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Combined CTL antigenic epitope and its application

An antigenic epitope and immunogen technology, applied in the field of immunology, can solve the problems such as difficult breakthrough of CTL, achieve low maximum blood drug concentration, improve compliance, and improve the quality of life

Active Publication Date: 2013-08-21
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When a patient is immunized with a peptide vaccine alone, the peptide is easily degraded by peptidases in the body, and the induced CTL is not easy to break through the threshold required for tumor regression

Method used

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  • Combined CTL antigenic epitope and its application
  • Combined CTL antigenic epitope and its application
  • Combined CTL antigenic epitope and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 synthetic peptide

[0028] The peptide shown as SEQ ID NO: 4 was entrusted to Hangzhou Zhongpei Biochemical Co., Ltd. to synthesize it using a standard Fmoc protocol, and to use directional high-performance liquid chromatography for purification and purity analysis, and mass spectrometry for identification and molecular weight determination. The result shows that the purity of the polypeptide is higher than 95%, and the molecular weight is consistent with the theoretical value.

[0029] Specific synthesis steps: 1. Deprotection: The Fmoc-protected column and monomer must use a basic solvent, such as piperidine, to remove the amino protection group. 2. Activation and cross-linking: The carboxyl group of the next amino acid is activated and dissolved by the activator, and the activated monomer and the free amino group are cross-linked under the action of the cross-linking agent to form a peptide bond. 3. Cycle: The first two steps are repeated until the enti...

Embodiment 2

[0030] Example 2 Derivatives of Modified Polypeptides

[0031]Polyethylene glycol (PEG) is polymerized from ethylene oxide and consists of repeating oxyethylene groups. PEG not only has good water solubility, but also can be dissolved in organic solvents such as dichloromethane, N-N-dimethylacylphenol, benzene, acetonitrile and ethanol.

[0032] One end is blocked with a methyl group as methoxypolyethylene glycol (mPEG), and the molecular formula of linear mPEG is CH3-(O-CH 2 -CH 2 )n-OH, the derivatives of mPEG are most widely used in the study of PEGylation modification of polypeptides. Polyethylene glycol is a neutral, non-toxic high molecular polymer with unique physical and chemical properties and good biocompatibility. It is also one of the very few synthetic polymers approved by the FDA that can be used as a drug for intracorporeal injection. Polyethylene glycol is highly hydrophilic. When coupled to drug molecules, it can impart its excellent properties to the modif...

Embodiment 3

[0037] Example 3 Cytotoxic activity and the ability of vaccines to induce CTL-mediated cellular immunity in vitro

[0038] Effector cells: HLA-A * Anticoagulated fresh whole blood of 0201 positive healthy people was separated by conventional polysucrose-diatrizoate layered solution gradient centrifugation to obtain peripheral blood mononuclear cells, which were cultured with RPMI1640 containing 10% fetal bovine serum by volume solution to adjust the cell concentration to 1.0 x 10 6 / mL, inoculated into 24-well culture plate, 1 mL per well, each group stimulated PBMC 3 times continuously with epitopes with a concentration of 10 μg / mL and recombinant interleukin-2 (rhIL-2) with a concentration of 50 U / mL, weekly Once, the cells were collected on the 3rd day after the last stimulation to obtain epitope-induced specific CTLs, and the cell concentration was adjusted with RPMI1640 culture medium containing 10% fetal bovine serum by volume as effector cells.

[0039] 1 Multi-epitop...

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Abstract

The invention relates to the field of immunology, in particular to a combined antigenic epitope of tumor antigen, and also relates to application of the combined antigenic epitope. The combined CTL (cytotoxic T lymphocyte) antigenic epitope contains at least one polypeptide selected from SEQ ID NOS:1-3. The derivative is formed through modification of the epitope peptide by polyethylene glycol (PEG). After modification by PEG, the polypeptide has a longer half-life period, a low maximum plasma concentration, small plasma concentration fluctuation, less enzymolysis, less immunogenicity and antigenicity, less toxicity, better solubility, reduced medication frequency, and can improve patient compliance, improve the quality of life, and reduce the cost of treatment. A multi-epitope vaccine can stimulate human peripheral blood mononuclear T cells, can strongly induce the generation of antigen specific CTL, detects the ability of secreting functional cytokine IFN-gamma, and can cause specific dissolution breaking of a target cell in a target cell co-culture process. Thus, the multi-epitope vaccine is expected to become a vaccine for treatment of tumors.

Description

technical field [0001] The present invention relates to the field of immunology, in particular to combined antigenic epitopes of tumor antigens, and also relates to the application of combined antigenic epitopes. Background technique [0002] Lung cancer is one of the few malignant tumors that is the most common in the world and its incidence rate continues to increase. In western developed countries and my country, lung cancer has become the first cause of cancer death in men, and the death rate of lung cancer in American women has surpassed breast cancer in the late 1980s to become the first cause of cancer death in women. Therefore, lung cancer is the most serious malignant tumor threatening human health and life in the world. The morbidity and mortality of lung cancer in the Mainland of my country have also risen rapidly from the fourth and fifth places (male and female) in the 1960s to the first and second place today, especially in big cities and heavy industrial area...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/00A61P35/00
Inventor 吴玉章尚小云倪兵王莉曹毅杨玓李凡
Owner ARMY MEDICAL UNIV
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