Reaction of polynuclear platinum antitumor compounds with reduced glutathione studied by multinuclear (1H, 1H-15N gradient heteronuclear single-quantum coherence,and 195Pt) NMR spectroscopy

A possible explanation for the low bioavailability of platinum antitumor compounds is their high reactivity with the sulfur-containing tripeptide glutathione (GSH; deprotonated GSH = SG). GSH is located in the intracellular matrix of the cell with a normal concentration of 5-10 mM. In vivo, only a small fraction of the administered drug will migrate into the cell, resulting in relatively high concentrations of GSH compared to that of the drug. The products of the reactions of trans-PtCl(NH(3))(2)](2)-mu-trans-Pt(NH(3))(2)(NH(2)(CH(2))(6)NH(2))(2)]](NO(3))(4) (BBR3464; 1,0,1/t,t,t, n = 6), trans-PtCl(NH(3))(2)](2)-mu-(H(2)N(CH(2))(6)NH(2))](NO(3))(2) (BBR3005; 1,1/t,t, n = 6), trans-PtCl(NH(3))(2)](2)-mu-(H(2)N(CH(2))(3)NH(2)(CH(2))(4)NH(2))]Cl(3) (BBR3571; 1,1/t,t-spermidine, n = 3, 4), and trans-PtCl(2)(NH(3))(2)] (t-DDP) with reduced GSH in phosphate-buffered saline (pH 7.35) have been characterized by (1)H, (195)Pt, and (1)H(-)(15)N gradient heteronuclear single-quantum coherence NMR spectroscopy and high-performance liquid chromatography (HPLC) coupled with electrospray ionization time-of-flight mass spectrometry to determine likely metabolites of the complexes with GSH. Chemical shifts (NMR) and retention times (HPLC) established via analysis of the t-DDP profile served as a fingerprint to compare results obtained for the products afforded by the degradation of the polynuclear compounds by GSH. Identical kinetic profiles and chemical shifts between the metabolites and the t-DDP/GSH products allowed identification of the final product for the 1:2 Pt:GSH reaction as a dinuclear species trans-Pt(SG)(NH(3))(2)](2)-mu-SG], in which glutathione bridges the two platinum centers via only the sulfur atom.