Rational design of a sensitivity-enhanced tracer for discovering efficient APC-Asef inhibitors

Jie Zhong; Yuegui Guo; Shaoyong Lu; Kun Song; Ying Wang; Li Feng; Zhen Zheng; Qiufen Zhang; Jiacheng Wei; Peng Sang; Yan Shi; Jianfeng Cai; Guoqiang Chen; Chen-Ying Liu; Xiuyan Yang; Jian Zhang

doi:10.1038/s41467-022-32612-6

Rational design of a sensitivity-enhanced tracer for discovering efficient APC-Asef inhibitors

Nat Commun. 2022 Aug 24;13(1):4961. doi: 10.1038/s41467-022-32612-6.

Authors

Jie Zhong^#^{1

2

3}, Yuegui Guo^#⁴, Shaoyong Lu^#², Kun Song², Ying Wang², Li Feng³, Zhen Zheng³, Qiufen Zhang³, Jiacheng Wei², Peng Sang⁵, Yan Shi⁵, Jianfeng Cai⁵, Guoqiang Chen⁶, Chen-Ying Liu⁷, Xiuyan Yang^{8

9}, Jian Zhang^{10

11

12

13}

Affiliations

¹ State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine at Shanghai, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
² Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
³ Medicinal Chemistry and Bioinformatics Center, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁴ Department of Colorectal and Anal Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁵ Department of Chemistry, University of South Florida, Tampa, FL, USA.
⁶ Research Unit of Stress and Cancer, Chinese Academy of Medical Sciences, Shanghai, China.
⁷ Department of Colorectal and Anal Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China. liuchenying@xinhuamed.com.cn.
⁸ Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, China. yangxy@shsmu.edu.cn.
⁹ State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau, China. yangxy@shsmu.edu.cn.
¹⁰ State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine at Shanghai, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China. jian.zhang@sjtu.edu.cn.
¹¹ Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, China. jian.zhang@sjtu.edu.cn.
¹² Medicinal Chemistry and Bioinformatics Center, Shanghai Jiao Tong University School of Medicine, Shanghai, China. jian.zhang@sjtu.edu.cn.
¹³ School of Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, China. jian.zhang@sjtu.edu.cn.

^# Contributed equally.

Abstract

The adenomatous polyposis coli (APC)-Rho guanine nucleotide exchange factor 4 (Asef) protein-protein interaction (PPI) is essential for colorectal cancer metastasis, making it a promising drug target. Herein, we obtain a sensitivity-enhanced tracer (tracer 7) with a high binding affinity (K_d = 0.078 μM) and wide signal dynamic range (span = 251 mp). By using tracer 7 in fluorescence-polarization assays for APC-Asef inhibitor screening, we discover a best-in-class inhibitor, MAI-516, with an IC₅₀ of 0.041 ± 0.004 μM and a conjugated transcriptional transactivating sequence for generating cell-permeable MAIT-516. MAIT-516 inhibits CRC cell migration by specifically hindering the APC-Asef PPI. Furthermore, MAIT-516 exhibits no cytotoxic effects on normal intestinal epithelial cell and colorectal cancer cell growth. Overall, we develop a sensitivity-enhanced tracer for fluorescence polarization assays, which is used for the precise quantification of high-activity APC-Asef inhibitors, thereby providing insight into PPI drug development.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenomatous Polyposis Coli Protein / genetics
Adenomatous Polyposis Coli Protein / metabolism
Adenomatous Polyposis Coli* / metabolism
Cell Movement
Colorectal Neoplasms* / pathology
Humans
Rho Guanine Nucleotide Exchange Factors / metabolism

Substances

Adenomatous Polyposis Coli Protein
Rho Guanine Nucleotide Exchange Factors