A computational analysis of the binding mode of closantel as inhibitor of the Onchocerca volvulus chitinase: insights on macrofilaricidal drug design.

Onchocerciasis is a leading cause of blindness with at least 37 million people infected and more than 120 million people at risk of contracting the disease; most (99%) of this population, threatened by infection, live in Africa. The drug of choice for mass treatment is the microfilaricidal Mectizan(®) (ivermectin); it does not kill the adult stages of the parasite at the standard dose which is a single annual dose aimed at disease control. However, multiple treatments a year with ivermectin have effects on adult worms. The discovery of new therapeutic targets and drugs directed towards the killing of the adult parasites are thus urgently needed. The chitinase of filarial nematodes is a new drug target due to its essential function in the metabolism and molting of the parasite. Closantel is a potent and specific inhibitor of chitinase of Onchocerca volvulus (OvCHT1) and other filarial chitinases. However, the binding mode and specificity of closantel towards OvCHT1 remain unknown. In the absence of a crystallographic structure of OvCHT1, we developed a homology model of OvCHT1 using the currently available X-ray structures of human chitinases as templates. Energy minimization and molecular dynamics (MD) simulation of the model led to a high quality of 3D structure of OvCHIT1. A flexible docking study using closantel as the ligand on the binding site of OvCHIT1 and human chitinases was performed and demonstrated the differences in the closantel binding mode between OvCHIT1 and human chitinase. Furthermore, molecular dynamics simulations and free-energy calculation were employed to determine and compare the detailed binding mode of closantel with OvCHT1 and the structure of human chitinase. This comparative study allowed identification of structural features and properties responsible for differences in the computationally predicted closantel binding modes. The homology model and the closantel binding mode reported herein might help guide the rational development of novel drugs against the adult parasite of O. volvulus and such findings could be extrapolated to other filarial neglected diseases.

Evidence for suppression of Onchocerca volvulus transmission in the Oaxaca focus in Mexico.

Entomologic and serologic surveys were performed in four sentinel communities in the Oaxaca focus in southern Mexico to assess the level of transmission and exposure incidence to Onchocerca volvulus. All communities have been receiving ivermectin mass treatment twice per year since 1997. In one community, parasite DNA was detected by polymerase chain reaction-enzyme-linked immunosorbent assay in 2004 in one pool of 50 vector heads of 170 such pools (8,500 flies) examined, which indicated an estimated transmission potential of 6.7 third-stage larvae/person/year. No evidence for transmission was found in the three other communities in 13,650 flies examined. All persons in a cohort consisting of 117 children in the four communities remained serologically negative for antibodies recognizing a cocktail of recombinant antigens over a four-year period from 2001 to 2004, which indicated an exposure incidence of 0%. Taken together, these data suggest that transmission has been suppressed in the four communities.

Analysis of genetic variation in ribosomal DNA internal transcribed spacer and the NADH dehydrogenase subunit 4 mitochondrial genes of the onchocerciasis vector Simulium ochraceum.

Onchocerciasis is a serious disease vectored by black flies in the genus Simulium that are infected with the filarial parasite Onchocerca volvulus. In the Americas, black flies of the Simulium ochraceum s.l. species complex are important vectors of this parasite. Cytological studies have suggested that this species complex consists of at least three cytotypes that inhabit distinct habitats. In this study, the NADH dehydrogenase subunit four (ND4) and internal transcribed spacer (ITS) of the ribosomal RNA gene cluster were used to explore the degree of genetic diversity among S. ochraceum s.l. populations found in the three O. volvulus foci in Mexico. Both sequence regions were found to exhibit intra- and interpopulation variation. Four different ND4 alleles were found among the populations examined. Similarly, variation was noted in the ITS domain sequences within and among populations. Variation within the ITS sequence was primarily confined to a complex microsatellite locus. Four ITS length variants were observed, two of which were only seen in flies collected from the onchocerciasis focus in northern Chiapas. These data suggest that the ND4 and ITS sequences may prove to be useful markers for exploring interactions within and among the S. ochraceum s.l. populations in Mexico.

Polymerase chain reaction monitoring of transmission of Onchocerca volvulus in two endemic states in Mexico.

To investigate the impact of mass ivermectin treatments in Mexico on Onchocerca volvulus transmission, entomologic surveys were carried out in the two endemic states of Oaxaca and Chiapas. The data suggest that substantial progress towards the goal of elimination has been made. A comparison pre- and post-ivermectin data from a community in Southern Chiapas showed a 97% decrease in seasonal transmission potential, but some level of polymerase chain reaction positivity was still detectable. In other communities from northern Chiapas and Oaxaca where there are no baseline data, there was an absence or near absence of infective flies. Residual transmission was not evenly distributed because differences were seen in the infection and infective rates from different catch points. These findings suggest that while substantial progress towards elimination has been made in Mexico, it may be necessary to modify ivermectin distribution strategies to eliminate transmission in areas where transmission persists.

Rapid suppression of Onchocerca volvulus transmission in two communities of the Southern Chiapas focus, Mexico, achieved by quarterly treatments with Mectizan.

The impact of quarterly Mectizan (ivermectin) treatments on transmission, microfiladermia, and ocular lesions was evaluated in two formerly hyperendemic communities (Las Golondrinas and Las Nubes II) located in the main endemic focus for onchocerciasis in Southern Chiapas, Mexico. The data suggest that Onchocerca volvulus transmission has been suppressed after elimination of microfiladermia in these two communities. Increasing the frequency of Mectizan treatment to four times per year appears to have resulted in the rapid suppression of transmission in communities with residual transmission.