Wang Lei, Qiao Yu, Zong Huifang, Han Lei, Ke Yong, Pan ZhiDi, Chen Jie, Lu Jun, Li Jinyao, Ying Tianlei, Zhang Baohong, Zhu Jianwei
Engineering Research Center of Cell and Therapeutic Antibody, MOE, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
Jecho Institute, Co. Ltd., Shanghai, China.
Front Pharmacol. 2022 Feb 23;13:803059. doi: 10.3389/fphar.2022.803059. eCollection 2022.
Colorectal cancer is a commonly diagnosed cancer with high mortality worldwide. Postoperative recidivation and metastasis still are the main challenges in clinical treatments. Thus, it is urgent to develop new therapies against colorectal cancer. Epithelial Cell Adhesion Molecule (EpCAM) is overexpressed in colorectal cancer cells and strongly associated with cancer development. Bispecific antibody (BsAb) is a kind of promising immunotherapy, which could recognize T cells and cancer cells simultaneously to achieve the anti-tumor effects. A bispecific antibody targeting EpCAM and CD3 with IgG format was genereated by split intein based on the Bispecific Antibody by Protein Splicing" platform. , the affinity of CD3×EpCAM BsAb was determined by Biolayer interferometry, its cytotoxicity was detected by LDH release assay, T cell recruitment and activation was detected by Flow Cytometry. , its pharmacokinetic parameters were detected, and anti-tumor effects were evaluated on the tumor cell xenograft mouse model. The results showed that the CD3×EpCAM BsAb could activate and recruit T cells via binding colorectal cells and T cells, which could lead to more potent cytotoxicity to various colorectal cell lines than its parent EpCAM monoclonal antibody (mAb) . The CD3×EpCAM BsAb had similar pharmacokinetic parameters with EpCAM mAb and inhibits tumor growth on the SW480 tumor cell xenograft mouse model. The CD3×EpCAM BsAb could be a promising candidate for colorectal cancer treatment.
结直肠癌是一种在全球范围内诊断率高且死亡率高的癌症。术后复发和转移仍然是临床治疗中的主要挑战。因此，开发针对结直肠癌的新疗法迫在眉睫。上皮细胞粘附分子（EpCAM）在结直肠癌细胞中过度表达，且与癌症发展密切相关。双特异性抗体（BsAb）是一种有前景的免疫疗法，它可以同时识别T细胞和癌细胞以实现抗肿瘤效果。基于“蛋白质剪接双特异性抗体”平台，通过分裂内含肽产生了一种IgG形式的靶向EpCAM和CD3的双特异性抗体。通过生物膜干涉术测定CD3×EpCAM BsAb的亲和力，通过乳酸脱氢酶释放试验检测其细胞毒性，通过流式细胞术检测T细胞募集和激活。检测其药代动力学参数，并在肿瘤细胞异种移植小鼠模型上评估抗肿瘤效果。结果表明，CD3×EpCAM BsAb可通过结合结直肠细胞和T细胞激活并募集T细胞，这比其亲本EpCAM单克隆抗体（mAb）对各种结直肠细胞系具有更强的细胞毒性。CD3×EpCAM BsAb与EpCAM mAb具有相似的药代动力学参数，并在SW480肿瘤细胞异种移植小鼠模型上抑制肿瘤生长。CD3×EpCAM BsAb可能是结直肠癌治疗的一个有前景的候选药物。
