Discovery of Covalent Inhibitors Targeting the Transcriptional Enhanced Associate Domain Central Pocket

Hacer Karatas,Sasikala Thavam,Herbert Waldmann,Wanjin Hong,Ajaybabu V Pobbati,Axel Pahl,Hélène Adihou,Alexandra Friese,Congbao Kang,Christine Nowak,Sarah Zinken,Stéphanie M Guéret,Lara Dötsch,Malte Metz,Gernot Hahne,Elizabeth Yihui Ng,Sonja Sievers,Mohammad Akbarzadeh

doi:10.1021/acs.jmedchem.0c01275

Abstract

Transcriptional enhanced associate domain (TEAD) transcription factors together with coactivators and corepressors modulate the expression of genes that regulate fundamental processes, such as organogenesis and cell growth, and elevated TEAD activity is associated with tumorigenesis. Hence, novel modulators of TEAD and methods for their identification are in high demand. We describe the development of a new “thiol conjugation assay” for identification of novel small molecules that bind to the TEAD central pocket. The assay monitors prevention of covalent binding of a fluorescence turn-on probe to a cysteine in the central pocket by small molecules. Screening of a collection of compounds revealed kojic acid analogues as TEAD inhibitors, which covalently target the cysteine in the central pocket, block the interaction with coactivator yes-associated protein with nanomolar apparent IC50 values, and reduce TEAD target gene expression. This methodology promises to enable new medicinal chemistry programs aimed at the modulation of TEAD activity.

Highlights

Transcriptional enhanced associate domain (TEAD) transcription factors play significant roles during development, tissue regeneration, cell growth, and migration and in human disease progression.1−4 Elevated TEAD expression has been observed in solid tumors such as prostate,5 lung,6 colorectal,7 gastric,8 and breast cancers.9 A number of TEAD target genes including cell surface receptor tyrosine kinase Axl,10 connective tissue growth factor (CTGF),11,12 apoptosis inhibitor survivin,13 and tumor marker mesothelin14 are frequently associated with tumorigenesis
TEAD1 is involved in cardiogenesis, while TEAD2 is critical for neural development and TEAD4 is necessary for embryo implantation.15−18 pharmacological modulation of TEAD activity may enable novel medicinal chemistry approaches both in oncology and regenerative medicine
We have developed a new “thiol conjugation assay” that enables the systematic identification of small-molecule inhibitors of the TEAD central pocket

Summary

■ INTRODUCTION

Transcriptional enhanced associate domain (TEAD) transcription factors play significant roles during development, tissue regeneration, cell growth, and migration and in human disease progression.− Elevated TEAD expression has been observed in solid tumors such as prostate, lung, colorectal, gastric, and breast cancers. A number of TEAD target genes including cell surface receptor tyrosine kinase Axl, connective tissue growth factor (CTGF), apoptosis inhibitor survivin, and tumor marker mesothelin are frequently associated with tumorigenesis. Nonsteroidal anti-inflammatory drugs flufenamic acid (1, Kd = 73 μM) and niflumic acid (2, Kd = 28 μM) bind to the TEAD central pocket and inhibit expression of TEAD− YAP target genes and growth of cancer cell lines.. A chloromethylketone analogue of flufenamic acid (3, TED-347) was developed as a covalent TEAD inhibitor that can allosterically inhibit the TEAD−YAP interaction (Ki = 10.4 μM).. We describe the development of a new methodology for the identification of novel small-molecule inhibitors binding to the TEAD central pocket. By means of a newly developed “thiol conjugation assay” we identified kojic acid derivatives that covalently target the palmitoylatable cysteine in the TEAD central pocket and inhibit the TEAD−YAP interaction with nanomolar apparent IC50 values. Synthesis of Kojic Acid Analoguesa aReagents and conditions: (a) EtOH, rt, 2−20 days. (b) MeI, K2CO3, DMF, 24 h, rt

■ RESULTS AND DISCUSSION

■ CONCLUSIONS

M hydroxylamine solution

■ ACKNOWLEDGMENTS

■ REFERENCES