SP-96 is a specific and ATP-competitive Aurora B inhibitor (antagonist) with IC50 of 0.316 nM and >2000-fold selectivity for FLT3 and KIT. SP-96 displays selective growth inhibition in NCI60 screening and is selective for breast cancer cells such as MDA-MD-468 (GI50=107 nM). SP-96 may be utilized in research related to triple-negative breast cancer (TNBC).

Physicochemical Properties

Molecular Formula	C25H20FN7O
Molecular Weight	453.47
Exact Mass	453.171
CAS #	2682114-54-9
PubChem CID	154573766
Appearance	White to off-white solid powder
LogP	4
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	5
Heavy Atom Count	34
Complexity	684
Defined Atom Stereocenter Count	0
SMILES	C1(NC(=O)NC2=CC=CC(NC3=NC=NC4C3=CC=C(C=4)C3C=NN(C)C=3)=C2)=CC(=CC=C1)F
InChi Key	CGDYRZUPKPBLND-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H20FN7O/c1-33-14-17(13-29-33)16-8-9-22-23(10-16)27-15-28-24(22)30-20-6-3-7-21(12-20)32-25(34)31-19-5-2-4-18(26)11-19/h2-15H,1H3,(H,27,28,30)(H2,31,32,34)
Chemical Name	1-(3-fluorophenyl)-3-[3-[[7-(1-methylpyrazol-4-yl)quinazolin-4-yl]amino]phenyl]urea
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	Aurora A 18.975 nM (IC50) Aurora B 0.316 nM (IC50)
ln Vitro	SP-96 is a very strong, focused, and non-ATP competitive inhibitor of Aurora B (IC50=0.316 nM) that exhibits >2000 fold selectivity against KIT (IC50=1307.6 nM) and FLT3 (IC50=1475.6 nM) [1]. While SP-96 (0-1 μM; 24 hours) is not promiscuous, it is selective for certain cell lines, as evidenced by its GI50 values of 107 nM, 47.4 nM, 50.3 nM, and 53.2 nM for MDA-MB-468, CCRF-CEM, COLO 205, and A498 cells, respectively[1]. By raising the DNA content and causing the cell volume and nucleus to enlarge, SP-96 (63.2 nM) suppresses the Aurora B activity of H460 cells [1]. The enzyme activity of Aurora B is inhibited by SP-96 (0-2 μM) at an IC50 of 0.316 nM, while Aurora A enzyme activity is inhibited at an IC50 of 18.975 nM. More than 2000-fold selectivity is shown by SP-96 for both KIT (IC50=1307.6 nM) and FLT3 (IC50=1475.6 nM). Moreover, it has an IC50 value of less than 2 μM and inhibits the activity of EGFR, RET, and HER2 receptor tyrosine kinases (RTKs) [1].
Cell Assay	Cell Viability Assay[1] Cell Types: MDA-MB-468, CCRF-CEM, COLO 205 and A498 cell Tested Concentrations: 0 -1 µM Incubation Duration: 24 hrs (hours) Experimental Results: demonstrated good inhibitory activity on MDA-MB-468 cells.
References	[1]. Discovery of SP-96, the first non-ATP-competitive Aurora Kinase B inhibitor, for reduced myelosuppression.Eur J Med Chem. 2020 Jul 12;203:112589.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (220.52 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.51 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 25.0 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	2.2052 mL	11.0261 mL	22.0522 mL
	5 mM	0.4410 mL	2.2052 mL	4.4104 mL
	10 mM	0.2205 mL	1.1026 mL	2.2052 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.