抗KRAS環状ペプチドファマコフォアの小型タンパク質へのラソグラフト体の創出

将抗 KRAS 环肽药效基团移植到小蛋白质上

• 批准号: 22KF0089
• 负责人: 菅 裕明
• 金额: $ 1.47万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2023
• 资助国家: 日本
• 起止时间: 2023-03-08 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22KF0089/
• 关键词: KRas; lasso-graft; macrocylic peptides; RaPID

Two anti-KRas peptides (KD2 and KWT9) have been grafted to Ubiquitin to provide two initial generations of hybrid constructs called “Ubodies” with different selectivity profiles. KD2-derived Ubodies are selective for the KRas “on” vs “off” state. KWT9 is newly identified, and displays (together with its Ubody derivatives) selectivity towards the mutant KRas over the healthy protein. A further generation of de novo Ubodies was also developed. Initial evaluation shows several hit compounds featuring excellent activity on the malignant target protein and selectivity over the healthy protein and/or the “off” state. Early results from collaborators validate the potential of the peptides, although further experimental work is needed on the Ubodies. The overall workflow for Ubody development has now been streamlined, from peptide hit identification, lasso-graft tether optimization, expression and purification to binding kinetics estimation. This experimental scheme can now be applied to different targets. The main challenge remaining to this project is now the delivery of the Ubodies into the cell by viral vector transfection.

两种抗Kras Pepperides（KD2和KWT9）已嫁接到泛素，提供了具有不同选择性概况的两代初始的混合构建体。 KD2衍生的Ubodies对Kras“ ON”与“ OFF”状态具有选择性。 KWT9是新鉴定的，并且（及其UBODER衍生物）对突变体Kras对健康蛋白的选择性具有选择性。还开发了进一步的从头造物。初始评估显示，几种命中化合物具有在恶性靶蛋白上具有出色活性以及对健康蛋白质和/或“ OFF”状态的选择性的出色活性。尽管在Ubodies上需要进一步的实验性工作，但合作者的早期结果证明了Petides的潜力。现在，从胡椒命中率识别，套索绑带的束缚优化，表达和纯化到结合动力学估计的整体工作流程已经简化了。现在，该实验方案可以应用于不同的目标。该项目剩下的主要挑战是通过病毒矢量翻译将Ubodies传递到细胞中。