CHR-6494 is a potent and specific haspin inhibitor (antagonist) with IC50 of 2 nM. CHR-6494 inhibits the phosphorylation of histone H3T3. CHR-6494 may be used in cancer-related research.

Physicochemical Properties

Molecular Formula	C16H16N6
Molecular Weight	292.346
Exact Mass	292.143
CAS #	1333377-65-3
Related CAS #	CHR-6494 TFA;1458630-17-5
PubChem CID	70679308
Appearance	Light yellow to khaki solid powder
Density	1.4±0.1 g/cm3
Index of Refraction	1.747
LogP	2.48
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	22
Complexity	378
Defined Atom Stereocenter Count	0
InChi Key	CZZCAOGIEGXMBZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C16H16N6/c1-2-7-17-15-5-6-16-18-10-14(22(16)21-15)11-3-4-13-12(8-11)9-19-20-13/h3-6,8-10H,2,7H2,1H3,(H,17,21)(H,19,20)
Chemical Name	3-(1H-indazol-5-yl)-N-propylimidazo[1,2-b]pyridazin-6-amine
Synonyms	CHR-6494 CHR6494 CHR 6494
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	The growth of dyes such as HCT-116, HeLa, MDA-MB-231, and Wi-38 cells is dose-dependently inhibited by CHR-6494 (0-10-5 nM; 72 hours); the IC50 of 500 nM and 473 nM CHR, respectively, are produced by CHR-6494 (500 nM). Additionally, the spindle assembly checkpoint protein BUB1 and the mitotic division marker cyclin B1, 752 nM and 1059 nM, are upregulated by CHR-6494 (500 nM). With an IC50 ranging from 396 nM to 1229 nM, CHR-6494 exhibits inhibitory effect against melanoma cell lines, including BRAFV600E mutants, NRAS mutants, and wild-type cells [2]. 300 nM and 600 nM of CHR-6494. ; 72 hours) causes a cell exchange that increases the activity of caspase 3/7 by three and six times in COLO-792 cells, and eighty-five and sixteen times in RPMI-7951 cells [2]. When CHR-6494 and MEK dye are combined, they can effectively suppress melanoma cell viability, accelerate melanoma cell shutdown, individually control the course of the cell cycle by stopping melanoma cells at various stages, and stop melanoma cell migration [2]. The anti-proliferative activity of MLN8237 on MDA-MB-231, SKBR3 breast cancer cells is enhanced by CHR-6494 (50, 200 nM; 1 week) [3]. The combination of MLN8237 and CHR-6494 (200 nM; 72 hours) improves the sterility of SKBR3 and MDA-MB-231 cells [3].
ln Vivo	In nude mice of the human colorectal colon, HCT-116 was shown to inhibit tumor growth in CHR-6494 (50 mg/kg; intraperitoneal injection; 2 cycles of 5 consecutive days, 15 d) without significantly changing body weight [1]. In nude mice carrying MDA-MB-231 xenograft tumors, CHR-6494 (20 mg/kg; intraperitoneal injection for 15 consecutive days) has an inhibitory effect on tumor volume and weight when compared with cosmetics [3]. (20 mg/kg; po) for 15 days straight intraperitoneal administration) increases the suppression of tumor weight and volume in vivo by MLN8237 (20 mg/kg; po) [3].
References	[1]. Antitumor activity of a small-molecule inhibitor of the histone kinase Haspin. Oncogene. 2012 Mar 15;31(11):1408-18. [2]. Anti-Melanoma Activities of Haspin Inhibitor CHR-6494 Deployed as a Single Agent or in a Synergistic Combination with MEK Inhibitor. J Cancer. 2017 Aug 25;8(15):2933-2943. [3]. CRISPR/Cas9 screening identifies a kinetochore-microtubule dependent mechanism for Aurora-A inhibitor resistance in breast cancer. Cancer Commun (Lond). 2021 Feb;41(2):121-139.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~171.03 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.55 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: 1 mg/mL (3.42 mM) in 10% DMSO + 90% Corn Oil (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 10.0 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	3.4206 mL	17.1028 mL	34.2056 mL
	5 mM	0.6841 mL	3.4206 mL	6.8411 mL
	10 mM	0.3421 mL	1.7103 mL	3.4206 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.