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Planta Med 2016; 82(13): 1192-1201
DOI: 10.1055/s-0042-110323
Pharmacokinetic Investigations
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Caco-2 Permeability Studies and In Vitro hERG Liability Assessment of Tryptanthrin and Indolinone

Evelyn A. Jähne
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
,
Daniela E. Eigenmann
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
,
Fahimeh Moradi-Afrapoli
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
,
Sheela Verjee
2   Institute for Pharma Technology, School of Life Sciences, University of Applied Sciences Northwestern Switzerland, Muttenz, Switzerland
,
Veronika Butterweck
2   Institute for Pharma Technology, School of Life Sciences, University of Applied Sciences Northwestern Switzerland, Muttenz, Switzerland
,
Simon Hebeisen
3   BʼSYS GmbH, The Ionchannel Company, Witterswil, Switzerland
,
Timm Hettich
4   Institute for Chemistry and Bioanalytics, School of Life Sciences, University of Applied Sciences, Northwestern Switzerland, Muttenz, Switzerland
,
Götz Schlotterbeck
4   Institute for Chemistry and Bioanalytics, School of Life Sciences, University of Applied Sciences, Northwestern Switzerland, Muttenz, Switzerland
,
Martin Smieško
5   Molecular Modeling, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
,
Matthias Hamburger
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
,
Mouhssin Oufir
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
› Author Affiliations
Further Information

Publication History

received 23 March 2016
revised 27 May 2016

accepted 07 June 2016

Publication Date:
15 July 2016 (online)

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Abstract

Tryptanthrin and (E,Z)-3-(4-hydroxy-3,5-dimethoxybenzylidene)indolinone (indolinone) were recently isolated from Isatis tinctoria as potent anti-inflammatory and antiallergic alkaloids, and shown to inhibit COX-2, 5-LOX catalyzed leukotriene synthesis, and mast cell degranulation at low µM to nM concentrations. To assess their suitability for oral administration, we screened the compounds in an in vitro intestinal permeability assay using human colonic adenocarcinoma cells. For exact quantification of the compounds, validated UPLC-MS/MS methods were used. Tryptanthrin displayed high permeability (apparent permeability coefficient > 32.0 × 10−6 cm/s) across the cell monolayer. The efflux ratio below 2 (< 1.12) and unchanged apparent permeability coefficient values in the presence of the P-glycoprotein inhibitor verapamil (50 µM) indicated that tryptanthrin was not involved in P-glycoprotein interactions. For indolinone, a low recovery was found in the human colon adenocarcinoma cell assay. High-resolution mass spectrometry pointed to extensive phase II metabolism of indolinone (sulfation and glucuronidation). Possible cardiotoxic liability of the compounds was assessed in vitro by measurement of an inhibitory effect on human ether-a-go-go-related gene tail currents in stably transfected HEK 293 cells using the patch clamp technique. Low human ether-a-go-go-related gene inhibition was found for tryptanthrin (IC50 > 10 µM) and indolinone (IC50 of 24.96 µM). The analysis of compounds using various in silico methods confirmed favorable pharmacokinetic properties, as well as a slight inhibition of the human ether-a-go-go-related gene potassium channel at micromolar concentrations.

Key words

Caco-2 - metabolism - U(H)PLC-MS/MS - validation - HR-MS - hERG - Isatis tinctoria

Supporting Information

 

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