Effect of vitamin D3 supplementation on the pharmacokinetics of digoxin : a pilot study

Emerging evidence from preclinical, clinical and epidemiological studies suggests that vitamin D3 plays vital roles in several diseases in addition to bone disorders. According to new medical evidence, it is being recommended that vitamin D3 intake to be increased for maximal benefits in human health. However, it is necessary to consider potential side effects of increased intake of vitamin D3. Vitamin D3 exerts its actions through the vitamin D receptor, which is known to be an important regulator of P-glycoprotein (P-gp). As P-gp plays a significant role in limiting drug bioavailability, we undertook a study to compare single-dose digoxin (a P-gp substrate) pharmacokinetics in eight healthy male subjects before and after vitamin D3 supplementation (1000IU per day). The geometric mean ratios for AUC0-3h, AUC0-48h and Cmax were 1.06 (90% CI 0.92, 1.21) and 1.02 (90% CI 0.97, 1.08) and 1.03 (95% CI 0.86, 1.24), respectively. The median for digoxin Tmax was 0.75h before and after vitamin D3 ingestion. The mean plasma 25-hydroxyvitamin D3 (25(OH)D3) levels remained constant after the intake of vitamin D3 (15.4±3.7 and 14.4±3.6ng/mL, respectively), while there was a modest but statistically significant increase in plasma calcium levels, from 9.32-9.68mg/dL (P=0.0277). These results suggest that vitamin D3 supplementation (1000IU per day) in human volunteers does not produce a P-gp-mediated drug interaction with orally administered digoxin

Generation of Combinatorial Lentiviral Vectors Expressing Multiple Anti-Hepatitis C Virus shRNAs and Their Validation on a Novel HCV Replicon Double Reporter Cell Line

Despite the introduction of directly acting antivirals (DAAs), for the treatment of hepatitis C virus (HCV) infection, their cost, patient compliance, and viral resistance are still important issues to be considered. Here, we describe the generation of a novel JFH1-based HCV subgenomic replicon double reporter cell line suitable for testing different antiviral drugs and therapeutic interventions. This cells line allowed a rapid and accurate quantification of cell growth/viability and HCV RNA replication, thus discriminating specific from unspecific antiviral effects caused by DAAs or cytotoxic compounds, respectively. By correlating cell number and virus replication, we could confirm the inhibitory effect on the latter of cell over confluency and characterize an array of lentiviral vectors expressing single, double, or triple cassettes containing different combinations of short hairpin (sh)RNAs, targeting both highly conserved viral genome sequences and cellular factors crucial for HCV replication. While all vectors were effective in reducing HCV replication, the ones targeting viral sequences displayed a stronger antiviral effect, without significant cytopathic effects. Such combinatorial platforms as well as the developed double reporter cell line might find application both in setting-up anti-HCV gene therapy approaches and in studies aimed at further dissecting the viral biology/pathogenesis of infection