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    InvivoChem Cat #: V1297
    CAS #: 278603-08-0Purity ≥98%

    Description: NH125 (NH-125; NH 125) is a potent and selective eEF-2 (eukaryotic elongation factor 2) kinase inhibitor with antibacterial activity against various Gram-positive and -negative bacteria. It inhibits eEF-2 with an IC50 of 60 nM, and showed >125-fold selectivity over PKC, PKA, and CaMKI. NH125 is also a potent histidine kinase inhibitor. When tested with a panel of 10 cancer cell lines (C6, T98-G, U-138 MG, and so forth), NH125 treatment inhibited cell viability with IC50 value ranges from 0.7 to 4.8 μM. NH125 decreased the cellular content of p-eEF-2 without affecting total content eEF-2 and arrested cell in G0-G1 phase in C6 glioma cells. When tested with a panel of human cancer cell lines (glioblastoma, breast cancer, and so on), NH125 sensitized cells at the dose of 0.25 μM which thus reinforced the efficacy of ER stress-inducing drug by inhibiting eEF-2.    

    References: Cancer Res. 2003 Oct 15;63(20):6894-9; Am J Physiol Heart Circ Physiol. 2013 Sep 1;305(5):H756-68.

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    Molecular Weight (MW)524.56
    CAS No.278603-08-0
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 100 mg/mL (190.6 mM)
    Water:<1 mg/mL
    Ethanol: 100 mg/mL (190.6 mM)
    Other info

    Chemical Name: 1-Hexadecyl-2-methyl-3-(phenylmethyl)-1H-imidazolium iodide


    InChi Code: InChI=1S/C27H45N2.HI/c1-3-4-5-6-7-8-9-10-11-12-13-14-15-19-22-28-23-24-29(26(28)2)25-27-20-17-16-18-21-27;/h16-18,20-21,23-24H,3-15,19,22,25H2,1-2H3;1H/q+1;/p-1



    NH-125; NH 125; NH125

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    In Vitro

    In vitro activity: In C6 glioma cells, NH125 decreases the cellular content of phospho-eEF-2 without affecting total content eEF-2 content, and blocks cell cycle transit at the G1-S boundary. NH125 potently inhibits cell viability of 10 cancer cells with IC50 ranging from 0.7 to 4.8 μM. NH125 effectively inhibits histidine protein kinases, including Envz, PhoQ, BvgS, EvgS, and thus produces potent anti-bacteria activities on oxacillin-resistant Staphylococcus aureus (ORSA), vancomycin-resistant Enterococcus faecalis (VRE), penicillin-resistant Streptococcus pneumoniae (PRS), and other Gram-positive and Gram-negative bacteria. EEF2K inhibition by NH125 renders tumor cells more sensitive to curcumin and velcade, which possess ER stress-inducing action.

    Kinase Assay: eEF-2 kinase activity is measured by two methods: (a) a filter-based assay; and (b) by immunoblotting using antiphospho-eEF2 antibody. For both of these, reactions are carried out in 20 μl of total volume containing 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1.5 mM CaCl2, 100 μg/ml calmodulin, 2 μM His-tagged eEF-2 and 400 nM GST-eEF-2 kinase, and ATP mixture [50 μM ATP with 1μCi (γ-33P)ATP]. The kinase mixture without ATP is prepared on ice and then preincubated for 15 min at room temperature. Kinase reactions are started by adding ATP and allowed to progress at 30°C for 30 min. For the filter-based assay, the reaction is terminated by adding 20 μl of cold 1.5% phosphoric acid, and 5 μl of the reaction are applied to P81 Whatman phosphocellulose paper. The paper is washed three times in 500 ml of 0.5% phosphoric acid and once with 200 ml of acetone. The paper is then air-dried and immersed in 10 ml of scintillation mixture. Radioactivity is counted using a Beckton-Dickinson liquid scintillation counter. For immunoblotting, the reactions are stopped by addition of 20 μl of 3× Lamelli buffer [190 mM Tris (pH 6.8), 6% SDS, 30% glycerol, 15% 2-mercaptoethanol, and 0.003% bromphenol blue dye]. Samples are boiled for 5 min and resolved by 7% SDS-PAGE and processed for Western blotting as described below. Conditions for both assays are chosen to ensure linearity of the reaction with respect to time of incubation and concentration of enzyme.

    Cell Assay: The viability of cells is measured using an MTT assay. Briefly, 5 × 104 cells are plated in 96-well plates and exposed to various concentrations of drug for 48–72 h. The formazan product formed after 4 h incubation with MTT is dissolved in 100% DMSO and read at 550 nM using a Dynatech Microplate Reader MR5000.

    In VivoNH125 reduces blood pressure in SHR and ROS production, induction of inflammatory molecules, and hypertrophy in SHR superior mesenteric artery.
    Animal modelSpontaneously hypertensive rats (SHR)
    Formulation & DosageDissolved in DMSO; ~500 μg/kg/day; s.c.

    Cancer Res. 2003 Oct 15;63(20):6894-9; Am J Physiol Heart Circ Physiol. 2013 Sep 1;305(5):H756-68.

    These protocols are for reference only. InvivoChem does not independently validate these methods.


    Effects of eukaryotic elongation factor (eEF) 2 kinase (eEF2K) inhibitor NH125 or A-484954 on TNF-α-induced expression of VCAM-1 (A, n = 5; C, n = 6–8) and endothelial-selectin (E-selectin; B, n = 6; D, n = 4) in cultured human umbilical vein endothelial cells (ECs) (HUVECs). Am J Physiol Heart Circ Physiol. 2013 Sep 1;305(5):H756-68.


    Effects of NH125 on TNF-α-induced phosphorylation of JNK (A) and NF-κB (B), and ROS production (C) in HUVECs. Am J Physiol Heart Circ Physiol. 2013 Sep 1;305(5):H756-68.


    Effects of long-term NH125 treatment on expression of eEF2K (A), VCAM-1 (B), and E-selectin (C), and ROS production (D) in isolated mesenteric artery. Am J Physiol Heart Circ Physiol. 2013 Sep 1;305(5):H756-68. 


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