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Updated: Jun 9, 2025
Screening and Identification of Small Peptides Targeting Fibroblast Growth Factor Receptor2 using a Phage Display Peptide Library
Published on: September 30, 2019
Sensitive quantification of fibroblast activation protein and high-throughput screening for inhibition by
Kateřina Čermáková1, Adéla Šimková2, Filip Wichterle3
1Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo náměstí 2, 166 10 Prague 6, Czech Republic; First Faculty of Medicine, Charles University, Kateřinská 32, 121 08 Prague 2, Czech Republic.
Abstract:
Fibroblast activation protein (FAP) has been extensively studied as a cancer biomarker for decades. Recently, small-molecule FAP inhibitors have been widely adopted as a targeting moiety of experimental theranostic radiotracers. Here we present a fast qPCR-based analytical method allowing FAP inhibition screening in a high-throughput regime. To identify clinically relevant compounds that might interfere with FAP-targeted approaches, we focused on a library of FDA-approved drugs. Using the DNA-linked Inhibitor Antibody Assay (DIANA), we tested a library of 2667 compounds within just a few hours and identified numerous FDA-approved drugs as novel FAP inhibitors. Among these, prodrugs of cephalosporin antibiotics and reverse transcriptase inhibitors, along with one elastase inhibitor, were the most potent FAP inhibitors in our dataset. In addition, by employing FAP DIANA in the quantification mode, we were able to determine FAP concentrations in human plasma samples. Together, our work expands the repertoire of FAP inhibitors, analyzes the potential interference of co-administered drugs with FAP-targeting strategies, and presents a sensitive and low-consumption ELISA alternative for FAP quantification with a detection limit of 50 pg/ml.
Insights
This study introduces a rapid assay to screen FDA-approved drugs for inhibiting fibroblast activation protein (FAP). Several drugs, including antibiotics and reverse transcriptase inhibitors, were identified as novel FAP inhibitors, aiding theranostic radiotracer development.
Area of Science:
- Biochemistry
- Pharmacology
- Oncology
Background:
- Fibroblast activation protein (FAP) is a key cancer biomarker.
- FAP inhibitors are crucial for developing targeted theranostic radiotracers.
- Identifying drug interactions with FAP is vital for clinical applications.
Purpose of the Study:
- To develop a high-throughput method for screening FAP inhibitors.
- To identify FDA-approved drugs that inhibit FAP.
- To assess potential interference of existing drugs with FAP-targeted therapies.
Main Methods:
- Developed a fast, qPCR-based DNA-linked Inhibitor Antibody Assay (DIANA) for FAP inhibition screening.
- Screened a library of 2667 FDA-approved drugs.
- Quantified FAP concentrations in human plasma using FAP DIANA.
Main Results:
- Identified numerous FDA-approved drugs as novel FAP inhibitors.
- Prodrugs of cephalosporin antibiotics, reverse transcriptase inhibitors, and an elastase inhibitor showed potent FAP inhibition.
- Achieved a detection limit of 50 pg/ml for FAP quantification, offering an alternative to ELISA.
Conclusions:
- Expands the known repertoire of FAP inhibitors.
- Provides insights into potential drug-drug interactions affecting FAP-targeted theranostics.
- Offers a sensitive, low-consumption method for FAP quantification in biological samples.
