METHODS: By using a multiscale approach, we evaluated the effect of increasing concentrations of MPA on intestinal epithelial cells (Caco-2 cell line) viability, proliferation, and migration. Then, we investigated the inhibitory properties of amoxapine, a previously described bacterial β-G inhibitor, by using molecular dynamics simulations, and evaluated its efficiency in blocking MPAG hydrolysis in an Escherichia coli-based β-G activity assay. The pharmacological effect of amoxapine was evaluated in a mouse model.
RESULTS: We observed that MPA impairs intestinal epithelial cell homeostasis. Amoxapine efficiently blocks the hydrolysis of MPAG to MPA and significantly reduces digestive exposure to MPA in mice. As a result, administration of amoxapine in MPA-treated mice significantly attenuated gastrointestinal lesions.
CONCLUSIONS: Collectively, these results suggest that the digestive accumulation of MPA is involved in the pathophysiology of MPA-gastrointestinal adverse effects. This study provides a proof-of-concept of the therapeutic potential of bacterial β-G inhibitors in glucuronidated drug-induced enteropathy.
方法:通过使用多尺度方法,我们评估了增加浓度的MPA对肠上皮细胞(Caco-2细胞系)活力的影响,扩散,和移民。然后,我们研究了阿莫沙平的抑制特性,先前描述的细菌β-G抑制剂,通过使用分子动力学模拟,并在基于大肠杆菌的β-G活性测定中评估了其阻断MPAG水解的效率。在小鼠模型中评价阿莫沙平的药理作用。
结果:我们观察到MPA损害肠上皮细胞稳态。阿莫沙平可有效阻断MPAG水解为MPA,并显着减少小鼠对MPA的消化暴露。因此,在MPA治疗的小鼠中施用阿莫沙平可显着减轻胃肠道病变。
结论:总的来说,这些结果表明,MPA的消化积累参与了MPA胃肠道不良反应的病理生理过程。这项研究提供了细菌β-G抑制剂在葡萄糖醛酸化药物诱导的肠病中的治疗潜力的概念验证。