# A synthetic benzoxazine dimer derivative targets c‐Myc to inhibit colorectal cancer progression

## TL;DR

A new compound called ECD targets the c-Myc protein to stop colorectal cancer growth by causing DNA damage and cell death.

## Contribution

ECD is a novel synthetic benzoxazine dimer derivative that inhibits c-Myc by disrupting its interaction with MAX and inducing DNA damage.

## Key findings

- ECD disrupts c-Myc/MAX complexes, leading to c-Myc degradation in colorectal cancer cells.
- ECD induces DNA damage and apoptosis in CRC cells, confirmed by proteomic analysis.
- In vivo studies show ECD significantly inhibits tumor growth in a chick embryo model.

## Abstract

The c‐Myc protein is a well‐known oncoprotein that plays a crucial role in regulating cell growth, proliferation, and differentiation. The overexpression or dysregulation of c‐Myc is commonly associated with tumorigenesis in several cancers, including colorectal cancer (CRC). c‐Myc forms a heterodimer with its partner MAX to activate the expression of various genes. Here, we synthesized a novel c‐Myc‐targeting small molecule, 2,2′‐((cyclohexylazanedyl)bis(methylene))bis(4‐ethylphenol), or ECD, and demonstrate ECD's anticancer activity via interference with the c‐Myc/MAX dimer to promote c‐Myc degradation in CRC cells in vitro, in silico, and in vivo. This study revealed the activity of ECD toward CRC cells as a c‐Myc inhibitor. Computer‐aided analysis revealed that the effect of ECD was mediated through disturbance of the c‐Myc/MAX complex. Moreover, ECD exhibited cytotoxic activity by inducing DNA damage, leading to apoptotic cell death. This DNA damage‐inducing property was also confirmed by whole‐proteome profiling of HT29 cells after ECD treatment. In the chick embryo chorioallantoic membrane (CAM) xenograft assay, we demonstrated a remarkable inhibition of the tumorigenic activity upon ECD exposure. In summary, we identified ECD as a novel potent compound targeting the oncoprotein c‐Myc that may offer new opportunities for CRC treatment.

Benzoxazine dimer derivatives bind to the bHLH‐LZ region of c‐Myc, disrupting c‐Myc/MAX complexes, which are evaluated from SAR analysis. This increases ubiquitination and reduces cellular c‐Myc. Impairing DNA repair mechanisms is shown through proteomic analysis. In vitro and in ovo studies demonstrate tumorigenesis inhibition through cell cycle arrest and apoptosis.