In this study, bufalin was glycosylated by an efficient chemo‐enzymatic strategy. Firstly, 2‐chloro‐4‐nitrophenyl‐1‐O‐β‐D‐glucoside (sugar donors) was obtained by chemical synthesis. Then, the glycosylation of the bufalin was achieved with the synthesized sugar donor under the catalysis of two glycosyltransferases (Loki and ASP). Finally, two glycosides, i. e., bufalin‐3‐O‐β‐D‐glucopyranoside and bufalin‐3‐O‐[β‐D‐glucopyranosyl‐(1→2)‐β‐D‐glucopyranoside)], were obtained by preparative HPLC. Compared to our previously reported sole chemical (total yield 10 % in four steps) or enzymatic methods (30 %), our combined chemo‐enzymatic strategy in this article greatly improves the yields of monoglycoside (68 %) and diglycoside (21 %) and decreased the experimental cost (90 %). Furthermore, we tested the water solubility of these glycosides and found that the water solubilities of the two glycosides were 13.1 and 53.7 times of bufalin, respectively. In addition, the inhibitory activity of these glycosides against Na+, K+‐ATPase were evaluated. The mono‐glycosylated compound showed more potent activity than bufalin, while the diglycosylated compound was less potent.

中文翻译：

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一种化学酶法合成蟾毒甙的有效策略，具有改善的水溶性和对 Na+,K+-ATPase 的抑制作用

在这项研究中，蟾蜍灵被一种有效的化学酶促策略糖基化。首先，通过化学合成获得2-氯-4-硝基苯基-1-O-β-D-葡萄糖苷（糖供体）。然后，在两种糖基转移酶（Loki 和 ASP）的催化下，合成的糖供体实现了蟾蜍灵的糖基化。最后，两个糖苷，i。例如，bufalin-3-O-β-D-吡喃葡萄糖苷和bufalin-3-O-[β-D-吡喃葡萄糖苷-(1→2)-β-D-吡喃葡萄糖苷)]，通过制备型HPLC获得。与我们之前报道的单一化学方法（四步总产率为 10%）或酶法（30%）相比，我们在本文中结合化学酶法的策略大大提高了单糖苷（68%）和双糖苷（21%）的产率和降低了实验成本 (90%)。此外，我们测试了这些苷的水溶性，发现两种苷的水溶性分别是蟾蜍灵的13.1和53.7倍。此外，还评估了这些糖苷对 Na+、K+-ATPase 的抑制活性。单糖基化化合物显示出比蟾蜍灵更强的活性，而二糖基化化合物的活性较低。