Prichard Kate, Chau Ngoc, Xue Jing, Krauss Michael, Sakoff Jennette A, Gilbert Jayne, Bahnik Claudia, Muehlbauer Maria, Radetzki Silke, Robinson Phillip J, Haucke Volker, McCluskey Adam
Chemistry, School of Environmental & Life Sciences, The University of Newcastle, University Drive, Callaghan, NSW, 2308, Australia.
Cell Signalling Unit, Children's Medical Research Institute, The University of Sydney, Hawkesbury Road, Westmead, Sydney, Australia.
ChemMedChem. 2024 Oct 16;19(20):e202400253. doi: 10.1002/cmdc.202400253. Epub 2024 Sep 29.
Twenty-five chimera compounds of Pitstop 1 and 2 were synthesised and screened for their ability to block the clathrin terminal domain-amphiphysin protein-protein interaction (NTD-PPI using an ELISA) and clathrin mediated endocytosis (CME) in cells. Library 1 was based on Pitstop 2, but no notable clathrin PPI or in-cell activity was observed. With the Pitstop 1, 16 analogues were produced with 1,8-naphthalic imide core as a foundation. Analogues with methylene spaced linkers and simple amides showed a modest to good range of PPI inhibition (7.6-42.5 μM, naphthyl 39 and 4-nitrophenyl 40 respectively) activity. These data reveal the importance of the naphthalene sulfonate moiety, with no des-SO analogue displaying PPI inhibition. This was consistent with the observed analogue docked poses within the clathrin terminal domain Site 1 binding pocket. Further modifications targeted the naphthalene imide moiety, with the installation of 5-Br (45 a), 5-OH (45 c) and 5-propyl ether (45 d) moieties. Among them, the OH 45 c and propyl ether 45 d retained PPI inhibition, with propyl ether 45 d being the most active with a PPI inhibition IC=7.3 μM. This is 2x more potent than Pitstop 2 and 3x more potent than Pitstop 1.
合成了25种Pitstop 1和2的嵌合化合物，并对其阻断网格蛋白末端结构域-发动蛋白蛋白-蛋白相互作用（使用酶联免疫吸附测定法进行NTD-PPI）以及细胞中网格蛋白介导的内吞作用（CME）的能力进行了筛选。文库1基于Pitstop 2，但未观察到显著的网格蛋白PPI或细胞内活性。以Pitstop 1为基础，制备了16种以1,8-萘二甲酰亚胺为核心的类似物。具有亚甲基间隔连接基和简单酰胺的类似物显示出适度到良好的PPI抑制范围（分别为7.6-42.5 μM，萘基39和4-硝基苯基40）活性。这些数据揭示了萘磺酸盐部分的重要性，没有脱-SO类似物显示出PPI抑制作用。这与观察到的类似物在网格蛋白末端结构域位点1结合口袋内的对接姿势一致。进一步的修饰针对萘二甲酰亚胺部分，引入了5-Br（45 a）、5-OH（45 c）和5-丙基醚（45 d）部分。其中，OH 45 c和丙基醚45 d保留了PPI抑制作用，丙基醚45 d活性最高，PPI抑制IC=7.3 μM。这比Pitstop 2强2倍，比Pitstop 1强3倍。
