本研究采用药物重新定位策略来发现新型 PD-L1 小分子抑制剂。建立了 3D-QSAR 药效团模型，随后通过各种手段进行验证，以选择适合虚拟筛选的稳健模型 Hypo-1。 Hypo1 用于筛选 Drugbank 数据库中的 7,475 种化合物，与 PD-L1 分子对接后鉴定出 283 种分子。19 种化合物进行了 HTRF 检测，其中 15 种显示出不同程度的 PD-1/PD- 抑制作用。 L1 交互。使用SPR实验进一步证实化合物2202、2204、2207和2208与PD-L1结合。其中，化合物2204（Daclatasvir，KD=11.4μM）对人PD-L1的亲和力比对照化合物BMS-1更高。在HepG2/Jurkat细胞共培养模型中，Daclatasvir有效激活Jurkat细胞以杀死HepG2细胞。在小鼠 H22 肝细胞肿瘤模型中，达卡他韦显着抑制肿瘤生长（剂量为 100 mg/kg 时，TGI = 53.4%）。与仑伐替尼联合使用时，其抗肿瘤作用更加明显（TGI = 85.1%）。脾细胞和肿瘤细胞的流式细胞术分析表明达卡他韦在两种模型中均激活了免疫系统。总之，Daclatasvir 被确定为一种新型 PD-L1 小分子抑制剂。版权所有 © 2024 Elsevier Inc. 保留所有权利。

This study employed a drug repositioning strategy to discover novel PD-L1 small molecule inhibitors. 3D-QSAR pharmacophore models were establishedand subsequently validated through various means to select a robust model, Hypo-1, suitable for virtual screening. Hypo1 was used toscreen a library of 7,475 compounds from the Drugbank database, leading to the identification of 283 molecules following molecular docking with PD-L1.19 compounds underwent HTRF assays, with 15 showing varying degrees of inhibition of the PD-1/PD-L1 interaction. Compounds2202,2204,2207, and2208were further confirmed to bind to PD-L1 using SPR experiments. Among them, compound2204(Daclatasvir, KD = 11.4 μM) showeda higher affinity for human PD-L1 than the control compound BMS-1. In the HepG2/Jurkat cell co-culture model, Daclatasvir effectively activated Jurkat cells to kill HepG2 cells. In the mouse H22 hepatocellular tumor model, Daclatasvir significantly inhibited tumor growth (TGI = 53.4 % at a dose of 100 mg/kg). Its anti-tumor effect was more pronounced when combined with Lenvatinib (TGI = 85.1 %). Flow cytometry analysis of splenocytes and tumor cells indicated that Daclatasvir activated the immune system in both models. In summary, Daclatasvir was identified as a novel PD-L1small molecule inhibitor.Copyright © 2024 Elsevier Inc. All rights reserved.