Synthesis and In-Vitro Inhibition Assay of Cation-Pi Peptide Targeting Main Protease of SARS-CoV-2
Disciplines
Biochemistry | Medicinal-Pharmaceutical Chemistry
Abstract (300 words maximum)
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro) has been identified as an essential enzyme for viral replication and transcription, making it a key target for therapeutic intervention in COVID-19 treatment. In this study, three peptides: SLFWQWKSKFLGR, SLFWQWHSKFLGR, and SLFWQWRSKFLGR were synthesized to inhibit Mpro activity through cation- interactions. The attractive force between the positive charge of the basic amino acids, the side chain of lysine (K), histidine (H), and arginine (R) with the electron-rich -system of an aromatic ring of phenylalanine (F). This cation- interaction plays a key role in stabilizing molecular structures, particularly in biological systems, and can be critical for enzyme inhibition. The peptides were engineered based on their ability to form stable non-covalent interactions with Cys145 and His41 residues present in the active site of Mpro, with the aim of blocking the proteolytic activity necessary for viral polyprotein processing. The three peptides were synthesized using solid-phase peptide synthesis (SPPS) and characterized by mass spectrometry, which confirmed its expected molecular weight and purity. Once synthesized and validated, all three peptides were evaluated in biological assays to determine their inhibitory potential against SARS-CoV-2 Mpro. Each assay was repeated three times to ensure reproducibility, consistency, and statistical significance. Additionally, a fluorescence resonance anergy transfer (FRET) assay was conducted for all three peptides to further assess their binding interactions and efficacy against Mpro. The FRET assay evaluates the binding interactions of the peptides with Mpro by monitoring energy transfer between donor and acceptor fluorophores. Through this research, the potential of cation- interaction-based peptides as inhibitors of viral proteases was explored, with a focus on targeting Mpro as a therapeutic strategy. The successful inhibition of Mpro could provide a promising approach for developing antiviral therapies against SARS-CoV-2.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad Halim
Synthesis and In-Vitro Inhibition Assay of Cation-Pi Peptide Targeting Main Protease of SARS-CoV-2
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro) has been identified as an essential enzyme for viral replication and transcription, making it a key target for therapeutic intervention in COVID-19 treatment. In this study, three peptides: SLFWQWKSKFLGR, SLFWQWHSKFLGR, and SLFWQWRSKFLGR were synthesized to inhibit Mpro activity through cation- interactions. The attractive force between the positive charge of the basic amino acids, the side chain of lysine (K), histidine (H), and arginine (R) with the electron-rich -system of an aromatic ring of phenylalanine (F). This cation- interaction plays a key role in stabilizing molecular structures, particularly in biological systems, and can be critical for enzyme inhibition. The peptides were engineered based on their ability to form stable non-covalent interactions with Cys145 and His41 residues present in the active site of Mpro, with the aim of blocking the proteolytic activity necessary for viral polyprotein processing. The three peptides were synthesized using solid-phase peptide synthesis (SPPS) and characterized by mass spectrometry, which confirmed its expected molecular weight and purity. Once synthesized and validated, all three peptides were evaluated in biological assays to determine their inhibitory potential against SARS-CoV-2 Mpro. Each assay was repeated three times to ensure reproducibility, consistency, and statistical significance. Additionally, a fluorescence resonance anergy transfer (FRET) assay was conducted for all three peptides to further assess their binding interactions and efficacy against Mpro. The FRET assay evaluates the binding interactions of the peptides with Mpro by monitoring energy transfer between donor and acceptor fluorophores. Through this research, the potential of cation- interaction-based peptides as inhibitors of viral proteases was explored, with a focus on targeting Mpro as a therapeutic strategy. The successful inhibition of Mpro could provide a promising approach for developing antiviral therapies against SARS-CoV-2.