Designing Small Peptide Analogues to inhibit the Spike Protein of SARS-CoV-2
Disciplines
Biochemistry | Medicinal-Pharmaceutical Chemistry
Abstract (300 words maximum)
Since 2020, SARS-CoV-2 has affected millions of people. Many pharmaceutical companies have implemented vaccines and booster shots to decrease infection rate and increase the chance of immunity, but recent studies consider the use of antiviral peptides as well. Antiviral peptides can inhibit the spike receptor-binding domain of SARS-CoV-2 and can ease symptoms. The aim of this project was to synthesize the alpha-helix peptide analogues of human ACE2 and determine its efficacy against COVID-19. Various research showed that alpha-helix (AH) of ACE2 showed good inhibition efficiency, however, this peptide has a long sequence of 33 amino acids which is difficult to synthesis by solid phase peptide synthesis. Herein we have designed three shorter analogues based on the C-terminal (CT), Middle (MT), and N-terminal (NT) of the alpha-helix peptide. A solid-phase synthesis protocols was employed to synthesize these peptides using CEM Liberty Blue Peptide Synthesizer. A high-swelling rink amide resin with loading capacity of 0.6 mmol/g and 100-200 mesh size was used for the synthesis. After synthesizing the peptide, the peptide-resin complex was cleaved using 95%TFA. The peptide solution was precipitated with cold ether, subsequently the peptide was dissolved with 10% acetic acid, and lyophilized overnight. The peptides were then characterized by liquid chromatography (LC) and mass spectrometry (MS) experiment. The LC experiment was performed using Agilent 1290 UPLC system using a C8 column. For CT peptides, three peaks were found at m/z 1937.75 Da, 969.50 Da, and 646.75 Da which corresponded to [M+H]+, [M+2H]+, and [M+3H]3+ charge states. The charge states of this peptide exactly matched with the theorical masses. Similar results are also observed for MT and NT peptides. Additionally, biological assays were performed to further show each peptides’ effectiveness. At 100 uM, CT peptides showed 22.32% of inhibition efficiency while MT peptides demonstrated 31.04% of inhibition.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad A. Halim
Designing Small Peptide Analogues to inhibit the Spike Protein of SARS-CoV-2
Since 2020, SARS-CoV-2 has affected millions of people. Many pharmaceutical companies have implemented vaccines and booster shots to decrease infection rate and increase the chance of immunity, but recent studies consider the use of antiviral peptides as well. Antiviral peptides can inhibit the spike receptor-binding domain of SARS-CoV-2 and can ease symptoms. The aim of this project was to synthesize the alpha-helix peptide analogues of human ACE2 and determine its efficacy against COVID-19. Various research showed that alpha-helix (AH) of ACE2 showed good inhibition efficiency, however, this peptide has a long sequence of 33 amino acids which is difficult to synthesis by solid phase peptide synthesis. Herein we have designed three shorter analogues based on the C-terminal (CT), Middle (MT), and N-terminal (NT) of the alpha-helix peptide. A solid-phase synthesis protocols was employed to synthesize these peptides using CEM Liberty Blue Peptide Synthesizer. A high-swelling rink amide resin with loading capacity of 0.6 mmol/g and 100-200 mesh size was used for the synthesis. After synthesizing the peptide, the peptide-resin complex was cleaved using 95%TFA. The peptide solution was precipitated with cold ether, subsequently the peptide was dissolved with 10% acetic acid, and lyophilized overnight. The peptides were then characterized by liquid chromatography (LC) and mass spectrometry (MS) experiment. The LC experiment was performed using Agilent 1290 UPLC system using a C8 column. For CT peptides, three peaks were found at m/z 1937.75 Da, 969.50 Da, and 646.75 Da which corresponded to [M+H]+, [M+2H]+, and [M+3H]3+ charge states. The charge states of this peptide exactly matched with the theorical masses. Similar results are also observed for MT and NT peptides. Additionally, biological assays were performed to further show each peptides’ effectiveness. At 100 uM, CT peptides showed 22.32% of inhibition efficiency while MT peptides demonstrated 31.04% of inhibition.