Replacement of a Thiourea with an Amidine Group in a Monofunctional Platinum–acridine Antitumor Agent. Effect on DNA Interactions DNA Adduct Recognition and Repair

摘要

A combination of biophysical, biochemical, and computational techniques was used to delineate mechanistic differences between the platinum–acridine hybrid agent [PtCl(en)(L)](NO3)2 (complex >1, en = ethane-1,2-diamine, L = 1-[2-(acridin-9-ylamino)ethyl]-1,3- dimethylthiourea) and a considerably more potent second-generation analogue containing L′ = N-[2-(acridin-9-ylamino)ethyl]-Nmethylpropionamidine (complex >2). Calculations at the density functional theory level provide a rationale for the binding preference of both complexes for guanine-N7 and the relatively high level of adenine adducts observed for compound >1. A significant rate enhancement is observed for binding of the amidine-based complex >2 with DNA compared with the thiourea-based prototype >1. Studies conducted with chemical probes and on the bending and unwinding of model duplex DNA suggest that adducts of complex >2 perturb B-form DNA more severely than complex >1, however, without denaturing the double strand and significantly less than cisplatin. Circular and linear dichroism spectroscopies and viscosity measurements suggest that subtle differences exist between the intercalation modes and adduct geometries of the two complexes. The adducts formed by complex >2 most efficiently inhibit transcription of the damaged DNA by RNA polymerase II. Not only do complexes >1 and >2 cause less distortion to DNA than cisplatin, they also do not compromise the thermodynamic stability of the modified duplex. This leads to a decreased or negligible affinity of HMG domain proteins for the adducts formed by either Pt-acridine complex. In a DNA repair synthesis assay the lesions formed by complex >2 were repaired less efficiently than those formed by complex >1. These significant differences in DNA adduct formation, structure, and recognition between the two acridine complexes and cisplatin help to elucidate why compound >2 is highly active in cisplatin-resistant, repair proficient cancer cell lines.