| Targets |
Not applicable. 6alpha-Hydroxy-5alpha-cholestane is an oxysterol, an oxidized cholesterol derivative. It does not have a specific pharmacological target in the traditional sense, but rather functions as a signaling lipid that influences oxidative stress pathways. It promotes superoxide anion production, indicating it enhances the activity of cellular NADPH oxidases or mitochondrial electron transport chain complexes. Oxysterols also regulate cholesterol homeostasis via liver X receptors (LXRs) and other nuclear receptors.
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| ln Vitro |
6alpha-Hydroxy-5alpha-cholestane is an oxysterol that promotes the production of superoxide anions in SK-N-BE cells at concentrations of 50 uM and 100 uM. This demonstrates its ability to induce reactive oxygen species (ROS) formation in human neuroblastoma cells. The compound is classified as a member of the oxysterol family and is used in research on oxidative stress, neurotoxicity, and cholesterol metabolism. The specific mechanism of superoxide production is not detailed.
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| ln Vivo |
No specific in vivo data was found for 6alpha-Hydroxy-5alpha-cholestane as a drug. The compound is an oxysterol used for research into oxidative stress and neurodegenerative disease models. Oxysterols are known to accumulate in atherosclerotic plaques and in neurodegenerative conditions such as Alzheimer's disease. It is not a drug candidate; it is used as a research tool to study the role of oxysterol-induced oxidative stress in disease pathophysiology.
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| Enzyme Assay |
Not applicable. As a small-molecule lipid, 6alpha-Hydroxy-5alpha-cholestane can be tested in cell-free assays for its ability to generate superoxide anions in the presence of relevant enzymes such as NADPH oxidase. However, the primary activity data available was generated in cell-based assays. A typical cell-free assay for superoxide production would use purified NADPH oxidase and detect superoxide via cytochrome c reduction or lucigenin chemiluminescence.
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| Cell Assay |
Standard cell-based ROS detection protocol: SK-N-BE human neuroblastoma cells are seeded in 96-well black-walled clear-bottom plates. Cells are treated with 6alpha-Hydroxy-5alpha-cholestane at concentrations of 25, 50, 100, and 200 uM for 24-48 hours. After treatment, cells are washed with PBS and incubated with a ROS-sensitive fluorescent dye such as DCFH-DA (10 uM) or dihydroethidium (DHE) for 30-60 minutes at 37degC. Fluorescence intensity (Ex 485 nm/Em 530 nm for DCFH-DA; Ex 518 nm/Em 605 nm for DHE) is measured in a plate reader to quantify intracellular ROS levels. Cell viability is measured concurrently by MTT or CellTiter-Glo to ensure effects are not due to cytotoxicity.
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| Animal Protocol |
No specific in vivo protocol was found for 6alpha-Hydroxy-5alpha-cholestane. A general protocol for studying oxidative stress in a neurodegenerative disease model: Male C57BL/6 mice (8-12 weeks old) are administered 6alpha-Hydroxy-5alpha-cholestane intracerebroventricularly or systemically (e.g., 10-50 mg/kg, IP) daily for 7-14 days. Brain tissue is collected for measurement of superoxide dismutase (SOD) activity, malondialdehyde (MDA) levels (lipid peroxidation marker), and glutathione (GSH) content. Neuronal damage is assessed by histological staining (Nissl stain) and immunohistochemistry for markers of oxidative stress such as 4-HNE or 8-OHdG.
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| ADME/Pharmacokinetics |
No specific PK data was found. As an oxysterol, 6alpha-Hydroxy-5alpha-cholestane is lipophilic and would be expected to be distributed to lipid-rich tissues including the brain, liver, and adipose tissue. Oxysterols are typically metabolized by hydroxylation and sulfation and excreted in bile. Its half-life in plasma is likely short due to rapid hepatic clearance. No formal PK studies are described in the summarized literature.
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| Toxicity/Toxicokinetics |
No toxicity data was found for 6alpha-Hydroxy-5alpha-cholestane. As an oxysterol that promotes superoxide anion production, it may be toxic to cells at higher concentrations (≥50 uM in SK-N-BE cells), contributing to oxidative stress-induced cell damage and death. In the context of neurodegenerative diseases, oxysterols have been implicated as contributing factors to neuronal dysfunction and death. Standard chemical safety precautions apply.
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| References | |
| Additional Infomation |
6alpha-Hydroxy-5alpha-cholestane (5alpha-Cholestane-3beta,6alpha-diol) has molecular formula C27H48O2 and molecular weight 404.67. Purity reported as 95.8%. It is an oxysterol that promotes superoxide anion production in SK-N-BE neuroblastoma cells at 50-100 uM. It is a research tool for oxidative stress and neurodegenerative disease research. For research use only, not for human therapeutic applications.
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| CAS # |
41083-73-2
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|---|---|
| Related CAS # |
6α-Hydroxy-5α-cholestane-d7;1246302-83-9
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| Appearance |
Typically exists as solids at room temperature
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| LogP |
6.439
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| Synonyms |
5α-Cholestane-3β,6α-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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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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