Reaction of geldanamycin and C17-substituted analogues with glutathione: product identifications and pharmacological implications.
17-Dimethylaminoethylamino-17-demethoxygeldanamycin (DMAG) and 17-allylamino-17-demethoxygeldanamycin (17-AAG) are two derivatives of geldanamycin (GA) that are currently undergoing clinical evaluation as anticancer agents. These agents bind to heat shock protein 90 (hsp90), resulting in the destabilization of client proteins and inhibition of tumor growth. In a search for the mechanism of hepatotoxicity, which is a dose-limiting toxicity for these agents, we found that GA and its derivatives, 17-AAG and 17-DMAG, react chemically (i.e., nonenzymatically) with glutathione (GSH). A combination of liquid chromatography/electrospray ionization/mass spectrometry and nuclear magnetic resonance analyses were used to identify the product of this reaction as a GSH adduct in which the thiol group of GSH is substituted in the 19-position of the benzoquinone ring. The reaction proceeds rapidly with GA and 17-DMAG (half-lives of approximately 1.5 and 36 min, respectively) and less rapidly with 17-AAG and its major metabolite, 17-AG (half-lives of approximately 9.8 and 16.7 h). The reaction occurs at pH 7.0, 37 degrees C, and a physiological concentration of GSH, indicating that cellular GSH could play a role in modulating the cellular toxicity of these agents and therefore be a factor in their mechanism of differential toxicity. Moreover, reactions with thiol groups of critical cellular proteins could be important to the mechanism of toxicity with this class of anticancer agents.
Laboratory of Clinical Pharmacology, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland 20993-0002, USA.
Richard L
RL
Cysyk
Robert J
RJ
Parker
Joseph J
JJ
Barchi
Patricia S
PS
Steeg
Neil R
NR
Hartman
John M
JM
Strong
D000903:Antibiotics, Antineoplastic / Q000737:chemistry*; D016227:Benzoquinones; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D005978:Glutathione / Q000737:chemistry*; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry; D047029:Lactams, Macrocyclic; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D010710:Phosphates / Q000737:chemistry; D011809:Quinones / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry
D000903:Antibiotics, Antineoplastic; D016227:Benzoquinones; D002021:Buffers; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D010710:Phosphates; D011809:Quinones; D013438:Sulfhydryl Compounds; D005978:Glutathione; C001277:geldanamycin