Abstract Colorectal cancer (CRC) remains the second cause of death by cancer worldwide and its survival is currently estimated at 60%1. There is therefore a crucial need to identify biomarkers and new targets for therapy. In 90% of CRC, the Wnt/β-catenin pathway is constitutively activated due to mutations of β-catenin, APC (Adenomatous Polyposis Coli) or axin-12. These alterations cause an accumulation of β-catenin and lead to an over-transcription of target genes involved in tumorigenesis3-4. Studies of our team pointed out an overexpression of the Protein Tyrosine Kinase 7 (PTK7) in CRC, an event associated with metastatic development, reduced metastases-free survival, and resistance to chemotherapy. In CRC cells, the pseudokinase PTK7 has pro-metastatic and pro-migratory functions, and thus appears to be a promising new therapeutic target5-6. The role of PTK7 in the Wnt/β-catenin pathway has been demonstrated in several studies7-8. Our group identified β-catenin as a partner of PTK7. We selected a first series of small molecules inhibitors targeting PTK7/β-catenin interaction9. This strategy could represent, in the future, a new therapeutic strategy to inhibit CRC cell growth dependent on the Wnt signaling pathway. However, the compounds identified had weak activity, with an IC50 of around 10-25 mM. We have set up a new method with SYNSIGHT to select more potent chemical inhibitors targeting PTK7/β-catenin in CRC in order to counteract the Wnt/β-catenin signaling pathway deregulation.