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Published on: November 11, 2016
Identification of small molecule inhibitors against MMP-14 via High-Throughput screening
Hyun Lee1, Isoo Youn2, Robel Demissie1
1Department of Pharmaceutical Sciences, College of Pharmacy, University of Illinois at Chicago, Chicago, IL 60607, USA; Biophysics Core at Research Resource Center, University of Illinois at Chicago, Chicago, IL 60607, USA.
Abstract:
Matrix metalloproteinases (MMPs) are involved in various cellular events in physiology and pathophysiology through endopeptidases activity. The expression levels and activities of most MMPs remain minimal in the normal conditions, whereas some MMPs are significantly activated in pathological conditions such as cancer and neovascularization. Hence, MMPs are considered as both diagnostic markers and potential targets for therapeutic agents. Twenty-three known human MMPs share a similar active site structure with a zinc-binding motif, resulting in lack of specificity. Therefore, the enhancement of target specificity is a primary goal for the development of specific MMP inhibitors. MMP-14 regulates VEGFA/VEGFR2-system through cleavage of the non-functional VEGFR1 in vascular angiogenesis. In this study, we developed a fluorescence-based enzymatic assay using a specific MMP-14 substrate generated from VEGFR1 cleavage site. This well optimized assay was used as a primary screen method to identify MMP-14 specific inhibitors from 1,200 Prestwick FDA-approved drug library. Of ten initial hits, two compounds showed IC50 values below 30 µM, which were further validated by direct binding analysis using surface plasmon resonance (SPR). Clioquinol and chloroxine, both of which contain a quinoline structure, were identified as MMP-14 inhibitors. Five analogs were tested, four of which were found to be completely devoid of inhibitory activity. Clioquinol exhibited selectivity towards MMP-14, as it showed no inhibitory activity towards four other MMPs.
Insights
Researchers identified specific inhibitors for Matrix Metalloproteinase-14 (MMP-14), a key enzyme in angiogenesis. Clioquinol and chloroxine were found to inhibit MMP-14, with clioquinol showing selectivity.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Matrix metalloproteinases (MMPs) are crucial enzymes in physiological and pathological processes, including cancer and neovascularization.
- The lack of specificity among MMPs hinders their therapeutic potential, necessitating the development of targeted inhibitors.
- MMP-14 plays a significant role in vascular angiogenesis by regulating the VEGFA/VEGFR2 system.
Purpose of the Study:
- To develop a specific assay for screening MMP-14 inhibitors.
- To identify specific MMP-14 inhibitors from a library of FDA-approved drugs.
- To evaluate the selectivity of identified inhibitors against other MMPs.
Main Methods:
- Development of a fluorescence-based enzymatic assay using a MMP-14 specific substrate derived from VEGFR1.
- Screening of the Prestwick FDA-approved drug library (1,200 compounds).
- Validation of initial hits using surface plasmon resonance (SPR) and testing of analogs.
Main Results:
- A specific and optimized fluorescence-based assay for MMP-14 was established.
- Ten initial hits were identified, with two compounds (clioquinol and chloroxine) showing IC50 values below 30 µM.
- Clioquinol demonstrated selectivity for MMP-14, with no observed inhibition against four other tested MMPs.
Conclusions:
- Clioquinol and chloroxine are identified as MMP-14 inhibitors.
- The developed assay is effective for primary screening of MMP-14 inhibitors.
- Clioquinol represents a promising candidate for further investigation as a selective MMP-14 inhibitor.
