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Polymer coupled polypeptide nano vaccine and preparation method thereof

A nano-vaccine and polymer technology, applied in the direction of nano-technology, nano-technology, nano-drugs, etc., can solve the problems of reducing co-transportation efficiency, difficulty in determining the precise chemical composition of nano-vaccine, and weak non-covalent binding methods. Achieve the effect of stabilizing co-transported adjuvant molecules

Pending Publication Date: 2022-03-29
ZHEJIANG UNIV HANGZHOU GLOBAL SCI & TECH INNOVATION CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, on the one hand, this binding method has great restrictions on the type of adjuvant molecules (must be hydrophobic), and the non-covalent binding method is not strong, which may cause the adjuvant molecules to leak out, thereby reducing the co-transport efficiency
Therefore, the precise chemical composition of nanovaccine is also difficult to determine

Method used

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  • Polymer coupled polypeptide nano vaccine and preparation method thereof
  • Polymer coupled polypeptide nano vaccine and preparation method thereof
  • Polymer coupled polypeptide nano vaccine and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0058] Example 1, a preparation method of a polymer polypeptide-conjugated nano-vaccine. The molar ratio of polymer skeleton to grafted molecules is 1:8.1. Specifically prepared by the following steps:

[0059] 1) Preparation of mother solution: Dissolve 8ARM-PEG-mal in DMSO solution to obtain a PEG solution with a concentration of 5 mg / ml, labeled as S(PEG) solution; dissolve Pam adjuvant in DMSO solution to obtain a Pam solution with a concentration of 4 mg / ml, labeled For S (Pam) solution;

[0060] 2) Take 2 mg of G1 polypeptide, 220 μl of S(PEG) solution, 750 μl of DMSO solution, and 7 μl of TEA into a glass bottle of 1.5ml size, stir magnetically at a speed of 700 rpm, react at room temperature in the dark for 6 hours, and then add 100 mM cysteine The acid quenches the reaction.

[0061] 3) Dialyze the above mixed solution with a dialysis bag with a molecular weight cut off of 10000 Da to remove unreacted substances. Change 200ml of water every 4 hours, dialyze for a ...

Embodiment 2

[0062] Example 2, a preparation method of a polymer polypeptide-conjugated nano-vaccine.

[0063] The molar ratio of polymer skeleton to grafted molecules is 1:8.1.

[0064] Specifically prepared by the following steps:

[0065] 1) Preparation of mother solution: Dissolve 8ARM-PEG-mal in DMSO solution to obtain a PEG solution with a concentration of 5 mg / ml, which is marked as S(PEG) solution; dissolve Pam in DMSO solution to obtain a Pam solution with a concentration of 4 mg / ml, which is marked as S (Pam) solution;

[0066] 2) Take 2 mg of G1 polypeptide, 63 μl of S(Pam) solution, 438 μl of S(PEG) solution, 499 μl of DMSO solution, and 7 μl of TEA into a 1.5ml glass bottle, stir magnetically at 700 rpm, and react for 6 hours at room temperature in the dark , followed by the addition of 100 mM cysteine ​​to quench the reaction.

[0067] 3) Dialyze the above mixed solution with a dialysis bag with a molecular weight cut off of 10000 Da to remove unreacted substances. Change...

Embodiment 3

[0068] Example 3, a preparation method of a polymer polypeptide-conjugated nano-vaccine. Polymers 1:8.1. Concretely prepare through the following steps:

[0069] 1) Preparation of mother solution: Dissolve 8ARM-PEG-mal in DMSO solution to obtain a PEG solution with a concentration of 5 mg / ml, which is marked as S(PEG) solution; dissolve Pam in DMSO solution to obtain a Pam solution with a concentration of 4 mg / ml, which is marked as S (Pam) solution;

[0070] 2) Take 2 mg of G2 polypeptide, 204.5 μl of S (Pam) solution, 957 μl of S (PEG) solution, and 8 μl of TEA into a 1.5ml glass bottle, stir magnetically at 700 rpm, react at room temperature for 6 hours in the dark, and then add The reaction was quenched with 100 mM cysteine.

[0071] 3) Dialyze the above mixed solution with a dialysis bag with a molecular weight cut off of 10000 Da to remove unreacted substances. Change 200ml of water every 4 hours, dialyze for a total of 16 hours, then collect the obtained sample solu...

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Abstract

The invention discloses a polymer coupled polypeptide nano vaccine and a preparation method thereof. The vaccine comprises the following components in molar ratio: 1, a polymer skeleton with multiple reaction centers, 8.1, grafted molecules, 49167-213900, an organic solvent and 170-657 of organic alkali, the particle size of the nano vaccine can be controlled by regulating and controlling the stoichiometric ratio of the polypeptide to the Pam adjuvant molecule; the polypeptide nano vaccine obtained by the invention has a stable structure, is hardly influenced by concentration, has no concentration limiting condition, and has the capability of stabilizing co-transport adjuvant molecules.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a polymer-coupled polypeptide nano-vaccine and a preparation method thereof. Background technique [0002] The development and application of individualized neoantigen cancer vaccines has become a new way of cancer immunotherapy. However, due to the single naked peptide vaccine can only produce low immunogenicity. This is because naked peptides are easily decomposed and cleared in vivo, and the efficiency of phagocytosis and presentation by antigen-presenting cells (APCs), such as dendritic cells (DCs), is low. Due to the diversity of the nature of the polypeptide itself, through the design strategy of amphiphilic polypeptides, the structural characteristics of hydrophobic and hydrophilic segments can be endowed to the polypeptide molecule at the same time. This amphiphilic peptide can self-assemble into a supramolecular micelle structure in aqueous solution, but the premise of formi...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61K39/00A61K39/39A61P35/00B82Y5/00B82Y30/00B82Y40/00
CPCA61K39/385A61K39/0011A61K39/39A61P35/00B82Y5/00B82Y30/00B82Y40/00A61K2039/55516A61K2039/6093
Inventor 陈枢青王洪亮
Owner ZHEJIANG UNIV HANGZHOU GLOBAL SCI & TECH INNOVATION CENT
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