SAR study of celastrol analogs targeting Nur77-mediated inflammatory pathway

Chen, Ziwen; Zhang, Duo; Yan, Siwei; Hu, Chaochao; Huang, Zhenfei; Li, Zhuoer; Peng, Shuangzhou; Li, Xiaotong; Zhu, Yi; Yu, Hongyu; Lian, Baohuan; Kang, Qi; Li, Mingyu; Zeng, Zhiping; Zhang, Xiaokun; Su, Ying

doi:10.1016/j.ejmech.2019.05.009

Cited by 32 publications

(12 citation statements)

References 32 publications

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“…Therefore, our wound-healing experiment was done in a narrow time-window, this protocol had precluded the effect of proliferation. Since the binding affinity of celastrol to ROCK2 is confirmed, the SAR optimization 52 is needed for translational application of celastrol. Currently, we are establishing hepatocyte organoids model so we can model the HCC progression and screen for potent compounds derived from celastrol.…”

Section: Discussionmentioning

confidence: 99%

Celastrol inhibits ezrin-mediated migration of hepatocellular carcinoma cells

Song

et al. 2020

Sci Rep

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Chemicals and antibody. Celastrol was ordered from MedChemExpress under the item number HY-13067. Antibodies against ezrin, ezrin pT567 and α-tubulin (DM1A) were purchased from Cell Signaling Technology. Antibodies against ROCK2 and ROCK2 (phospho S1366) were purchased from Abcam. Anti-GFP antibodies were purchased from Santa Cruz Biotechnology. Anti-FLAG-tag (M2) antibody was from Sigma. plasmids. The development of bacterial expression vectors containing human ezrin fused to histidine was described previously 61. EGFP-tagged ezrin plasmids was produced as described before 62. PCR amplified ROCK2 cDNA was cloned into the 3× FLAG-Myc-CMV-24 vector (Sigma) by BamHI and RcoRI digestion. The mutanted of ezrin and ROCK2 were constructed by Fast Mutagenesis Kit (Vazyme Biotech). All plasmids were verified by sequencing (Tsingke Biological Tech). cell culture and transfection. MHCC97H was a gift from Professor Yong Chen (Fourth Military Medical University) 20. HepG2 and HEK293T cells were from American Type Culture Collection (ATCC). Huh7 cells were from National Infrastructure of Cell Line Resource. The cells were incubated according to ATCC instruction. Cells were transfected using conventional calcium phosphate method or the Lipofectamine 3,000 (Invitrogen) according to manufacturer's instructions. Determination of cell viability (MTS assay). Celastrol cytotoxicity was assessed with the use of a CellTiter 96 AQueous One Solution Cell Proliferation Assay (Promega), which is a form of the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay. Cells (5,000 cells/well) were plated in 96-well plates, treated with various concentrations of celastrol for 24 h. After celastrol treatment, cell viability was determined by adding a small amount of the One Solution Reagent directly to culture wells, incubating for 3 h and then recording the absorbance at 490 nm with a 96-well plate reader. Wound healing assay. MHCC97H, HepG2 or Huh7 cells with logarithmic growth phases, was cultivated in 12 orifices with 5 × 10 5 cells density. Each group has three replicates. After a starvation of 6 h in serum-free medium, 10 μl Tip was taken to vertical scratch "#" glyph at the bottom of the culture plate. The floating cells were washed off with PBS and the celastrol was dissolved in medium supplemented with 20% FBS. Samples were photographed at 0 h, 4 h and 8 h. Image J software was used to measure and compare the effects of celastrol on cell migration. Single cell tracking assay. The cell migration by single cell tracking assay was performed as previously described 32. Briefly, MHCC97H cells were laid in a Silian dish. After stabilized, the cells were starved with Opti-MEM for 6 h, and then cultured in DMEM containing 20% FBS. The movement track of 10 single-cells in each field of view were recorded by microscope for 6 h, with 10 min interval at once.

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Section: Discussionmentioning

confidence: 99%

Celastrol inhibits ezrin-mediated migration of hepatocellular carcinoma cells

Song

et al. 2020

Sci Rep

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“…Moreover, many of the 3,5-disubstituted analogs of DIM-C-pPhOH inhibit tumor growth in mouse xenograft models at doses < 5 mg/kg per day and ongoing studies have identified ligands that inhibit tumor growth at doses < 1mg/kg per day. Interestingly, several other classes of compounds bind and modulate NR4A1-dependent pathways/genes and these include cytosporone B and structurally related compounds, IMCA (2-imino-6-methoxy-2H-chromene-3-carbothiomide) and the triterpenoid celastrol and analogs[ 76 - 80 ] (Figure 5 ). Fascinatingly, celastrol also activates ROS-dependent downregulation of Sp1, Sp3 and Sp4 in bladder cancer cells[ 80 ] ( e.g.…”

Section: Drugs That Target Sp Tfs and Nr4a1mentioning

confidence: 99%

“…Interestingly, several other classes of compounds bind and modulate NR4A1-dependent pathways/genes and these include cytosporone B and structurally related compounds, IMCA (2-imino-6-methoxy-2H-chromene-3-carbothiomide) and the triterpenoid celastrol and analogs[ 76 - 80 ] (Figure 5 ). Fascinatingly, celastrol also activates ROS-dependent downregulation of Sp1, Sp3 and Sp4 in bladder cancer cells[ 80 ] ( e.g. , Figure 4 ) and uniquely acts as a bifunctional agent targeting both Sp TFs and NR4A1.…”

Section: Drugs That Target Sp Tfs and Nr4a1mentioning

confidence: 99%

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Transcription factors specificity protein and nuclear receptor 4A1 in pancreatic cancer

Safe¹,

Shrestha²,

Mohankumar³

et al. 2021

WJG

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Specificity protein (Sp) transcription factors (TFs) Sp1, Sp3 and Sp4, and the orphan nuclear receptor 4A1 (NR4A1) are highly expressed in pancreatic tumors and Sp1 is a negative prognostic factor for pancreatic cancer patient survival. Results of knockdown and overexpression of Sp1, Sp3 and Sp4 in pancreatic and other cancer lines show that these TFs are individually pro-oncogenic factors and loss of one Sp TF is not compensated by other members. NR4A1 is also a pro-oncogenic factor and both NR4A1 and Sp TFs exhibit similar functions in pancreatic cancer cells and regulate cell growth, survival, migration and invasion. There is also evidence that Sp TFs and NR4A1 regulate some of the same genes including survivin, epidermal growth factor receptor, PAX3-FOXO1, α5- and α6-integrins, β1-, β3- and β4-integrins; this is due to NR4A1 acting as a cofactor and mediating NR4A1/Sp1/4-regulated gene expression through GC-rich gene promoter sites. Several studies show that drugs targeting Sp downregulation or NR4A1 antagonists are highly effective inhibitors of Sp/NR4A1-regulated pathways and genes in pancreatic and other cancer cells, and the triterpenoid celastrol is a novel dual-acting agent that targets both Sp TFs and NR4A1.

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“…Chen's lab verified that celastrol binds to Nur77 and inhibits inflammation in vitro 93 . To further study the SAR of celastrol derivatives in relation to the proinflammatory pathway, Chen et al 94 designed and synthesized 24 celastrol analogs by modifying the carboxylic acid or hydroxyl groups (Scheme 13). The fluorescence quenching assay was used to evaluate their Nur77‐binding affinity, whereas their anti‐inflammatory effects were evaluated by detecting the level of IkBa in tumor necrosis factor‐α (TNF‐α)‐treated HepG2 cells.…”

Section: Chemical Synthesis and Structural Modificationsmentioning

confidence: 99%

Biosynthesis, total synthesis, structural modifications, bioactivity, and mechanism of action of the quinone‐methide triterpenoid celastrol

Lü

Liu

Zhou

et al. 2020

Medicinal Research Reviews

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Celastrol, a quinone‐methide triterpenoid, was extracted from Tripterygium wilfordii Hook. F. in 1936 for the first time. Almost 70 years later, it is considered one of the molecules most likely to be developed into modern drugs, as it exhibits notable bioactivity, including anticancer and anti‐inflammatory activity, and exerts antiobesity effects. In addition, the molecular mechanisms underlying its bioactivity are being widely studied, which offers new avenues for its development as a pharmaceutical reagent. Owing to its potential therapeutic effects and unique chemical structure, celastrol has attracted considerable interest in the fields of organic, biosynthesis, and medicinal chemistry. As several steps in the biosynthesis of celastrol have been revealed, the mechanisms of key enzymes catalyzing the formation and postmodifications of the celastrol scaffold have been gradually elucidated, which lays a good foundation for the future heterogeneous biosynthesis of celastrol. Chemical synthesis is also an effective approach to obtain celastrol. The total synthesis of celastrol was realized for the first time in 2015, which established a new strategy to obtain celastroid natural products. However, owing to the toxic effects and suboptimal pharmacological properties of celastrol, its clinical applications remain limited. To search for drug‐like derivatives, several structurally modified compounds were synthesized and tested. This review focuses primarily on the latest research progress in the biosynthesis, total synthesis, structural modifications, bioactivity, and mechanism of action of celastrol. We anticipate that this paper will facilitate a more comprehensive understanding of this promising compound and provide constructive references for future research in this field.

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SAR study of celastrol analogs targeting Nur77-mediated inflammatory pathway

Cited by 32 publications

References 32 publications

Celastrol inhibits ezrin-mediated migration of hepatocellular carcinoma cells

Celastrol inhibits ezrin-mediated migration of hepatocellular carcinoma cells

Transcription factors specificity protein and nuclear receptor 4A1 in pancreatic cancer

Biosynthesis, total synthesis, structural modifications, bioactivity, and mechanism of action of the quinone‐methide triterpenoid celastrol

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