Structure guided maturation of a novel humanized anti-HBV antibody and its preclinical development
Structure guided maturation of a novel humanized anti-HBV antibody and its preclinical development
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新型人源化抗乙型肝炎抗体的结构引导成熟及其临床前开发
DOI：
10.1016/j.antiviral.2020.104757
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Xia N
中科院分区：
文献类型：
--
作者：
Zhou B;Xia L;Zhang T;You M;Huang Y;He M;Su R;Tang J;Zhang J;Li S;An Z;Yuan Q;Luo W;Xia N
我们已经报道，E6 F6，一种小鼠单克隆抗体，是一种有前途的治疗慢性B型肝炎（CH B）患者的选择。通过CDR移植法获得亲和力丧失的人源化E6 F6抗体B11。为了解决这个问题，在B11：HBsAg的预测免疫复合物模型上，使用CHARMM力场方法通过扫描诱变进行计算机模拟亲和力成熟。我们选择了四种具有最高增加的相互作用能的变体用于进一步表征。在两个点突变（重链：Asp 65 Val; His 66 Leu）内的抗体huE 6 F6 -1被鉴定为恢复亲本抗体的高结合亲和力、中和活性和体内病毒抑制的有效功效。基于晶体结构（1.8 Å分辨率）的分子对接证明huE 6 F6 - 1中形成了更稳定、更紧密的氢键相互作用。电子显微镜观察到huE 6 F6 -1的HBV免疫复合物更小、更分散，表明它将具有与亲代E6 F6 mAb相同的治疗功效。huE 6 F6 -1的临床前研究和药代动力学表明，它是一个稳定的和理想的领导候选人，以改善慢性乙型肝炎的临床管理。值得注意的是，我们的结构引导方法可以在药物开发期间促进其他啮齿动物抗体候选物的人源化和亲和力成熟。
We have reported that E6F6, a mouse monoclonal antibody, is a promising treatment option for patients with chronic hepatitis B (CHB). A humanized E6F6 antibody B11 with affinity loss was obtained by CDR-grafting approach. To address this issue,in silicoaffinity maturation through scanning mutagenesis using CHARMM force field methods was performed on an predicted immune complex model of the B11:HBsAg. We chose four variants with top increased interaction energy for further characterization. The antibody huE6F6-1 within two point mutations (Heavy Chain: Asp65Val; His66Leu) was identified to restore the parental antibody's high binding affinity, neutralization activity, and potent efficacy of viral suppressionin vivo. Crystal structure (1.8 Å resolution) based molecular docking proved more stabilized and compact hydrogen bond interactions formed in huE6F6-1.The smaller and dispersed HBV immune complexes of huE6F6-1 by electron microscopy suggested it will have the same therapeutic efficacy as the parental E6F6 mAb. Preclinical study and pharmacokinetics of huE6F6-1 demonstrated that it is a stable and desirable lead candidate to improve the clinical management of CHB. Notably, our structure guided approach may facilitate the humanization and affinity maturation of other rodent antibody candidates during drug development.