Novel NLRP3 inhibitor INF195: Low doses provide effective protection against myocardial ischemia/reperfusion injury

  • Vascul Pharmacol. 2024 Jun 17:156:107397. doi: 10.1016/j.vph.2024.107397.
Simone Gastaldi  1 ,  Magalì Giordano  2 ,  Federica Blua  1 ,  Chiara Rubeo  2 ,  Valentina Boscaro  1 ,  Saveria Femminò  2 ,  Stefano Comità  2 ,  Eleonora Gianquinto  1 ,  Vanessa Landolfi  2 ,  Elisabetta Marini  1 ,  Margherita Gallicchio  1 ,  Francesca Spyrakis  1 ,  Pasquale Pagliaro  3 ,  Massimo Bertinaria  4 ,  Claudia Penna  5
Affiliations
  • 1. Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, 10125 Torino, Italy.
  • 2. Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, Orbassano, 10043 Torino, Italy.
  • 3. Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, Orbassano, 10043 Torino, Italy; INRC, Bologna, Italy. Electronic address: [email protected].
  • 4. Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, 10125 Torino, Italy; Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, Orbassano, 10043 Torino, Italy. Electronic address: [email protected].
  • 5. Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, Orbassano, 10043 Torino, Italy; INRC, Bologna, Italy.
PMID: 38897555 DOI: 10.1016/j.vph.2024.107397
Abstract

Background: Several factors contribute to ischemia/reperfusion injury (IRI), including activation of the NLRP3 inflammasome and its byproducts, such as interleukin-1β (IL-1β) and Caspase-1. However, NLRP3 may paradoxically exhibit cardioprotective properties. This study aimed to assess the protective effects of the novel NLRP3 Inhibitor, INF195, both in vitro and ex vivo.

Methods: To investigate the relationship between NLRP3 and myocardial IRI, we synthetized a series of novel NLRP3 inhibitors, and investigated their putative binding mode via docking studies. Through in vitro studies we identified INF195 as optimal for NLRP3 inhibition. We measured infarct-size in isolated mouse hearts subjected to 30-min global ischemia/one-hour reperfusion in the presence of three different doses of INF195 (5, 10, or 20-μM). We analyzed Caspase-1 and IL-1β concentration in cardiac tissue homogenates by ELISA. Statistical significance was determined using one-way ANOVA followed by Tukey's test.

Results and conclusion: INF195 reduces NLRP3-induced Pyroptosis in human macrophages. Heart pre-treatment with 5 and 10-μM INF195 significantly reduces both infarct size and IL-1β levels. Data suggest that intracardiac NLRP3 activation contributes to IRI and that low doses of INF195 exert cardioprotective effects by reducing infarct size. However, at 20-μM, INF195 efficacy declines, leading to a lack of cardioprotection. Research is required to determine if high doses of INF195 have off-target effects or dual roles, potentially eliminating both harmful and cardioprotective functions of NLRP3. Our findings highlight the potential of a new chemical scaffold, amenable to further optimization, to provide NLRP3 inhibition and cardioprotection in the ischemia/reperfusion setting.

Keywords
Cardioprotection; Interleukin-1β; NLRP3 inflammasome.
Products