Pharmacodynamics and biomarker analyses
inflammation, pre‐dose, SD
BLOOD, INFLAMMATION
Secreted IL‐1β was assessed in whole blood following ex vivo stimulation with LPS, to evaluate target engagement with the NLRP3 inflammasome. Data were expressed as percentage of inhibition of IL‐1β release compared to baseline (Figure Inhibition of IL‐1β release in whole blood upon ex vivo stimulation with LPS. BL, baseline; d, days. Blood samples were obtained from patients treated with either placebo or selnoflast, at various timepoints pre‐ and post‐dosing. Blood samples were treated with LPS for 24 h prior to measurement of IL‐1β levels (in the supernatants). The level of IL‐1β is expressed as the percentage change from baseline (pre‐dose), including standard error bars.We also evaluated blood, sigmoid colon tissue and stool in order to understand the effect of selnoflast administration on disease‐relevant biomarkers (Figure Effect of selnoflast on markers of inflammation in blood, sigmoid colon tissue and stool. The graph depicts the logSigmoid colon tissue biopsies were assessed histologically before and after treatment (baseline and Day 7) using the standard disease‐specific histological indices Geboes score and Nancy index. No notable differences were observed in either of the two indices between the study arms (results not shown). Three out of the 13 selnoflast‐treated patients showed a marked reduction from baseline in the level of neutrophils in sigmoid colon tissue, contributing to an overall modest mean (± SD) change from baseline of −13.23 (± 140.76) cells/mmStool calprotectin levels after treatment were higher in the selnoflast arm: on Day 7, the mean (± SD) change from baseline was +4146.6 (± 12321.12) μg/g, compared to a reduction of 7351.70 (± 14111.83) μg/g in the placebo arm. The difference was mainly driven by two individuals, one in each arm. One patient in the placebo arm showed a drop in fecal calprotectin from 29947 μg/g at baseline to 1562 μg/g on Day 7. One patient in the selnoflast arm showed an increase in fecal calprotectin from 1553 μg/g at baseline to 39770 μg/g at Day 7. It should also be noted that a substantial proportion (8/19) of the baseline stool samples were not taken pre‐biopsy as per protocol, rendering the results for this biomarker uninterpretable.We assessed the expression of an IL‐1 gene signature (as a downstream transcriptomic marker in lieu of tissue IL‐1β where no validated assay was available to assess the active form) in sigmoid colon tissue by qPCR before and after treatment. Expression levels were normalized to a set of housekeeping genes and the logExpression of an IL‐1 gene signature in sigmoid colon tissue. RNA was extracted from sigmoid colon tissue samples that were obtained before treatment and on Day 7 to evaluate the expression of an IL‐1 gene signature by quantitative Polymerase Chain Reaction. The gene signature consisted of a panel of 8 genes: CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CCL2, IL6 and TNFAIP6. The graph depicts the log