Antischistosomal Drug Praziquantel Research Articles

Ameliorative Effect of Dandelion Leaves Extract Against Liver Injury Induced by Schistosomiasis in Mice

The present study is planned to investigate the effect of dandelion leaves water extract (DWE) on Schistosomiasis. Thirty-five adult male albino mice weighing 20-25 g were used and divided into two main groups, the main first group served as a normal control group without infection (n=7); the second main group was infected with S. mansoni (n=28). After 45 days post-infection the second main group was divided into four subgroups; the first subgroup infected with S. mansoni without treatment (IC) and served as positive control group, the second subgroup infected with S. mansoni then treated by praziquantel (PZQ), the third subgroup infected with S. mansoni then treated by DWE, and the fourth subgroup infected with S. mansoni then treated by DWE in combination with PZQ.The results indicated that administration of combined DWE+PZQ is more effective in all parameters than PZQ or DWE and exhibited highly significantly reduced parasitological parameters, the number and diameter of hepatic granulomas, and histopathological changes in the liver as compared to the untreated infected group. The results also showed that the administration of DWE+PZQ remarkably reduced AST and ALT activities and increment of serum albumin. These amelioration effects were observed in DWE+PZQ treated group, then PZQ‑treated group, followed by the group treated with DWE only. This study concluded that dandelion leaves water extract can be added as a potential treatment against schistosomiasis when combined with the anti-schistosomal drug praziquantel.

CaMKII regulates neuromuscular activity and survival of the human blood fluke Schistosoma mansoni

Calcium/calmodulin dependant protein kinase II (CaMKII), an important transducer of Ca2+ signals, orchestrates multiple cellular functions in animals. Here we investigated the importance of CaMKII to Schistosoma mansoni, a blood parasite that causes human schistosomiasis. We demonstrate that phosphorylated (activated) CaMKII is present in cercariae, schistosomula and adult worms, and show that striking activation occurs in the nervous tissue of these parasite life-stages; CaMKII was also activated in the tegument and muscles of adult worms and the vitellaria of females. Exposure of worms to the anti-schistosomal drug praziquantel (PZQ) induced significant CaMKII activation and depletion of CaMKII protein/activation in adult worms resulted in hypokinesia, reduced vitality and death. At medium confidence (global score ≥ 0.40), S. mansoni CaMKII was predicted to interact with 51 proteins, with many containing CaMKII phosphorylation sites and nine mapped to phosphoproteome data including sites within a ryanodine receptor. The CaMKII network was functionally enriched with mitogen-activated protein kinase, Wnt, and notch pathways, and ion-transport and voltage-dependent channel protein domains. Collectively, these data highlight the intricacies of CaMKII signalling in S. mansoni, show CaMKII to be an active player in the PZQ-mediated response of schistosomes and highlight CaMKII as a possible target for the development of novel anti-schistosome therapeutics.

Itraconazole, a cytochrome P450 inhibitor, enhanced the efficacy of praziquantel against Schistosoma mansoni infection and alleviated liver injury in mice

Treatment of schistosomiasis is heavily reliant on the single antischistosomal drug praziquantel (PZQ). The use of synergistic drug-drug interactions is one possible solution, which could be used to mitigate PZQ's poor and variable bioavailability. Itraconazole (ITZ), a triazole antifungal agent, is a potent CYP3A inhibitor that can cause significant drug-drug interactions when used with CYP3A substrates. This study investigates the effect of ITZ as adjuvant therapy with PZQ on worm load, egg deposition and maturation, and the consequent histopathology and biochemical abnormalities in the liver during the immature and mature stages of Schistosoma mansoni (S. mansoni) infection. S. mansoni-infected mice were divided into five groups of eight−ten mice each: (I) infected untreated, (II) infected and treated with PZQ 3 weeks PI, (III) infected and treated with both ITZ and PZQ 3 weeks PI, (IV) infected and treated with PZQ 7 weeks PI, and (V) infected and treated with both ITZ and PZQ 7 weeks PI. All mice were killed by rapid decapitation 9 weeks PI. Data revealed that ITZ in combination with PZQ at both immature and mature stages improved the parasitological criteria of cure, and greatly reduced inflammation, granuloma and fibrotic tissue formation, and apoptosis versus PZQ alone. Furthermore, it showed the greatest impact on improving liver injury and oxidative stress markers. Notably, the effect was considerably stronger at the mature stage of S. mansoni infection. These findings support the notion that ITZ increased PZQ's antischistosomal activity by inhibiting CYP450 expression, potentially reducing PZQ metabolism and increasing systemic exposure.

Efficacy of carvacryl acetate in vitro and following oral administration to mice harboring either prepatent or patent Schistosoma mansoni infections.

Schistosomiasis is a major public health problem that afflicts more than 240 million individuals globally, particularly in poor communities. Treatment of schistosomiasis relies heavily on a single oral drug, praziquantel, and there is interest in the search for new antischistosomal drugs. This study reports the anthelmintic evaluation of carvacryl acetate, a derivative of the terpene carvacrol, against Schistosoma mansoni ex vivo and in a schistosomiasis animal model harboring either adult (patent infection) or juvenile (prepatent infection) parasites. For comparison, data obtained with gold standard antischistosomal drug praziquantel are also presented. Initially in vitro effective concentrations of 50% (EC50) and 90% (EC90) were determined against larval and adult stages of S. mansoni. In an animal with patent infection, a single oral dose of carvacryl acetate (100, 200, or 400mg/kg) caused a significant reduction in worm burden (30-40%). S. mansoni egg production, a process responsible for both life cycle and pathogenesis, was also markedly reduced (70-80%). Similar to praziquantel, carvacryl acetate 400mg/kg had low efficacy in pre-patent infection. In tandem, although carvacryl acetate had interesting in vitro schistosomicidal activity, the compound exhibited low efficacy in terms of reduction of worm load in S. mansoni-infected mice.

Schistosomiasis.

Schistosomiasis (bilharzia) is a neglected tropical disease caused by parasitic flatworms (blood flukes) of the genus Schistosoma, with considerable morbidity in parts of the Middle East, South America, Southeast Asia and, particularly, in sub-Saharan Africa. Infective larvae grow in an intermediate host (fresh-water snails) before penetrating the skin of the definitive human host. Mature adult worms reside in the mesenteric (Schistosoma mansoni and Schistosoma japonicum) or pelvic (Schistosoma haematobium) veins, where female worms lay eggs, which are secreted in stool or urine. Eggs trapped in the surrounding tissues and organs, such as the liver and bladder, cause inflammatory immune responses (including granulomas) that result in intestinal, hepato-splenic or urogenital disease. Diagnosis requires the detection of eggs in excreta or worm antigens in the serum, and sensitive, rapid, point-of-care tests for populations living in endemic areas are needed. The anti-schistosomal drug praziquantel is safe and efficacious against adult worms of all the six Schistosoma spp. infecting humans; however, it does not prevent reinfection and the emergence of drug resistance is a concern. Schistosomiasis elimination will require a multifaceted approach, including: treatment; snail control; information, education and communication; improved water, sanitation and hygiene; accurate diagnostics; and surveillance-response systems that are readily tailored to social-ecological settings.

Praziquantel Interaction with Mammalian Targets in the Spotlight

Chan et al. recently demonstrated that the antischistosomal drug praziquantel has a potent and specific interaction with human 5-HT2B receptors, and that the drug also elicits contraction of mouse mesenteric vasculature apparently mediated by the same receptor subtype We consider what this might mean about the drug's molecular therapeutic targets in both the worm and the host.

A palladium-catalyzed intramolecular Heck/Hydrogenation approach towards the synthesis of praziquantel

Starting from 3-methoxy N-acylpyrazinium salts, a new approach towards the synthesis of the antischistosomal drug praziquantel (PZQ) has been developed. Utilization of a palladium-catalyzed intramolecular Heck reaction to form dehydro-PZQ followed by a Hydrogenation step, in a stepwise or one-pot manner, allowed for the gram scale synthesis of PZQ in 4 or 5 steps and in good overall yields. This methodology proved to be well suited for generating the pyrazino[1,2-a] isoindole and pyrazino[1,2-a] benzazepine analogues of PZQ.

The roles of water, sanitation and hygiene in reducing schistosomiasis: a review.

Schistosomiasis is a disease caused by infection with blood flukes of the genus Schistosoma. Transmission of, and exposure to, the parasite result from faecal or urinary contamination of freshwater containing intermediate host snails, and dermal contact with the same water. The World Health Assembly resolution 65.21 from May 2012 urges member states to eliminate schistosomiasis through preventive chemotherapy (i.e. periodic large-scale administration of the antischistosomal drug praziquantel to school-aged children and other high-risk groups), provision of water, sanitation and hygiene (WASH) and snail control. However, control measures focus almost exclusively on preventive chemotherapy, while only few studies made an attempt to determine the impact of upgraded access to safe water, adequate sanitation and good hygiene on schistosome transmission. We recently completed a systematic review and meta-analysis pertaining to WASH and schistosomiasis and found that people with safe water and adequate sanitation have significantly lower odds of a Schistosoma infection. Importantly though, the transmission of schistosomiasis is deeply entrenched in social-ecological systems, and hence is governed by setting-specific cultural and environmental factors that determine human behaviour and snail populations. Here, we provide a comprehensive review of the literature, which explores the transmission routes of schistosomes, particularly focussing on how these might be disrupted with WASH-related technologies and human behaviour. Additionally, future research directions in this area are highlighted.

The potential role of Morus alba leaves extract on the brain of mice infected with Schistosoma mansoni.

Schistosomiasis is a neglected tropical disease which is associated with neuropsychiatric and neuropathological disorders. Herein, the main goal of the presented work is to investigate the effect of Morus alba leaves extract in mice brain infected with Schistosoma mansoni. Since, the resistance of Schistosomes to antischistosomal drug (praziquantel) has been examined, schistosomiasis induced brain oxidative stress as evidenced by the decrease of glutathione level, total antioxidant capacity and the activity of catalase significantly, while a significant elevation in the levels of nitrite/nitrate and malondialdhyde. In addition, the infection resulted in neurochemical disturbances, the main inhibitory amino acid, γ- aminobutyric acid level was decreased. In contrast, the level of chloride ions and acetylcholine esterase activity were significantly increased. Moreover, the histopathological section showed some impairments in the brain. The treatment with Morus alba leaves extract ameliorated the induced disturbances in schistosome-infected mice where the levels of non-enzymatic and enzymatic antioxidants were elevated. On the other hand, the levels of nitrite/nitrate and malondialdhyde were significantly reduced. Likewise, treatment of mice with Morus alba leaves extract improved the altered levels of γ- aminobutyric acid level and chloride ion. Also, it improved the recorded impairments of the histopathological section in the brain of schistosome infected mice.

In Vitro Metabolic Profile and in Vivo Antischistosomal Activity Studies of (η6-Praziquantel)Cr(CO)3 Derivatives

In vitro metabolic behavior was investigated for two chromium tricarbonyl derivatives of the antischistosomal drug praziquantel (PZQ) with the formula (η(6)-PZQ)Cr(CO)3 (1 and 2), by use of human liver microsomes. The metabolic profiles of the derivatives differ significantly. The optically pure (η(6)-PZQ)Cr(CO)3 derivatives (S, Sp)-1, (R, Rp)-1, (S, Rp)-2, and (R, Sp)-2 were also prepared to assess the eudysmic ratios of 1 and 2 against Schistosoma mansoni in vitro. A strong enantioselective antischistosomal activity was observed. The R-enantiomers are highly active against adult schistosomes in vitro (IC50 0.08-0.13 μM), whereas both S-enantiomers lack activity. The in vivo activity of 1 and 2 was then studied in mice harboring a chronic S. mansoni infection. A single dose of 1 and 2 (400 mg/kg) resulted in low worm burden reductions of 24% and 29% (p > 0.05).

Ca2 + channels and praziquantel: A view from the free world

Targeting the cellular Ca2+ channels and pumps that underpin parasite Ca2+ homeostasis may realize novel antihelmintic agents. Indeed, the antischistosomal drug praziquantel (PZQ) is a key clinical agent that has been proposed to work in this manner. Heterologous expression data has implicated an action of PZQ on voltage-operated Ca2+ channels, although the relevant in vivo target of this drug has remained undefined over three decades of clinical use. The purpose of this review is to bring new perspective to this issue by discussing the potential utility of free-living planarian flatworms for providing new insight into the mechanism of PZQ action. First, we discuss in vivo functional genetic data from the planarian system that broadly supports the molecular data collected in heterologous systems and the ‘Ca2+ hypothesis’ of PZQ action. On the basis of these similarities we highlight our current knowledge of platyhelminth voltage operated Ca2+ channels, their unique molecular pharmacology and the downstream functional PZQ interactome engaged by dysregulation of Ca2+ influx that has potential to yield novel antischistosomal targets. Overall the broad dataset underscores a common theme of PZQ-evoked disruptions of Ca2+ homeostasis in trematodes, cestodes and turbellarians, and showcases the utility of the planarian model for deriving insight into drug action and targets in parasitic flatworms.

Synthesis of new praziquantel analogues: Potential candidates for the treatment of schistosomiasis

An efficient synthesis of antischistosomal drug praziquantel and analogues was achieved and the synthetic route designed was to afford structurally diverse analogues for better structure-activity relationship understanding. Total of nineteen PZQ analogues with structural variations at amide, piperazine and aromatic moieties have been synthesized and fully characterized. Among all the new analogues tested for antischistosomal activity, one dimethoxy tetrahydroisoquinoline analogue and two tetrahydro-β-carboline analogues exhibited moderate activity against adult Schistosoma mansoni. Tetrahydro-β-carboline analogues showed moderate activity whereas the presence of p-trifluoromethylbenzoyl and p-toluenesulphonyl moieties resulted in complete suppression of antischistosomal activity.

Schistosoma mansoni Changes the Activity of Phase II Drug-Metabolizing Enzymes: Role of Praziquantel as Antibilharzial Drug

Schistosomiasis is one of the major health problems in many developing countries and causes liver damage. In addition, under the influence of schistosomiasis most of the endogenous toxic compounds can be conjugated with glutathione via glutathione S-transferase. Therefore, the present study showed the effect of Schistosoma mansoni after 20, 30, 45, 60, and 75 days post-infection on the activity of glutathione-S-transferase (GST) and glutathione reductase (GR), and on the levels of glutathione [GSH] in the livers of male mice. In addition, anti-schistosomal drug (praziquantel) was administered orally [60 mg/kg body weight] for three consecutive days before decapitation of the infected mice at each time point. In the present, depletion of GSH levels was observed at 45, 60 and 75 days post-infection. However, treatment of infected mice at 45, 60, and 75 days post-infection with praziquantel for three consecutive days before decapitation at each time point restored the increased GSH levels to their normal values compared with control groups. Inhibition of GST and induction of GR activities in the livers of S. mansoni-infected mice at all time-points were restored to their normal levels after praziquantel treatment. It is concluded that S. mansoni infection changed the activities of GST, GR and GSH levels. Moreover, it has been found that praziquantel treatment of S. mansoni-infected mice restored such alterations to their normal values and this recovery could alleviate the deleterious effects of S. mansoni infection. In addition, the present study could provide new evidence to the damage occurred in livers of S. mansoni-infected hosts. Also, it is suggested that praziquantel is the best drug of choice for schistosoma mansoni treatment.

A Novel Biological Activity of Praziquantel Requiring Voltage-Operated Ca2+ Channel β Subunits: Subversion of Flatworm Regenerative Polarity

BackgroundApproximately 200 million people worldwide harbour parasitic flatworm infections that cause schistosomiasis. A single drug—praziquantel (PZQ)—has served as the mainstay pharmacotherapy for schistosome infections since the 1980s. However, the relevant in vivo target(s) of praziquantel remain undefined.Methods and FindingsHere, we provide fresh perspective on the molecular basis of praziquantel efficacy in vivo consequent to the discovery of a remarkable action of PZQ on regeneration in a species of free-living flatworm (Dugesia japonica). Specifically, PZQ caused a robust (100% penetrance) and complete duplication of the entire anterior-posterior axis during flatworm regeneration to yield two-headed organisms with duplicated, integrated central nervous and organ systems. Exploiting this phenotype as a readout for proteins impacting praziquantel efficacy, we demonstrate that PZQ-evoked bipolarity was selectively ablated by in vivo RNAi of voltage-operated calcium channel (VOCC) β subunits, but not by knockdown of a VOCC α subunit. At higher doses of PZQ, knockdown of VOCC β subunits also conferred resistance to PZQ in lethality assays.ConclusionsThis study identifies a new biological activity of the antischistosomal drug praziquantel on regenerative polarity in a species of free-living flatworm. Ablation of the bipolar regenerative phenotype evoked by PZQ via in vivo RNAi of VOCC β subunits provides the first genetic evidence implicating a molecular target crucial for in vivo PZQ activity and supports the ‘VOCC hypothesis’ of PZQ efficacy. Further, in terms of regenerative biology and Ca2+ signaling, these data highlight a novel role for voltage-operated Ca2+ entry in regulating in vivo stem cell differentiation and regenerative patterning.

The anti-schistosomal drug praziquantel is an adenosine antagonist

The mechanism of action of praziquantel (PZQ), the drug of choice against schistosomiasis, is still unclear. Since exposure of schistosomes to the drug is associated with calcium influx and muscular contraction, calcium channels have been suggested as the target, although direct combination of PZQ with their subunits was never demonstrated. We report a hitherto unknown effect of PZQ, namely the inhibition of nucleoside uptake, as observed in living worms using radio-isotope labelled adenosine and uridine. This effect is clearly seen in schistosomes but is absent in mammalian cells in culture. Moreover it is a specific pharmacological effect seen exclusively with the active levo-R(-)stereo isomer of the drug, and is shared by at least one benzodiazepine having antischistosomal activity. This novel effect acquires significance given that schistosomes cannot synthesize purine nucleosides de novo. A possible relationship between this novel effect and the known action of PZQ on calcium channels is discussed, since adenosine is known to bind to specific receptors and to behave as an indirect antagonist of calcium release in mammalian cells. If calcium channels were correlated with adenosine receptors also in schistosomes, as they are in mammals, this would support the hypothesis that PZQ-induced calcium influx may be correlated to adenosine receptor blockade.

Efficiency of Immunization of Mice with Irradiated Antigen Against Schistosoma mansoni Infection in Comparison with Praziquantel

Introduction: The present study is an attempt to evaluate the protective effect of schistosomula antigen and the current antischistosomal drug praziquantel (PZQ) as a reference drug on mice infected with S. mansoni. Material and Methods: Mice were vaccinated by irradiated or non-irradiated schistosomula antigen, both at a dose of 100 ug protein/mice once weekly for 3 weeks, before infection with alive cercariae and compared with the treatment with i.m. injection of praziquantel at a dose of 40 mg/kg b.wt. 4 times once weekly for 4 weeks after infection. The degree of resistance or protection induced by immunization and chemotherapy was assessed 45 days post–infection and evaluated by physiological, parasitological, immunological as well as histological parameters. Results: The results indicated that immunization with γ-irradiated antigen at 20 Krad or the treatment with PZQ resulted in significant reduction in ova count in liver and intestine tissues more than those vaccinated with non-irradiated antigen compared with infected group. Immunized group with irradiated antigen and the group treated with PZQ showed a significant decrease in liver enzymes activity (ALT, AST and γ-GT), while in immunized group with non- irradiated antigen, there was a significant increase in AST and γ-GT as compared to infected group. The level of alkaline phosphatase enzyme was significantly increased in all investigated groups compared to infected one. Treatment with PZQ or immunization with irradiated or non- irradiated schistosomula antigen induced amelioration in serum IL-10 and TNF-α. Scanning electron microscope demonstrated normal mature worms in infected group after 45 days from infection. In contrast, many changes were detected in the rest groups as alterations in the tegument, implosion of tubercles which appeared pealed and sloughed off and most of the spines were detached and separated. Histological examination of liver sections of infected mice revealed lobular cellular infiltration and cloudy swelling in hepatocytes with occurrence of necrotic foci. Also, granuloma of infiltrating cells was noticed around the portal structures and inbetween the degenerated cells. Congested portal vein could be seen lodged with adult worms and the portal tracts showed fibrous thickening. Whereas, the lung revealed destructed bronchioles which appeared surrounded by intense inflammatory foci. Thickened interalveolar septae were also marked, however many alveoli appeared with shed destroyed epithelium. Moreover, peribronchiolar and perivascular fibrosis was quite prominent. Gross pathological alterations were observed in both liver and lung of immunized groups with non-irradiated antigen. In controversy, immunization with irradiated antigen can reduce the granulomatous reaction and collagen deposition. There are also little inflammation and less congestion, however the hepatic and pulmonary architectures appeared otherwise normal. The present histological findings proved that PZQ has a valuable schistosomicidal effect but some pathological changes are still detectable. Conclusion:On the whole, it could be concluded that irradiated antigen produced marked protection against S.mansoni infection and this may reflect its possible beneficial effect on the diseased liver and lung.

Efficiency of Immunization of Mice with Irradiated Antigen Against Schistosoma mansoni Infection in Comparison with Praziquantel

The present study is an attempt to evaluate the protective effect of schistosomula antigen and the current antischistosomal drug praziquantel (PZQ) as a reference drug on mice infected with

Development of antischistosomal drugs in China, with particular consideration to praziquantel and the artemisinins

Remarkable achievements have been made in the control of schistosomiasis in China, with chemotherapy playing a seminal role. From the early 1950s through the early 1980s, Chinese scientists made considerable progress in discovery and development of compounds with antischistosomal properties, including antimonials, non-antimonials and various effective principles stemming from traditional herbs. However, only few compounds entered clinical testing, while others were abandoned mainly due to their toxicity and poor efficacy. The advent of praziquantel in the 1970s changed the landscape of research and development of drugs for treatment and morbidity control of schistosomiasis. The main Chinese contributions to enhance the understanding of the antischistosomal drug praziquantel are reviewed here, including issues of metabolism, antibody-dependency, host immune factors, stage-specific susceptibility and resistance. Over the past 25 years, researchers from China successfully developed artemether and artesunate, two derivatives from the antimalarial artemisinin, as promising drugs against Schistosoma japonicum. Laboratory investigations showed that the artemisinins display their highest activity against the juvenile stages of the parasite. These findings were consistently confirmed in randomised controlled trials; repeated oral administration of artemether or artesunate was safe and efficacious in the prevention of patent S. japonicum infections. The key findings are reviewed here, and emphasis is placed on how it stimulated research outside of China on other human schistosome species.

Specific sites in the Beta Interaction Domain of a schistosome Ca2+ channel beta subunit are key to its role in sensitivity to the anti-schistosomal drug praziquantel.

Praziquantel, the drug of choice against schistosomiasis, disrupts calcium (Ca2+) homeostasis in schistosomes via an unknown mechanism. Voltage-gated Ca2+ channels are heteromultimeric transmembrane protein complexes that contribute to impulse propagation and also regulate intracellular Ca2+ levels. Beta subunits modulate the properties of the pore-forming alpha1 subunit of high voltage-activated Ca2+ channels. Unlike other Ca2+ channel beta subunits, which have current stimulatory effects, a beta subunit subtype found in S. mansoni (SmbetaA) and S. japonicum (Sjbeta) dramatically reduces current levels when co-expressed with Ca2+ channel alpha1 subunits in Xenopus oocytes. It also confers praziquantel sensitivity to the mammalian Cav2.3 alpha1 subunit. The Beta Interaction Domains (BIDs) of SmbetaA and Sjbeta lack 2 conserved serines that each constitute a consensus site for protein kinase C (PKC) phosphorylation. Here, we use site-directed mutagenesis of schistosome beta subunits to show that these unique functional properties are correlated with the absence of these consensus PKC sites in the BID. Furthermore, a second schistosome beta subunit subtype contains both serines in the BID, enhances currents through alpha1 subunits, and does not confer praziquantel sensitivity. Thus, phosphorylation sites in the BID may play important roles in defining the modulatory properties and pharmacological sensitivities of schistosome Ca2+ channel beta subunits.

Creation by mutagenesis of a mammalian Ca 2+ channel β subunit that confers praziquantel sensitivity to a mammalian Ca 2+ channel

Voltage-gated Ca 2+ channel β subunits are important modulators of the pore-forming α 1 subunit. We have cloned two schistosome β subunits that confer sensitivity to the antischistosomal drug praziquantel (PZQ) to an otherwise insensitive mammalian α 1 subunit. The primary site of β subunit interaction with α 1 subunits is the beta interaction domain (BID). The BID contains two conserved serines (225, 235 in rat β2a) that constitute consensus sites for protein kinase C phosphorylation. However, these serines are absent in these schistosome β subunits. Here we show that the capability to confer PZQ sensitivity can be created in the rat β2a subunit by eliminating both serines in the BID. These results are consistent with, and should help our understanding of, the selective toxicity of PZQ.