Abstract
Background: As the causative agent of COVID-19, SARS-CoV-2 is a pathogen of immense importance to global public health. Development of innovative direct-acting antiviral agents is sorely needed to address this virus. Peptide-conjugated morpholino oligomers (PPMO) are antisense compounds composed of a phosphorodiamidate morpholino oligomer covalently conjugated to a cell-penetrating peptide. PPMO require no delivery assistance to enter cells and are able to reduce expression of targeted RNA through sequence-specific steric blocking.
Methods: Five PPMO designed against sequences of genomic RNA in the SARS-CoV-2 5′-untranslated region and a negative control PPMO of random sequence were synthesized. Each PPMO was evaluated for its effect on the viability of uninfected cells and its inhibitory effect on the replication of SARS-CoV-2 in Vero-E6 cell cultures. Cell viability was evaluated with an ATP-based method using a 48 h PPMO treatment time. Viral growth was measured with quantitative RT-PCR and TCID50 infectivity assays from experiments where cells received a 5 h PPMO treatment time.
Results: PPMO designed to base-pair with sequence in the 5′ terminal region or the leader transcription regulatory sequence region of SARS-CoV-2 genomic RNA were highly efficacious, reducing viral titres by up to 4-6 log10 in cell cultures at 48-72 h post-infection, in a non-toxic and dose-responsive manner.
Conclusions: The data indicate that PPMO have the ability to potently and specifically suppress SARS-CoV-2 growth and are promising candidates for further preclinical development.
【초록키워드】 COVID-19, Treatment, SARS-CoV-2, peptide, RT-PCR, virus, RNA, Replication, Viral, pathogen, viability, Cell culture, Antiviral agents, experiment, antiviral agent, development, Quantitative, cell viability, genomic RNA, causative agent, Candidates, Post-infection, Quantitative RT-PCR, global public health, growth, sequences, Compound, sequence, inhibitory effect, candidate, titre, random, viral growth, cell cultures, direct-acting, uninfected cells, replication of SARS-CoV-2, VERO-E6, Cell, TCID50, reduce expression, negative control, base-pair, non-toxic, transcription regulatory sequence, oligomer, was measured, composed, evaluated, reducing, suppress, steric, infectivity assay, the SARS-CoV-2, uninfected cell, 【제목키워드】 SARS-CoV-2, inhibition, Culture, Vero Cell, oligomer,