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Fibroblast growth factor receptor 2-specific peptide reagents and methods

A growth factor receptor and fibroblast technology, applied in the field of fibroblast growth factor receptor 2-specific peptide reagents, can solve the problems of no obvious advantages and low contrast.

Inactive Publication Date: 2020-05-05
RGT UNIV OF MICHIGAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Other methods of wide-field endoscopy, including color endoscopy 35 , Narrow Band Imaging (NBI) 36 and autofluorescence imaging (AFI) 37 , has been evaluated clinically, but is intrinsically low in contrast and based on nonspecific mechanisms
In clinical studies, these methods have not shown clear advantages over conventional white-light endoscopy and random biopsies

Method used

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  • Fibroblast growth factor receptor 2-specific peptide reagents and methods
  • Fibroblast growth factor receptor 2-specific peptide reagents and methods
  • Fibroblast growth factor receptor 2-specific peptide reagents and methods

Examples

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example

[0087] The present invention will be more fully understood by reference to the following examples, which detail exemplary embodiments of the invention.

[0088]The incidence of esophageal adenocarcinoma (EAC) is rapidly increasing, and early detection of Barrett's esophagus (BE) precursor state is challenged by premalignant lesions that are difficult to detect with conventional endoscopic surveillance. Expression of fibroblast growth factor receptor 2 (FGFR2) is an early event in the development of BE to EAC and is a promising imaging target. The peptide SRRPASFRTARE that specifically binds the extracellular domain of FGFR2 was identified using phage display as described in the Examples below. This peptide was labeled with the near-infrared fluorophore Cy5.5, and specific binding to FGFR2 overexpressed in cells was confirmed in vitro. High affinity k d =67.94nM and fast binding k=0.16min -1 (6.2 minutes) was found. In human esophageal samples, peptide agents were found to ...

example 1

[0092] Expression of FGFR2

[0093] Classified by an expert gastrointestinal pathologist (HDA) including squamous (SQ), Barrett's esophagus (BE), low-grade dysplasia (LGD), high-grade dysplasia (HGD) and esophageal adenocarcinoma (EAC ) human esophageal samples were subjected to immunohistochemistry (IHC) to demonstrate representative levels of FGFR2 expression, Figure 6 .

example 2

[0095] FGFR2-specific peptides

[0096] Identification of FGFR2-specific peptides using phage display technology.

[0097] The extracellular domain (ECD) of FGFR2 consists of a signal peptide (SP) and three extracellular immunoglobulin-like domains (D1-D3), Figure 7 a. Peptide selection was performed using the extracellular domain (ECD) of FGFR2c. This area of ​​the target is easy to image. After removing the signal peptide (#10824-H08H-50, Sino Biological), a recombinant FGFR2-ECD (Met1-Glu377) consisting of 367 amino acids was obtained. 1 μg of FGFR2-ECD was subjected to SDS-PAGE using 0.25, 0.5 and 1 μg of BSA as controls to assess quality and quantity. FGFR2-ECD with HPLC purity >97% was used. SDS-PAGE showed an apparent molecular weight of about 65-75 kDa, Figure 7 b. This result is slightly higher than the expected value of 41 kDa due to glycosylation.

[0098] Peptide selection was performed using a phage display library (New England Biolabs, Ph.D.-12) followi...

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Abstract

The present disclosure relates to Fibroblast Growth Factor Receptor 2-specific peptide reagents, methods for detecting epithelial-derived cancer cells such as esophageal, colorectal, gastric, pancreatic or breast carcinoma cells using the peptide reagents, and methods for targeting such cells using the peptide reagents.

Description

[0001] This application claims priority to US Provisional Patent Application Serial No. 62 / 523,446, filed June 22, 2017, which is hereby incorporated by reference in its entirety. [0002] Statement of Government Support [0003] This invention was made with government support under contract U54 CA163059 and U01 CA189291 awarded by the National Institute of Health. The government has certain rights in this invention. [0004] Incorporated by reference into the sequence listing [0005] This application contains the Sequence Listing in computer readable form as a separate part of the disclosure (filename: 52083PCT_SeqList.txt; 1,080 bytes - ASCII text file, created June 20, 2018), which is incorporated by reference in its entirety This article. technical field [0006] The present disclosure relates to fibroblast growth factor receptor 2 (FGFR2) specific peptide reagents; detection of cancers of epithelial origin such as esophageal cancer cells, colorectal cancer cells, g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K45/06C07K7/06
CPCA61K49/0056A61K49/221A61K49/14A61K51/08A61K31/555A61K31/337A61K33/243A61K31/513A61K31/7068A61K31/44A61K31/4745A61K31/196A61K9/5169A61K9/1075A61K47/42A61P35/00A61P35/04C07K7/08C07K1/13A61K49/0032A61K49/0041
Inventor T·D·王J·周
Owner RGT UNIV OF MICHIGAN
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