TY - JOUR AB - Davoudi-Monfared et al. (1) report in this Journal the results from a clinical trial on COVID-19 patients showing that subcutaneous administration of interferon-β (IFN-β) was associated with a more rapid recovery from SARS-CoV-2 infection and decreased mortality.…. AU - Dorgham, K.* AU - Neumann, A.U. AU - Decavele, M.* AU - Luyt, C.E.* AU - Yssel, H.* AU - Gorochov, G.* C1 - 61328 C2 - 50155 CY - 1752 N St Nw, Washington, Dc 20036-2904 Usa TI - Considering personalized Interferon-β therapy for COVID-19. JO - Antimicrob. Agents Chemother. VL - 65 IS - 4 PB - Amer Soc Microbiology PY - 2021 SN - 0066-4804 ER - TY - JOUR AB - Multidrug resistance among Gram-negative bacteria is a major global public health threat. Metallo-beta-lactamases (MBLs) target the most widely used antibiotic class, the beta-lactams, including the most recent generation of carbapenems. Interspecies spread renders these enzymes a serious clinical threat, and there are no clinically available inhibitors. We present the crystal structures of IMP-13, a structurally uncharacterized MBL from the Gram-negative bacterium Pseudomonas aeruginosa found in clinical outbreaks globally, and characterize the binding using solution nuclear magnetic resonance spectroscopy and molecular dynamics simulations. The crystal structures of apo IMP-13 and IMP-13 bound to four clinically relevant carbapenem antibiotics (doripenem, ertapenem, imipenem, and meropenem) are presented. Active-site plasticity and the active-site loop, where a tryptophan residue stabilizes the antibiotic core scaffold, are essential to the substrate-binding mechanism. The conserved carbapenem scaffold plays the most significant role in IMP-13 binding, explaining the broad substrate specificity. The observed plasticity and substrate-locking mechanism provide opportunities for rational drug design of novel metallo-beta-lactamase inhibitors, essential in the fight against antibiotic resistance. AU - Softley, C. AU - Zak, K.M.* AU - Bostock, M.J. AU - Fino, R. AU - Zhou, R.X. AU - Kolonko, M. AU - Mejdi-Nitiu, R.* AU - Meyer, H.* AU - Sattler, M. AU - Popowicz, G.M. C1 - 58683 C2 - 48296 CY - 1752 N St Nw, Washington, Dc 20036-2904 Usa TI - Structure and molecular recognition mechanism of IMP-13 metallo-β-lactamase. JO - Antimicrob. Agents Chemother. VL - 64 IS - 6 PB - Amer Soc Microbiology PY - 2020 SN - 0066-4804 ER - TY - JOUR AU - Softley, C. AU - Zak, K.M. AU - Bostock, M.J. AU - Fino, R. AU - Zhou, R.X. AU - Kolonko, M. AU - Mejdi-Nitiu, R.* AU - Meyer, H.* AU - Sattler, M. AU - Popowicz, G.M. C1 - 59478 C2 - 48841 CY - 1752 N St Nw, Washington, Dc 20036-2904 Usa TI - Structure and molecular recognition mechanism of IMP-13 metallo-β-lactamase. (vol 64, e00123-20, 2020). JO - Antimicrob. Agents Chemother. VL - 64 IS - 7 PB - Amer Soc Microbiology PY - 2020 SN - 0066-4804 ER - TY - JOUR AB - Hepatitis B virus (HBV) is a major human pathogen, killing an estimated 887,000 people per year. Therefore, potentially curative therapies are of high importance. Following infection, HBV deposits a covalently closed circular DNA (cccDNA) in the nucleus of infected cells that serves as a transcription template and is not affected by current therapies. HBV core protein allosteric modulators (CpAMs) prevent correct capsid assembly but may also affect early stages of HBV infection. In this study, we aimed to determine the antiviral efficacy of a novel, structurally distinct heteroaryldihydropyrimidine (HAP)-type CpAM, HAP_R01, and investigated whether and how HAP_R01 prevents the establishment of HBV infection. HAP_R01 shows a significant inhibition of cccDNA formation when applied during the first 48 h of HBV infection. Inhibiting cccDNA formation, however, requires >1-log(10)-higher concentrations than inhibition of the assembly of newly forming capsids (half-maximal effective concentration [EC50], 345 to 918 nM versus 26.8 to 43.5 nM, respectively). Biophysical studies using a new method to detect the incoming capsid in de novo infection revealed that HAP_R01 can physically change mature capsids of incoming virus particles and affect particle integrity. Treating purified HBV virions with HAP_R01 reduced their infectivity, highlighting the unique antiviral activity of CpAMs to target the capsid within mature HBV particles. Accordingly, HAP_R01 shows an additive antiviral effect in limiting de novo infection when combined with viral entry inhibitors. In summary, HAP_R01 perturbs capsid integrity of incoming virus particles and reduces their infectivity and thus inhibits cccDNA formation in addition to preventing HBV capsid assembly. AU - Ko, C. AU - Bester, R. AU - Zhou, X.* AU - Xu, Z.* AU - Blossey, C. AU - Sacherl, J. AU - Vondran, F.W.R.* AU - Gao, L.* AU - Protzer, U. C1 - 57210 C2 - 47623 CY - 1752 N St Nw, Washington, Dc 20036-2904 Usa TI - A new role for capsid assembly modulators to target mature hepatitis B virus capsids and prevent virus infection. JO - Antimicrob. Agents Chemother. VL - 64 IS - 1 PB - Amer Soc Microbiology PY - 2019 SN - 0066-4804 ER - TY - JOUR AB - HIV replication assays are important tools for HIV drug discovery efforts. Here, we present a full HIV replication system (EASY-HIT) for the identification and analysis of HIV inhibitors. This technology is based on adherently growing HIV-susceptible cells, with a stable fluorescent reporter gene activated by HIV Tat and Rev. A fluorescence-based assay was designed that measures HIV infection by two parameters relating to the early and the late phases of HIV replication, respectively. Validation of the assay with a panel of nine reference inhibitors yielded effective inhibitory concentrations consistent with published data and allowed discrimination between inhibitors of early and late phases of HIV replication. Finer resolution of the effects of reference drugs on different steps of HIV replication was achieved in secondary time-of-addition assays. The EASY-HIT assay yielded high Z' scores (>0.9) and signal stabilities, confirming its robustness. Screening of the LOPAC(1280) library identified 10 compounds (0.8%), of which eight were known to inhibit HIV, validating the suitability of this assay for screening applications. Studies evaluating anti-HIV activities of natural products with the EASY-HIT technology led to the identification of three novel inhibitory compounds that apparently act at different steps of HIV-1 replication. Furthermore, we demonstrate successful evaluation of plant extracts for HIV-inhibitory activities, suggesting application of this technology for the surveillance of biological extracts with anti-HIV activities. We conclude that the EASY-HIT technology is a versatile tool for the discovery and characterization of HIV inhibitors. AU - Kremb, S. AU - Helfer, M. AU - Heller, W. AU - Hoffmann, D.* AU - Wolff, H. AU - Kleinschmidt, A. AU - Cepok, S.* AU - Hemmer, B.* AU - Durner, J. AU - Brack-Werner, R. C1 - 4842 C2 - 27635 SP - 5257-5268 TI - EASY-HIT: HIV full-replication technology forbroad discovery of multiple classes of HIV inhibitors. JO - Antimicrob. Agents Chemother. VL - 54 IS - 12 PB - American Society for Microbiology PY - 2010 SN - 0066-4804 ER - TY - JOUR AB - Since the emergence of viral resistance of hepatitis B virus (HBV) during treatment is becoming an important issue even with newer drugs, there is a need for alternative treatment options such as, for example, RNA interference (RNAi) technology. While short-term suppression of HBV replication is easily achieved with small interfering RNA oligonucleotides, this is not the case for long-term suppression due to the lack of an optimal vector system. Based on the nonviral scaffold/matrix attachment region (S/MAR)-based vector system pEPI-1, which is free of common side effects and is stably retained as an episome even in the absence of selection, we designed a short hairpin RNA (shRNA) expression vector called pEPI-RNAi for HBV suppression. HBV-replicating HepG2.2.15 cells were transfected with pEPI-RNAi, and the intracellular status of the plasmid was followed by PCR and Southern analysis. HBV replication was measured on the DNA, RNA, and protein level. HBV RNA expression was reduced by almost 85% 3 months posttransfection with pEPI-RNAi. At 8 months posttransfection in the absence of antibiotic selection pressure, the suppression level was still 70% and the vector was retained as an episome. The reduction of total intracellular HBV DNA at this point was 77%, showing a marked suppression of HBV DNA replication. At a comparable level, secretion of viral antigens, as well as progeny HBV virions, was inhibited. The S/MAR-based vector system pEPI-1 allows long-term suppression of HBV replication by the expression of suitable shRNAs. Due to its unique properties compared to commonly used vectors, it provides an interesting option for the treatment of chronically HBV-infected individuals. AU - Jenke, A.C.* AU - Wilhelm, A.D.* AU - Orth, V.* AU - Lipps, H.J.* AU - Protzer, U. AU - Wirth, S.* C1 - 2497 C2 - 25464 SP - 2355-2359 TI - Long-term suppression of hepatitis B virus replication by short hairpin RNA expression using the scaffold/matrix attachment region-based replicating vector system pEPI-1. JO - Antimicrob. Agents Chemother. VL - 52 IS - 7 PB - American Society for Microbiology PY - 2008 SN - 0066-4804 ER -