Abstract
Background. First vaccines for prevention of Coronavirus disease 2019
(COVID-19) are becoming available but there is a huge and unmet need for
specific forms of treatment. In this study we aimed to evaluate the
potent anti-SARS-CoV-2 effect of siRNA both in vitro and in vivo.
Methods. To identify most effective molecule out of a panel of 15 in
silico designed siRNAs, an in vitro screening system based on vectors
expressing SARS-CoV-2 genes fused with the firefly luciferase reporter
gene and SARS-CoV-2-infected cells was used. The most potent siRNA,
siR-7, was modified by Locked nucleic acids (LNAs) to obtain siR-7-EM
with increased stability and was formulated with the peptide dendrimer
KK-46 for enhancing cellular uptake to allow topical application by
inhalation of the final formulation - siR-7-EM/KK-46. Using the Syrian
Hamster model for SARS-CoV-2 infection the antiviral capacity of
siR-7-EM/KK-46 complex was evaluated. Results. We identified the siRNA,
siR-7, targeting SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) as the
most efficient siRNA inhibiting viral replication in vitro. Moreover, we
have shown that LNA-modification and complexation with the designed
peptide dendrimer enhanced the antiviral capacity of siR-7 in vitro. We
demonstrated significant reduction of virus titer and total lung
inflammation in the animals exposed by inhalation of siR-7-EM/KK-46 in
vivo. Conclusions. Thus, we developed a therapeutic strategy for
COVID-19 based on inhalation of a modified siRNA-peptide dendrimer
formulation.