Anti-COVID-19 Activity of FDA Approved Drugs through RNA G-quadruplex Binding
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
The COVID-19 pandemic caused by SARS-CoV-2 has caused millions of infections and deaths worldwide. Limited treatment options and the threat from emerging variants underline the need for novel and widely accessible therapeutics. G-quadruplexes (G4s) are nucleic acid secondary structures known to affect many cellular processes including viral replication and transcription. We identified heretofore not reported G4s with remarkably low mutation frequency across >5 million SARS-CoV-2 genomes. The G4 structure was targeted using FDA-approved drugs that can bind G4s - Chlorpromazine (CPZ) and Prochlorperazine (PCZ). We found significant inhibition in lung pathology and lung viral load of SARS-CoV-2 challenged hamsters when treated with CPZ, PCZ that was comparable to the widely used antiviral drug Remdesivir. In support, in vitro G4 binding, inhibition of reverse transcription from RNA isolated from COVID-infected humans, and attenuated viral replication and infectivity in Vero cell cultures were clear in case of both CPZ/PCZ. Apart from the wide accessibility of CPZ/PCZ, targeting relatively invariant nucleic acid structures poses an attractive strategy against fast mutating viruses like SARS-CoV-2.
Article activity feed
-
-
SciScore for 10.1101/2022.05.31.493843: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: The same experiment was also done using RNA isolated from nasopharyngeal swabs of COVID-19 infected individuals following approvals by the Institutional Human Ethics Committee (IGIB) Cell culture: All inhibition experiments were carried out on African green monkey kidney cells (Vero CCL-81).
Euthanasia Agents: All the animals, except unchallenged control, were challenged with 105 PFU of SARS-CoV2 administered intranasally using a catheter while under anesthesia by using ketamine (150mg/kg) and xylazine (10mg/kg) intraperitoneal injection inside ABSL3 facility (Chan et al., 2020; Rizvi et al., 2021; Sia et al., 2020).
IACUC: All the experimental protocols involving the handling of virus …SciScore for 10.1101/2022.05.31.493843: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: The same experiment was also done using RNA isolated from nasopharyngeal swabs of COVID-19 infected individuals following approvals by the Institutional Human Ethics Committee (IGIB) Cell culture: All inhibition experiments were carried out on African green monkey kidney cells (Vero CCL-81).
Euthanasia Agents: All the animals, except unchallenged control, were challenged with 105 PFU of SARS-CoV2 administered intranasally using a catheter while under anesthesia by using ketamine (150mg/kg) and xylazine (10mg/kg) intraperitoneal injection inside ABSL3 facility (Chan et al., 2020; Rizvi et al., 2021; Sia et al., 2020).
IACUC: All the experimental protocols involving the handling of virus culture and animal infection were approved by RCGM, institutional biosafety and IAEC animal ethics committee.Sex as a biological variable Animals: 6-8 weeks old male golden Syrian hamsters were procured from CDRI and transported to small animal facility (SAF), THSTI and quarantined for 7 days. Randomization SARS-CoV2 infection in golden Syrian hamster and dosing: Golden Syrian hamsters were randomly allotted to different drug groups (n=4), challenge control (n=2), remdesivir control (n=2) and unchallenged control (n=2) were housed in separate cages. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Briefly Vero cells were seeded in 12-well plate at 90% confluency. Verosuggested: NoneVirus generation for animal experiments: SARS-Related Coronavirus 2, Isolate USA-WA1/2020 virus was grown and titrated in Vero E6 cell line cultured in Dulbecco’s Modified Eagle Medium (DMEM) complete media containing 4.5 g/L D-glucose, 100,000 U/L Penicillin-Streptomycin, 100 mg/L sodium pyruvate, 25mM HEPES and 2% FBS. Vero E6suggested: NoneRecombinant DNA Sentences Resources Reverse transcription was done using reverse primers such that ORF1 pG4-1 or a non G4 forming control region gets reverse transcribed (primer sequences below). pG4-1suggested: NoneThe efficiency of the reverse transcription reactions was measured by quantifying the generated cDNA by qPCR using primers overlapping the ORF1 pG4 or the non G4 forming control region (primer sequences below). pG4suggested: RRID:Addgene_162605)The pre-treatment group viz pCPZ & pPCZ started receiving 8mg/kg and 5mg/kg (respectively) of the drug through intraperitoneal administration each day starting from 3 days prior to the challenge and continued till end point (day 4 post infection). pCPZsuggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
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 did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- No funding statement was detected.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
-