The potential endocrine disruption mechanism of anthelmintic drug niclosamide by activating estrogen receptors and estrogen-related receptors.

Niclosamide (NIC), a helminthic drug used widely for controlling schistosomiasis, can reportedly disrupt the endocrine system. However, its underlying mechanisms are still unclear. In this study, we revealed the potential endocrine disruption mechanism of NIC by activating estrogen receptors (ERs) and estrogen-related receptors (ERRs). The binding potency of NIC with ERα, ERß and ERRγ were determined by fluorescence competitive binding assays, which shows an IC50 (the concentration of NIC needed to displace 50 % of the probe from the receptor) of 90 ± 4.1, 10 ± 1.7 nM and 0.59 ± 0.07 nM respectively. The IC50 for ERRγ is the lowest one among the three detected receptors, which is three orders of magnitude lower than the known agonist GSK4716.The transcriptional activities of NIC on ERs and ERRs were detected by MVLN cells (stably transfected with ERs reporter gene) and HeLa cells (transiently transfected with ERRs reporter gene)-based luciferase reporter gene assay. The lowest observable effective concentration (LOEC) ranked as follows: ERRγ (0.5 nM) < ERRα (10 nM) < ERs (100 nM). The maximum observed induction rate for ERRγ (294 %) was higher than that for ERRα (191 %). The maximum observed induction rate of NIC for ERs was 30 % relative to 17ß-estradiol. In addition, we simulated the interactions of NIC with ERs and ERRs by molecular docking. NIC could dock into the ligand binding pockets of ERs and ERRs and form hydrogen bonds with different amino acids. The binding energy ranked as follows: ERRγ (-8.90 kcal/mol) < ERß (-7.57 kcal/mol) < ERRα (-7.15 kcal/mol) < ERα (-6.53 kcal/mol), which implied that NIC bound to ERRγ with higher binding affinity than the other receptors. Overall, we clarify that ERRγ might be the dominant target for NIC in cells rather than ERRα and ERs. We reveal potential novel mechanisms for the endocrine disruption effects of NIC by activating both ERRs and ERs at environmentally-related nanomolar levels.

Formulation and evaluation of cyclodextrin complexes for improved anticancer activity of repurposed drug: Niclosamide.

Niclosamide, previously used as an anthelmintic drug is currently being repurposed for its anticancer activity. Niclosamide is a brick like biopharmaceutical classification system (BCS) class II drug with poor aqueous solubility and dissolution consequently leading to low bioavailability. By considering the physicochemical properties and geometry of niclosamide, inclusion complex with cyclodextrin was prepared by freeze drying method and characterized using FT-IR, DSC, PXRD, and 1HNMR. In silico molecular modeling study was performed to study the possible interactions between niclosamide and cyclodextrin. The anticancer activity of niclosamide formulation was evaluated through in vitro cell cytotoxicity study using various cancer cell lines. The potential of niclosamide complex for improvement of the bioavailability was evaluated in male BALB/c mice. In vitro cytotoxicity studies indicated significantly higher cytotoxicity at lower concentrations and the pharmacokinetic studies showed significant improvement in Cmax and Tmax of niclosamide from cyclodextrin complex in comparison to pure niclosamide alone.

Scolicidal effects of biogenic selenium nanoparticles against protoscolices of hydatid cysts.

Cystic echinococcosis (hydatid cyst, CE) as a zoonotic parasitic infection caused by the larval stage of the dog tapeworm Echinococcus granulosus is still an important economic and public health concern in the world. One of the treatment options for CE is surgical removal of the cysts combined with chemotherapy using albendazole and/or mebendazole before and after surgery. Currently, many scolicidal agents, which have some complications, have been used for inactivation of the cyst contents. Therefore the development of new scolicidal agents with low side effects and more efficacies is an urgent need for surgeons. The present study was aimed to investigate the in vitro scolicidal effect of selenium nanoparticles biosynthesized by a newly isolated marine bacterial strain Bacillus sp. MSh-1 against protoscoleces of E. granulosus. Protoscolices were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations (50-500 µg/ml) of Se NPs (in size range of about 80-220 nm) were used for 10-60 min. Viability of protoscoleces was confirmed by 0.1% eosin staining. The results indicated that biogenic Se NPs at all concentrations have potent scolicidal effects especially at concentrations 500 and 250 µg/ml after 10 and 20 min of application, respectively. In conclusion, the findings of present study proven that Se NPs have potent scolicidal effects, therefore may be used in CE surgery. However, the in vivo efficacy of these NPs remains to be explored.

Anticestodal activity of endophytic Pestalotiopsis sp. on protoscoleces of hydatid cyst Echinococcus granulosus.

Surgery is still the main treatment in hydatidosis caused by Echinococcus, which is a global health problem in human and animals. So, there is need for some natural protoscolicidal agents for instillation to prevent their reoccurrence at therapeutic doses. In this present investigation, anticestodal activity of one of the endophytic fungi Pestalotiopsis sp. from Neem plant was observed on protoscoleces of hydatid cysts of Echinococcus granulosus. Viability of protoscoleces was confirmed by 0.1% aqueous eosin red stain method, where mortality was observed at different concentrations with respect to time. An average anticestodal activity was observed with different endophytic fungal strains, that is, Nigrospora (479 ± 2.9), Colletotrichum (469 ± 25.8), Fusarium (355 ± 14.5), and Chaetomium (332 ± 28.3) showing 64 to 70% protoscolicidal activity, except Pestalotiopsis sp. (581 ± 15.0), which showed promising scolicidal activity up to 97% mortality just within 30 min of incubation. These species showed significant reduction in viability of protoscoleces. This is the first report on the scolicidal activity of endophytic Pestalotiopsis sp. We conclude that ultrastructural changes in protoscoleces were due to endophytic extract suggesting that there may be some bioactive compounds that have selective action on the tegument layer of protoscoleces. As compared with that of standard drug used, endophytic species of Neem plant shows significant anticestodal activity.

Tissue distribution and excretion of CDRI-81/470 in rats.

Methyl-N[5 [[4-(2-pyridinyl)-1-piperazinyl]carbonyl]- 1H-benzimidazol-2-yl] carbamate (CDRI-81/470) is a broad spectrum anthelmintic agent, effective against both intestinal and systemic parasitism. Tissue distribution and excretion of CDR1-81/470 were studied in rats after a single oral dose of 100 mg kg(-1) CDRI-81/470. One of the metabolites was identified in pilot studies as its N-decarboxylate derivative and characterized by synthesis. HPLC assay methods for the simultaneous estimation of CDRI-81/470 and its N-decarboxylate derivative in tissues, bile, urine, and faeces were developed and validated. The parent compound was quantitated in all major tissues and organs up to 48 h post-dose. Among the tissues other than serum, the highest concentrations of CDRI-81/470 were found in liver, whereas only trace levels were found in brain. Approximately 3% of the administered dose was excreted unchanged in urine at 120 h postdose, whereas approximately 7% was recovered in faeces. The contribution of the biliary route for the excretion of parent compound was less than 0.5%. The N-decarboxylate derivative was quantitated in faeces (1-4%) and bile ( < 0.1%) but was absent in serum, tissues, and urine. An additional metabolite was isolated from bile and characterized as the pyridinyl-5-hydroxy derivative of CDRI-81/470. CDRI-81/470 showed rapid absorption and distribution into all major organs and tissues, and underwent extensive metabolism in rats. Two metabolites in bile were identified and characterized by synthesis.

[Treatment of alveolar echinococcosis with flubendazole. Pharmacological study (author's transl)]. / Traitement de l'échinococcose alvéolaire du foie par le flubendazole. Etude pharmacologique.

Several publications mention a relative efficacy of flubendazole in the treatment of alveolar echinococcosis of the liver (AE). It may seem surprising, being considered the poor intestinal absorption of this drug. A study of the plasma levels was performed in 6 patients, during the treatment with oral flubendazole (approximatively 50 mg/kg/24 h) : 3 patients with normal or subnormal liver functions, 1 hydatidosis and 2 clinically nearly latent AE (group I) ; 3 patients with a highly developed AE (group II). The levels were measured by radioimmunoassay. In the group I, the plasma levels were very low, more often inferior to 3 ng/ml, without distinct peaks. In the group II, the valley levels were higher than 0 ng/ml, and the peaks ranged from 47,4 to 50,4 ng/ml (1 to 3 hours after an intake). The increase in the dosage allowed higher levels, as opposed to previous assertions. The bile and faeces concentrations, as well as the urinary elimination, were studied in a patient with external biliary drainage. The intramusculary injectable flubendazole was used in a patient, without benefit as far as plasma levels are concerned.

[Pharmacokinetic study of flubendazole in human hydatid disease caused by Echinococcus granulosus. Preliminary results (author's transl)]. / Etude pharmacocinétique du flubendazole au cours de l'hydatidose humaine à E. granulosus. Résultats préliminaires.

A pharmacokinetic study of flubendazole was carried out, using a radioimmunological method, in patients with hydatid disease about to undergo surgery; there was no evidence of drug penetration into the walls or contents of the cysts. Another study, carried out simultaneously in subjects without hydatid disease, showed very high concentrations of the drug in liver tissue and in bile. These results should not be taken as arguments against the effectiveness of flubendazole, but should encourage the setting up of further randomized pharmacokinetic studies.