On-line version ISSN 0717-7712
GUTIERREZ-CORREA, JOSÉ and STOPPANI, ANDRÉS O.M.. Trypanosoma cruzi DIHYDROLIPOAMIDE DEHYDROGENASE IS INACTIVATED BY PHENOTHIAZINES IN THE PRESENCE OF CYTOCHROME C AND HYDROGEN PEROXIDE: EFFECTS OF ANTIOXIDANTS . Parasitol. latinoam. [online]. 2005, vol.60, n.3-4, pp. 105-121. ISSN 0717-7712. http://dx.doi.org/10.4067/S0717-77122005000200001.
Cytochrome c catalyzed the oxidation of phenothiazines (PTZ) in the presence of hydrogen peroxide. The transient formation of the promazine radical cation (PZ+.) has been demonstrated by light absorption measurements as well as by its conversión to promazine sulfoxide. Trypanosoma cruzi dihydrolipoamide dehydrogenase (LADH T c) was irreversibly inhibited by treatment with cytochrome c (cyt c)/H2O2 system supplemented with PTZ. LADH T c inactivation depended on a) The PTZ structure b) Time of incubación with the complete oxidant system c) The presence of an antioxidant that intercept free radicals. PZ, thioridazine (TRDZ) and trimeprazine (TMPZ), were the most effective systems out of twelve PTZ studied, with inactivation values of 82, 76 and 72%, respectively, after 90 min of incubation. LADH T c inactivation by PZ (with alkylamine substituent at N 10 position) decreased by its structural modification at 2 position (inactivation PZ > chlorpromazine (CPZ) > propionylpromazine (PPZ)>trifluopromazine (TFPZ)) or at N 10 position (inactivation PZ > TMPZ > promethazine (PMTZ)) PTZ activity with piperidinyl substituent at N10 position depended on the group at 2 position (TRDZ, with thiomethyl group, has high inactivating effect on LADH T c; propericyazine (PCYZ), with cyano group, is much less active). Apparently, piperazinyl substituent at the N10 position on the phenothiazine have not an important function in the compound's inactivating effect on LADH T c. The effect of PTZ with Cl at 2 position (CPZ, prochlorperazine (PCP), perphenazine (PFZ)) was higher than the effect of compounds with CF3 in the same position (TFPZ,trifluoperazine (TFP),fluphenazine (FFZ) ) independent on the structure of substituents at N10 position. Production of PTZ+. radicals was essential for LADH T c inactivation and this effect depended on the stability of these free radicals. Comparision of inactivation values for LADH T c and mammalian LADH demonstrated a greater sensitivity of LADH T c to various PTZ studied. Thiol compounds (such as GSH and N-acetylcysteine), tyrosine, tryptophan, NADH, ascorbate and trolox prevented LADH T c inactivation by the cyt c/H2O2/PTZ systems in agreement with their ability for to suppress PTZ+. radicals. The role of PTZ+. as enzyme inhibitors, or as generators of secondary free radicals and metabolite depletors for phenothiazines cytotoxicity is discussed
Keywords : Phenothiazines; Trypanosoma cruzi; Dihydrolipoamide Dehydrogenase; Cytochrome c.