GSK-461364 (GSK461364; GSK 461364) is a novel, potent, selective, reversible and ATP-competitive small molecule Polo-like kinase 1 (PLK1) inhibitor with potential antitumor activity. It has a 2.2 nM Ki value and inhibits PLK1. There may be antineoplastic activity to GSK-461364. It exhibits a selectivity of over 1000 times against Plk2/3. GSK461364 selectively inhibits Plk1, causing reversible cell arrest at the G1 and G2 stage without apoptosis in normal cells and selective G2/M arrest followed by apoptosis in a variety of tumor cells. The non-ATP-competitive regulation of mitotic spindle function is carried out by the serine/threonine protein kinase Plk1.
Physicochemical Properties
| Molecular Formula | C27H28F3N5O2S | |
| Molecular Weight | 543.6 | |
| Exact Mass | 543.191 | |
| Elemental Analysis | C, 59.66; H, 5.19; F, 10.48; N, 12.88; O, 5.89; S, 5.90 | |
| CAS # | 929095-18-1 | |
| Related CAS # |
|
|
| PubChem CID | 15983966 | |
| Appearance | white solid powder | |
| Density | 1.4±0.1 g/cm3 | |
| Boiling Point | 658.0±65.0 °C at 760 mmHg | |
| Flash Point | 351.7±34.3 °C | |
| Vapour Pressure | 0.0±2.0 mmHg at 25°C | |
| Index of Refraction | 1.645 | |
| LogP | 3.34 | |
| Hydrogen Bond Donor Count | 1 | |
| Hydrogen Bond Acceptor Count | 9 | |
| Rotatable Bond Count | 7 | |
| Heavy Atom Count | 38 | |
| Complexity | 814 | |
| Defined Atom Stereocenter Count | 1 | |
| SMILES | C(N1CCN(C)CC1)C1C=CC2=C(N(C3SC(C(=O)N)=C(O[C@@H](C4C=CC=CC=4C(F)(F)F)C)C=3)C=N2)C=1 |
|
| InChi Key | ZHJGWYRLJUCMRT-QGZVFWFLSA-N | |
| InChi Code | InChI=1S/C27H28F3N5O2S/c1-17(19-5-3-4-6-20(19)27(28,29)30)37-23-14-24(38-25(23)26(31)36)35-16-32-21-8-7-18(13-22(21)35)15-34-11-9-33(2)10-12-34/h3-8,13-14,16-17H,9-12,15H2,1-2H3,(H2,31,36)/t17-/m1/s1 | |
| Chemical Name | 5-[6-[(4-methylpiperazin-1-yl)methyl]benzimidazol-1-yl]-3-[(1R)-1-[2-(trifluoromethyl)phenyl]ethoxy]thiophene-2-carboxamide | |
| Synonyms |
|
|
| 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 (Ki = 2.2 nM) | |
| ln Vitro |
|
|
| ln Vivo |
|
|
| Enzyme Assay | Kinase reactions are carried out using the Z'-Lyte Assay kit (Ser/Thr peptide 16) in a final assay volume of 10 μL. 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 1 mM DTT, 0.01% Brij 35, 0.01 mg/mL casein, 200 μM ATP, 200 μM Polo Box peptide (NH2-MAGPMQS[pT]PLNGAKK-OH), and 6 nM recombinant Plk1 (H6-tev-PLK 1-603) were briefly included in the reactions. For sixty minutes, Plk1 is preincubated with or without 0–1,000 nM GSK461364. Next, a 2 μM peptide is added to start reactions. Reactions are quenched, processed in accordance with the Z'-Lyte protocol, and read on a plate reader after 15 minutes at 23°C. Substratum and product standards are used to convert raw fluorescence values to product concentration. GraFit software is used to perform a two-parameter fit (IC50 and Hill coefficient) in order to determine IC50 values. An upper limit for the Ki*app of GSK461364 is found by applying the Cheng-Prusoff relationship for a competitive inhibitor (ATP Km*app=16 μM) to the IC50 value obtained with 60 minutes of preincubation of GSK461364, as the potency of inhibition for GSK461364 is observed to vary as a function of the ATP concentration in a manner consistent with an ATP-competitive mode of inhibition. | |
| Cell Assay | Cell lines are cultured in the suggested media in a humidified incubator at 37°C and 5% CO2. In triplicate, 1,000 cells are plated in each well of 96-well microtiter plates using culture media. The following day, one plate is harvested with 50 μL of CellTiter-Glo for a time 0 (T=0) measurement, and GSK461364 (GSK461364A) dissolved in DMSO or negative control (0.1% DMSO) is added. | |
| Animal Protocol | Nude mice are used to implant cells, which develop into tumor xenografts. Dosing started at around 100 mm3 for tumors. Every two days (q2d×6, q2d×12) or every four days (q4d×3), mice are given GSK461364 (GSK461364A) or the vehicle [4% DMA/Cremaphore (50:50), pH 5.6] intraperitoneally (i.p.) at nominal dose levels of 25, 50, and 100 mg/kg/dose. For n = 7–8 mice, the results are presented as the median tumor volume. For comparison, paclitaxel (30 mg/kg i.v.; q4d×3) is utilized as a positive control. Vernier calipers are used to measure tumors three times a week. The volume of the tumor is estimated from two-dimensional measurements using the following formula: tumor volume mm3 = (length × width2) × 0.5. The highest dose (approximately 4 g) that results in >20% mortality or >20% weight loss is known as the maximum tolerated dose. Tumor growth delay (TGD), partial regression (PR), or complete regression (CR) are three ways to characterize antitumor activity. The time difference (TGD) between the treated and control tumors to reach a predefined tumor volume (1,000 mm3) is represented. A PR is defined as a tumor's volume decreasing to half of its initial starting volume over a minimum of one week (based on three measurements in a row). A tumor's volume must decrease to less than 13 mm3 for at least one week in order to be considered CR. | |
| References |
[1]. Phase I study of GSK461364, a specific and competitive Polo-like kinase 1 inhibitor, in patients with advanced solid malignancies. Clin Cancer Res. 2011 May 15;17(10):3420-30. [2]. Sensitivity of cancer cells to Plk1 inhibitor GSK461364A is associated with loss of p53 function and chromosome instability. Mol Cancer Ther. 2010 Jul;9(7):2079-89. [3]. Distinct concentration-dependent effects of the polo-like kinase 1-specific inhibitor GSK461364A, including differential effect on apoptosis. Cancer Res. 2009 Sep 1;69(17):6969-77. [4]. Cytotoxic mechanism of PLK1 inhibitor GSK461364 against osteosarcoma: Mitotic arrest, apoptosis, cellular senescence, and synergistic effect with paclitaxel. Int J Oncol. 2016 Mar;48(3):1187-94. |
|
| Additional Infomation |
5-[6-[(4-methyl-1-piperazinyl)methyl]-1-benzimidazolyl]-3-[(1R)-1-[2-(trifluoromethyl)phenyl]ethoxy]-2-thiophenecarboxamide is a member of (trifluoromethyl)benzenes. Polo-like Kinase 1 Inhibitor GSK461364 is a small molecule Polo-like kinase 1 (PLK1) inhibitor with potential antineoplastic activity. Polo-like kinase 1 inhibitor GSK461364 selectively inhibits Plk1, inducing selective G2/M arrest followed by apoptosis in a variety of tumor cells while causing reversible cell arrest at the G1 and G2 stage without apoptosis in normal cells. Plk1, named after the polo gene of Drosophila melanogaster, is a serine/threonine protein kinase involved in regulating mitotic spindle function in a non-ATP competitive manner. |
Solubility Data
| Solubility (In Vitro) |
|
|||
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.60 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 (4.60 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 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 (4.60 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: 1% DMSO +30% polyethylene glycol+1% Tween 80 : 30 mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8396 mL | 9.1979 mL | 18.3959 mL | |
| 5 mM | 0.3679 mL | 1.8396 mL | 3.6792 mL | |
| 10 mM | 0.1840 mL | 0.9198 mL | 1.8396 mL |