Institute of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Avenue, Boston, MA, 02115, USA.
Nat Commun. 2022 Jul 30;13(1):4431. doi: 10.1038/s41467-022-32159-6.
Acquired resistance to cetuximab in colorectal cancers is partially mediated by the acquisition of mutations located in the cetuximab epitope in the epidermal growth factor receptor (EGFR) ectodomain and hinders the clinical application of cetuximab. We develop a structure-guided and phage-assisted evolution approach for cetuximab evolution to reverse EGFR- or EGFR-driven resistance without altering the binding epitope or undermining antibody efficacy. Two evolved cetuximab variants, Ctx-VY and Ctx-Y104D, exhibit a restored binding ability with EGFR, which harbors the most common resistance substitution, S492R. Ctx-W52D exhibits restored binding with EGFR harboring another common cetuximab resistance substitution, G465R (EGFR). All the evolved cetuximab variants effectively inhibit EGFR activation and downstream signaling and induce the internalization and degradation of EGFR and EGFR as well as EGFR. The evolved cetuximab variants (Ctx-VY, Ctx-Y104D and Ctx-W52D) with one or two amino acid substitutions in the complementarity-determining region inherit the optimized physical and chemical properties of cetuximab to a great extent, thus ensuring their druggability. Our data collectively show that structure-guided and phage-assisted evolution is an efficient and general approach for reversing receptor mutation-mediated resistance to therapeutic antibody drugs.
获得性结直肠癌对西妥昔单抗的耐药性部分是由表皮生长因子受体 (EGFR) 胞外结构域中位于西妥昔单抗表位的突变获得介导的，这阻碍了西妥昔单抗的临床应用。我们开发了一种基于结构的和噬菌体辅助进化方法来进化西妥昔单抗，以逆转 EGFR 或 EGFR 驱动的耐药性，而不改变结合表位或破坏抗体疗效。两种进化的西妥昔单抗变体，Ctx-VY 和 Ctx-Y104D，表现出与 EGFR 的恢复结合能力，EGFR 携带最常见的耐药取代，S492R。Ctx-W52D 与另一个常见的西妥昔单抗耐药取代，G465R（EGFR），具有恢复的结合能力。所有进化的西妥昔单抗变体都能有效地抑制 EGFR 的激活和下游信号转导，并诱导 EGFR 和 EGFR 的内化和降解。在互补决定区有一个或两个氨基酸取代的进化西妥昔单抗变体（Ctx-VY、Ctx-Y104D 和 Ctx-W52D）在很大程度上继承了西妥昔单抗的优化物理和化学性质，从而确保了它们的成药性。我们的数据表明，基于结构的和噬菌体辅助进化是一种有效的、通用的方法，可以逆转受体突变介导的对治疗性抗体药物的耐药性。
