solo para uso en investigación

Bardoxolone Methyl (RTA 402) Activador de NRF2

Cat. No.S8078

Bardoxolone Methyl (RTA 402, TP-155, NSC 713200, CDDO Methyl Ester, CDDO-Me) es un inhibidor de la IKK, mostrando potentes actividades proapoptóticas y antiinflamatorias; también un potente activador de Nrf2 y un inhibidor del factor nuclear-κB (NF-κB). Bardoxolone Methyl abroga la ferroptosis. El metil bardoxolona induce apoptosis y autofagia en células cancerosas.
Bardoxolone Methyl (RTA 402) Nrf2 activador Chemical Structure

Estructura química

Peso molecular: 505.69

Saltar a

Control de calidad

Lote: Pureza: 99.86%
99.86

Cultivo celular, tratamiento y concentración de trabajo

Líneas celulares Tipo de ensayo Concentración Tiempo de incubación Formulación Descripción de la actividad 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
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Información química, almacenamiento y estabilidad

Peso molecular 505.69 Fórmula

C32H43NO4

Almacenamiento (Desde la fecha de recepción)
Nº CAS 218600-53-4 Descargar SDF Almacenamiento de soluciones madre

Sinónimos 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

Solubilidad

In vitro
Lote:

DMSO : 26 mg/mL (51.41 mM)
(El DMSO contaminado con humedad puede reducir la solubilidad. Usar DMSO fresco y anhidro.)

Water : Insoluble

Ethanol : Insoluble

Calculadora de Molaridad

Masa Concentración Volumen Peso molecular
Calculadora de Dilución Calculadora de Peso Molecular

In vivo
Lote:

Calculadora de formulación in vivo (Solución clara)

Paso 1: Introduzca la información a continuación (Recomendado: Un animal adicional para tener en cuenta la pérdida durante el experimento)

mg/kg g μL

Paso 2: Introduzca la formulación in vivo (Esto es solo la calculadora, no la formulación. Por favor, contáctenos primero si no hay una formulación in vivo en la sección de Solubilidad.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Resultados del cálculo:

Concentración de trabajo: mg/ml;

Método para preparar el líquido maestro de DMSO: mg fármaco predissuelto en μL DMSO ( Concentración del líquido maestro mg/mL, Por favor, contáctenos primero si la concentración excede la solubilidad del DMSO del lote del fármaco. )

Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadirμL PEG300, mezclar y clarificar, luego añadirμL Tween 80, mezclar y clarificar, luego añadir μL ddH2O, mezclar y clarificar.

Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadir μL Aceite de maíz, mezclar y clarificar.

Nota: 1. Por favor, asegúrese de que el líquido esté claro antes de añadir el siguiente disolvente.
2. Asegúrese de añadir el (los) disolvente(s) en orden. Debe asegurarse de que la solución obtenida, en la adición anterior, sea una solución clara antes de proceder a añadir el siguiente disolvente. Se pueden utilizar métodos físicos como el vórtice, el ultrasonido o el baño de agua caliente para ayudar a la disolución.

Mecanismo de acción

Características
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

El Bardoxolone Methyl exhibe potentes actividades inhibitorias contra la producción de óxido nítrico inducida por interferón-Ƴ en macrófagos de ratón con una IC50 de 0.1 nM. Este compuesto disminuye la viabilidad de las células leucémicas HL-60, KG-1 y NB4 con una IC50 de 0.4, 0.4 y 0.27 μM, respectivamente. Induce la proteína pro-apoptótica Bax, inhibe la activación de ERK1/2 y bloquea la fosforilación de Bcl-2, lo que contribuye a la inducción de la apoptosis. Este químico inhibe potentemente la NF-kappaB constitutiva e inducible activada por TNF, interleucina (IL)-1beta, éster de forbol, ácido okadaico, peróxido de hidrógeno, lipopolisacárido y humo de cigarrillo.

Ensayo de quinasa
Ensayo de IKK
Para determinar el efecto de CDDO-Me en la activación de IKK inducida por TNF, se analiza IKK. Brevemente, el complejo IKK de extractos de células enteras se precipitó con un anticuerpo contra IKKα e IKKβ y luego se trató con perlas de proteína A/G-Sepharose. Después de 2 horas, las perlas se lavan con tampón de lisis y luego se resuspenden en una mezcla de ensayo de quinasa que contiene 50 mmol/L HEPES (pH 7.4), 20 mmol/L MgCl2, 2 mmol/L DTT, 20 μCi [γ-32P]ATP, 10 μmol/L ATP no marcado y 2 μg del sustrato glutatión S-transferasa-IκBα (aminoácidos 1-54). Después de la incubación a 30°C durante 30 minutos, la reacción se termina hirviendo con tampón de muestra SDS durante 5 minutos. Finalmente, la proteína se resuelve en SDS-PAGE al 10%, el gel se seca y las bandas radiactivas se visualizan con un Storm820. Para determinar las cantidades totales de IKK-α e IKK-β en cada muestra, 50 μg de proteínas de células enteras se resuelven en SDS-PAGE al 7.5%, se transfieren electroforéticamente a una membrana de nitrocelulosa y luego se blotean con anticuerpos anti-IKK-α o anti-IKK-β.
In vivo

El Bardoxolone Methyl (60 mg/kg) reduce el número, tamaño y gravedad de los tumores pulmonares in vivo. Este compuesto reduce significativamente la respuesta de citoquinas inflamatorias in vivo después del desafío con LPS, induce la expresión de la proteína HO-1 en el bazo y protege a los ratones contra una dosis letal de LPS.

Referencias
  • [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/

Aplicaciones

Métodos Biomarcadores Imágenes 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
S8078-WB1
25897966
Immunofluorescence PDI / SDHA c-PARP / Cytochrome C / COX IV
S8078-IF1
26053096
Growth inhibition assay Cell viability
S8078-viability1
25733817

Información del ensayo clínico

(datos de https://clinicaltrials.gov, actualizado el 2024-05-22)

Número NCT Reclutamiento Condiciones Patrocinador/Colaboradores Fecha de inicio Fases
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

Soporte técnico

Instrucciones de manipulación

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