Kinetic study of the reaction of cisplatin with thiols.

Abstract

The reactions of cisplatin [cis-diamminedichloroplatinum(II), CDDP] with glutathione (GSH) and drug thiols were investigated at 37 degrees C in 100 mM Tris-NO(3), pH approximately 7.4, using a clinically relevant concentration of CDDP (33 micro M), a large excess of GSH (16.5 mM), and [NaCl] of 4.62 mM. The conditions were designed to mimic passage of CDDP through the cytosol. The reactions were studied by UV-absorption spectroscopy, high-pressure liquid chromatography (HPLC), and atomic absorption spectroscopy. The initial rates, detected by UV absorbance, confirmed that the reactions are first order in [CDDP]. The HPLC peak corresponding to CDDP was analyzed for platinum content by atomic absorption spectroscopy, which decreased exponentially with time, confirming that the reactions are first order in [CDDP] and allowing determination of the pseudo first order rate constants (k(1)). For reaction of the dichloro form of CDDP with GSH, the k(1) value was approximately 2.2 x 10(-4) s(-1) (t(1/2) of approximately 53 min), giving the second order rate constant value (k(2)) of approximately 1.3 x 10(-2) M(-)1 s(-1). Reaction of a mixture of the aquated forms of CDDP with GSH gave a lower k(1) value ( approximately 0.9 x 10(-4) s(-1)). Reaction of CDDP with sodium 2-mercaptoethanesulfonate (mesna) gave a k(1) value of approximately 1.8 x 10(-4) s(-1) (t(1/2) of approximately 65 min and k(2) of approximately 1.1 x 10(-2) M(-1) s(-1)). Reaction of CDDP with S-2-(3-aminopropylamino)ethanethiol (WR-1065) gave a k(1) value of approximately 12.0 x 10(-4) s(-1) (t(1/2) of approximately 10 min and k(2) of approximately 7.3 x 10(-2) M(-)1 s(-1)). The relatively slow reaction rate of CDDP with GSH is consistent with the efficient DNA platination by CDDP in the presence of millimolar concentration of GSH in the cytosol.