Pharmacology and Toxicology Unit

Responsible person: Assoc. Prof. PharmDr. Ondřej Zendulka, Ph.D.

The unit will provide space for basic pharmacological and toxicological research on substances in in vivo rat and mouse models. It will primarily serve for the preparation of application solutions of substances, premedication of animals, and collection of biological samples for subsequent determination of plasma and tissue levels of substances and their metabolites, histological and biochemical determinations.

The unit's equipment will include the necessary instrumentation for performing all application methods, including intravenous administration using implanted catheters and intracerebral stereotactic injections, as well as the equipment necessary for performing in vivo pharmacokinetic studies (metabolic cages, devices for repeated blood sampling from freely moving animals) and basic sample processing (centrifuge).

It is expected that animal dissection and basic toxicological screening, including histopathology and haematotoxicology, will be performed in collaboration with the Histopathology and haematology Unit. Cardiological toxicity, or the effect of the tested substances on myocardial ion channels, will be determined in collaboration with the electrophysiology laboratory and the patch clamp laboratory, respectively.

What services/products will the unit provide?

Cytochrome P450 inhibition study with RLM

pilot experiment + determination of IC50 + Ki (interaction between the tested substance and CYP (1A, 2C, 2D, 3A) at the level of rat liver microsomal fraction using the method of monitoring the rate of metabolism of selective substrates)

Cytochrome P450 inhibition study with HLM

pilot experiment + determination of IC50 + Ki (interaction between the test substance and CYP (1A2, 2C9, 2D6, 3A4) at the level of human liver microsomal fraction by monitoring the metabolism rate of selective substrates)

Isolation of hepatic microsomal fraction

from supplied tissue + characterization (protein content, CYP content)

In vivo experiment with premedication of animals

effect on CYP metabolic activity (1A, 2C, 2D, 3A) (determination of the effect of the test substance on CYP enzymes in vivo (laboratory rat) by monitoring the metabolism rate of selective substrates)

Determination of basic pharmacokinetic parameters in in vivo model

development of an analytical method for the detection of the tested substance and its metabolites in biological matrices + determination of basic pharmacokinetic parameters (T_max, V_d, K_e, t_1/2, CL) after single and repeated administration in mice/rats, including determination of the main route of elimination using metabolic cages

Development and validation of methods for determining low molecular weight substances (Mw < 1000) in biological matrices

complete development and validation of custom methods, including extraction procedures. Methods: UHPLC+DAD, FLD, MS, MS/MS; LLE, SPE

Development of a chromatographic method for the separation/purification/isolation of substances from mixtures/Flash/preparative chromatography (LC)

isolation and purification from mixtures using flash/preparative chromatography (small organic molecules, natural products, metabolite isolation, trace enrichment)

Microwave synthesis

development of a method for synthesizing the final product (microwave-assisted synthesis of a wide range of organic compounds, including heterocyclic and peptide syntheses, polymerizations, and organometallic reactions)

Detection of the metabolism of the tested substance by cytochrome P450

development of an analytical method for detecting the test substance and its metabolites in biological matrices + verification of whether the test substance is a substrate of human and rat CYP, with the possibility of identifying the metabolites produced and the specific CYP involved in biotransformation

Support in planning the design of experiments

administration related to the handling of the experimental project
optimization of the route of administration of the test substance (intragastric, s.c., i.m., i.v., intracerebral, intracerebroventricular, etc.)
solubility tests and optimization of the solvent for the test substance for the in vivo model
proposal of the dose of the administered substance and sampling intervals

(please consult specific details in advance with Assoc.Prof. Zendulka, email: zendulka@med.muni.cz)