天然产物（NP）在治疗药物的开发中发挥着至关重要的作用。然而，确定纳米粒子的靶标仍然具有很大的挑战性。此外，纳米粒子通常通过作用于多个靶点或途径来发挥药理活性，这也给纳米粒子的靶点识别带来了很大的困难。受我们在设计类药物蛋白降解剂方面不断努力的启发，本研究介绍了该靶点的成功例子利用PROTAC技术对天然产物吴茱萸碱进行鉴定和药物发现。利用蛋白水解靶向嵌合体（PROTAC）的优势，本文开发了一种结合PROTAC衍生化、定量蛋白质组分析和结合亲和力验证的综合策略，用于抗肿瘤NP吴茱萸碱的靶标鉴定和药物发现在这项研究中，高效的 PROTAC 和阴性对照均被设计用于定量蛋白质组分析。此外，REXO4 被证实是 3-氟-10-羟基左旋二胺的直接靶标，可通过 ROS 诱导细胞死亡。此外，PROTAC 13c在体外和体内均能有效降解REXO4，从而产生有效的抗肿瘤活性并减少毒副作用。综上所述，我们开发了纳米颗粒的靶点识别和药物发现的综合策略，并成功应用于吴茱萸碱的PROTAC衍生化和靶点表征。这项概念验证研究凸显了 PROTAC 技术在 NP 靶标识别方面的优越性，加速了基于 NP 的药物发现进程，展示了在基于 NP 的药物开发中的广泛应用。版权所有 © 2023。由 Elsevier B.V. 制作和托管。

Natural products (NPs) play a crucial role in the development of therapeutic drugs. However, it is still highly challenging to identify the targets of NPs. Besides, NPs usually exert their pharmacological activities via acting on multiple targets or pathways, which also poses great difficulties for the target identification of NPs.Inspired by our continuous efforts in designing drug-like protein degraders, this study introduced a successful example for the target identification and drug discovery of natural products evodiamine by employing PROTAC technology.Taking advantages of proteolysis targeting chimera (PROTAC), herein an integrated strategy combining PROTAC derivatization, quantitative proteomic analysis and binding affinity validation was developed for target identification and drug discovery of antitumor NP evodiamine.In this study, both highly potent PROTACs and negative controls were designed for quantitative proteomic analysis. Furthermore, REXO4 was confirmed as a direct target of 3-fluoro-10-hydroxylevodiamine, which induced cell death through ROS. In addition, the PROTAC 13c effectively degraded REXO4 both in vitro and in vivo, leading to potent antitumor activity and reduced toxic side effects.In summary, we developed an integrated strategy for the target identification and drug discovery of NPs, which was successfully applied to the PROTAC derivatization and target characterization of evodiamine. This proof-of-concept study highlighted the superiority of PROTAC technology in target identification of NPs and accelerated the process of NPs-based drug discovery, exhibiting broad application in NP-based drug development.Copyright © 2023. Production and hosting by Elsevier B.V.