Identification of Anti-Severe Acute Respiratory Syndrome-Related Coronavirus 2 (SARS-CoV-2) Oxysterol Derivatives In Vitro

• Published in: International journal of molecular sciences Vol. 22; no. 6; p. 3163
• Main Authors: Ohashi, Hirofumi, Wang, Feng, Stappenbeck, Frank, Tsuchimoto, Kana, Kobayashi, Chisa, Saso, Wakana, Kataoka, Michiyo, Yamasaki, Masako, Kuramochi, Kouji, Muramatsu, Masamichi, Suzuki, Tadaki, Sureau, Camille, Takeda, Makoto, Wakita, Takaji, Parhami, Farhad, Watashi, Koichi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 19.03.2021
• Subjects: Administration, Oral; Animals; Antiviral Agents - administration & dosage; Antiviral Agents - chemistry; Antiviral Agents - pharmacokinetics; Antiviral Agents - pharmacology; Cell Survival - drug effects; Chlorocebus aethiops; COVID-19 - drug therapy; Mice; Nucleocapsid Proteins - drug effects; Oxysterols - administration & dosage; Oxysterols - chemistry; Oxysterols - pharmacokinetics; Oxysterols - pharmacology; SARS-CoV-2 - drug effects; SARS-CoV-2 - genetics; Vero Cells; Viral Replication Compartments - drug effects; Virus Replication - drug effects; COVID-19; replication; SARS-CoV-2; double membrane vesicle; oxysterols; antiviral; pharmacokinetics; coronavirus
• ISSN: 1422-0067; 1422-0067
• DOI: 10.3390/ijms22063163

The development of effective antiviral drugs targeting the severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) is urgently needed to combat the coronavirus disease 2019 (COVID-19). We have previously studied the use of semi-synthetic derivatives of oxysterols, oxidized derivatives of cholesterol as drug candidates for the inhibition of cancer, fibrosis, and bone regeneration. In this study, we screened a panel of naturally occurring and semi-synthetic oxysterols for anti-SARS-CoV-2 activity using a cell culture infection assay. We show that the natural oxysterols, 7-ketocholesterol, 22(R)-hydroxycholesterol, 24(S)-hydroxycholesterol, and 27-hydroxycholesterol, substantially inhibited SARS-CoV-2 propagation in cultured cells. Among semi-synthetic oxysterols, Oxy210 and Oxy232 displayed more robust anti-SARS-CoV-2 activities, reducing viral replication more than 90% at 10 μM and 99% at 15 μM, respectively. When orally administered in mice, peak plasma concentrations of Oxy210 fell into a therapeutically relevant range (19 μM), based on the dose-dependent curve for antiviral activity in our cell-based assay. Mechanistic studies suggest that Oxy210 reduced replication of SARS-CoV-2 by disrupting the formation of double-membrane vesicles (DMVs); intracellular membrane compartments associated with viral replication. Our study warrants further evaluation of Oxy210 and Oxy232 as a safe and reliable oral medication, which could help protect vulnerable populations with increased risk of developing COVID-19.