GSK2334470 (GSK-2334470) is a novel, highly specific and potent inhibitor of PDK1 (3-phosphoinositide-dependent protein kinase) with potential antitumor activity.In a cell-free assay, it inhibits PDK-1 with an IC50 of 10 nM and has no impact on closely related AGC-kinases. GSK2334470 inhibited PDK1 from activating full-length Akt1 or mutant Akt1 lacking the PH domain, which resulted in strong antiproliferative activity in vitro and high antitumor efficacy in vivo.
Physicochemical Properties
| Molecular Formula | C25H34N8O | |
| Molecular Weight | 462.5905 | |
| Exact Mass | 462.285 | |
| Elemental Analysis | C, 64.91; H, 7.41; N, 24.22; O, 3.46 | |
| CAS # | 1227911-45-6 | |
| Related CAS # |
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| PubChem CID | 46215815 | |
| Appearance | white solid powder | |
| Density | 1.3±0.1 g/cm3 | |
| Index of Refraction | 1.667 | |
| LogP | 2.31 | |
| Hydrogen Bond Donor Count | 4 | |
| Hydrogen Bond Acceptor Count | 7 | |
| Rotatable Bond Count | 5 | |
| Heavy Atom Count | 34 | |
| Complexity | 688 | |
| Defined Atom Stereocenter Count | 2 | |
| SMILES | O=C([C@]1([H])C([H])([H])N(C2C([H])=C(C3C([H])=C([H])C4C(N([H])[H])=NN([H])C=4C=3[H])N=C(N([H])C([H])([H])[H])N=2)[C@]([H])(C([H])([H])[H])C([H])([H])C1([H])[H])N([H])C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C1([H])[H] |
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| InChi Key | QLPHOXTXAKOFMU-WBVHZDCISA-N | |
| InChi Code | InChI=1S/C25H34N8O/c1-15-8-9-17(24(34)28-18-6-4-3-5-7-18)14-33(15)22-13-20(29-25(27-2)30-22)16-10-11-19-21(12-16)31-32-23(19)26/h10-13,15,17-18H,3-9,14H2,1-2H3,(H,28,34)(H3,26,31,32)(H,27,29,30)/t15-,17+/m1/s1 | |
| Chemical Name | (3S,6R)-1-(6-(3-amino-1H-indazol-6-yl)-2-(methylamino)pyrimidin-4-yl)-N-cyclohexyl-6-methylpiperidine-3-carboxamide | |
| Synonyms | GSK 2334470; GSK2334470; GSK-2334470 | |
| 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 |
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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) |
Biological Activity
| Targets | PDK1 (IC50 = 10 nM) |
| ln Vitro | GSK2334470 inhibits PDK1 from activating full-length Akt1 in the presence of PtdIns(3,4,5)P3-containing lipid vesicles or a mutant of Akt1 lacking the PH domain (ΔPH-Akt1) with IC50 of ~10 nM. With an IC50 of less than 10 nM, GSK2334470 similarly prevents PDK1 from phosphorylating the PDKtide peptide substrate. In HEK-293 cells, GSK2334470 (0.1 M-0.3 M) causes a significant dose-dependent inhibition of endogenous NDRG1 with a phosphorylation reduction of over 50%. In HEK-293 cells, GSK2334470 (30 nM) causes a significant, dose-dependent inhibition of each SGK isoform's T-loop phosphorylation. GSK2334470 (30 µM) suppresses U46619 induced Ca2+-sensitized force in α-toxin permeabilized rabbit pulmonary artery SM. GSK2334470 (30 µM) results in a significant decrease in the contractile force in response to [Ca2+]. [2]GSK2334470 (1 μM) results in total abrogation of the EGF-induced intracellular calcium increase and inositol phosphates accumulation in MDA-MB-231 cells. GSK2334470 (1 μM) inhibits PLCγ1 Tyr783 phosphorylation in MDA-MB-231 cells. [3] |
| ln Vivo | GSK2334470 is a highly specific and potent PDK1 inhibitor. |
| Enzyme Assay | Endogenous Akt, S6K, and RSK are immunoprecipitated using 3 g to 5 g of the designated antibodies from 0.1 mg to 1 mg of cell lysate for two hours at 4 °C on a vibrating platform. In order to measure the SGK activity, 150 g of transfected lysate is incubated with 5 g of glutathione-Sepharose at 4 °C for 3 hours. The immunoprecipitates are then washed twice with kinase buffer and twice with lysis buffer containing 0.5 mM NaCl. A reaction mixture is used to start kinase reactions, resulting in final concentrations of 0.1 mM [-32P]ATP (200 c.p.m./pmol), 5 mM magnesium acetate, 0.1% 2-mercaptoethanol, and 30 mM Crosstide peptide (GRPRTSSFAEGKK). |
| Cell Assay | GSK2334470 is dissolved in DMSO and diluted with appropriate medium before use. GSK2334470 is administered to non-resistant cells and resistant sublines for 1.5 and 12 hours in 10% FBS medium with/without MK-2206 (5 M) to investigate its inhibitory effect on the mTOR-S6K pathway. |
| Animal Protocol | In order to dilute the mice, DMSO is first dissolved in PBS or saline. As previously mentioned, BrafV600E::Pten are generated. Each experimental group utilizes cohorts of six animals. Based on previous studies[3], GSK2334470 is administered by IP injection (100 mg/kg) three times per week beginning on the day that 4-hydroxytamoxifen is topically applied and ending at the time of mouse collection. |
| References |
[1]. Biochem J. 2011 Jan 15;433(2):357-69. [2]. Cancer Cell Int. 2015 Sep 29;15:91. [3]. J Cell Sci. 2012 Jul 1;125(Pt 13):3153-63. |
| Additional Infomation | (3S,6R)-1-[6-(3-amino-1H-indazol-6-yl)-2-(methylamino)-4-pyrimidinyl]-N-cyclohexyl-6-methyl-3-piperidinecarboxamide is a member of indazoles. |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~90 mg/mL (194.6 mM) Water: <1 mg/mL Ethanol: 90 mg/mL (194.6 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.40 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 (5.40 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 (5.40 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 | 2.1617 mL | 10.8087 mL | 21.6174 mL | |
| 5 mM | 0.4323 mL | 2.1617 mL | 4.3235 mL | |
| 10 mM | 0.2162 mL | 1.0809 mL | 2.1617 mL |