In vitro characterization of a novel C,N-cyclometalated benzimidazole Ru(II) complex: stability, intracellular distribution and binding, effects on organic osmolyte homeostasis and induction of apoptosis

TY - JOUR

T1 - In vitro characterization of a novel C,N-cyclometalated benzimidazole Ru(II) complex

T2 - stability, intracellular distribution and binding, effects on organic osmolyte homeostasis and induction of apoptosis

AU - Dam, Celina Støving

AU - Henarejos, Sergio Alejo Perez

AU - Tsolakou, Theodosia

AU - Segato, Christian Alexander

AU - Gammelgaard, Bente

AU - Yellol, Gorakh S.

AU - Ruiz, José

AU - Lambert, Ian Henry

AU - Stürup, Stefan

PY - 2015/5/1

Y1 - 2015/5/1

N2 - In the present work a novel C,N-cyclometalated benzimidazole Ru(ii) arene complex (GY34) was characterized by applying an alternative, diverse approach considering both chemical and biological aspects. RP-HPLC-ICP-MS and RP-HPLC-ESI-MS analysis proved that GY34 in both RPMI-1640 cell medium and ammonium acetate buffer was transformed into several subspecies and the importance of evaluating and controlling analyte stability throughout experiments was demonstrated. Applying a novel cell fractionation protocol GY34 was found to target cell nuclei and mitochondria in Ehrlich Lettré Ascites (ELA) cells, with the intracellular distribution depending on GY34 concentration in the cell medium during incubation. In ELA cells 96 ± 0.2% of cytosolic GY34 was bound to high-molecular species. Furthermore, using the tracer technique GY34 was found to reduce uptake and increase release of the organic osmolyte taurine in ELA cells, with innate resistance to Cisplatin and in A2780 human ovarian cancer cells, with acquired resistance to Cisplatin. Importantly, FACS analysis revealed that GY34 induced apoptosis in ELA cells. The present data suggest the potential of GY34 in overcoming Cisplatin resistance. The methodology applied can be used as a general protocol and an additional tool in the initial evaluation of novel metal-based drugs.

AB - In the present work a novel C,N-cyclometalated benzimidazole Ru(ii) arene complex (GY34) was characterized by applying an alternative, diverse approach considering both chemical and biological aspects. RP-HPLC-ICP-MS and RP-HPLC-ESI-MS analysis proved that GY34 in both RPMI-1640 cell medium and ammonium acetate buffer was transformed into several subspecies and the importance of evaluating and controlling analyte stability throughout experiments was demonstrated. Applying a novel cell fractionation protocol GY34 was found to target cell nuclei and mitochondria in Ehrlich Lettré Ascites (ELA) cells, with the intracellular distribution depending on GY34 concentration in the cell medium during incubation. In ELA cells 96 ± 0.2% of cytosolic GY34 was bound to high-molecular species. Furthermore, using the tracer technique GY34 was found to reduce uptake and increase release of the organic osmolyte taurine in ELA cells, with innate resistance to Cisplatin and in A2780 human ovarian cancer cells, with acquired resistance to Cisplatin. Importantly, FACS analysis revealed that GY34 induced apoptosis in ELA cells. The present data suggest the potential of GY34 in overcoming Cisplatin resistance. The methodology applied can be used as a general protocol and an additional tool in the initial evaluation of novel metal-based drugs.

U2 - 10.1039/c5mt00056d

DO - 10.1039/c5mt00056d

M3 - Journal article

C2 - 25805368

SN - 1756-5901

SP - 885

EP - 895

JO - Metallomics

JF - Metallomics

IS - 5

ER -