|Title||The development of improved and new in vitro assays for detecting the genotoxic and non-genotoxic carcinogenic potential of chemicals in the discovery phase of drug development|
|Source||University. Promotor(en): Ivonne Rietjens; John Groten, co-promotor(en): W.G.E.J. Schoonen. - [S.l. : S.n. - ISBN 9789085858478 - 288|
Sub-department of Toxicology
|Publication type||Dissertation, internally prepared|
|Keyword(s)||genotoxiciteit - carcinogenese - nieuwe geneesmiddelen - geneesmiddelenontwikkeling - in vitro - in vitro kweek - genotoxicity - carcinogenesis - new drugs - drug development - in vitro culture|
|Categories||Carcinogenesis, Mutagenesis, Teratology, Reproduction Toxicity|
|Abstract||In drug development, toxicity is an important factor for attrition, resulting in a failure rate of 30%-40%. Hepatotoxicity, nephrotoxicity, cardiovascular safety, reproduction toxicity, developmental toxicity (teratogenicity), genotoxicity and carcinogenicity are the main causes for attrition in safety assessment.
Screening on these aspects in the early discovery phase of drug development and using these data for compound optimization and deselection might result in drug development candidates with an improved success rate. The present thesis focused on early screening for genotoxicity and carcinogenicity.
In recent years some progress has been made with assays to assess genotoxicity and non-genotoxic carcinogenicity at the end of the discovery phase. However, the time point at which these genotoxicity and carcinogenicity assays are performed is still relatively late, only a few assays for such a strategy are available, the throughput of these assays is in general still low and most of them have not yet been validated extensively. An additional drawback is that the currently used in vitro assays for the detection of genotoxicity give a high rate of false positive results, which makes application in the early discovery phase of drug development cumbersome.
The goal of the present thesis was therefore to develop improved and new in vitro assays for detecting the genotoxic and non-genotoxic carcinogenic potential of chemicals, validate these assays with proper reference compounds, and to develop a strategy for application of these assays in the early discovery phase of drug development.
High throughput assays based on bacteria, yeast and human/rodent cell lines were developed. In the case of human cell lines, the focus was on the HepG2 cell line as the properties of HepG2 cells are expected to give a good prediction for in vivo genotoxicity. The results in this thesis show that an early prediction can be made for bacterial mutagenicity (gene mutations), mammalian genotoxicity (chromosome damage), and non-genotoxic carcinogenic potential by aryl hydrocarbon receptor (AhR) activation. To develop a strategy for application of the HTS genotoxicity assays in the early discovery phase several combinations of assays were evaluated. The combination VitotoxTM + HepG2 p53 reporter assay was based on the presented results in this thesis the most useful for screening compounds for their genotoxic potential in the early drug discovery phase without the risk on high numbers of false positives. CYP1A induction assays in human HepG2 and rat H4IIE cells may be performed in parallel with these assays to be able to detect non-genotoxic carcinogenic potential by AhR activators. Further application of these assays may prove useful in future drug development strategies.