Inhibition of Brazilian ZIKV strain replication in primary human placental chorionic cells and cervical cells treated with nitazoxanide

Abstract

Zika virus (ZIKV) infection during pregnancy is associated with a congenital syndrome. Although the virus can be detected in human placental tissue and sexual transmission has been verified, it is not clear how the virus reaches the fetus. Despite the emerging severity caused by ZIKV infection, no specific prophylactic and/or therapeutic treatment is available. The aim of the present study was to evaluate the effectiveness antiviral of nitazoxanide (NTZ) in two important congenital transmission targets: (i) a primary culture of human placental chorionic cells, and (ii) human cervical epithelial cells (C33-A) infected with Brazilian ZIKV strain. Initially, NTZ activity was screened in ZIKV infected Vero cells under different treatment regimens with non-toxic drug concentrations for 48 h. Antiviral effect was found only when the treatment was carried out after the viral inoculum. A strong effect against the dengue virus serotype 2 (DENV-2) was also observed suggesting the possibility of treating other Flaviviruses. Additionally, it was shown that the treatment did not reduce the production of infectious viruses in insect cells (C6/36) infected with ZIKV, indicating that the activity of this drug is also related to host factors. Importantly, we demonstrated that NTZ treatment in chorionic and cervical cells caused a reduction of infected cells in a dose-dependent manner and decreased viral loads in up to 2 logs. Pre-clinical in vitro testing evidenced excellent therapeutic response of infected chorionic and cervical cells and point to future NTZ activity investigation in ZIKV congenital transmission models with the perspective of possible repurposing of NTZ to treat Zika fever, especially in pregnant women.

Kelleni’s Protocol Including NSAIDs and Nitazoxanide to Early Manage Dengue Virus Disease: A Novel Potential Broad Spectrum Antiviral Protocol

“…Interestingly, they were likewise falsely recommended against earlier to manage COVID-19 20 before later proven safe and even lifesaving in SARS CoV-2, RSV, Influenza and Norwalk viral infections through earlier real-life practice and later clinical trials 1,21-23 as early treatment aborts the progression of these viral infection and abolishes the occurrence of the complications in the vast majority of cases, in not all of them 24 . Notably, nitazoxanide and/or its active metabolite; tizoxanide were previously shown to in-vitro inhibit the replication of some flavivirdae including dengue virus serotype 2 25,26 . Moreover, nitazoxanide was also suggested to be effective against paramyxovirus infection including parainfluenza and humanmetapneumovirus 27 .…”

Vertical Zika Virus Transmission at the Maternal-Fetal Interface

“…Subsequently, Li et al (146) demonstrated that Nitazoxanide inhibited ZIKV infection by decreasing viral replication and viral protein expression in human placental epithelial cells, human neuronal progenitor cells and human pluripotent stem cell line. Thereafter, De Souza et al (142) found that Nitazoxanide reduced ZIKV viral loads up to 2 logs in primary cultured chorionic cells obtained from human term placentas and in a human cervical epithelial cell line. Recently, Guzeloglu-Kayisli et al ( 104) evaluated the anti-ZIKV activity of tizoxanide in primary cultures of HESCs obtained from cycling…”

Kelleni’s Protocol including Nitazoxanide and NSAIDs to Manage Dengue Virus Disease: A Novel Potential Game-Changer.

“…Notably, nitazoxanide and/or its active metabolite; tizoxanide were previously shown to in-vitro inhibit the replication of some flavivirdae including dengue virus serotype 2 [18,19]. However, until other reports or clinical trials can confirm my recommendation for the early use of ibuprofen/NSAIDs in the management of dengue fever, it is vigilant to prescribe NSAIDs basing on a personalized approach and to prescribe paracetamol together with nitazoxanide to those presenting relatively late while seeking the medical advice and/or lacking urgent medical intervention in their local areas as in the remote impoverished areas, regardless of the results of tourniquet test.…”