OLT1177, a ss-sulfonyl nitrile compound, safe in humans, inhibits the NLRP3 inflammasome and reverses the metabolic cost of inflammation

• Type of Material: Article
• Authors: Carlo Marchetti, University of Colorado Denver; Benjamin Swartzwelter, University of Colorado Denver; Fabia Gamboni, University of Colorado Denver; Charles P. Neff, University of Colorado Denver; Katrin Richter, Justus Liebig University Giessen; Tania Azam, University of Colorado Denver; Sonia Carta, Istituto Nazionale per la Ricerca sul Cancro; Isak Tengesdal, University of Colorado Denver; Travis Nemkov, University of Colorado; Angelo D'Alessandro, University of Colorado; Curtis Henry, Emory University; Gerald S. Jones, Chemic Laboratories, Inc.; Scott A. Goodrich, Chemic Laboratories, Inc.; Joseph P. St. Laurent, Chemic Laboratories, Inc.; Terry M. Jones, J&S Studies, Inc.; Curtis L. Scribner, Olatec Therapeutics LLC; Robert B. Barrow, Olatec Therapeutics LLC; Roy D. Altman, University of California Los Angeles; Damaris B. Skouras, Olatec Therapeutics LLC; Marco Gattorno, G. Gaslini Institute; Veronika Grau, Justus-Liebig-University Giessen; Sabina Janciauskiene, Hannover Medical School; Anna Rubartelli, Istituto Nazionale per la Ricerca sul Cancro; Leo A. B. Joosten, Radboud University Nijmegen; Charles A. Dinarello, University of Colorado Denver
• Language: English
• Date: 2018-02-13
• Publisher: National Academy of Sciences
• Publication Version: Final Published Version
• Copyright Statement: © 2018 the Author(s). Published by PNAS.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1073/pnas.1716095115
• Title of Journal or Parent Work: Proceedings of the National Academy of Sciences
• ISSN: 0027-8424
• Volume: 115
• Issue: 7
• Start Page: E1530
• End Page: E1539
• Grant/Funding Information: These studies are supported by NIH Grant AI-15614, The Interleukin Foundation, Olatec Therapeutics LLC, and German Research Foundation Grant GR 1094/7-1.
• Keywords: interleukin-1; GOUTY-ARTHRITIS; KAPPA-B; KINASES; INTERLEUKIN-1; NLRP3; NEUTROPHILS; DISEASE; DENDRITIC CELL ACTIVATION; Science & Technology; SKELETAL-MUSCLE; Science & Technology - Other Topics; caspase-1; GLUCOSE-METABOLISM; FAMILIAL MEDITERRANEAN FEVER; Multidisciplinary Sciences
• Research Categories: Health Sciences, Medicine and Surgery

Abstract

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.