Impact of chemical modifications in vivo and activity–duration relationship. (A) Compounds demonstrating a significant impact of chemical modifications on Day 2, compared with unmodified parent, at Day 1 were categorized accordingly. Categorization is based on the direction of significant comparisons (p < 0.05) as determined by two-way ANOVA (sequence × modification) of log-transformed ED50 data. Bars are additive. (B) Compounds showing a significant difference compared with unmodified parent compound on Day 9, Day 21, or both were categorized accordingly. Categorization is based on direction of significant comparisons (p < 0.05) as determined by two-way ANOVA (time × modification) of single dose data. Bars are additive. FVII (C), and PTEN (D) T50 data from all canonical (black) and dicer-substrate (gray) compounds were plotted against corresponding LogED50 values. A linear regression was performed to describe the relationship between these measures of activity and duration. Data points represent n = 3–5, with two technical replicates. Modifications are (UU) unmodified/unmodified, (HL) heavy/light, (HA) heavy/alternating, and (UA) unmodified/alternating.
