2. Academic Validation
  3. A practical "preTACs-cytoblot" platform accelerates the streamlined development of PROTAC-based protein degraders

A practical "preTACs-cytoblot" platform accelerates the streamlined development of PROTAC-based protein degraders

  • Eur J Med Chem. 2023 May 5:251:115248. doi: 10.1016/j.ejmech.2023.115248.
Zijian Rao 1,  Kailin Li 1,  Ju Hong 1,  Danni Chen 1,  Baoli Ding 1,  Li Jiang 1,  Xuxin Qi 1,  Jiawen Hu 1,  Bo Yang 2,  Qiaojun He 3,  Xiaowu Dong 4,  Ji Cao 5,  Cheng-Liang Zhu 6
Affiliations

Affiliations

  • 1 Institute of Pharmacology & Toxicology, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, PR China.
  • 2 Institute of Pharmacology & Toxicology, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, PR China; Innovation Institute for Artificial Intelligence in Medicine, Zhejiang University, Hangzhou, 310016, PR China; Hangzhou Institute of Innovative Medicine, Zhejiang University, Hangzhou, 310058, PR China.
  • 3 Institute of Pharmacology & Toxicology, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, PR China; Centre for Drug Safety Evaluation and Research of ZJU, Hangzhou, 310058, PR China; Innovation Institute for Artificial Intelligence in Medicine, Zhejiang University, Hangzhou, 310016, PR China; Hangzhou Institute of Innovative Medicine, Zhejiang University, Hangzhou, 310058, PR China; Cancer Centre, Zhejiang University, Hangzhou, 310058, PR China.
  • 4 Institute of Pharmacology & Toxicology, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, PR China; Innovation Institute for Artificial Intelligence in Medicine, Zhejiang University, Hangzhou, 310016, PR China; Hangzhou Institute of Innovative Medicine, Zhejiang University, Hangzhou, 310058, PR China; Cancer Centre, Zhejiang University, Hangzhou, 310058, PR China.
  • 5 Institute of Pharmacology & Toxicology, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, PR China; Innovation Institute for Artificial Intelligence in Medicine, Zhejiang University, Hangzhou, 310016, PR China; Hangzhou Institute of Innovative Medicine, Zhejiang University, Hangzhou, 310058, PR China; Cancer Centre, Zhejiang University, Hangzhou, 310058, PR China; Engineering Research Center of Innovative Anticancer Drugs, Ministry of Education, China. Electronic address: [email protected].
  • 6 Institute of Pharmacology & Toxicology, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, PR China; Centre for Drug Safety Evaluation and Research of ZJU, Hangzhou, 310058, PR China; Innovation Institute for Artificial Intelligence in Medicine, Zhejiang University, Hangzhou, 310016, PR China; Hangzhou Institute of Innovative Medicine, Zhejiang University, Hangzhou, 310058, PR China; Engineering Research Center of Innovative Anticancer Drugs, Ministry of Education, China. Electronic address: [email protected].
PMID: 36905918 DOI: 10.1016/j.ejmech.2023.115248
Abstract

With the growing importance of PROTAC-mediated protein degradation in drug discovery, robust synthetic methodologies and rapid screening assays are urgently needed. By harnessing the improved alkene hydroazidation reaction, we developed a novel strategy to introduce azido groups into the linker-E3 ligand conjugates and effectively created a range of prepacked terminal azide-labeled "preTACs" as PROTAC toolkit building blocks. Moreover, we demonstrated that preTACs are ready to conjugate to ligands targeting a protein of interest to generate libraries of chimeric degraders, which are subsequently screened for effective protein degradation directly from cultured cells with a cytoblot assay. Our study exemplifies that this practical "preTACs-cytoblot" platform allows efficient PROTAC assembly and rapid activity assessments. It may help industrial and academic investigators to accelerate their streamlined development of PROTAC-based protein degraders.

Keywords

Click chemistry; Cytoblot; PROTAC; Protein degrader; Staudinger reaction.

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