Anti-HSV-1 synthetic azasteroids display immunomodulating properties

MARÍA E DÁVOLA; JAVIER A. RAMIREZ; LAURA ALCHÉ; ANDREA BARQUERO

Congreso; 34th Annual Meeting of the American Society for Virology; 2015

Herpes simplex virus type 1 (HSV-1)-induced disease occurs as a resultof a primary infection in the epithelium and then, cells like macrophagesintervene in clearing the virus from the infected area and in the developmentof the immunopathological reaction. HSV-1 induces a strong NF-κB nucleartranslocation in different cell lines that could have several functions: topromote viral replication and mediate the cytokine production. For thetreatment of this immunopathology, we have proposed to obtain compounds thatconjugate both antiviral and immunomodulatory activities in the same molecule. Recently, we have synthesized azasteroids with different diamide sidechains; in particular, U20-12 (N-((tert-butylcarbamoyl)methyl)-N-(4-fluorophenyl)-3bhydroxyandrost-5-ene-17b-carboxamide)and U16-L3H (N-((tert-butylcarbamoyl)methyl)-N-(4-chlorophenyl)-3β-hydroxy-20-oxo-pregn-5-ene-16α-carboxamide)have interesting antiherpetic activity invitro. In order to evaluate their potential as future drugs, we exploredthe immunomodulatory properties of both sterol analogues. Therefore, we investigated their effect on NF- κB activation andcytokine secretion induced by viral (HSV-1) and non-viral (LPS or TNF-α)stimuli, in epithelial cells (A549) and macrophages (J774.A1) using indirectimmunofluorescence (IFI) assay and ELISA, respectively. First, we observed that neither U20-12 nor U16-L3H affected the NF-κBnuclear translocation in A549 and J774.A1 cells infected with HSV-1 (moi = 10).Besides, none of them restrained NF-κB translocation in TNF-α-induced epithelialcells and in LPS-stimulated macrophages.Then, we found that U16-L3H significantly increased the production ofIL-8 and IL-6 in comparison with untreated-A549 cells, whereas U20-12 did notaffect the synthesis of either cytokine. In contrast, both compoundssignificantly reduced the secretion of IL-6 and TNF-α in comparison withuntreated-J744.A1 cells. Also, when compounds were added to HSV-1 or LPSstimulated J774.A1, both compounds significantly reduced the production of IL-6and TNF-α in comparison with stimulated-J744.A1 cells.These interesting immunomodulatory properties of U20-12 and U16-L3H,together with their previously reported antiviral activity, make them potentialdrugs for the treatment of HSV-1.