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目的发现新结构的β-分泌酶抑制剂。方法基于β-分泌酶抑制剂的构效关系,设计合成了含羟乙胺结构片段的间位1,1-二氧代-2,3-(2H)-异噻唑酮和1,1-二氧代-异噻唑啉酮取代的间苯二甲酸衍生物;利用时间分辨荧光法(TRF)和中国仓鼠卵巢细胞(CHO,APP/BACE1基因)模型分别评价这些化合物体外抑制β-分泌酶和Aβ40的活性。结果合成了16个新目标化合物,利用MS和1H-NMR对化合物的结构进行了确证,利用HPLC对化合物的纯度进行了测定,利用旋光仪测定了化合物的比旋光度。活性数据显示,化合物2d、2e、2f、2g、2h和3d、3h抑制β-分泌酶活性的IC50值在10 nmol.L-1以下,化合物2c和3c、3g抑制β-分泌酶活性的IC50值在25 nmol.L-1以下;化合物2c、2d和2h在浓度为0.01μmol.L-1时,对双转APP/BACE1基因CHO细胞Aβ40产生的抑制率分别为58.3%、43.7%和42.1%。结论发现了新的β-分泌酶抑制剂,分析了其初步的构效关系,为进一步的结构优化以及发现活性更好的化合物提供了指导。
Aim To find a novel β-secretase inhibitor. Methods Based on the structure-activity relationship of β-secretase inhibitors, a series of novel 1,1-dioxo-2,3- (2H) -isothiazolones and 1,1-bis Oxo-isothiazolinone-substituted isophthalic acid derivatives were used to evaluate the inhibitory effect of these compounds on β-secretase and Aβ40 in vitro using the time-resolved fluorescence (TRF) and Chinese hamster ovary cells (CHO, APP / BACE1) Activity. Results Sixteen new target compounds were synthesized. The structures of the compounds were confirmed by MS and 1H-NMR. The purity of the compounds was determined by HPLC. The specific rotation of the compounds was determined by polarimetry. The activity data showed that IC50 values of compounds 2d, 2e, 2f, 2g, 2h and 3d, 3h for inhibiting? -secretase activity were below 10 nmol.L-1 and IC50s for compounds 2c and 3c, 3g for inhibiting? -secretase activity The inhibitory rates of Aβ40 in CHO cells transfected with APP / BACE1 double transgenic lines were 58.3%, 43.7% and 42.1% at the concentrations of 0.01μmol.L-1 for compounds 2c, 2d and 2h, respectively %. Conclusions A new β-secretase inhibitor was discovered and its preliminary structure-activity relationship was analyzed, which provided guidance for further structural optimization and discovery of more active compounds.