CiTCoM, UMR CNRS 8038, U1268 INSERM, Université Paris Cité, Faculté de Santé, UFR de Pharmacie, 4 av. de l'Observatoire, 75006 Paris, France.
CiTCoM, UMR CNRS 8038, Université Paris Cité, Faculté de Santé, UFR de Pharmacie, 4 av. de l'Observatoire, 75006 Paris, France.
J Med Chem. 2023 Jul 27;66(14):9753-9765. doi: 10.1021/acs.jmedchem.3c00561. Epub 2023 Jul 6.
Macromolecular ligands targeting vascular endothelial growth factor A (VEGF) to inhibit pathological angiogenesis are used in the clinic for the treatment of cancers and ocular diseases. To develop smaller ligands retaining high affinity through an avidity effect, here we design homodimer peptides targeting the two symmetrical binding sites of the VEGF homodimer. A series of 11 dimers were synthesized with flexible poly(ethylene glycol) (PEG) linkers of increasing lengths. The binding mode was determined by size exclusion chromatography, and analytical thermodynamic parameters were measured by isothermal titration calorimetry and compared to the antibody bevacizumab. The effect of linker length was qualitatively correlated to a theoretical model. With the optimal length in PEG-dimer , the binding affinity was improved 40-fold compared to a monomer control, resulting in a single-digit nanomolar value. Finally, we validated the benefit of the dimerization strategy by evaluating the activity of control monomers and selected dimers in cell-based assays with human umbilical vein endothelial cells (HUVECs).
针对血管内皮生长因子 A（VEGF）的高分子配体可用于抑制病理性血管生成，临床上可用于治疗癌症和眼部疾病。为了开发通过亲合力效应保留高亲和力的更小配体，我们设计了针对 VEGF 同源二聚体两个对称结合位点的同源二聚体肽。用一系列不同长度的柔性聚乙二醇（PEG）接头合成了 11 个二聚体。通过排阻色谱确定了结合模式，并通过等温滴定微量热法测量了分析热力学参数，并与抗体 bevacizumab 进行了比较。接头长度的影响与理论模型定性相关。在 PEG-二聚体的最佳长度下，与单体对照相比，结合亲和力提高了 40 倍，导致数值达到纳摩尔级。最后，我们通过在基于细胞的人脐静脉内皮细胞（HUVEC）测定中评估对照单体和选定二聚体的活性，验证了二聚化策略的益处。
