title: Characterization of inhibitory effects of the potential therapeutic inhibitors, benzoic acid and pyridine derivatives, on the monophenolase and diphenolase activities of tyrosinase.
abstract: OBJECTIVE
  Involvement of tyrosinase in the synthesis of melanin and cell signaling pathway has made it an attractive target in the search for therapeutic inhibitors for treatment of different skin hyperpigmentation disorders and melanoma cancers.
  
  
  METHODS
  In the present study, we conducted a comprehensive kinetic analysis to understand the mechanisms of inhibition imposed by 2-amino benzoic acid, 4-amino benzoic acid, nicotinic acid, and picolinic acid on the monophenolase and diphenolase activities of the mushroom tyrosinase, and then MTT assay was exploited to evaluate their toxicity on the melanoma cells.
  
  
  RESULTS
  Kinetic analysis revealed that nicotinic acid and picolinic acid competitively restricted the monophenolase activity with inhibition constants (Ki) of 1.21 mM and 1.97 mM and the diphenolase activity with Kis of 2.4 mM and 2.93 mM, respectively. 2-aminobenzoic acid and 4-aminobenzoic acid inhibited the monophenolase activity in a non-competitive fashion with Kis of 5.15 µM and 3.8 µM and the diphenolase activity with Kis of 4.72 µM and 20 µM, respectively.
  
  
  CONCLUSIONS
  Our cell-based data revealed that only the pyridine derivatives imposed cytotoxicity in melanoma cells. Importantly, the concentrations of the inhibitors leading to 50% decrease in the cell density (IC50) were comparable to those causing 50% drop in the enzyme activity, implying that the observed cytotoxicity is highly likely due to the tyrosinase inhibition. Moreover, our cell-based data exhibited that the pyridine derivatives acted as anti-proliferative agents, perhaps inducing cytotoxicity in the melanoma cells through inhibition of the tyrosinase activities.
authors: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors: Nematollah
authors: N
authors: Gheibi
authors: Department of Biology, Faculty of Basic Sciences, Islamic Azad University of Science and Research, Tehran, Iran.
authors: Negar
authors: N
authors: Taherkhani
authors: Department of Biology, Faculty of Basic Sciences, Islamic Azad University of Science and Research, Tehran, Iran.
authors: Abolfazl
authors: A
authors: Ahmadi
authors: Biochemistry and Biophysics Department, National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
authors: Kamahldin
authors: K
authors: Haghbeen
authors: Department of Clinical Biochemistry and Genetics, Qazvin University of Medical Science, Qazvin, Iran.
authors: Dariush
authors: D
authors: Ilghari
keywords: 2-amino benzoic acid; 4-amino benzoic acid; Diphenolase activity; Inhibition; Monophenolase activity; Mushroom tyrosinase; Nicotinic acid; Picolinic acid