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| Departments and Laboratories: |
 
 
The Juliusz Slowacki Theatre (Cracow)
The Barbicane Fortification (Cracow)
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Department of Pharmacokinetics and Drug Metabolism
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| Scope of Research
| Methods
| Cooperation
| Research Staff |
Head: Prof. Władysława Anna Daniel
Phone: (+48 12) 66 23 266
E-mail: nfdaniel@cyf-kr.edu.pl
Scope of Research
- contribution of brain cytochromes P450 to the local metabolism of monoaminergic neurotransmitters and psychoactive drugs;
- role of the central nervous system in the regulation of cytochrome P450 expression;
- interaction between psychotropic drugs and cytochrome P450 isoenzymes in the brain and liver;
- identification and quantitative contribution of liver cytochrome P450 isoenzymes to the metabolism of centrally active drugs;
- pharmacokinetic interactions within clinical combinations of psychotropics;
- mechanisms of pharmacokinetic interactions;
- pharmacokinetics and metabolism of psychotropic drugs after acute and chronic treatment;
- relationships between drug concentration in blood plasma and the brain: mechanisms of drug distribution.
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Methods
- Pharmacokinetics and drug metabolism:
- determination of the concentrations of psychotropic drugs and their metabolites in blood plasma, brain and peripheral tissues using HPLC
(UV and fluorescence detection);
- drug metabolism in vitro: measurement of the rate of drug metabolism in liver microsomes in the absence or presence of cytochrome
P450 specific inhibitors - kinetic study, calculation of Km, Vmax, CL and the inhibition constant Ki (HPLC);
- assessment of the ability of cDNA-expressed cytochrome P450 isoenzymes to catalyse metabolic reactions - calculation of kinetic parameters,
Km, Vmax and CL (HPLC);
- distribution study: assessment of tissue drug uptake in vitro (HPLC, spectrofluorimetry or liquid scintillation counting).
- Biochemistry:
- determination of the concentrations of monoaminergic neurotransmitters and their metabolites in the brain using HPLC (electrochemical detection);
- estimation of cytochrome P450 activity in brain and liver microsomes by isoenzyme-specific reactions (HPLC);
- assessment of the concentrations of cytochrome P450 specific substrates (testosterone, caffeine, ethylmorphine, bufuralol, warfarin)
and their metabolites in brain and liver microsomes using HPLC (UV and fluorescence detection);
- evaluation of total cytochrome P450 and cytochrome b-5 concentrations in liver microsomes (double-beam spectrophotometry;
- assessment of the ability of liver/brain microsomes and cDNA-expressed cytochrome P450 isoenzymes to catalyze endogenous substrate metabolism
- calculation of kinetic parameters, Km, Vmax and CL, Ki (HPLC);
- determination of the concentrations of cytochrome P450 isoenzyme proteins in brain and liver microsomes by electrophoresis and Western blot,
followed by immunodetection and chemoluminescence measurement (Luminescent Image Analyzer LAS-100);
- determination of hormone and cytokine levels in blood plasma/serum (ELISA, Multiscan Spectrum UV microplate reader).
- Others:
- stereotaxic implantation of cannula in brain structures (intracerebral injections).
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International Cooperation
The Department cooperates with:
- the Institut National de la Santé et de la Rechere Medicale (INSERM), Montpellier (France)
- - cytochrome P450 expression in steam cells and hepatocytes.
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Research Staff:
Engineers, Technicians, and Other Employees:
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