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Purity: ≥98%
GSK2606414 is a novel, potent and selective inhibitor of PERK (PRKR-like endoplasmic reticulum kinase or protein kinase R (PKR)-like endoplasmic reticulum kinase) with an IC50 of 0.4 nM and at least 100-fold selectivity over the other EIF2AKs tested. Another name for PERK is EIF2AK3 (Eukaryotic translation initiation factor 2-alpha kinase 3). The EIF2AK3 gene encodes PERK. It is a member of the family of membrane protein type I. It occurs in the endoplasmic reticulum (ER) and is brought on by ER stress, which is brought on by improperly folded proteins.
Targets |
EIF2AK3 (PERK) (IC50 = 0.4 nM); EIF2AK1 (HRI) (IC50 = 420 nM); EIF2AK2 (PKR) (IC50 = 696 nM)
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ln Vitro |
GSK2606414 inhibits PERK activation in cells[1].
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ln Vivo |
GSK2606414 shows low to moderate blood clearance and high oral availability in mouse, rat, and dog. Oral administration of GSK2606414 inhibits tumor growth in mice with pancreatic human BxPC3 tumors in a dose-dependent manner. [1]
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Enzyme Assay |
The cytoplasmic domain of GST-PERK is bought. 6-Human eIF2α full-length is isolated from baculovirus expression in Sf9 insect cells. The eIF2α protein is buffer exchanged by dialysis into PBS, followed by chemical modification with NHS-LC-biotin and buffer exchanged into 50 mM Tris, pH 7.2, 250 mM NaCl, and 5 mM DTT. Aliquoted protein is kept at -80°C for storage. When the freshly prepared quench solution is added to the reaction, the final concentrations of 4 nM eIF2α phospho-ser51-antibody, 4 nM anti-rabbit IgG labeled with Eu-1024, 40 nM streptavidin Surelight APC, and 15 mM EDTA are obtained. At a final volume of 10 μL, reactions were carried out in black 384-well polystyrene low volume plates. 10 mM HEPES, 5 mM MgCl2, 5 μM ATP, 1 mM DTT, 2 mM CHAPS, 40 nM biotinylated 6-His-eIF2α, and 0.4 nM GST-PERK are all present in the reaction volume in final concentrations. The compounds being analyzed are dissolved in 1.0 mM DMSO and then serially diluted 1–3 with DMSO over the course of 11 dilutions. For every concentration, 0.1 μL is added to the appropriate well in an assay plate. This results in a final compound concentration range of 10 μM to 0.00017 μM. Assay plates containing compounds are preincubated for 30 minutes at room temperature before receiving the GST-PERK solution. The addition of ATP and the eIF2α substrate solution starts the reaction. The quench solution is added after an hour of incubation. Before the signal is determined, the plates are covered and left at room temperature for two hours. Using a Viewlux reader, the resulting signal is quantified. An APC/Eu calculation is used to transform the data and normalize the APC signal to the europium signal. The data reduction formula 100 × [1 – (U1 – C2)/(C1 – C2)] is used to plot the percentage inhibition against the compound concentration for concentration response curves. In this formula, U represents the unknown value, C1 represents the average control value obtained for 1% DMSO, and C2 represents the average control value obtained for 0.1 M EDTA. A, B, C, D, and x represent the compound concentration, IC50, slope factor, and minimum and maximum responses, respectively, on a curve that fits the data. As IC50 values, the outcomes are noted for every compound.
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Cell Assay |
Thapsigargin was used to stimulate the cells after PERK inhibitor (GSK2606414, compound 38) incubation. The cells were lysed and examined for PERK autophosphorylation inhibition after two hours.
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Animal Protocol |
Female nude mice have their right flank subcutaneously implanted with exponentially growing BxPC3 tumor cells (10×106 cells/mouse) cultivated in cell culture. Mice with tumors measuring less than 200 mm3 are randomly assigned to one of eight treatment groups sixteen days post-implantation. The animals are given the compound at 50 or 150 mg/kg, b.i.d., for 21 days along with a vehicle (0.5% hydoxypropylmethylcellulose, 0.1% Tween 80 in water, pH 4.8). Using calipers, the tumor volume is measured twice a week and computed. When the dosage is finished, the results are expressed as a percent inhibition, or 100[1-(average growth of the drug-treated population)/(average growth of the vehicle-treated control population)]. In statistical analysis, the two-tailed t test is employed.
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References |
Molecular Formula |
C24H20F3N5O
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Molecular Weight |
451.44
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Exact Mass |
451.16199
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Elemental Analysis |
C, 63.85; H, 4.47; F, 12.63; N, 15.51; O, 3.54
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CAS # |
1337531-36-8
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Appearance |
white solid powder
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SMILES |
CN1C=C(C2=C(N=CN=C21)N)C3=CC4=C(C=C3)N(CC4)C(=O)CC5=CC(=CC=C5)C(F)(F)F
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InChi Key |
SIXVRXARNAVBTC-UHFFFAOYSA-N
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InChi Code |
InChI=1S/C24H20F3N5O/c1-31-12-18(21-22(28)29-13-30-23(21)31)15-5-6-19-16(11-15)7-8-32(19)20(33)10-14-3-2-4-17(9-14)24(25,26)27/h2-6,9,11-13H,7-8,10H2,1H3,(H2,28,29,30)
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Chemical Name |
1-[5-(4-amino-7-methylpyrrolo[2,3-d]pyrimidin-5-yl)-2,3-dihydroindol-1-yl]-2-[3-(trifluoromethyl)phenyl]ethanone
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Synonyms |
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HS Tariff Code |
2934.99.9001
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Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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Solubility (In Vitro) |
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Solubility (In Vivo) |
5% DMSO+45% PEG 300+ddH2O: 20mg/mL (Please use freshly prepared in vivo formulations for optimal results.)
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Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 2.2151 mL | 11.0757 mL | 22.1513 mL | |
5 mM | 0.4430 mL | 2.2151 mL | 4.4303 mL | |
10 mM | 0.2215 mL | 1.1076 mL | 2.2151 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.
Calculation results
Working concentration: mg/mL;
Method for preparing DMSO stock solution: mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation::Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
PK4 inhibitor GSK2606414 prevents recrudescence following ART treatment.Cell Host Microbe. 2017 Dec 13;22(6):766-776.e4. th> |
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Phosphorylation of PfeIF2α in aPfK13C580Y mutant parasite.Cell Host Microbe. 2017 Dec 13;22(6):766-776.e4. td> |