Student Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran; Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
Int J Pharm. 2022 Jul 25;623:121892. doi: 10.1016/j.ijpharm.2022.121892. Epub 2022 Jun 4.
Doxorubicin (DOX) is a chemotherapeutic drug that belongs to the anthracyclines family. Cardiotoxicity is one of the main limiting factor of prescribing DOX. To reduce its side effects and enhance the drug delivery to the targeted tissues, we aimed to establish a new targeted and controlled release drug delivery system for treatment of breast cancer. In this article, we tried to synthesize a new nanoplatform consisted of DOX conjugate with hydrazide and disulfide bonds to the hyaluronic acid (HA). Firstly, 4,4'-Dithiodibutyric acid (DTBH) was conjugated with HA. Then, 3-aminophenyl boronic acid monohydrate (APBA) was conjugated with DTBH-HA. Subsequently, DOX was added to DTBH-HA-APBA. HA is a natural polymer with the ability to target CD44, a cell surface adhesion receptor, which are highly overexpressed on the surface of variety of cancer cells. Other targeting agent, APBA can target sialic acid on the cancer cells surface and improve the tumor uptake. Formation of The DTBH-HA-APBA conjugate was confirmed by proton nuclear magnetic resonance (H NMR) spectroscopy. Scanning emission electron microscopy (SEM) images of the DOX-DTBH-HA-APBA displayed a spherical shape with an average diameter of about 70 nm. In vitro drug release study showed considerably different release pattern of DOX from the formulation at acidic pH (5.4) which was higher than normal pH (7.4). Cellular uptake and cellular cytotoxicity analysis were examined in human breast adenocarcinoma cell line (MCF-7) and mouse breast cancer cells (4T1) as positive cell lines and Chinese Hamster Ovary cells (CHO) as negative cell line. Results confirmed that there is a remarkable difference between dual-targeted (DOX-DTBH-HA-APBA) and single targeted (DOX-DTBH-HA) formulations in both positive cell lines regarding internalization and cytotoxicity. In vivo studies indicated that dual-targeted formulation has the best efficacy with minimum side effects in mouse model. Fluorescence imaging of organs revealed that DOX-DTBH-HA-APBA showed greater DOX accumulation compared with DOX-DTBH-HA and free DOX in tumor site. Also, pathological evaluation indicated that there is no observable cardiotoxicity with final formulation.
多柔比星（DOX）是一种属于蒽环类的化疗药物。心脏毒性是开具 DOX 的主要限制因素之一。为了降低其副作用并增强药物向靶向组织的输送，我们旨在建立一种新的针对乳腺癌的靶向和控制释放药物输送系统。在本文中，我们尝试合成一种由 DOX 与酰肼和二硫键缀合的新型纳米平台到透明质酸（HA）。首先，将 4,4'-二硫代二丁酸（DTBH）与 HA 缀合。然后，将 3-氨基苯硼酸一水合物（APBA）与 DTBH-HA 缀合。随后，将 DOX 添加到 DTBH-HA-APBA 中。HA 是一种具有靶向 CD44 能力的天然聚合物，CD44 是一种细胞表面粘附受体，在多种癌细胞表面高度过表达。另一种靶向剂 APBA 可以靶向癌细胞表面的唾液酸并提高肿瘤摄取率。通过质子核磁共振（H NMR）光谱证实了 DTBH-HA-APBA 缀合物的形成。DOX-DTBH-HA-APBA 的扫描发射电子显微镜（SEM）图像显示出平均直径约为 70nm 的球形。体外药物释放研究表明，在酸性 pH（5.4）下，DOX 的释放模式与制剂有很大不同，高于正常 pH（7.4）。在人乳腺癌腺癌细胞系（MCF-7）和小鼠乳腺癌细胞（4T1）作为阳性细胞系和中国仓鼠卵巢细胞（CHO）作为阴性细胞系中进行了细胞摄取和细胞毒性分析。结果证实，在阳性细胞系中，双靶向（DOX-DTBH-HA-APBA）和单靶向（DOX-DTBH-HA）制剂在摄取和细胞毒性方面存在显著差异。体内研究表明，双靶向制剂在小鼠模型中具有最佳疗效且副作用最小。器官荧光成像显示，与 DOX-DTBH-HA 和游离 DOX 相比，DOX-DTBH-HA-APBA 在肿瘤部位具有更高的 DOX 积累。此外，病理评估表明，最终制剂没有可观察到的心脏毒性。
