Small molecule inhibitors of intestinal epithelial anion exchanger SLC26A3 (DRA) with a luminal, extracellular site of action.
Small molecule inhibitors of intestinal epithelial anion exchanger SLC26A3 (DRA) with a luminal, extracellular site of action.
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肠上皮阴离子交换剂 SLC26A3 (DRA) 的小分子抑制剂，具有管腔细胞外作用位点。
DOI：
10.1016/j.ejmech.2023.115149
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发表时间：
2023
影响因子：
6.7
通讯作者：
Verkman,AlanS
中科院分区：
文献类型：
--
作者：
Cil,Onur;Anderson,MarcO;deSouzaGoncalves,Livia;Tan,Joseph-Anthony;Haggie,PeterM;Verkman,AlanS
阴离子交换蛋白SLC 26 A3（在腺瘤中下调，腺瘤）在结肠肠上皮细胞的腔膜中表达，在那里它促进Cl−和草酸盐的吸收。我们之前发现了一类4，8-二甲基香豆素SLC 26 A3抑制剂，其从SLC 26 A3细胞质表面起作用，并证明了它们在便秘和高草酸尿症小鼠模型中的功效。在这里，从初步筛选中筛选了50，000种新化合物和1740种活性化合物的化学类似物，产生了5种新型的SLC 26 A3选择性抑制剂（1，3-二氧代异吲哚啉-酰胺; N-（5-氨磺酰基-1，3，4-噻二唑-2-基）乙酰胺;噻唑并嘧啶-5-酮; 3-羧基-2-苯基苯并呋喃和苯并恶嗪-4-酮），IC 50低至100 nM。动力学洗脱和起效研究揭示了噻唑并嘧啶-5-酮和3-羧基-2-苯基苯并呋喃抑制剂的细胞外作用位点。分子对接计算揭示了这些抑制剂的假定结合位点。在小鼠便秘的洛哌丁胺模型中，经口给予7-（2-氯-苯氧基甲基）-3-苯基-噻唑并[3，2-a]嘧啶-5-酮（3a）显著增加粪便重量、颗粒数和含水量。具有细胞外作用位点的SLC 26 A3抑制剂提供了产生不可吸收的、腔道作用抑制剂的可能性，口服后全身暴露量最小。我们的研究结果还表明，抑制剂相关的SLC 26阴离子转运与细胞外的网站的行动可能会被确定为选定的上皮离子转运过程的药理学调制。
The anion exchanger protein SLC26A3 (down-regulated in adenoma, DRA) is expressed in the luminal membrane of intestinal epithelial cells in colon, where it facilitates the absorption of Cl−and oxalate. We previously identified a 4,8-dimethylcoumarin class of SLC26A3 inhibitors that act from the SLC26A3 cytoplasmic surface, and demonstrated their efficacy in mouse models of constipation and hyperoxaluria. Here, screening of 50,000 new compounds and 1740 chemical analogs of active compounds from the primary screen produced five novel classes of SLC26A3-selective inhibitors (1,3-dioxoisoindoline-amides; N-(5-sulfamoyl-1,3,4-thiadiazol-2-yl)acetamides; thiazolo-pyrimidin-5-ones; 3-carboxy-2-phenylbenzofurans and benzoxazin-4-ones) with IC50down to 100 nM. Kinetic washout and onset of action studies revealed an extracellular site of action for the thiazolo-pyrimidin-5-one and 3-carboxy-2-phenylbenzofuran inhibitors. Molecular docking computations revealed putative binding sites for these inhibitors. In a loperamide model of constipation in mice, orally administered 7-(2-chloro-phenoxymethyl)-3-phenyl-thiazolo [3,2-a]pyrimidin-5-one (3a) significantly increased stool weight, pellet number and water content. SLC26A3 inhibitors with an extracellular site of action offer the possibility of creating non-absorbable, luminally acting inhibitors with minimal systemic exposure following oral administration. Our findings also suggest that inhibitors of related SLC26 anion transporters with an extracellular site of action might be identified for pharmacological modulation of selected epithelial ion transport processes.