乳腺癌治疗一直是一个全球性难题，基于缺氧肿瘤微环境（TME）的靶向策略引起了广泛关注。作为缺氧肿瘤中过度表达的标志性转录因子，缺氧诱导因子 1 (HIF-1) 有助于癌症进展。化合物7-(3-(2-氯-1H-苯并[d]1咪唑-1-基)丙氧基)-2-(3,4,5-三甲氧基苯基)-4H-苯并吡喃-4-酮，合成并我们早期的研究将FB15命名为HIF-1α信号通路的潜在抑制剂，已被证明是多种癌症化疗的有前景的候选药物。然而，FB15的溶解度差和药代动力学不良导致其对肿瘤的治疗效果有限，最终限制了其潜在的临床应用。碳酸酐酶 IX (CAIX) 是一种肿瘤细胞跨膜蛋白，在缺氧肿瘤部位过度表达。乙酰唑酰胺 (AZA) 是一种靶向 CAIX 的高选择性配体，可用于将 FB15 递送至缺氧肿瘤部位。在本研究中，我们制备并表征了 AZA 缀合的泊洛沙姆 188 (AZA-P188) 和 D-a 负载 FB15 的纳米混合胶束。 -生育酚聚乙烯1000乙二醇琥珀酸酯(TPGS)，表示为AZA-P188/TPGS@FB15。通过体外药物释放、细胞毒性测定、细胞摄取以及体内药代动力学和荧光成像评估其体外和体内递送效率。最后，利用临床前乳腺癌皮下移植模型研究了AZA-P188/TPGS@FB15的体内治疗效果。体外研究表明，AZA-P188/TPGS@FB15可以有效靶向CAIX受体介导的乳腺癌细胞，触发FB15响应酸性条件释放，并增强细胞摄取和针对乳腺癌细胞的细胞毒性。药代动力学研究表明，负载 FB15 的 AZA 功能化胶束表现出比游离 FB15 显着增加的 AUC0-t。体内成像表明，AZA 功能化胶束显着增加了肿瘤部位的药物分布。体内实验证实AZA-P188/TPGS@FB15对裸鼠肿瘤生长具有优异的抑制作用，且具有良好的生物安全性。AZA-P188/TPGS@FB15有望成为乳腺癌的潜在有效治疗途径。其利用 AZA 作为载体的靶向递送系统显示出提高 FB15 在癌症治疗中疗效的潜力。© 2024 Liu 等人。

Breast cancer treatment has been a global puzzle, and targeted strategies based on the hypoxic tumor microenvironment (TME) have attracted extensive attention. As a signature transcription factor overexpressed in hypoxia tumor, hypoxia-inducible factor-1 (HIF-1) contribute to cancer progression. Compound 7-(3-(2-chloro-1H-benzo[d]1midazole-1-yl) propoxy)-2-(3,4,5-trime-thoxyphenyl)-4H-chromen-4-one, synthesized and named FB15 in our earlier research, a potential inhibitor of HIF-1α signaling pathway, has been proved a promising drug candidate for many kinds of cancer chemotherapy. However, the poor solubility and undesirable pharmacokinetics of FB15 leads to limited treatment efficacy of tumor, which ultimately restricts its potential clinical applications. Carbonic anhydrase IX (CAIX), a tumor cell transmembrane protein, was overexpressed in hypoxia tumor site. Acetazolamide (AZA), a highly selective ligand targeting CAIX, can be utilized to delivery FB15 to hypoxia tumor site.In this study, we prepared and characterized FB15 loaded nano-mixed micelles with the AZA conjugated poloxamer 188 (AZA-P188) and D-a-Tocopherol Polyethylene 1000 Glycol Succinate (TPGS), denoted as, AZA-P188/TPGS@FB15. Its delivery efficiency in vitro and in vivo was assessed by in vitro drug release, cytotoxicity assay, cellular uptake, and in vivo pharmacokinetics and fluorescence imaging. Finally, therapeutic effect of AZA-P188/TPGS@FB15 was investigated using a preclinical breast cancer subcutaneous graft model in vivo.In vitro studies revealed that AZA-P188/TPGS@FB15 could efficiently target breast cancer cells mediated by CAIX receptor, trigger FB15 release in response to acidic condition, and enhance cellular uptake and cytotoxicity against breast cancer cells. The pharmacokinetic studies showed that FB15-loaded AZA-functionalized micelles exhibited significantly increased AUC0-t over free FB15. In vivo imaging demonstrated that AZA-functionalized micelles significantly increased the drug distribution in the tumor site. In vivo experiments confirmed that AZA-P188/TPGS@FB15 exhibited superior inhibition of tumor growth in nude mice with good biosafety.AZA-P188/TPGS@FB15 hold promise as a potentially effective therapeutic way for breast cancer. Its targeted delivery system utilizing AZA as a carrier shows potential for improving the efficacy of FB15 in cancer therapy.© 2024 Liu et al.