Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Diagnosis and treatment of cancer using anti-GPR49 antibody

An antibody and cancer technology, applied in the direction of antibodies, antibody mimetics/scaffolds, preparations for in vivo experiments, etc., can solve the problems such as hyperexpression of which are not seen

Inactive Publication Date: 2011-06-29
CHUGAI PHARMA CO LTD
View PDF32 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, it is also reported that: based on the expression of cancer cell lines, although GPR49 is overexpressed in colorectal cancer, ovarian cancer, glioma, and melanoma, it is not overexpressed in breast cancer and lung cancer (Non-Patent Document 8 )

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Diagnosis and treatment of cancer using anti-GPR49 antibody
  • Diagnosis and treatment of cancer using anti-GPR49 antibody
  • Diagnosis and treatment of cancer using anti-GPR49 antibody

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0265] (1) Preparation of effector cells

[0266] Spleens are removed from CBA / N mice and the like, and spleen cells are isolated in RPMI1640 medium (manufactured by Invitrogen). Wash with the same medium containing 10% fetal bovine serum (FBS, produced by HyClone), and then adjust the cell concentration to 5 × 10 6 cells / mL to prepare effector cells.

[0267] (2) Preparation of complement solution

[0268] A complement solution can be prepared by diluting Baby Rabbit Complement (manufactured by CEDARLANE) 10 times with a medium containing 10% FBS (manufactured by Invitrogen).

[0269] (3) Preparation of target cells

[0270] Cells expressing GPR49 protein were treated with 0.2mCi of 51 Cr-sodium chromate (manufactured by GE Healthcare Bio-Sciences) was incubated together in DMEM medium containing 10% FBS at 37° C. for 1 hour to radiolabel the target cells. As cells expressing GPR49 protein, cells transformed with a gene encoding GPR49 protein, gastric cancer, colon cance...

Embodiment 1

[0388] [Example 1] GPR49 mRNA expression analysis using human exon 1.0ST chip

[0389] In order to clarify the expression distribution of human GPR49 mRNA in clinical cancers, cancer cell lines, and various normal organs, expression analysis was performed using the human exon 1.0ST chip (Affymetrix) originally developed for splice variant analysis. The advantage of using the human exon 1.0ST chip for expression analysis is that compared with the existing expression chip of Affymetrix, which basically has only one probe set on the 3' side of each gene, the human exon 1.0ST At least one probe set can be set for each exon of the gene in the chip, so when using this chip to perform expression analysis on each gene, the expression data of multiple probe sets can be obtained for each gene. The reliability of expression data has been improved.

[0390] In this expression analysis, tumor sections from excised tissues of 22 lung adenocarcinomas, 13 tumors of gastric cancer, 5 tumors...

Embodiment 2

[0396] [Example 2] Establishment of cells expressing full-length human GPR49

[0397] Based on NCBI retrieval numbers NP_003658.1 (SEQ ID NO: 1 (amino acid sequence)) and NM_003667.2 (SEQ ID NO: 2 (nucleotide sequence)), the full-length human GPR49 cDNA was isolated by PCR, and This was cloned into a mammalian cell expression vector (pcDNA5 / FRT / TO) (Invitrogen). pcDNA5 / FRT / TO is a hybrid human CMV / TetO 2 The gene inserted under the promoter can be induced to express and is integrated with a neomycin resistance gene as a vector for drug resistance markers. Furthermore, the full-length human GPR49 cDNA was introduced into 293FlpIn T-Rex cells using the expression system FlpIn System (Invitrogen), which can induce expression only in the presence of tetracycline or doxycycline. In DMEM (high sugar) / 10%FBS / 100μg / mL Zeocin TM (Invitrogen) / 15 μg / ml blasticidin (Invitrogen) to introduce the expression vector into 293FlpIn T-Rex cells cultured with Fugene6 (Roche). According to t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
Login to View More

Abstract

Disclosed is an antibody which can bind to GPR49 protein and which has a cell proliferation-inhibiting activity against a cell capable of producing GPR49 protein. The cell proliferation-inhibiting activity may be a cytotoxic activity such as an antibody-dependent cytotoxicity or a complement-dependent cytotoxicity. Also disclosed are: a pharmaceutical composition; a cell proliferation inhibitor; and an anti-cancer agent, each of which comprises the antibody as an active ingredient. The cancer may be gastric cancer, colorectal cancer, hepatocellular cancer, lung cancer, prostate cancer, ovarian cancer, Ewing sarcoma, or glioma. Further disclosed are: a method for the diagnosis of cancer by detecting GPR49 protein or the expression of a gene encoding GPR49 protein; and a diagnosis agent and a kit for use in the method.

Description

technical field [0001] The present invention relates to a diagnosis method, a treatment method and an anticancer drug for cancer. Background technique [0002] The GPR49 molecule is a protein encoded by the ENSG00000139292 gene of human chromosome 12q12, and this molecule is a member of the LGR family (leucine-rich GPCR family, hereinafter referred to as the LGR family) of the hormone receptor family of G protein-coupled seven membrane-bound proteins. Member, which can be clarified by the characteristics of its amino acid sequence (Non-Patent Document 1). LGR family members include hormone receptors such as LHR, TSHR, and FSH, and LGR7, LGR8, etc., which use relaxin, insulin-like peptide 3 (INSL3), etc. as ligands (Non-Patent Document 2). All ligands are known to contain different peptides and mainly transmit signals mediated by cAMP. The LGR family has a structure comprising seven transmembrane domains and an N-terminal long extracellular domain in which there are 9-17 re...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30A61K39/395A61P35/00C07K19/00G01N33/536G01N33/574C12N15/09
CPCC07K2319/00C07K2317/565C07K2319/30C07K2317/24C07K2317/734G01N2333/726C07K2317/732C07K16/30G01N33/574A61P1/00A61P1/16A61P11/00A61P15/00A61P35/00A61P43/00C07K2317/34A61K51/1045G01N33/57492
Inventor 舟桥真一
Owner CHUGAI PHARMA CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com