TY - JOUR
T1 - OLT1177, a β-sulfonyl nitrile compound, safe in humans, inhibits the NLRP3 inflammasome and reverses the metabolic cost of inflammation
AU - Marchetti, Carlo
AU - Swartzwelter, Benjamin
AU - Gamboni, Fabia
AU - Neff, Charles P
AU - Richter, Katrin
AU - Azam, Tania
AU - Carta, Sonia
AU - Tengesdal, Isak
AU - Nemkov, Travis
AU - D'Alessandro, Angelo
AU - Henry, Curtis
AU - Jones, Gerald S
AU - Goodrich, Scott A
AU - St Laurent, Joseph P
AU - Jones, Terry M
AU - Scribner, Curtis L
AU - Barrow, Robert B
AU - Altman, Roy D
AU - Skouras, Damaris B
AU - Gattorno, Marco
AU - Grau, Veronika
AU - Janciauskiene, Sabina
AU - Rubartelli, Anna
AU - Joosten, Leo A B
AU - Dinarello, Charles A
N1 - Copyright © 2018 the Author(s). Published by PNAS.
PY - 2018/2/13
Y1 - 2018/2/13
N2 - Activation of the NLRP3 inflammasome induces maturation of IL-1β and IL-18, both validated targets for treating acute and chronic inflammatory diseases. Here, we demonstrate that OLT1177, an orally active β-sulfonyl nitrile molecule, inhibits activation of the NLRP3 inflammasome. In vitro, nanomolar concentrations of OLT1177 reduced IL-1β and IL-18 release following canonical and noncanonical NLRP3 inflammasome activation. The molecule showed no effect on the NLRC4 and AIM2 inflammasomes, suggesting specificity for NLRP3. In LPS-stimulated human blood-derived macrophages, OLT1177 decreased IL-1β levels by 60% and IL-18 by 70% at concentrations 100-fold lower in vitro than plasma concentrations safely reached in humans. OLT1177 also reduced IL-1β release and caspase-1 activity in freshly obtained human blood neutrophils. In monocytes isolated from patients with cryopyrin-associated periodic syndrome (CAPS), OLT1177 inhibited LPS-induced IL-1β release by 84% and 36%. Immunoprecipitation and FRET analysis demonstrated that OLT1177 prevented NLRP3-ASC, as well as NLRP3-caspase-1 interaction, thus inhibiting NLRP3 inflammasome oligomerization. In a cell-free assay, OLT1177 reduced ATPase activity of recombinant NLRP3, suggesting direct targeting of NLRP3. Mechanistically, OLT1177 did not affect potassium efflux, gene expression, or synthesis of the IL-1β precursor. Steady-state levels of phosphorylated NF-κB and IkB kinase were significantly lowered in spleen cells from OLT1177-treated mice. We observed reduced IL-1β content in tissue homogenates, limited oxidative stress, and increased muscle oxidative metabolism in OLT1177-treated mice challenged with LPS. Healthy humans receiving 1,000 mg of OLT1177 daily for 8 d exhibited neither adverse effects nor biochemical or hematological changes.
AB - Activation of the NLRP3 inflammasome induces maturation of IL-1β and IL-18, both validated targets for treating acute and chronic inflammatory diseases. Here, we demonstrate that OLT1177, an orally active β-sulfonyl nitrile molecule, inhibits activation of the NLRP3 inflammasome. In vitro, nanomolar concentrations of OLT1177 reduced IL-1β and IL-18 release following canonical and noncanonical NLRP3 inflammasome activation. The molecule showed no effect on the NLRC4 and AIM2 inflammasomes, suggesting specificity for NLRP3. In LPS-stimulated human blood-derived macrophages, OLT1177 decreased IL-1β levels by 60% and IL-18 by 70% at concentrations 100-fold lower in vitro than plasma concentrations safely reached in humans. OLT1177 also reduced IL-1β release and caspase-1 activity in freshly obtained human blood neutrophils. In monocytes isolated from patients with cryopyrin-associated periodic syndrome (CAPS), OLT1177 inhibited LPS-induced IL-1β release by 84% and 36%. Immunoprecipitation and FRET analysis demonstrated that OLT1177 prevented NLRP3-ASC, as well as NLRP3-caspase-1 interaction, thus inhibiting NLRP3 inflammasome oligomerization. In a cell-free assay, OLT1177 reduced ATPase activity of recombinant NLRP3, suggesting direct targeting of NLRP3. Mechanistically, OLT1177 did not affect potassium efflux, gene expression, or synthesis of the IL-1β precursor. Steady-state levels of phosphorylated NF-κB and IkB kinase were significantly lowered in spleen cells from OLT1177-treated mice. We observed reduced IL-1β content in tissue homogenates, limited oxidative stress, and increased muscle oxidative metabolism in OLT1177-treated mice challenged with LPS. Healthy humans receiving 1,000 mg of OLT1177 daily for 8 d exhibited neither adverse effects nor biochemical or hematological changes.
KW - Animals
KW - Anti-Inflammatory Agents/chemistry
KW - Caspase 1/metabolism
KW - Cells, Cultured
KW - Humans
KW - Inflammasomes/antagonists & inhibitors
KW - Inflammation/chemically induced
KW - Interleukin-18/metabolism
KW - Interleukin-1beta/metabolism
KW - Lipopolysaccharides/toxicity
KW - Macrophages/drug effects
KW - Male
KW - Mice
KW - Mice, Inbred C57BL
KW - NLR Family, Pyrin Domain-Containing 3 Protein/genetics
KW - Nitriles/chemistry
U2 - 10.1073/pnas.1716095115
DO - 10.1073/pnas.1716095115
M3 - Article
C2 - 29378952
VL - 115
SP - E1530-E1539
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 7
ER -