ABSTRACT
Soraphen A is a myxobacterial metabolite that blocks the acetyl-coenzyme A carboxylase of the host and was previously identified as a novel HIV inhibitor. Here, we report that soraphen A acts by reducing virus production and altering the gp120 virion content, impacting entry capacity and infectivity. These effects are partially reversed by addition of palmitic acid, suggesting that inhibition of HIV envelope palmitoylation is one of the mechanisms of antiviral action.
Subject(s)
Anti-HIV Agents/pharmacology , HIV Infections/drug therapy , HIV-1/drug effects , Macrolides/pharmacology , Virus Internalization/drug effects , Virus Replication/drug effects , Acetyl-CoA Carboxylase/antagonists & inhibitors , Cell Line, Tumor , HIV Envelope Protein gp120/metabolism , Humans , Hydroxamic Acids/pharmacology , Lipoylation/drug effects , Myxococcales/metabolism , Palmitic Acid/metabolism , Palmitic Acid/pharmacology , VorinostatABSTRACT
West Nile virus (WNV) is an emerging arbovirus first recognized in Europe in the 1950s. Since then, outbreaks have been reported in several European countries. In 2010, the first WNV outbreak was recorded in Spain, affecting the southern part of the country. We conducted a seroprevalence study in the Catalonia region (northeastern Spain), an area considered at high risk of arbovirus transmission. A total of 800 serum samples from blood donors were collected and screened for antibodies against WNV by enzyme-linked immunosorbent assay (ELISA) and confirmed by a microneutralization assay. More than 50 samples tested positive by ELISA, but only one sample contained neutralizing antibodies against WNV and was obtained from a donor native of Pakistan. The low seroprevalence detected may serve as reference baseline data for monitoring WNV activity in our region in future years.
Subject(s)
Antibodies, Viral/blood , Blood Donors , West Nile Fever/epidemiology , West Nile virus/immunology , Adolescent , Adult , Aged , Antibodies, Neutralizing , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Middle Aged , Seroepidemiologic Studies , Spain/epidemiology , West Nile virus/isolation & purification , Young AdultABSTRACT
BACKGROUND: The nuclear export of unspliced and partially spliced HIV-1 mRNA is mediated by the recognition of a leucine-rich nuclear export signal (NES) in the HIV Rev protein by the host protein CRM1/Exportin1. This makes the CRM1-Rev complex an attractive target for the development of new antiviral drugs. Here we tested the anti-HIV efficacy of ratjadone A, a CRM1 inhibitor derived from myxobacteria. RESULTS: Ratjadone A inhibits HIV infection in vitro in a dose-dependent manner with EC50 values at the nanomolar range. The inhibitory effect of ratjadone A occurs around 12 hours post-infection and is specific for the Rev/CRM1-mediated nuclear export pathway. By using a drug affinity responsive target stability (DARTS) assay we could demonstrate that ratjadone A interferes with the formation of the CRM1-Rev-NES complex by binding to CRM1 but not to Rev. CONCLUSION: Ratjadone A exhibits strong anti-HIV activity but low selectivity due to toxic effects. Although this limits its potential use as a therapeutic drug, further studies with derivatives of ratjadones might help to overcome these difficulties in the future.
Subject(s)
HIV Infections/prevention & control , HIV-1/metabolism , Karyopherins/metabolism , Myxococcales/metabolism , Pyrones/metabolism , Receptors, Cytoplasmic and Nuclear/metabolism , rev Gene Products, Human Immunodeficiency Virus/metabolism , Active Transport, Cell Nucleus/drug effects , Antiviral Agents/pharmacology , Cell Line , HIV Core Protein p24/metabolism , Humans , Karyopherins/antagonists & inhibitors , Protein Binding , Pyrones/chemistry , Pyrones/pharmacology , RNA, Messenger/metabolism , Receptors, Cytoplasmic and Nuclear/antagonists & inhibitors , rev Gene Products, Human Immunodeficiency Virus/antagonists & inhibitors , Exportin 1 ProteinABSTRACT
BACKGROUND: Drug-resistance and therapy failure due to drug-drug interactions are the main challenges in current treatment against Human Immunodeficiency Virus (HIV) infection. As such, there is a continuous need for the development of new and more potent anti-HIV drugs. Here we established a high-throughput screen based on the highly permissive TZM-bl cell line to identify novel HIV inhibitors. The assay allows discriminating compounds acting on early and/or late steps of the HIV replication cycle. RESULTS: The platform was used to screen a unique library of secondary metabolites derived from myxobacteria. Several hits with good anti-HIV profiles were identified. Five of the initial hits were tested for their antiviral potency. Four myxobacterial compounds, sulfangolid C, soraphen F, epothilon D and spirangien B, showed EC50 values in the nM range with SI > 15. Interestingly, we found a high amount of overlapping hits compared with a previous screen for Hepatitis C Virus (HCV) using the same library. CONCLUSION: The unique structures and mode-of-actions of these natural compounds make myxobacteria an attractive source of chemicals for the development of broad-spectrum antivirals. Further biological and structural studies of our initial hits might help recognize smaller drug-like derivatives that in turn could be synthesized and further optimized.
