Prior inhibition of AKT phosphorylation by BX795 can define a safer strategy to prevent herpes simplex virus-1 infection of the eye

Abstract

Purpose-To evaluate the prophylactic antiviral efficacy, corneal tolerance and toxicity of topically dosed BX795, a non-nucleoside small-molecule inhibitor of herpes simplex virus type-1 (HSV-1). Methods-Prophylactic treatment with BX795 was performed both in-vitro on human corneal epithelial cells and in-vivo on mice prior to HSV-1 challenge. Viral burden was evaluated using a standard plaque assay. In a separate experiment, mice were treated topically 3-times daily for 4weeks with BX795 to evaluate corneal tolerance and toxicity. Phenol-red thread measurements, fluorescein staining and optical coherence tomography (OCT) were used to evaluate tear production, dryness and corneal structural changes. Corneal sensitivity and intraocular pressure were measured using esthesiometery and tonometery respectively. Results-Both in-vitro and in-vivo results showed a robust suppression of HSV-1 infection when treated prophylactically with BX795. The fluorescein stain and phenol-red results for the BX795treated eyes did not show signs of corneal surface dryness when compared to trifluridine (TFT), an FDA-approved topical antiviral. The OCT measurements showed no signs of structural changes to the cornea suggesting that BX795 treatment was well tolerated without any apparent signs of toxicity or inflammation. The corneal sensitivity of BX795-treated eyes was not significantly different from TFT-treated eyes. No significant increase in the intraocular pressure of BX795treated mice was observed. Conclusions-Prophylactic treatment with BX795 protects corneal cells from HSV-1 infection. The antiviral is well-tolerated on murine corneas without any detectable toxicity.

Hsp90-associated DNA replication checkpoint protein and proteasome-subunit components are involved in the age-related macular degeneration

“…[ 26 ] BX795 is a multiple kinase inhibitor which suppresses the inflammatory response and is safe for eye treatment. [ 29 , 30 ] Bortezomib and a few other drugs have shown potential anti-tumor activities. [ 31 ]…”

Standalone or combinatorial phenylbutyrate therapy shows excellent antiviral activity and mimics CREB3 silencing

“…Cell viability assay using various concentrations of PBA/NaPBA was performed on HCE cells plated at a density of 1 × 10 4 per well in 96-well plates overnight ( 45 ). Concentrations starting at 100 mM were twofold serially diluted in whole media and added to cell monolayers for a period of 24 hours.…”

“…At 28 dpi, the mice were euthanized, followed by cervical dissociation, and their trigeminal ganglia were incubated in DMEM whole media. Four days after incubation, the supernatants from the trigeminal ganglia suspension were titered for viral reactivation from using a plaque assay on Vero cells ( 45 ).…”

“…All the ocular toxicity studies including surface dryness, optical coherence tomography, corneal sensitivity, and ocular hypertension were performed similar to methods mentioned previously by our laboratory ( 45 ). Drugs were administered three times every day for 4 weeks, and the readings were procured every 2 weeks.…”

The 3- O -sulfation of heparan sulfate modulates protein binding and lyase degradation

“…A fluorescence assay using GFP reporter HSV-1 virions mixed with the hexasaccharides (1A to 1C through 9A to 9C, at 100 μg/mL concentration) was performed to determine the ratio of infected cells vs. total cells (DAPI nuclear stain) (Fig. 6A) (48,49). Next, IC 50 was determined for compounds exhibiting potent inhibition at 100 μg/mL concentration (7A, 7C, 8A, 8C, 9A, and 9C; Fig.…”