Information AZ304 is a synthetic inhibitor designed to interact with the ATP-binding site ofwild type and V600E mutant BRAFwith IC50 values of 79 nM and 38 nM, respectively. It also inhibitsCRAF, p38 and CSF1Rat sub 100 nM potencies. Targets p38 (Cell-free assay); CSF1R (Cell-free assay); BRAF(V600E) (Cell-free assay); CRAF (Cell-free assay); WT BRAF (Cell-free assay) 6 nM; 35 nM; 38 nM; 68 nM; 79 nM In vitro AZ304 shows potent inhibitory activities to the kinase domains of wild type BRAF, V600E mutant BRAF and wild type CRAF with IC50 values of 79\u2009nM, 38\u2009nM and 68\u2009nM, respectively. AZ304 potently reduces ERK phosphorylation (p-ERK), with a mean EC50 of 65\u2009nM in the V600E mutant BRAF containing melanoma cell line A375 and an EC50 of 60\u2009nM in the wild type BRAF containing melanoma cell line SK-MEL-31. AZ304 markedly inhibits cell proliferation in mutant BRAF cancer cell lines, and effectively reduces cell growth in selected cell lines harbouring wild type BRAF/RAS or mutant RAS. The GI50 values ranged from 0.08-7.72\u2009μM in mutant BRAF cell lines, 0.43-11.7\u2009μM in wild type BRAF/RAS cell lines, and 0.9-16.66\u2009μM in mutant RAS cell lines. AZ304 exhibits anti-proliferative effects on multiple cancer types, including melanoma, colorectal cancer, leukaemia, ovarian cancer, lung cancer, and pancreatic cancer, independently of BRAF genetic status. AZ304 retains inhibitory activity against both V600E mutant and wild type BRAF CRC cell lines in the presence of the EGFR ligand EGF. In vivo AZ304 monotherapy and its combination with Cetuximab have anti-tumour effects on RKO and Caco-2 tumour xenografts without obvious toxicity, independently of BRAF mutation status. Cell Research(from reference) Cell lines：cell lines containing wild type BRAF or V600E mutant BRAF (MC-F7, A549 and A375 cells) Concentrations：6.5 nM, 65 nM, 650 nM and 6.5 μM Incubation Time：75 min