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Tumor antigen peptide

a tumor antigen and peptide technology, applied in the field of detection agents, can solve the problems of insufficient therapeutic effect of anticancer agents that have been developed so far, inability of conventional therapeutic agents to selectively target cells that form the basis of cancer tissue, and low probability of curing cancer

Active Publication Date: 2018-05-31
ASSOC FOR KOKICHI KIKUCHI MEMORIAL ACADEMIC PROMOTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a new tumor antigen peptide that can help induce a specific CTL that targets cancer stem cells. This peptide can be used as an active ingredient in pharmaceutical compositions to prevent and treat cancer.

Problems solved by technology

The therapeutic effect of anticancer agents that have been developed so far is not sufficient and the probability of curing a cancer is very low.
As a cause thereof, the inability of conventional therapeutic agents to selectively target cells that form the basis of cancer tissue can be cited.
That is, the development of a technique for detecting cancer stem cells and a novel therapeutic agent that targets cancer stem cells are important issues in cancer medicine.

Method used

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  • Tumor antigen peptide
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Examples

Experimental program
Comparison scheme
Effect test

experimental example 1

ng of SP Fraction of Human Colon Cancer Cells

a) Preparation of Reagents

[0245]5% fetal calf serum (FCS (HyClone Laboratories))-supplemented DMEM (Sigma-Aldrich) medium was prepared as a medium and warmed at 37° C. Verapamil (Sigma-Aldrich) was adjusted to 50 mM and diluted to 5 mM using the 5% FCS-supplemented DMEM medium. Hoechst 33342 (Lonza) was adjusted to 250 μg / mL using the 5% FCS-supplemented DMEM medium. DNase I (Qiagen) was adjusted to 1 mg / mL using DDW and sterilized by filtration using a 0.2 μm filter.

b) Preparation of Cells for Flow Cytometry (FACS)

[0246]A human colon cancer cell line (SW480 (ATCC)) was suspended in 4 mL of the 5% FCS-supplemented DMEM medium, and the number of cells was counted. Furthermore, the 5% FCS-supplemented DMEM medium was added so as to adjust the cell concentration to 10×106 cells / mL, thus giving a sample. Using part of the sample, dispensing was carried out; verapamil was not added to a main sample (verapamil(−) sample), and verapamil was adde...

experimental example 2

ent

[0254]In order to confirm the in vivo tumorigenicity of each of the SW480-SP and SW480-MP clone cell lines obtained in Experimental Example 1, the SP clone and the MP clone were each transplanted to a NOD / SCID immunodeficient mouse (Oriental Kobo) using three representative clones thereof.

[0255]Specifically, the same number of SP and MP clone cells were suspended in 100 μL of 1×PBS on ice and mixed with 100 μL of Matrigel (BD). 100 μL of the cell Matrigel mixed solution was injected subcutaneously under the dorsal skin of a NOD / SCID mouse (Oriental Kobo) so as to give 100, 1000, and 10000 SP and MP clone cells for each group of five animals, and tumor development was examined. The major diameter and the minor diameter of a tumor were measured, and the tumor volume was calculated using the equation (volume=major diameter×(minor diameter)2 / 2). A tumor growth curve of a mouse into which 10000 cells had been transplanted is shown in FIG. 3.

[0256]From the results, in the 10000 cell-tr...

experimental example 4

oding HLA-A24-Binding Natural Peptide

a) SP-Specific Gene Expression

[0265]In Experimental Example 3e), a plurality of HLA-A24-binding natural peptides specific to the SP fraction cells were identified. These peptides are thought to be largely classified into two groups. They are a group for which a gene encoding the peptide is specifically expressed in the SP fraction cells and a group for which a gene encoding the peptide is expressed in both the SP fraction cells and the MP fraction cells, but due to differences in protein expression level or peptide processing, it is not subjected to antigen presentation by HLA-A24 as a natural peptide in the MP.

[0266]When, in order to classify the natural peptides identified above for the purpose of the above classification, mRNAs were extracted from SW480-SP and SW480-MP, and gene expression was examined by RT-PCR, the ASB4 gene was identified as one of genes specifically expressing in the SP fraction cell. The results of gene expression analysi...

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Abstract

The purpose of the present invention is to provide: a detection agent for specifically detecting a cancer stem cell; a tumor antigen peptide specifically presented by cancer stem cells; a medicinal composition useful in preventing and / or treating cancer, said medicinal composition comprising the aforementioned tumor antigen peptide as an active ingredient; a method for screening the tumor antigen peptide; etc. To achieve the above-mentioned purpose, provided are: peptides represented by YO-XO-ZO; a polyepitope peptide consisting of a plurality of epitope peptides connected together, said polyepitope peptide containing at least one of the above-mentioned peptides as one of the epitope peptides; a polynucleotide encoding the aforementioned peptides and / or polyepitope peptide; a medicinal composition comprising the same as an active ingredient; a prophylactic and / or therapeutic agent for cancer characterized by inducing CTL; etc.

Description

TECHNICAL FIELD[0001]The present invention relates to a detecting agent for detecting a cancer stem cell by using a gene that is specifically expressed in a cancer stem cell, a tumor antigen peptide derived from the gene, which is useful as a preventive and / or therapeutic agent for cancer, and the use thereof. Furthermore, the present invention relates to a method for screening such a tumor antigen peptide.BACKGROUND ART[0002]The therapeutic effect of anticancer agents that have been developed so far is not sufficient and the probability of curing a cancer is very low. As a cause thereof, the inability of conventional therapeutic agents to selectively target cells that form the basis of cancer tissue can be cited. In recent years, as such ‘cells forming the basis of cancer tissue’ the presence of cancer stem cells has been reported. Cancer stem cells are thought to be causal cells involved in the occurrence, recurrence, and metastasis of a cancer and, therefore, if cancer stem cells...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/00C12Q1/68C07K7/06C07K16/28C07K16/30C12N5/0783G01N33/50G01N33/574
CPCA61K39/0011C12Q1/6886C07K7/06C07K16/2833C07K16/30C12N5/0638G01N2333/46G01N33/574C12Q2600/136C12Q2600/158C12Q2600/178C07K2317/34A61K2039/572G01N33/5088A61K48/00C12Q1/68G01N33/5011G01N33/57484G01N2333/70539A61P35/00A61K2039/82A61K39/00118C07K14/47C07K14/4748A61K39/464838A61K39/4644A61K39/4611C07K7/08G01N2500/00
Inventor SATO, NORIYUKITORIGOE, TOSHIHIKOHIROHASHI, YOSHIHIKOKANASEKI, TAKAYUKIMIYAMOTO, SHOKOCHIN, VITALYGOTO, MASASHI
Owner ASSOC FOR KOKICHI KIKUCHI MEMORIAL ACADEMIC PROMOTION
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