title: Interaction of six protoberberine alkaloids with human organic cation transporters 1, 2 and 3.
abstract: 1. Organic cation transporters (OCTs) play an important role in drug safety and efficacy. Protoberberine alkaloids are ubiquitous organic cations or weak bases with remarkable biological actives. This study was to elucidate the potential interaction of alkaloids (coptisine, jatrorrhizine, epiberberine, berberrubine, palmatine and corydaline) with OCTs using Madin-Darby canine kidney (MDCK) cells stably expressing human OCT1, OCT2 and OCT3. 2. All the tested alkaloids significantly inhibited the uptake of MPP(+), a model OCT substrate, in MDCK-hOCTs cells with the IC50 of 0.931-9.65 μM. Additionally, coptisine, jatrorrhizine and epiberberine were substrates of all the hOCTs with the Km of 0.273-5.80 μM, whereas berberrubine was a substrate for hOCT1 and hOCT2, but not for hOCT3, the Km values were 1.27 and 1.66 μM, respectively. The transport capacity of coptisine in MDCK cells expressing the variants of hOCT1-P341L or hOCT2-A270S was significantly higher than that in wild-type (WT) cells with the Clint (Vmax/Km) of 379 ± 7.4 and 433 ± 5.7 μl/mg protein/min, respectively. 3. The above data indicate that the tested alkaloids are potent inhibitors, and coptisine, jatrorrhizine, epiberberine and berberrubine are substrates of hOCT1, hOCT2 and/or hOCT3 with high affinity. In addition, the variants (OCT1-P341L and OCT2-A270S) possess higher transport capacity to coptisine than WT hOCTs.
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Liping
authors.initials: L
authors.lastname: Li
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Siyuan
authors.initials: S
authors.lastname: Sun
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Yayun
authors.initials: Y
authors.lastname: Weng
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Feifeng
authors.initials: F
authors.lastname: Song
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Sisi
authors.initials: S
authors.lastname: Zhou
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Mengru
authors.initials: M
authors.lastname: Bai
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhou
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Su
authors.initials: S
authors.lastname: Zeng
authors.affiliation: a Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , Zhejiang , P.R. China.
authors.forename: Huidi
authors.initials: H
authors.lastname: Jiang
mesh_terms: D015655:1-Methyl-4-phenylpyridinium / Q000493:pharmacokinetics; D000818:Animals; D001599:Berberine / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D001600:Berberine Alkaloids / Q000493:pharmacokinetics*; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D004285:Dogs; D006801:Humans; D020128:Inhibitory Concentration 50; D061985:Madin Darby Canine Kidney Cells; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D027701:Organic Cation Transport Proteins / Q000235:genetics / Q000378:metabolism*; D027702:Organic Cation Transporter 1 / Q000235:genetics / Q000378:metabolism*; D000075102:Organic Cation Transporter 2; D011110:Polymorphism, Genetic; D053719:Tandem Mass Spectrometry
chemical_list: D001600:Berberine Alkaloids; D050814:Octamer Transcription Factor-3; D027701:Organic Cation Transport Proteins; D027702:Organic Cation Transporter 1; D000075102:Organic Cation Transporter 2; C494173:POU5F1 protein, human; C491013:SLC22A2 protein, human; C452799:corydaline; C055785:jatrorrhizine; C034384:coptisine; D001599:Berberine; C115958:berberrubine; C009090:protoberberine; C061432:epiberberine; C005413:palmatine; D015655:1-Methyl-4-phenylpyridinium
keywords: Cells; inhibitor; interaction; organic cation transporters; protoberberine alkaloids; substrate