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An affinity peptide ty peptide targeting dendritic cells and its application

A dendritic cell, targeting technology, applied in the field of tumor therapeutic proteins, can solve the problems of low antigen transport efficiency, weakening tumor effect, weak ability to stimulate anti-tumor immune response, etc.

Active Publication Date: 2021-04-27
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the preliminary application shows that the existing tumor vaccines have the defects of weak ability to stimulate anti-tumor immune response, the main reasons are low antigen transport efficiency, fast degradation rate in vivo, weak antigen cross-presentation ability, and most of them are delivered by non-professional antigens. The recognition of cells causes off-target effects of antigens, which in turn induces the incompetence of CTLs, thereby weakening the effect of treating tumors. Therefore, it is of great significance to screen new targeting DC molecules and design new tumor vaccines to improve the therapeutic effect of tumor vaccines.

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  • An affinity peptide ty peptide targeting dendritic cells and its application
  • An affinity peptide ty peptide targeting dendritic cells and its application
  • An affinity peptide ty peptide targeting dendritic cells and its application

Examples

Experimental program
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Embodiment 1

[0041] The affinity peptide TY peptide targeting dendritic cells provided in this application was screened by phage display peptide library technology, and the screening process included sorting mouse spleen DC cells, panning at the cell level, phage amplification, and affinity DC Steps such as determination of phage monoclonal DNA sequence, etc., this embodiment briefly introduces the specific screening process as follows.

[0042] (1) Sorting mouse spleen DC cells

[0043] Select 6-8 week old C57BL / 6J mice, kill and disinfect (soak in 75% alcohol for about 5 minutes), and prepare DC cells according to the existing technology, or refer to the following operations for details:

[0044] (1) After dissecting the mouse, take out the spleen and cut it into small pieces, add collagenase Ⅰ and DNA Ⅳ enzyme to digest the spleen cells, shake and digest at 37°C for 30 min;

[0045] (2) Grind the splenocytes with a glass slide, filter them with a 70-mesh filter to make a single cell su...

Embodiment 2

[0064] Based on the screening of TY peptide obtained in Example 1, the inventors synthesized DC's affinity peptide TY peptide using Fmoc solid-phase synthesis technology. It should be noted that when verifying the affinity of the synthesized TY peptide to DC, a method of indirectly reflecting the affinity of the peptide to DC was obtained by combining fluorescent streptavidin with biotin. After the TY peptide is synthesized, a biotin is further connected to the carboxyl terminal of the TY peptide through a linker-GGG-. In order to facilitate the preparation of subsequent nanocarriers, acetylated TY peptide and antigenic peptide OVA were simultaneously prepared. 257-264 . This embodiment briefly introduces the relevant preparation process as follows.

[0065] (1) Add the first amino acid

[0066] Weigh 0.37 g of Rink resin and pour it into the synthesizer, add DMF to make the resin swell for about 30 minutes (if Wang resin is used, do not deprotect); add the prepared deprote...

Embodiment 3

[0082] In the actual use of the screened TY peptide, it is limited by biological absorption and biodegradation. Therefore, this application uses the nanomaterial MSN (mesoporous silica nanoparticle) as a carrier to prepare a new type of nanoparticle that can target DC. Vaccine delivery system MSN-TY / OVA / CpG. During the preparation, the acetylated TY peptide was firstly covalently coupled to the surface of MSN by condensation reaction to synthesize MSN-TY, and then the mode antigen OVA and immune adjuvant CpG (CpG-ODN1826) were loaded by electrostatic adsorption.

[0083] The specific process of preparing MSN-TY (the connection of MSN-NH2 and COOH-peptide-Ac) is as follows:

[0084] Dissolve an appropriate amount of acetylated TY peptide in 2 mL of DMF, then add 10 times the equivalent of HoBt to mix and incubate, then add 10 times the equivalent of DIC, mix well with the above reactants, and incubate at room temperature for 10 min to activate the TY peptide. carboxyl;

[008...

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Abstract

This application belongs to the technical field related to protein for tumor therapy, and specifically relates to an affinity peptide TY peptide targeting dendritic cells and its application patent application matters. The TY peptide is a 12-peptide, and the specific amino acid sequence is: TITHFQFGPTVY. On the basis of TY peptide, this application further designed a new type of nano-transport system that can target DC based on MSN, and loaded the model antigen OVA and immune adjuvant CpG, and successfully prepared the nano-carrier MSN‑TY / OVA / CpG. Experiments show that MSN‑TY / OVA / CpG can be effectively taken up by immature DCs, improve the efficiency of antigen uptake by DCs, promote the maturation of DCs and effectively activate DCs, and effectively promote the processing and presentation of antigens , stimulate its secretion of cytokines, and further trigger antigen-specific CTL immune response and anti-tumor immune response.

Description

technical field [0001] This application belongs to the technical field related to protein for tumor therapy, and specifically relates to an affinity peptide TY peptide targeting dendritic cells and its application patent application matters. Background technique [0002] Tumors are a serious threat to human health. There are tens of millions of new cases and deaths every year in the world, of which my country accounts for about a quarter. Surgical treatment, radiotherapy and chemotherapy are routine methods for clinical treatment of tumors. Among them, the main problem faced by tumor treatment is late tumor metastasis and high recurrence rate; chemotherapy and radiotherapy have relatively high toxic and side effects, and the treated tumor is prone to develop resistance. Therefore, the development of highly efficient, specific, and low-toxic drugs is an urgent task for tumor treatment. [0003] Tumor immunotherapy makes full use of the activation mechanism of the body's own...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A61K9/14A61K47/64A61K47/52A61K39/39A61P35/00
CPCA61K9/143A61K9/146A61K39/39A61K2039/55561A61P35/00C07K7/08
Inventor 陈真真刘娅婷高艳锋姚琳通刘雅静
Owner ZHENGZHOU UNIV
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