NES保有蛋白の核外移行阻害を標的とする新規医薬リード化合物の探索

细胞核输出信号（nuclear export signal， NES）是氨基酸的一个特征序列，近年来，一些具有NES受体蛋白CRM1的蛋白质被证明可以通过细胞核输出到细胞质中。有丝分裂原活化蛋白激酶激酶（MAPKK）和Rev是具有代表性的具有蛋白质的NES，从细胞核输出后在细胞质中发挥重要作用。前者与各种肿瘤细胞的增殖有关，后者是HIV - 1病毒复制所必需的。此外，抑制NES受体CRM1可有效抑制肿瘤细胞的增殖和hiv病毒的复制。迄今为止，leptomycin B(2)和callystatin A(3)仅被证实是CRM1的抑制剂。在此背景下，我们构建了一个检测系统，利用NLS GFP NES转化酵母寻找另一种NES抑制剂，并通过生物检测引导分离从缬草中发现了一种新的NES抑制剂vartrate(1)。通过使用从callystatin A(3)衍生的生物素化探针，我们推测varflate(1)通过与CRM1中的Cys 529结合，以与2和3相同的方式抑制具有蛋白质的NES的输出。此外，1与CRM1的结合位点为1和N乙酰半胱氨酸甲酯的一个环氧基团

Recently, some proteins with nuclear export signal (NES), which is a characteristic sequence of amino acids, were shown to be exported from nucleus to cytoplasm with the aid of NES receptor protein CRM1. Mitogenactivated protein kinase kinase (MAPKK) and Rev were shown to be representative NES possessing proteins and play important roles in cytoplasm after export from nucleus. The former is concerned with proliferation of various tumor cells, the latter is required by replication of HIV 1 virus. In addition, inhibition of NES receptor, CRM1, led to suppress both proliferation of the tumor cells and replication of HIV1 virus potently. Up to date, leptomycin B (2) and callystatin A (3) have been only clarified to be inhibitors of CRM1. In this context, we constructed an assay system to search for another NES inhibitor by using the NLS GFP NES transformed yeast and disclosed a new NES inhibitor, vartrate (1), from Valerianae Radix through bioassayguided separation. By using the biotinylated probe derived from callystatin A (3), varflate (1) was presumed to inhibit export of the NES possessing proteins through binding to the Cys 529 in CRM1 in the same fashion as 2 and 3. Furtherriiore, the binding site of 1 to CRM1 was elucidated to be an epoxy moiety from the reactant of 1 and N acetylcysteine methyl ester