Oridonin

Manufactured by Merck Group

Sourced in United States

Oridonin is a natural compound extracted from the Chinese herbal plant Isodon. It is a white crystalline powder that is commonly used in laboratory research applications. Oridonin exhibits various biological activities and is often employed as a research tool in the study of cellular processes and signaling pathways.

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Lab products found in correlation

Trypsin, by Thermo Fisher Scientific (5 mentions) Dimethyl sulfoxide (dmso), by Merck Group (4 mentions) Propidium iodide, by MP Biomedicals (3 mentions) Phospho smad2 3, by Cell Signaling Technology (2 mentions) Anti p53, by Cell Signaling Technology (2 mentions) Antibody against α smooth muscle actin α sma, by Merck Group (2 mentions) Antibodies against fibronectin sc 6952, by Santa Cruz Biotechnology (2 mentions) Cleaved parp, by Cell Signaling Technology (2 mentions) Cleaved caspase 9, by Cell Signaling Technology (2 mentions) Anti p21, by BD (2 mentions) Anti collagen type 1 polyclonal antibody 600 401 103, by Rockland Immunochemicals (2 mentions) Gapdh antibody 10r g109a, by Biosynth (2 mentions) Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions) Gemcitabine hydrochloride, by Selleck Chemicals (1 mentions) Imatinib, by Merck Group (1 mentions) Anti shp2, by Santa Cruz Biotechnology (1 mentions) Lipofectamine ltx reagent, by Thermo Fisher Scientific (1 mentions) Anti p stat5, by Cell Signaling Technology (1 mentions) Anti myc tag, by Cell Signaling Technology (1 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)

Close spelling variants of this product

Oridonin (C20H28O6)

20 protocols using oridonin

Oridonin Compound Cytotoxicity Assay

Cited 2 times

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oridonin compound was purchased from Beijing OKA Biological Technology Co., Ltd. (Beijing, China), and the purity was confirmed to be ≥98%. The oridonin solution was prepared in DMSO (Sigma, MO, USA). Gemcitabine hydrochloride was purchased from Selleck Chemical (Houston, TX, USA) and made up in sterile phosphate-buffered saline (PBS) with the stock solution of 160 mg/kg.
Seed cells on 96-well plates by 5000 cells/well and treat with oridonin, with, as indicated, final concentrations for a different duration. Fix cells in situ with 40% (w/v) cold Trichloroacetic acid (TCA). Let them sit for 5 min. Incubate plates at 4 °C for 1 h. Discard supernatant and gently wash plates 5 times with water, followed by an air-dry. Stain fixed cells with 0.4% (w/v) SRB with 1% acetic acid as solubilizer. After staining, wash with 1% acetic acid to remove the free dye and air-dry. Solubilize the bound stain with 100 mM Tris Base for reading. Read absorbance on a microplate reader at 515 nm [45 (link),46 (link)]. Inhibition% = [(OD _control − OD _treated)]/OD _control] × 100%.

Zhang X., Xing M., Ma Y., Zhang Z., Qiu C., Wang X., Zhao Z., Ji Z, & Zhang J.Y. (2023). Oridonin Induces Apoptosis in Esophageal Squamous Cell Carcinoma by Inhibiting Cytoskeletal Protein LASP1 and PDLIM1. Molecules, 28(2), 805.

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Investigating Anti-Abl Signaling Pathway

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SBF‐1 is a synthetic steroidal glycoside, as described previously.¹⁵, ¹⁶ Anti‐c‐Abl, anti‐p‐Bcr, anti‐p‐STAT5, anti‐STAT5, anti‐p‐SHP‐2, anti‐c‐Cbl, anti‐HA‐tag, anti‐myc‐tag, and anti‐Beclin 1 antibodies were purchased from Cell Signaling Technology. Anti‐SHP‐2, anti‐GAPDH, anti‐PTP1B, and anti‐ ubiquitin antibodies were from Santa Cruz Biotechnology. Anti‐p62, anti‐ATG5, and anti‐phosphotyrosine antibodies were from Abcam. MG132 and bafilomycin A1 (Baf A1) were from Selleck Chemicals. Anti‐Flag‐tag antibody, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT), imatinib, 3‐Methyladenine (3‐MA), chloroquine (CQ), 4′,6‐diamidino‐2‐phenylindole (DAPI), and Oridonin were from Sigma‐Aldrich. The lysosome‐specific dye LysoTraker Red, Lipofectamine™ LTX Reagent, Lipofectamine 2000, and Lipofectamine RNAi MAX were from Life Technologies. SuperSep Phos‐tag™ was from Wako. The RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit was from Thermo Fisher Scientific.

Elgehama A., Wang Y., Yu Y., Zhou L., Chen Z., Wang L., Sun L., Gao J., Yu B., Shen Y, & Xu Q. (2022). Targeting the PTP1B‐Bcr‐Abl1 interaction for the degradation of T315I mutant Bcr‐Abl1 in chronic myeloid leukemia. Cancer Science, 114(1), 247-258.

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Oridonin Ameliorates Liver Fibrosis

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Oridonin was obtained from Sigma‐Aldrich [Purity 98.0% (HPLC), St. Louis, MO]. Carbon tetrachloride (CCl₄) and corn oil were purchased from Aladdin (Shanghai, China). Antibodies against α‐SMA and β‐actin were from BOSTER (Wuhan, China) and TransGen (Beijing, China), respectively; horseradish peroxidase (HRP)‐conjugated Affinity Pure goat anti‐mouse IgG and anti‐rabbit IgG were purchased from Zhongshan Golden Bridge Biotechnology Co, Ltd. (Beijing, China). Enhanced chemiluminescence detection reagents were purchased from Clinx (Shanghai, China) and the chemiluminescence system was obtained from New Life Science Products (Boston, MA). Alanine aminotransferase (ALT) and hydroxyproline assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Liu D., Qin H., Yang B., Du B, & Yun X. (2020). Oridonin ameliorates carbon tetrachloride‐induced liver fibrosis in mice through inhibition of the NLRP3 inflammasome. Drug Development Research, 81(4), 526-533.

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Oridonin Cytotoxicity in 4T1 Breast Cancer

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Oridonin (purity ≥ 98%) was purchased from Sigma-Aldrich China Inc. (Shanghai, China). The human breast cancer cell line 4T1 were obtained from the American Type Culture Collection (Rockville, Maryland, USA). Cell counting kit-8 (CCK-8) was provided by Dojindo Laboratories (Kumamoto, Japan). Hoechst 33258, the ApoDETECT annexin V–fluorescein isothiocyanate (FITC) apoptosis detection kit, the Light Shift chemiluminescent electrophoretic mobility shift assay (EMSA) kit, and NE-PER nuclear and cytoplasmic extraction reagents were obtained from ThermoFisher Scientific Inc. (Waltham, Massachusetts, USA). The cells were cultured in DMEM medium (Hyclone, Beijing, China) enriched with 10% fetal bovine serum (Hyclone) at 37 °C and 5% CO₂.

Xia S., Zhang X., Li C, & Guan H. (2017). Oridonin inhibits breast cancer growth and metastasis through blocking the Notch signaling. Saudi Pharmaceutical Journal : SPJ, 25(4), 638-643.

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Synthesis and Evaluation of CYD0682

Cited 1 time

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All cell culture mediums and trypsin were purchased from Life Technology Corp. (Carlsbad, CA). Oridonin was purchased from Sigma-Aldrich Co. LLC. (St. Louis, MO). Transforming growth factor β1 (TGF-β) was purchased from R&D Systems Inc. (Minneapolis, MN). Propidium iodide was purchased from MP Biomedicals, LLC (Solon, OH). HOESCHT 33342 (Cat#83218) was purchased from AnaSpec Inc.(Fremont, CA). CYD0682 is a novel analogue of 1-ene designed from Oridonin by removal of 1-hydroxyl group and introduction of a double bond in the A-ring (Figure 1). CYD0682 was synthesized following our previously reported protocols [14 (link)].

Bohanon F.J., Wang X., Graham B.M., Ding C., Ding Y., Rastellini C., Zhou J, & Radhakrishnan R.S. (2015). Enhanced Effects of Novel Oridonin Analog CYD0682 for Hepatic Fibrosis. The Journal of surgical research, 199(2), 441-449.

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GC and HEK293 Cell Culture

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The human GC cell line SNU-216 and human kidney epithelial cell line HEK293 were purchased from the American Type Culture Collection (ATCC, USA). Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco, USA) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco). The cells were seeded onto culture dishes at 37°C in a humidified 5% CO₂ incubator. Oridonin (Sigma-Aldrich, USA) was dissolved in DMSO and diluted into adequate volume of DMEM.

Bi E., Liu D., Li Y., Mao X., Wang A, & Wang J. (2018). Oridonin induces growth inhibition and apoptosis in human gastric carcinoma cells by enhancement of p53 expression and function. Brazilian Journal of Medical and Biological Research, 51(12), e7599.

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Oridonin Cytotoxicity Assays and Mechanism

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Oridonin was purchased from Sigma-Aldrich (St. Louis, MO, USA). For in vitro studies, Oridonin was dissolved in dimethyl sulfoxide (DMSO) to create a stock solution (0.1 mol/L), which was stored at −20°C. To prepare working solutions, the stock solution was further diluted with culture media to yield the desired Oridonin concentration. Control cells were treated with an equal volume of vehicle. The DMSO concentration was kept below 0.1% in cell culture and did not have any detectable effect on cell growth or cell death.
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT), Hoechst 33342, annexin V-FITC, propidium iodide (PI), and Rhodamine 123 were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Primary antibodies against caspase-3, caspase-9, NF-κB, Bax, Bcl-2, PARP-1, cytochrome c, β-actin, and secondary antibodies (goat-anti-rabbit or goat-anti-mouse) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against cyclin A, cyclin B1, and cyclin D1 were purchased from Epitomics (Burlingame, CA, USA).

Bao R., Shu Y., Wu X., Weng H., Ding Q., Cao Y., Li M., Mu J., Wu W., Ding Q., Tan Z., Liu T., Jiang L., Hu Y., Gu J, & Liu Y. (2014). Oridonin induces apoptosis and cell cycle arrest of gallbladder cancer cells via the mitochondrial pathway. BMC Cancer, 14, 217.

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High-Throughput Compound Screening for Drug Discovery

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Two collections of compounds assembled at the NCATS were screened A collection of 44,420 diverse small molecule drugs; and the Mechanism Interrogation PlatE 4.0 collection of 386 pharmacologically active small molecules (22 (link)). The active compounds were re-purchased for repeat and secondary assays. Milciclib (catalog #PHA-848125), MK-0752 (#HY-10974) and PP-121 (#HY-10372) were purchased from MedChem Express. Ouabain (#1076) and AMG-51 (#SYN-111) were purchased from Fisher Scientific. Vardenafil (#Y0001647) and Oridonin (#O9639) were purchased from Sigma-Aldrich. Peruvoside (# P227570) was purchased from Toronto Research Chemicals. NCGC00117362 (#D233–0871), NCGC00111761 (#C686–0165), NCGC00117328 (#D233–0834), NCGC00117505 (#D244–0327), NCGC00117477 (#D244–0252), NCGC00117166 (#D233–0497), NCGC00115018 (#D053–0260), NGC00117330 (#D233–0835) and NCGC00117364 (#D233–0885) were purchased from ChemDiv. Collagen type I (rat-tail), and fibronectin (human) were purchased from BD Biosciences. Rabbit anti-phospho m-TOR against Ser²⁴⁴⁸ (#2971), rabbit anti-mTOR (#2972), rabbit anti-phospho Rb against Ser^807/811 (#8516, clone D20B12), mouse anti-Rb (#9309), mouse anti-Cdk6 (#3136), lysis buffer (#9803), and 3x Blue Loading Buffer (#56036) were purchased from Cell Signaling Technology. Mouse anti-Cdk1/Cdk2 (#sc-53219) was purchased from Santa Cruz Biotechnology Inc.

Kenny H.A., Lal-Nag M., Shen M., Kara B., Nahotko D.A., Wroblewski K., Fazal S., Chen S., Chiang C.Y., Chen Y.J., Brimacombe K.R., Marugan J., Ferrer M, & Lengyel E. (2019). Quantitative high-throughput screening using an organotypic model identifies compounds that inhibit ovarian cancer metastasis. Molecular cancer therapeutics, 19(1), 52-62.

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Oridonin Mitigates Cisplatin-Induced Toxicity

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Male C57BL/6 mice were obtained from HyoSung Science (Daegu, Korea) and maintained at a temperature of at 20-24°C and humidity of 60-70%. The mice were grouped into three groups (n = 8 per group): the control group, the CP group, and the CP+Ori group. The CP group received a single intraperitoneal injection of cisplatin (20 mg/kg; Sigma-Aldrich, St. Louis, MO, USA). The CP+Ori group was given an intraperitoneal administration of oridonin (15 mg/kg; dissolved in DMSO; Sigma-Aldrich) daily for 3 consecutive days, starting from 1 hour after cisplatin injection. The control group received intraperitoneal injections of an equal volume of DMSO daily for 3 consecutive days. All mice were sacrificed 72 hours after a single dose of cisplatin. The doses of oridonin and cisplatin were selected based on the results of previous studies [10 (link), 13 (link)]. All animal procedures were approved by the Institutional Animal Care and Use Committee of the Daegu Catholic University Medical Center (DCIAFCR-200626-12-Y).

Gu H., Gwon M.G., Kim J.H., Leem J, & Lee S.J. (2022). Oridonin Attenuates Cisplatin-Induced Acute Kidney Injury via Inhibiting Oxidative Stress, Apoptosis, and Inflammation in Mice. BioMed Research International, 2022, 3002962.

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Oridonin-Loaded Nanoparticle Preparation

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ORI loaded NPs (ORI-NPs) were prepared using emulsification and evaporation method as previously described with modifications (Hua et al., 2017 (link)). Briefly, 20 mg of Oridonin (Sigma) and 60 mg of PEG/PLGA (PEG₅₀₀₀-PLGA_28,000, Sigma) were dissolved in 5 ml of dichloromethane as the oil (O) phase, while 20 ml of PVA (1%, w/w, Sigma) was used as external the water (W) phase. The oil phase was sonicated for 40 s at 100 Watt on ice to form the first emulsification, then it was dropped into W phase and sonicated for another 40 s to form the second emulsification. Finally, the nanoparticles were harvested by centrifuging at 12,000 rpm for 20 min and washed three times with water.
The size distribution and zeta potential of the nanoparticles was determined using a Zetasizer Nano ZS (Malvern Instruments, United Kingdom). The morphology of the as-prepared ORI-NPs was characterized by atomic force microscopy (AFM, Autoprobe CP Research, Veeco). AFM scanning were performed on Dimension 3100 system with a tapping mode.
Characterization of the products was confirmed by ¹H NMR spectrometer (Bruker AVANCE 400 NMR spectrometer, United States) with DMSO as the solvent and Fourier-transform infrared spectrometer (FTIR, Magna FTIR-750). FTIR spectra were obtained from a neat film cast from the chloroform copolymer solution between KBr tablets.

Zhao Y., Xiao W., Peng W., Huang Q., Wu K., Evans C.E., Liu X, & Jin H. (2021). Oridonin-Loaded Nanoparticles Inhibit Breast Cancer Progression Through Regulation of ROS-Related Nrf2 Signaling Pathway. Frontiers in Bioengineering and Biotechnology, 9, 600579.

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