新型靶向HER3抗体偶联药物的抗肿瘤活性研究  

Antitumor activity of a novel HER3-targeting antibody-drug-conjugate

作　　者：耿晶 李新颖 GENG Jing;LI Xin-ying(Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China;State Key Laboratory of Toxicology and Medical Countermeasures,Beijing Institute of Pharmacology and Toxicology,Beijing 100850,China)

机构地区：[1]中国医学科学院医药生物技术研究所,北京100050 [2]军事医学研究院毒物药物研究所,毒理学与医学对策国家重点实验室,北京100850

出　　处：《药学学报》2022年第6期1825-1831,共7页Acta Pharmaceutica Sinica

基　　金：中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-026)。

摘　　要：作为人表皮生长因子受体(epidermal growth factor receptor,EGFR,HER1,ERBB)家族重要成员,人表皮生长因子受体3(HER3,ERBB3)是PI3K/AKT信号通路中关键的激活因子。近期研究发现,HER3与多种类型肿瘤的发生发展以及对EGFR和人表皮生长因子受体2(HER2,ERBB2)疗法的获得性耐药密切相关;但利用靶向HER3的中和抗体治疗肿瘤的效果并不理想,这很可能是由于靶向HER3的中和抗体并不能完全阻断异源二聚体形成。抗体-化学药物偶联物(antibody-drug conjugates,ADCs)可通过抗体特异性结合靶细胞,从而定向发挥高细胞毒性化学药物对癌细胞高度杀伤的效果,已被广泛应用于临床治疗中。本研究通过计算机辅助分子模拟技术对HER3与前期获得的抗体LmAb3的复合物结构进行模拟优化,利用距离几何学、计算机图形学技术预测了LmAb3中参与抗原结合的关键位点,利用点突变技术获得了新型抗HER3高亲和力抗体FD001。亲和力测定结果显示,FD001、LmAb3与HER3结合的解离常数KD值分别为1.48E-11与2.46E-10,表明FD001具有更强的靶向结合HER3的能力。通过赖氨酸偶联技术将FD001与美登素(DM1)偶联获得抗体药物偶联物FD001-DM1。细胞实验结果显示,FD001-DM1能够有效抑制HER3阳性HT-29结肠癌细胞增殖,半数有效浓度(median effective concentration,EC_(50))为32.23 nmol·L^(-1)。利用建立的小鼠肿瘤模型进行抗肿瘤活性评价,实验结果显示FD001-DM1治疗组肿瘤体积约为对照组的25%,且组内小鼠体重无明显下降,FD001-DM1具有良好的体内外抗肿瘤活性及安全性。本研究为探寻基于HER3的新型ADCs药物提供了有效的帮助。本研究中的小鼠按照国际实验室动物护理和使用准则进行使用和治疗,并得到了军事医学研究院军事认知与脑科学研究所动物伦理委员会的批准。As a member of the human epidermal growth factor receptor(HER)family of receptor tyrosine kinases,HER3 is an aberrantly activator of the PI3K/AKT pathway.Studies have indicated that HER3 is related to the progression of a variety of tumor types such as breast cancer,non-small cell lung cancer(NSCLC),ovary cancer and colon cancer,and in acquired resistance to EGFR and HER2 therapies.However,the attempts to target HER3 with neutralizing antibodies are not ideal previously.This is most likely due to the fact that the antibodies targeting HER3 fail to completely block the heterodimerization of HER3 and other receptors.Antibody-drug conjugates(ADCs)can specifically bind to target cells and exert the highly cytotoxicity effect on cancer cells through chemical drugs.ADCs have been widely used in clinical cancer therapies.We analyzed and optimized the structure of the antigen-antibody complex between HER3 and antibody LmAb3 by computer-aided molecular simulation technology,and the key sites involved in antigen binding in LmAb3 were predicted by distance geometry and computer graphics technology.Then a novel anti-HER3 antibody FD001 was obtained by point mutation technology.The affinity measurement by ForteBio results showed that the affinity of FD001 is much higher than LmAb3,the K_(D) values of FD001 and LmAb3 with HER3 were 1.48E-11 and 2.46E-10,respectively.Antibody drug conjugate FD001-DM1 is obtained by coupling FD001 to DM1[emtansine,N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-maytansine]by lysine coupling technology.The results of cell cytotoxicity experiments showed that FD001-DM1 could effectively inhibit the proliferation of HER3-positive HT-29 colon cancer cells,with EC_(50) value of 33.62 nmol·L^(-1).The in vivo xenografts therapy results showed that the tumor volume of the FD001-DM1 treatment group was about 25%of that of the control group,and there was no significant weight reduction of the mice.These results reveal that FD001-DM1 had good in vivo and in vitro anti-tumor activity with high safety,which may p

关 键 词：人表皮生长因子受体家族 HER3 单克隆抗体 抗体-药物偶联物 计算机辅助分子模建 

分 类 号：R966[医药卫生—药理学]