Description: Hesperadin is a novel, potent and ATP-competitive inhibitor of human aurora B kinase with potential antitumor activity. It inhibits Aurora B with IC50 of 250 nM in a cell-free assay. It markedly reduces the activity of AMPK, Lck, MKK1, MAPKAP-K1, CHK1 and PHK while it does not inhibit MKK1 activity in vivo. Hersperadin shows potent in vitro antiproliferative activity and high in vivo antitumor efficacy.

Physicochemical Properties

Molecular Formula	C29H32N4O3S
Molecular Weight	516.65
Exact Mass	516.219
CAS #	422513-13-1
Related CAS #	Hesperadin hydrochloride
PubChem CID	135421442
Appearance	Light yellow to yellow solid powder
Density	1.3±0.1 g/cm3
Index of Refraction	1.675
LogP	3.13
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	8
Heavy Atom Count	37
Complexity	855
Defined Atom Stereocenter Count	0
InChi Key	GLDSKRNGVVYJAB-DQSJHHFOSA-N
InChi Code	InChI=1S/C29H32N4O3S/c1-2-37(35,36)32-24-15-16-26-25(19-24)27(29(34)31-26)28(22-9-5-3-6-10-22)30-23-13-11-21(12-14-23)20-33-17-7-4-8-18-33/h3,5-6,9-16,19,30,32H,2,4,7-8,17-18,20H2,1H3,(H,31,34)/b28-27-
Chemical Name	(Z)-N-(2-oxo-3-(phenyl((4-(piperidin-1-ylmethyl)phenyl)amino)methylene)indolin-5-yl)ethanesulfonamide
Synonyms	Hesperadine; Hesperadin; Hesperadine;
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	Hesperadin (10-100 nM) suppresses Aurora kinase-1 (TbAUK1)-mediated phosphorylation of trypanosome histone H3 (TbH3) in a dose-dependent manner, with an IC50 of 40 nM [1]. Hesperadin (0.01-10 μM; 24 or 48 hours) reduces the growth of bloodstream (BF) and procyclic (PF) cultures [1]. Hesperadin (100-200 nM; 24-72 hours) affects cell shape and hinders cell cycle progression, similar to RNAi suppression of TbAUK1 [1].
ln Vivo	Hesperadin (20 mg/kg/d; IV) works in concert with temozolomide (TMZ) to extend the survival of xenograft mice[2].
Cell Assay	Cell Viability Assay[1] Cell Types: M110 cells Tested Concentrations: 0.01, 0.1, 1, 10 μM Incubation Duration: 24 hrs (hours) or 48 hrs (hours) Experimental Results: Inhibiting growth of BF cultures with IC50 of 50 nM, while the inhibition of PF growth required approximately 11-fold more Hesperadin, with IC50 of 550 nM. Cell Cycle Analysis[1] Cell Types: M110 cells Tested Concentrations: 100, 200 nM Incubation Duration: 24, 48, 72 hrs (hours) Experimental Results: Had a strong effect on cell growth and mitotic progression at 100-200 nM.
Animal Protocol	Animal/Disease Models: 6weeks old female nude mice injected GBM cells[2] Doses: 20 mg /kg/d Route of Administration: Iv injection Experimental Results: Increased the survival of xenograft mice models.
References	[1]. The cell cycle as a therapeutic target against Trypanosoma brucei: Hesperadin inhibits Aurora kinase-1 and blocks mitotic progression in bloodstream forms. Mol Microbiol. 2009 Apr; 72(2): 442-58. [2]. Targeting Aurora kinase B attenuates chemoresistance in glioblastoma via a synergistic manner with temozolomide. Pathol Res Pract. 2019 Nov; 215(11): 152617. [3]. Chemical Genomics Approach Leads to the Identification of Hesperadin, an Aurora B Kinase Inhibitor, as a Broad-Spectrum Influenza Antiviral. Int J Mol Sci. 2017 Sep 8;18(9):1929.
Additional Infomation	Hesperadin is an oxindole that is indolin-2-one which is substituted at position 5 by an (ethylsulfonyl)nitrilo group and at position 2 by a methylidene group, which is itself substituted by a phenyl group and a [4-(piperidin-1-ylmethyl)phenyl]amino group. An Aurora B kinase inhibitor, it is used to inhibit chromosome alignment and segregation. It has a role as an Aurora kinase inhibitor. It is a member of oxindoles, a member of piperidines, a sulfonamide, an enamine and a tertiary amino compound.

Solubility Data

Solubility (In Vitro)

DMSO: 103 mg/mL (199.4 mM) Water:<1 mg/mL Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.84 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 (4.84 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 3: 30% PEG400+0.5% Tween80+5% propylene glycol: 30 mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9355 mL	9.6777 mL	19.3555 mL
	5 mM	0.3871 mL	1.9355 mL	3.8711 mL
	10 mM	0.1936 mL	0.9678 mL	1.9355 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.