由于其肿瘤异质性，三阴性乳腺癌（TNBC）的治疗至今仍是一个巨大的挑战。目前，临床上仍缺少有效的针对TNBC的靶向治疗手段。顺铂（Cisplatin， Cis）作为常用的广谱化疗药物，在TNBC中显示出良好的治疗效果。为对抗肿瘤的异质性，大量的研究尝试通过Cis与各种激酶抑制剂的联合治疗，以多重抑瘤机制来治疗TNBC。其中，Cis与SRC抑制剂达沙替尼（Dasatinib，DAS）的联合使用初步显示出了良好的抗TNBC疗效，但与此同时也观察到较为严重的毒副作用。为了克服这种双药物联合治疗带来的严重毒性，我们通过自组装策略将Cis和DAS共负载到透明质酸（Hyaluronan，HA）纳米凝胶中（HA/Cis/DAS，命名为HCD纳米凝胶），用于实现TNBC靶向的Cis与DAS共递送与治疗应用。研究发现，所获得的HCD纳米凝胶在水溶液中的水合粒径约为181 nm，其形貌均一，可以长期在水溶液环境中储存，同时在含10 %胎牛血清的PBS溶液环境中也可以保持12 h以上的体系稳定。在细胞水平上，HCD纳米凝胶显示出了Cis和DAS的双重药理活性。此外，在谷胱甘肽（GSH）水平升高（10 mM）的情况下，HCD纳米凝胶表现出快速的药物释放和颗粒聚集，证明HCD纳米凝胶在高GSH的肿瘤微环境中具有响应性药物释放潜力。我们还通过竞争性实验证实了HCD纳米凝胶可以利用HA对TNBC细胞上过表达的CD44的靶向能力，实现针对TNBC细胞的靶向药物递送。在动物实验中，我们使用4T1细胞构建了TNBC皮下瘤模型小鼠，验证了HCD纳米凝胶的靶向共递送与减毒能力。在保持Cis和DAS联合用药的良好抗癌效果前提下，HCD纳米凝胶显著提高了组合药物的最大耐受剂量，延长了药物的体内循环时间。通过肾脏组织切片和免疫荧光染色实验，我们观察到HCD纳米凝胶组的肾毒性大幅降低。

综上所述，本研究制备的HCD纳米凝胶在4T1肿瘤模型中，显著改善了Cis和DAS联合治疗方案，为TNBC的靶向治疗提供了一种新的潜在治疗策略。

Due to the tumor heterogeneity, the treatment of triple-negative breast cancer (TNBC) is still a great challenge, and there is currently no effective targeted therapy for TNBC in clinical practice. Cisplatin (Cis), as a commonly used broad-spectrum chemotherapeutic drug, has shown a good therapeutic effect in TNBC. Some studies have tried to combat the heterogeneity of TNBC by combining Cis with various kinase inhibitors, among which the combination of Cis and SRC inhibitor dasatinib (DAS) has initially shown good anti-TNBC efficacy, but at the same time, serious toxic side effects have also been observed. In order to overcome the serious toxicity caused by this dual chemotherapeutic drug combination, Cis and DAS were co-loaded into hyaluronic acid (HA) nanogel (HA/Cis/DAS, named as HCD nanogel) by a self-assembly strategy. The obtained HCD nanogel had a hydrodyanmic size of about 181 nm in aqueous solution and a uniform morphology. The HCD can be stored in aqueous solution for a long time, and can also be kept stable for more than 12 hours in PBS solution containing 10 % fetal bovine serum. HCD nanogels exhibited dual pharmacological activities of both Cis and DAS at the cellular level. In addition, HCD nanogels exhibited accelerated drug release and particle aggregation in the presence of elevated glutathione (GSH) levels (10 mM), demonstrating the responsive drug release potential of HCD nanogels in the tumor microenvironment with high GSH. Based on the competition assay with the presence of extra free HA, we also confirmed that HCD nanogels can utilize the ability of HA to target CD44 overexpressed on TNBC cells to achieve targeted drug delivery against TNBC cells. In animal experiments, we used 4T1 cells to construct a TNBC mouse model, and verified the targeted co-delivery ability as well as alleviated side effects of HCD nanogels. Other than maintaining the effective anticancer effect of Cis and DAS combination, the maximum tolerated dose of dual chemotherapy drugs was significantly increased, and the circulation time of drugs in vivo was prolonged by utilizing the HCD nanogels. By immunofluorescence staining of kidney tissue sections, we observed a significant reduction of nephrotoxicity in the HCD nanogel group.

In conclusion, the HCD nanogel prepared in this study improves the Cis and DAS combination treatment regimen and potentially provides a new targeted therapy strategy for the targeted treatment of TNBC.