The trans Influence in the Modulation of Platinum Anticancer Agent Biology: The Effect of Nitrite Leaving Group on Aquation, Reactions with S-Nucleophiles and DNA Binding of Dinuclear and Trinuclear Compounds

摘要

To examine the effect of leaving group and trans influence on the geenral reactivity of polynuclear platinum antitumor agents we investigated substitution of the chloride leaving groups with nitrite ion, which forms strong bonds to Pt. It was of interest to explore whether nitrite could be used to modulate biological properties of these agents, in particular the deactivating reactions that occur on reaction with S-nucleophiles, involving loss of linking diamine under the trans influence of sulfur. Reported herein is a study of the synthesis, aquation, DNA binding and reactions with glutathione (GSH), methionine (met) and acetylmethione (AcMet) of nitro derivatives of di- and trinuclear platinum antitumor compounds: To examine the effect of leaving group and trans influence on the general reactivity of polynuclear platinum antitumor agents we investigated substitution of the chloride leaving groups with nitrite ion, which forms strong bonds to Pt. It was of interest to explore whether nitrite could be used to modulate biological properties of these agents, in particular the deactivating reactions that occur on reaction with S-nucleophiles, nvolving loss of the linking diamine under the trans influence of sulfur. Reported herein is a study of the synthesis, aquation, DNA binding and reactions with glutathione (GSH), methionine (Met) and acetylmethione (AcMet) of nitrito derivatives of di- and trinuclear platinum antitumor compounds: [{trans-PtNO2-ACHTUNGTRENUNG(NH3)2}2ACHTUNGTRENUNG(m-NH2ACHTUNGTRENUNG(CH2)6NH2)]ACHTUNGTRENUNG(NO3)2 (1- NO2) and [{trans-PtNO2ACHTUNGTRENUNG(NH3)2}2(mtrans- PtACHTUNGTRENUNG(NH3)2ACHTUNGTRENUNG{NH2ACHTUNGTRENUNG(CH2)6NH2}2)]- ACHTUNGTRENUNG(NO3)4 (1'-NO2). {1H,15N}-HSQC NMR studies revealed that 1-NO2 is inert to aquation reactions, even after prolonged incubation at physiological pH. Monitoring of the interaction of 1-NO2 with the duplex 5'-d(ATATGTACATAT) 2 (I) showed only unreacted complex, consistent with activation by aquation being a requirement for covalent DNA binding. The reaction of 1-NO2 with GSH was studied by 1H, 195Pt, 15N and {1H,15N}-HSQC NMR spectroscopy. For the parent dichlorido compounds (1 and 1') substitution of chloride by GS leads to drug degradation involving liberation of the diamine linker. While the same final products trans-[Pt(SG)2ACHTUNGTRENUNG(NH3)2] (5) and trans-[{Pt(SG)ACHTUNGTRENUNG(NH3)2}2-m-SG] (6) are formed, different mechanisms are involved, consistent with the trans influence NO2 > Cl ; the half-life is slightly longer for 1-NO2 (1.8 h) compared with 1 (1.3 h). Identification of the intermediate trans-[PtACHTUNGTRENUNG(NH3)2ACHTUNGTRENUNG(NO2)(SG)] (4) shows that the nitrito group remains coordinated while the linker amine is substituted by coordination of GS , and then trans labilization of the nitrito group occurs leading to 5 and 6. Reaction of the trinuclear 1'-NO2 with GSH follows essentially the same reaction pathway. Reaction of 1-NO2 with Met and AcMet is much slower and only 20% liberated amine was observed after reaction with Met for 24 h at 37 8C. The final product from reaction with AcMet is trans-[PtACHTUNGTRENUNG(NH3)2ACHTUNGTRENUNG(NO2)- ACHTUNGTRENUNG(AcMet)], as in this case coordination of the S-nucleophile does not lead to trans labilization of the nitrito group.