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A gene vaccine carrier, its preparation method and application

A gene vaccine and carrier technology, applied in the field of its preparation, gene vaccine carrier, to achieve the effect of mild preparation conditions, simple process and strong immunogenicity

Active Publication Date: 2017-07-11
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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  • A gene vaccine carrier, its preparation method and application
  • A gene vaccine carrier, its preparation method and application
  • A gene vaccine carrier, its preparation method and application

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: 2mgNap-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: Dissolve 2mgNap-GFFY(p)-NHMe (prepared in Example 1) in 1mL PBS buffer, and configure into a 2‰ (w / v) small molecule peptide solution, with 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 carrier, its preparation method and application. The gene vaccine carrier is a supramolecular hydrogel formed by phosphatase-catalyzed small molecule polypeptide. The structural formula of the small molecule polypeptide is shown in formula (I): Wherein, when m=1, n=0, 1, 2 or 3; when n=1, m=1, 2 or 3. After the gene vaccine carrier is loaded with DNA, it has strong immunogenicity, large loading capacity, no obvious toxicity and can be injected for immunity. The preparation condition of the gene vaccine carrier of the invention is mild and the process is simple.

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 Patents(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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