CW069 (CW-069; CW 069) is a potent, selective and allosteric inhibitor of microtubule motor protein HSET with potential antitumor activity. It inhibits HSET with an IC50 of 75 μM.

Physicochemical Properties

Molecular Formula	C23H21IN2O3
Molecular Weight	500.33
Exact Mass	500.059
CAS #	1594094-64-0
Related CAS #	1594094-64-0
PubChem CID	73427517
Appearance	White to off-white solid powder
Density	1.6±0.1 g/cm3
Boiling Point	666.0±55.0 °C at 760 mmHg
Flash Point	356.6±31.5 °C
Vapour Pressure	0.0±2.1 mmHg at 25°C
Index of Refraction	1.687
LogP	6.74
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	8
Heavy Atom Count	29
Complexity	532
Defined Atom Stereocenter Count	1
SMILES	C1=CC=C(C=C1)C[C@@H](C(=O)NC2=C(C=C(C=C2)I)C(=O)O)NCC3=CC=CC=C3
InChi Key	IRDIXDXDSUBHIU-NRFANRHFSA-N
InChi Code	InChI=1S/C23H21IN2O3/c24-18-11-12-20(19(14-18)23(28)29)26-22(27)21(13-16-7-3-1-4-8-16)25-15-17-9-5-2-6-10-17/h1-12,14,21,25H,13,15H2,(H,26,27)(H,28,29)/t21-/m0/s1
Chemical Name	5-Iodo-2-[[(2S)-1-oxo-3-phenyl-2-[(phenylmethyl)amino]propyl]amino]-benzoic acid
Synonyms	CW069; CW-069; CW 069
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	CW-069 is an HSET allosteric inhibitor, with an IC50 of 75 μM. Selectivity over KSP that is statistically significant is demonstrated by CW-069. CW-069 exhibits a robust suppressive effect on N1E-115 cells and a less potent inhibitory effect on NHDF cells, with IC50 values of 86 ± 10 μM and 181 ± 7 μM, respectively. When applied in N1E-115 cells with supernumerary centrosomes, CW-069 (100 or 200 μM) increases multipolar spindles, but it has no effect on the shape of bipolar spindles in normal human dermal fibroblast cells. While CW-069 (200 μM) inhibits monastrol's suppression of KSP and causes multipolar anaphase and cell death in N1E-115 cells transfected with HSET siRNA, it does not cause mitotic arrest in HeLa cells[1].
ln Vivo
Animal Protocol
References	[1]. Design, synthesis, and biological evaluation of an allosteric inhibitor of HSET that targets cancer cells with supernumerary centrosomes. Chem Biol. 2013 Nov 21;20(11):1399-410.

Solubility Data

Solubility (In Vitro)

DMSO:100 mg/mL (199.9 mM) Water:<1 mg/mL Ethanol:4 mg/mL (8.0 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.16 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 (4.16 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 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 (4.16 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.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9987 mL	9.9934 mL	19.9868 mL
	5 mM	0.3997 mL	1.9987 mL	3.9974 mL
	10 mM	0.1999 mL	0.9993 mL	1.9987 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.