Aptamer Functionalized Hypoxia-potentiating Agent and Hypoxia-inducible Factor Inhibitor Combined with Hypoxia-activated Prodrug for Enhanced Tumor Therapy

Triple-negative breast cancer (TNBC) is the most lethal subtype of breast cancer. Hypoxiaactivated prodrugs (HAPs) have shown promise as potential therapeutic agents for TNBC. While increasing hypoxia levels may promote the HAP activation, it raises concerns regarding HIF1α-dependent drug resistance. It is desirable to develop a targeted approach that enhances tumor hypoxia for HAP activation without promoting HIF1α-dependent drug resistance in TNBC treatment. Herein, we proposed a multi-responsive carrier-free selfassembled nanomedicine named AQ4N@CA4T1ASO. This nanomedicine first targeted tumors by the TNBC-targeting aptamers (T1), and then disassembled in the reductive and acidic conditions within tumors. The released Combretastatin 4 (CA4) could exacerbate hypoxia, thereby promoting the conversion of inactive Banoxantrone (AQ4N) to its active form, AQ4. Simultaneously, the released antisense oligonucleotide (ASO) could attenuate hypoxia-induced HIF1α mRNA expression, thereby sensitizing the tumor to chemotherapy. Overall, this smart nanomedicine represents a profound targeted therapy strategy, combining "hypoxia-potentiating, hypoxia-activated, chemo-sensitization" approaches for TNBC treatment. In vivo study demonstrated significant suppression of tumor growth, Revised Manuscript with track changes Click here to view linked References 2 highlighting the promising potential of this nanomedicine for future clinical translation.

三阴性乳腺癌（Triple-negative breast cancer, TNBC）是致死性最强的乳腺癌亚型。缺氧激活前药（Hypoxia-activated prodrugs, HAPs）作为TNBC的潜在治疗制剂已展现出应用前景。尽管升高的肿瘤缺氧水平可促进HAPs活化，但同时也会引发依赖于缺氧诱导因子1α（HIF1α）的耐药性问题，因此亟需开发一种靶向策略，在TNBC治疗中既能增强肿瘤缺氧以实现HAPs活化，又不会诱导依赖HIF1α的耐药性。在此，我们提出了一种多响应无载体自组装纳米药物（multi-responsive carrier-free selfassembled nanomedicine），命名为AQ4N@CA4T1ASO。该纳米药物首先通过三阴性乳腺癌靶向适配体（T1）靶向肿瘤，随后在肿瘤内部的还原及酸性微环境中发生解离。释放出的康普瑞汀4（Combretastatin 4, CA4）可加剧肿瘤缺氧，进而将非活性形式的班诺腙（Banoxantrone, AQ4N）转化为活性形式AQ4。与此同时，释放出的反义寡核苷酸（antisense oligonucleotide, ASO）可抑制缺氧诱导的HIF1α mRNA表达，从而使肿瘤对化疗更加敏感。总体而言，这种智能纳米药物代表了一种极具潜力的靶向治疗策略，整合了“缺氧强化、缺氧激活、化疗增敏”三种手段用于TNBC治疗。体内（in vivo）研究证实其可显著抑制肿瘤生长；带修订轨迹的修订稿点击此处查看关联参考文献2，凸显了该纳米药物在未来临床转化中的可观应用潜力。