ACIS, A Novel KepTide™, Binds to ACE-2 Receptor and Inhibits the Infection of SARS-CoV2 Virus in vitro in Primate Kidney Cells: Therapeutic Implications for COVID-19
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
Coronavirus disease 2019 (COVID-19) is a severe acute respiratory syndrome (SARS) caused by a virus known as SARS-Coronavirus 2 (SARS-CoV2). Without a targeted-medicine, this disease has been causing a massive humanitarian crisis not only in terms of mortality, but also imposing a lasting damage to social life and economic progress of humankind. Therefore, an immediate therapeutic strategy needs to be intervened to mitigate this global crisis. Here, we report a novel KepTide™ (Knock-End Peptide) therapy that nullifies SARS-CoV2 infection. SARS-CoV2 employs its surface glycoprotein “spike” (S-glycoprotein) to interact with angiotensin converting enzyme-2 (ACE-2) receptor for its infection in host cells. Based on our in-silico -based homology modeling study validated with a recent X-ray crystallographic structure (PDB ID:6M0J), we have identified that a conserved motif of S-glycoprotein that intimately engages multiple hydrogen-bond (H-bond) interactions with ACE-2 enzyme. Accordingly, we designed a peptide, termed as ACIS ( AC E-2 I nhibitory motif of S pike), that displayed significant affinity towards ACE-2 enzyme as confirmed by biochemical assays such as BLItz and fluorescence polarization assays. Interestingly, more than one biochemical modifications were adopted in ACIS in order to enhance the inhibitory action of ACIS and hence called as KEpTide™. Consequently, a monolayer invasion assay, plaque assay and dual immunofluorescence analysis further revealed that KEpTide™ efficiently mitigated the infection of SARS-CoV2 in vitro in VERO E6 cells. Finally, evaluating the relative abundance of ACIS in lungs and the potential side-effects in vivo in mice, our current study discovers a novel KepTide™ therapy that is safe, stable, and robust to attenuate the infection of SARS-CoV2 virus if administered intranasally.
Article activity feed
-
SciScore for 10.1101/2020.10.13.337584: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
NIH rigor criteria are not applicable to paper type.Table 2: Resources
Antibodies Sentences Resources S-glycoprotein antibody (Anti-SARS Antibody, S Specific, clone 154C; Cat # MAB8783) was purchased from Millipore Sigma Anti-SARSsuggested: NoneDonkey anti-mouse Alexa 647-conjugated affinity pure secondary antibody (Cat # 715-605-150) was purchased from Jackson ImmunoResearch Laboratories Inc. Spike RBD (SARS-CoV-2): anti-mousesuggested: (Jackson ImmunoResearch Labs Cat# 715-605-150, RRID:AB_2340862)Experimental Models: Cell Lines Sentences Resources VERO E6 cells were pre-incubated with DMSO, 25 μM of KEpTide 1 and 2 for 30 mins and then inoculated with 500 μL of each SARS-CoV2 dilution and incubated at … SciScore for 10.1101/2020.10.13.337584: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
NIH rigor criteria are not applicable to paper type.Table 2: Resources
Antibodies Sentences Resources S-glycoprotein antibody (Anti-SARS Antibody, S Specific, clone 154C; Cat # MAB8783) was purchased from Millipore Sigma Anti-SARSsuggested: NoneDonkey anti-mouse Alexa 647-conjugated affinity pure secondary antibody (Cat # 715-605-150) was purchased from Jackson ImmunoResearch Laboratories Inc. Spike RBD (SARS-CoV-2): anti-mousesuggested: (Jackson ImmunoResearch Labs Cat# 715-605-150, RRID:AB_2340862)Experimental Models: Cell Lines Sentences Resources VERO E6 cells were pre-incubated with DMSO, 25 μM of KEpTide 1 and 2 for 30 mins and then inoculated with 500 μL of each SARS-CoV2 dilution and incubated at 37 °C for 1 h with rocking every 15 min. VERO E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Bioavailability assay of KEpTide: Eight to ten weeks-old BALB/C mouse were administered with KEpTide 1 at a dose of 0.1 mg/Kg body weight intranasally. BALB/Csuggested: RRID:IMSR_ORNL:BALB/cRl)Software and Algorithms Sentences Resources The sequence of S-glycoprotein was derived in FASTA format from the sequence available in PubMed with accession number QIC53213.1 and GI number 1811294675. PubMedsuggested: (PubMed, RRID:SCR_004846)The most stable docked-structure had been resolved based on electrostatic (Eele), desolvation (Edesolv) and Van der Waals (ΔGVDW) energies and finally displayed with Chimera software. Chimerasuggested: (Chimera, RRID:SCR_002959)All curve fitting and data analysis were performed using Igor Pro (Wave Metrics, Portland, OR). Igor Prosuggested: (IGOR Pro, RRID:SCR_000325)Curve-fitting was done in GraphPad Prism 8 software using log[inhibitor] versus response, variable slope liner regression curve analysis module and resultant IC50 value was calculated. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)The endpoint absorbance wavelength was 500 nm, and the final absorbance was recorded in Gen5 software version 2.05. Gen5suggested: (Gen5, RRID:SCR_017317)Results from OddPub: Thank you for sharing your data.
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 29 and 30. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
-