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Home » Archive » 2021 » Veterinary Session

Veterinary session

In vitro comparison of biotransformation of 3-amidinophenylalanine-type compounds in different animal species
Fedor Zsófia - year 5
University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology
Supervisor: Pásztiné Dr. Erzsébet Gere

Abstract:

The blocking effect of 3-amidinophenylalanine-derived (3AphA) compounds on transmembrane protease serine 2 (TMPRSS2) makes these substances important drug candidates. TMPRSS2 activity has a key role in the development and negative prognosis of different epithelial-derived tumors (breast and prostate cancer) and in the entry into the cells and replications of some viruses (coronavirus, influenzavirus, parainfluenzavirus). Due to these facts, more extensive study including in vitro pharmacotoxicological analysis of these compounds is required from therapeutical application point of view.

The characterize the interaction between liver and 3APhA-derived matriptase/TMPRSS2 (MI) inhibitors in vitro changes in cell viability, alterations in levels of oxidative stress and in cytochrome P450 (CYP) isoenzymes’ activities were monitored. Rat, beagle and cynomolgus monkey hepatocytes were treated with different concentrations (10, 20, 50 and 100 µM) of matriptase/TMPRSS2 inhibitors such as MI-1900 and MI-432. Cell viability was measured by MTS assay, extracellular hydrogen peroxide levels were detected by Amplex Red method, CYP enzyme activity was analyzed by luminescence technique and occludin localization was determined by confocal microscopy using immunofluorescence staining.

Our results showed that none of the tested MI-compounds affected the viability of cells from the tested animal species in 100 µM concentration. After the 4-hour treatment the rat hepatocytes did not show significant changes in extracellular hydrogen peroxide production. In beagle hepatocytes, MI-1900 at 50 µM decreased the hydrogen peroxide levels significantly after 4 hours, which was not detectable after 24 hours. The treatment with 50 µM MI-432 for 4 hours resulted in a significant reduction in cynomolgus monkey hepatocytes, while treatment with 10 and 20 µM MI-1900 led to significant increase in oxidative stress. After 24-hour treatment, MI-1900 at 50 µM significantly reduced hydrogen peroxide levels in both rat and monkey hepatocytes.

The activity of the CYP1A2 in rats was not significantly affected by any concentration of the applied inhibitors. In beagle hepatocytes a substance-dependent (MI-432, 20 and 50 µM) inhibition, in monkey hepatocytes a concentration-dependent blockage (MI-432 and MI-1900, 50 µM) could be observed.

In conclusion, due to the inhibitory effects of 3AphA derivatives on activities of the CYP1A2 isoenzyme it is recommended to monitor drug interactions closely in their future therapeutical application in the treatment of viral diseases such as SARS-CoV-2.



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