Hepatotoxicity in Mice of a Novel Anti-parasite Drug Candidate Hydroxymethylnitrofurazone: A Comparison with Benznidazole

Abstract

BackgroundTreatment of Chagas disease, caused by Trypanosoma cruzi, relies on nifurtimox and benznidazole (BZL), which present side effects in adult patients, and natural resistance in some parasite strains. Hydroxymethylnitrofurazone (NFOH) is a new drug candidate with demonstrated trypanocidal activity; however, its safety is not known.MethodsHepG2 cells dose response to NFOH and BZL (5–100 µM) was assessed by measurement of ROS, DNA damage and survival. Swiss mice were treated with NFOH or BZL for short-term (ST, 21 d) or long-term (LT, 60 d) periods. Sera levels of cellular injury markers, liver inflammatory and oxidative stress, and fibrotic remodeling were monitored.ResultsHepG2 cells exhibited mild stress, evidenced by increased ROS and DNA damage, in response to NFOH, while BZL at 100 µM concentration induced >33% cell death in 24 h. In mice, NFOH ST treatment resulted in mild-to-no increase in the liver injury biomarkers (GOT, GPT), and liver levels of inflammatory (myeloperoxidase, TNF-α), oxidative (lipid peroxides) and nitrosative (3-nitrotyrosine) stress. These stress responses in NFOH LT treated mice were normalized to control levels. BZL-treated mice exhibited a >5-fold increase in GOT, GPT and TNF-α (LT) and a 20–40% increase in liver levels of MPO activity (ST and LT) in comparison with NFOH-treated mice. The liver inflammatory infiltrate was noted in the order of BZL>vehicle≥NFOH and BZL>NFOH≥vehicle, respectively, after ST and LT treatments. Liver fibrotic remodeling, identified after ST treatment, was in the order of BZL>vehicle>NFOH; lipid deposits, indicative of mitochondrial dysfunction and in the order of NFOH>vehicle>BZL were evidenced after LT treatment.ConclusionsNFOH induces mild ST hepatotoxicity that is normalized during LT treatment in mice. Our results suggest that additional studies to determine the efficacy and toxicity of NFOH are warranted.

Clomipramine and Benznidazole Act Synergistically and Ameliorate the Outcome of Experimental Chagas Disease

“…*, P Ͻ 0.05; **, P Ͻ 0.01; ***, P Ͻ 0.001; ns, no significant difference. tions (16), while the presence of CMP seemed not to induce additional apparent injuries (23).…”

“…Considering the side effects of BZ, in particular its hepatic intolerance and hepatotoxicity due to the inhibition of DNA and protein synthesis in hepatocytes (16,42), we evaluated the toxicities of the two drugs at the selected doses. Even though the differences in the indicators measured (liver weight and plasma GPT levels) were not statistically significant, it is important to stress that both treatments showed a moderate increase in GPT levels compared with those in the control group receiving drinking water by gavage (INT).…”

“…Samples of blood were centrifuged at 2,500 rpm for 5 min, and the plasma was analyzed in order to determine the activities of glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) as liver injury biomarkers (16). On the other hand, creatine phosphokinase (total CK) and creatine phosphokinase myocardial band isoform MB (CK-MB) as a cardiac injury marker were tested (29).…”

“…The control group received water by oral gavage in place of drug treatment. We evaluated liver weight and plasma GPT levels, a specific liver injury biomarker (16).…”

“…The main side effect of BZ is hepatic intolerance or hepatotoxicity, usually concomitant with hypersensitivity reactions at the beginning of treatment, and medullar toxicity and peripheral neuropathies at the end of treatment (15)(16)(17)(18)(19).…”