HPLC-Based Activity Profiling for GABA A Receptor Modulators in Murraya exotica

Abstract

A dichloromethane extract from twigs and leaves of Murraya exotica produced allosteric potentiation of gamma aminobutyric acid (GABA) induced chloride currents in a microelectrode assay in Xenopus laevis oocytes expressing GABA receptors of α₁, β₂, γ_2s subunit composition. The activity was tracked by HPLC-based activity profiling utilizing a zebrafish locomotor activity assay. Osthol (9) was identified as the main active compound. In addition, five other coumarins and four flavonols were identified. Osthol (9) and structurally related coumurrayin (10) were tested in the Xenopus oocyte assay. Compound 9 potentiated GABA_A-induced chloride currents by 487 ± 42%, with an EC₅0 of 46 ± 10 μM, while 10 showed negligible effects on chloride currents. In silico evaluation of physicochemical properties showed that 9 and 10 had properties that are favorable for oral bioavailability and BBB permeability.

Keywords

Murraya exotica Zebrafish larvae HPLC-based activity profiling Osthol

References

Olsen

, Sieghart

(2008) International Union of Pharmacology. LXX. Subtypes of γ-aminobutyric acid_A receptors: classification on the basis of subunit composition, pharmacology, and function. Update. Pharmacological Reviews, 60, 243-260.

Simon

, Wakimoto

, Fujita

, Lalande

, Barnard

. (2004) Analysis of the set of GABA_A receptor genes in the human genome. The Journal of Biological Chemistry, 40, 41422-41435.

Uusi-Oukari

, Korpi

. (2010) Regulation of GABA_A receptor subunit expression by pharmacological agents. Pharmacological Review, 62, 97-135.

Sieghart

(2015) Allosteric modulation of GABA_A receptors via multiple drug-binding sites. Advances in Pharmacology, 23, 53-96.

Khom

, Baburin

, Timin

, Hohaus

, Trauner

, Kopp

, Hering

(2007) Valerenic acid potentiates and inhibits GABA_A receptors: molecular mechanism and subunit specificity. Neuropharmacology, 53, 178-187.

Zaugg

, Baburin

, Strommer

, Kim

H-J

, Hering

, Hamburger

(2010) HPLC-based activity profiling: discovery of piperine as a positive GABA receptor modulator targeting a benzodiazepine-independent binding site. Journal of Natural Products, 73, 185-191.

Kim

, Baburin

, Khom

, Hering

, Hamburger

(2008) HPLC-Based activity profiling approach for the discovery of GABA_A receptor ligands using an automated two microelectrode voltage clamp assay on Xenopus oocytes. Planta Medica, 74, 521-526.

Moradi-Afrapoli

, Ebrahimi

, Smiesko

, Hamburger

(2017) HPLC-based activity profiling for GABA_A modulators in extract: Validation of an approach utilizing a larval zebrafish locomotor assay. Journal of Natural Products, 80, 1548-1557.

Baraban

, Taylor

, Castro

, Baier

(2005) Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression. Neuroscience, 131, 759-768.

10.

Jeong

J-Y

, Kwon

H-B

, Ahn

J-C

, Kang

, Kwon

S-H

, Park

, Kim

K-W.

(2008) Functional and developmental analysis of the blood-brain barrier in zebrafish. Brain Research Bulletin, 75, 619-628.

11.

Fleming

, Diekmann

, Goldsmith

(2013) Functional characterization of the maturation of the blood-brain barrier in larval zebrafish. Plos One, 8, 1-12.

12.

Lipinski

, Lombardo

, Dominiv

, Feeney

. (1997) Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Advanced Drug Delivery Reviews, 23, 3-25.

13.

Lipinski

, Lombardo

, Dominiv

, Feeney

. (2001) Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Advanced Drug Delivery Reviews, 46, 3-26.

14.

Veber

, Johnson

, Cheng

H-Y

, Smith

, Ward

, Kopple

. (2002) Molecular properties that influence the oral bioavailability of drug candidates. Journal of Medicinal Chemistry, 45, 2615-2623.

15.

Murray

RDH

, Méndez

, Brown

. (1982) Identification and structural elucidation. In The Natural Coumarins. John Wiley & Sons Ltd, Bristol, England, 22–38.

16.

Ito

, Furukawa

(1987) Constituents of Murraya exotica L. Structure elucidation of new coumarins. Chemical and Pharmaceutical Bulletin, 35, 4277-4285.

17.

Imai

, Kinoshita

, Sankawa

(1989) Constituents of leaves of Murraya paniculata collected in Taiwan. Chemical and Pharmaceutical Bulletin, 37, 358-362.

18.

Yan

, Shen

, Liu

, Zou

, Xu

(2018) Preparative isolation and purification of hainanmurpanin, meranzin, and phebalosin from leaves of Murraya exotica L. using supercritical fluid extraction combined with consecutive high-speed countercurrent chromatography. Journal of Separation Science, 41, 2092-2101.

19.

Joshi

, Kamat

. (1970) Isolation of 3,3',4',5,5',7,8-heptamethoxyflavone from Murraya exotica. Phytochemistry, 9, 889.

20.

Dreyer

. (1968) Chemotaxonomy of the Rutaceae. IV. Constituents of Murraya paniculata (Linn.) Jack. Journal of Organic Chemistry, 33, 3574-3576.

21.

Ito

, Fujiwara

, Koizumi

, Furukawa

(1998) Isolation and characterization of an antibacterial substance from Citrus plant. Journal of the Chinese Chemical Society, 45, 89-91.

22.

Machida

, Osawa

(1989) On the flavonoid constituents from the peels of Citrus hassaku Hort. ex Tanaka. Chemical and Pharmaceutical Bulletin, 37, 1092-1094.

23.

Joshi

, Kamat

. (1969) Structure of exoticin, a flavone from the leaves of Murraya exotica Linn. Indian Journal of Chemistry, 7, 636.

24.

Ramstad

, Lin

W-NC.

(1968) Coumurrayin, a new coumarin from Murraya paniculata (L.) Jack. Tetrahedron Letters, 7, 811-813.

25.

Singhuber

, Baburin

, Ecker

, Kopp

, Hering

(2011) Insights into structure-activity relationship of GABA_A receptor modulating coumarins and furanocoumarins. European Journal of Pharmacology, 668, 57-68.

26.

Hanrahan

, Chebib

, Johnston

GAR

. (2015) Interactions of flavonoids with ionotropic GABA receptors. Advances in Pharmacology, 72, 189-200.

27.

Wasowski

, Marder

(2012) Flavonoids as GABA_A receptor ligands: the whole story? Journal of Experimental Pharmacology, 4, 9-24.

28.

Rueda

, De Mieri

, Hering

, Hamburger

(2014) HPLC-based activity profiling for GABA_A receptor modulators in Adenocarpus cincinnatus. Journal of Natural Products, 96, 640-649.

29.

Westerfield

(2007) A Guide for the laboratory use of zebrafish (Danio rerio). University of Oregon Press, Eugene, USA.

30.

Potterat

, Hamburger

(2013) Concepts and technologies for tracking bioactive compounds in natural product extracts: generation of libraries, and hyphenation of analytical processes with bioassay. Natural Product Reports, 30, 546-564.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.20 MB