Comparative quantification of pharmacodynamic parameters of chiral compounds (RRR- vs. all-rac-α tocopherol) by global gene expression profiling

Summary Pharmacologically active compounds (e.g. from the groups of pharmaceutical drugs, cofactors or vitamins) often consist of two or more stereoisomers (enantiomers or diastereoisomers) which may differ in their pharmacodynamic/kinetic, toxicological and biological properties. A well-known example is vitamin E which is predominantly administered as two different forms, one derived from natural sources (mainly soybeans), and one from production by chemical total-synthesis. While vitamin E from natural sources occurs as a single stereoisomer ( RRR - α -tocopherol), synthetic vitamin E ( all-rac - α -tocopherol) is an equimolar mixture of eight stereoisomers. Based on a number of animal studies it has been suggested that the biological potency of natural-source vitamin E is 1.36 greater compared to its counterpart produced by chemical synthesis. In this study, we have used the Affymetrix GeneChip ® technology to evaluate the feasibility of a new bio-assay where the gene regulatory activities of RRR - α -tocopherol and all-rac - α -tocopherol were quantified and compared on the genome-wide level. For this purpose, HepG2 cells were supplemented with increasing amounts of RRR - or all-rac - α -tocopherol for 7 days. Genes showing a dose-related induction/repression were identified by global gene expression profiling. Our findings show that RRR - and all-rac - α -tocopherol share an identical transcriptional activity, i.e. induce/repress the expression of the same set of genes. Based on the transcriptional dose–response data, EC50 and IC50 values were determined for each of these genes. The feasibility of calculating a “transcriptional potency factor” of RRR - vs. all-rac - α -tocopherol was evaluated by dividing the EC50/IC50 of RRR - α -tocopherol by the corresponding EC50/IC50 of all-rac - α -tocopherol for every of the vitamin E responsive genes. Using this approach we have calculated 215 single biopotency ratios. Subsequently, the mean of all potency ratios was found to be 1.05. In the present work we propose a new assay for the analysis and comparison of the biological activity and potency of chiral compounds in vivo.