furaprofen

R803 is an effective HCV replication inhibitor. R803 is substantially more potent against genotype 1a and 1b replicons (EC50, ~30 nM) than against the genotype 2a replicon (EC50, ~1,000 nM).

Signaling Pathways >> Anti-infection >> HCV

Research Areas >> Infection

EC50: ~30 nM (HCV genotype 1a and 1b replicons), ~1000 nM (HCV genotype 2a replicon)[1]

R803 is potent and highly specific for HCV replication. The antiviral activity of R803 has been determined by a reporter replicon assay with multiple repeats to be 29.88±8.05 nM, an ~3-fold improvement over the activity of the parent compound, R706. The potency of R803 against the replicon is also confirmed by both Western blotting and TaqMan RT-PCR to be about 37 nM and 54.67±4.11 nM, respectively . To assess the general effect of R803 on cell proliferation, a panel of primary cells and transformed human cell lines are treated with increasing doses of R803 for 48 h, and the effect on cell proliferation is measured by an MTS-based cell viability assay. The the concentration that caused a 50% reduction in cell numbers in the absence of virus infection (CC50) of R803 is found to range from 2 μM to ≥10 μM, depending on the cell type and proliferation status[1].

Plasmid PV-Luc is linearized with the PvuI restriction enzyme and subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7. After DNase I digestion and purification, the transcribed RNA (5 μg) is electroporated into Huh-7 cells at 270 V and 950 μF on a Gene Pulser II system. The cells are then dispensed into white 96-well plates at a density of 10,000/90 μL/well. R803 is serially diluted in compound diluent (10% DMSO, 20% methanol) and added to each well in a volume of 10 μL. The plates are incubated for 18 h at 37°C under 5% CO2 before luciferase activity is determined using the Bright-Glo luciferase assay kit[1].

Replicon 9-13 cells are plated onto 6-well plates 24 h prior to the treatment. Serial dilutions of R803 are made in a mixture containing 90% of the culture medium, 7.2% 1× PBS, 1.8% methanol, 1% DMSO, 20 μM RBV, and varying concentrations of IFN-α for a fixed-ratio dose-response study. The cells are treated with the designated combinations of R803 (0 to 80 nM concentrations) and IFN-α (0 to 4 IU/mL) plus 20 μM RBV for 72 h; then they are washed with PBS, lysed in SDS loading buffer, and analyzed by Western blotting[1].

[1]. Huang P, et al. Discovery and characterization of substituted diphenyl heterocyclic compounds as potent and selective inhibitors of hepatitis C virus replication. Antimicrob Agents Chemother. 2008 Apr;52(4):1419-29.

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MOLECULAR FORMULA :

C17-H14-O3

MOLECULAR WEIGHT :

266.31

WISWESSER LINE NOTATION :

T56 BOJ DR& IY1&VQ

HEALTH HAZARD DATA

ACUTE TOXICITY DATA

TYPE OF TEST :

TDLo - Lowest published toxic dose

ROUTE OF EXPOSURE :

Oral

SPECIES OBSERVED :

Human

DOSE/DURATION :

5 mg/kg

TOXIC EFFECTS :

Gastrointestinal - other changes

REFERENCE :

JCPCBR Journal of Clinical Pharmacology. (Hall Assoc., PO Box 482, Stamford, CT 06904) V.13- 1973- Volume(issue)/page/year: 16,473,1976

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Oral

SPECIES OBSERVED :

Rodent - rat

DOSE/DURATION :

950 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

REFERENCE :

AIPTAK Archives Internationales de Pharmacodynamie et de Therapie. (Heymans Institute of Pharmacology, De Pintelaan 185, B-9000 Ghent, Belgium) V.4- 1898- Volume(issue)/page/year: 214,240,1975

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Oral

SPECIES OBSERVED :

Mammal - dog

DOSE/DURATION :

>1400 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

REFERENCE :

AIPTAK Archives Internationales de Pharmacodynamie et de Therapie. (Heymans Institute of Pharmacology, De Pintelaan 185, B-9000 Ghent, Belgium) V.4- 1898- Volume(issue)/page/year: 214,240,1975