| Description |
The main purpose of this thesis project is to develop and validate a high-throughput protocol to test drug-drug interactions. As a model, tyrosine kinase inhibitors were tested with commonly used antibiotics in Philadelphia chromosome positive leukemia patients. Tyrosine kinase inhibitors (TKI's), such as imatinib mesylate and dasatinib, are currently the frontline therapeutic agents for patients with leukemia characterized with the presence of the Philadelphia chromosome (Ph+), generated through a translocation event between chromosomes 9 and 22 (t[9,22][q34;q11]). However, interactions between TKI's and medications commonly taken by patients with leukemia, including antibiotics, have not been significantly studied. In this thesis, a protocol was established using innovative, high-throughput technology to determine imatinib and dasatinib maximal inhibitory concentrations and possible interactions with drugs that are often administered concomitantly with TKI therapy, antibiotics in particular. K562 cells were cultured in the presence of TKI's and cell proliferation was measured. Inhibitory concentrations of the TKI's were determined. Imatinib at concentrations IC10, IC50, and IC90 were then co-incubated with co-medications at concentrations reflecting therapeutic exposure. Co-medications were gentamicin, ceftazidime, sulfamethoxazole, trimethoprim, and a combination of sulfamethoxazole and trimethoprim at a 1:23 ratio, respectively, to emulate the constituents of the antibiotic commonly known as Bactrim. The results determined that sulfamethoxazole and trimethoprim have no significant effect on K562 cell proliferation or the ability of imatinib to reduce K562 cell proliferation. Gentamicin and ceftazidime displayed no difference from control perturbation on imatinib efficacy. However, gentamicin and ceftazidime were found to have an effect on K562 proliferation without the addition of a TKI. Therefore, further investigation is necessary to understand the mechanism behind gentamicin and ceftazidime reducing K562 cell proliferation. This assay can also be used to assess for potential in vitro TKI-drug interactions using robust, high-throughput screening techniques. |
