title: Physalins with anti-inflammatory activity are present in Physalis alkekengi var. franchetii and can function as Michael reaction acceptors.
abstract: Michael reaction acceptors (MRAs) are a class of active molecules that are directly or indirectly involved in various cellular processes, including the regulation of many signaling pathways. In this study, the inducible nitric oxide synthase (iNOS) assay was used to demonstrate that the dichloromethane extract of Physalis alkekengi var. franchetii (DCEP) possesses anti-inflammatory activity that might be attributed to the modification of key cysteine residues in IKKβ by the MRAs in DCEP. To isolate these MRAs, glutathione (GSH) was employed, and a simple ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) screening method was developed to investigate the GSH conjugates with potential MRAs. Five physalins, including one new compound isophysalin A (2), together with four known steroidal compounds, physalin A (1), physalin O (3), physalin L (4) and physalin G (5), were isolated to evaluate the GSH conjugating abilities, and it was indicated that compounds 1, 2 and 3, which had a common α,β-unsaturated ketone moiety, exhibited conjugating abilities with GSH and also showed significant nitric oxide (NO) production inhibiting activities. The anti-inflammatory activities of compounds 1, 2 and 3 might be attributed to their targeting multiple cysteine residues on IKKβ; therefore, the alkylation of IKKβ by compound 1 was further studied by micrOTOF-MS. The result showed that six cysteine residues (C(59), C(179), C(299), C(370), C(412), and C(618)) were alkylated, which indicated that IKKβ is a potential target for the anti-inflammatory activity of physalin A.
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus, No. 866 Yuhangtang Road, Hangzhou 310058, PR China.
authors.forename: Long
authors.initials: L
authors.lastname: Ji
authors.forename: Yonglei
authors.initials: Y
authors.lastname: Yuan
authors.forename: Liping
authors.initials: L
authors.lastname: Luo
authors.forename: Zhe
authors.initials: Z
authors.lastname: Chen
authors.forename: Xiaoqiong
authors.initials: X
authors.lastname: Ma
authors.forename: Zhongjun
authors.initials: Z
authors.lastname: Ma
authors.forename: Lin
authors.initials: L
authors.lastname: Cheng
mesh_terms: D000478:Alkylation / Q000187:drug effects; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000378:metabolism; D005978:Glutathione / Q000737:chemistry; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D015394:Molecular Structure; D009569:Nitric Oxide / Q000378:metabolism; D032321:Physalis / Q000737:chemistry*; D010840:Phytosterols / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D012632:Secosteroids / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D054358:Withanolides
chemical_list: D000893:Anti-Inflammatory Agents; D008070:Lipopolysaccharides; D010840:Phytosterols; D012632:Secosteroids; D054358:Withanolides; C574201:physalin A; C569084:physalin G; C574203:physalin L; C574202:physalin O; D009569:Nitric Oxide; D051550:I-kappa B Kinase; D005978:Glutathione; D003545:Cysteine