Naslov
Cell-based in vitro toxicology screening for the macrolide class of compounds

Autori
Jelić, Dubravko ; Čulić, Ognjen ; Verbanac, Donatella ; Brajša, Karmen ; Perović, Daniela ; Mildner, Boris ; Bokulić, Ana ; Radošević, Dražen ; Polančec, Denis ; Tomašković, Marija ; Antolović, Roberto

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, stručni

Izvornik
World Pharmaceutical Congress 2005 : Abstract book / - Philadelphia (PA) : Cambridge Healthtech Institute (CHI), 2005, P 20-P 20

Skup
World Pharmaceutical Congress 2005

Mjesto i datum
Philadelphia (PA), Sjedinjene Američke Države, 24.05.2005. - 26.05.2005

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
screening; in vitro toxicology; macrolides

Sažetak
High-throughput prediction of potential drug toxicity is necessary in early stage of drug discovery while testing potential hits. This is also a part of the lead optimization process and one of major focus in discovery phase of drug development. In this context, setting-up of an in vitro screening system reflecting possible in vivo toxicity is pre-requisite for the assessment of an early safety profile of the new chemical entity. Cell viability assays are very popular and broadly used. Since it is hard to say which of the assays available so far is the most appropriate, in our Institute we introduced several defferent methods to determine cell cytotoxicity/viability in a high-throughput format. We are focused on macrolide compounds, a promising compounds class that target different types of anti-infective and/or anti-inflammatory targets. Our experience with classic cell viability or cytotoxicity assays such as tetrazolium-based colorimetric assays using MTT or MTS, assays that detect lactate dehydrogenase (LDH), bioluminiscent ATP cytotoxicity and [3H] thymidine incorporation assays on macrolide class of compounds will be described. In addition, using flow cytometry and high-speed cell sorting technology, different cell types of human, mouse or rat origin can be isolated for cytotoxicity assays on specific subpopulations. There are very diverse markers that indicate the number of dead cells in cytotoxicity assays, number of live cells in viability assays, total cell numbers, or specific mechanism of cell death (apoptosis or necrosis). Detection methods are mostly based on the cell membrane integrity and cell metabolic activity measuring ATP or NADH cell production. Our conclusion is that it is important to choose appropriate end-point measurement, based on the compound class to be tested.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

POVEZANOST RADA