Characterization of a thienylcarboxamide derivative that inhibits the transactivation functions of cytomegalovirus IE2 and varicella zoster virus IE62

Repurposing the clinically approved calcium antagonist manidipine dihydrochloride as a new early inhibitor of human cytomegalovirus targeting the Immediate-Early 2 (IE2) protein

“…This leads to systemic vasodilation and decrease of blood pressure. As a dihydropiridine, MND structurally differs from the other early-acting inhibitors of HCMV recently identified, which are mainly kinase inhibitors (Arend et al, 2017;Beelontally et al, 2017;Strang, 2017) or thyenilcarboxamide derivatives (Majima et al, 2017). It is therefore tempting to speculate that the specific effect shown by MND on IE2 transactivating properties may not be related to the mechanisms of action proposed for other IE2 inhibitors identified by us (such as the 6-aminoquinolone WC5 and its analogs) (Loregian et al, 2010;Massari et al, 2013;Mercorelli et al, 2014) and others.…”

“…IE2 is in fact an essential transactivator of viral E gene expression, autoregulates its own expression, and modulates and hijacks several cellular pathways to promote HCMV replication. Indeed, in the last year we and others have reported the identification of a number of compounds endowed with early anti-HCMV activity, including some able to interfere with IE2-dependent activities (Arend et al, 2017;Beelontally et al, 2017;Majima et al, 2017;Mercorelli et al, 2016;Strang, 2017). In the present study, we have investigated the anti-HCMV properties and mechanism of antiviral activity of MND, a calcium antagonist approved for the treatment of vascular hypertension (McKeage and Scott, 2004).…”

“…Recently, given the importance of IE2 in the HCMV replicative cycle, we and others have put much effort in identifying small molecules able to interfere with IE2 functions and/or its expression (Beelontally et al, 2017;Gardner et al, 2015;Loregian et al, 2010;Majima et al, 2017;Mercorelli et al, 2014Mercorelli et al, , 2016Strang, 2017). In this regard, we used a cell-based assay specific for the IE2dependent transactivation of a reporter gene (Luganini et al, 2008) to screen a library of approved drugs or bioactive molecules.…”

Roles of GP33, a guinea pig cytomegalovirus-encoded G protein-coupled receptor homolog, in cellular signaling, viral growth and inflammation in vitro and in vivo

“…COS-7 cells (product no. CRL1651, ATCC) in 96-well plates were cultured in Dulbecco’s modified medium supplemented with 10% FBS and penicillin/streptomycin, and transiently transfected with a reporter plasmid [pCRE-luc, pNFAT-luc, or pNFκB-luc (Clontech)], an internal control plasmid (pRL-EF1α) [47], and an effector plasmid (pcDNA-GP33s and -GP33) or positive control plasmids, by adding 200-fold diluted lipofectamine 2000 (Invitrogen) and cultured for 6 h. Total amount of DNA for each condition was adjusted by adding the empty vector. The cells were then cultured in a serum-free medium for 24 h and rinsed with PBS.…”