Evaluating Disulfiram's Effects on SARS-CoV-2 in Humanized Mice

To evaluate the effects of Disulfiram in vivo, we infected the K18-hACE2 humanized mice (B6.Cg-Tg(K18-ACE2)2Prlmn/J) (McCray et al. [25 (link)] Oladunni et al. [26 (link)], Bao et al. [27 (link)]). K18-hACE2 mice were obtained from The Jackson Laboratory and were bred in the Centro de Criação de Animais Especiais (Ribeirão Preto Medical School/University of São Paulo). For the experimental infection, animals were transferred to the BSL3 facility. Eight-week-old male K18-hACE2 mice were infected with 2 × 10⁴ PFU of SARS-CoV-2 (in 40 µL) by the intranasal route as previously described [26 (link)]. Twenty-four hours after the virus inoculation and once daily until day 5 post-infection (dpi), animals were treated with Disulfiram (50 mg/kg, i.p.) (n = 7) or vehicle (n = 7). Naive mice remained uninfected and untreated. On the 5 dpi, 6 h after the injection of Disulfiram or vehicle, animals were humanely euthanized for samples collection. All the experimental procedures were performed in accordance with the guide for the use of laboratory animals of the University of Sao Paulo and approved by the institutional ethics committee (066/2020).