NH125

Catalog No.S7436 Batch:S743601

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Technical Data

Formula

C27H45IN2
Molecular Weight 524.56 CAS No. 278603-08-0
Solubility (25°C)* In vitro DMSO 100 mg/mL (190.63 mM)
Ethanol 100 mg/mL (190.63 mM)
Water Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description NH125 is a selective eEF-2 kinase inhibitor with IC50 of 60 nM, >125-fold selectivity over PKC, PKA, and CaMKII, and also a potent histidine kinase inhibitor.
Targets
eEF-2 kinase [1]
60 nM
In vitro 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. [1] 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. [2] EEF2K inhibition by NH125 renders tumor cells more sensitive to curcumin and velcade, which possess ER stress-inducing action. [3]
In vivo NH125 reduces blood pressure in SHR and ROS production, induction of inflammatory molecules, and hypertrophy in SHR superior mesenteric artery. [4]

Protocol (from reference)

Kinase Assay:

[1]
  • eEF-2 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:

[1]
  • Cell lines

    C6, T98-G, U-138 MG, U-87 MG, A172, A2780, HeLa, PC3, OVCAR-3, MCF-7 cell lines.

  • Concentrations

    ~10 μM

  • Incubation Time

    48-72 hours

  • Method

    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.
Animal Study:

[4]
  • Animal Models

    Spontaneously hypertensive rats (SHR)

  • Dosages

    ~500 μg/kg/day

  • Administration

    s.c.

Customer Product Validation

Data from [Data independently produced by , , Acta Pharmacol Sin, 2018, doi：10.1038/s41401-018-0165-9]

Data from [Data independently produced by , , BMC Cancer, 2016, 16(1):813.]

Data from [Data independently produced by , , Drug Des Devel Ther, 2018, 12:2655-2663]

Selleck's NH125 has been cited by 3 publications

Tubeimoside-1, a triterpenoid saponin, induces cytoprotective autophagy in human breast cancer cells in vitro via Akt-mediated pathway [Jiang SL Acta Pharmacol Sin, 2018, 10.1038/s41401-018-0165-9] PubMed: 30315250
Inhibiting eEF-2 kinase-mediated autophagy enhanced the cytocidal effect of AKT inhibitor on human nasopharyngeal carcinoma [Zhao YY Drug Des Devel Ther, 2018, Drug Des Devel Ther] PubMed: 30214154
Inhibition of eEF-2 kinase sensitizes human nasopharyngeal carcinoma cells to lapatinib-induced apoptosis through the Src and Erk pathways [Liu L, et al. BMC Cancer, 2016, 16(1):813] PubMed: 27756261

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.