Value through Innovation23 July 2014

Scientific Publications

Boehringer Ingelheim offers an overview of scientific publications on the following pages. This overview represents all publications of the last three years (YTD) where employees of Boehringer Ingelheim worldwide were involved.

73 publications regarding Virology
  • Author:
    Caspari K; Henning H; Schreiber F; Maass P; Gössl R; Schaller C; Waberski D
    Title:
    Impact of porcine circovirus type 2 (PCV2) vaccination on boar semen quality and quantity using two different vaccines
    Source:
    Theriogenology Article in Press (2014)
    Abstract:
    Porcine circovirus type-2 (PCV2) is widespread in domestic pig populations. It can be shed with boar semen, but the role boars have in epidemiology is still unclear. Vaccinating boars against PCV2 can reduce disease and virus load in semen, but may have unwanted side effects, that is, impairment of spermatogenesis. Therefore, the aim of this study was to investigate the effect and impact of two different PCV2 vaccines on boar semen quality and quantity. Healthy normospermic Large White boars in three groups of 12 each were vaccinated with either Circovac, Ingelvac CircoFLEX, or received NaCl. Eight ejaculates were collected starting 1 week after vaccination and assessed for quantitative traits. In general, sperm quantity and quality parameters did not change due to the vaccination (P > 0.05). Only DNA integrity between the Circovac and control group was P < 0.05 but remained at a low level (<2%). One boar showed clinical signs with body temperature up to 39.9 °C and went off feed. For this animal, a clear relation between vaccination, fever period, and impaired sperm quality could be observed. The results indicate that both vaccines did not have a major impact on sperm quality or quantity. Therefore, vaccination of boars against PCV2 seems to be feasible. However, one boar treated with the oil-based vaccine showed a temporarily impaired semen quality after elevated body temperature after vaccination. Thus, possible systemic reactions and the subsequent impact on sperm quality should be taken into account when choosing a PCV2 vaccine for boars. © 2014 Elsevier Inc. All rights reserved.
  • Author:
    Arasteh K; Drulak M; Guo J; Livrozet J-M; Orkin C; Quinson A-M; Ward D
    Title:
    TRANxITION 144 week results: Switching virologically stable HIV patients from immediate-release nevirapine (NVP IR) to extended-release NVP (XR)
    Source:
    J AIDS Clin Res % ($) art. no. 1000292 (2014)
    Abstract:
    Purpose: TRANxITION compared efficacy and safety of switching virologically suppressed HIV-1 infected patients from nevirapine (NVP) immediate-release (IR) (200 mg twice-daily) to NVP extended-release (XR) (400 mg oncedaily). Methods: TRANxITION was an open-label, parallel-group, non-inferiority clinical trial where adult patients with undetectable viral loads, receiving NVP IR plus a fixed-dose NRTI combination, were randomized (2:1) to NVP XR or IR. After week 48, patients initially randomized to IR were allowed to switch to XR. Summary of results: At week 48, proportions of patients with virologic response (LLOQ=50 copies/mL TaqMan, FAS) were 88.5% (131/148) in the IR arm, and 88.8% (262/295) in the XR arm, observed difference of 0.3% (95% CI -6.1, 6.7). DAIDS grade 3/4 adverse events were similar in the XR and IR arms at week 48: 6.4% (19/295) vs. 6.1% (9/148). After week 48, all but 13 patients in the IR arm switched to XR. At week 144, the proportions of patients with virologic response were 95.0% (115/121) in those switching from IR to XR after week 48 [IR-XR post48], and 95.2% (238/250) in those on XR throughout [XR-XR post48]). DAIDS grade 3 and 4 events at week 144 in IR-XRpost48 (7.7%, [10/130]) differed from the XR-XR post48 group (11.2%, [31/276]). Conclusions: NVP XR QD resulted in continued virologic suppression at weeks 48 and 144. Up to week 144, rates of serious AEs were modestly higher than at week 24 in both post-week 48 XR arms, most likely due to the openlabel design of the study. © 2014 Arasteh K, et al.
  • Author:
    Barrier AC; Coombs TM; Dwyer CM; Haskell MJ; Goby L
    Title:
    Administration of a NSAID (meloxicam) affects lying behaviour after caesarean section in beef cows
    Source:
    Appl Anim Behav Sci 155, 28-33 (2014)
    Abstract:
    Very little is known about the pain experienced by cattle following caesarean section and hence little consideration has been given to managing it. Thus the purpose of this study was to investigate activity-related behavioural changes in response to pain, by pre-emptively administering a long acting NSAID to beef cows undergoing caesarean section. One hundred and ten beef cows (55 primiparous, 55 multiparous) that underwent non-elective standardised caesarean section were recruited from eight French veterinary practices. Cows received pre-emptively either meloxicam (n= 63) or placebo (n= 47) according to a blind randomised schedule. Pedometers were attached to each cow's left hindleg and activity was monitored from 0. h (end of surgery) to 68. h post-partum. Time spent lying, number of steps and counts of lying bouts were calculated for the following periods: 0-8. h, 8-16. h, 16-24. h, 24-48. h and 48-68. h and analysed with a REML procedure.Cows receiving meloxicam spent significantly more time lying in the 0-8. h and 8-16. h periods following surgery than cows receiving placebo (+27.4 and +26.4. min, respectively; P<. 0.05) and had more bouts of lying in the first 24. h (P<. 0.05). No differences were seen in the number of steps taken (P>. 0.05). These findings may be interpreted as suggesting that increased lying following caesarean section is an indicator of increased comfort; moreover, it may also suggest that improved welfare, for parturient cows, can be obtained with NSAID treatment. © 2014 Elsevier B.V.
  • Author:
    .krnjug I; Rueckert C; Libanova R; Lienenklaus S; Weiss S; Guzmán CA
    Title:
    The mucosal adjuvant cyclic di-AMP exerts immune stimulatory effects on dendritic cells and macrophages
    Source:
    PLoS ONE 9 (4) art.no.e95728 (2014)
    Abstract:
    The cyclic di-nucleotide bis-(3.,5.)-cyclic dimeric adenosine monophosphate (c-di-AMP) is a candidate mucosal adjuvant with proven efficacy in preclinical models. It was shown to promote specific humoral and cellular immune responses following mucosal administration. To date, there is only fragmentary knowledge on the cellular and molecular mode of action of c-di-AMP. Here, we report on the identification of dendritic cells and macrophages as target cells of c-di-AMP. We show that c-di-AMP induces the cell surface up-regulation of T cell co-stimulatory molecules as well as the production of interferon-.. Those responses were characterized by in vitro experiments with murine and human immune cells and in vivo studies in mice. Analyses of dendritic cell subsets revealed conventional dendritic cells as principal responders to stimulation by c-di-AMP. We discuss the impact of the reported antigen presenting cell activation on the previously observed adjuvant effects of c-di-AMP in mouse immunization studies. © 2014 .krnjug et al.
  • Author:
    Endsley MA; Somasunderam AD; Li G Oezguen N; Thiviyanathan V; Murray JL; Rubin DH; Hodge TW; O'Brien WA; Lewis B; Ferguson MR
    Title:
    Nuclear trafficking of the HIV-1 pre-integration complex depends on the ADAM10 intracellular domain
    Source:
    Virology 454 (455 (1)), 60-66 (2014)
    Abstract:
    Previously, we showed that ADAM10 is necessary for HIV-1 replication in primary human macrophages and immortalized cell lines. Silencing ADAM10 expression interrupted the HIV-1 life cycle prior to nuclear translocation of viral cDNA. Furthermore, our data indicated that HIV-1 replication depends on the expression of ADAM15 and .-secretase, which proteolytically processes ADAM10. Silencing ADAM15 or .-secretase expression inhibits HIV-1 replication between reverse transcription and nuclear entry. Here, we show that ADAM10 expression also supports replication in CD4+ T lymphocytes. The intracellular domain (ICD) of ADAM10 associates with the HIV-1 pre-integration complex (PIC) in the cytoplasm and immunoprecipitates and co-localizes with HIV-1 integrase, a key component of PIC. Taken together, our data support a model whereby ADAM15/.-secretase processing of ADAM10 releases the ICD, which then incorporates into HIV-1 PIC to facilitate nuclear trafficking. Thus, these studies suggest ADAM10 as a novel therapeutic target for inhibiting HIV-1 prior to nuclear entry. © 2014 Elsevier Inc.
  • Author:
    Jeong J; Aly SS; Cano JP; Polson D; Kass PH; Perez AM
    Title:
    Stochastic model of porcine reproductive and respiratory syndrome virus control strategies on a swine farm in the United States
    Source:
    Am J Vet Res 75 (3), 260-267 (2014)
    Abstract:
    Objective-To use mathematical modeling to assess the effectiveness of control strategies for porcine reproductive and respiratory syndrome (PRRS) virus on a swine farm. Sample-A hypothetical small, medium, or large farrow-to-weaning swine farm in the Midwestern United States. Procedures-Stochastic models were formulated to simulate an outbreak of PRRS on a farm. Control strategies assessed in those models included none (baseline) and various combinations of mass immunization, herd closure, and gilt acclimatization. Nine different models resulting from the combination of low, moderate, or high PRRS virus virulence and small, medium, or large herd size were simulated. A stabilized status, the outcome of interest, was defined as the absence of positive PCR assay results for PRRS virus in 3-week-old piglets. For each scenario, the percentage of simulations with a stabilized status was used as a proxy for the probability of disease control. Results-Increasing PRRS virus virulence and herd size were negatively associated with the probability of achieving a stabilized status. Repeated mass immunization with herd closure or gilt acclimitization was a better alternative than was single mass immunization for disease control within a farm. Conclusions and Clinical Relevance-Repeated mass immunization with a PRRS modified- live virus vaccine with herd closure or gilt acclimitization was the scenario most likely to achieve a stabilized status. Estimation of the cost of various PRRS control strategies is necessary.
  • Author:
    LaPlante SR; Padyana AK; Abeywardane A; Bonneau P; Cartier M; Coulombe R; Jakalian A; Wildeson-Jones J; Li X; Liang S; McKercher G; White P; Zhang Q; Taylor SJ
    Title:
    Integrated strategies for identifying leads that target the ns3 helicase of the hepatitis C virus
    Source:
    J Med Chem 57 (5), 2074-2090 (2014)
    Abstract:
    Future treatments for individuals infected by the hepatitis C virus (HCV) will likely involve combinations of compounds that inhibit multiple viral targets. The helicase of HCV is an attractive target with no known drug candidates in clinical trials. Herein we describe an integrated strategy for identifying fragment inhibitors using structural and biophysical techniques. Based on an X-ray structure of apo HCV helicase and in silico and bioinformatic analyses of HCV variants, we identified that one site in particular (labeled 3 + 4) was the most conserved and attractive pocket to target for a drug discovery campaign. Compounds from multiple sources were screened to identify inhibitors or binders to this site, and enzymatic and biophysical assays (NMR and SPR) were used to triage the most promising ligands for 3D structure determination by X-ray crystallography. Medicinal chemistry and biophysical evaluations focused on exploring the most promising lead series. The strategies employed here can have general utility in drug discovery. © 2014 American Chemical Society.
  • Author:
    LaPlante SR; Bös M; Brochu C; Chabot C; Coulombe R; Gillard JR; Jakalian A; Poirier M; Rancourt J; Stammers T; Thavonekham B; Beaulieu PL; Kukolj G; Tsantrizos YS
    Title:
    Conformation-based restrictions and scaffold replacements in the design of hepatitis C virus polymerase inhibitors: Discovery of deleobuvir (BI 207127)
    Source:
    J Med Chem 57 (5), 1845-1854 (2014)
    Abstract:
    Conformational restrictions of flexible torsion angles were used to guide the identification of new chemotypes of HCV NS5B inhibitors. Sites for rigidification were based on an acquired conformational understanding of compound binding requirements and the roles of substituents in the free and bound states. Chemical bioisosteres of amide bonds were explored to improve cell-based potency. Examples are shown, including the design concept that led to the discovery of the phase III clinical candidate deleobuvir (BI 207127). The structure-based strategies employed have general utility in drug design. © 2013 American Chemical Society.
  • Author:
    Jeong J; Aly SS; Cano JP; Polson D; Kass PH; Perez AM
    Title:
    Stochastic model of porcine reproductive and respiratory syndrome virus control strategies on a swine farm in the United States
    Source:
    Am J Vet Res 75 (3), 260-267 (2014)
    Abstract:
    no Abstract available
  • Author:
    Endsley MA; Somasunderam AD; Li G; Oezguen N; Thiviyanathan V; Murray JL; Rubin DH; Hodge TW; O'Brien WA; Lewis B; Ferguson MR
    Title:
    Nuclear trafficking of the HIV-1 pre-integration complex depends on the ADAM10 intracellular domain
    Source:
    Virology 454 (455) (1), 60-66 (2014)
    Abstract:
    no Abstract available
  • Author:
    Berger KL; Triki I; Cartier M; Marquis M; Massariol M-J; Böcher WO; Datsenko Y; Steinmann G; Scherer J; Stern JO; Kukolj G
    Title:
    Baseline hepatitis C virus (HCV) NS3 polymorphisms and their impact on treatment response in clinical studies of the HCV NS3 protease inhibitor faldaprevir
    Source:
    Antimicrob Agents Chemother 58 (2), 698-705 (2014)
    Abstract:
    A challenge to the treatment of chronic hepatitis C with direct-acting antivirals is the emergence of drug-resistant hepatitis C virus (HCV) variants. HCV with preexisting polymorphisms that are associated with resistance to NS3/4A protease inhibitors have been detected in patients with chronic hepatitis C. We performed a comprehensive pooled analysis from phase 1b and phase 2 clinical studies of the HCV protease inhibitor faldaprevir to assess the population frequency of baseline protease inhibitor resistance-associated NS3 polymorphisms and their impact on response to faldaprevir treatment. A total of 980 baseline NS3 sequences were obtained (543 genotype 1b and 437 genotype 1a sequences). Substitutions associated with faldaprevir resistance (at amino acid positions 155 and 168) were rare (<1% of sequences) and did not compromise treatment response: in a phase 2 study in treatment-naive patients, six patients had faldaprevir resistance-associated polymorphisms at baseline, of whom five completed faldaprevir-based treatment and all five achieved a sustained virologic response 24 weeks after the end of treatment (SVR24). Among 13 clinically relevant amino acid positions associated with HCV protease resistance, the greatest heterogeneity was seen at NS3 codons 132 and 170 in genotype 1b, and the most common baseline substitution in genotype 1a was Q80K (99/437 [23%]). The presence of the Q80K variant did not reduce response rates to faldaprevir-based treatment. Across the three phase 2 studies, there was no significant difference in SVR24 rates between patients with genotype 1a Q80K HCV and those without Q80K HCV, whether treatment experienced (17% compared to 26%; P = 0.47) or treatment naive (62% compared to 66%; P = 0.72). Copyright © 2014, American Society for Microbiology. All Rights Reserved.
  • Author:
    Gingras R; Mekhssian K; Fenwick C; White PW; Thibeault D
    Title:
    Human rhinovirus VPg uridylylation alphascreen for high-throughput screening
    Source:
    J Biomol Screen. 19 (2), 259-269 (2014)
    Abstract:
    As an obligate step for picornaviruses to replicate their genome, the small viral peptide VPg must first be specifically conjugated with uridine nucleotides at a conserved tyrosine hydroxyl group. The resulting VPg-pUpU serves as the primer for genome replication. The uridylylation reaction requires the coordinated activity of many components, including the viral polymerase, a conserved internal RNA stem loop structure, and additional viral proteins. Formation of this complex and the resulting conjugation reaction catalyzed by the polymerase, offers a number of biochemical targets for inhibition of an essential process in the viral life cycle. Therefore, an assay recapitulating uridylylation would provide multiple opportunities for discovering potential antiviral agents. Our goal was to identify inhibitors of human rhinovirus (HRV) VPg uridylylation, which might ultimately be useful to reduce or prevent HRV-induced lower airway immunologic inflammatory responses, a major cause of asthma and chronic obstructive pulmonary disease exacerbations. We have reconstituted the complex uridylylation reaction in an AlphaScreen suitable for high-throughput screening, in which a rabbit polyclonal antiserum specific for uridylylated VPg serves as a key reagent. Assay results were validated by quantitative mass spectrometric detection of uridylylation. © 2013 Society for Laboratory Automation and Screening.
  • Author:
    Stephan C; León W
    Title:
    Finding of nevirapine extended release tablet remnants in stools does not threaten the success of combination antiretroviral therapy
    Source:
    HIV Med 15 (2), 124-128 (2014)
    Abstract:
    Objectives: The finding of nevirapine extended release (XR) tablet remnants in stools has raised concerns about emerging HIV-1 resistance. The aim of this study was to evaluate the characteristics and pharmacokinetic and virological outcomes of affected patients from clinical trials. Methods: HIV-1-infected individuals reporting tablet remnants in stools during phase III (VERxVE and TRANxITION-studies)-clinical trials were evaluated for mean pharmacokinetic nevirapine concentrations in available blood trough samples and remnants from stool. Patient characteristics including age, race, geographical region and primary study endpoint outcome were investigated for risk factor association with the finding of tablet remnants. Results: Of 800 patients receiving the nevirapine XR formulation, 15 reported tablet remnants in stools, an incidence rate of 1.19% in VERxVE and 3.05% in the TRANxITION study. The difference in event rate was highly significant between the XR and immediate release (IR) formulations (P<0.001), but not between trials (P=0.061). All patients (15 of 15) reporting remnants achieved the primary study endpoint of HIV-1 suppression (<50 HIV-1 RNA copies/mL), whereas overall 81% of patients in the VERxVE trial and 94% in the TRANxITION trial did so. The mean nevirapine trough concentration was 3431.4ng/mL in patients reporting remnants. Tablet remnants retrieved from the stools of three subjects revealed a percentage nevirapine recovery of 22.8-42.2% of original drug. Subgroup analysis of gender, age, race and geographical region revealed no risk factor association with the finding of remnants. Conclusions: The finding of nevirapine tablet remnants in stools is a rare event, with an incidence of approximately 2%, restricted to the XR formulation. Affected patients responded fully to antiretroviral therapy by achieving the primary study endpoint and demonstrating no relevant safety risks; nevirapine pharmacokinetic analysis of blood and stool samples ruled out underexposure. © 2013 British HIV Association.
  • Author:
    Stammers TA; Coulombe R; Duplessis M; Fazal G; Gagnon A; Garneau M; Goulet S; Jakalian A; Laplante S; Rancourt J; Thavonekham B; Wernic D; Kukolj G; Beaulieu PL
    Title:
    Anthranilic acid-based Thumb Pocket 2 HCV NS5B polymerase inhibitors with sub-micromolar potency in the cell-based replicon assay
    Source:
    Bioorg Med Chem Lett 23 (24), 6879-6885 (2013)
    Abstract:
    Optimization efforts on the anthranilic acid-based Thumb Pocket 2 HCV NS5B polymerase inhibitors 1 and 2 resulted in the identification of multiple structural elements that contributed to improved cell culture potency. The additive effect of these elements resulted in compound 46, an inhibitor with enzymatic (IC50) and cell culture (EC50) potencies of less than 100 nanomolar. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Rey MR; Undi M; Rodriguez-Lecompte JC; Joseph T; Morrison J; Yitbarek A; Wittenberg K; Tremblay R; Crow GH; Ominski KH
    Title:
    A study of the effectiveness of a needle-free injection device compared with a needle and syringe used to vaccinate calves against bovine viral diarrhea and infectious bovine rhinotracheitis viruses
    Source:
    Vet J (Lond) 198 (1), 235-238 (2013)
    Abstract:
    The aim of this study was to compare the effectiveness of a needle-free injection device (NF) with a needle and syringe (NS) when used to vaccinate calves against bovine viral diarrhea virus (BVDV) and infectious bovine rhinotracheitis virus (IBRV). The study was conducted in two independent phases. Ninety-six crossbred beef calves were vaccinated in the spring and 98 beef calves in the autumn. The calves were vaccinated using a NF or NS at 2. months of age (day 0) and again on day 119, with a modified-live virus vaccine containing IBRV, BVDV (types 1 and 2), parainfluenza-3 virus, and bovine respiratory syncytial virus. In each herd 10 calves were left unvaccinated to determine whether exposure to either BVDV or IBRV occurred. Visible vaccine residue at the surface of the skin/hair was apparent immediately following vaccination with NF in 30% of the spring-born calves following both the primary and booster vaccination. In the autumn, visible vaccine residues occurred in 19% and 8% of NF-vaccinated calves following the primary and booster vaccination. Post-vaccination skin reactions recorded on days 21, 42, 119 and 140 occurred with greater frequency in NF-vaccinated calves than NS-vaccinated ones. Blood samples were collected on days 0, 21, 42, 119, and 140 and tested for antibodies to BVDV and IBRV. Vaccination technique had no significant effect on BVDV or IBRV antibody concentrations at any time point. NF was as effective as NS vaccination in eliciting BVDV and IBRV antibody responses. © 2013 Elsevier Ltd.
  • Author:
    Beato MS; Realpe-Quintero M; Bonfante F; Mancin M; Ormelli S; Terregino C; Gonzalez-Hernandez C; Capua In B
    Title:
    Cross-clade protection against H5N1 HPAI strains recently isolated from commercial poultry in Egypt with a single dose of a baculovirus based vaccine
    Source:
    Vaccine 31 (44), 5075-5081 (2013)
    Abstract:
    The current avian influenza epidemic in Egypt caused by circulation of genetically and antigenically diverse H5N1 HPAI viruses in poultry is controlled by applying vaccination among other measures. In this context, the use of a DIVA (differentiating infected from vaccinated animals) vaccination strategy utilizing a vaccine capable of inducing protection against multiple antigenic variants may result as an additional control tool to the existing ones. In this study the efficacy of a single-shot recombinant baculovirus-based vaccine in specific-pathogen-free chickens was tested by experimental challenge with genetically and antigenically diverse H5N1 HPAI viruses belonging to clades 2.2.1 and 2.2.1.1, which have been circulating in Egypt since 2010. A single dose of vaccine, administration at 10 days of age, was shown to confer 100% clinical protection, with a decrease or suppression of virus shedding. © 2013 Elsevier Ltd.
  • Author:
    Escobar PA; Kemper RA; Tarca J; Nicolette J; Kenyon M; Glowienke S; Sawant SG; Christensen J; Johnson TE; McKnight C; Ward G; Galloway SM; Custer L; Gocke E; O'Donovan MR; Braun K; Snyder RD; Mahadeva
    Title:
    Corrigendum to "Bacterial mutagenicity screening in the pharmaceutical industry
    Source:
    Mutat Res Rev Mutat Res 752 (2), 99-118 Article in press (2013)
    Abstract:
    no abstract available
  • Author:
    Parzych EM; Dimenna LJ; Latimer BP; Small JC; Kannan S; Manson B; Lasaro MO; Wherry EJ; Ertl HC
    Title:
    Influenza virus specific CD8+ T cells exacerbate infection following high dose influenza challenge of aged mice
    Source:
    BioMed Res Int art.no. 876314 (2013)
    Abstract:
    Influenza viruses cause severe illnesses and death, mainly in the aged population. Protection afforded by licensed vaccines through subtype-specific neutralizing antibodies is incomplete, especially when the vaccine antigens fail to closely match those of the circulating viral strains. Efforts are underway to generate a so-called universal influenza vaccine expressing conserved viral sequences that induce broad protection to multiple strains of influenza virus through the induction of CD8+ T cells. Here we assess the effect of a potent antiviral CD8+ T cell response on influenza virus infection of young and aged mice. Our results show that CD8+ T cell-inducing vaccines can provide some protection to young mice, but they exacerbate influenza virus-associated disease in aged mice, causing extensive lung pathology and death. © 2013 E. M. Parzych et al.
  • Author:
    Kramps T; Probst J
    Title:
    Messenger RNA-based vaccines: Progress, challenges, applications
    Source:
    Wiley Interdiscip Rev Syst Biol Med 4 (6), 737-749 (2013)
    Abstract:
    Twenty years after the demonstration that messenger RNA (mRNA) was expressed and immunogenic upon direct injection in mice, the first successful proof-of-concept of specific protection against viral infection in small and large animals was reported. These data indicate wider applicability to infectious disease and should encourage continued translation of mRNA-based prophylactic vaccines into human clinical trials. At the conceptual level, mRNA-based vaccines-more than other genetic vectors-combine the simplicity, safety, and focused immunogenicity of subunit vaccines with favorable immunological properties of live viral vaccines: (1) mRNA vaccines are molecularly defined and carry no excess information. In the environment and upon physical contact, RNA is rapidly degraded by ubiquitous RNases and cannot persist. These characteristics also guarantee tight control over their immunogenic profile (including avoidance of vector-specific immune responses that could interfere with repeated administration), pharmacokinetics, and dosing. (2) mRNA vaccines are synthetically produced by an enzymatic process, just requiring information about the nucleic acid sequence of the desired antigen. This greatly reduces general complications associated with biological vaccine production, such as handling of infectious agents, genetic variability, environmental risks, or restrictions to vaccine distribution. (3) RNA can be tailored to provide potent adjuvant stimuli to the innate immune system by direct activation of RNA-specific receptors; this may reduce the need for additional adjuvants. The formation of native antigen in situ affords great versatility, including intracellular localization, membrane association, posttranslational modification, supra-molecular assembly, or targeted structural optimization of delivered antigen. Messenger RNA vaccines induce balanced immune responses including B cells, helper T cells, and cytotoxic T lymphocytes, rendering them an extremely adaptable platform. This article surveys the design, mode of action, and capabilities of state-of-the-art mRNA vaccines, focusing on the paradigm of influenza prophylaxis. © 2013 John Wiley & Sons, Ltd.
  • Author:
    Stephan C; León W
    Title:
    Finding of nevirapine extended release tablet remnants in stools does not threaten the success of combination antiretroviral therapy
    Source:
    HIV Med Article in press (2013)
    Abstract:
    Objectives: The finding of nevirapine extended release (XR) tablet remnants in stools has raised concerns about emerging HIV-1 resistance. The aim of this study was to evaluate the characteristics and pharmacokinetic and virological outcomes of affected patients from clinical trials. Methods: HIV-1-infected individuals reporting tablet remnants in stools during phase III (VERxVE and TRANxITION-studies)-clinical trials were evaluated for mean pharmacokinetic nevirapine concentrations in available blood trough samples and remnants from stool. Patient characteristics including age, race, geographical region and primary study endpoint outcome were investigated for risk factor association with the finding of tablet remnants. Results: Of 800 patients receiving the nevirapine XR formulation, 15 reported tablet remnants in stools, an incidence rate of 1.19% in VERxVE and 3.05% in the TRANxITION study. The difference in event rate was highly significant between the XR and immediate release (IR) formulations (P<0.001), but not between trials (P=0.061). All patients (15 of 15) reporting remnants achieved the primary study endpoint of HIV-1 suppression (<50 HIV-1 RNA copies/mL), whereas overall 81% of patients in the VERxVE trial and 94% in the TRANxITION trial did so. The mean nevirapine trough concentration was 3431.4ng/mL in patients reporting remnants. Tablet remnants retrieved from the stools of three subjects revealed a percentage nevirapine recovery of 22.8-42.2% of original drug. Subgroup analysis of gender, age, race and geographical region revealed no risk factor association with the finding of remnants. Conclusions: The finding of nevirapine tablet remnants in stools is a rare event, with an incidence of approximately 2%, restricted to the XR formulation. Affected patients responded fully to antiretroviral therapy by achieving the primary study endpoint and demonstrating no relevant safety risks; nevirapine pharmacokinetic analysis of blood and stool samples ruled out underexposure. © 2013 British HIV Association
  • Author:
    Brinson C; Bogner JR; Nelson M; Podzamczer D; Quinson AM; Drulak M; Andrade-Villanueva J; Cahn P; Santiago S; Gathe J
    Title:
    Verxve 144-week results: Nevirapine extended release (nvp xr) qd versus nvp immediate release (ir) bid with ftc/tdf in treatment-naive hiv-1 patients
    Source:
    J AIDS Clin Res 4 (8) (2013)
    Abstract:
    Background: We report 96- and 144-week follow-up data from VERxVE, demonstrating that nevirapine (NVP) extended release (XR) 400 mg once daily was non-inferior to NVP immediate release (IR) 200 mg twice daily, each administered on a backbone of emtricitabine/tenofovir. Methods: VERxVE was a double-blind, double-dummy, non-inferiority study in adults with screening viral load (VL) &gt;1000 copies/mL and CD4+ cell count &lt;400 cells/mm3 (males) or &lt;250 cells/mm3 (females). Randomisation was stratified by baseline VL (copies/mL) . 100,000 or &gt;100,000. The primary endpoint was confirmed virologic response (&lt;50 copies/mL) at week 48. Cochran's statistic incorporating baseline VL strata tested non-inferiority of XR efficacy versus IR. Secondary endpoints included 144-week sustained virologic response and safety. Results: In all, 1011 patients were randomised and treated with NVP: 736 (XR n=378; IR n= 358) completed 144 weeks. Virologic response was 63.6% for NVP XR and 58.5% for NVP IR (adjusted difference of 4.8% [95% confidence interval -1.1% to 10.8%] favouring NVP XR). No significant differences were seen in changes in CD4+ T-cell counts from baseline, virologic failures or total discontinuation rates between treatment arms, regardless of demographic or baseline characteristics. Conclusions: NVP XR demonstrated non-inferior virologic efficacy to NVP IR in treatment-naïve HIV-infected patients and was well tolerated out to week 144, with a safety profile similar to NVP IR. © 2013 Brinson C, et al.
  • Author:
    Zeuzem S; Mensa F J
    Title:
    Concordance between sustained virologic response week 12 (SVR12) and SVR24 in genotype 1 hepatitis C virus patients receiving interferon-free treatment in the SOUND-C2 study
    Source:
    Hepatology 58 (4), 1516 (2013)
    Abstract:
    no abstract available
  • Author:
    Iyer AV; Kousoulas KG
    Title:
    A review of vaccine approaches for West Nile virus
    Source:
    Int J Environ Res Public Health 10 (9), 4200-4223 (2013)
    Abstract:
    The West Nile virus (WNC) first appeared in North America in 1999. The North American lineages of WNV were characterized by the presence of neuroinvasive and neurovirulent strains causing disease and death in humans, birds and horses. The 2012 WNV season in the United States saw a massive spike in the number of neuroinvasive cases and deaths similar to what was seen in the 2002-2003 season, according to the West Nile virus disease cases and deaths reported to the CDC by year and clinical presentation, 1999-2012, by ArboNET (Arboviral Diseases Branch, Centers for Disease Control and Prevention). In addition, the establishment and recent spread of lineage II WNV virus strains into Western Europe and the presence of neurovirulent and neuroinvasive strains among them is a cause of major concern. This review discusses the advances in the development of vaccines and biologicals to combat human and veterinary West Nile disease. © 2013 by the authors; licensee MDPI, Basel, Switzerland.
  • Author:
    Lamorte L; Titolo S; Lemke CT; Goudreau N; Mercier J-F; Wardrop E; Shah VB; Von Schwedler UK; Langelier C; Banik SSR; Aiken C; Sundquist WI; Mason SW
    Title:
    Discovery of novel small-molecule HIV-1 replication inhibitors that stabilize capsid complexes
    Source:
    Antimicrob Agents Chemother 57 (10), 4622-4631 (2013)
    Abstract:
    The identification of novel antiretroviral agents is required to provide alternative treatment options for HIV-1-infected patients. The screening of a phenotypic cell-based viral replication assay led to the identification of a novel class of 4,5-dihydro-1Hpyrrolo[ 3,4-c]pyrazol-6-one (pyrrolopyrazolone) HIV-1 inhibitors, exemplified by two compounds: BI-1 and BI-2. These compounds inhibited early postentry stages of viral replication at a step(s) following reverse transcription but prior to 2 long terminal repeat (2-LTR) circle formation, suggesting that they may block nuclear targeting of the preintegration complex. Selection of viruses resistant to BI-2 revealed that substitutions at residues A105 and T107 within the capsid (CA) amino-terminal domain (CANTD) conferred high-level resistance to both compounds, implicating CA as the antiviral target. Direct binding of BI-1 and/or BI-2 to CANTD was demonstrated using isothermal titration calorimetry and nuclear magnetic resonance (NMR) chemical shift titration analyses. A high-resolution crystal structure of the BI-1:CANTD complex revealed that the inhibitor bound within a recently identified inhibitor binding pocket (CANTD site 2) between CA helices 4, 5, and 7, on the surface of the CANTD, that also corresponds to the binding site for the host factor CPSF-6. The functional consequences of BI-1 and BI-2 binding differ from previously characterized inhibitors that bind the same site since the BI compounds did not inhibit reverse transcription but stabilized preassembled CA complexes. Hence, this new class of antiviral compounds binds CA and may inhibit viral replication by stabilizing the viral capsid. Copyright © 2013, American Society for Microbiology. All Rights Reserved
  • Author:
    Rey MR; Undi M; Rodriguez-Lecompte JC; Joseph T; Morrison J; Yitbarek A; Wittenberg K; Tremblay R; Crow GH; Ominski KH
    Title:
    A study of the effectiveness of a needle-free injection device compared with a needle and syringe used to vaccinate calves against bovine viral diarrhea and infectious bovine rhinotracheitis viruses
    Source:
    Br Vet J Article in press (2013)
    Abstract:
    The aim of this study was to compare the effectiveness of a needle-free injection device (NF) with a needle and syringe (NS) when used to vaccinate calves against bovine viral diarrhea virus (BVDV) and infectious bovine rhinotracheitis virus (IBRV). The study was conducted in two independent phases. Ninety-six crossbred beef calves were vaccinated in the spring and 98 beef calves in the autumn. The calves were vaccinated using a NF or NS at 2 months of age (day 0) and again on day 119, with a modified-live virus vaccine containing IBRV, BVDV (types 1 and 2), parainfluenza-3 virus, and bovine respiratory syncytial virus. In each herd 10 calves were left unvaccinated to determine whether exposure to either BVDV or IBRV occurred. Visible vaccine residue at the surface of the skin/hair was apparent immediately following vaccination with NF in 30% of the spring-born calves following both the primary and booster vaccination. In the autumn, visible vaccine residues occurred in 19% and 8% of NF-vaccinated calves following the primary and booster vaccination. Post-vaccination skin reactions recorded on days 21, 42, 119 and 140 occurred with greater frequency in NF-vaccinated calves than NS-vaccinated ones. Blood samples were collected on days 0, 21, 42, 119, and 140 and tested for antibodies to BVDV and IBRV. Vaccination technique had no significant effect on BVDV or IBRV antibody concentrations at any time point. NF was as effective as NS vaccination in eliciting BVDV and IBRV antibody responses. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Zeuzem S; Mensa FJ
    Title:
    Concordance between sustained virologic response week 12 (SVR12) and SVR24 in genotype 1 hepatitis C virus patients receiving interferon-free treatment in the SOUND-C2 study
    Source:
    Hepatology Article in press (2013)
    Abstract:
    no abstract available
  • Author:
    Nishiguchi S; Sakai Y; Kuboki M; Tsunematsu S; Urano Y; Sakamoto W; Tsuda Y; Steinmann G; Omata M
    Title:
    Safety and efficacy of faldaprevir with pegylated interferon alfa-2a and ribavirin in Japanese patients with chronic genotype-1 hepatitis C infection
    Source:
    Liver Int Article in press (2013)
    Abstract:
    Abstract: Background & Aims: Faldaprevir (BI 201335) is a potent once-daily (QD) NS3/4A protease inhibitor for the treatment of patients with genotype-1 (GT-1) hepatitis C virus (HCV). The aim of this study was to evaluate the safety, pharmacokinetics and efficacy of faldaprevir plus pegylated interferon alfa-2a (PegIFN) and ribavirin (RBV) in Japanese patients infected with chronic GT-1 HCV. Methods: Part 1 of this phase II study was a randomized, double-blind, placebo-controlled, dose-ascending study. Treatment-naïve patients received faldaprevir 120 or 240 mg QD, or placebo, plus PegIFN/RBV for 4 weeks, then PegIFN/RBV alone for 44 weeks. In Part 2 (open label), treatment-experienced patients received faldaprevir 240 mg QD plus PegIFN/RBV for 4 weeks, then PegIFN/RBV alone for 44 weeks. Efficacy was assessed using sustained virological response (SVR) 24 weeks after treatment completion. The pharmacokinetics, safety and tolerability of faldaprevir were also assessed. Results: SVR was achieved by 4/6 (67%) treatment-naïve patients treated with faldaprevir 120 mg QD, 5/6 (83%) patients treated with faldaprevir 240 mg QD and 2/4 (50%) patients who received placebo. Of the treatment-experienced patients, 3/6 (50%) achieved SVR. Faldaprevir was well tolerated. There was one serious adverse event, which was not considered to be treatment related. Rash and hyperbilirubinaemia were more frequently reported with faldaprevir than with placebo in treatment-naïve patients, but no cases were severe or serious and none led to discontinuation. Steady-state plasma concentrations of faldaprevir were reached within 7 days of QD dosing. Conclusions: Faldaprevir with PegIFN/RBV was efficacious and well tolerated, supporting further evaluation of this combination in Japanese patients. © 2013 John Wiley & Sons A/S.
  • Author:
    Alonso C; Davies PR; Polson DD; Dee SA; Lazarus WF
    Title:
    Financial implications of installing air filtration systems to prevent PRRSV infection in large sow herds
    Source:
    Prev Vet Med 111 (3) (4), 268-277 (2013)
    Abstract:
    Air filtration systems implemented in large sow herds have been demonstrated to decrease the probability of having a porcine reproductive and respiratory syndrome virus (PRRSV) outbreak. However, implementation of air filtration represents a considerable capital investment, and does not eliminate the risk of new virus introductions. The specific objectives of the study were: 1) to determine productivity differences between a cohort of filtered and non-filtered sow farms; and 2) to employ those productivity differences to model the profitability of filtration system investments in a hypothetical 3000 sow farm. Variables included in the study were production variables (quarterly) from respective herds; air filtration status; number of pig sites within 4.7. km of the farm; occurrence of a PRRSV outbreak in a quarter, and season. For the investment analyses, three Scenarios were compared in a deterministic spreadsheet model of weaned pig cost: (1) control, (2) filtered conventional attic, and (3) filtered tunnel ventilation. Model outputs indicated that a filtered farm produced 5927 more pigs than unfiltered farms. The payback periods for the investments, were estimated to be 5.35 years for Scenario 2 and 7.13 years for Scenario 3 based solely on sow herd productivity. Payback period sensitivity analyses were performed for both biological and financial inputs. The payback period was most influenced by the premium for weaned pig sales price for PRRSV-negative pigs, and the relative proportions of time that filtered vs. unfiltered farms produced PRRSV-negative pigs. A premium of $5 per pig for PRRS-negative weaned pigs reduced the estimated payback periods to 2.1 years for Scenario 2 and 2.8 years for Scenario 3. © 2013 Elsevier B.V.
  • Author:
    Bethell R; Scherer J; Witvrouw M; Paquet A; Coakley E; Hall D
    Title:
    Short communication: Phenotypic protease inhibitor resistance and cross-resistance in the clinic from 2006 to 2008 and mutational prevalences in HIV from patients with discordant tipranavir and darunavir susceptibility phenotypes.
    Source:
    AIDS Res Hum Retroviruses 28 (9), 1019-1024 (2012)
    Abstract:
    To test tipranavir (TPV) or darunavir (DRV) as treatment options for patients with phenotypic resistance to protease inhibitors (Pis), including lopinavir, saquinavir, atazanavir, and fosamprenavir, the PhenoSense GT database was analyzed for susceptibility to DRV or TPV among Pl-resistant isolates. The Monogram Bioseiences HIV database (South San Francisco, CA) containing 7775 clinical isolates (2006-2008) not susceptible to at least one first-generation PI was analyzed. Phenotypic responses [resistant (R), partially susceptible (PS), or susceptible (S)] were defined by upper and lower clinical cut-offs to each PI. Genetypes were screened for amino acid substitutions associated with TPV-R/DRV-S and TPV-S/DRV-R phenotypes. ln all, 4.9% (378) of isolates were resistant to all six Pis and 31.0% (2407) were resistant to none. Among isolates resistant to all four first-generation Pis, DRV resistance increased from 21.2% to 41.9% from 2006 to 2008, respectively, and resistance to TPV remained steady (53.9 to 57.3%, respectively). Higher prevalence Substitutions in DRV-S/TPV-R isolates versus DRV-R/TPV-S isolates, respectively, were 82L/T (44.4% vs. 0%) and 830 (5.8% vs. 0%). Higher prevalence Substitutions in DRV-R/TPV-S virus were 50V (0.0% vs. 28.9%), 54L (1.0% vs. 36.1%), and 76V (0.4% vs. 15.5%). Mutations to help predict discordant susceptibility to DRV and TPV in isolates with reduced susceptibility to other Pis were identified. DRV resistance mutations associated with improved virologic response to TPV were more prevalent in DRV-R/TPV-S isolates. TPV resistance mutations were more prevalent in TPV-R and DRV-S isolates. These results confirm the impact of genotype on phenotype, illustrating how HIV genotype and phenotype data assist regimen optimization.
  • Author:
    EI-Far M; Isabelle C; Chomont N; Bourbonniere M; Fonseca S; Ancuta P; Peretz Y; Chouikh Y; Halwani R; SchwartzO; Madrenas J; Freeman GJ; Routy J-P; Haddad EK; Sekaly R-
    Title:
    Down-Regulation of CTLA-4 by HIV-1 Nef Protein
    Source:
    PLoS ONE 8 (1) art no 54295 (2013)
    Abstract:
    HIV-1 Nef protein down-regulates several cell surface receptors through its interference with the cell sorting and trafficking machinery. Here we demonstrate for the firsttime the ability of Nef to down-regulate cell surface expression of the negative immune modulator CTLA-4. Down-regulation of CTLA-4 required the Nef motifs DD175, EE155 and LL165, all known tobe involved in vesicle trafficking. Disruption of the lysosomal functions by pH-neutralizing agents prevented CTLA-4 down-regulation by Nef, demonstrating the implication of the endosomal/lysosomal compartments in this process. Confocal microscopy experiments visualized the co-localization between Nef and CTLA-4 in the early and recycling endosomes but not at the cell surface. Overall, our results provide a novel mechanism by which HIV-1 Nef interferes with the surface expression of the negative regulatorofT cell activation CTLA-4. Down-regulation of CTLA-4 may contribute to the mechanisms by which HIV-1 sustains T cell activation, a critical step in viral replication and dissemination. © 2013 EI-Far et al.
  • Author:
    Wernike K; Nikolin VM; Hechinger S; Hoffmann B; Beer M
    Title:
    Inactivated Schmallenberg virus prototype vaccines
    Source:
    Vaccine 31 (35), 3558-3563 (2013)
    Abstract:
    Schmallenberg virus (SBV), a novel Orthobunyavirus, is an insect-transmitted pathogen and was first described in Europe in 2011. SBV causes a mild transient disease in adult ruminants, but severe foetal malformation and stillbirth were observed after an infection of naive cows and ewes, which is responsible for considerable economic losses. The virus is now widely distributed in Europe, and no vaccines were available to stop transmission and spread. In the present study, 16 calves and 25 sheep, the major target species of SBV infection, were vaccinated twice 3 weeks apart with one of 5 newly developed, inactivated vaccine candidates. Six calves and 5 sheep were kept as unvaccinated controls. All animals were clinically, serologically and virologically examined before and after challenge infection. Immunisation with the inactivated preparations resulted in a neutralising antibody response three weeks after the second vaccination without any side effects. The number of animals that seroconverted in each group and the strength of the antibody response were dependent on the cell line used for virus growth and on the viral titre prior to inactivation. Four vaccine prototypes completely prevented RNAemia after challenge infection, a fifth candidate reduced RNAemia considerably. Although further evaluations e.g. regarding duration of immunity will be necessary, the newly developed vaccines are promising candidates for the prevention of SBV-infection and could be a valuable tool in SBV control strategies. © 2013 Elsevier Ltd.
  • Author:
    El-Far M; Isabelle C; Chomont N; Bourbonnière M; Fonseca S; Ancuta P; Peretz Y; Chouikh Y; Halwani R; Schwartz O; Madrenas J; Freeman GJ; Routy JP; Haddad EK; Sékaly RP
    Title:
    Down-Regulation of CTLA-4 by HIV-1 Nef Protein.
    Source:
    PLoS One 8 (1) e54295 (2013)
    Abstract:
    HIV-1 Nef protein down-regulates several cell surface receptors through its interference with the cell sorting and trafficking machinery. Here we demonstrate for the first time the ability of Nef to down-regulate cell surface expression of the negative immune modulator CTLA-4. Down-regulation of CTLA-4 required the Nef motifs DD175, EE155 and LL165, all known to be involved in vesicle trafficking. Disruption of the lysosomal functions by pH-neutralizing agents prevented CTLA-4 down-regulation by Nef, demonstrating the implication of the endosomal/lysosomal compartments in this process. Confocal microscopy experiments visualized the co-localization between Nef and CTLA-4 in the early and recycling endosomes but not at the cell surface. Overall, our results provide a novel mechanism by which HIV-1 Nef interferes with the surface expression of the negative regulator of T cell activation CTLA-4. Down-regulation of CTLA-4 may contribute to the mechanisms by which HIV-1 sustains T cell activation, a critical step in viral replication and dissemination.
  • Author:
    Moreau Benoit; O'Meara Jeff A; Bordeleau Josee; Garneau Michel; Godbout Cedrickx; Gorys Vida; Leblanc Melissa; Villemure Elisia; White Peter W; Llinas-Brunet Montse
    Title:
    Discovery of Hepatitis C Virus NS3-4A Protease Inhibitors with Improved Barrier to Resistance and Favorable Liver Distribution
    Source:
    J Med Chem (2013)
    Abstract:
    Given the emergence of resistance observed for the current clinical-stage hepatitis C virus (HCV) NS3 protease inhibitors, there is a need for new inhibitors with a higher barrier to resistance. We recently reported our rational approach to the discovery of macrocyclic acylsulfonamides as HCV protease inhibitors addressing potency against clinically relevant resistant variants. Using X-ray crystallography of HCV protease variant/inhibitor complexes, we shed light on the complex structural mechanisms by which the D168V and R155K residue mutations confer resistance to NS3 protease inhibitors. Here, we disclose SAR investigation and ADME/PK optimization leading to the identification of inhibitors with significantly improved potency against the key resistant variants and with increased liver partitioning.
  • Author:
    Bailey MD; Halmos T; Lemke CT
    Title:
    Discovery of novel P2 substituted 4-biaryl proline inhibitors of hepatitis C virus NS3 serine protease
    Source:
    Bioorg Med Chem Lett 23 (15), 4436-4440 (2013)
    Abstract:
    Inhibitors of hepatitis C virus NS3 serine protease often incorporate a large P2 moiety to interact with the surface of the enzyme while shielding part of the catalytic triad. This feature is important in many inhibitors in order to have the necessary potency needed for efficacy. In this Letter we explore some new P2 motifs to further exploit this region of the enzyme. In a continuing effort to replace the often found 4-hydroxyproline P2 core found in the majority of inhibitors for this target, various directly attached aryl derivatives were evaluated. Of these, the 2,4-disubstituted thiazole core proved to be the most interesting. SAR around this motif has lead to compounds with Ki's in the high picomolar range and provided cellular potencies in the single digit nM range. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Jacob CL; Lamorte L; Sepulveda E; Lorenz IC; Gauthier A; Franti
    Title:
    Neutralizing antibodies are unable to inhibit direct viral cell-to-cell spread of human cytomegalovirus
    Source:
    Virology Article in press (2013)
    Abstract:
    Infection with human cytomegalovirus (CMV) during pregnancy is the most common cause of congenital disorders, and can lead to severe life-long disabilities with associated high cost of care. Since there is no vaccine or effective treatment, current efforts are focused on identifying potent neutralizing antibodies. A panel of CMV monoclonal antibodies identified from patent applications, was synthesized and expressed in order to reproduce data from the literature showing that anti-glycoprotein B antibodies neutralized virus entry into all cell types and that anti-pentameric complex antibodies are highly potent in preventing virus entry into epithelial cells. It had not been established whether antibodies could prevent subsequent rounds of infection that are mediated primarily by direct cell-to-cell transmission. A thorough validation of a plaque reduction assay to monitor cell-to-cell spread led to the conclusion that neutralizing antibodies do not significantly inhibit plaque formation or reduce plaque size when they are added post-infection. © 2013 Elsevier Inc. All rights reserved.
  • Author:
    Sulkowski MS; Bourlière M; Bronowicki J-P; Asselah T; Pawlotsky J-M; Shafran SD; Pol S; Mauss S; Larrey D; Datsenko Y; Stern JO; Kukolj G; Scherer J; Nehmiz G; Steinmann GG; Böcher WO
    Title:
    Faldaprevir combined with peginterferon alfa-2a and ribavirin in chronic hepatitis C virus genotype-1 patients with prior nonresponse: SILEN-C2 trial
    Source:
    Hepatology 57 (6), 2155-2163 (2013)
    Abstract:
    Faldaprevir (BI 201335) is a potent, hepatitis C virus (HCV) NS3/4A protease inhibitor. In all, 290 noncirrhotic HCV genotype (GT)-1 patients with prior null (&lt;1 log10 viral load [VL] drop at any time on treatment) or partial response (?1 log10 VL drop but never undetectable on treatment) were randomized 2:1:1 to receive 48 weeks of peginterferon alfa-2a and ribavirin (PegIFN/RBV) in combination with faldaprevir 240 mg once daily (QD) with 3 days PegIFN/RBV lead-in (LI), 240 mg QD without LI, or 240 mg twice daily (BID) with LI. Patients in the 240 mg QD/LI group achieving maintained rapid virologic response (mRVR; VL &lt;25 IU/mL [Roche TaqMan] at week 4 and undetectable at weeks 8 to 20) were rerandomized to cease all treatment at week 24 or continue PegIFN/RBV up to week 48. Sustained virologic response (SVR) rates were 32%, 50%, and 42% in prior partial responders, and 21%, 35%, and 29% in prior null responders in the faldaprevir 240 mg QD/LI, 240 mg QD, and 240 mg BID/LI groups, respectively. In the 240 mg QD/LI group, a significantly higher proportion of mRVR patients rerandomized to 48 weeks' treatment achieved SVR compared with those assigned to 24 weeks treatment (72% versus 43%; P = 0.035). Rates of gastrointestinal disorders, jaundice, dry skin, and photosensitivity were increased at 240 mg BID compared with the 240 mg QD dose. Faldaprevir discontinuations owing to adverse events occurred in 6%, 4%, and 23% of patients in the 240 mg QD/LI, 240 mg QD, and 240 mg BID/LI groups, respectively. Conclusion: Faldaprevir 240 mg QD with PegIFN/RBV was safe and tolerable and produced substantial SVR rates in prior null and partial responders. The 240 mg QD dose is currently undergoing phase 3 evaluation. © 2013 American Association for the Study of Liver Diseases.
  • Author:
    Sulkowski MS; Asselah T; Lalezari J; Ferenci P; Fainboim H; Leggett B; Bessone F; Mauss S; Heo J; Datsenko Y; Stern JO; Kukolj G; Scherer J; Nehmiz G; Steinmann GG; Böcher WO
    Title:
    Faldaprevir combined with pegylated interferon alfa-2a and ribavirin in treatment-naïve patients with chronic genotype1 HCV: SILEN-C1 trial
    Source:
    Hepatology 57 (6), 2143-2154 (2013)
    Abstract:
    Faldaprevir (BI 201335) is a potent, hepatitis C virus (HCV) NS3/4A protease inhibitor with pharmacokinetic properties supportive of once-daily (QD) dosing. Four hundred and twenty-nine HCV genotype (GT)-1 treatment-naïve patients without cirrhosis were randomized 1:1:2:2 to receive 24 weeks of pegylated interferon alfa-2a and ribavirin (PegIFN/RBV) in combination with placebo, faldaprevir 120 mg QD with 3 days of PegIFN/RBV lead-in (LI), 240 mg QD with LI, or 240 mg QD without LI, followed by an additional 24 weeks of PegIFN/RBV. Patients in the 240 mg QD groups achieving maintained rapid virologic response (mRVR; viral load [VL] <25 IU/mL at week 4 and undetectable at weeks 8-20) were rerandomized to cease all treatment at week 24 or continue receiving PegIFN/RBV up to week 48. VL was measured by Roche TaqMan. Sustained virologic response (SVR) rates were 56%, 72%, 72%, and 84% in the placebo, faldaprevir 120 mg QD/LI, 240 mg QD/LI, and 240 mg QD groups. Ninety-two percent of mRVR patients treated with faldaprevir 240 mg QD achieved SVR, irrespective of PegIFN/RBV treatment duration. Eighty-two percent of GT-1a patients who received faldaprevir 240 mg QD achieved SVR versus 47% with placebo. Mild gastrointestinal disorders, jaundice resulting from isolated unconjugated hyperbilirubinemia, and rash or photosensitivity were more common in the active groups than with placebo. Discontinuations resulting from adverse events occurred in 4%, 11%, and 5% of patients treated with 120 mg QD/LI, 240 mg QD/LI, and 240 mg QD of faldaprevir versus 1% with placebo. Conclusion: Faldaprevir QD with PegIFN/RBV achieved consistently high SVR rates with acceptable tolerability and safety at all dose levels. The 120 and 240 mg QD doses are currently undergoing phase 3 evaluation. © 2013 American Association for the Study of Liver Diseases.
  • Author:
    Bailey MD; Bordeleau J; Garneau M; Leblanc M; Lemke CT; O'Meara J; White PW; Llinàs-Brunet M
    Title:
    Peptide backbone replacement of hepatitis C virus NS3 serine protease C-terminal cleavage product analogs: Discovery of potent succinamide inhibitors
    Source:
    Bioorg Med Chem Lett Article in press (2013)
    Abstract:
    A number of potent peptidic inhibitors of the NS3 protease have been described in the literature based on a substrate-based approach. In an on-going effort to reduce the peptidic character of this class of inhibitors, two novel series of analogs have been prepared in which the usual P3 amino acid residue is replaced by a succinamide fragment. This new backbone modification not only reduces the peptidic nature of traditional inhibitors but also provides new SAR opportunities for the capping group. Optimization of each of these two series resulted in inhibitors with sub-nanomolar potencies. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Bailey MD; Halmos T; Lemke C T
    Title:
    Discovery of novel P2 substituted 4-biaryl proline inhibitors of hepatitis C virus NS3 serine protease
    Source:
    Bioorg Med Chem Lett Article in Press (2013)
    Abstract:
    Inhibitors of hepatitis C virus NS3 serine protease often incorporate a large P2 moiety to interact with the surface of the enzyme while shielding part of the catalytic triad. This feature is important in many inhibitors in order to have the necessary potency needed for efficacy. In this Letter we explore some new P2 motifs to further exploit this region of the enzyme. In a continuing effort to replace the often found 4-hydroxyproline P2 core found in the majority of inhibitors for this target, various directly attached aryl derivatives were evaluated. Of these, the 2,4-disubstituted thiazole core proved to be the most interesting. SAR around this motif has lead to compounds with Ki's in the high picomolar range and provided cellular potencies in the single digit nM range. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Décor A; Grand-Maître C; Hucke O; O'Meara J; Kuhn C; Forget LC; Brochu C; Malenfant E; Bertrand-Laperle M; Bordeleau J; Ghiro E; Pesant M; Fazal G; Gorys V; Little M; Boucher C; Bordeleau S; Turcotte P; Guo T; Garneau M; Spickler C; Gauthier A
    Title:
    Design, synthesis and biological evaluation of novel aminothiazoles as antiviral compounds acting against human rhinovirus
    Source:
    Bioorg Med Chem Lett 23 (13), 3841-3847 (2013)
    Abstract:
    We describe here the design, synthesis and biological evaluation of antiviral compounds acting against human rhinovirus (HRV). A series of aminothiazoles demonstrated pan-activity against the HRV genotypes screened and productive structure-activity relationships. A comprehensive investigational library was designed and performed allowing the identification of potent compounds with lower molecular weight and improved ADME profile. 31d-1, 31d-2, 31f showed good exposures in CD-1 mice. The mechanism of action was discovered to be a host target: the lipid kinase phosphatidylinositol 4-kinase III beta (PI4KIIIß). The identification of the pan-HRV active compound 31f combined with a structurally distinct literature compound T-00127-HEV1 allowed the assessment of target related tolerability of inhibiting this kinase for a short period of time in order to prevent HRV replication. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Fader LD; Landry S; Morin S; Kawai SH; Bousquet Y; Hucke O; Goudreau N; Lemke CT; Bonneau P; Titolo S; Mason S; Simoneau B
    Title:
    Optimization of a 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of HIV capsid assembly inhibitors 1: Addressing configurational instability through scaffold modification
    Source:
    Bioorg Med Chem Lett Article in press (2013)
    Abstract:
    The optimization of a 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of inhibitors of HIV-1 capsid assembly that possess a labile stereocenter at C3 is described. Quaternization of the C3 position of compound 1 in order to prevent racemization gave compound 2, which was inactive in our capsid disassembly assay. A likely explanation for this finding was revealed by in silico analysis predicting a dramatic increase in energy of the bioactive conformation upon quaternization of the C3 position. Replacement of the C3 of the diazepine ring with a nitrogen atom to give the 1,5-dihydro-benzo[f][1,3,5]triazepine-2,4-dione analog 4 was well tolerated. Introduction of a rigid spirocyclic system at the C3 position gave configurationally stable 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione analog 5, which was able to access the bioactive conformation without a severe energetic penalty and inhibit capsid assembly. Preliminary structure-activity relationships (SAR) and X-ray crystallographic data show that knowledge from the 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of inhibitors of HIV-1 capsid assembly can be transferred to these new scaffolds. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Buckner C M; Moir S; Ho J; Wang W; Posada J G; Kardava L; Funk E K; Nelson A K; Li Y; Chun T-W; Fauci A S
    Title:
    Characterization of plasmablasts in the blood of HIV-infected viremic individuals: Evidence for nonspecific immune activation
    Source:
    J Virol 87 (10), 5800-5811 (2013)
    Abstract:
    Terminal differentiation of B cells and hypergammaglobulinemia are hallmarks of B-cell hyperactivity in HIV disease. Plasmablasts are terminally differentiating B cells that circulate transiently in the blood following infection or vaccination; however, in HIV infection, they arise early and are maintained at abnormally high levels in viremic individuals. Here we show that only a small fraction of plasmablasts in the blood of viremic individuals is HIV specific. Assessment of plasmablast immunoglobulin isotype distribution revealed increased IgG+ plasmablasts in early and most prominently during chronic HIV viremia, contrasting with a predominantly IgA+ plasmablast profile in HIV-negative individuals or in aviremic HIV-infected individuals on treatment. Of note, IgG is the predominant immunoglobulin isotype of plasmablasts that arise transiently in the blood following parenteral immunization. Serum immunoglobulin levels were also elevated in HIV-infected viremic individuals, especially IgG, and correlated with levels of IgG+ plasmablasts. Several soluble factors associated with immune activation were also increased in the sera of HIV-infected individuals, especially in viremic individuals, and correlated with serum immunoglobulin levels, particularly IgG. Thus, our data suggest that while plasmablasts in the blood may contribute to the HIV-specific immune response, the majority of these cells are not HIV specific and arise early, likely from indirect immune-activating effects of HIV replication, and reflect over time the effects of chronic antigenic stimulation. Such B-cell dysregulation may help explain why the antibody response is inadequate in HIV-infected individuals, even during early infection. © 2013, American Society for Microbiology.
  • Author:
    Fader L D; Landry S; Morin S; Kawai S H; Bousquet Y; Hucke O; Goudeau N; Lemke C T; Bonneau P; Titolo S; Mason S; Simoneau B
    Title:
    Optimization of a 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of HIV capsid assembly inhibitors 1: Addressing configurational instability through scaffold modification
    Source:
    Bioorg Med Chem Lett 23 (11), 3396-3400 (2013)
    Abstract:
    The optimization of a 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of inhibitors of HIV-1 capsid assembly that possess a labile stereocenter at C3 is described. Quaternization of the C3 position of compound 1 in order to prevent racemization gave compound 2, which was inactive in our capsid disassembly assay. A likely explanation for this finding was revealed by in silico analysis predicting a dramatic increase in energy of the bioactive conformation upon quaternization of the C3 position. Replacement of the C3 of the diazepine ring with a nitrogen atom to give the 1,5-dihydro-benzo[f][1,3,5]triazepine-2,4-dione analog 4 was well tolerated. Introduction of a rigid spirocyclic system at the C3 position gave configurationally stable 1,5-dihydrobenzo[b][1,4]diazepine-2,4- dione analog 5, which was able to access the bioactive conformation without a severe energetic penalty and inhibit capsid assembly. Preliminary structure-activity relationships (SAR) and X-ray crystallographic data show that knowledge from the 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of inhibitors of HIV-1 capsid assembly can be transferred to these new scaffolds. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Fader LD; Landry S; Goulet S; Morin S; Kawai S H; Bousquet Y; Dion I; Hucke O; Goudreau N; Lemke C T; Rancourt J; Bonneau P; Titolo S; Amad M; Garneau M; Duan J; Mason S; Simoneau B
    Title:
    Optimization of a 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of HIV capsid assembly inhibitors 2: Structure-activity relationships (SAR) of the C3-phenyl moiety
    Source:
    Bioorg Med Chem Lett 23 (11), 3401-3405 (2013)
    Abstract:
    Detailed structure-activity relationships of the C3-phenyl moiety that allow for the optimization of antiviral potency of a series of 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione inhibitors of HIV capsid (CA) assembly are described. Combination of favorable substitutions gave additive SAR and allowed for the identification of the most potent compound in the series, analog 27. Productive SAR also transferred to the benzotriazepine and spirobenzodiazepine scaffolds, providing a solution to the labile stereocenter at the C3 position. The molecular basis of how compound 27 inhibits mature CA assembly is rationalized using high-resolution structural information. Our understanding of how compound 27 may inhibit immature Gag assembly is also discussed. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Willkommen H; Blümel J; Brorson K; Chen D; Chen Q; Gröner A; Kreil T R; Robertson J S; Ruffing M; Ruiz S
    Title:
    Meeting report: PDA virus and TSE safety forum
    Source:
    PDA J Pharm Sci Technol 67 (2), 81-97 (2013)
    Abstract:
    The report provides a summary of the presentations and discussions of the Virus & TSE (transmissible spongiform encephalopathy) Safety Forum 2011 that was organized by the Parenteral Drug Association and held in Barcelona, Spain, on 28 -30 June, 2011. The conference was accompanied by a workshop named "Virus Removal by Filtration: Trends and New Developments." A summary of the workshop is provided as a separate report and will be published in this journal as well. The risk of virus contamination and mitigation strategies for medicinal products, sequence-based methods for virus detection, and virus reduction studies that characterize the capacity of specific unit operations for virus removal/ inactivation were reported during the Virus Safety Forum. The application of the design of experiment concept to virus safety studies, and the extensive work performed to understand the mechanism of action and to identify critical process parameters for virus removal/inactivation, have produced considerable data. They were provided during the conference and discussed. This report summarized not only the presented data; it also provides a summary of the panel discussion, which included representatives of regulatory agencies from different areas (USA, Europe, Japan) as well as experts from universities and industry. The TSE Safety Forum provided first an overview of the scientific data considering the occurrence of TSEs and the epidemiological situation in different areas. For production of cell-derived medicinal products, the risk of contamination occurs from bovine-derived raw materials like fetal bovine serum or from other raw materials produced with animal-derived components. The current risk of plasma-derived medicinal products from contamination of plasma with the variant Creutzfeldt-Jakob disease agent was considered, and gaps in knowledge and interpretation of TSE studies were discussed from the regulatory standpoint. Current understanding and gaps were intensively discussed by a panel of experts from universities, regulatory agencies and industries; they are summarized in this report. ©PDA, Inc. 2013.
  • Author:
    Willkommen H; Blümel J; Brorson K; Chen D; Chen Q; Gröner A; Kreil T R; Robertson J S; Ruffing M; Ruiz S
    Title:
    Meeting report - Workshop on virus removal by filtration: Trends and new developments
    Source:
    PDA J Pharm Sci Technol 67 (2), 98-104 (2013)
    Abstract:
    The workshop was held on 27 June 2011 in Barcelona, in conjunction with the PDA Virus & TSE (transmissible spongiform encephalopathy) Safety Forum 2011. Virus-retentive filters are important tools to assure a high virus safety level of biological medicinal products. Important parameters such as properties of virus spike preparations, mechanism of virus retention by different filter brands, use of prefilters to improve the filtration performance, and, finally, strategies to select the most appropriate filter for a specific product were discussed on the workshop. The panel discussion at the end of the workshop that involved speakers and regulators from different global areas came to following conclusions: o The major mechanism of virus retention is size exclusion; filtration, however, is complex and protein and virus can interact with the membrane in multiple ways. Pressure interruption during filtration resulted in enhanced virus passage. o It has never been reported that murine leukemia virus (MuLV) passes a parvovirus filter. It makes sense that a small virus can be used to provide a claim for a large virus like MuLV. This relies on the assumption that there is no aggregation or interaction of the model parvovirus with proteins leading to aggregates larger than retroviruses. o Several prefilters are under investigation to improve flow rate and throughput of filtration in large-scale manufacture. It was discussed whether the prefilter and the virus-retentive filter can be viewed as one unit operation so that virus retention by both can be claimed as the viral clearance capacity of this manufacturing step. This question engendered some controversy: whereas some saw the combination as a correct reflection of manufacturing conditions, others discussed the different mechanisms of virus retention, which need to be studied separately. All together, the workshop was seen as a valuable forum for the discussion between regulators and industry; it was proposed that such forum should be provided again if possible in connection with one of the next PDA Virus & TSE Safety Conferences. ©PDA, Inc. 2013
  • Author:
    Fader L D; Landry S; Goulet S; Morin S; Kawai S H; Bousquet Y; Dion I; Hucke O; Goudreau N; Lemke C T; Rancourt J; Bonneau P; Titole S; Amad M; Garneau M; Duan J; Mason S; Simoneau B
    Title:
    Optimization of a 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione series of HIV capsid assembly inhibitors 2: Structure-activity relationships (SAR) of the C3-phenyl moiety
    Source:
    Bioorg Med Chem Lett Article in press (2013)
    Abstract:
    Detailed structure-activity relationships of the C3-phenyl moiety that allow for the optimization of antiviral potency of a series of 1,5-dihydrobenzo[b][1,4]diazepine-2,4-dione inhibitors of HIV capsid (CA) assembly are described. Combination of favorable substitutions gave additive SAR and allowed for the identification of the most potent compound in the series, analog 27. Productive SAR also transferred to the benzotriazepine and spirobenzodiazepine scaffolds, providing a solution to the labile stereocenter at the C3 position. The molecular basis of how compound 27 inhibits mature CA assembly is rationalized using high-resolution structural information. Our understanding of how compound 27 may inhibit immature Gag assembly is also discussed. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Stammers T A; Coulombe R; Rancourt J; Thavonekham B; Fazal G; Goulet S; Jakalian A; Wernic D; Tsantrizos Y; Poupart M A; Bös M; McKercher G; Thauvette L; Kukolj G; Beaulieu P L
    Title:
    Discovery of a novel series of non-nucleoside thumb pocket 2 HCV NS5B polymerase inhibitors
    Source:
    Bioorg Med Chem Lett 23 (9), 2585-2589 (2013)
    Abstract:
    A novel series of non-nucleoside thumb pocket 2 HCV NS5B polymerase inhibitors were derived from a fragment-based approach using information from X-ray crystallographic analysis of NS5B-inhibitor complexes and iterative rounds of parallel synthesis. Structure-based drug design strategies led to the discovery of potent sub-micromolar inhibitors 11a-c and 12a-c from a weak-binding fragment-like structure 1 as a starting point. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    James C A; Deroy P; Duplessis M; Edwards P J; Halmos T; Minville J; Morency L; Morin S; Simoneau B; Tremblay M; Bethell R; Cordingley M; Duan J; Lamorte L; Pelletier A; rajotte D; Salois P; Tremblay S; Sturino C F
    Title:
    Nucleotide competing reverse transcriptase inhibitors: Discovery of a series of non-basic benzofurano[3,2-d]pyrimidin-2-one derived inhibitors
    Source:
    Bioorg Med Chem Lett 23 (9), 2781-2786 (2013)
    Abstract:
    A HTS screen led to the identification of a benzofurano[3,2-d]pyrimidin-2- one core structure which upon further optimization resulted in 1 as a potent HIV-1 nucleotide competing reverse transcriptase inhibitor (NcRTI). Investigation of the SAR at N-1 allowed significant improvements in potency and when combined with the incorporation of heterocycles at C-8 resulted in potent analogues not requiring a basic amine to achieve antiviral activity. Additional modifications at N-1 resulted in 33 which demonstrated excellent antiviral potency and improved physicochemical properties. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Stammers T A; Coulombe R; Rancourt J; Thavonekham B; Fazal G; Goulet S; Jakalian A; Wernic D; Tsantrizos Y; Poupart M A; McKercher G; Thauvett L; Kukolj G; Beaulieu P L
    Title:
    Discovery of a novel series of non-nucleoside thumb pocket 2 HCV NS5B polymerase inhibitors
    Source:
    Bioorg Med Chem Lett Article in Press (2013)
    Abstract:
    A novel series of non-nucleoside thumb pocket 2 HCV NS5B polymerase inhibitors were derived from a fragment-based approach using information from X-ray crystallographic analysis of NS5B-inhibitor complexes and iterative rounds of parallel synthesis. Structure-based drug design strategies led to the discovery of potent sub-micromolar inhibitors 11a-c and 12a-c from a weak-binding fragment-like structure 1 as a starting point. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Tremblay M; Bethell R C; Cordingley M G; DeRoy P; Duan J; Duplessis M; Edwards P J; Faucher A M; Halmos T; James C A; Kuhn C; Lacoste J E; Lamorte L; LaPlante S R; Malenfant E; Minville J; Moremcy L; Morin S; Rajotte D; Salois P; Simoneau B; Tremblay S; Sturino C F
    Title:
    Identification of benzofurano[3,2-d]pyrimidin-2-ones, a new series of HIV-1 nucleotide-competing reverse transcriptase inhibitors
    Source:
    Bioorg Med Chem Lett Article in Press (2013)
    Abstract:
    Screening of our sample collection led to the identification of a set of benzofurano[3,2-d]pyrimidine-2-one hits acting as nucleotide-competing HIV-1 reverse transcriptase inhibitiors (NcRTI). Significant improvement in antiviral potency was achieved when substituents were introduced at positions N1, C4, C7 and C8 on the benzofuranopyrimidone scaffold. The series was optimized from low micromolar enzymatic activity against HIV-1 RT and no antiviral activity to low nanomolar antiviral potency. Further profiling of inhibitor 30 showed promising overall in vitro properties and also demonstrated that its potency was maintained against viruses resistant to the other major classes of HIV-1 RT inhibitors. © 2013 Elsevier Ltd. All rights reserved.
  • Author:
    Harris L G; Nigam Y; Sawjer J; Mack D; Pritchard D I
    Title:
    Lucilia sericata chymotrypsin disrupts protein adhesin-mediated staphylococcal biofilm formation
    Source:
    Appl Environ Microbiol 79 (4), 1393-1395 (2013)
    Abstract:
    Staphylococcus aureus and Staphylococcus epidermidis biofilms cause chronic infections due to their ability to form biofilms. The excretions/secretions of Lucilia sericata larvae (maggots) have effective activity for debridement and disruption of bacterial biofilms. In this paper, we demonstrate how chymotrypsin derived from maggot excretions/secretions disrupts protein-dependent bacterial biofilm formation mechanisms. © 2013, American Society for Microbiology.
  • Author:
    Desforges M; Desjardins J; Zhang C; Talbot P J
    Title:
    The acetyl-esterase activity of the hemagglutinin-esterase protein of human coronavirus OC43 strongly enhances the production of infectious virus
    Source:
    J Virol 87 (6), 3097-3107 (2013)
    Abstract:
    Most betacoronaviruses possess an hemagglutinin-esterase (HE) protein, which appears to play a role in binding to or release from the target cell. Since this HE protein possesses an acetyl-esterase activity that removes acetyl groups from O-acetylated sialic acid, a role as a receptor-destroying enzyme has been postulated. However, the precise function of HE and of its enzymatic activity remains poorly understood. Making use of neutralizing antibody and of molecular clones of recombinant human coronavirus OC43 (HCoV-OC43), our results suggest that the HE protein of this HCoV could be associated with infection of target cells and, most notably, is important in the production of infectious viral particles. Indeed, after transfecting BHK-21 cells with various cDNA infectious clones of HCoV-OC43, either lacking the HE protein or bearing an HE protein with a nonfunctional acetyl-esterase enzymatic activity, we were reproducibly unable to detect recombinant infectious viruses compared to the reference infectious HCoV-OC43 clone pBAC-OC43FL. Complementation experiments, using BHK-21 cells expressing wild-type HE, either transiently or in a stable ectopic expression, demonstrate that this protein plays a very significant role in the production of infectious recombinant coronaviral particles that can subsequently more efficiently infect susceptible epithelial and neuronal cells. Even though the S protein is the main viral factor influencing coronavirus infection of susceptible cells, our results taken together indicate that a functionally active HE protein enhances the infectious properties of HCoV-OC43 and contributes to efficient virus dissemination in cell culture. © 2013, American Society for Microbiology.
  • Author:
    Wei X; Shu C; Haddad N; Zeng X; Patel N D; Tan Z; Liu J; Lee H; Shen S; Campbell S; Varsolona R J; Busacca C A; Hossain A; Yee N K; Senanayake C H
    Title:
    A highly convergent and efficient synthesis of a macrocyclic hepatitis C virus protease inhibitor BI 201302
    Source:
    Org Lett 15 (5), 1016-1019 (2013)
    Abstract:
    A highly convergent large scale synthesis of a 15-membered macrocyclic hepatitis C virus (HCV) protease inhibitor BI 201302 was achieved, in which the key features are the practical macrocyclization by Ru-catalyzed ring-closing metathesis (0.1 mol % Grela catalyst, 0.1-0.2 M concentration) and the efficient sulfone-mediated SNAr reaction. © 2013 American Chemical Society.
  • Author:
    Kurupati R; Tuyishime S; Kossenkov A V; Sazanovich M; Haut L H; Lasaro M O; Ratcliffe S J; Bosinger S E; Carnathan D G; Lewis M; Showe L C; Silvestri G; Ertl H C J
    Title:
    Correlates of relative resistance against low-dose rectal simian immunodeficiency virus challenges in peripheral blood mononuclear cells of vaccinated rhesus macaques
    Source:
    J Leukocyte Biol 93 (3), 437-448 (2013)
    Abstract:
    In this study, we compared the immunogenicity and protection from repeated low-dose intrarectal SIVmac251 challenge in two groups of vaccinated RMs. Animals were immunized with live SIVmac239, which had been attenuated by a deletion of the nef sequence, or they were vaccinated twice with an E1-deleted AdHu5, expressing SIVmac239gag. The vaccinated animals and a cohort of unvaccinated control animals were then challenged 10 times in weekly intervals with low doses of SIVmac251 given rectally. Our results confirm previous studies showing that whereas SIV?nef provides some degree of protection against viral acquisition after repeated low-dose rectal SIVmac251 challenges, vaccination with an AdHu5gag vaccine designed to induce only antiviral T cell responses is ineffective. As immunological analyses of prechallenge, vaccine-induced T and B cell responses failed to reveal correlates of protection that distinguished the more susceptible from the more resistant vaccinated animals, we carried out RNA-Seq studies of paired pre- and postvaccination samples to identify transcriptional patterns that correlated with the differences in response. We show that gene expression signatures associated with the delayed SIV infection seen in some AdHu5gag recipients were largely present in prevaccination samples of those animals. In contrast, the responding SIV?nef-immunized animals showed a predominance of vaccine-induced changes, thus enabling us to define inherited and vaccine-induced gene expression signatures and their associated pathways that may play a role in preventing SIV acquisition. © Society for Leukocyte Biology.
  • Author:
    Glover S; Anderson C; Piontkowski M; Ng T
    Title:
    Canine parvovirus (CPV) type 2b vaccine protects puppies with maternal antibodies to CPV when challenged with virulent CPV-2c virus
    Source:
    Int J appl res vet med 10 (3), 217-224 (2013)
    Abstract:
    Abstract This study was conducted to evaluate canine parvovirus disease prevention efficacy of the minimum immunizing dose of the CPV-2b fraction of a multivalent vaccine when administered at approximately 6 weeks of age to pups with maternal CPV-2b antibodies. A second dose was administered 4 weeks later. Pups were challenged with a virulent strain of CPV-2c virus 2 months after the second vaccination. Efficacy was evaluated by monitoring the pups for various clinical observations and laboratory testing of parvovirus infection, including mucous stool, bloody stool, diarrhea, fever, death, leukopenia, lymphopenia, CPV-2b serum neutralization titer, and detection of CPV in the feces. Upon a severe challenge with a virulent CPV-2c virus, four of five (80%) control pups had at least three of four clinical signs of CPV infection while 19 of 20 (95%) vaccinated pups had not more than one sign of CPV infection. The response of the control pups confirmed the virulence of the challenge and validity of the study. The response of the vaccinated pups demonstrated the efficacy of the CPV-2b vaccine, even in puppies with maternal antibody, which was one of the main objectives of this study. The outcome of this study was consistent with the 9CFR requirements necessary to support an additional label claim that the vaccine aids in the prevention of disease caused by CPV-2c when administered to puppies as young as 6 weeks of age with maternal CPV antibodies.
  • Author:
    O'Meara J A; Lemke C T; Goodbout C; Kukolj G; Lagacé L; Moreau B; Thibeault D; White P W; Linás-Brunet M
    Title:
    Molecular mechanism by which a potent hepatitis C virus NS3-NS4A protease inhibitor overcomes emergence of resistance
    Source:
    J Biol Chem 288 (8), 5673-5681 (2013)
    Abstract:
    Background: Antivirals must often be given in combinations to avoid rapid emergence of resistance. Results: We identified and structurally characterized protease inhibitors that maintain potency against genotype and resistant variants. Conclusion: Pan-variant potency was achieved by targeting invariant regions and incorporating flexibility where pocket variability occurs. Significance: Such inhibitors may yield simplified and/or more successful treatments for hepatitis C infections. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.
  • Author:
    Goudreau N; Columbe R; Faucher A M; Grand-Maitre C; Lacoste J E; Lemke C T; Malenfant E; Bousquet Y; Fader L; Simoneau B; Mercier J F; Titolo S; Mason S W
    Title:
    Monitoring Binding of HIV-1 Capsid Assembly Inhibitors Using 19F Ligand-and 15N Protein-Based NMR and X-ray Crystallography: Early Hit Validation of a Benzodiazepine Series
    Source:
    ChemMedChem 8 (3), 405-414 (2013)
    Abstract:
    The emergence of resistance to existing classes of antiretroviral drugs underlines the need to find novel human immunodeficiency virus (HIV)-1 targets for drug discovery. The viral capsid protein (CA) represents one such potential target. Recently, a series of benzodiazepine inhibitors was identified via high-throughput screening using an invitro capsid assembly assay (CAA). Here, we demonstrate how a combination of NMR and X-ray co-crystallography allowed for the rapid characterization of the early hits from this inhibitor series. Ligand-based 19FNMR was used to confirm inhibitor binding specificity and reversibility as well as to identify the N-terminal domain of the capsid (CANTD) as its molecular target. Protein-based NMR (1H and 15N chemical shift perturbation analysis) identified key residues within the CANTD involved in inhibitor binding, while X-ray co-crystallography confirmed the inhibitor binding site and its binding mode. Based on these results, two conformationally restricted cyclic inhibitors were designed to further validate the possible binding modes. These studies were crucial to early hit confirmation and subsequent lead optimization. © 2013 WILEY-VCH Verlag GmbH &amp; Co. KGaA, Weinheim.
  • Author:
    Huang Y-W; Harrall KK; Dryman BA; Opriessnig T; Vaughn EM; Roof MB; Meng X-J
    Title:
    Serological profile of torque teno sus virus species 1 (TTSuV1) in pigs and antigenic relationships between two tsuv1 genotypes (1a and 1b), between two species (TTSuV-1 and 2) and between porcine and human anelloviruses.
    Source:
    J Virol 86 (19), 10628-10639 (2012)
    Abstract:
    The family Anelloviridae includes human and animal torque teno viruses (TTVs) with extensive genetic diversity. The antigenic diversity among anelloviruses has never been assessed. Using torque teno sus virus (TTSuV) as a model, we describe here the first investigation of the antigenic relationships among different anelloviruses. Using a TTSuV genotype la (TTSuV1 a) or TTSuV1 b enzyme-linked immunosorbent assay (ELISA) based on the respective putative ORF1 capsid antigen and TTSuV1-specific real-time PCR, the combined serological and virological profile of TTSuV1 infection in pigs was determined and compared with that of TTSuV2. TTSuV1 is likely not associated with porcine circovirus-associated disease (PCVAD), because both the viral loads and antibody levels were not different between affected and unaffected pigs and because there was no synergistic effect of concurrent PCV2/TTSuV1 infections. We did observe a higher correlation of IgG antibody levels between anti-TTSuV1 a and -TTSuV1 b than between anti-TTSuV1 a or -1 b and anti-TTSuV2 antibodies in these sera, implying potential antigenic crossreactivity. To confirm this, rabbit antisera against the putative capsid proteins of TTSuV1 a, TTSuV1 b, or TTSuV2 were generated, and the antigenic relationships among these TTSuVs were analyzed by an ELISA and by an immunofluorescence assay (IFA) using PK-15 cells transfected with one of the three TTSuV ORF1 constructs. The results demonstrate antigenic crossreactivity between the two genotypes TTSuV1 a and TTSuV1 b but not between the two species TTSuV1 a or -1 b and TTSuV2. Furthermore, an antigenogroup 1 human TN antiserum did not react with any of the three TTSuV antigens. These results have important implications for an understanding of the diversity of anelloviruses as well as for the classification and vaccine development of TTSuVs. © 2012, American Society for Microbiology.
  • Author:
    Lemke CT; Titolo S; von Schwedler U; Goudreau N; Mercier J-F; Wardrop E; Faucher A-M; Coulombe R; Banik SSR; Fader L; Gagnon A; Kawai SH; Rancourt J; Tremblay M; Yoakim C; Simoneau B; Archambault J; Mason SW; Sundquist WI
    Title:
    Distinct effect of two HIV-1 capsid assembly inhibitor families that bind the same site within the N-termninal domain of the viral CA protein.
    Source:
    J Virol 86 (12), 6643-6655 (2012)
    Abstract:
    The emergence of resistance to existing classes of antiretroviral drugs necessitates finding new HIV-1 targets for drug discovery. The viral capsid (CA) protein represents one such potential new target. CA is sufficient to form mature HIV-1 capsids in vitro, and extensive structure-function and mutational analyses of CA have shown that the proper assembly, morphology, and stability of the mature capsid core are essential for the infectivity of HIV-1 virions. Here we describe the development of an in vitro capsid assembly assay based on the association of CA-NC subunits on immobilized oligonucleotides. This assay was used to screen a compound library, yielding several different families of compounds that inhibited capsid assembly. Optimization of two chemical series, termed the benzodiazepines (BD) and the benzimidazoles (BM), resulted in compounds with potent antiviral activity against wild-type and drug-resistant HIV-1. Nuclear magnetic resonance (NMR) spectroscopic and X-ray crystallographic analyses showed that both series of inhibitors bound to the N-terminal domain of CA. These inhibitors induce the formation of a pocket that overlaps with the binding site for the previously reported CAP inhibitors but is expanded significantly by these new, more potent CA inhibitors. Virus release and electron microscopic (EM) studies showed that the BD compounds prevented virion release, whereas the BM compounds inhibited the formation of the mature capsid. Passage of virus in the presence of the inhibitors selected for resistance mutations that mapped to highly conserved residues surrounding the inhibitor binding pocket, but also to the C-terminal domain of CA. The resistance mutations selected by the two series differed, consistent with differences in their interactions within the pocket, and most also impaired virus replicative capacity. Resistance mutations had two modes of action, either directly impacting inhibitor binding affinity or apparently increasing the overall stability of the viral capsid without affecting inhibitor binding. These studies demonstrate that CA is a viable antiviral target and demonstrate that inhibitors that bind within the same site on CA can have distinct binding modes and mechanisms of action.
  • Author:
    El-Far M; Szelei J; Yu O; Fediere G; Bergoin M; Tijssen P
    Title:
    Organization of the ambisense genome of the Helicoverpa armigera densovirus.
    Source:
    J Virol 86 (12), 7024 (2012)
    Abstract:
    A natural densovirus (DNV) of a serious phytophagous pest, Helicoverpa armigera, was isolated. The genome of HaDNV contained 6,039 nucleotides (nt) and included inverted terminal repeats (ITRs) of 545 nt with terminal Y-shaped hairpins of 126 nt. Its DNA sequence and ambisense organization with four typical open reading frames (ORFs) demonstrated that it belonged to the genus Densovirus in the subfamily Densovirinae of the family Parvoviridae
  • Author:
    Wang X-J; Zhang L; Sun X; Lee H; Krishnamurthy D; O'Meara JA; Landry S; Yoakim C; Simoneau B; Yee NK; Senanayake CH
    Title:
    Practical synthesis of a benzophenone-based NNRT inhibitor of HIV-1.
    Source:
    Org Process Res Dev 16 (4), 561-566 (2012)
    Abstract:
    A convergent synthesis of NNRTI 1 is described. The key step involves a direct coupling of acid chloride 4 with Grignard reagent 11 in the presence of bis[2-(N,N-dimethylamino)ethyl] ether that moderates the reactivity of the Grignard reagent to give benzophenone 7. An efficient 2-step process for the preparation of 2-fluoro-3-methyl-4-aminobenzoic acid (3) is also described. © 2012 American Chemical Society.
  • Author:
    Palmer S; Boltz VF; Chow JY; Martinson NA; McIntyre JA; Gray GE; Hopley MJ; Mayers D; Robinson P; Hall DB; Maldarelli F; Coffin JM; Mellors JW
    Title:
    Short-course Combivir after single-dose nevirapine reduces but does not eliminate the emergence of nevirapine resistance in women
    Source:
    Antiviral Ther 17 (2), 327-336 (2012)
    Abstract:
    Background: In the Treatment Options Preservation Study (TOPS) trial, 4 or 7 days of Combivir (CBV; zidovudine/lamivudine) with maternal single-dose nevirapine (sdNVP) significantly reduced the emergence of NVP resistance as determined by virus population genotyping. To detect NVP resistance with greater sensitivity, we analysed TOPS samples by allele-specific real-time PCR (ASP). Methods: In a random subset of women from each arm of the trial, plasma samples from before and 6 weeks after sdNVP were analysed using ASP at codons 103, 181, 184 and 190. Results: Samples were analysed from 27 women in the sdNVP arm and 24 each in the CBV 4-day (sdNVP/CBV4) and 7-day (sdNVP/CBV7) arms. ASP detected NVP-resistant variants in week 6 samples from 70% of women in the sdNVP arm, 29% in the sdNVP/CBV4 arm and 33% in sdNVP/CBV7 arm (P<0.01 for sdNVP/CBV4 or sdNVP/CBV7 versus sdNVP; P=1.0 for sdNVP/CBV4 versus sdNVP/CBV7). Lamivudine resistance was detected by ASP in only 1 of 51 women who received CBV. Conclusions: Short-course CBV significantly reduced but did not eliminate the emergence of NVP resistance after sdNVP. NVP-resistant variants were detected in about one-third of women despite CBV treatment, but the duration of persistence and clinical impact of these variants in response to antiretroviral therapy is uncertain. © 2012 International Medical Press.
  • Author:
    Marra A; Lamb L; Medina I; George D; Gibson G; Hardink J; rugg J; Van Deusen J; O'Donnell J P
    Title:
    Effect of linezolid on the 50% lethal dose and 50% protective dose in treatment of infections by gram-negative pathogens in naive and immunosuppressed mice and on the efficacy of ciprofloxacin in an acute murine model of septicemia
    Source:
    Antimicrob Agents Chemother 56 (9), 4671-4675 (2012)
    Abstract:
    Murine models of infection were used to study the effect of linezolid on the virulence of Gram-negative bacteria and to assess potential pharmacodynamic interactions with ciprofloxacin in the treatment of these infections, prompted by observations from a recent clinical trial. Naive and immunosuppressed mice were challenged with Klebsiella pneumoniae 53A1109, K. pneumoniae GC6658, and Pseudomonas aeruginosa UC12120 in acute sepsis and pulmonary infection models, using different serial dilutions of these pathogens (groups of 8 animals each). Linezolid (100 mg/kg/dose) was administered orally at 0.5 and 4.0 h postchallenge in the sepsis model and at 4 h postchallenge followed by 2 days of twice-daily treatment in the pulmonary model. Further, ciprofloxacin alone and in combination with oral linezolid was investigated in the sepsis model. Survival was assessed for 4 and 10 days postchallenge in the systemic and respiratory models, respectively. The data were fitted to a nonlinear regression analysis to determine 50% lethal doses (LD 50s) and 50% protective doses (PD 50s). A clinically relevant, high-dose regimen of linezolid had no significant effect on LD 50 in these models. This lack of effect was independent of immune status. A combination of oral ciprofloxacin with linezolid yielded lower PD 50s than oral ciprofloxacin alone (ciprofloxacin in combination, 8.4 to 32.7 mg/kg; oral ciprofloxacin, 39.4 to 88.3 mg/kg). Linezolid did not improve the efficacy of subcutaneous ciprofloxacin (ciprofloxacin in combination, 2.0 to 2.4 mg/kg; subcutaneous ciprofloxacin, 2.0 to 2.8 mg/kg). In conclusion, linezolid does not seem to potentiate infections caused by Gram-negative pathogens or to interact antagonistically with ciprofloxacin. Copyright © 2012, American Society for Microbiology. All Rights Reserved.
  • Author:
    Sun J-H; O'Boyle II DR; Zhang Y; Wang C; Nower P; Valera L; Roberts S; Nettles R E; Fridell R A; Gao M
    Title:
    Impact of a baseline polymorphism on the emergence of resistance to the hepatitis C virus nonstructural protein 5a replication complex inhibitor, BMS-790052.
    Source:
    Hepatology 55 (6), 1692-1699 (2012)
    Abstract:
    The influence of naturally occurring polymorphisms on the potency of the HCV nonstructural protein 5A (NS5A) replication complex inhibitor, BMS-790052, was investigated by evaluating hybrid replicons in which the entire NS5A coding region of genotype (GT) la and 1b laboratory (lab) strains (H77c and Con1) were replaced with the corresponding regions of specimens collected from 10 GT-1a- and 6 GT-1b-infected subjects. For baseline (BL) specimens, with no previously observed resistance variants identified by population sequencing, the median 50% effective concentration (EC 50) values for BMS-790052 were similar for the clinically derived and lab strains. A Q30R variant was observed at viral breakthrough (VBT) in one of the GT-1a-infected subjects. Because the lowest plasma exposure of BMS-790052 observed in this subject was 117 nM and the median 50% effective concentration value for a GT-1a H77c replicon containing a Q30R substitution is ?7 nM, a rigorous investigation was initiated to determine the basis for resistance. Three approaches were used: (1) replacement of the entire H77c NS5A or (2) replacement of the N-terminal region of NS5A, with sequence from BL and day 14, and (3) substitution of specific amino acids. A BL polymorphism (E62D) did not contribute resistance to BMS-790052; however, the linked variant, Q30R-E62D, conferred high-level resistance in vitro and is likely responsible for VBT in vivo. Conclusion: Our data show that a BL polymorphism with minimal effect on the anti-HCV effect of BMS-790052 can affect the emergence of resistance and significantly affect clinical outcome. This work establishes a clear, systematic approach to monitor resistance to NS5A inhibitors in the clinic. © 2012 American Association for the Study of Liver Diseases.
  • Author:
    Han Y-S; Quashie P; Mesplede T; Xu H; Mekhssian K; Fenwick C; Wainberg MA
    Title:
    A high-throughput assay for HIV-1 integrase 3'-processing activity using time-resolved fluorescence.
    Source:
    J Virol Methods 184 (1-2), 34-40 (2012)
    Abstract:
    HIV-1 integrase (HIV-1 IN), a well-validated antiviral drug target, catalyzes multistep reactions to incorporate viral DNA into the genome of the host cell; these include both a 3'-processing (3'P) reaction and a strand transfer reaction. These enzymatic activities can be measured in vitro with short DNA oligonucleotides that mimic a single viral LTR DNA end and purified IN. A highly sensitive and reproducible time-resolved fluorescence (TRF)-based assay for HIV-1 IN 3'P activity is now reported. This assay was optimized with respect to time and concentrations of metal ions, substrate and enzyme. The assay has now been used successfully to measure HIV-1 IN 3'P activity and has been shown to detect the anti-IN activity of several known 3'P inhibition compounds accurately. This assay, which is amenable to high-throughput screening, will be useful for identification of additional HIV-1 IN 3'P inhibitors. © 2012 Elsevier B.V.
  • Author:
    Ferris NP; Clavijo A; Yang M; Velazquez-Salinas L; Nordengrahn A; Hutchings GH; Kristersson T; Merza M
    Title:
    Development and laboratory evaluation of two lateral flow devices for the detection of vesicular stromatitis virus in clinical samples.
    Source:
    J Virol Methods 180 (1-2), 96-100 (2012)
  • Author:
    Arasteh K; Ward D; Plettenberg A; Livrozet JM; Orkin C; Cordes C; Guo J; Wang E; Yong CL; Robinson P; Quinson A
    Title:
    Twenty-four-week efficacy and safety of switching virologically suppressed HIV-1-infected patients from nevirapine immediate release 200mg twice daily to nevirapine extended release 400mg once daily (TRANxITION).
    Source:
    HIV Med 13 (4), 236-244 (2012)
  • Author:
    Larrey D; Lohse AW; de Ledinghen V; Trepo C; Gerlach T; Zarski JP; Tian A; Mathurin P; Thimme R; Arasteh K; Trautwein C; Cerny A; Dikopoulos N; Schuchmann M; Heim MH; Gerken G; Stern JO; Wu K; Abdallah N; Grilich B; Scherer J; Berger F; Marquis M; Kukkolj G; Boecher W; Steffgen J
    Title:
    Rapid and strong antiviral of the non-nucleosidic NSSB polymerase inhibitor BI207127 in combination with peginterfon alfa 2a and ribavirin.
    Source:
    J Hepatol Article in Press (2012)
  • Author:
    Ibrisimovic M; Nagl U; Kneidinger D; Rauch M; Lion T; Klein R
    Title:
    Targeted expression of herpes simplex virus thymidine kinase in adenovirus-infected cells reduces virus titers upon treatment with ganciclovir in vitro.
    Source:
    J Gene Med 14 (1), 3-19 (2012)
    Abstract:
    Background: Adenoviruses are a frequent cause of life-threatening infections in immunocompromised patients. Available therapeutics still cannot completely prevent fatal outcomes. By contrast, herpes viruses are well treatable with prodrugs such as ganciclovir (GCV), which are selectively activated in virus-infected cells by virus-encoded thymidine kinases. This effective group of prodrugs is not applicable to adenoviruses and other DNA viruses because they lack those kinases. Methods: To render adenoviruses amenable to GCV treatment, we generated an adenoviral vector-based delivery system for targeted expression of herpes simplex virus thymidine kinase (HSV-TK) in wild-type adenovirus 5 (wt Ad5)-infected cells. HSV-TK expression was largely restricted to wt virus-infected cells by transcription of the gene from the Ad5 E4 promoter. Its activity is dependent on the adenoviral E1A gene product which is not produced by the vector but is only provided in cells infected with wt adenovirus. The anti-adenoviral effect of HSV-TK expression and concomitant treatment with GCV was assessed in vitro in four different cell lines or primary cells. Results: E4 promoter-mediated HSV-TK background expression was sufficiently low to prevent cytotoxicity in the presence of low-levels GCV in cells not infected with wt Ad5. However, expression was several-fold increased in wt Ad5-infected cells and treatment with low levels of GCV efficiently inhibited wt Ad5 DNA replication. Genome copy numbers and output of infectious particles were reduced by up to>99.99% and cell viability was greatly increased. Conclusions: We extended the concept of enzyme/prodrug therapy to adenovirus infections by selectively sensitizing adenovirus-infected cells to treatment with GCV.
  • Author:
    Duan J; Garneau M; Amad Ma'An; Bolger G; De Marte J; Montpetit H; Otis F; Jutras M; Rheaume M; White PW; Bethell RC; Yong Ch-L; Llinas-Brunet M; Cordingley MG
    Title:
    Cross-species absorption, metabolism, distribution and pharmacokinetics of BI 201335, a potent HCV genotype 1 NS3/4A protease inhibtor.
    Source:
    Xenobiotica 42 (2), 164-172 (2012)
  • Author:
    Lavilla-Alonso S; Bauer MMT; Abo-Ramadan U; Ristimaeki A; Halavaara J; Desmond RA; Wang D; Escutenaire S; Ahtiainen L; Saksela K; Tatlisumak T; Hemminki A; Pesonen S
    Title:
    Macrophage metalloelastase (MME) as adjuvant for intra-tumoral injection of oncolytic adenovirus and its influence on metastases development.
    Source:
    Cancer Gene Ther 19 (2), 126-134 (2012)
    Abstract:
    Oncolytic adenoviruses are a promising treatment alternative for many advanced cancers, including colorectal cancer. However, clinical trials have demonstrated that single-agent therapy in advanced tumor masses is rarely curative. Poor spreading of the virus through tumor tissue is one of the major issues limiting efficacy. As oncolytic viruses kill preferentially cancer cells, high extracellular matrix (ECM) content constitutes potential barriers for viral penetration within tumors. In this study, the ECM-degrading proteases relaxin, hyaluronidase, elastase and macrophage metalloelastase (MME) were tested for their antitumor efficacy alone and in combination with oncolytic adenovirus. MME improved the overall antitumor efficacy of oncolytic adenovirus in subcutaneous HCT116 xenografts. In a liver metastatic colorectal cancer model, intra-tumoral treatment of primary tumors from HT29 cells with MME monotherapy or with oncolytic adenovirus inhibited tumor growth. Combination therapy showed no increased mortality in comparison with either monotherapy alone. Contradictory results of effects of MME on tumorigenesis and metastasis formation have been reported in the literature. This study demonstrates for the first time in a metastatic animal model that MME, as a monotherapy or in combination with oncolytic virus, does not increase tumor invasiveness. Co-administration of MME and oncolytic adenovirus may be a suitable approach for further optimization aiming at clinical applications for metastatic colorectal cancer.
  • Author:
    Schibler M; Gerlach D; Martinez Y; van Belle S; Kaiser L; Turin L; Tapparel C
    Title:
    Experimental human rhinovirus and enterovirus interspecies recombination.
    Source:
    J Gen Virol 93 (1), 93-101 (2012)
    Abstract:
    Human rhinoviruses (HRVs) and enteroviruses (HEVs), two important human pathogens, are nonenveloped, positive-sense RNA viruses of the genus Enterovirus within the family Picornaviridae. Intraspecies recombination is known as a driving force for enterovirus and, to a lesser extent, rhinovirus evolution. Interspecies recombination is much less frequent among circulating strains, and supporting evidence for such recombination is limited to ancestral events, as shown by recent phylogenetic analyses reporting ancient HRV-A/HRV-C, HEV-A/HEV-C and HEV-A/HEV-D recombination mainly at the 5'-untranslated region (5' UTR)-polyprotein junction. In this study, chimeric genomes were artificially generated using the 5' UTR from two different clinical HRV-C strains (HRV-Ca and HRV-Cc), an HRV-B strain (HRV-B37) and an HEV-A strain (HEV-A71), and the remaining part of the genome from an HRV-A strain (HRV-A16). Whilst the chimeric viruses were easily propagated in cell culture, the wild-type HRV-A16 retained a replication advantage, both individually and in competition experiments. Assessment of protein synthesis ability did not show a correlation between translation and replication efficiencies. These results reflect the interchangeability of the 5' UTR, including its functional RNA structural elements implicated in both genome translation and replication among different enterovirus species. The 5' UTR-polyprotein junction therefore represents a theoretic interspecies recombination breakpoint. This recombination potential is probably restricted by the need for co-infection opportunities and the requirement for the progeny chimera to outcompete the parental genomes' fitness, explaining the rare occurrence of such events in vivo.