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DARU Journal of Pharmaceutical Sciences

, Volume 26, Issue 2, pp 191–198 | Cite as

New mechanisms of phenytoin in calcium homeostasis: competitive inhibition of CD38 in hippocampal cells

  • Leila SadeghiEmail author
  • Reza Yekta
  • Gholamreza Dehghan
Research Article
  • 156 Downloads

Abstract

Purpose

Phenytoin is a major anticonvulsant drug that is effective to improve arrhythmia and neuropathic pain. According to early works, phenytoin affected cell membrane depolarization by sodium channel blocking, guanylyl and adenylyl cyclase suppression that cause to intracellular Na+ and Ca2+ downregulation. This study was aimed to clarify some ambiguities in pathophysiological action of phenytoin by in vitro and molecular docking analyses.

Methods

In this study intracellular free Ca2+ of primary culture of embryonic mouse hippocampus evaluated via Fura 2 as fluorescent probe. The effects of phenytoin on ADP ribosyl cyclase activity was assessed by recently developed fluorometric assay. Molecular docking simulation was also implemented to investigate the possible interaction between phenytoin and CD38.

Results

Our results confirmed phenytoin competitively inhibits cyclase activity of CD38 (IC50 = 8.1 μM) and reduces cADPR content. cADPR is a Ca2+-mobilising second messenger which binds to L-type calcium channel and ryanodine receptors in cell and ER membrane and increases cytosolic free Ca2+. Ca2+ content of cells decreased significantly in the presence of phenytoin in a dose dependent manner (IC50 = 12.74 µM). Based on molecular docking analysis, phenytoin binds to deeper site of CD38 active site, mainly via hydrophobic interactions and consequently inhibits proper contact of substrate with catalytic residues specially Glu 226, Trp 186, Thr221.

Conclusion

Taken together, one of the anticonvulsant mechanisms of phenytoin is Ca2+ inhibition from CD38 pathway, therefore could be used in disorders that accompanied by CD38 over production or activation such as heart disease, depression, brain sepsis, airway disease, oxidative stress and inflammation.

Graphical abstract

Keywords

Phenytoin CD38 Competitive inhibition Calcium homeostasis Sodium blocker Membrane depolarization 

Notes

Compliance with ethical standards

Conflict of interest

All of the Authors have no conflict of interest to declare.

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Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Department of Animal Biology, Faculty of Natural SciencesUniversity of TabrizTabrizIran

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