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疾患原因タンパク質を分解するUボディによる新規創薬戦略

研究課題

研究課題/領域番号 23KF0197
研究種目

特別研究員奨励費

配分区分基金
応募区分外国
審査区分 小区分37030:ケミカルバイオロジー関連
研究機関東京大学

研究代表者

菅 裕明  東京大学, 大学院理学系研究科(理学部), 教授 (00361668)

研究分担者 PHILIPPE GREGOIRE  東京大学, 大学院理学系研究科(理学部), 外国人特別研究員
研究期間 (年度) 2023-11-15 – 2026-03-31
研究課題ステータス 交付 (2023年度)
配分額 *注記
2,000千円 (直接経費: 2,000千円)
2025年度: 500千円 (直接経費: 500千円)
2024年度: 1,000千円 (直接経費: 1,000千円)
2023年度: 500千円 (直接経費: 500千円)
キーワードUbiquitin / cancer therapy / proteasomal degradation
研究開始時の研究の概要

The aim of this project is to develop a new type of molecule that relies on ubiquitin grafting to selectively degrade targeted proteins. 1) Proof of concept using a well characterized protein 2) Application to other targets and cell delivery 3) Publication/ patent & blood-brain barrier targeting

研究実績の概要

- The ability to ubiquitinate a protein using Ubodies (modified ubiquitin) was confirmed.
- Targeting MDM2, an inhibitor of tumor suppressor p53, often requires a helical conformation that mimics the structure of the p53 binding site. New types of selection were developed to improve the helical propensity of the modified loop in Ubodies:
-A) Fully randomized sequence of 8 to 15 amino acid (control)
-B) Randomized sequence (8-15 amino acids) with helix inducers (3-5) on both sides (sequence of amino acids that will help attain the desired structure)
-C) Grafted a sequence known to bind to the target (pDI) and optimized the flanking region (1 to 5 amino acids on both sides) using AlphaFold. The amino acids of the flanking regions are randomized for the selection.

現在までの達成度 (区分)
現在までの達成度 (区分)

3: やや遅れている

理由

The first selection ((B) helix inducers + randomized sequence) did not show high recovery rate (ratio of protein binding to the beads / total mRNA in the selection). After sequencing, 20 proteins were recombinantly expressed but none of them showed binding to the protein target (MDM2). After optimization, the recovery rate was improved by 100 times and the selection strategy was diversified (as explained in the summary of achievements) to increase the chances of finding binding candidates. A competition selection between (A) and (B) was also performed to attempt to scale binding affinity against ease of translation (D). A delivery method using lipid extracted from cells and manufactured into liposomes is being developed in parallel and will be used once a MDM2 binder Ubody is found.

今後の研究の推進方策

The four selection schemes (A-D) will be sequenced, and ~5 binders / selection will be selected for recombinant expression. After binders are found, delivery into cancer cells will be performed using lipofectamine and the reconstituted liposomes. The impact on normal and cancer cell growth will be assessed, and the mechanism of action carefully monitored to conclude on whether the activity comes from protein-protein interaction inhibition (activation of p53 lead to an increase in MDM2 levels due to a negative feedback loop), and/ or degradation of MDM2 by the proteasome (level of MDM2 stay constant or decrease during the assay). Application to other targets will then be performed.

報告書

(1件)
  • 2023 実施状況報告書
  • 研究成果

    (1件)

すべて 2024

すべて 雑誌論文 (1件) (うち国際共著 1件、 査読あり 1件)

  • [雑誌論文] Delivery to, and Reactivation of, the p53 Pathway in Cancer Cells Using a Grafted Cyclotide Conjugated with a Cell-Penetrating Peptide2024

    • 著者名/発表者名
      Philippe Gregoire Jean-Baptiste、Huang Yen-Hua、Mittermeier Anna、Brown Christopher J.、Kaas Quentin、Ramlan Siti Radhiah、Wang Conan K.、Lane David、Loewer Alexander、Troeira Henriques S?nia、Craik David J.
    • 雑誌名

      Journal of Medicinal Chemistry

      巻: 67 号: 2 ページ: 1197-1208

    • DOI

      10.1021/acs.jmedchem.3c01682

    • 関連する報告書
      2023 実施状況報告書
    • 査読あり / 国際共著

URL: 

公開日: 2023-11-17   更新日: 2024-12-25  

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