Abstract：
The readthrough mechanism, which skips the premature termination codon and restores the biosynthesis of the defective enzyme, is an emerging therapeutic tactic for nonsense mutation-related diseases, such as Hurler syndrome, a type of mucopolysaccharidosis. In the present study, novel triaryl derivatives were synthesized and their readthrough-inducing activities were evaluated by a luciferase reporter assay with a partial α-L-iduronidase (IDUA) DNA sequence containing the Q70X nonsense mutation found in Hurler syndrome and by measuring the enzyme activity of IDUA knockout cells transfected with the mutant IDUA gene. KY-516, a representative compound in which the meta position carboxyl group of the left ring of the clinically used ataluren was converted to the para position sulfamoylamino group, the central ring to triazole, and the right ring to cyanobenzene, exhibited the most potent readthrough-inducing activity in the Q70X/luciferase reporter assay. In Q70X mutant IDUA transgenic cells, KY-516 significantly increased enzyme activity at 0.1 µM. After the oral administration of KY-516 (10 mg/kg), the highest plasma concentration of KY-516 was above 5 µM in rats. These results indicate that KY-516, a novel triaryl derivative, exhibits potent readthrough-inducing activity and has potential as a therapeutic agent for Hurler syndrome.
摘要：
通读机制，跳过提前终止密码子，恢复缺陷酶的生物合成，是一种针对无意义突变相关疾病的新兴治疗策略，比如Hurler综合征,粘多糖贮积症的一种.在本研究中,合成了新的三芳基衍生物，并通过荧光素酶报告基因测定法评估了它们的连读诱导活性，该荧光素酶报告基因含有在Hurler综合征中发现的Q70X无义突变的部分α-L-艾杜糖醛酸酶（IDUA）DNA序列，并通过测量用突变IDUA基因转染的IDUA敲除细胞的酶活性。KY-516，一种代表性化合物，其中临床上使用的ataluren的左环的间位羧基被转化为对位氨磺酰氨基，中央环到三唑，和正确的环氰基苯，在Q70X/荧光素酶报告基因测定中表现出最有效的读通诱导活性。在Q70X突变体IDUA转基因细胞中，KY-516在0.1μM时显著增加酶活性。口服KY-516（10mg/kg）后，在大鼠中,KY-516的最高血浆浓度高于5µM.这些结果表明，KY-516，一种新型的三芳基衍生物，具有有效的通读诱导活性，并具有作为Hurler综合征治疗剂的潜力。
