AZA1 is a potent inhibitor of Rac1 and Cdc42. AZA1 causes apoptosis and inhibits proliferation, migration and invasion of prostate cancer/tumor cells.

Physicochemical Properties

Molecular Formula	C22H20N6
Molecular Weight	368.434403419495
Exact Mass	368.174
CAS #	1071098-42-4
PubChem CID	25104141
Appearance	Light brown to brown solid powder
LogP	5
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	28
Complexity	529
Defined Atom Stereocenter Count	0
SMILES	N1C(C)=CC2C=C(C=CC1=2)NC1C=CN=C(N=1)NC1C=CC2=C(C=1)C=C(C)N2
InChi Key	SYWHWWKOIJCMKF-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H20N6/c1-13-9-15-11-17(3-5-19(15)24-13)26-21-7-8-23-22(28-21)27-18-4-6-20-16(12-18)10-14(2)25-20/h3-12,24-25H,1-2H3,(H2,23,26,27,28)
Chemical Name	2-N,4-N-bis(2-methyl-1H-indol-5-yl)pyrimidine-2,4-diamine
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

ln Vitro	Human prostate cancer cell 22Rv1 growth is inhibited by AZA1 (Rac1/Cdc42-IN-1) (2-10 μM; 72 hours) [1]. EGF-stimulated 22Rv1 prostate cancer cells exhibit reduced phosphorylation of PAK1, AKT, and BAD upon treatment with AZA1 (2-10 μM; 24 hours) [1]. AZA1 (10 μM; 24 hours) suppresses Rac1- and Cdc42-dependent cell cycle events in 22Rv1 prostate cancer cells [1]. AZA1 inhibits the migration of prostate cancer cells PC-3, DU 145, and 22Rv1 that is dependent on Rac1 and Cdc42 [1]. AZA1 suppresses Rac1 and Cdc42 activity through PAK1/2 phosphorylation, consequently altering cell motility and actin rearrangement of prostate cancer cells [1].
ln Vivo	AZA1 (Rac1/Cdc42-IN-1) (100 μg; i.p.; once daily for two weeks) efficiently suppresses the growth of human 22Rv1 xenografts and enhances mouse survival [1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: 22Rv1 Prostate Cancer Cell Tested Concentrations: 2, 5, 10 μM Incubation Duration: 72 hrs (hours) Experimental Results: Inhibited the proliferation of 22Rv1 Prostate Cancer Cell in unstimulated and EGF-stimulated cancer cells in a dose-dependent manner. Western Blot Analysis[1] Cell Types: 22Rv1 prostate cancer cells (EGF stimulated) Tested Concentrations: 2, 5, 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: diminished phosphorylation of PAK1, AKT and BAD in EGF-stimulated 22Rv1 prostate cancer cells.
Animal Protocol	Animal/Disease Models: 5weeks old athymic nu/nu (nude) mice (carrying 22Rv1 prostate cancer cell xenografts) [1] Doses: 100 μg in 100 µl 30% DMSO one time/day for 2 weeks Experimental Results: AZA1 The inhibitory effect on tumor growth is significant.
References	[1]. A Rac1/Cdc42 GTPase-specific small molecule inhibitor suppresses growth of primary human prostate cancer xenografts and prolongs survival in mice. PLoS One. 2013;8(9):e74924. Published 2013 Sep 11. [2]. Sialylation and glycosylation modulate cell adhesion and invasion to extracellular matrix in human malignant lymphoma: Dependency on integrin and the Rho GTPase family. Int J Oncol. 2015;47(6):2091‐2099.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~135.71 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.79 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 25.0 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: ≥ 2.5 mg/mL (6.79 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.7142 mL	13.5711 mL	27.1422 mL
	5 mM	0.5428 mL	2.7142 mL	5.4284 mL
	10 mM	0.2714 mL	1.3571 mL	2.7142 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.