ABSTRACT
AIMS: To investigate the anti-HSV and anti-inflammatory effects of a standardized ethyl acetate extract (SEAE) prepared with the stem bark of Strychnos pseudoquina, along with two isolated compounds: quercetin 3-O-methyl ether (3MQ) and strychnobiflavone (SBF). METHODS AND RESULTS: The mechanisms of action were evaluated by different methodological strategies. SEAE and SBF affected the early stages of viral infection and reduced HSV-1 protein expression. Both flavonoids elicited a concentration-dependent inhibition of monocyte chemoattractant protein-1 (MCP-1), whereas 3MQ reduced the chemokine release more significantly than SBF. Conversely, both compounds stimulated the production of the cytokines TNF-α and IL-1-ß in LPS-stimulated cells, especially at the intermediate and the highest tested concentrations. CONCLUSIONS: SEAE and SBF interfered with various steps of HSV replication cycle, mainly adsorption, postadsorption and penetration, as well as with ß and γ viral proteins expression; moreover, a direct inactivation of viral particles was observed. Besides, both flavonoids inhibited MCP-1 selectively, a feature that may be beneficial for the development of new anti-HSV agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicated that the samples present anti-HSV and anti-inflammatory activities, at different levels, which is an interesting feature since cold and genital sores are accompanied by an inflammation process.
Subject(s)
Antiviral Agents/pharmacology , Biflavonoids/pharmacology , Herpesvirus 1, Human/drug effects , Quercetin/analogs & derivatives , Strychnos/chemistry , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Antiviral Agents/chemistry , Biflavonoids/chemistry , Brazil , Cell Line , Chemokine CCL2/metabolism , Chlorocebus aethiops , Cytokines/metabolism , Herpesvirus 1, Human/physiology , Humans , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plants, Medicinal/chemistry , Quercetin/chemistry , Quercetin/pharmacology , Vero CellsABSTRACT
In this report, we described the genistein distribution on excised porcine esophageal mucosa from cationic nanoemulsions, as well as the anti-HSV-1 activity against a viral strain resistant to acyclovir. Genistein-loaded cationic nanoemulsions were prepared by spontaneous emulsification. This procedure yielded monodisperse nanoemulsions exhibiting a mean droplet size of approximately 200-300 nm. Hydroxyethyl cellulose (HEC) was added at the end of the manufacturing process as a thickening agent (at 3%). Such formulations exhibit a non-Newtonian pseudoplastic behavior. The addition of HEC significantly reduces the genistein flux through excised porcine mucosa specimens as compared with values elicited by nanoemulsions before thickening. Furthermore, a significant increase of genistein retention in mucosa was observed as compared to the genistein propylene glycol solution, as illustrated by confocal fluorescence microscopy images. Formulations exhibited antiherpetic activity in vitro against HSV-1 (strain 29R). Taken together, these results suggest that these formulations have promising potential to be used topically for herpes infections.
Subject(s)
Antiviral Agents , Drug Carriers , Genistein , Mucous Membrane/metabolism , Nanoparticles/chemistry , Animals , Antiviral Agents/chemistry , Antiviral Agents/pharmacokinetics , Antiviral Agents/pharmacology , Chlorocebus aethiops , Drug Carriers/chemistry , Drug Carriers/pharmacokinetics , Drug Carriers/pharmacology , Drug Evaluation, Preclinical , Emulsions , Genistein/chemistry , Genistein/pharmacokinetics , Genistein/pharmacology , Permeability , Swine , Vero CellsABSTRACT
The antiviral effects of soybean isoflavonoids have been investigated recently, especially those of genistein. It has been reported that this isoflavone is able to inhibit herpes simplex virus (HSV) replication, which is associated with skin and epithelial mucosa infections. The treatment of these infections with antiherpes drugs has resulted in the emergence of resistant viral strains. Based on this evidence, the aim of this study was to investigate the anti-HSV effects of soybean isoflavonoids: daidzein, genistein, glycitein, and coumestrol. Genistein and coumestrol inhibited HSV-1 (KOS and 29R strains, which are acyclovir sensitive and acyclovir resistant, respectively) and HSV-2 (333 strain) replication, whereas no antiviral effects were detected for daidzein and glycitein. The mechanisms of action were evaluated by different methodological strategies. Coumestrol affected the early stages of viral infection, and both compounds were able to reduce HSV-1 protein expression, as well as HSV-2 cell-to-cell spread.
Subject(s)
Glycine max/chemistry , Herpesvirus 1, Human/drug effects , Herpesvirus 2, Human/drug effects , Isoflavones/pharmacology , Virus Replication/drug effects , Antiviral Agents/isolation & purification , Antiviral Agents/pharmacology , Herpesvirus 1, Human/physiology , Herpesvirus 2, Human/physiology , Humans , Isoflavones/isolation & purificationABSTRACT
A simple, rapid, and sensitive LC method to determine coumestrol incorporated in the lipid nanoemulsions was validated. The analyses were performed at room temperature on a reversed-phase C18 column using a mobile phase composed of methanol/water with 0.1% trifluoracetic acid (70:30, v/v) at 0.8 mL min(-1). The detection was carried out on a UV detector at 343 nm. The linearity, in the range of 0.1-6.0 microg/mL, presented a determination coefficient (r2) of 0.999, calculated by the least square method. No interferences of the oil core or the gelling excipients were detected. The R.S.D. values for intra- and inter-day precision experiments were lower than 2%. The recovery ranged from 99.42% to 100.72%. Finally, the proposed method was successfully applied to determine coumestrol incorporated in the proposed topical formulations.
