Identification, Synthesis, and Biological Evaluation of the Major Human Metabolite of NLRP3 Inflammasonne Inhibitor MCC950

• Published in: ACS medicinal chemistry letters Vol. 7; no. 12; pp. 1034 - 1038
• Main Authors: Salla, Manohar, Butler, Mark S., Pelingon, Ruby, Kaeslin, Geraldine, Croker, Daniel E., Reid, Janet C., Baek, Jong Min, Bernhardt, Paul V., Gillam, Elizabeth M. J., Cooper, Matthew A., Robertson, Avril A. B.
• Format: Journal Article
• Language: English
• Published: WASHINGTON Amer Chemical Soc 01.12.2016
• Subjects: Chemistry, Medicinal; Life Sciences & Biomedicine; Pharmacology & Pharmacy; Science & Technology; MCC950; metabolite; inflammasome; DRUG DISCOVERY; DISEASE; NLRP3; microsome; cytochrome P450
• ISSN: 1948-5875; 1948-5875
• DOI: 10.1021/acsmedchemlett.6b00198

MCC950 is an orally bioavailable small molecule inhibitor of the NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome that exhibits remarkable activity in multiple models of inflammatory disease. Incubation of MCC950 with human liver microsomes, and subsequent analysis by HPLC-MS/MS, revealed a major metabolite, where hydroxylation of MCC950 had occurred on the 1,2,3,5,6,7-hexahydro-s-indacene moiety. Three possible regioisomers were synthesized, and coelution using HPLC-MS/MS confirmed the structure of the metabolite. Further synthesis of individual enantiomers and coelution studies using a chiral column in HPLC-MS/MS showed the metabolite was R-(+)- N-((1-hydroxy-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoy1)-4-(2-hydroxypropan-2-y0furan-2-sulfonamide (2a). Incubation of MCC950 with a panel of cytochrome P450 enzymes showed P450s 2A6, 2C9, 2C18, 2C19, 2J2, and 3A4 catalyze the formation of the major metabolite 2a, with a lower level of activity shown by P450s 1A2 and 2B6. All of the synthesized compounds were tested for inhibition of NLRP3-induced production of the pro-inflammatory cytokine IL-1 beta from human monocyte derived macrophages. The identified metabolite 2a was 170-fold less potent than MCC950, while one regioisomer had nanomolar inhibitory activity. These findings also give first insight into the SAR of the hexahydroindacene moiety.