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MAGE (Melanoma Antigen Gene)-4 anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide and application thereof

A MAGE-4, 1. MAGE-4 technology, applied in the field of biochemistry, can solve problems such as failure to achieve tumor inhibitory effect

Active Publication Date: 2012-06-27
SHANGHAI LONGYAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recent studies have shown that although the modified peptide vaccine based on the epitope MHC binding site can improve the immunogenicity of the vaccine to a certain extent, it cannot achieve the ideal anti-tumor effect in tumor immunotherapy; while the peptide vaccine based on the epitope TCR binding site The point-modified peptide vaccine is expected to achieve the ideal anti-tumor effect by enhancing the ability to stimulate low-affinity T cells or recruiting a new group of cross-reactive T cells to break tumor immune tolerance

Method used

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  • MAGE (Melanoma Antigen Gene)-4 anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide and application thereof
  • MAGE (Melanoma Antigen Gene)-4 anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide and application thereof
  • MAGE (Melanoma Antigen Gene)-4 anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of anti-tumor CTL epitope peptide P286 (Lys-Val-Leu-Glu-His-Val-Val-Arg-Val)

[0026] The Fmoc solid-phase synthesis method is used to extend one by one from the C-terminus to the N-terminus. Use fluorenylmethoxycarbonyl (Fmoc) to protect the α-amino group of amino acid, and the side chain protecting groups of various Fmoc protected amino acids are Arg (Pbf), His (trt), Glu (OtBu), Lys (Boc) (Pbf represents 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl, trt represents trityl, OtBu represents tert-butyl ester, Boc represents tert-butoxycarbonyl). First, use N, N'-diisopropylcarbodiimide (DIC) as the condensing agent for the first amino acid at the C-terminal of the α-amino protected C-terminal, and add 1-hydroxybenzotriazepam azole (HOBt), Fmoc-Val-OH is connected to the wang resin; sequentially washed with DMF, anhydrous methanol, dichloromethane, DMF; then piperidine-DMF mixture (V 哌啶 :V DMF =1:4) to remove the Fmoc protecting group, follo...

Embodiment 2

[0028] Example 2: Preparation of anti-tumor CTL epitope peptide P286-1Y2L (Tyr-Leu-Leu-Glu-His-Val-Val-Arg-Val)

[0029] The same synthesis method as in Example 1 was adopted, except that the second N-terminal amino acid Val was replaced by Leu, and Lys was replaced by Tyr.

[0030] The product was analyzed by mass spectrometry, see figure 2 , the result confirmed that the molecular weight was 1127.4, which was consistent with the theoretical value.

Embodiment 3

[0031] Example 3: Preparation of anti-tumor CTL epitope peptide P286-1Y2L9L (Tyr-Leu-Leu-Glu-His-Val-Val-Arg-Leu)

[0032] The same synthesis method as in Example 1 was adopted, except that the second amino acid Val at the N-terminus was replaced by Leu, the ninth amino acid Val at the N-terminus was replaced by Leu, and Lys was replaced by Tyr.

[0033] The product was analyzed by mass spectrometry, see image 3 , the results confirmed that the molecular weight was 1141.4, which was consistent with the theoretical value.

[0034] The CTL epitope peptide prepared above can be used to prepare a tumor therapeutic polypeptide vaccine, and its application experiment is as follows:

[0035] 1. Binding test of epitope peptide and HLA-A2.1 molecule

[0036] (1) Collect HLA-A2.1-positive T2 cells (ATCC company) with positive expression of HLA-A2.1 and lack of endogenous antigen processing ability by centrifugation at 800rpm, wash 3 times with phosphate buffered saline (PBS) at pH 7....

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Abstract

The invention relates to an MAGE (Melanoma Antigen Gene)-4 anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide. The epitope peptide is a nonapeptide, and the sequence of the nonapeptide is a-b-Leu-Glu-His-Val-Val-Arg-c, wherein a is Lys or Tyr, b is Val or Leu, and c is Val or Leu; the sequence of an epitope peptide P286 is Lys-Val-Leu-Glu-His-Val-Val-Arg-Val; the sequence of an anti-tumor CTL epitope peptide P286-1Y2L is Tyr-Leu-Leu-Glu-His-Val-Val-Arg-Val; and the sequence of an anti-tumor CTL epitope peptide P286-1Y2L9L is Tyr-Leu-Leu-Glu-His-Val-Val-Arg-Leu. The specific CTL induced in the body of a transgenic mouse by the anti-tumor CTL epitope peptide has certain kill rate to tumor cells EC-9706; and the epitope peptide can be used for preparing therapeutic tumor polypeptide vaccines.

Description

technical field [0001] The invention belongs to the technical field of polypeptides in the field of biochemistry, and in particular relates to an anti-tumor CTL epitope peptide derived from MAGE-4 and its application in preparing tumor therapeutic polypeptide vaccines. Background technique [0002] MAGE-4 belongs to the tumor-testis antigen (Cancer-Testis Antigen, CTA), and is one of the members of the MAGE gene family. According to reports, MAGE-4 is expressed in a variety of human malignant tumors, such as ovarian cancer, head and neck tumors, esophageal cancer, colon cancer, lung cancer, bladder cancer, etc., in esophageal squamous cell carcinoma, lung cancer, head and neck tumors, bladder cancer The expression levels in were particularly high, at 74%, 59%, 53% and 45%, respectively. Zambon et al found that 67% of esophageal squamous cell carcinoma and 37.5% of esophageal adenocarcinoma express at least one MAGE gene, and MAGE-4 is more common in squamous cell carcinoma ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K1/06C07K1/04A61K39/00A61P35/00
Inventor 祁元明吴宗胤高艳锋吕虹刘伟吴亚红李璐
Owner SHANGHAI LONGYAO BIOTECH CO LTD
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