Potential for Alpha-Conotoxins Peptides as Inhibitors of Main Protease in SARS-CoV-2
Disciplines
Biochemistry
Abstract (300 words maximum)
Main protease (Mpro) is an enzyme crucial for replication in SARS-CoV-2, so designing and developing inhibitors targeting this enzyme are of great medical interest for treatment of COVID-19. Peptide therapies have an advantage over other treatment options because they have little to no side effects and can therefore be administered to a wider range of patients. Peptides derived from alpha-conotoxins, a type of venom present in cone snails, have shown potential in the past as treatments for various diseases including neurological, bacterial and viral diseases. However, no studies are conducted using these peptides targeting Mpro of SAS-CoV-2. The aims of these studies are to chemically synthesize alpha-conotoxins peptides and test their inhibition efficiency against the Mpro of SAS-CoV-2. These peptides were synthesized using the standard solid phase peptide synthesis (SPPS) protocol employing a Liberty Blue Microwave Peptide Synthesizer. After synthesizing the peptide on the resin, the peptide-resin was dried using a vacuum filter and cleaved using a cleavage cocktail consisting of 5% phenol, 5% water, 5% thioanisole, 2.5% 1,2-ethanediol, and 82.5% TFA. Excess TFA was evaporated off using N2 gas. The peptide solution was then mixed with ice-cold ether and centrifuged at 7000 rpm for 10 minutes at 4°C. The ether was decanted off and the remaining peptide was dissolved in acetic acid and water. This solution was then frozen at -80°C and lyophilized overnight. The mass of the peptides was confirmed by electrospray ionization coupled with mass spectrometry. The linear peptide showed two strong peaks at m/z 677.33 and 1315.41 corresponded to [M+2H]2+ and [M+H]+ ions, respectively which matched theoretical values. Cyclic peptide was synthesized by adding 10% DMSO and stirred for 24-48 hours. Cyclization was confirmed by mass spectrometry which showed the removal of four hydrogen from four cysteine residues and forming disulfide bonds between Cys2-Cys8 and Cys3-Cys13.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad A. Halim
Potential for Alpha-Conotoxins Peptides as Inhibitors of Main Protease in SARS-CoV-2
Main protease (Mpro) is an enzyme crucial for replication in SARS-CoV-2, so designing and developing inhibitors targeting this enzyme are of great medical interest for treatment of COVID-19. Peptide therapies have an advantage over other treatment options because they have little to no side effects and can therefore be administered to a wider range of patients. Peptides derived from alpha-conotoxins, a type of venom present in cone snails, have shown potential in the past as treatments for various diseases including neurological, bacterial and viral diseases. However, no studies are conducted using these peptides targeting Mpro of SAS-CoV-2. The aims of these studies are to chemically synthesize alpha-conotoxins peptides and test their inhibition efficiency against the Mpro of SAS-CoV-2. These peptides were synthesized using the standard solid phase peptide synthesis (SPPS) protocol employing a Liberty Blue Microwave Peptide Synthesizer. After synthesizing the peptide on the resin, the peptide-resin was dried using a vacuum filter and cleaved using a cleavage cocktail consisting of 5% phenol, 5% water, 5% thioanisole, 2.5% 1,2-ethanediol, and 82.5% TFA. Excess TFA was evaporated off using N2 gas. The peptide solution was then mixed with ice-cold ether and centrifuged at 7000 rpm for 10 minutes at 4°C. The ether was decanted off and the remaining peptide was dissolved in acetic acid and water. This solution was then frozen at -80°C and lyophilized overnight. The mass of the peptides was confirmed by electrospray ionization coupled with mass spectrometry. The linear peptide showed two strong peaks at m/z 677.33 and 1315.41 corresponded to [M+2H]2+ and [M+H]+ ions, respectively which matched theoretical values. Cyclic peptide was synthesized by adding 10% DMSO and stirred for 24-48 hours. Cyclization was confirmed by mass spectrometry which showed the removal of four hydrogen from four cysteine residues and forming disulfide bonds between Cys2-Cys8 and Cys3-Cys13.