Selvita Ltd., Zagreb, Croatia.
Ruđer Bošković Institute, Zagreb, Croatia.
J Enzyme Inhib Med Chem. 2023 Dec;38(1):2183809. doi: 10.1080/14756366.2023.2183809.
The process of identifying the protein targets and off-targets of a biologically active compound is of great importance in modern drug discovery. Various chemical proteomics approaches have been established for this purpose. To compare the different approaches, and to understand which method would provide the best results, we have chosen the EGFR inhibitor lapatinib as an example molecule. Lapatinib derivatives were designed using linkers with motifs, including amino (amidation), alkyne (click chemistry) and the diazirine group (photo-affinity). These modified lapatinib analogues were validated for their ability to inhibit EGFR activity and were shown to pull down purified recombinant EGFR protein. In all of the approaches evaluated here, we identified EGFR as the main protein target from the lysate of immortalised cell line expressing EGFR, thus validating its potential use to identify unknown protein targets. Taken together, the results reported here give insight into the cellular activities of lapatinib.
确定生物活性化合物的蛋白质靶标和脱靶的过程在现代药物发现中非常重要。为此，已经建立了各种化学蛋白质组学方法。为了比较不同的方法，并了解哪种方法会提供最佳结果，我们选择 EGFR 抑制剂拉帕替尼作为示例分子。使用带有基序的接头设计了拉帕替尼衍生物，包括氨基（酰胺化）、炔基（点击化学）和重氮化物基团（光亲和）。这些修饰的拉帕替尼类似物经过验证，能够抑制 EGFR 的活性，并能够拉下纯化的重组 EGFR 蛋白。在评估的所有方法中，我们从表达 EGFR 的永生化细胞系的裂解物中鉴定出 EGFR 作为主要蛋白质靶标，从而验证了其用于鉴定未知蛋白质靶标的潜力。总之，这里报道的结果深入了解了拉帕替尼的细胞活性。
