Table 1.
Overview of compound potencies in multiple assays.
Fig 1.
Dose response curves for human and mouse ligand binding and functional assays.
Response vs concentration profile for compound 1–6 in the ligand binding assays (human = red, mouse = blue), the primary human Th17 cell assay (purple) and in the thymocyte apoptosis assay (orange). Error bars indicate standard deviation.
Fig 2.
Thymocyte numbers and exposure predictions and measurements after compound treatment in the thymus involution model.
A) Absolute numbers of CD4+CD8+ thymocytes in the thymus at termination after dosing BID for 48 hours with compound 3 B) and compound 4 in the thymus involution model in vivo. Significance levels: * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001). Exposure predictions (lines) and terminal concentrations (dots) in compound treated mice in the thymus involution model in vivo for C) compound 3 and D) compound 4.
Fig 3.
Reduction of IL-17A and inflammatory readouts in the ear of IMQ treated mice after treatment with compound 3.
Cell populations in the ear of IMQ treated animals after treatment with compound 3. A) Percentage of total IL-17A producing cells among CD45+ cells B) Percentage IL-17A producing cells among γδ T cells C) Percentage of γδ T cells among CD45+ cells D) Ear thickness measured over the course of the IMQ induces skin inflammation model in vivo. Significance levels: * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001).
Fig 4.
Correlation of screening assays with functional readouts show separation of IL-17A and thymocyte apoptosis effects.
A) Comparison of human and mouse inhibition constant (Ki) for RORγt B) Comparison of percentage effect for the compounds in the human ligand binding assay vs observed inhibition in the primary human Th17 cell assay at corresponding concentrations. C) Comparison of percentage effect for compounds in the mouse ligand binding assay vs observed inhibition in the mouse thymocyte apoptosis assay at corresponding concentrations. D) Comparison of percentage effect for compounds in the human IL-17 assay vs observed inhibition in the mouse thymocyte apoptosis assay at corresponding concentrations. The colour scales depict the fraction unbound (%) in FBS.