title: Identification of Eupatilin from Artemisia argyi as a Selective PPARα Agonist Using Affinity Selection Ultrafiltration LC-MS.
abstract: Peroxisome proliferator-activated receptors (PPARs) are key nuclear receptors and therapeutic targets for the treatment of metabolic diseases through the regulation of insulin resistance, diabetes, and dyslipidemia. Although a few drugs that target PPARs have been approved, more diverse and novel PPAR ligands are necessary to improve the safety and efficacy of available drugs. To expedite the search for new natural agonists of PPARs, we developed a screening assay based on ultrafiltration liquid chromatography-mass spectrometry (LC-MS) that is compatible with complex samples such as dietary foods or botanical extracts. The known PPARα and/or PPARγ ligands resveratrol and rosiglitazone were used as positive controls to validate the developed method. When applied to the screening of an Artemisia argyi extract, eupatilin was identified as a selective PPARα ligand. A PPAR competitive binding assay based on FRET detection also confirmed eupatilin as a selective PPARα agonist exhibiting a binding affinity of 1.18 μM (IC50). Furthermore, eupatilin activation of the transcriptional activity of PPARα was confirmed using a cell-based transactivation assay. Thus, ultrafiltration LC-MS is a suitable assay for the identification of PPAR ligands in complex matrixes such as extracts of dietary foods and botanicals.
authors.affiliation: Laboratory of Biomodulation, Natural Products Research Institute, Korea Institute of Science and Technology, Gangneung 210-340, Korea. yongsoo.choi@kist.re.kr.
authors.forename: Yongsoo
authors.initials: Y
authors.lastname: Choi
authors.affiliation: Laboratory of Natural Skinomics, Natural Products Research Institute, Korea Institute of Science and Technology, 290 Daejeon-dong, Gangneung-si, Gangwon-do 210-340, Korea. yj0103@kist.re.kr.
authors.forename: Yujung
authors.initials: Y
authors.lastname: Jung
authors.affiliation: Laboratory of Natural Skinomics, Natural Products Research Institute, Korea Institute of Science and Technology, 290 Daejeon-dong, Gangneung-si, Gangwon-do 210-340, Korea. snkim@kist.re.kr.
authors.forename: Su-Nam
authors.initials: SN
authors.lastname: Kim
mesh_terms: D019306:Artemisia / Q000737:chemistry*; D002460:Cell Line; D005419:Flavonoids / Q000737:chemistry / Q000302:isolation & purification / Q000493:pharmacokinetics / Q000494:pharmacology; D006801:Humans; D047493:PPAR alpha / Q000819:agonists / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D015533:Transcriptional Activation / Q000187:drug effects*
chemical_list: D005419:Flavonoids; D047493:PPAR alpha; C045325:eupatilin
keywords: Artemisia argyi; PPARα; eupatilin; ultrafiltration LC-MS