Screening of NOS activity and selectivity of newly synthesized acetamidines using RP-HPLC

J Pharm Biomed Anal. 2016 Feb 20:120:419-24. doi: 10.1016/j.jpba.2015.11.045. Epub 2015 Dec 2.

Abstract

Nitric Oxide Synthase (NOS) inhibitors could play a powerful role in inflammatory and neurodegenerative diseases. In this work, novel acetamidine derivatives of NOS were synthesized and the inhibitor activity was evalued. To screen the activity and selectivity, the l-citrulline residue, after the enzymatic NOS assay, was derivatized with o-phthaldialdehyde/N-acetyl cysteine (OPA/NAC) and then evaluated by RP-HPLC method with fluorescence detection. All compounds did not affect the activity of endothelial and neuronal isoforms, while nine of them possessed a percentage of iNOS activity at 10μM lower than 50%, and were selected for IC50 evaluation. Among them, a compound emerged as a very potent (IC50 of 53nM) and selective iNOS inhibitor.

Keywords: Acetamidine; Fluorescence detection; High-performance liquid chromatography; Nitric Oxide Synthase inhibitor; OPA/NAC derivatization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidines / chemical synthesis*
  • Amidines / pharmacokinetics
  • Chromatography, High Pressure Liquid / methods
  • Enzyme Activation / drug effects
  • Enzyme Activation / physiology
  • Enzyme Inhibitors / analysis*
  • Enzyme Inhibitors / pharmacokinetics
  • Nitric Oxide Synthase Type II / analysis*
  • Nitric Oxide Synthase Type II / antagonists & inhibitors*
  • Nitric Oxide Synthase Type II / metabolism

Substances

  • Amidines
  • Enzyme Inhibitors
  • Nitric Oxide Synthase Type II
  • acetamidine