Design, Synthesis, and Evaluation of Acrylamide Derivatives as Direct NLRP3 Inflammasome Inhibitors

Abstract

NLRP3 inflammasome plays a key role in the intracellular activation of caspase-1, processing of pro-inflammatory interleukin-1β (IL-1β), and pyroptotic cell death cascade. The overactivation of NLRP3 is implicated in the pathogenesis of autoinflammatory diseases, known as cryopyrin-associated periodic syndromes (CAPS), and in the progression of several diseases, such as atherosclerosis, type-2 diabetes, gout, and Alzheimer's disease. In this study, the synthesis of acrylamide derivatives and their pharmaco-toxicological evaluation as potential inhibitors of NLRP3-dependent events was undertaken. Five hits were identified and evaluated for their efficiency in inhibiting IL-1β release from different macrophage subtypes, including CAPS mutant macrophages. The most attractive hits were tested for their ability to inhibit NLRP3 ATPase activity on human recombinant NLRP3. This screening allowed the identification of 14, 2-(2-chlorobenzyl)-N-(4-sulfamoylphenethyl)acrylamide, which was able to concentration-dependently inhibit NLRP3 ATPase with an IC50 value of 74 μm. The putative binding pose of 14 in the ATPase domain of NLRP3 was also proposed.

Pharmacological Inhibition of NLRP3 Inflammasome Attenuates Myocardial Ischemia/Reperfusion Injury by Activation of RISK and Mitochondrial Pathways

“…Stock solution was then diluted at a final concentration of 50  µ M in the perfusion buffer (see below). The description of the synthesis and the specificity of the inhibitor is included in the Supplemental Material (available online at http://dx.doi.org/10.1155/2016/5271251), according to previous publications [16, 17]. …”

Therapeutic modulation of inflammasome pathways

“…It was shown to inhibit ATPase activity of human recombinant NLRP3 and IL-1β release from primary and immortalized macrophages. 134 Unlike the previously mentioned synthetic molecules, oridonin is a natural compound derived from the medicinal plant Rabdosia rubescens. It was shown to inhibit NLRP3 activation by preventing the interaction between NLRP3 and NEK7 through covalent binding to Cys279 in the NACHT domain of NLRP3 (Figure 3).…”

Targeting the Microglial NLRP3 Inflammasome and Its Role in Parkinson's Disease

“…A series of acrylate derivative compounds, IFN58 (IC 50 of 74 μM), IFN39 (IC 50 of 10 μM), and BOT‐4‐one (3 μM), a benzoxathiole derivative, inhibit ATPase activity of the NLRP3 inflammasome . BOT‐4‐one (3 μM) modulates ATPase activity of the NLRP3 inflammasome through enhancement of the ubiquitination level of NLRP3 …”

“…123 A series of acrylate derivative compounds, IFN58 (IC 50 of 74 μM), IFN39 (IC 50 of 10 μM), and BOT-4-one (3 μM), a benzoxathiole derivative, inhibit ATPase activity of the NLRP3 inflammasome. [124][125][126] BOT-4-one (3 μM) modulates ATPase activity of the NLRP3 inflammasome through enhancement of the ubiquitination level of NLRP3. [127][128][129] In addition, many promising compounds, such as 2-aminoethaxydiphenyl borate (2-APB), which belongs to a novel boron compound series, and Fc11a-2 and OLT1177, have been reported to inhibit the NLRP3 inflammasome and the release of proinflammatory cytokines.…”