Ukr.Biochem.J. 2014; Volume 86, Issue 4, Jul-Aug, pp. 150-157


Kinetics of hydrolysis of 1,4-benzodiazepine derivative by carboxylesterases in mice organism

M. Ya. Golovenko, V. B. Larionov

A. V. Bogatsky Physics-Chemical Institute, National Academy of Sciences of Ukraine, Odesa;

Chemical modification of the physiologically active substances and creation of prodrugs is one of the ways for pharmacotherapy optimization. The aim of the work was determination of the kinetic parameters of nonspecific esterases that catalyze hydrolysis of new hypnotic drug Levana (1,4-benzodiazepine derivative). The experiments were carried out using the 14C-labelled Levana and its active metabolite – 3-hydrixyphenazepam. In vitro it was shown that Levana undergoes spontaneous hydrolysis even in buffer solution (pH 7.4), though in plasma and homogenates of brain and liver this process is more intensive (conventional Vmax was 6.9 ± 0.5, 19 ± 4 and 12 ± 1 mM/(h∙mg of protein, correspondingly). The samples mentioned differ by activity of tissue estera­ses being most active in the liver (conventional Km 0.45 ± 0.04 mM for the liver and 47 ± 11 mM for the brain). In plasma carboxylesterase activity (for Leva­na) is the lowest (conventional Km 129 ± 10 mM). In vivo it was shown that Levana more easily permeates brain-blood barrier (compared to 3-hydroxyphenaze­pam), that leads to higher concentrations (after hyd­rolysis) of its metabolite in brain tissue. Also it is quantitatively estimated as the increase of concentration (brain/blood) ratio  ~1.4 times.

Keywords: , , , ,


  1. Andronati SA, Avrutskiy GYa, Bogats­kiy AB, Voronina TA. et al. Phenazepam. K.: Nauk. Dumka, 2003.  219 p. (In Russian).
  2. Andronati SA, Karaseva TL, Popova LV, Makan SYu, Boyko IA, Bitenskiy BS. GABA-ergic hypnotic drugs. Vistn. Psychiatr. Psychopharmacother. 2004;5(1):6–17. (In Russian).
  3. Golovenko NYa, Zinkovskiy VG, Yacubov­skaya LN. Synthesis of 1.4-benzodiazepine derivatives, labeled with radioactive isotopes, and determination of their metabolite’s structure. Ukr Chem J. 1999;65(9):34–44. (In Russian).
  4. Golovenko NYa, Zinkovskiy VG. Determination of benzodiazepine tranquilizers and their metabolites in biological samples. Chem. Pharm. J. 1978;12(1):1–11. (In Russian). CrossRef
  5. Keleti Т. Basic enzyme kinetics. M.: Mir, 1990. 348 p. (In Russian).
  6. Ono S, Hatanaka T, Miyazawa S, Tsutsui M, Aoyama T, Gonzalez FJ, Satoh T. Human liver microsomal diazepam metabolism using cDNA-expressed cytochrome P450s: role of CYP2B6, 2C19 and the 3A subfamily. Xenobiotica. 1996 Nov;26(11):1155-66. PubMed, CrossRef
  7. Golovenko MYa, Maltsev EV, Larionov VB. Kinetics of the chemical hydrolysis of hypnotic drug “Levana IC”. Pharmaceut. J. 2010;(4):79–87. (In Ukrainian).
  8. Golovenko NYa. Physics-Chemical Pharma­cology. Odessa: Astroprint, 2004. 720 p. (In Russian).
  9. Golovenko NYa, Kravchenko IA. Bioche­mical pharmacology of prodrugs. Odessa: “Ecologiya”, 2007. 360 p. (In Russian).
  10. Satoh T, Hosokawa M. The mammalian carboxylesterases: from molecules to functions. Annu Rev Pharmacol Toxicol. 1998;38(1):257-88. Review. PubMed, CrossRef
  11. Andronati SA, Shesterenko EA, Sevastya­nov OB, Romanovskaya II, Pavlovskiy VI, Semenishina KO, Osetrov VE. Isolation and characterization of carboxylesterase from piggy liver and its usage in stereoselective hydrolysis of 1.4-benzodiazepine-2one derivatives. Biotechnology. 2011;4(5):71–76. (In Ukrainian).
  12. Redinbo MR, Bencharit S, Potter PM. Human carboxylesterase 1: from drug metabolism to drug discovery. Biochem Soc Trans. 2008 Jun;31(Pt 3):620-4. PubMed, CrossRef
  13. Zhang J, Burnell JC, Dumaual N, Bosron WF. Binding and hydrolysis of meperidine by human liver carboxylesterase hCE-1. J Pharmacol Exp Ther. 1999 Jul;290(1):314-8. PubMed
  14. Xie M, Yang D, Wu M, Xue B, Yan B. Mouse liver and kidney carboxylesterase (M-LK) rapidly hydrolyzes antitumor prodrug irinotecan and the N-terminal three quarter sequence determines substrate selectivity. Drug Metab Dispos. 2003 Jan;31(1):21-7. PubMed, CrossRef

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License.