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Published on: December 21, 2019
Identification and validation of potent inhibitor of Escherichia coli DHFR from MMV pathogen box
Shingini Sharma1,2, Rashmi Tyagi1, Mitul Srivastava3
1Centre for Drug Design Discovery and Development (C4D), SRM University, Sonepat, Haryana, India.
Abstract:
The present study is conducted to find the solution of rising antimicrobial resistance (AMR) in Escherichia coli which is a pathogen responsible for fatal systemic infections in human and animals. The enzyme dihydrofolate reductase (DHFR) is found in all organisms. In this study DHFR of E. coli (ec-DHFR) and human DHFR (h-DHFR) is targeted by novel chemical entities (NCE) from the Pathogen box of Medicines for Malaria Venture, Switzerland (MMV) using molecular modelling. The in-silico studies were further validated by in-vitro assays. The virtual screening of 400 MMV compounds was conducted using PyRx standard tool followed by manual docking of selected compounds by Autodock vina and Ligplot program. The in-silico studies showed good binding energy and strong hydrogen bond in docking of MMV675968 with ec-DHFR and no hydrogen bond with h-DHFR. This was further validated by the Molecular dynamic studies that revealed high binding free energy in ec-DHFR and in-vitro assays that produced good synergy in combination study of MMV675968 with last line (meropenem) and last resort (colistin) antibiotics. The extensive MD simulation and energetic analysis thus concludes that MMV675968 targets ec-DHFR. The combination studies were conducted with MMV675968 and FDA approved drugs against a panel of multidrug resistant Escherichia coli isolates. The synergistic results obtained in combination studies concluded that in-vitro data is consistent with in-silico data and that MMV675968 is a potential lead for future process of antimicrobial drug development against the multidrug resistance E. coli.Communicated by Ramaswamy H. Sarma.
Insights
Researchers identified MMV675968 as a potential antimicrobial drug candidate targeting dihydrofolate reductase in antimicrobial-resistant Escherichia coli. In-vitro studies confirmed its synergistic effect with existing antibiotics, showing promise for developing new treatments against multidrug-resistant infections.
Area of Science:
- Medicinal Chemistry
- Computational Biology
- Microbiology
Background:
- Antimicrobial resistance (AMR) in Escherichia coli poses a significant threat to human and animal health.
- Dihydrofolate reductase (DHFR) is a crucial enzyme in all organisms and a viable drug target.
Purpose of the Study:
- To identify novel chemical entities (NCEs) targeting ec-DHFR for combating AMR in E. coli.
- To validate in-silico findings with in-vitro experimental data.
Main Methods:
- Virtual screening of 400 compounds from the Medicines for Malaria Venture (MMV) Pathogen box using PyRx.
- Molecular docking using Autodock vina and Ligplot.
- Molecular dynamic (MD) simulations and in-vitro assays.
Main Results:
- MMV675968 showed strong binding affinity and hydrogen bonding with ec-DHFR but not h-DHFR.
- MD studies confirmed high binding free energy with ec-DHFR.
- In-vitro assays demonstrated synergistic effects of MMV675968 in combination with meropenem and colistin against multidrug-resistant E. coli.
Conclusions:
- MMV675968 effectively targets ec-DHFR.
- In-silico and in-vitro data are consistent, supporting MMV675968 as a promising lead compound for developing new antimicrobial drugs against multidrug-resistant E. coli.
