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Bardoxolone Methyl (RTA 402) NRF2 Activator

Name: Bardoxolone Methyl (RTA 402)
Brand: Selleck Chemicals
SKU: S8078
Availability: InStock
Rating: 5 (36 reviews)

Cat.No.S8078

Bardoxolone Methyl (RTA 402, TP-155, NSC 713200, CDDO Methyl Ester, CDDO-Me) is an IKK inhibitor, showing potent proapoptotic and anti-inflammatory activities; Also a potent Nrf2 activator and nuclear factor-κB (NF-κB) inhibitor. Bardoxolone Methyl abrogates ferroptosis. Bardoxolone methyl induces apoptosis and autophagy in cancer cells.

Chemical Structure

Molecular Weight: 505.69

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Quality Control

Batch: Purity: 99.86%

99.86

Related Targets	NF-κB HDAC Antioxidant ROS IκB/IKK AP-1 MALT NOD
Other Nrf2 Inhibitors	ML385 Brusatol Sulforaphane TBHQ Oltipraz Bardoxolone (CDDO) 4-Octyl Itaconate (4-OI) KI696 CDDO-Im (RTA-403) cis-Resveratrol

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
MCF-7	Function assay			Inhibitory concentration against proliferation of MCF-7 (ER Positive) breast cancer cells, IC50=0.05μM	15369396
BMDM	Cytotoxicity assay		24 hrs	Cytotoxicity against C57BL/6 mouse BMDM cells assessed as LDH release after 24 hrs, MNTD=0.5μM	22533790
BMDM	Antiinflammatory assay	0.5 uM	1 hr	Antiinflammatory activity in C57BL/6 mouse BMDM cells assessed as inhibition of LPS-stimulated TNFalpha production at 0.5 uM pretreated for 1 hr before LPS challenge after 8 to 24 hrs by immunoassay	22533790
PANC1343	Antiproliferative assay	300 to 1000 nM	72 hrs	Antiproliferative activity against mouse PANC1343 cells at 300 to 1000 nM after 72 hrs by MTT assay	24388806
RAW264.7	Antioxidant assay	100 nM	18 hrs	Antioxidant activity in mouse RAW264.7 cells assessed as inhibition of tBHP-induced ROS production at 100 nM pretreated for 18 hrs before challenge measured after 15 mins by H2DCFA-based flow cytometry	24388806
HepG2	Cytotoxicity assay		48 hrs	Cytotoxicity against human HepG2 cells after 48 hrs by MTT assay, IC50=4.99μM	24685545
B16F10	Cytotoxicity assay		48 hrs	Cytotoxicity against mouse B16F10 cells after 48 hrs by MTT assay, IC50=5.85μM	24685545
CCD-841-CoN	Antiproliferative assay		72 hrs	Antiproliferative activity against human CCD-841-CoN cells assessed as inhibition of cell proliferation after 72 hrs by MTT assay, IC50=0.316μM	25675144
HCT8	Antiproliferative assay		72 hrs	Antiproliferative activity against 5-FU resistant human HCT8 cells assessed as inhibition of cell proliferation after 72 hrs by MTT assay, IC50=0.363μM	25675144
HCT8	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT8 cells assessed as inhibition of cell proliferation after 72 hrs by MTT assay, IC50=0.399μM	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of HIF-1alpha protein expression in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of HIF-1alpha protein expression in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of STAT3 protein phosphorylation in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of STAT3 protein phosphorylation in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of AKT protein phosphorylation in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of AKT protein phosphorylation in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of ERK protein phosphorylation in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of ERK protein phosphorylation in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Antiproliferative assay	1 uM	72 hrs	Antiproliferative activity against human HCT8 cells assessed as inhibition of cell proliferation at 1 uM after 72 hrs by MTT assay	25675144
HCT8	Antiproliferative assay	1 uM	72 hrs	Antiproliferative activity against 5-FU resistant human HCT8 cells assessed as inhibition of cell proliferation at 1 uM after 72 hrs by MTT assay	25675144
BEAS2B	Function assay	10 uM	6 hrs	Activation of Nrf2 in human BEAS2B cells assessed as increase in HO1 gene expression at 10 uM incubated for 6 hrs by qPCR method	26278028
H42E	Function assay		24 hrs	Induction of NRF2 activation in rat H42E cells expressing ARE8L assessed as reporter transgene activity after 24 hrs by luminescence assay, CD=0.0005μM	26908173
H42E	Cytotoxicity assay		24 hrs	Cytotoxicity against rat H42E cells expressing ARE8L assessed as cellular ATP level after 24 hrs by Celltiter-Glo luminescent cell viability assay, IC50=1.4μM	26908173
H42E	Function assay	0.01 to 30 nM	1 hr	Stabilization of NRF2 in rat H42E cells expressing ARE8L at 0.01 to 30 nM after 1 hr by Western blot analysis	26908173
NHBE	Cytoprotective assay	0.001 to 0.1 uM	18 hrs	Cytoprotective activity in NHBE cells assessed as inhibition of tBHP-induced GSH depletion at 0.001 to 0.1 uM preincubated for 18 hrs followed by tBHP addition for 4 hrs by thiostar dye based fluorescence assay	27031670
NHBE	Function assay	100 nM	24 hrs	Inhibition of KEAP1/NRF2 interaction in NHBE cells assessed as increase in GCLM mRNA expression at 100 nM incubated for 24 hrs in presence of non targeting siRNA by qRT-PCR method	27031670
NHBE	Function assay	100 nM	24 hrs	Inhibition of KEAP1/NRF2 interaction in NHBE cells assessed as increase in NQO1 mRNA expression at 100 nM incubated for 24 hrs in presence of non targeting siRNA by qRT-PCR method	27031670
NHBE	Function assay	100 nM	48 hrs	Inhibition of KEAP1/NRF2 interaction in NHBE cells assessed as induction of NQO1 specific activity at 100 nM incubated for 48 hrs in presence of non targeting siRNA by MTT reduction assay	27031670
HaCaT-ARE-luc	Function assay		6 hrs	Activation of Nrf2 (unknown origin) expressed in human HaCaT-ARE-luc cells after 6 hrs by luciferase reporter gene assay, EC50=0.06μM	28753294
NIH/3T3	Function assay		6 hrs	Inhibition of TNF-alpha stimulated NF-kappaB (unknown origin) expressed in mouse NIH/3T3 cells after 6 hrs by luciferase reporter gene assay, IC50=1.2μM	28753294
HeLa	Function assay		6 hrs	Inhibition of IFN-gamma stimulated STAT3 (unknown origin) expressed in human HeLa cells after 6 hrs by luciferase reporter gene assay, IC50=2.38μM	28753294
RAW264.7	Anti-inflammatory assay			Anti-inflammatory activity in mouse RAW264.7 cells assessed as inhibition of nitric oxide production, IC50=4μM	28754470
HEK293	Cytotoxicity assay		24 hrs	Cytotoxicity against HEK293 cells assessed as reduction in cell viability after 24 hrs by MTT assay, IC50=2.2μM	28994286
H9c2	Cytotoxicity assay		24 hrs	Cytotoxicity against rat H9c2 cells assessed as reduction in cell viability after 24 hrs by MTT assay, IC50=5.2μM	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as TNFalpha-induced NFkappaB activation at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by NFkappaB-driven luciferase reporter gene assay	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of iNOS mRNA expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by quantitative RT-PCR an	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of COX2 mRNA expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by quantitative RT-PCR an	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of MCP1 mRNA expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by quantitative RT-PCR an	28994286
intraglomerular mesangial cells	Function assay	0.65 mg/kg	12 weeks	Renoprotective activity in db/db mouse assessed as increase in number of intraglomerular mesangial cells at 0.65 mg/kg, ip administered trice per week for 12 consecutive weeks measured at 11 weeks post dose by H/E-staining based microscopic analysis	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of COX2 protein expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot method	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of iNOS protein expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot method	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as mitigation of TNFalpha-induced increase in ratio of nuclear to cytosolic p65 at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of MCP1 protein expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot method	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as upregulation of HO-1 mRNA expression at 200 to 1000 nM after 48 hrs by quantitative RT-PCR analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as upregulation of NQO1 mRNA expression at 200 to 1000 nM after 48 hrs by quantitative RT-PCR analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as activation of Nrf2 at 200 to 1000 nM after 48 hrs by ARE-driven luciferase reporter gene assay	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as increase in nuclear to cytosolic Nfr2 ratio at 200 to 1000 nM after 48 hrs by Western blot analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as increase in cytosolic HO-1 levels at 200 to 1000 nM after 48 hrs by Western blot analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as increase in cytosolic NQO1 levels at 200 to 1000 nM after 48 hrs by Western blot analysis	28994286
A549/TR	Antiproliferative assay		72 hrs	Antiproliferative activity against human A549/TR cells after 72 hrs by MTT assay, IC50=1.703μM	29501947
A549	Antiproliferative assay		72 hrs	Antiproliferative activity against human A549 cells after 72 hrs by MTT assay, IC50=2.074μM	29501947
A549/TR	Function assay	2.4 to 9.6 uM	24 hrs	Induction of ROS generation in human A549/TR cells at 2.4 to 9.6 uM after 24 hrs by DCFH-DA dye-based flow cytometric analysis	29501947
A549/TR	Function assay	4.8 uM	24 hrs	Downregulation of Lon expression in human A549/TR cells at 4.8 uM after 24 hrs by Western blot analysis	29501947
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells after 72 hrs by SRB assay, IC50=0.00025μM	30429953
HT-29	Antiproliferative assay		72 hrs	Antiproliferative activity against human HT-29 cells after 72 hrs by SRB assay, IC50=0.28μM	30429953
HCT8	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT8 cells after 72 hrs by SRB assay, IC50=0.29μM	30429953
HCT116	Function assay		8 hrs	Inhibition of Bmi1 protein expression in human HCT116 cells after 8 hrs by Western blot analysis	30429953
BEAS2B	Function assay		48 hrs	Activation of Keap1/Cul3/Nrf2 in human BEAS2B cells assessed as increase in NQO1 levels measured after 48 hrs, EC50=0.00871μM	30626555
HepG2	Cytotoxicity assay		48 hrs	Cytotoxicity against human HepG2 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay, IC50=0.26μM	31051401
MCF7	Cytotoxicity assay		48 hrs	Cytotoxicity against human MCF7 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay, IC50=0.35μM	31051401
A549	Cytotoxicity assay		48 hrs	Cytotoxicity against human A549 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay, IC50=0.36μM	31051401
A549	Antiproliferative assay		48 hrs	Antiproliferative activity against human A549 cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.52μM	31725288
HepG2	Antiproliferative assay		48 hrs	Antiproliferative activity against human HepG2 cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.52μM	31725288
HOS	Antiproliferative assay		48 hrs	Antiproliferative activity against human HOS cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.66μM	31725288
MCF7	Antiproliferative assay		48 hrs	Antiproliferative activity against human MCF7 cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.85μM	31725288
HEK293FT	Function assay		24 hrs	Inhibition of mouse GOAT expressed in HEK293FT cells co-expressing pre-proghrelin assessed as reduction in ghrelin octanoylation incubated for 24 hrs by ELISA, IC50=0.035μM	ChEMBL
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 26 mg/mL (51.41 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	4%DMSO 1 30%PEG300 2 5%Tween80 3 61%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	2.500mg/ml (4.94mM)	Taking the 1 mL working solution as an example, add 40 μL of clarified DMSO stock solution of 62.5 mg/ml to 300 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify it; then continue to add 610 μL of ddH2O to make it clear. Volume up to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

In vivo Formulation Calculator (Clear solution)

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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

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Chemical Information, Storage & Stability

Molecular Weight	505.69	Formula	C₃₂H₄₃NO₄	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	218600-53-4	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	RTA 402, TP-155, NSC 713200, CDDO Methyl Ester, CDDO-Me			Smiles	CC1(CCC2(CCC3(C(C2C1)C(=O)C=C4C3(CCC5C4(C=C(C(=O)C5(C)C)C#N)C)C)C)C(=O)OC)C

Mechanism of Action

Features	The only IKKβ inhibitor in clinical use for solid tumors, type 2 diabetes, and chronic kidney disease. An orally-available antioxidant inflammation modulator.
Targets/IC₅₀/Ki	IKK (Cell-free assay) Ferroptosis Nrf2 NF-κB
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. This compound decreases the viability of leukemic HL-60, KG-1, and NB4 cells with IC50 of 0.4, 0.4, and 0.27 μM, respectively. It induces pro-apoptotic Bax protein, inhibits the activation of ERK1/2, and it blocks Bcl-2 phosphorylation, which contributes to the induction of apoptosis. This chemical potently inhibits both constitutive and inducible NF-kappaB activated by TNF, interleukin (IL)-1beta, phorbol ester, okadaic acid, hydrogen peroxide, lipopolysaccharide, and cigarette smoke.
Kinase Assay	IKK assay
Kinase 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.
In vivo	Bardoxolone Methyl (60 mg/kg) reduces the number, size, and severity of lung tumors in vivo. This compound 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.
References	[1] https://pubmed.ncbi.nlm.nih.gov/11063620/ [2] https://pubmed.ncbi.nlm.nih.gov/11756188/ [3] https://pubmed.ncbi.nlm.nih.gov/16551868/ [4] https://pubmed.ncbi.nlm.nih.gov/17363558/ [5] https://pubmed.ncbi.nlm.nih.gov/20626291/ [6] https://pubmed.ncbi.nlm.nih.gov/20234191/ [7] https://pubmed.ncbi.nlm.nih.gov/31541463/

Applications

Methods	Biomarkers	PMID
Western blot	p-IκBα / IκBα Bcl-xl / Bcl-2 / Bax / Cleaved caspase / Cytochrome C / PARP / Cleaved PARP p-PI3K / PI3K / p-AMPK / AMPK / p-p38 MAPK / p38 MAPK / p-AKT / AKT / p-mTOR / mTOR PTEN / PP2A / PHLPP1	25897966
Immunofluorescence	PDI / SDHA c-PARP / Cytochrome C / COX IV	26053096
Growth inhibition assay	Cell viability	25733817

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT02316821	Completed	Chronic Kidney Disease\|Type 2 Diabetes	Kyowa Kirin Co. Ltd.	December 2014	Phase 2
NCT02036970	Completed	Pulmonary Arterial Hypertension\|Pulmonary Hypertension\|Interstitial Lung Disease\|Idiopathic Interstitial Pneumonia\|Idiopathic Pulmonary Fibrosis\|Sarcoidosis\|Respiratory Bronchiolitis Associated Interstitial Lung Disease\|Desquamative Interstitial Pneumonia\|Cryptogenic Organizing Pneumonia\|Acute Interstitial Pneumonitis\|Idiopathic Lymphoid Interstitial Pneumonia\|Idiopathic Pleuroparenchymal Fibroelastosis	Reata a wholly owned subsidiary of Biogen\|Biogen	May 31 2014	Phase 2
NCT01598363	Completed	Healthy Volunteers	Reata a wholly owned subsidiary of Biogen\|Biogen	June 30 2012	Phase 1
NCT01551446	Withdrawn	Renal Insufficiency Chronic\|Diabetes Mellitus Type 2	Reata a wholly owned subsidiary of Biogen\|Biogen	April 30 2012	Phase 1
NCT01503866	Completed	Healthy	Reata a wholly owned subsidiary of Biogen\|Biogen	December 1 2011	Phase 1

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