mesna and perfosfamide

To determine the effect of 4-hydroperoxycyclophosphamide (4OOH-CP) on the respiration of human sperm, and investigate the protective properties of mesna and WR-1065.. SUNY Upstate Medical University, Syracuse, NY.. Men (n = 12) visited the Andrology Department for fertility evaluation.. None.. Sperm respiration.. Immediate decline in the rate of respiration was observed when 4OOH-CP was added to washed sperm or semen. The inhibition was concentration dependent. The respiration was less affected when 4OOH-CP was added to semen, suggesting the presence of protective factors in the seminal plasma. Excess of mesna or WR-1065 ameliorated the effect of 4OOH-CP. Mesna was the more potent of the two compounds. 4OOH-CP also inhibited the respiration of mitochondria from beef heart.. These findings emphasize the adverse effects of alkylating agents on sperm function. The results also provide a framework for thiol drug administration with high-dose alkylating agents to protect male fertility. The protective capacity of seminal plasma deserves further testing.

Topics: Alkylating Agents; Cyclophosphamide; Humans; Male; Mercaptoethylamines; Mesna; Mitochondria; Oxygen Consumption; Protective Agents; Semen; Spermatozoa

The kinetics of the reactions of glutathione (GSH) with 4-hydroperoxycyclophosphamide (4OOH-CP) and acrolein, a metabolite of 4OOH-CP, were investigated in a cell-free medium (pH approximately 7.5) and peripheral blood mononuclear cells. The ability of the thiol drugs, sodium 2-mercaptoethane sulfonate (mesna) and S-2-(3-aminopropylamino)ethanethiol (WR-1065), to affect the reactions of cellular GSH with the alkyalting agents was also studied. The amount of unreacted thiols in the various reactions was determined by derivatization with monobromobimane, followed by separation of fluorescent-labeled thioether adducts using high-pressure liquid chromatography. The second-order rate constants (k(2)) for reactions of GSH, mesna, and WR-1065 with 4OOH-CP in solution were 38 +/- 5, 25 +/- 5, and 880 +/- 50 M(-1) s(-1), respectively. The corresponding k(2) for reactions of GSH, mesna, and WR-1065 with acrolein were 490 +/- 100, 700 +/- 150, and >2000 M(-1) s(-1), respectively. The apparent rate constants for reactions of cellular GSH with acrolein and 4OOH-CP were smaller than those obtained in solution. Assuming that the k(2) is the same inside and outside cells, we estimate the first-order rate constant (k(1)) for transfer of 4OOH-CP and acrolein across the cell membrane as approximately 0.01 and approximately 0.04 s(-1), respectively. WR-1065 was more effective than mesna in blocking depletion of cellular GSH (because it passes into the cell more quickly and has higher reaction rates with the alkylators than the latter compound). When WR-1065 and mesna were used together, the protection against cellular depletion of GSH was additive. Our results are relevant to the administration of thiol drugs with high-dose alkylating agents.

Topics: Acrolein; Antineoplastic Agents, Alkylating; Cell Membrane; Cyclophosphamide; Glutathione; Humans; In Vitro Techniques; Kinetics; Leukocytes, Mononuclear; Mercaptoethylamines; Mesna; Protective Agents

Cyclophosphamide (CY) is metabolized to 4-hydroxy-CY which spontaneously breaks down to the reactive intermediates, phosphoramide mustard (PAM) and acrolein. The alkylating property of PAM is thought to mediate the anti-proliferative and cytotoxic actions of CY. Acrolein is known to bind sulfhydryl groups of cellular molecules and may contribute to the action of CY. However, the role of acrolein in the CY-induced immunosuppression remains unclear. The results of studies in which a hepatocyte co-culture system was used suggest that acrolein may play an important role in the cytotoxic action of CY, whereas those investigations using activated derivatives of CY indicate that acrolein is not an important factor. Thus, both approaches of CY exposure were utilized in the present study. Splenocytes of B6C3F1 mice were incubated with syngeneic isolated hepatocytes and CY or with 4-hydroperoxycyclophosphamide (4-HC) (which spontaneously decomposes to 4-hydroxy-CY). The in vitro antibody forming cell (AFC) response was found to be suppressed with both methods of exposure to CY metabolites. The addition of DNA to bind extracellular PAM was ineffective in preventing the suppression produced by hepatocyte-activated CY. However, it was also observed that DNA was able to attenuate the PAM-induced suppression. The sulfhydryl compounds 2-mercaptoethanesulfonate (MESNA) (15 microM) or reduced glutathione (GSH) (1 mM) inhibited the suppression of the AFC response of splenocytes incubated with CY and mouse hepatocytes. The suppression produced by 4-HC, however, was not affected by MESNA and only slightly inhibited by GSH. Similarly, the PAM-induced suppression was not affected by MESNA and slightly attenuated with GSH. In contrast, both MESNA and GSH were very effective in abrogating the acrolein-induced suppression, whereas DNA was ineffective. The findings of this study suggest that in the hepatocyte co-culture system, the immunosuppressive actions of CY are mediated by acrolein generated outside of the splenocyte, whereas the 4-HC induced suppression is not mediated by extracellular acrolein. Thus, this difference may explain the contradictory findings of previous studies that used different means of exposing cells to activated CY.

Topics: Acrolein; Animals; Antibody Formation; Cell Survival; Cells, Cultured; Cyclophosphamide; DNA; Female; Liver; Mesna; Mice; Spleen

Topics: Animals; Cricetinae; Cricetulus; Cyclophosphamide; Cysteine; Drug Therapy, Combination; Female; Humans; Male; Mercaptoethanol; Mesna; Mice; Mice, Inbred C57BL