Synthesis, Characterization, and In-Vitro Analysis of Antiviral Peptides Targeting the Spike Protein of Sars-CoV-2 and Angiotensin Converting Enzyme 2
Disciplines
Analytical Chemistry | Biochemistry | Computational Chemistry | Organic Chemistry
Abstract (300 words maximum)
The development of therapies to treat Covid-19, a potentially fatal disease that has ended the lives of over 6 million people as of October 2022, is important as ever due to the disease’s prevalence in almost every nation in the world despite there being widely available vaccines. Sars-CoV-2, the causative agent of Covid-19, possesses a spike protein that serves as an apt vector for inhibition due to its key role in cellular penetration. The spike protein is found on the viral envelope and aids in membrane fusion by targeting angiotensin converting enzyme 2, an enzyme found on the surface of many human cells. The spike protein’s active site has a large surface area devoted to protein-protein interaction. This large surface area is not conducive to inhibition by small molecules and thus necessitates the development of macromolecular therapeutics. Peptides are not hindered by large active sites and thus are advantageous in this scenario. Furthermore, strong specificity and low production costs make peptides valuable therapeutic assets. Previous studies have identified strong candidate peptides using in-silico analysis. Several of these peptides were chosen to be synthesized by utilizing automated Fmoc solid-phase synthesis. Peptide products were characterized using mass spectroscopy, ensuring experimental m/z values agreed with theoretical values. A competitive ELISA was utilized to assess the effectiveness of peptides in disrupting the spike-ACE2 complex. AH33, a biomimetic peptide that emulates the ACE2 active site showed the greatest promise as an inhibitor for the spike-ACE2 complex. It is identified by an intense peak at 1314.28 m/z which is assigned to the [M+3H]3+ ion. When tested for activity AH33 showed 40% inhibition at 100µM and 30% inhibition at 20µM. This suggests further development of biomimetic peptides could yield fruitful inhibitors for the spike-ACE2 complex with possible modifications such as stapling and cyclization.
Academic department under which the project should be listed
Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad Halim
Synthesis, Characterization, and In-Vitro Analysis of Antiviral Peptides Targeting the Spike Protein of Sars-CoV-2 and Angiotensin Converting Enzyme 2
The development of therapies to treat Covid-19, a potentially fatal disease that has ended the lives of over 6 million people as of October 2022, is important as ever due to the disease’s prevalence in almost every nation in the world despite there being widely available vaccines. Sars-CoV-2, the causative agent of Covid-19, possesses a spike protein that serves as an apt vector for inhibition due to its key role in cellular penetration. The spike protein is found on the viral envelope and aids in membrane fusion by targeting angiotensin converting enzyme 2, an enzyme found on the surface of many human cells. The spike protein’s active site has a large surface area devoted to protein-protein interaction. This large surface area is not conducive to inhibition by small molecules and thus necessitates the development of macromolecular therapeutics. Peptides are not hindered by large active sites and thus are advantageous in this scenario. Furthermore, strong specificity and low production costs make peptides valuable therapeutic assets. Previous studies have identified strong candidate peptides using in-silico analysis. Several of these peptides were chosen to be synthesized by utilizing automated Fmoc solid-phase synthesis. Peptide products were characterized using mass spectroscopy, ensuring experimental m/z values agreed with theoretical values. A competitive ELISA was utilized to assess the effectiveness of peptides in disrupting the spike-ACE2 complex. AH33, a biomimetic peptide that emulates the ACE2 active site showed the greatest promise as an inhibitor for the spike-ACE2 complex. It is identified by an intense peak at 1314.28 m/z which is assigned to the [M+3H]3+ ion. When tested for activity AH33 showed 40% inhibition at 100µM and 30% inhibition at 20µM. This suggests further development of biomimetic peptides could yield fruitful inhibitors for the spike-ACE2 complex with possible modifications such as stapling and cyclization.