具有更好生物分子递送特性的细胞穿透肽（CPP）将扩大其临床应用。利用MLCPP2.0机器算法，我们从核定位信号/核输出信号数据库中筛选出多个具有潜在细胞摄取能力的候选序列，并通过细胞穿透荧光示踪实验进行验证。源自山羊关节炎脑炎病毒 Rev 蛋白的肽 (NCR) 表现出高效的细胞穿透活性，比经典的 CPP TAT 传递的 EGFP 多四倍以上，使其能够在溶酶体中积累。结构和性质分析表明，高疏水矩和适当的疏水区域有助于 NCR 的高递送活性。 Trastuzumab emtansine (T-DM1) 是一种靶向 HER2 的抗体药物偶联物，可以通过提高靶向递送效率和增加溶酶体药物递送来提高其抗肿瘤活性。本研究设计了一种新的NCR载体，通过融合域Z非共价结合T-DM1，可以特异性结合免疫球蛋白G的Fc区，有效地将T-DM1递送至溶酶体。 MTT结果显示，域Z-NCR载体显着增强了T-DM1对HER2阳性肿瘤细胞的细胞毒性，同时保持了药物特异性。我们的结果为探索 CPP 作为溶酶体靶向递送工具的潜在应用做出了有益的尝试。© 2024 欧洲肽协会和 John Wiley

Cell-penetrating peptides (CPPs) with better biomolecule delivery properties will expand their clinical applications. Using the MLCPP2.0 machine algorithm, we screened multiple candidate sequences with potential cellular uptake ability from the nuclear localization signal/nuclear export signal database and verified them through cell-penetrating fluorescent tracing experiments. A peptide (NCR) derived from the Rev protein of the caprine arthritis-encephalitis virus exhibited efficient cell-penetrating activity, delivering over four times more EGFP than the classical CPP TAT, allowing it to accumulate in lysosomes. Structural and property analysis revealed that a high hydrophobic moment and an appropriate hydrophobic region contribute to the high delivery activity of NCR. Trastuzumab emtansine (T-DM1), a HER2-targeted antibody-drug conjugate, could improve its anti-tumor activity by enhancing targeted delivery efficiency and increasing lysosomal drug delivery. This study designed a new NCR vector to non-covalently bind T-DM1 by fusing domain Z, which can specifically bind to the Fc region of immunoglobulin G and effectively deliver T-DM1 to lysosomes. MTT results showed that the domain Z-NCR vector significantly enhanced the cytotoxicity of T-DM1 against HER2-positive tumor cells while maintaining drug specificity. Our results make a useful attempt to explore the potential application of CPP as a lysosome-targeted delivery tool.© 2024 European Peptide Society and John Wiley & Sons Ltd.