Prodrugs of antiviral phosphonates

Phosphonoformate and phosphonoacetate are effective antiviral agents, however they are charged at physiological pH and as such penetration into cells and diffusion across the blood-brain bamer is limited. In an attempt to increase the lipophilicity and improve the transport properties of these molecules, prodrugs were synthesised and their stabilities and reconversion to the parent compound subsequently investigated by the techniques of 31P nuclear magnetic resonance spectroscopy and high performance liquid Chromatography. A series of 4-substituted dibenzyl (methoxycarbonyl)phosphonates were prepared and found to be hydrolytically unstable giving predominantly the diesters, benzyl (methoxycarbonyl)phosphonates. This instability arose from the electron-withdrawing effect of the carbonyl group promoting nucleophilic attack at phosphorus. It was possible to influence the mechanism and, to some extent, the rate of hydrolysis of the phosphonoformate triesters to the diesters by varying the electronic nature of the substituent in the 4-position of the aromatic ring. Strongly electron-withdrawing groups increased the sensitivity of phosphorus to nucleophilic attack, thus promoting P-O .bond cleavage and rapid hydrolysis. Conversely, weakly electron-withdrawing substituents encouraged C-O bond fission, presumably through resonance stabilisation of the benzyl carbonium ion. The loss of the protecting group on phosphorus was in competition with nucleophilic attack at the carbonyl group, resulting in P-C bond cleavage with dibenzyl phosphite formation. The high instability and P-C bond fission make triesters unsuitable prodrug forms of phosphonoformate. A range of chemically stable triesters of phosphonoacetate were synthesised and their bioactivation investigated. Di(benzoyloxymethyl) (methoxycarbonylmethyl)phosphonates degraded to the relevant benzoyloxymethyl (methoxycarbonylmethyl)phosphonate in the presence of esterase. The enzymatic activation was restricted to the removal of only one protecting group from phosphorus, most likely due to the close proximity of the benzoyloxy ester function to the anionic charge on the diester. However, in similar systems di(4-alkanoyloxybenzyl) (methoxycarbonylmethyl)phosphonates degraded in the presence of esterase with the loss of both protecting groups on phosphorus to give the monoester, (methoxycarbonylmethyl)phosphonate, via the intermediary of the unstable 4-hydroxy benzyl esters. The methoxycarbonyl function remained intact. The rate of enzymatic hydrolysis and subsequent removal of the protecting groups on phosphorus was dependent on the nature of the alkanoyl group and was most rapid for the 4-nbutanoyloxybenzyl and 4-iso-butanoyloxybenzyl esters of phosphonoacetate. This provides a strategy for the design of a prodrug with sufficient stability in plasma to reach the central nervous system in high concentration, wherein rapid metabolism to the active drug by brain-associated enzymes occurs.

The evolutionary history and activity of a gain-of-function polymorphism in a human antiviral gene

Thesis (Ph.D.)--University of Washington, 2016-07

Antiviral host defence peptides.

The on going global mortality and morbidity associated with viral pathogens highlights the need for the continued development of effective, novel antiviral molecules. The antiviral activity of cationic host defence peptides is of significant interest as novel therapeutics for treating viral infection and predominantly due to their broad spectrum antiviral activity. These peptides also display powerful immunomodulatory activity and are key mediators of inflammation. Therefore, they offer a significant opportunity to inform the development of novel therapeutics for treating viral infections by either directly targeting the pathogen or by enhancing the innate immune response. In this chapter, we review the antiviral activity of cathelicidins and defensins, and examine the potential for these peptides to be used as novel antiviral agents.

Actividad bactericida, antifúngica y antiviral de nanopartículas de bismuto contra patógenos orales

De acuerdo con un anuncio publicado por el NIH (National Institute of Health, EUA), los biofilms o biocapas son un problema de salud pública muy importante en el área médica, ya que representan más del 80% de las infecciones microbianas en el cuerpo humano. Los tratamientos antimicrobianos estándar, típicamente fallan para erradicarlos. Por tanto, existe una creciente necesidad de desarrollo de nuevos fármacos con nuevas y mejores capacidades que puedan controlar a los biofilms. Hoy en día, la nanoterapeútica ofrece opciones innovadoras para controlar enfermedades infecciosas incluyendo la cavidad oral, contrarrestando el crecimiento de microorganismos patógenos tales como, Streptococcus mutans, Candida albicans y herpesvirus. En el área médica, algunos derivados del bismuto, como el subsalicilato, han sido empleados en el área médica para contrarrestar el vómito, náuseas, diarrea y dolor de estómago. En esta investigación determinamos la actividad antimicrobiana de nanopartículas de bismuto (BiNPs) contra patógenos orales. La actividad antibacteriana y antifúngica lo evaluamos por el ensayo de MTT y microscopía de fluorescencia. Demostramos que las BiNPs inhibieron el crecimiento de Streptococcus mutans con un 69% de efectividad así mismo obtuvimos un 85% de inhibición con Candida albicans, además fueron capaces de inhibir la formación del biofilm de ambas cepas. Por otra parte, determinamos la actividad antiviral en un modelo de rotavirus mediante el ensayo de inmunoperoxidasa presentando 90% de control de infección con el tratamiento de BiNPs. Finalmente, se evaluó la posible citotoxicidad de BiNPs en células epiteliales de riñón de mono mediante microscopía de fluorescencia y no mostró evidencias de citotoxicidad a 24 horas de exposición.

In vitro characterization of the antiviral activity of fucoidan from Cladosiphon okamuranus against Newcastle Disease Virus

Newcastle Disease Virus (NDV) causes a serious infectious disease in birds that results in severe losses in the worldwide poultry industry. Despite vaccination, NDV outbreaks have increased the necessity of alternative prevention and control measures. Several recent studies focused on antiviral compounds obtained from natural resources. Many extracts from marine organisms have been isolated and tested for pharmacological purposes, and their antiviral activity has been demonstrated in vitro and in vivo. Fucoidan is a sulfated polysaccharide present in the cell wall matrix of brown algae that has been demonstrated to inhibit certain enveloped viruses with low toxicity. This study evaluated the potential antiviral activity and the mechanism of action of fucoidan from Cladosiphon okamuranus against NDV in the Vero cell line

Determination of antioxidant activity, phenolic contents and antiviral potential of methanol extract of Euphorbia spinidens (Euphorbiaceae)

Purpose: This study was aimed to evaluate the antioxidant activity of the methanol extract of Euphorbia spinidens (Euphorbiaceae) and its effect on Herpes simplex virus type-1 (HSV-1) replication. Methods: The methanol extract of aerial parts of E. spinidens collected from Khorasan State in North- Eastern part of Iran was used in this study. Total phenolic, flavonoid contents and the antioxidant activity were evaluated using Folin-Ciocalteu method, aluminum chloride colorimetric method and β- carotene-linoleate model system, respectively. Both the cytotoxic and antiviral effects of the crude extract on Vero cell line were determined by quantifying the viability of Vero cells using 3-(4,5- dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium (MTS) assay. Results: Total phenolic and flavonoid contents of E.spinidens were 70 ± 1 mg of gallic acid equivalent/g of dry extract (mg GAE/g extract) and 49.66 ± 1.00 mg rutin equivalent/g of dry extract (mg RTN/g extract), respectively. Antioxidant activity was 44 ± 1 % compared with the standard, buthylated hydroxytuloene (BHT). The 50 % cytotoxic concentration (CC50) of the extract on Vero cells was 5.072 ± 0.063 mg/ml and its antiviral concentration of 50 % effectiveness (EC50) value was 0.34 ± 0.003 mg/ml. Conclusion: The findings of this study show that the methanol extract of E. spinidens has high content of phenolic and flavonoid compounds with good antioxidant activity. Furthermore, this extract has significant antiviral effect on HSV-1 probably due to the inhibition of viral replication.

Antiviral activity and mechanism of action of arbidol against Hantaan virus infection

Purpose: To investigate the activity and mechanism of action of arbidol against Hantaan virus (HTNV) activity by modulating inflammation via TLR-4 pathway. Methods: HUVEC cells infected with HTNV 76-118 were treated with serially diluted arbidol solutions at -2h (2 h before viral infection, pre-treatment mode), 0 h (at the same time as viral infection, simultaneous treatment mode) or 2 h (2 h after viral infection, post-treatment mode). The transcript levels of TLR4 were detected by semi-quantitative reverse transcription-PCR (RT-PCR) at 6, 12, 18, and 24 h later. The levels of iNOS and TNF-α were examined using enzyme-linked immunosorbent assay (ELISA). Results: Pre-treatment with arbidol, rather than simultaneous treatment or post-treatment, effectively inhibited up-regulation of cellular TLR4 expression (up to 40 ± 6.1 % inhibition) and activity of supernatant iNOS induced by HTNV infection(up to 44.1 ± 9.4 % inhibition), as well as in a LPSstimulated inflammatory endothelial cell. Arbidol decreased the elevated TNF-α levels induced by LPS stimulation. Conclusion: These results are the first evidence that arbidol modulates viral PRRs signaling and its consequential inflammatory cytokine/chemokine response during hantavirus infection.

Selective accumulation of virus-specific CD8+ T cells within the peripheral blood stem cell compartment

The absence of cellular immunity is central to the pathogenesis of herpesvirus-mediated diseases after allogeneic hemopoietic stem cell transplantation (HSCT). For both bone marrow (BM)– and granulocyte-colony stimulating factor–mobilized peripheral blood stem cells (PBSCs) HSCT, donor-derived Epstein-Barr virus (EBV) and cytomegalovirus (CMV) peptide–specific CD8+ T cells clones undergo early expansion and persist long-term, with additional diversification arising from novel antigen-specific clones from donor-derived progenitors. Whether BM or PBSC is the superior source of antiviral CD8+ T cells is unclear. Given that PBSC has largely replaced BM as a source of stem cells for HSCT, it is unlikely that herpesvirus effector T-cell reconstitution will ever be compared prospectively. PBSC grafts contain 10 to 30 times more T cells than BM and a randomized study found proven viral infections were more frequent in BM than PBSC recipients, suggesting viral-specific T-cell immunity is enhanced in PBSC. Recently Moss showed in lung cancer patients that herpesvirus-specific BM-derived CD8+ T cells have unique homing properties relative to herpesvirus-specific CD8+ T cells present in unmobilized peripheral blood (PB). Immunodominant EBV-lytic peptide–specific CD8+ T cells were enriched in BM but were reduced for CMV peptide–specific CD8+ T cells relative to PB. EBV-latent peptide–specific CD8+ T cells were equivalent, which has relevance in the context of posttransplantation lymphoproliferative disorder for which impaired EBV-latent CD8+ T-cell immunity is a risk-factor. A comparison of herpesvirus-specific cellular immunity in PBSC versus PB has yet to be performed.