Neijssen Joost, Cardoso Rosa M F, Chevalier Kristen M, Wiegman Luus, Valerius Thomas, Anderson G Mark, Moores Sheri L, Schuurman Janine, Parren Paul W H I, Strohl William R, Chiu Mark L
Genmab, Utrecht, The Netherlands.
Janssen Research & Development, Spring House, Pennsylvania, USA.
J Biol Chem. 2021 Jan-Jun;296:100641. doi: 10.1016/j.jbc.2021.100641. Epub 2021 Apr 8.
A bispecific antibody (BsAb) targeting the epidermal growth factor receptor (EGFR) and mesenchymal-epithelial transition factor (MET) pathways represents a novel approach to overcome resistance to targeted therapies in patients with non-small cell lung cancer. In this study, we sequentially screened a panel of BsAbs in a combinatorial approach to select the optimal bispecific molecule. The BsAbs were derived from different EGFR and MET parental monoclonal antibodies. Initially, molecules were screened for EGFR and MET binding on tumor cell lines and lack of agonistic activity toward MET. Hits were identified and further screened based on their potential to induce untoward cell proliferation and cross-phosphorylation of EGFR by MET via receptor colocalization in the absence of ligand. After the final step, we selected the EGFR and MET arms for the lead BsAb and added low fucose Fc engineering to generate amivantamab (JNJ-61186372). The crystal structure of the anti-MET Fab of amivantamab bound to MET was solved, and the interaction between the two molecules in atomic details was elucidated. Amivantamab antagonized the hepatocyte growth factor (HGF)-induced signaling by binding to MET Sema domain and thereby blocking HGF β-chain-Sema engagement. The amivantamab EGFR epitope was mapped to EGFR domain III and residues K443, K465, I467, and S468. Furthermore, amivantamab showed superior antitumor activity over small molecule EGFR and MET inhibitors in the HCC827-HGF in vivo model. Based on its unique mode of action, amivantamab may provide benefit to patients with malignancies associated with aberrant EGFR and MET signaling.
一种靶向表皮生长因子受体（EGFR）和间充质上皮转化因子（MET）通路的双特异性抗体（BsAb）代表了一种克服非小细胞肺癌患者对靶向治疗耐药性的新方法。在本研究中，我们采用组合方法对一系列BsAb进行了序贯筛选，以选择最佳的双特异性分子。这些BsAb源自不同的EGFR和MET亲本单克隆抗体。最初，在肿瘤细胞系上筛选分子与EGFR和MET的结合情况，以及对MET缺乏激动活性。鉴定出命中分子，并基于它们在无配体情况下通过受体共定位诱导MET对EGFR的不良细胞增殖和交叉磷酸化的可能性进行进一步筛选。在最后一步之后，我们为先导BsAb选择了EGFR和MET臂，并添加了低岩藻糖Fc工程以生成阿米万他单抗（JNJ-61186372）。解析了与MET结合的阿米万他单抗抗MET Fab的晶体结构，并阐明了两个分子之间原子细节上的相互作用。阿米万他单抗通过结合MET Sema结构域拮抗肝细胞生长因子（HGF）诱导的信号传导，从而阻断HGFβ链-Sema结合。阿米万他单抗的EGFR表位被定位到EGFR结构域III以及残基K443、K465、I467和S468。此外，在体内HCC827-HGF模型中，阿米万他单抗显示出比小分子EGFR和MET抑制剂更强的抗肿瘤活性。基于其独特的作用模式，阿米万他单抗可能为与异常EGFR和MET信号传导相关的恶性肿瘤患者带来益处。
