TAK-960 (TAK960) is a novel, investigational, orally bioavailable, and selective inhibitor of polo-like kinase 1 (PLK1) with anticancer activity. TAK-960 also exhibits inhibitory activities against PLK2 and PLK3, with IC50s of 16.9 and 50.2 nM, respectively. It inhibits PLK1 with an IC50 of 0.8 nM at 10 μM ATP.Serine/threonine protein kinase PLK1 is essential for several processes in mitosis. Human PLK1 is a promising target for anticancer therapy because it has been demonstrated to be overexpressed in a variety of human cancers and because elevated levels of PLK1 have been linked to a poor prognosis. Multiple tumor cell lines, including those that express multidrug-resistant protein 1 (MDR1), have demonstrated TAK-960's efficacy. TAK-960 treatment resulted in G(2)-M cell accumulation, abnormal polo mitosis morphology, and elevated phosphorylation of histone H3 (pHH3) in vitro and in vivo, all of which were consistent with PLK1 inhibition. TAK-960 did not affect nondividing normal cells (EC(50) >1,000 nmol/L), but it did inhibit the proliferation of several cancer cell lines, with mean EC(50) values ranging from 8.4 to 46.9 nmol/L. In the tested cell lines, there was no correlation found between the potency of TAK-960 and the mutation status of TP53, KRAS, and MDR1 expression. Oral administration of TAK-960 in animal models resulted in a dose-dependent increase in pHH3 and a significant inhibition of the growth of HT-29 colorectal cancer xenografts. The administration of TAK-960 once daily demonstrated noteworthy effectiveness against various tumor xenograft models, such as a disseminated leukemia model and an adriamycin/paclitaxel-resistant xenograft model. TAK-960 is currently being evaluated clinically in patients with metastatic cancer.
Physicochemical Properties
| Molecular Formula | C27H36CL2F3N7O3 |
| Molecular Weight | 561.59916 |
| Exact Mass | 561.267 |
| Elemental Analysis | C, 57.74; H, 6.10; F, 10.15; N, 17.46; O, 8.55 |
| CAS # | 1137868-52-0 |
| Related CAS # | TAK-960 dihydrochloride;TAK-960 hydrochloride;1137868-96-2;TAK-960 monohydrochloride;2108449-45-0 |
| PubChem CID | 53357478 |
| Appearance | White to beige solid powder |
| Density | 1.4±0.1 g/cm3 |
| Index of Refraction | 1.615 |
| LogP | 1.33 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 11 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 40 |
| Complexity | 903 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | 0 |
| InChi Key | GWRSATNRNFYMDI-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C27H34F3N7O3/c1-35-10-8-16(9-11-35)32-24(38)18-12-22(40-3)20(13-19(18)28)33-26-31-14-21-23(34-26)37(17-6-4-5-7-17)15-27(29,30)25(39)36(21)2/h12-14,16-17H,4-11,15H2,1-3H3,(H,32,38)(H,31,33,34) |
| Chemical Name | 4-[(9-cyclopentyl-7,7-difluoro-5-methyl-6-oxo-8H-pyrimido[4,5-b][1,4]diazepin-2-yl)amino]-2-fluoro-5-methoxy-N-(1-methylpiperidin-4-yl)benzamide |
| Synonyms | TAK 960; TAK960; TAK-960 |
| 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 | PLK1 (IC50 = 0.8 nM); PLK2 (IC50 = 16.9 nM); PLK3 (IC50 = 50.2 nM); FAK/PTK2 (IC50 = 19.6 nM); MLCK/MYLK (IC50 = 25.6 nM); FES/FPS (IC50 = 58.2 nM) |
| ln Vitro | TAK-960 exhibited activity in a number of tumor cell lines, including those that express MDR1, the multidrug resistance protein. (Source: ) In line with PLK1 inhibition, TAK-960 treatment results in an increase in phosphorylation of histone H3 (pHH3), an accumulation of G2/M cells, and aberrant "polo" mitosis morphology. (Source: ) TAK-960 does not affect non-dividing normal cells (EC50 >1,000 nM), but it does inhibit the proliferation of several cancer cell lines, with mean EC50 values ranging from 8.4 to 46.9 nM. In the tested cell lines, there is no correlation between the potency of TAK-960 and the mutation status of TP53, KRAS, or MDR1 expression.[1] |
| ln Vivo | Oral TAK-960 administration in animal models causes a dose-dependent increase in pHH3 and a significant inhibition of the growth of HT-29 colorectal cancer xenografts.[1] Once-daily TAK-960 treatment shows notable efficacy against a variety of tumor xenografts, such as a disseminated leukemia model and a xenograft model resistant to doxorubicin and paritamol.[1] |
| Enzyme Assay | The TR-FRET assay, which measures the ATP-dependent phosphorylation of a biotinylated substrate peptide corresponding to residues 2470 through 2488 of the mammalian target of rapamycin protein (Biotin-AGAGTVPESIHSFIGDGLV), is used to evaluate the inhibitory activity of TAK-960. Using HotSpotSM technology, 288 kinases are screened for TAK-960 inhibition (1 μM), and the IC50 values of the kinases that pass the test are found. |
| Cell Assay | In 96-well plates, cells are seeded at a density of 3–30 × 103 cells/well using the suitable medium plus 10% fetal calf serum (FCS). The number of viable cells is determined using the CellTiter-Glo Assay 72 hours after the cells are treated with serial dilutions of TAK-960 for 24 hours. Statistical analysis and EC50 value computation are carried out. |
| Animal Protocol |
Athymic nude mice (BALB/cAJc1-nu/nu), severe combined immunodeficiency (SCID) mice (C.B17-Icr- scid/scid Jcl) or NOD-scid mice (NOD.CB17-Prkdc scid/J) 30 mg/kg Oral dosing |
| References |
[1]. TAK-960, a novel, orally available, selective inhibitor of polo-like kinase 1, shows broad-spectrum preclinical antitumor activity in multiple dosing regimens. Mol Cancer Ther. 2012 Mar;11(3):700-9. [2]. PLK1 blockade enhances therapeutic effects of radiation by inducing cell cycle arrest at the mitotic phase. Sci Rep. 2015 Oct 27;5:15666. |
| Additional Infomation | PLK1 Inhibitor TAK-960 is an orally available, Polo-like kinase 1 (PLK1) inhibitor with potential antineoplastic activity. Polo-like kinase 1 inhibitor TAK-960 selectively inhibits PLK1, inducing selective G2/M cell-cycle arrest followed by apoptosis in a variety of tumor cells while causing reversible cell-cycle arrest at the G1 and G2 stages without apoptosis in normal cells. PLK1 inhibition may result in the inhibition of proliferation in PLK1-overexpressed tumor cells. PLK1, named after the polo gene of Drosophila melanogaster, is a serine/threonine kinase crucial in the regulation of mitosis. |
Solubility Data
| Solubility (In Vitro) |
DMSO: 13~16.7 mg/mL (23.2~29.7 mM) Ethanol: ~3 mg/mL (~5.3 mM) |
| 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 | 1.7806 mL | 8.9031 mL | 17.8063 mL | |
| 5 mM | 0.3561 mL | 1.7806 mL | 3.5613 mL | |
| 10 mM | 0.1781 mL | 0.8903 mL | 1.7806 mL |