B, hepatitis C, and influenza have low efficacy and potential risks, so safe alternatives such as genetic vaccines have attracted widespread attention. Genetic vaccines are very safe and can produce long-term cellular and humoral immune responses, but the immunogenicity of genetic vaccines is very low, and they need to be delivered to the body through a carrier to enhance the immune response. Existing carriers include polymers, liposomes, and nano or micro particles. Polymers such as polyetherimide (PEI) have greater toxicity (Lim, M.H., et al. (2007). Accou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/24A61K39/29A61K39/21A61K39/145A61K9/06A61P31/20A61P31/14A61P31/18
Inventor 蒋兴宇田月杨志谋王怀民邵一鸣
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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