Screening Of Drug Stability Against Microsomes By Means Of Capillary Electrophoresis
| Title in English | Application of sweeping - MEKC combination for monitoring of diclofenac reaction with cytochrome P450 2C9 isoform |
|---|---|
| Authors | |
| Year of publication | 2009 |
| Type | Article in Proceedings |
| Conference | Book of Abstracts 15th American symposium on Biotechnology, Biomedical and Pharmaceutical and Industrial Application of CE and Microchip Technology |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | drug stability; microsomes; capillary electrophoresis |
| Description | Today, most pharmaceutical companies employ a general paradigm in drug development, which incorporates a sub-loop termed "developability screening and predictions" into the classical cycle of biology and chemistry. The goal is to do a better job selecting candidate drugs early in the development process to avoid problems later on in clinical trials, when more resources have been invested. Among others stability against cytochrome P450 enzymes (CYP) constitutes one of the most important parameters of the new drug candidates. To gain a proper view of drug stability in the human body, the use of human liver microsomes (HLM) as a model system is preferred to a set of analyses with individual recombinant CYP isoforms. The aim of this study was to develop a generic method allowing assessment of candidate drug stability against HLM. Previously described NADP production monitoring [1] was used as a principle of the proposed method. All the experiments were carried out on the P/ACE TM MDQ CE system. An uncoated fused silica capillary with a total length of 50 cm (effective length 10 cm) and internal diameter 75 um was used as separation column. 10% (v/v) linear polyacrylamide in 50 mM disodium hydrogen phosphate was used as a background electrolyte. Separations were performed by application of 22 kV (positive polarity) while the capillary was thermostated at 37 C. Samples were injected by application of a pressure of 0.5 psi to the outlet vial for 3 s. Incubation mixtures containing appropriate concentration of probe drug, 1 mg/ml HLM, 0.3 mM NADPH and 0.2 mg/ml phthalic acid prepared in 10 mM phosphate buffer pH 7.4 were incubated at 37 C and 450 rpm for 30 minutes. Resulting method showed an RSD of 1.77 % for migration time and an RSD of 2.04 % for relative peak area of NADP+ (n = 6). The LOD of NADP+ was 6.5 uM (S/N = 3) and the LOQ was 20 uM (S/N = 10). Recovery of 100.63 -103.91 % was determined (n = 6). Finally, validated method was used to carry out stability screening of 12 chosen probe drugs. It proved methods capabilities as the universal tool in the new drug development. |
| Related projects: |