SPECTRAL CHARACTERISTICS OF CYTOCHROME P45O IN THE INTERACTION WITH PROPOXAZEPAM AND ITS METABOLITE
DOI:
https://doi.org/10.11603/mcch.2410-681X.2023.i2.13854Keywords:
Propoxazepam, CYP, rat liver microsomes, spectral changesAbstract
Introduction. Propoxazepam (a 1,4-benzodiazepine derivative) is a novel analgesic that simultaneously inhibits acute and chronic pain with anti-inflammatory and anticonvulsant components that depend on the state of the gamma-aminobutyric acid receptor. There is currently no information on the possible interaction of the compound with other drugs at the cytochrome P-450 (CYP) level.
The aim of the study – to study the indicators of spectral changes of total CYP of rat liver microsomes when interacting with propoxazepam and its 3-hydroxymetabolite, which may indirectly indicate the interaction of the compound with other drugs.
Research Methods. Experiments were conducted on mature Wistar rats weighing 150–180 g, which were divided into two groups: 1 – animals that were treated with phenobarbital (80 mg/kg) for 3 days); 2 – which was treated with 3-methylcholanthrene (40 mg/kg) for 3 days. Rat liver microsomes were isolated by ultracentrifugation of the postmitochondrial fraction in a medium containing 1.15 % KCl and 40 mM Tris-HCl buffer pH 7.4 in a Beckman ultracentrifuge at 105,000 g. Microsomes were diluted with 40 mM Tris-HCl buffer pH 7.4 to a protein concentration of 2.6 mg/ml. Propoxazepam and its 3-hydroxymetabolite were dissolved in methanol to a concentration of 30 mM and added to 3 ml of microsome suspension. The content of CYP and its spectral characteristics were studied by the method of differential spectrophotometry on the “Aminco” spectrophotometer.
Results and Discussion. Propoxazepam (I) and 3-hydroxymetabolite (II) interacting with CYP of the liver of rats administered phenobarbital and 3-methylcholantren demonstrated the 2nd type of spectral changes in hemoprotein. The binding constans of I and II considerably differ, which points to the possibility of interaction of substrates with different areas of CYP. The characteristic points (ma[imum, minimum and isobestic) of CYP spectrum were displaced during II titration to the longwawe region as compared with I. Propoxazepam and its metabolite showed negligible affinity to microsomes of rats treated with different inductors, Ks values were in the range 0.76·10-4 – 1.25·10-4 М.
Conclusions. Quantitative indicators of the inhibitory activities of I and II using the method of differential spectroscopy are only indicative in nature, but still can suggest at least a significant possibility of inhibitory interaction in clinical use of drugs.
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