Identification, synthesis, and biological evaluation of the major human metabolite of NLRP3 inflammasome inhibitor MCC950

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. and Robertson, Avril A. B. (2016) Identification, synthesis, and biological evaluation of the major human metabolite of NLRP3 inflammasome inhibitor MCC950. ACS Medicinal Chemistry Letters, 7 12: 1034-1038. doi:10.1021/acsmedchemlett.6b00198


Author 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.
Title Identification, synthesis, and biological evaluation of the major human metabolite of NLRP3 inflammasome inhibitor MCC950
Journal name ACS Medicinal Chemistry Letters   Check publisher's open access policy
ISSN 1948-5875
Publication date 2016-09-27
Year available 2016
Sub-type Article (original research)
DOI 10.1021/acsmedchemlett.6b00198
Open Access Status Not yet assessed
Volume 7
Issue 12
Start page 1034
End page 1038
Total pages 5
Place of publication Washington, DC, United States
Publisher American Chemical Society
Language eng
Abstract 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.
Formatted abstract
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)carbamoyl)-4-(2-hydroxypropan-2-yl)furan-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-1β 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.
Keyword NLRP3
Inflammasome
MCC950
Microsome
Metabolite
Cytochrome P450
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID APP1086786
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
School of Chemistry and Molecular Biosciences
Institute for Molecular Bioscience - Publications
 
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Created: Wed, 04 Jan 2017, 00:47:43 EST by Susan Allen on behalf of Institute for Molecular Bioscience