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NH125 CaMK inhibitor

Cat.No.S7436

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.
NH125 CaMK inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 524.56

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Quality Control

Batch: S743601 DMSO]100 mg/mL]false]Ethanol]100 mg/mL]false]Water]Insoluble]false Purity: 99.91%
99.91

Chemical Information, Storage & Stability

Molecular Weight 524.56 Formula

C27H45IN2

 Storage (From the date of receipt)
CAS No. 278603-08-0 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles CCCCCCCCCCCCCCCCN1C=C[N+](=C1C)CC2=CC=CC=C2.[I-]

Solubility

In vitro
Batch:

DMSO : 100 mg/mL (190.63 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 100 mg/mL

Water : Insoluble

Molarity Calculator

Mass Concentration Volume Molecular Weight

In vivo
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Mechanism of Action

Targets/IC50/Ki
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. This compound potently inhibits cell viability of 10 cancer cells with IC50 ranging from 0.7 to 4.8 μM. [1] It 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 this chemical renders tumor cells more sensitive to curcumin and velcade, which possess ER stress-inducing action. [3]
Kinase Assay
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.
In vivo
NH125 reduces blood pressure in SHR and ROS production, induction of inflammatory molecules, and hypertrophy in SHR superior mesenteric artery. [4]
References

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