Investigating the Inhibitory Potential of the Frog-Skin Derived Peptide DRAMP01292 Against the SARS-CoV-2 Main Protease
Disciplines
Medicinal-Pharmaceutical Chemistry | Organic Chemistry
Abstract (300 words maximum)
The COVID-19 pandemic was caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), resulting in millions of deaths around the world and still imposes a global health burden. Current strategies for targeting SARS-CoV-2 are focused on vaccines and small-molecule antivirals. However, these methods can have limited efficiency due to drug resistance and off-target toxicity – highlighting a need for other approaches. Peptide therapeutics are an excellent alternative as they are highly selective, can be chemically modified, and interact with a larger surface area. This study investigated the frog-skin derived peptide DRAMP01292 for its use in inhibiting Mpro activity. DRAMP01292 was synthesized using Fmoc-based solid-phase peptide synthesis (SPPS). Deprotection and cleavage from the resin were performed with the use of TFA. The crude peptide was then purified using reversed-phase high performance liquid chromatography (RP-HPLC). Liquid Chromatography–Mass Spectroscopy (LC-MS) analysis confirmed the expected molecular weight (2113.533 Da), with the peptide eluting at 1.30 min on a C18 column. The mass spectrum displayed peaks at m/z 1057.08 ([M+2H] ²⁺), 705.05 ([M+3H] ³⁺), and 529.04 ([M+4H] ⁴⁺). To assess the inhibitory efficiency of DRAMP01292 against SARS-CoV-2 Mpro, a Fluorescence Resonance Energy Transfer (FRET) cleavage-based assay was performed. The assay measures fluorescence upon substrate cleavage, providing information on Mpro’s enzymatic activity in the presence or absence of the inhibitor. The initial FRET reading resulted in an IC₅₀ of 21.95 µM, meaning the concentration required to inhibit enzyme activity by 50%. Although this value represents moderate inhibition compared to small molecules, it shows that DRAMP01292 can inhibit Mpro, and further testing can result in increased inhibition efficiency. Overall, this study highlights the potential of peptide therapeutics to strengthen and diversify existing antiviral strategies against SARS-CoV-2.
Use of AI Disclaimer
no
Academic department under which the project should be listed
CSM – Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad Halim
Investigating the Inhibitory Potential of the Frog-Skin Derived Peptide DRAMP01292 Against the SARS-CoV-2 Main Protease
The COVID-19 pandemic was caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), resulting in millions of deaths around the world and still imposes a global health burden. Current strategies for targeting SARS-CoV-2 are focused on vaccines and small-molecule antivirals. However, these methods can have limited efficiency due to drug resistance and off-target toxicity – highlighting a need for other approaches. Peptide therapeutics are an excellent alternative as they are highly selective, can be chemically modified, and interact with a larger surface area. This study investigated the frog-skin derived peptide DRAMP01292 for its use in inhibiting Mpro activity. DRAMP01292 was synthesized using Fmoc-based solid-phase peptide synthesis (SPPS). Deprotection and cleavage from the resin were performed with the use of TFA. The crude peptide was then purified using reversed-phase high performance liquid chromatography (RP-HPLC). Liquid Chromatography–Mass Spectroscopy (LC-MS) analysis confirmed the expected molecular weight (2113.533 Da), with the peptide eluting at 1.30 min on a C18 column. The mass spectrum displayed peaks at m/z 1057.08 ([M+2H] ²⁺), 705.05 ([M+3H] ³⁺), and 529.04 ([M+4H] ⁴⁺). To assess the inhibitory efficiency of DRAMP01292 against SARS-CoV-2 Mpro, a Fluorescence Resonance Energy Transfer (FRET) cleavage-based assay was performed. The assay measures fluorescence upon substrate cleavage, providing information on Mpro’s enzymatic activity in the presence or absence of the inhibitor. The initial FRET reading resulted in an IC₅₀ of 21.95 µM, meaning the concentration required to inhibit enzyme activity by 50%. Although this value represents moderate inhibition compared to small molecules, it shows that DRAMP01292 can inhibit Mpro, and further testing can result in increased inhibition efficiency. Overall, this study highlights the potential of peptide therapeutics to strengthen and diversify existing antiviral strategies against SARS-CoV-2.