RIPK2 Inhibitor 1
Catalog No.: 19510-BID002
RIPK2 (Receptor Interacting Protein Kinase 2) is the obligate kinase of the NOD2 intracellular pathogen recognition pathway receptor and an upstream kinase activator for NFκB via activation of TAK1. Inhibiting the activity of RIPK2 may be important in controlling inflammatory responses.
Alternative Name: 3-benzamido-4-methyl-N-[3-(1-methyl-1H-imidazol-2yl)phenyl] benzamide.
Molecular Weight: 410.47g/mol
Solubility: Stocks should be made up in 100% DMSO to 10-20mM. Stocks can be diluted in >12% DMSO to desired concentration or dissolved in 100% olive oil.
In vitro: IC50 is 5-50 nM depending on cell type.
Cell-based inhibition kinetics
Left side: Inhibition was carried out for 24-36 hours on Hodgkins lymphoma cell line KMH2 revealing constitutively active RIPK2. This was followed by lysis and immunoprecipitation (IP) with rabbit anti-RIPK2 antibody (#19510) overnight for total RIPK2 pull down. Following Protein G pull down and wash, 32P-γ-ATP is added and kinase reaction allowed to proceed for 30 minutes at 30°C. Following kinase reaction, SDS-PAGE was used to separate out the proteins, gel was then dried and exposed to film. The p60 RIPK2 band was quantitated.
Right side: NFκB gene reporter assay determination of inhibition of MDP stimulated NFκB activity with #19510-BID002. An approximate EC50 determination for inhibition of MDP driven-NFκB activation is shown.
RIPK2 Inhibitor 1
In vivo: May be evaluated at 1-15ug/g body weight.
In vivo inhibition of DSS-induced colitis
1a-/- of 1a+/- Wild type Percent Survival Days after 3% DSS addition 1a -/- + RIPK2 Inhibitor 1 (D3, 5, 7) Add 3% DSS Add WATER 0 5 10 15 20 0 20 40 60 80 100 1a +/- + RIPK2 Inhibitor 1 (D3, 5, 7)
DSS-induced inflammation can be inhibited by RIPK2 inhibitor 1 at 1ug/g body weight injected intraperitoneally (IP) at day 3, 5 and 7 (during period of active disease and time of recovery). The Kaplan- Meir survival curve is plotted for Rassf 1a-/- and 1a+/- mice. Both potentially have RIPK2 unchecked in colonic tissue (and hence elevated inflammation) if Rassf1a is missing. N = 12-20 animals per condition;
p value > 0.001.
STORAGE AND STABILITY
Store at -80°C in appropriate aliquots to avoid multiple freeze-thaw cycles.
Salla M et al. Identification and Characterization of Novel Receptor-Interacting Serine/Threonine- Protein Kinase 2 Inhibitors using structural similarity analysis. 2018 J Pharmacol Exp Ther 365: 354-367.
For investigational use only. Not for use in therapeutic or diagnostic procedures.