白蛋白结合肽-多柔比星耦合物的设计、制备、表征及初步评价  

Design,Preparation,Characterization and Preliminary Evaluation of an Albumin Binding Peptide-Doxorubicin Conjugate

作　　者：刘立平[1] 张纯[2] 殷爽[2] 王祺[2] 张耀[2] 余蓉[1] 刘永东[2] 苏志国[2] 

机构地区：[1]四川大学华西药学院靶向药物及释药系统教育部重点实验室,成都610041 [2]中国科学院过程工程研究所生化工程国家重点实验室,北京100190

出　　处：《中国生物工程杂志》2017年第4期68-75,共8页China Biotechnology

基　　金：国家自然科学基金(21576267);教育部博士点基金(120120181110036);生化工程国家重点实验室开放基金(2014KF-05);北京市自然科学基金(2162041)资助项目

摘　　要：为了改善多柔比星在血液中的循环半衰期,基于白蛋白在血浆中含量丰富、性质稳定、半衰期持久的特点,通过大肠杆菌体系重组表达了源于细菌表面蛋白中白蛋白结合肽段,并建立了纯化工艺,获得了纯度高于95%的重组白蛋白结合多肽,并证明重组多肽在体外能够迅速地与人血清白蛋白结合。通过与DOXO-EMCH分子化学耦联,制备获得重组白蛋白结合肽-多柔比星耦合物。A549细胞毒力实验结果显示耦合物的IC50浓度升高,药代动力学结果显示重组白蛋白结合肽能够显著延长多柔比星在血液中的循环半衰期,相对于多柔比星或DOXO-EMCH分别提高了5.6倍和3.8倍。荷瘤小鼠肿瘤生长抑制实验结果显示耦合物的抗肿瘤效力得到提升。证明了基于白蛋白结合机制可为小分子药物长效设计提供新的思路。Doxorubicin is a widely used anthracycline that of many types of hematologic and solid cancers. The clinical limited by its severe side effects, including neutropenia, has been proven highly effective in the treatment application of this antitumor drug is, however, mucositis, myelosuppression and cumulative cardiotoxicity. Besides, the therapeutic potential of this agent is also significantly limited due to the defects of short circulation time and quick clearance. In order to improve the therapeutic efficacy of doxorubicin, there is a need for developing effective strategies to prolong its blood circulation time. Currently, a novel long-term approach based on albumin is widely studied. Albumin is the most abundant and stable protein in plasma, which can maintain an extraordinarily long circulatory half-life of 15 ~ 19 days due to its size which is bigger than the renal filtration threshold and its interaction with the FcRn-mediated recycling. Based on these characteristics, albumin is thus regarded as a perfect vehicle to be used to extend the circulatory half-life of drugs which can be conjugated or genetically fused to albumin. Besides, proteins that specifically bind to albumin have also been employed to extend the half-life of therapeutic agents. Albumin-binding domain, a novel short peptide derived from bacterial surface proteins, is a naturally occurring left-handed three-helix bundle which has an extremely high affinity for HSA in the femtomolar range. It is reported that the helices 2 and 3 in surface exposed side chains of this peptide are able to selectively bind to the domain II of serum albumin. The pharmacokinetic properties of many sorts of drugs can be easily improved by conjugating to this albumin-binding module. In order to improve the circulating half-life of doxorubicin in blood, a new kind of cysteine-containing albumin-binding domain was firstly expressed in E. coli and successfully prepared through a two-step chromatography with a purity of above 95%. And it is confirmed that this recombina

关 键 词：多柔比星 白蛋白结合肽 血清白蛋白 长效性 抗肿瘤 

分 类 号：Q819[生物学—生物工程]