| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| Targets |
RORα (RAR-related orphan receptor alpha, also known as NRIF1) - Neoruscogenin shows potent agonist activity with an EC50 of 0.11 μM in a pull-down assay measuring TIF2-BAP recruitment to GST-RORα. [1]
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| ln Vitro |
- Neoruscogenin significantly induced Gal4RE-luc reporter activity in COS7 cells expressing Gal4(DBD)-RORα protein (treatment with 10 μM Neoruscogenin), whereas reporter activity was only minimally induced in COS7 cells expressing the Gal4-DBD construct. [1]
- In HepG2 hepatoma cells, treatment with Neoruscogenin for 24 hours significantly induced the expression of well-established RORα target genes G6Pase and Bmal1, as measured by quantitative PCR. Expression was normalized to the 36B4 gene, and no toxicity was associated with the treatment. [1] |
| ln Vivo |
- In mice treated orally with Neoruscogenin at 3 mg/kg/day for 7 days, quantitative PCR analysis of liver samples showed significant upregulation of classical RORα target genes, including Bmal1, Cyp7b1, and G6Pase, with no change in the expression of the RORα receptor itself. Additionally, hepatic expression of Lpin2 and Angptl4, two genes that are significantly downregulated in RORα-deficient staggerer mice, was induced by Neoruscogenin treatment. [1]
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| Enzyme Assay |
A pull-down assay was performed to measure TIF2 protein recruitment on immobilized RORα protein. High-binding capacity streptavidin-coated plates were coated overnight with biotin-conjugated, affinity-purified anti-GST antibodies. After washing with TBSTween buffer (20 mM Tris-HCl pH 7.2, 150 mM NaCl, 0.05% Tween), the binding reaction was set up in binding buffer (15 mM Tris-HCl pH 8.0, 15 mM glycerol, 0.15 mM EDTA, 0.04% NP40, 22.5 mM KCl, 0.25 mg/mL fatty acid-free bovine serum albumin, 1 mM dithiothreitol, 1 mM benzamidine) with antiprotease tablets. Per reaction, 0.12 μg GST-RORα protein and 0.10 μg TIF2-BAP protein were used. Compounds were added as DMSO solutions. The binding reaction was performed overnight at 4°C. After washing, Lumi-Phos WB substrate was added (50 μL per well) and incubated for 30 min at 37°C. The luminescent signal was read with a microplate reader (integration time 500 ms). To verify specificity, hit compounds were tested in counterscreen assays including: (1) recruitment of a mutated TIF2-BAP protein (with LXXLL motifs mutated to LXXAA) on GST-RORα, (2) recruitment of TIF2-BAP on a GST protein fragment without ROR-LBD, and (3) effect of the compound on chemiluminescent signal produced by GST-BAP protein. Neoruscogenin was identified as a potent agonist with an EC50 of 0.11 μM in this assay. [1]
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| Cell Assay |
- COS7 cells were transfected with either Gal4-DBD or Gal4-RORα in addition to a Gal4-RE-luc reporter plasmid, and treated with either DMSO or 10 μM Neoruscogenin. Reporter activity (relative light units, RLU) was measured 24 hours upon stimulation. Neoruscogenin significantly induced Gal4RE-luc reporter activity only in cells expressing Gal4-RORα, not in cells expressing Gal4-DBD alone. [1]
- HepG2 hepatoma cells were treated with Neoruscogenin for 24 hours. RNA was harvested, and RORα target gene (G6Pase, Bmal1) expression was studied by quantitative PCR. The expression of target genes was normalized to the expression of the 36B4 gene. Treatment was not associated with toxicity as assessed by an unspecified assay. [1] |
| Animal Protocol |
- For pharmacodynamic studies: Mice were treated with Neoruscogenin at 3 mg/kg/day per os for 7 days. After treatment, liver samples were collected, RNA was extracted, and gene expression was measured by quantitative PCR. [1]
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| ADME/Pharmacokinetics |
- Neoruscogenin showed excellent metabolic stability: 88% of the parent compound remained intact upon 1-hour incubation with mouse primary hepatic microsome preparations, and 73% remained intact with human primary hepatic microsome preparations. [1]
- In classic pharmacokinetic studies in mice after a single oral dose of 10 mg/kg, circulating Neoruscogenin was detected in plasma with a Cmax of 1.1 μmol/L, a Tmax of 3 hours, and an AUC0-24h of 3561 ng/mL·h. Significantly higher exposure was detected in brain samples. The brain tissue exposure (Cmax) was estimated to be 150-fold above the EC50. [1] |
| Toxicity/Toxicokinetics |
- In HepG2 cells, treatment with Neoruscogenin was not associated with toxicity as determined by an unspecified assay (likely a cell viability assay, e.g., from Promega). [1]
- Neoruscogenin did not significantly activate most steroid or xenobiotic nuclear receptors, with the exception of a modest PXR activation (1.8-fold reporter induction in cells expressing GAL4(DBD)-PXR vs. 1.3-fold induction in cells without GAL4(DBD)-PXR). This induction was very modest compared to classic PXR reference ligands such as rifampicin (4- to 5-fold increase). [1] |
| References |
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| Additional Infomation |
Neoruscogenin is a triterpenoid compound. It has been reported that Heliotropium indicum contains Neoruscogenin, and relevant data are available for reference.
- Neoruscogenin is a close analogue of (25S)-ruscogenin, which was originally isolated from Dracaena cambodiana (Agavaceae). (25S)-ruscogenin showed an EC50 of 0.78 μM in the pull-down assay, while Neoruscogenin was more potent with an EC50 of 0.11 μM. [1] - Neoruscogenin is commercially available in gram quantities, making it a practical pharmacological probe for in vitro and in vivo validation studies of RORα biology. [1] - The study describes a high-throughput screening-compatible procedure using fractionated plant extracts to identify naturally occurring RORα ligands, with Neoruscogenin emerging as a potent and bioavailable agonist. [1] |
| Molecular Formula |
C27H40O4
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|---|---|
| Molecular Weight |
428.6041
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| Exact Mass |
428.292
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| CAS # |
17676-33-4
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| PubChem CID |
9910474
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| Appearance |
White to off-white solid
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
566.6±50.0 °C at 760 mmHg
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| Melting Point |
196-198℃
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| Flash Point |
296.4±30.1 °C
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| Vapour Pressure |
0.0±3.5 mmHg at 25°C
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| Index of Refraction |
1.585
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| LogP |
4.21
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
31
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| Complexity |
820
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| Defined Atom Stereocenter Count |
11
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| SMILES |
O1[C@]2(C([H])([H])C([H])([H])C(=C([H])[H])C([H])([H])O2)[C@@]([H])(C([H])([H])[H])[C@@]2([H])[C@]1([H])C([H])([H])[C@@]1([H])[C@]3([H])C([H])([H])C([H])=C4C([H])([H])[C@]([H])(C([H])([H])[C@]([H])([C@]4(C([H])([H])[H])[C@@]3([H])C([H])([H])C([H])([H])[C@@]12C([H])([H])[H])O[H])O[H]
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| InChi Key |
ALTRINCJVPIQNK-NHIXJPGBSA-N
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| InChi Code |
InChI=1S/C27H40O4/c1-15-7-10-27(30-14-15)16(2)24-22(31-27)13-21-19-6-5-17-11-18(28)12-23(29)26(17,4)20(19)8-9-25(21,24)3/h5,16,18-24,28-29H,1,6-14H2,2-4H3/t16-,18+,19+,20-,21-,22-,23+,24-,25-,26-,27+/m0/s1
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| Chemical Name |
(1S,2S,4S,6R,7S,8R,9S,12S,13R,14R,16R)-7,9,13-trimethyl-5'-methylidenespiro[5-oxapentacyclo[10.8.0.02,9.04,8.013,18]icos-18-ene-6,2'-oxane]-14,16-diol
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~116.66 mM)
Ethanol : ~10 mg/mL (~23.33 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (5.83 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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. Solubility in Formulation 2: ≥ 0.56 mg/mL (1.31 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 5.6 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 0.56 mg/mL (1.31 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3332 mL | 11.6659 mL | 23.3318 mL | |
| 5 mM | 0.4666 mL | 2.3332 mL | 4.6664 mL | |
| 10 mM | 0.2333 mL | 1.1666 mL | 2.3332 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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