Physicochemical Properties
| Molecular Formula | C18H17F3N6O2 |
| Molecular Weight | 406.361793279648 |
| Exact Mass | 406.136 |
| CAS # | 1458630-17-5 |
| Related CAS # | CHR-6494;1333377-65-3 |
| PubChem CID | 91691123 |
| Appearance | Typically exists as solid at room temperature |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 9 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 29 |
| Complexity | 461 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | FC(C(=O)O)(F)F.N12C(C=CC(NCCC)=N1)=NC=C2C1C=CC2=C(C=NN2)C=1 |
| InChi Key | ILWYDZNXJQESDI-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C16H16N6.C2HF3O2/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;3-2(4,5)1(6)7/h3-6,8-10H,2,7H2,1H3,(H,17,21)(H,19,20);(H,6,7) |
| Chemical Name | 3-(1H-indazol-5-yl)-N-propylimidazo[1,2-b]pyridazin-6-amine;2,2,2-trifluoroacetic acid |
| 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 | haspin 2 nM (IC50) |
| ln Vitro | With IC50s of 500 nM, 473 nM, 752 nM, and 1059 nM, respectively, CHR-6494 (TFA; 0-105 nM; 72 hours) dose-dependently suppresses the proliferation of cancer cells, including HCT-116, HeLa, MDA-MB-231, and Wi-38 cells[1]. CHR-6494 (TFA; 500 nM) upregulates the spindle assembly checkpoint protein BUB1 and the marker of mitotic arrest cyclin B1, causing a mitotic catastrophe with aberrant spindle shape and centrosome amplification[1]. Angiogenesis is inhibited by CHR-6494 (TFA; 0, 0.5, 1.0 μM; 24 to 36 h) in the ex vivo chicken embryo aortic arch ring assay[1]. With IC50s ranging from 396 nM to 1229 nM, CHR-6494 (TFA) demonstrates inhibitory activity against melanoma cell lines, including BRAFV600E mutants, NRAS mutants, and wild type cells[2]. Inducing apoptosis, CHR-6494 (TFA; 300 nM and 600 nM; 72 hours) boosts caspase 3/7 activity in COLO-792 cells by 3 and 6 times, respectively, and in RPMI-7951 melanoma cells by 8.5 and 16 times, respectively[2]. In MDA-MB-231 and SKBR3 breast cancer cells, CHR-6494 (TFA; 50, 200 nM; 1 week) strengthens the antiproliferative effects of MLN8237[3]. When coupled with MLN8237, CHR-6494 (TFA; 200 nM; 72 hours) increases the apoptosis of MDA-MB-231 and SKBR3 cells[3]. |
| ln Vivo | In nude mice containing HCT-116 human colorectal cancer cells, CHR-6494 (TFA; 50 mg/kg; ip in two cycles of five consecutive days for 15 days) inhibits the growth of tumors[1]. In nude mice with MDA-MB-231 xenograft tumors, CHR-6494 (TFA; 20 mg/kg; intraperitoneal injection for 15 consecutive days) inhibits the growth of the tumor[3]. |
| Animal Protocol |
Animal/Disease Models: Male 4-5 weeks old athymic nu/nu (nude) mice harboring HCT-116 cells xenograft tumor with a tumor volume of 200 mm3[1] Doses: 50 mg/kg (diluted in a solution of 10% DMSO/20% 2-hydroxypropyl- b-cyclodextrin) Route of Administration: ip in two cycles of five days for 15 days Experimental Results: Dose-dependent tumor growth inhibition was demonstrated. Did not change the body weight of mice. Animal/Disease Models: 4weeks old female nude mice bearing MDA -MB-231 xenograft tumors Doses: 20 mg/kg in a final formulation in 10% DMSO/20% 2-hydroxypropyl-β-cyclodextrin Route of Administration: ip for 15 days Experimental Results: Inhibited the tumor volume and weight compared with the control group in nude mice bearing MDA-MB-231 xenograft tumors. Enhanced the tumor volume and weight inhibition of MLN8237 (20 mg/kg; po) in vivo. |
| 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) | May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples |
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4609 mL | 12.3044 mL | 24.6087 mL | |
| 5 mM | 0.4922 mL | 2.4609 mL | 4.9217 mL | |
| 10 mM | 0.2461 mL | 1.2304 mL | 2.4609 mL |