Characterization of cisplatin-glutathione adducts by liquid chromatography-mass spectrometry evidence for their formation in vitro but not in vivo after concomitant administration of cisplatin and glutathione to rats and cancer patients

Bernareggi, A.; Torti, L.; Facino, Roberto Maffei; Carini, Marina; Depta, Gabriele; Casetta, Bruno; Farrell, Nicholas; Spadacini, Sabina; Ceserani, R.; Tognella, S

doi:10.1016/0378-4347(95)00098-4

Cited by 41 publications

(33 citation statements)

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“…Bernareggi and coworkers identified a monoplatinum-GSH conjugate that formed spontaneously in solution under incubation conditions that differed slightly from those used in this study (Bernareggi et al, 1995). The MS fragmentation profile and structure proposed by Bernareggi are the same as those of GSH-2 identified in this study.…”

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confidence: 64%

“…Formation of platinum-GSH conjugates and platinum-cysteine conjugates has been demonstrated in several laboratories (Ishikawa and Ali-Osman, 1993;Bernareggi et al, 1995). Bernareggi and coworkers identified a monoplatinum-GSH conjugate that formed spontaneously in solution under incubation conditions that differed slightly from those used in this study (Bernareggi et al, 1995).…”

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confidence: 58%

“…We propose that the initial step in the activation pathway is the formation of a GSH-platinum conjugate. GSH conjugates of cisplatin have been shown to form spontaneously in solution and have been isolated from the kidneys of rats treated with cisplatin (Mistry et al, 1989;Ishikawa and Ali-Osman, 1993;Bernareggi et al, 1995). However, these GSH-platinum conjugates have been proposed to be inactive forms of the drug and have not been implicated in the nephrotoxicity.…”

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High Pressure Liquid Chromatography and Mass Spectrometry Characterization of the Nephrotoxic Biotransformation Products of Cisplatin

et al. 2003

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This article is available online at http://dmd.aspetjournals.org ABSTRACT:Previous studies have shown that cisplatin requires metabolic activation to become nephrotoxic. The activation is proposed to be via the metabolism of a glutathione-platinum conjugate to a cysteinylglycine-platinum conjugate, which is further processed to a cysteine conjugate. Preincubating cisplatin with glutathione (GSH), cysteinylglycine, or N-acetylcysteine (NAC) results in a transient increase in the toxicity of cisplatin toward renal proximal tubular cells. In this study, the preincubation solutions were analyzed by high pressure liquid chromatography (HPLC), atomic absorption spectrometry, and mass spectrometry (MS) to characterize the formation and structure of the platinum conjugates. HPLC analysis of the cisplatin-GSH, cisplatin-cysteinyl-glycine, and cisplatin-NAC preincubation solutions revealed two new platinum-containing peaks in each of the solutions. MS-MS analysis of the peaks revealed a diplatinum-and a monoplatinum conjugate in each of the solutions. Analysis of the composition and toxicity of the solutions with time showed that the transient increase in toxicity correlated with the formation of the monoplatinum conjugate whereas prolonged preincubation decreased toxicity and correlated with the formation of the diplatinum conjugate. The monoplatinum-monoglutathione conjugate is a substrate for ␥-glutamyl transpeptidase, an enzyme that is essential for the nephrotoxicity of cisplatin. The monoplatinum-mono-NAC conjugate can be deacetylated to a cysteine conjugate, which is a substrate for pyroxidol phosphate (PLP)-dependent cysteine S-conjugate ␤-lyase. This PLPdependent enzyme is proposed to catalyze the final step in the metabolic activation of cisplatin. Identification of the structure and toxicity of these conjugates further elucidates the metabolism of cisplatin to a nephrotoxin.The use of cisplatin in the treatment of ovarian, germ-cell, head and neck, bladder, and other tumors is limited by its nephrotoxicity. Cisplatin kills dividing cells by forming platinum-DNA cross-links that prevent DNA synthesis and result in cell death (O'Dwyer et al., 1999). In the kidney, cisplatin is toxic to the proximal tubular cells, which are not replicating, suggesting an alternative mechanism of toxicity. Daley-Yates and McBrien showed the first evidence that indicated biotransformation products of cisplatin were the nephrotoxic compounds (Daley-Yates and McBrien, 1984). These investigators reported that seven platinum containing species were present in plasma that could be separated via HPLC 2 following a single dose of cisplatin. The mixture of platinum-containing species was injected into rats and was more nephrotoxic than cisplatin. However, the anti-cancer activity of the mixture of platinum-containing species was less effective than cisplatin in a mouse leukemia model. The mechanism of cisplatin-induced nephrotoxicity was not identified until studies in our laboratory demonstrated that cisplatin-induced renal toxicity is due...

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confidence: 64%

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High Pressure Liquid Chromatography and Mass Spectrometry Characterization of the Nephrotoxic Biotransformation Products of Cisplatin

et al. 2003

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“…One of the direct actions of CDDP is the formation of CDDP-glutathione conjugate, 32,33 which is subsequently metabolized to CDDP-cysteinyl-glycine conjugates via ␥-glutamyl transpeptidase in the proximal tubules. The conjugates may be further cleaved to cysteine-conjugates by aminopeptidase, which is also on the surface of the cells.…”

Section: Discussionmentioning

confidence: 99%

Lack of the Growth Factor Midkine Enhances Survival against Cisplatin-Induced Renal Damage

Kawai

Sato

Yuzawa

et al. 2004

The American Journal of Pathology

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Although cisplatin acts directly on proximal tubule epithelial cells and causes cell death, little is known regarding the biological significance of its secondary effects, such as inflammation. The growth factor midkine is highly expressed in the proximal tubule and exerts ambivalent activities as to cisplatin nephrotoxicity, ie, anti-apoptotic and chemotactic ones. Here we report that midkine-deficient mice show a significantly higher survival rate than wild-type mice. The levels of blood urea nitrogen and tubular degeneration and apoptosis were higher in wild-type mice despite the anti-apoptotic activity of midkine. We found that recruitment of neutrophils was more enhanced in wild-type mice, this being consistent with the chemotactic activity of midkine. Midkine expression in wild-type mice persisted for 24 hours, and then dramatically decreased. Preadministration of midkine anti-sense oligodeoxyribonucleotide to wildtype mice suppressed midkine expression, and consequently neutrophil infiltration. It is of note that neutrophil infiltration, apoptosis, and elevation of blood urea nitrogen became conspicuous sequentially, namely 1, 2, and 3 days after cisplatin administration, respectively. These findings suggest that early molecular events involving midkine induce inflammatory response and their circuits eventually enhance the death of the proximal tubule epithelial cells. The results indicate the crucial role of inflammation in cisplatin-induced renal damage, and provide a candidate molecular target for its prevention. Cisplatin [cis-dichlorodiammine-platinum (II), CDDP] is widely used to treat carcinoma patients. However, its full clinical utility is significantly limited because of its nephrotoxicity. The main site of CDDP action is in proximal tubule epithelial cells. In proliferating cancer cells, CDDP inhibits DNA and RNA synthesis. In contrast, the effects of CDDP on nonproliferating cells, such as proximal tubule epithelial cells, can be categorized into mainly two primary types.

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“…The model included passage of CDDP through the cell membrane, binding of CDDP to cellular thiols, and platination of DNA. We also studied the reaction of GSH with 4-hydroperoxycyclophosphamide and acrolein ; the latter is a toxic metabolite of cyclophosphamide.Although CDDP reaction with thiols has been studied extensively (Berners-Price and Kuchel, 1990a,b;Ishikawa and Ali-Osman, 1993;Bernareggi et al, 1995;Perez-Benito et al, 1995;Bose et al, 1997;El-Khateeb et al, 1999), there is little kinetic data under clinically relevant conditions. In this work, we use UV absorption, HPLC, and atomic absorption spectroscopy (AAS) to measure the rates of CDDP reaction with GSH, WR-1065, and mesna, using a therapeutic con-1 Abbreviations used are:…”

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Kinetic Study of the Reaction of Cisplatin with Thiols

Dabrowiak¹,

Goodisman²,

Souid³

2002

Drug Metab Dispos

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ABSTRACT:The reactions of cisplatin [cis-diamminedichloroplatinum(II), CDDP] with glutathione (GSH) and drug thiols were investigated at 37°C in 100 mM Tris-NO 3 , pH ϳ7.4, using a clinically relevant concentration of CDDP (33 ⌴), 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. CDDP 1 exerts its antitumor activity by binding to cellular DNA (Rosenberg, 1971;Gelasco and Lippard, 1999). When the drug enters the cell, it passes through the cytosol, enters the nuclear envelope, binds to nitrogen atoms on the bases of DNA, and promotes cell death by apoptosis (Bellon et al., 1991;Demarcq et al., 1994). The cytotoxic activity of CDDP thus seems to correlate with the amount of platinum (Pt) bound to DNA (Zwelling et al., 1979;Lindauer and Holler, 1996).Biological thiols such as GSH, metallothionine, and other protein thiols defend the cell against CDDP (Kraker et al., 1985;Pattanaik et al., 1992;Ishikawa and Ali-Osman, 1993;Reedijk and Teuben, 1999). Pt ions entering the cell may preferentially bind to GSH and metallothionine (Dedon and Borch, 1987), both present in millimolar concentrations in the cytoplasm (Souid et al., 1999(Souid et al., , 2001; A.-K. Souid personal observation). Because Pt-thiol adducts interact less well with DNA (Volckova et al., 2002), formation of these complexes limits the amount of drug available for binding to DNA. Furthermore, continued exposure to CDDP can increase the cytosolic sulfhydryl level (Schilder et al., 1990;Eastman 1991;Godwin et al., 1992), which may produce CDDP resistance.The sulfhydryl agents WR-1065 and mesna are often administered to mitigate toxicities of Pt-based compounds and alkylating agents, such as cyclophosphamide (Brock et al., 1982;Souid et al., 2001;Tacka et al., 2002). The protective mechanism of WR-1065 and mesna involves reaction of the thiolate ion with CDDP in a manner analogous to that of GSH (Leeuwenkamp et al., 1991;Treskes et al., 1991).Previously, we measured the rate of CDDP binding to DNA in peripheral blood mononuclear cells and ovarian cancer cells (Sadowitz et al., 2002). By blocking the cellular thiol groups and by adding known concentrations of WR-1065 or mesna, we showed how the amount of CDDP reaching the DNA is affected by thiol concentration. For example, blocking all cellular thiol groups with N-ethylmaleimide increased DNA platination by ϳ8-fold. These data were used to generate a simple kinetic model that reproduced the measured Pt bound to DNA as a function of incubation time and CDDP and thiol concentrations. The model included passage of CDDP through the cell membrane, binding of CDDP to cellular thiols, and platination of DNA. We also studied the reaction of GSH with 4-hydroperoxycyclophosphamide and acrolein ; the latter is a toxic metabolite of cyclophosphamide.Although CDDP reaction with thiols has been studied ex...

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Characterization of cisplatin-glutathione adducts by liquid chromatography-mass spectrometry evidence for their formation in vitro but not in vivo after concomitant administration of cisplatin and glutathione to rats and cancer patients

Cited by 41 publications

References 20 publications

High Pressure Liquid Chromatography and Mass Spectrometry Characterization of the Nephrotoxic Biotransformation Products of Cisplatin

High Pressure Liquid Chromatography and Mass Spectrometry Characterization of the Nephrotoxic Biotransformation Products of Cisplatin

Lack of the Growth Factor Midkine Enhances Survival against Cisplatin-Induced Renal Damage

Kinetic Study of the Reaction of Cisplatin with Thiols

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