GW843682X is a novel, potent, selective, ATP-competitive inhibitor of polo-like kinase 1 (PLK1) and polo-like kinase 3 (PLK3) with IC50s of 2.2 nM and 9.1 nM, respectively. It exhibits selectivity against approximately 30 other kinases that is greater than 100-fold. GW843682X caused 5-8F cells to undergo apoptosis and inhibited their proliferation in a dose-dependent manner (IC50=62.5-125nmol/L). By down-regulating the expression of the IAP gene, GW843682X demonstrated remarkable cytotoxic effects on nasopharyngeal carcinoma 5-8F cells. This suggests that GW843682X may develop into a novel therapeutic agent for nasopharyngeal carcinoma.
Physicochemical Properties
| Molecular Formula | C22H18N3O4F3S |
| Molecular Weight | 477.45622 |
| Exact Mass | 477.10 |
| Elemental Analysis | C, 55.34; H, 3.80; F, 11.94; N, 8.80; O, 13.40; S, 6.71 |
| CAS # | 660868-91-7 |
| Related CAS # | 660868-91-7 |
| Appearance | Light yellow to yellow solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 629.6±65.0 °C at 760 mmHg |
| Flash Point | 334.6±34.3 °C |
| Vapour Pressure | 0.0±1.8 mmHg at 25°C |
| Index of Refraction | 1.613 |
| LogP | 4.25 |
| SMILES | O=C(C1=C(OCC2=CC=CC=C2C(F)(F)F)C=C(N3C4=CC(OC)=C(OC)C=C4N=C3)S1)N |
| InChi Key | JSKUWFIZUALZLX-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C22H18F3N3O4S/c1-30-16-7-14-15(8-17(16)31-2)28(11-27-14)19-9-18(20(33-19)21(26)29)32-10-12-5-3-4-6-13(12)22(23,24)25/h3-9,11H,10H2,1-2H3,(H2,26,29) |
| Chemical Name | 5-(5,6-dimethoxybenzimidazol-1-yl)-3-[[2-(trifluoromethyl)phenyl]methoxy]thiophene-2-carboxamide |
| Synonyms | GW843682X; GW-843682X; GW 843682X |
| 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 = 2.2 nM); PLK3 (IC50 = 9.1 nM); PDGFR1β (IC50 = 160 nM); VEGFR2 (IC50 = 360 nM); Aurora A (IC50 = 4800 nM); CDK2/cyclin A (IC50 = 7600 nM) |
| ln Vitro | GW843682X (compound 1) is effectively suppresses the growth of tumor cells, as demonstrated by IC50 values of 0.41, 0.57, 0.11, 0.38, and 0.70 μM for HeLa, A549, H460, and HCT116 cell lines. With an IC50 of 0.14 μM, GW843682X dose-dependently inhibits PLK1 phosphorylation of Ser15-p53. GW843682X (3 μM) treatment for 24, 48, and 72 hours results in a strong G2-M arrest in HDF cells and H460 cells. GW843682X (5 μM) causes H460 cells to undergo apoptosis as opposed to HDF cells[1]. At a 120 nM EC50, GW843682X suppresses the growth of U937 cells. U937 cells' 50% mitotic entry is suppressed when GW843682X (500 nM) and 5 µM VP-16 are combined[2]. Inhibiting the growth of acute leukemia cells, GW843682X (0.06-1μM) enhances vincristine's anti-proliferative qualities. Leukemia cells undergo apoptosis when exposed to GW843682X (0.1–1 μM) in a manner that is dependent on both time and dose. Bcl-xl is dephosphorylated in leukemia cells by GW843682X (0.5–1 μM)[3]. |
| Enzyme Assay | Trichoplusia ni cells infected with baculoviruses are used to prepare PLK1 and PLK3 proteins. The following is the method used to determine PLK1 and PLK3 enzyme activity. Every measurement was made in an environment where the amount of signal produced rose linearly with the enzyme and time. Test compounds are added at varying known concentrations in 100% DMSO to white 384-well assay plates (0.1 μL for 10 μL and some 20 μL assays, 1 μL for some 20 μL assays). As controls, DMSO (1-5% final) and EDTA (65 mM) are employed. The components of the reaction mix at 22°C are as follows: pH 7.2 HEPES with a concentration of 25 mM, 15 mM MgCl2, 1 μM ATP, 0.05 μCi/well [γ-33P]ATP (10 Ci/mmol), 1 μM substrate peptide (Biotin-Ahx-SFNDTLDFD), 0.15 mg/mL bovine serum albumin, 1 mM DTT, and either 2 nM PLK1 kinase domain or 5 nM full-length PLK3. Using automated liquid handlers, quickly add 10 or 20 μL of Reaction Mix to each well after adding the enzyme, and then incubate for 1 to 1.5 hours at 22°C. Using 50 μL of stop mix [50 mM EDTA, 4.0 mg/mL streptavidin SPA beads in Dulbecco's PBS (without Mg2+ and Ca2+), 50 μM ATP] per well, the 20 μL enzymatic reactions are terminated. Ten microliters (50 microliters) of stop mix—three milliliters of streptavidin-coupled SPA Imaging beads in Dulbecco's PBS (without Mg2+ and Ca2+) and fifty microliters of EDTA—per well are used to halt the ten microliter reactions. The plates are either sealed, spun at 500 × g for a minute or left to settle overnight, and then counted in Packard TopCount for 30 seconds per well or imaged using a Viewlux imager. In comparison to the results obtained in control (DMSO-only) wells, the signal above background (EDTA controls) is converted to a percent inhibition[1]. |
| Cell Assay | Data analysis and experimentation are done. Specifically, in these assays, 2,000 H460 cells are plated per well, 5,000 HDF cells are plated per well, and 7,000 and 6,000 per well, respectively, are plated in a 96-well plate for the drug-resistant cell line MES-SA/DX5 and its sensitive parent line MES-SA. Throughout the three days of the experiment, these densities enabled the vehicle controls to grow logarithmically. The compound (30-0.00152 μM) is subjected to threefold dilutions in three different media: low-glucose DMEM, which contains 5% FBS, 50 μg/mL gentamicin, and 0.3% (v/v) DMSO (HDF cells); RPMI 1640, which contains 5% FBS, 50 μg/mL gentamicin, and 0.3% (v/v) DMSO (H460); or McCoy's 5A, which contains 5% FBS, 50 μg/mL gentamicin, and 0.3% (v/v) DMSO (MES-SA and MES-SA/DX5)[1]. |
| References |
[1]. In vitro biological activity of a novel small-molecule inhibitor of polo-like kinase 1. Mol Cancer Ther. 2007 Feb;6(2):450-9. Epub 2007 Jan 31. [2]. Evaluation of Polo-like Kinase 1 inhibition on the G2/M checkpoint in Acute Myelocytic Leukaemia. Eur J Pharmacol. 2008 Sep 4;591(1-3):102-5. [3]. A novel treatment strategy targeting polo-like kinase 1 in hematological malignancies. Leukemia. 2009 Sep;23(9):1564-76. |
Solubility Data
| Solubility (In Vitro) |
DMSO: 33.3~95 mg/mL (69.8~199 mM) Ethanol: ~3 mg/mL (~6.3 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (5.24 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. 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 (5.24 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0944 mL | 10.4721 mL | 20.9442 mL | |
| 5 mM | 0.4189 mL | 2.0944 mL | 4.1888 mL | |
| 10 mM | 0.2094 mL | 1.0472 mL | 2.0944 mL |