The Schistosoma mansoni Cytochrome P450 (CYP3050A1) Is Essential for Worm Survival and Egg Development

Schistosomiasis affects millions of people in developing countries and is responsible for more than 200,000 deaths annually. Because of toxicity and limited spectrum of activity of alternatives, there is effectively only one drug, praziquantel, available for its treatment. Recent data suggest that drug resistance could soon be a problem. There is therefore the need to identify new drug targets and develop drugs for the treatment of schistosomiasis. Analysis of the Schistosoma mansoni genome sequence for proteins involved in detoxification processes found that it encodes a single cytochrome P450 (CYP450) gene. Here we report that the 1452 bp open reading frame has a characteristic heme-binding region in its catalytic domain with a conserved heme ligating cysteine, a hydrophobic leader sequence present as the membrane interacting region, and overall structural conservation. The highest sequence identity to human CYP450s is 22%. Double stranded RNA (dsRNA) silencing of S. mansoni (Sm)CYP450 in schistosomula results in worm death. Treating larval or adult worms with antifungal azole CYP450 inhibitors results in worm death at low micromolar concentrations. In addition, combinations of SmCYP450-specific dsRNA and miconazole show additive schistosomicidal effects supporting the hypothesis that SmCYP450 is the target of miconazole. Treatment of developing S. mansoni eggs with miconazole results in a dose dependent arrest in embryonic development. Our results indicate that SmCYP450 is essential for worm survival and egg development and validates it as a novel drug target. Preliminary structure-activity relationship suggests that the 1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethan-1-ol moiety of miconazole is necessary for activity and that miconazole activity and selectivity could be improved by rational drug design.

Author Summary Over 600 million people in endemic countries are at risk of contracting schistosomiasis, which results in over 200,000 deaths each year and significant illness to most people that are infected. There are concerns that the drug widely used for the treatment of schistosomiasis, praziquantel, may be losing efficacy due to evolution of drug resistant worms. Since the disease mainly affects the poor in developing countries, pharmaceutical companies have little interest in developing new drugs and none are currently being tested. In this paper we focus on a novel parasite protein, cytochrome P450, which we propose to be a new drug target. Worms are unusual in having only one cytochrome P450 gene; humans have 57 cytochrome P450 genes. By using reverse genetic and chemical approaches we found that the schistosome cytochrome P450 is essential for worm survival and egg development and, therefore, is an essential and druggable target. Drugs that target fungal cytochrome P450s and are already in use for treating several human diseases were identified as potential hits for further development for schistosomiasis treatment.