ENMD-2076 (ENMD2076) is a novel,potent and orally bioactive inhibitor of multikinase including Aurora A and Flt3 with potential antitumor activity. It inhibits Aurora A and Flt3 with IC50s of 14 nM and 1.86 nM, respectively, and exhibits 25-fold selectivity for Aurora A over Aurora B. It shows potent in vitro antiproliferative activity and high in vivo antitumor efficacy.

Physicochemical Properties

Molecular Formula	C21H25N7
Molecular Weight	375.470103025436
Exact Mass	375.217
CAS #	934353-76-1
Related CAS #	ENMD-2076 Tartrate;1453868-32-0
PubChem CID	16041424
Appearance	White to light yellow solid powder
Density	1.3±0.1 g/cm3
Boiling Point	535.0±50.0 °C at 760 mmHg
Flash Point	277.4±30.1 °C
Vapour Pressure	0.0±1.4 mmHg at 25°C
Index of Refraction	1.704
LogP	1.31
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	5
Heavy Atom Count	28
Complexity	499
Defined Atom Stereocenter Count	0
SMILES	CC1=CC(=NN1)NC2=CC(=NC(=N2)/C=C/C3=CC=CC=C3)N4CCN(CC4)C
InChi Key	BLQYVHBZHAISJM-CMDGGOBGSA-N
InChi Code	InChI=1S/C21H25N7/c1-16-14-20(26-25-16)23-19-15-21(28-12-10-27(2)11-13-28)24-18(22-19)9-8-17-6-4-3-5-7-17/h3-9,14-15H,10-13H2,1-2H3,(H2,22,23,24,25,26)/b9-8+
Chemical Name	(E)-N-(5-methyl-1H-pyrazol-3-yl)-6-(4-methylpiperazin-1-yl)-2-styrylpyrimidin-4-amine.
Synonyms	ENMD 2076; ENMD-2076; ENMD2076.
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	ENMD-2076 is specific for Aurora A over Aurora B (IC50=350 nM). ENMD-2076 inhibits HUVEC proliferation at an IC50 of 0.15 mM. The IC50 values for 10 human leukemia cell lines range from 0.025 to 0.53 mM. In this panel, MV4:11 cells are the most responsive by a factor of more than 4. The lymphoma-derived U937 cell line treated with ENMD-2076 demonstrates a dose-dependent increase in G2-M-phase arrest and apoptosis. ENMD-2076 suppresses cellular Flt3 ligand (FL)-induced Flt3 autophosphorylation in THP-1 cells, which have been demonstrated to express FL-responsive wild-type Flt-3 (18), with an IC50 of 28 nM. ENMD-2076 inhibits stem cell factor (SCF)-induced Kit autophosphorylation in MO7e cells with an IC50 of 40 nM. ENMD-2076 inhibits VEGFR2/KDR autophosphorylation with an IC50 of 7 nM [1].
ln Vivo	Treatment with ENMD-2076 leads to statistically significant tumor growth or regression inhibition that is dose dependent. Furthermore, as fast-growing tumors like A375 melanoma and slow-growing tumors like HT29 colon carcinoma are similarly inhibited by ENMD-2076, there is no correlation between tumor growth rate and antitumor efficacy, which is theoretically expected for a mitotic kinase inhibitor. With the exception of the A375 model, studies have not found any weight loss or indications of morbidity at daily doses of up to 302 mg/kg (equivalent to 200 mg/kg of the free base) for ENMD-2076.
Animal Protocol	Dissolved in water or ENMD-2076 free base in CMC-Tween vehicle (0.075% carboxymethylcellulose, 0.085% Tween 80 in water); 300 mg/kg; Oral gavage Tumor models including HCT-116, HT29, CT-26, A375, MDA-MB-231, H929, OPM-2, MV4;11 and HL60 are established in CB.17 SCID or NCr nude mice.
References	[1]. ENMD-2076 is an orally active kinase inhibitor with antiangiogenic and antiproliferative mechanisms of action. Mol Cancer Ther. 2011 Jan;10(1):126-37. [2]. Preclinical activity of a novel multiple tyrosine kinase and aurora kinase inhibitor, ENMD-2076, against multiple myeloma. Br J Haematol. 2010 Aug;150(3):313-25.
Additional Infomation	Aurora A Kinase/Tyrosine Kinase Inhibitor ENMD-2076 is an orally bioavailable synthetic small molecule with potential antiangiogenic and antineoplastic activities. Aurora A kinase/tyrosine kinase inhibitor ENMD-2076 selectively binds to and inhibits non-specified tyrosine kinases and Aurora kinases (AKs). The inhibition of AKs may result in the inhibition of cell division and proliferation and may induce apoptosis in tumor cells that overexpress AKs; antiangiogenic activity is related to the inhibition of angiogenic tyrosine kinases. AKs are serine-threonine kinases that play an essential role in mitotic checkpoint control during mitosis and are important regulators of cell division and proliferation.

Solubility Data

Solubility (In Vitro)

DMSO: 105 mg/mL (279.6 mM) Water: 1 mg/mL (2.7 mM) Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.66 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 (6.66 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 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (6.66 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.. Solubility in Formulation 4: 0.5% CMC+0.25% Tween 80:30mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.6633 mL	13.3166 mL	26.6333 mL
	5 mM	0.5327 mL	2.6633 mL	5.3267 mL
	10 mM	0.2663 mL	1.3317 mL	2.6633 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.