Bardoxolone Methyl

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Biological Activity

Description	Bardoxolone Methyl is an IKK inhibitor, showing potent proapoptotic and anti-inflammatory activities. Phase 3.
Targets	IKK
IC50	[3]
In vitro	Bardoxolone Methyl exhibits potent inhibitory activities against production of nitric oxide induced by interferon-Ƴ in mouse macrophages with IC50 of 0.1 nM. [1] Bardoxolone Methyl decreases the viability of leukemic HL-60, KG-1, and NB4 cells with IC50 of 0.4, 0.4, and 0.27 μM, respectively. CDDO-Me induces pro-apoptotic Bax protein, inhibits the activation of ERK1/2, and it blocks Bcl-2 phosphorylation, which contributes to the induction of apoptosis. [2] Bardoxolone Methyl potently inhibits both constitutive and inducible NF-kappaB activated by TNF, interleukin (IL)-1beta, phorbol ester, okadaic acid, hydrogen peroxide, lipopolysaccharide, and cigarette smoke. [3]
In vivo	Bardoxolone Methyl (60 mg/kg) reduces the number, size, and severity of lung tumors in vivo. [4] Bardoxolone Methyl significantly reduces the in vivo inflammatory cytokine response following LPS challenge, induces HO-1 protein expression in the spleen, and protects mice against lethal-dose LPS. [5]
Features	The only IKKβ inhibitor in clinical use for solid tumors, type 2 diabetes, and chronic kidney disease. An orally-available antioxidant inflammation modulator.

Protocol(Only for Reference)

Kinase Assay: [3]

IKK assay

To determine the effect of CDDO-Me on TNF-induced IKK activation, IKK is analyzed. Briefly, the IKK complex from whole-cell extracts was precipitated with antibody against IKKα and IKKβ and then treated with protein A/G-Sepharose beads. After 2 hours, the beads are washed with lysis buffer and then resuspended in a kinase assay mixture containing 50 mmol/L HEPES (pH 7.4), 20 mmol/L MgCl2, 2 mmol/L DTT, 20 μCi [γ-32P]ATP, 10 μmol/L unlabeled ATP, and 2 μg of substrate glutathione S-transferase-IκBα (amino acids 1-54). After incubation at 30°C for 30 minutes, the reaction is terminated by boiling with SDS sample buffer for 5 minutes. Finally, the protein is resolved on 10% SDS-PAGE, the gel is dried, and the radioactive bands are visualized with a Storm820. To determine the total amounts of IKK-α and IKK-β in each sample, 50 μg of whole-cell proteins are resolved on 7.5% SDS-PAGE, electrotransferred to a nitrocellulose membrane, and then blotted with either anti-IKK-α or anti-IKK-β antibody.

Cell Assay: [2]

Cell lines	HL-60, KG-1, and NB4 cells
Concentrations	~5 μM
Incubation Time	72 hours
Method	Leukemic cell lines are cultured at a density of 3.0 × 105 cells/mL, and AML mononuclear cells are cultured at 5 × 105 cells/mL in the presence or absence of indicated concentrations of CDDO-Me. Appropriate amounts of DMSO (final concentration less than 0.05%) are included as control. For cytotoxicity studies, 1 μM ara-C is added to the cultures. After 24 to 72 hours, viable cells are counted with the trypan blue dye exclusion method using a hematocytometer.

Animal Study: [4]

Animal Models	Female A/J mice are injected i.p. with vinyl carbamate.
Dosages	~60 mg/kg
Administration	Oral gavage
Solubility	1% DMSO/30% polyethylene glycol/1% Tween 80, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01689116	Completed	Healthy Volunteers	Reata Pharmaceuticals, Inc.	2012-08	Phase 1
NCT01563562	Completed	Hepatic Impairment|Healthy	Reata Pharmaceuticals, Inc.	2012-08	Phase 1
NCT01549769	Terminated	Renal Insufficiency, Chronic|Diabetes Mellitus, Type 2	Reata Pharmaceuticals, Inc.	2012-08	Phase 1
NCT01551446	Withdrawn	Renal Insufficiency, Chronic|Diabetes Mellitus, Type 2	Reata Pharmaceuticals, Inc.	2012-08	Phase 1
NCT01655186	Withdrawn	Renal Insufficiency, Chronic|Diabetes Mellitus, Type 2	Reata Pharmaceuticals, Inc.	2012-09	Phase 2

Chemical Information

Download Bardoxolone Methyl SDF

Chemical Name	Oleana-1,9(11)-dien-28-oic acid, 2-cyano-3,12-dioxo-, methyl ester