Bardoxolone primarily targets the Nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, functioning as a potent Nrf2 activator . It blocks the synthesis of inducible nitric oxide synthase (iNOS) and inducible cyclooxygenase (COX-2), two enzymes involved in inflammation and carcinogenesis . Additionally, bardoxolone inhibits interleukin-1 (IL-1)-induced expression of the pro-inflammatory proteins matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-13 (MMP-13), as well as the expression of Bcl-3, an IL-1-responsive gene that contributes to MMP-1 expression . The compound also inhibits Janus-activated kinase/STAT signaling and NF-κB pathways . The methyl ester form (Bardoxolone methyl) disrupts the Keap1-Nrf2 interaction, activating Nrf2 signaling .

Novel synthetic triterpenoid, bardoxolone methyl, is an antioxidant inflammation modulator that potently induces Nrf2 and inhibits Janus-activated kinase/STAT signaling, NF-κB. It has been demonstrated that bardoxolone methyl causes cancer cell lines to differentiate, stop growing, and undergo apoptosis[2].

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Bardoxolone has been shown to induce differentiation, inhibit proliferation, and induce apoptosis in cancer cell lines . It causes cancer cell lines to undergo programmed cell death, particularly in cells under high levels of intrinsic oxidative stress . In contrast, in normal cells, bardoxolone induces protective antioxidant and anti-inflammatory responses . For the methyl ester form (Bardoxolone methyl), in vitro studies demonstrate that it potently induces Nrf2 and inhibits Janus-activated kinase/STAT signaling and NF-κB . Bardoxolone methyl reduces the viability of leukemia HL-60, KG-1, and NB4 cells with IC50 values of 0.4, 0.4, and 0.27 μM, respectively . It also induces pro-apoptotic Bax protein expression, inhibits ERK1/2 activation, and suppresses Bcl-2 phosphorylation, contributing to apoptosis induction .

Although the groups' mRNA expression is similar, kidney sections from the bardoxolone methyl-treated monkeys show reduced megalin protein expression. Densitometry analyses validate the observed reduction in megalin protein expression, revealing a significant decrease in megalin protein expression in the monkey kidney upon administration of Bardoxolone methyl. Both the mRNA and protein expression of cubilin in the kidney are unaffected by the administration of bardoxolone methyl. On day 28, the creatinine clearance of monkeys given Bardoxolone methyl was considerably different from that of baseline and animals given a vehicle treatment. Urinary albumin-to-creatinine ratios (UACRs), as measured from 24-hour urine collections, are significantly higher after 28 days of Bardoxolone methyl administration in comparison to animals receiving vehicle. Notably, UACRs rose by 27.9% in monkeys treated with bardoxolone methyl and decreased by 53.3% in animals treated with a vehicle[3]. For 21 weeks, male C57BL/6J mice are either fed a low-fat diet (LFD), only a high-fat diet (HFD), or oral BARD during HFD feeding (HFD/BARD). HFD mice show a significant increase in F4/80 crown-like structures (+95%; p<0.001) in comparison to LFD mice, which is effectively prevented by BARD (−50%; p<0.01). Similarly, compared to LFD mice and HFD/BARD mice, the number of F4/80 interstitial macrophages in HFD mice is significantly higher by 98% (p<0.001) and 32% (p<0.01), respectively[4].

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In vivo studies have demonstrated significant single-agent anti-cancer activity of bardoxolone methyl in preclinical models . In a 12-month GLP study in cynomolgus monkeys, bardoxolone methyl was orally administered at doses of 5, 30, and 300 mg/kg once daily, with interim analysis at 6 months and post-dose recovery analysis 4 weeks after the final dose . In a 28-day monkey study, female cynomolgus monkeys received bardoxolone methyl at 30 mg/kg per day, and the treatment resulted in decreased megalin protein expression in the kidney and significantly altered creatinine clearance compared to baseline . In high-fat diet mouse models, oral administration of bardoxolone (10 mg/kg in drinking water for 21 weeks) effectively prevented the increase in F4/80 crown-like structures (by 50%) and reduced the number of F4/80 interstitial macrophages compared to HFD controls, demonstrating anti-inflammatory effects in adipose tissue .

For the methyl ester form (Bardoxolone methyl), the disruption of Keap1-Nrf2 interaction was measured using an in vitro Keap1-Nrf2 binding assay, demonstrating an IC50 of 1.2 ± 0.1 μM . The TNF-α-induced NF-κB reporter gene assay in HEK293T cells showed that Bardoxolone methyl inhibits NF-κB activation with an IC50 of 3.5 ± 0.3 μM . Additionally, a NO production assay demonstrated that Bardoxolone methyl suppresses iNOS activity in activated macrophages with an IC50 of 2.8 ± 0.2 μM . These biochemical assays are typically performed using recombinant proteins or cell lysates in cell-free systems to quantify the compound's direct binding or inhibitory effects on specific molecular targets.

In vitro cell-based assays for bardoxolone typically involve culturing cancer cell lines or primary cells in appropriate media and treating them with various concentrations of the compound. For cell viability assessment, cells are seeded in 96-well plates and treated with bardoxolone (or its methyl ester form) at concentrations ranging from nanomolar to micromolar levels for 24-72 hours, followed by standard viability assays such as MTT or ATP quantification to calculate IC50 values . Apoptosis is evaluated by measuring pro-apoptotic protein expression (e.g., Bax) via Western blotting, assessing ERK1/2 activation and Bcl-2 phosphorylation status, or using flow cytometry with Annexin-V staining . For Nrf2 activation studies, cells transfected with ARE-LUC reporter constructs are treated with bardoxolone for 24 hours, and relative luminescence values are measured to quantify pathway activation .

Monkey studies: In a 12-month GLP study, cynomolgus monkeys (n=9 per sex/dose group) were orally administered bardoxolone methyl at 5, 30, and 300 mg/kg once daily for 12 months, with an interim analysis at 6 months and a postdose recovery analysis 4 weeks after the final dose . In a 28-day study, female cynomolgus monkeys (n=6 for vehicle, n=12 for treatment) received bardoxolone methyl at 30 mg/kg per day once daily for 28 days . Mouse studies: Male C57BL/6J mice were divided into three groups (n=7): low-fat diet control, high-fat diet (40% fat), and high-fat diet supplemented with bardoxolone in drinking water at 10 mg/kg body weight for 21 weeks. All mice were euthanized using CO2 asphyxiation, and mesenteric fat tissue was collected fo

Specific pharmacokinetic data for bardoxolone (RTA 401) as a single agent are not detailed in the available literature. However, the methyl ester form (bardoxolone methyl, RTA 402) has been characterized in clinical studies. In a Phase I trial of patients with advanced solid tumors and lymphoid malignancies, bardoxolone methyl was administered orally for the first 21 days of a 28-day cycle . Pharmacokinetic characterization was a primary outcome measure, with plasma samples collected to determine drug concentration parameters . The compound is formulated as capsules for oral administration, taken once daily in the morning prior to food intake . Solubility data indicate that bardoxolone is soluble in DMSO at ~100 mg/mL (~203.39 mM), and in vivo formulations can be prepared using 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline or 10% DMSO + 90% corn oil .

Bardoxolone methyl has been associated with significant safety concerns in clinical trials. The Phase 3 BEACON trial evaluating bardoxolone methyl for chronic kidney disease was terminated prematurely in October 2012 after patients treated with the drug experienced a higher rate of heart-related adverse events, including heart failure, hospitalizations, and deaths . Post-hoc analyses revealed that this severe adverse event was likely driven by acute fluid and sodium retention, particularly in patients with pre-existing cardiovascular risk factors . In a Phase I clinical trial, common grade 3/4 treatment-related adverse events included thrombocytopenia, lymphocytopenia, neutropenia, and anemia . Despite these cardiovascular concerns, the compound continues to be studied for oncology indications due to its dual mechanisms of Nrf2 activation and NF-κB inhibition .

[1]. Cardiac and renal function in patients with type 2 diabetes who have chronic kidney disease: potential effectsof bardoxolone methyl. Drug Des Devel Ther. 2012;6:141-9.
[2]. A phase I first-in-human trial of bardoxolone methyl in patients with advanced solid tumors and lymphomas. Clin Cancer Res. 2012 Jun 15;18(12):3396-406.
[3]. Bardoxolone Methyl Decreases Megalin and Activates Nrf2 in the Kidney. J Am Soc Nephrol. 2012 Oct;23(10):1663-73.
[4]. Bardoxolone Methyl Prevents Mesenteric Fat Deposition and Inflammation in High-Fat Diet Mice. ScientificWorldJournal. 2015;2015:549352

Badoxuron belongs to the cyclohexenone class of compounds. Badoxuron has been used in clinical trials for the treatment of lymphoma and solid tumors. It is a synthetic triterpenoid compound that efficiently activates redox-sensitive signaling pathways, inducing programmed cell death (apoptosis) in cancer cells under high levels of endogenous oxidative stress. Conversely, badoxuron induces a protective antioxidant/anti-inflammatory response in normal cells. Badoxuron is a synthetic triterpenoid compound with potential antitumor and anti-inflammatory activities. Badoxuron blocks the synthesis of inducible nitric oxide synthase (iNOS) and inducible cyclooxygenase (COX-2), both of which are involved in inflammation and carcinogenesis. The drug also inhibits the expression of interleukin-1 (IL-1)-induced pro-inflammatory proteins matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-13 (MMP-13), as well as Bcl-3 expression; Bcl-3 is an IL-1-responsive gene that preferentially promotes MMP-1 gene expression.

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Mechanism of Action
Badosulone is a synthetic triterpenoid compound that efficiently activates redox-sensitive signaling pathways, inducing programmed cell death (apoptosis) in cancer cells under high levels of endogenous oxidative stress. Conversely, bardosulone induces protective antioxidant/anti-inflammatory responses in normal cells. Numerous studies in human cancer animal models have demonstrated that bardosulone is a potent anticancer drug with a proven ability to inhibit tumor growth and promote tumor regression, whether used alone or in combination with radiotherapy and chemotherapy. At dose levels that produce anticancer effects, bardosulone also inhibits radiation- and chemotherapy-induced damage to normal tissues (e.g., oral mucositis). Bardosulone induces apoptosis through caspase-dependent and caspase-independent mechanisms, the latter involving caspase-8 activation, Bid cleavage, cytochrome c release, and caspase-3 activation. Furthermore, the JNK, p38, and ERK pathways are involved in bardosulone-induced apoptosis in tumor cell lines, a process mediated by intracellular redox homeostasis involving decreased glutathione and increased reactive oxygen species. Studies have shown that bardosolone enhances the expression of p42 CEBPA protein at the translational level.

C31H41NO4

491.672

491.303

218600-44-3

218600-53-4

400010

White to yellow solid powder

1.2±0.1 g/cm3

632.9±55.0 °C at 760 mmHg

336.6±31.5 °C

0.0±4.0 mmHg at 25°C

1.575

5.76

1

5

1

36

1200

7

C[C@@]12CC[C@]3(CCC(C[C@H]3[C@H]1C(=O)C=C4[C@]2(CC[C@@H]5[C@@]4(C=C(C(=O)C5(C)C)C#N)C)C)(C)C)C(=O)O

TXGZJQLMVSIZEI-UQMAOPSPSA-N

InChI=1S/C31H41NO4/c1-26(2)10-12-31(25(35)36)13-11-30(7)23(19(31)16-26)20(33)14-22-28(5)15-18(17-32)24(34)27(3,4)21(28)8-9-29(22,30)6/h14-15,19,21,23H,8-13,16H2,1-7H3,(H,35,36)/t19-,21-,23-,28-,29+,30+,31-/m0/s1

(4aS,6aR,6bS,8aR,12aS,14aR,14bS)-11-cyano-2,2,6a,6b,9,9,12a-heptamethyl-10,14-dioxo-1,3,4,5,6,7,8,8a,14a,14b-decahydropicene-4a-carboxylic acid

Bardoxolone; CDDO; RTA-401; RTA 401; RTA401

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~203.39 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.08 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (5.08 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)