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Planta Med 2017; 83(14/15): 1169-1175
DOI: 10.1055/s-0043-110768
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

HPLC-Based Activity Profiling for GABAA Receptor Modulators in Searsia pyroides Using a Larval Zebrafish Locomotor Assay[*]

Fahimeh Moradi-Afrapoli
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
,
Hannes van der Merwe
2   Faculty of Natural and Agricultural Sciences, University of the Free State, Bloemfontein, Republic of South Africa
,
Maria De Mieri
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
,
Anke Wilhelm
2   Faculty of Natural and Agricultural Sciences, University of the Free State, Bloemfontein, Republic of South Africa
,
Marco Stadler
3   Institute of Pharmacology and Toxicology, Pharmaziezentrum, University of Vienna, Vienna, Austria
,
Pieter C. Zietsman
4   National Museum, Bloemfontein, Republic of South Africa
,
Steffen Hering
3   Institute of Pharmacology and Toxicology, Pharmaziezentrum, University of Vienna, Vienna, Austria
,
Kenneth Swart
2   Faculty of Natural and Agricultural Sciences, University of the Free State, Bloemfontein, Republic of South Africa
5   FARMOVS-PAREXEL, University of the Free State, Bloemfontein, South Africa
,
Matthias Hamburger
1   Pharmaceutical Biology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland
› Author Affiliations
Further Information

Publication History

received 20 February 2017
revised 04 May 2017

accepted 04 May 2017

Publication Date:
16 May 2017 (online)

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Abstract

A dichloromethane extract from leaves of Searsia pyroides potentiated gamma aminobutyric acid-induced chloride currents by 171.8 ± 54% when tested at 100 µg/mL in Xenopus oocytes transiently expressing gamma aminobutyric acid type A receptors composed of α 1 β 2 γ 2s subunits. In zebrafish larvae, the extract significantly lowered pentylenetetrazol-provoked locomotion when tested at 4 µg/mL. Active compounds of the extract were tracked with the aid of HPLC-based activity profiling utilizing a previously validated zebrafish larval locomotor activity assay. From two active HPLC fractions, compounds 1 – 3 were isolated. Structurally related compounds 4 – 6 were purified from a later eluting inactive HPLC fraction. With the aid of 1H and 13C NMR and high-resolution mass spectrometry, compounds 1 – 6 were identified as analogues of anacardic acid. Compounds 1 – 3 led to a concentration-dependent decrease of pentylenetetrazol-provoked locomotion in the zebrafish larvae model, while 4 – 6 were inactive. Compounds 1 – 3 enhanced gamma aminobutyric acid-induced chloride currents in Xenopus oocytes in a concentration-dependent manner, while 4 – 6 only showed marginal enhancements of gamma aminobutyric acid-induced chloride currents. Compounds 2, 3, and 5 have not been reported previously.

Key words

Searsia pyroides - Anacardiaceae - GABAA receptor - zebrafish larvae - Xenopus oocytes - pentylenetetrazole - HPLC-based activity profiling

* Dedicated to Professor Dr. Max Wichtl in recognition of his outstanding contribution to pharmacognosy research.


Supporting Information

    1H-NMR data, 13C-NMR data (extracted from HSQC-DEPT and HMBC spectra for compounds 1 and 3 – 6), HR ESIMS data of 1 – 6, and 13C and 1H-NMR shifts for E and Z isomers of compound 1 (DMSO-d 6) calculated with ACDLabs software are available as Supporting Information.

  • Supporting Information
 

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