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. Author manuscript; available in PMC: 2019 Nov 15.
Published in final edited form as: J Med Chem. 2019 Oct 31;62(21):9718–9731. doi: 10.1021/acs.jmedchem.9b01155

Figure 6.

Figure 6.

Compound 17 selectively inhibits the NLRP3 inflammasome. (A) Mouse peritoneal macrophages were primed with LPS (1 μg/mL) for 4.5 h and then treated with indicated compounds at indicated concentrations when adding ATP (5 mM) stimulation for 30 min. IL-β in the culture media was assayed by ELISA. (B) J774A. 1 cells were treated with LPS (1 μg/mL) and 17 (10 μM) for 1 h. Flagellin (1 μg/mL) was added and allowed to incubate for 6 hr or (Poly(dA:dT)) (4 μg/ml) for 8 hr. The supernatants were collected and levels of IL-1β were measured by ELISA. Serum levels of IL-1β.(C) and TNF-α (D) from C57BL/6 (n=4 per group) mice pretreated 17 (10 mg/kg) or MCC950 (10 mg/kg) were measured by ELISA 2.5 h after i.p. injection of LPS (50 mg/kg). (E) Serum levels of IL-1β and TNF-α under indicated treatment conditions (both 17 and MCC950 were tested at 10 mg/kg) of nlrp3−/− mice (n=3 per group) were measured by ELISA 2.5 h after i.p. injection of LPS (25 mg/kg). For in vitro assasys, data are expressed as mean ± SEM from at least 3 independent experiments with at least triplicates for each experiment. For in vivo assays, data are expressed as mean ± SD. Statistical analysis by student t-test.