AZ304 is a dual BRAF inhibitor that effectively inhibits wild-type BRAF, mutant BRAF (V600E) and wild-type CRAF with IC50s of 79 nM, 38 nM and 68 nM respectively. AZ304 has significant inhibitory effects on other kinases, such as p38 (IC50, 6 nM), CSF1R (IC50, 35 nM). Has anti-tumor activity.

Physicochemical Properties

Molecular Formula	C27H25N5O2
Exact Mass	451.2
Elemental Analysis	C, 71.82; H, 5.58; N, 15.51; O, 7.09
CAS #	942507-42-8
PubChem CID	16202218
Appearance	White to off-white solid powder
LogP	5.2
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	6
Heavy Atom Count	34
Complexity	749
Defined Atom Stereocenter Count	0
InChi Key	NGWQZRWVYYFTHC-UHFFFAOYSA-N
InChi Code	InChI=1S/C27H25N5O2/c1-17-8-9-20(31-26(33)18-6-5-7-19(12-18)27(2,3)15-28)13-23(17)32-25-22-11-10-21(34-4)14-24(22)29-16-30-25/h5-14,16H,1-4H3,(H,31,33)(H,29,30,32)
Chemical Name	3-(2-cyanopropan-2-yl)-N-(3-((7-methoxyquinazolin-4-yl)amino)-4-methylphenyl)benzamide
Synonyms	AZ304; AZ-304; AZ 304;
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	BRafV600E 38 nM (IC50) BRAFWT 79 nM (IC50) CRAF 68 nM (IC50) p38 6 nM (IC50) CSF1R 35 nM (IC50) MAP3K7 6400 nM (IC50) CSK 7050 nM (IC50)
ln Vitro	AZ304 (1 nM–100 μM) significantly lowers p-ERK phosphorylation; in the V600E mutant BRAF-containing melanoma cell line A375, the mean EC50 was 65 nM, whereas in the wild type BRAF melanoma cell line SK-MEL-31, both with and without EGF, the EC50s were 52 nM and 60 nM[1]. In both BRAF genetic status cell lines, AZ304 also strongly suppresses p-p38[1]. With GI50s of 4.539 μM, 3.896 μM, 4.987 μM, 1.763 μM (48 hours) and 0.5032 μM, 0.3887 μM, 0.6354 μM, 0.3772 μM (72 hours), respectively, AZ304 (0, 0.1, 1, 10, 100 μM, 48 and 72 hours) dose-dependently suppresses the growth of RKO, HT-29, DiFi, and Caco-2[1]. AZ304 (2 μM, 36 and 48 hours) raises p-EGFR in BRAF wild type and BRAF V600E mutant cells while decreasing BRAF, p-ERK, p-AKT, and p-mTOR levels. When combined with C225, AZ304 more effectively reduces the BRAF, ERK, AKT, and mTOR signaling pathways while downregulating p-EGFR and inhibiting p-ERK[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: RKO, HT-29, DiFi, Caco-2 cells Tested Concentrations: 0, 0.1, 1, 10, 100 μM Incubation Duration: 48, 72 hrs (hours) Experimental Results: Dose- dependently inhibited the growth of V600E mutant BRAF cell lines (RKO, HT-29) and wild-type BRAF cell lines (DiFi, Caco-2).
References	[1]. AZ304, a novel dual BRAF inhibitor, exerts anti-tumour effects in colorectal cancer independently of BRAF genetic status. Br J Cancer. 2018 May;118(11):1453-1463.

Solubility Data

Solubility (In Vitro)

DMSO : 125 mg/mL (276.84 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.61 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.08 mg/mL (4.61 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.08 mg/mL (4.61 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)