摘要
利用定点原位生长技术,合成了一种温度响应性的干扰素-聚(2-甲基-2-丙烯酸-2-(2-甲氧基乙氧基)乙酯)偶联物(IFN-PDEGMA).当温度低于22oC时,IFN-PDEGMA处于溶解状态;当温度高于22oC时,IFN-PDEGMA则会发生聚集.由于小鼠体温高于22oC,因此,将IFN-PDEGMA原位注射到小鼠肿瘤组织处之后,它会在肿瘤组织处聚集驻留.体外实验结果显示,IFN-PDEGMA有效保持了干扰素的结构和活性;动物实验结果显示,相比于原药IFN-α,非温敏性IFN-POEGMA、商业化的长效干扰素PEGASYS以及IFN-PDEGMA可以更好地聚集驻留在肿瘤处,荷黑色素瘤小鼠的生存时间得以显著延长,分子量为10 kD a、30 kD a、60 k Da、100 kD a的IFN-PDEGMA所治疗的小鼠,其生存时间分别为36.5、31、29.5、28天,其中,10 kD a和30 kD a的IFN-PDEGMA的治疗效果均要优于PEGASYS.同时,生物安全性实验显示IFN-PDEGMA对正常组织器官不存在显著的毒副作用.
Interferon-α（IFN） has a short circulating half-life, which not only leads to limited clinical efficacy, but also causes severe side effects and poor patient compliance. Previously, we developed ELPfusion and site-specific in situ growth（SIG） methods to prolong the half-life of IFN, while the adopted intravenous administration still could not well concentrate IFN inside tumour tissues. In order to enhance the tumour accumulation and antitumour efficacy of IFN, in this study, we report intratumoural administration of temperature responsive interferon-poly（di（ethylene glycol） methyl ether methacrylate） conjugates（IFN-PDEGMA）. First, we employed SIG method to synthesize a series of temperature responsive IFN-PDEGMAs with different molecular weights（10 k Da, 30 k Da, 60 k Da and 100 k Da）. The preparation pr℃ess was monitored by SDS-PAGE, and the molecular weights, hydrodynamic radius and secondary structures of conjugates were characterized by GPC, DLS and CD（circular dichroism）, respectively. The phase transition temperatures were determined to be around 22 ℃. Thus, IFN-PDEGMA were soluble below 22 ℃; and they became insoluble above 22 ℃. Since the body temperature of mice is above 22 ℃, IFN-PDEGMA injected into the tumour tissue of mice aggregated l℃ally and became a drug depot in the tumour. In vitro characterization demonstrated that the structure and anti-proliferative activity of IFN were well remained for IFN-PDEGMA. In vivo experiments showed that the survival time of A375 melanoma-bearing mice were well extended by IFN-PDEGMA treatment compared with IFN-α. To be specific, the survival times of mice treated by IFN-PDEGMA of 10 k Da, 30 k Da, 60 k Da and 100 k Da were 36.5, 31, 29.5 and 28 days, respectively. IFN-PDEGMA of 10 k Da and 30 k Da both exhibited better anti-melanoma efficacy than commercial long-acting interferon, PEGASYS. Meanwhile, the biological safety experiments also showed that IFN-PDEGMA treatment did not have obvious side effects on normal tissues. In summary, we, for the first time, reported intratumoural administration of temperature responsive IFN-PDEGMA and studied the rule about how the molecular weight impacts on their properties in vitro and in vivo. This study not merely displayed an instance of temperature responsive protein-polymer conjugates and their anti-tumour efficacy, but also provided inspiration to further build a series of smart protein-polymer conjugates and seek for their potential applications in the diagnosis and treatment of major diseases such as cancer, virosis, diabetes and cardiovascular disease.
出处
《高分子学报》
SCIE
CAS
CSCD
北大核心
2018年第1期90-98,共9页
Acta Polymerica Sinica
基金
国家自然科学基金(基金号21534006)资助项目
关键词
干扰素
蛋白质-高分子偶联物
原子转移自由基聚合
药物递送
温度响应
Interferon, Protein-polymer conjugate, Atom transfer radical polymerization, Drug delivery, Temperature responsiveness
作者简介
通讯联系人:高卫平,E-mail:gaoweiping@tsinghua.edu.cn
