2. Academic Validation
  3. Novel NLRP3 inhibitor INF195: Low doses provide effective protection against myocardial ischemia/reperfusion injury

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

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: pasquale.pagliaro@unito.it.
  • 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: massimo.bertinaria@unito.it.
  • 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.

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