Cell‐Active, Irreversible Covalent Inhibitors Targeting a Surface‐Exposed Non‐Catalytic Lysine on Aurora a Kinase by Using Squarate Chemistry

Z Zuqin Wang (State Key Laboratory of Bioactive Molecules and Druggability Assessment, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education, School of Pharmacy Jinan University #855 Xingye Avenue Guangzhou 510632 China) X Xuan Wang Y Yong Li P Peishan Li S Shengjie Huang (State Key Laboratory of Fluorine and Nitrogen Chemistry and Advanced Materials, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences Chinese Academy of Sciences Shanghai P. R. China) P Peng Chen G Guanghui Tang (Department of Chemistry, National University of Singapore 4 Science Drive 2 Singapore 117543 Singapore) X Xiaotong Ding (State Key Laboratory of Bioactive Molecules and Druggability Assessment, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education, School of Pharmacy Jinan University #855 Xingye Avenue Guangzhou 510632 China) Z Zhang Zhang (State Key Laboratory of Bioactive Molecules and Druggability Assessment, and School of Pharmacy, Jinan University, 601 Huangpu Avenue West, Guangzhou 510632, China) Z Zhi‐Min Zhang (State Key Laboratory of Bioactive Molecules and Druggability Assessment, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education, School of Pharmacy Jinan University #855 Xingye Avenue Guangzhou 510632 China) Y Yang Zhou S Shao Q. Yao (Department of Chemistry, National University of Singapore, 4 Science Drive 2, Singapore 117544, Singapore) X Xiaoyun Lu (School of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou International Campus, Guangzhou 511442, China)

Abstract

AbstractTargeting surface‐exposed lysines in kinases through covalent modification presents a major challenge due to their high pKa and inherently low reactivity. While current research primarily targets more reactive catalytic lysines buried in the ATP‐binding pocket, no systematic rational strategy has yet been developed for selectively engaging surface‐exposed lysines. Herein, we present a versatile strategy for developing cell‐active covalent kinase inhibitors (CKIs) by selectively targeting unique surface‐exposed lysines using squarate chemistry. By using Aurora A (AURKA) as a proof‐of‐concept, we systematically evaluated this approach against other well‐known lysine‐reactive warheads (e.g., EBA, SO2F, and OSO2F) and demonstrated, for the first time, squarates’ superior efficacy in engaging these challenging low‐reactivity lysines. Amongst various AURKA CKIs, AL8 emerged as the first‐in‐class squarate‐based, cell‐active inhibitor, exhibiting excellent selectivity in both biochemical and cellular assays with long‐residence time in covalently engaging endogenous AURKA. Detailed investigation of effects of leaving groups on squaric esters provided valuable insights for future development of lysine‐reactive CKIs. Our finding has established squarate‐containing ligands as a unique and readily tunable platform for covalent modification of surface‐exposed, non‐catalytic lysines in targeted kinase drug discovery.

Article Details

Volume / Issue Vol. 64, Issue 40
Published September 26, 2025
ISSN 1433-7851
Publisher Wiley

Journal Info

Angewandte Chemie International Edition

Wiley

ISSN: 1433-7851 Physical Sciences

Authors (13)

Z

Zuqin Wang

State Key Laboratory of Bioactive Molecules and Druggability Assessment, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education, School of Pharmacy Jinan University #855 Xingye Avenue Guangzhou 510632 China

X

Xuan Wang

Y

Yong Li

P

Peishan Li

S

Shengjie Huang

State Key Laboratory of Fluorine and Nitrogen Chemistry and Advanced Materials, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences Chinese Academy of Sciences Shanghai P. R. China

P

Peng Chen

G

Guanghui Tang

Department of Chemistry, National University of Singapore 4 Science Drive 2 Singapore 117543 Singapore

X

Xiaotong Ding

State Key Laboratory of Bioactive Molecules and Druggability Assessment, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education, School of Pharmacy Jinan University #855 Xingye Avenue Guangzhou 510632 China

Z

Zhang Zhang

State Key Laboratory of Bioactive Molecules and Druggability Assessment, and School of Pharmacy, Jinan University, 601 Huangpu Avenue West, Guangzhou 510632, China

Z

Zhi‐Min Zhang

State Key Laboratory of Bioactive Molecules and Druggability Assessment, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education, School of Pharmacy Jinan University #855 Xingye Avenue Guangzhou 510632 China

Y

Yang Zhou

S

Shao Q. Yao

Department of Chemistry, National University of Singapore, 4 Science Drive 2, Singapore 117544, Singapore

X

Xiaoyun Lu

School of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou International Campus, Guangzhou 511442, China