Piperlongumine conjugates induce targeted protein degradation

Jing Pei; Yufeng Xiao; Xingui Liu; Wanyi Hu; Amin Sobh; Yaxia Yuan; Shuo Zhou; Nan Hua; Samuel G Mackintosh; Xuan Zhang; Kari B Basso; Manasi Kamat; Qingping Yang; Jonathan D Licht; Guangrong Zheng; Daohong Zhou; Dongwen Lv

doi:10.1016/j.chembiol.2023.01.004

Piperlongumine conjugates induce targeted protein degradation

Cell Chem Biol. 2023 Feb 16;30(2):203-213.e17. doi: 10.1016/j.chembiol.2023.01.004. Epub 2023 Feb 6.

Authors

Jing Pei¹, Yufeng Xiao², Xingui Liu¹, Wanyi Hu², Amin Sobh³, Yaxia Yuan¹, Shuo Zhou¹, Nan Hua¹, Samuel G Mackintosh⁴, Xuan Zhang², Kari B Basso⁵, Manasi Kamat⁵, Qingping Yang¹, Jonathan D Licht³, Guangrong Zheng⁶, Daohong Zhou⁷, Dongwen Lv⁸

Affiliations

¹ Department of Pharmacodynamics, College of Pharmacy, University of Florida, 1333 Center Drive, Gainesville, FL 32610, USA.
² Department of Medicinal Chemistry, College of Pharmacy, University of Florida, 1333 Center Drive, Gainesville, FL 32610, USA.
³ Division of Hematology/Oncology, University of Florida Health Cancer Center, 2033 Mowry Road, Suite 145, Gainesville, FL 32610, USA.
⁴ Department of Biochemistry and Molecular Biology, College of Medicine, University of Arkansas for Medical Sciences, 4301 W. Markham Street, Slot 803, Little Rock, AR 72205, USA.
⁵ Department of Chemistry, University of Florida, PO Box 117200, Gainesville, FL 32611, USA.
⁶ Department of Medicinal Chemistry, College of Pharmacy, University of Florida, 1333 Center Drive, Gainesville, FL 32610, USA. Electronic address: zhengg@cop.ufl.edu.
⁷ Department of Pharmacodynamics, College of Pharmacy, University of Florida, 1333 Center Drive, Gainesville, FL 32610, USA; Department of Biochemistry and Structural Biology and Center for Innovative Drug Discovery, School of Medicine, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA; Mays Cancer Center, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA. Electronic address: zhoud@uthscsa.edu.
⁸ Department of Pharmacodynamics, College of Pharmacy, University of Florida, 1333 Center Drive, Gainesville, FL 32610, USA; Department of Biochemistry and Structural Biology and Center for Innovative Drug Discovery, School of Medicine, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA; Mays Cancer Center, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA. Electronic address: lyud@uthscsa.edu.

Abstract

Proteolysis targeting chimeras (PROTACs) are bifunctional molecules that degrade target proteins through recruiting E3 ligases. However, their application is limited in part because few E3 ligases can be recruited by known E3 ligase ligands. In this study, we identified piperlongumine (PL), a natural product, as a covalent E3 ligase recruiter, which induces CDK9 degradation when it is conjugated with SNS-032, a CDK9 inhibitor. The lead conjugate 955 can potently degrade CDK9 in a ubiquitin-proteasome-dependent manner and is much more potent than SNS-032 against various tumor cells in vitro. Mechanistically, we identified KEAP1 as the E3 ligase recruited by 955 to degrade CDK9 through a TurboID-based proteomics study, which was further confirmed by KEAP1 knockout and the nanoBRET ternary complex formation assay. In addition, PL-ceritinib conjugate can degrade EML4-ALK fusion oncoprotein, suggesting that PL may have a broader application as a covalent E3 ligase ligand in targeted protein degradation.

Keywords: CDK9; E3 ligase; KEAP1; PROTAC; TurboID; piperlongumine.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Kelch-Like ECH-Associated Protein 1 / metabolism
Ligands
NF-E2-Related Factor 2* / metabolism
Proteolysis
Ubiquitin-Protein Ligases* / metabolism

Substances

N-(5-(((5-(1,1-dimethylethyl)-2-oxazolyl)methyl)thio)-2-thiazolyl)-4-piperidinecarboxamide
Kelch-Like ECH-Associated Protein 1
piperlongumine
NF-E2-Related Factor 2
Ubiquitin-Protein Ligases
Ligands

Abstract

Publication types

MeSH terms

Substances

Grants and funding