Identification Through High-throughput Screening of 4'-methoxyflavone and 3',4'-dimethoxyflavone As Novel Neuroprotective Inhibitors of Parthanatos

Overview

Journal Br J Pharmacol

Publisher Wiley

Specialty Pharmacology

Date 2013 Apr 5

PMID 23550801

Citations 18

Authors

A A Fatokun

J O Liu

V L Dawson

T M Dawson

Affiliations

Soon will be listed here.

Abstract

Background And Purpose: The current lack of disease-modifying therapeutics to manage neurological and neurodegenerative conditions justifies the development of more efficacious agents. One distinct pathway leading to neuronal death in these conditions and which represents a very promising and attractive therapeutic target is parthanatos, involving overactivation of PARP-1. We therefore sought to identify small molecules that could be neuroprotective by targeting the pathway.

Experimental Approach: Using HeLa cells, we developed and optimized an assay for high-throughput screening of about 5120 small molecules. Structure-activity relationship (SAR) study was carried out in HeLa and SH-SY5Y cells for molecules related to the initial active compound. The neuroprotective ability of each active compound was tested in cortical neuronal cultures.

Key Results: 4'-Methoxyflavone (4MF) showed activity by preventing the decrease in cell viability of HeLa and SH-SY5Y cells caused by the DNA-alkylating agent, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), which induces parthanatos. A similar compound from the SAR study, 3',4'-dimethoxyflavone (DMF), also showed significant activity. Both compounds reduced the synthesis and accumulation of poly (ADP-ribose) polymer and protected cortical neurones against cell death induced by NMDA.

Conclusions And Implications: Our data reveal additional neuroprotective members of the flavone class of flavonoids and show that methoxylation of the parent flavone structure at position 4' confers parthanatos-inhibiting activity while additional methoxylation at position 3', reported by others to improve metabolic stability, does not destroy the activity. These molecules may therefore serve as leads for the development of novel neurotherapeutics for the management of neurological and neurodegenerative conditions.

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References

Zhang X, Chen J, Graham S, Du L, Kochanek P, Draviam R . Intranuclear localization of apoptosis-inducing factor (AIF) and large scale DNA fragmentation after traumatic brain injury in rats and in neuronal cultures exposed to peroxynitrite. J Neurochem. 2002; 82(1):181-91. DOI: 10.1046/j.1471-4159.2002.00975.x. View

Dugas Jr A, Bonin G, Price K, Fischer N, Winston G . Evaluation of the total peroxyl radical-scavenging capacity of flavonoids: structure-activity relationships. J Nat Prod. 2000; 63(3):327-31. DOI: 10.1021/np990352n. View

Lin J, Liang Y, Lin-Shiau S . Cancer chemoprevention by tea polyphenols through mitotic signal transduction blockade. Biochem Pharmacol. 1999; 58(6):911-5. DOI: 10.1016/s0006-2952(99)00112-4. View

Yashiroda Y, Okamoto R, Hatsugai K, Takemoto Y, Goshima N, Saito T . A novel yeast cell-based screen identifies flavone as a tankyrase inhibitor. Biochem Biophys Res Commun. 2010; 394(3):569-73. DOI: 10.1016/j.bbrc.2010.03.021. View

MIDDLETON Jr E, Kandaswami C, Theoharides T . The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer. Pharmacol Rev. 2000; 52(4):673-751. View

Li X, Nemoto M, Xu Z, Yu S, Shimoji M, Andrabi S . Influence of duration of focal cerebral ischemia and neuronal nitric oxide synthase on translocation of apoptosis-inducing factor to the nucleus. Neuroscience. 2006; 144(1):56-65. PMC: 1876769. DOI: 10.1016/j.neuroscience.2006.08.065. View

SCHROETER H, Spencer J, Williams R . Flavonoids protect neurons from oxidized low-density-lipoprotein-induced apoptosis involving c-Jun N-terminal kinase (JNK), c-Jun and caspase-3. Biochem J. 2001; 358(Pt 3):547-57. PMC: 1222091. DOI: 10.1042/0264-6021:3580547. View

Chong C, Xu J, Lu J, Bhat S, Sullivan Jr D, Liu J . Inhibition of angiogenesis by the antifungal drug itraconazole. ACS Chem Biol. 2007; 2(4):263-70. DOI: 10.1021/cb600362d. View

Takano K, Tabata Y, Kitao Y, Murakami R, Suzuki H, Yamada M . Methoxyflavones protect cells against endoplasmic reticulum stress and neurotoxin. Am J Physiol Cell Physiol. 2006; 292(1):C353-61. DOI: 10.1152/ajpcell.00388.2006. View

10.

Nagase H, Omae N, Omori A, Nakagawasai O, Tadano T, Yokosuka A . Nobiletin and its related flavonoids with CRE-dependent transcription-stimulating and neuritegenic activities. Biochem Biophys Res Commun. 2005; 337(4):1330-6. DOI: 10.1016/j.bbrc.2005.10.001. View

11.

Zhang , Chung , OLDENBURG . A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. J Biomol Screen. 2000; 4(2):67-73. DOI: 10.1177/108705719900400206. View

12.

Fernandez S, Karim N, Mewett K, Chebib M, Johnston G, Hanrahan J . Flavan-3-ol esters: new agents for exploring modulatory sites on GABA(A) receptors. Br J Pharmacol. 2011; 165(4):965-77. PMC: 3312492. DOI: 10.1111/j.1476-5381.2011.01615.x. View

13.

Hanneken A, Lin F, Johnson J, Maher P . Flavonoids protect human retinal pigment epithelial cells from oxidative-stress-induced death. Invest Ophthalmol Vis Sci. 2006; 47(7):3164-77. DOI: 10.1167/iovs.04-1369. View

14.

Arora A, Nair M, Strasburg G . Structure-activity relationships for antioxidant activities of a series of flavonoids in a liposomal system. Free Radic Biol Med. 1998; 24(9):1355-63. DOI: 10.1016/s0891-5849(97)00458-9. View

15.

Nilsson J, Nielsen E, Liljefors T, Nielsen M, Sterner O . Azaflavones compared to flavones as ligands to the benzodiazepine binding site of brain GABA(A) receptors. Bioorg Med Chem Lett. 2008; 18(21):5713-6. DOI: 10.1016/j.bmcl.2008.09.092. View

16.

Andrabi S, Dawson T, Dawson V . Mitochondrial and nuclear cross talk in cell death: parthanatos. Ann N Y Acad Sci. 2008; 1147:233-41. PMC: 4454457. DOI: 10.1196/annals.1427.014. View

17.

Sykes M, Avery V . Development of an Alamar Blue viability assay in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427. Am J Trop Med Hyg. 2009; 81(4):665-74. DOI: 10.4269/ajtmh.2009.09-0015. View

18.

Walle U, Walle T . Bioavailable flavonoids: cytochrome P450-mediated metabolism of methoxyflavones. Drug Metab Dispos. 2007; 35(11):1985-9. DOI: 10.1124/dmd.107.016782. View

19.

Dawson V, Dawson T, Bartley D, Uhl G, Snyder S . Mechanisms of nitric oxide-mediated neurotoxicity in primary brain cultures. J Neurosci. 1993; 13(6):2651-61. PMC: 6576487. View

20.

Andrabi S, Kim N, Yu S, Wang H, Koh D, Sasaki M . Poly(ADP-ribose) (PAR) polymer is a death signal. Proc Natl Acad Sci U S A. 2006; 103(48):18308-13. PMC: 1838747. DOI: 10.1073/pnas.0606526103. View