Determination of drug stability against human liver microsomes by utilization of short-end injection capillary electrophoresis mode
| Authors | |
|---|---|
| Year of publication | 2010 |
| Type | Article in Proceedings |
| Conference | Final Program & Book of Abstracts, 25th International Symposium on Microscale BioSeparations MSB 2010 |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | capillary electrophoresis, drug stability, HLM |
| Description | During a new drug development, the pharmaceutical companies endeavour to predict action of enzymes on the given new drug candidate within human body. This knowledge enables to omit the substances with inappropriate qualities and thus simplify implementation of the following tests. The current methods of drug stability assessment based on incubations with whole hepatocytes or even liver slices are laborious and time-consuming nevertheless. In this regard, the human liver microsomes (HLM) provide possibility of unfavourable state circumvent, because they offer advantage of fast analysis combined with preservation of realistic simulation of conditions inside the liver. The goal of this study was to introduce a new method based on the capillary electrophoresis (CE) allowing assessment of new drug candidate stability against HLM. Since, HLM contain high concentrations of membrane-bond proteins, the short-end injection CE mode was used to reduce the time spend by sample inside the capillary. A replaceable polymeric gel as a background electrolyte was used either to avoid the adsorption of sample components onto inner capillary wall in order to ensure high reproducibility of the method. Considering the large variety of possible new drug candidates, the method based on NADP production monitoring was hired rather than usually used measurement of substrate consumption or product creation, respectively. In this manner, the method allows analysis of every compound with does not co-migrate with NADP. As result the generic method enabling fast determination of new drug candidate stability was established. The method's validation showed an RSD of 1.77 % for migration time (n = 6), an RSD of 2.04 % for relative peak areas (n = 6) for NADP. Finally, method's potentiality was proved by carrying out of fast screen of 12 chosen probe drugs. |
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