ABSTRACT
Mirabegron is a selective beta3-adrenoceptor (ß3 -AR) agonist, which is commonly used for the treatment of overactive bladder. This medicine is associated with atrophy of reproductive organs in rats. However, no study has examined the detailed action and mechanism of its toxicity in reproductive cells. In this study, we examined the effect of mirabegron on primary cultured rat Sertoli cells. Firstly, RT-PCR and immunocytochemistry revealed that ß3 -AR was present in rat Sertoli cells. Then, primary cultured rat Sertoli cells were treated with mirabegron. Quantitative real-time PCR revealed that mirabegron treatment induced a significant increase in claudin-11 mRNA, which is crucial for spermatogenesis. Western blot analysis also showed that mirabegron treatment significantly activated p44/42 mitogen-activated protein kinase (MAPK). After additional treatment with U0126, a specific noncompetitive inhibitor of mitogen-activated protein kinase kinase (MAPKK), the upregulation of claudin-11 mRNA induced by mirabegron was reduced. At the same time, immunocytochemistry showed mirabegron treatment disturbed claudin-11 localisation to tight junction, which was recovered when treated with mirabegron in the presence of U0126. These results suggest that mirabegron treatment is associated with assembly of the blood-testis barrier through p44/42 MAPK pathway. These findings could explain one of the underlying mechanisms of reproductive toxicity induced by mirabegron.
Subject(s)
Acetanilides/toxicity , Adrenergic beta-3 Receptor Agonists/toxicity , Blood-Testis Barrier/drug effects , Sertoli Cells/drug effects , Thiazoles/toxicity , Tight Junctions/drug effects , Animals , Butadienes/pharmacology , Cells, Cultured , Claudins/metabolism , Male , Mitogen-Activated Protein Kinase 1/antagonists & inhibitors , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/antagonists & inhibitors , Mitogen-Activated Protein Kinase 3/metabolism , Nitriles/pharmacology , Primary Cell Culture , Rats , Rats, Sprague-Dawley , Sertoli Cells/cytology , Sertoli Cells/metabolism , Signal Transduction/drug effects , Spermatogenesis/drug effects , Tight Junctions/metabolismABSTRACT
In this study, the frog embryo teratogenesis assay (FETAX - Xenopus) technique was employed to evaluate the potential teratogenicity of the selective ß-adrenoceptor (AR) agonist, CL 316.243. In this context, CL 316.243 was applied to the South African clawed frog (Xenopus laevis) embryos. The media containing the CL 316.24-exposed embryos were monitored and changed/replaced once every 24 hours. Using FETAX, we determined the minimum concentrations to inhibit growth (MCIG) for CL 316.243. The 96-hour no observable adverse effect concentration (NOAEC), the 96-hour lowest observable adverse effect concentration (LOAEC), the 96-hour EC50 (malformation) and the 96-hour LC50 (lethal concentration) for mortality and malformation could not be determined because the used concentrations did not affect viability or the presence of abnormalities. On the other hand, the MCIG of CL 316.243 was determined as 1 mg/L. Our results demonstrated that CL 316.243 administration was associated with no of teratogenic and toxic effects. However, from first concentration we used (1 to 5 mg/L) length of embryos reduced significantly (p < 0.001) when compared to control of Xenopus embryos. Further studies should be conducted with different concentrations in order to investigate the optimal concentrations for treating preterm labor with these substances.
Subject(s)
Adrenergic beta-3 Receptor Agonists/toxicity , Dioxoles/toxicity , Embryo, Nonmammalian/drug effects , Embryonic Development/drug effects , Teratogenesis , Animals , Dose-Response Relationship, Drug , Lethal Dose 50 , No-Observed-Adverse-Effect Level , Toxicity Tests , Xenopus laevisABSTRACT
The discovery of vibegron, a potent and selective human ß3-AR agonist for the treatment of overactive bladder (OAB), is described. An early-generation clinical ß3-AR agonist MK-0634 (3) exhibited efficacy in humans for the treatment of OAB, but development was discontinued due to unacceptable structure-based toxicity in preclinical species. Optimization of a series of second-generation pyrrolidine-derived ß3-AR agonists included reducing the risk for phospholipidosis, the risk of formation of disproportionate human metabolites, and the risk of formation of high levels of circulating metabolites in preclinical species. These efforts resulted in the discovery of vibegron, which possesses improved druglike properties and an overall superior preclinical profile compared to MK-0634. Structure-activity relationships leading to the discovery of vibegron and a summary of its preclinical profile are described.
