Díaz-Herrero M Mar, del Campo José A, Carbonero-Aguilar Pilar, Vega-Pérez José M, Iglesias-Guerra Fernando, Periñán Ignacio, Miñano Francisco J, Bautista Juan, Romero-Gómez Manuel
Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Sevilla, Sevilla, Spain.
Unidad de Gestión Clínica de Enfermedades Digestivas & Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Hospital Universitario de Valme, Sevilla, Spain.
PLoS One. 2014 Oct 17;9(10):e109787. doi: 10.1371/journal.pone.0109787. eCollection 2014.
Ammonia production is implicated in the pathogenesis of hepatic encephalopathy (HE), being intestinal glutaminase activity the main source for ammonia. Management of ammonia formation can be effective in HE treatment by lowering intestinal ammonia production. The use of glutaminase inhibitors represents one way to achieve this goal. In this work, we have performed a search for specific inhibitors that could decrease glutaminase activity by screening two different groups of compounds: i) a group integrated by a diverse, highly pure small molecule compounds derived from thiourea ranging from 200 to 800 Daltons; and ii) a group integrated by commonly use compounds in the treatment of HE. Results shown that THDP-17 (10 µM), a thiourea derivate product, could inhibit the intestinal glutaminase activity (57.4±6.7%). Inhibitory effect was tissue dependent, ranging from 40±5.5% to 80±7.8% in an uncompetitive manner, showing Vmax and Km values of 384.62 µmol min(-1), 13.62 mM with THDP-17 10 µM, respectively. This compound also decreased the glutaminase activity in Caco-2 cell cultures, showing a reduction of ammonia and glutamate production, compared to control cultures. Therefore, the THDP-17 compound could be a good candidate for HE management, by lowering ammonia production.
氨的产生与肝性脑病(HE)的发病机制有关,肠道谷氨酰胺酶活性是氨的主要来源。通过降低肠道氨的产生来控制氨的形成可能对HE治疗有效。使用谷氨酰胺酶抑制剂是实现这一目标的一种方法。在这项研究中,我们通过筛选两组不同的化合物来寻找能够降低谷氨酰胺酶活性的特异性抑制剂:i)一组由多种200至800道尔顿的高纯硫脲衍生小分子化合物组成;ii)一组由HE治疗中常用的化合物组成。结果表明,硫脲衍生物产品THDP-17(10µM)可抑制肠道谷氨酰胺酶活性(57.4±6.7%)。抑制作用具有组织依赖性,以非竞争性方式在40±5.5%至80±7.8%之间,在10µM THDP-17时Vmax和Km值分别为384.62µmol min(-1)、13.62 mM。该化合物还降低了Caco-2细胞培养物中的谷氨酰胺酶活性,与对照培养物相比,氨和谷氨酸的产生减少。因此,THDP-17化合物通过降低氨的产生可能是HE治疗的良好候选药物。