Nature research paper: Blocking NS3–NS4B interaction inhibits dengue virus in non-human primates
. DENV-1 RNA was detected in a 25 µl reaction containing 5 µl of extracted RNA sample, 10 pmol of dengue-specific primers and 5 pmol dengue probe in the reaction mixture from Superscript Ill Platinum One-Step Quantitative RT-PCR Kit , consisting of 0.5 µl Superscript III RT/Platinum mix, 12.5 µl 2× reaction mix and 0.05 µl ROX reference dye.
The pharmacokinetics profile of JNJ-1802 was evaluated in female monkeys after single intravenous administration and in DENV-infected monkeys after repeated oral administration. After single intravenous administration, the monkey blood was collected at the BPRC just before dosing, and at 0.05, 0.15, 0.5, 4, 6, 8, 24, 48 and 264 h after dosing.
To assess the effect of JNJ-1802 treatment on viral load in DENV-2-infected mice for each treatment group compared with the vehicle-treated mice , ordinary one-way ANOVA was used with Dunnett’s test to correct for multiple comparisons and the pooled data of three independent studies were analysed using two-way ANOVA with Dunnett’s test to correct for multiple comparisons .
To assess the effect of JNJ-1802 treatment on viral load in DENV-1-, DENV-3- or DENV-4-infected mice for each treatment group compared with the vehicle-treated mice , ordinary one-way ANOVA was applied with Dunnett’s test to correct for multiple comparisons , and Kruskal–Wallis tests with Dunn’s multiple-comparison test . For the viral kinetics studies and the delayed-treatment studies, a batch approach was applied to calculate the viral load AUC using the PΚ R package.
To assess the antiviral activity of JNJ-1802 in rhesus macaques infected with the DENV-1/45AZ5 virus strain, the sample size was calculated using one-sided Fisher exact tests and an= 0.05 targeting a power of at least 80%. This sample size was sufficient to detect a significant difference in the number of infected animals in each of the groups as low as 83%.
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