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| 5mg |
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| Targets |
beta-Sitosterol-d7 is a stable isotope-labeled internal standard that mimics the biological activity of its non-labeled counterpart, beta-Sitosterol. beta-Sitosterol is a plant sterol with a chemical structure very similar to cholesterol. Its primary mechanism of action involves competitively inhibiting the absorption of both dietary and biliary cholesterol in the small intestine. It does this by co-crystallizing with cholesterol in the intestinal lumen and displacing cholesterol from mixed micelles, thereby reducing the amount of cholesterol available for absorption. Systemically, beta-Sitosterol has been shown to inhibit the activity of 5alpha-reductase, the enzyme responsible for converting testosterone to the more potent androgen dihydrotestosterone (DHT). This activity is thought to underlie its clinical use in treating benign prostatic hyperplasia (BPH).
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1].
The in vitro biological activity of beta-Sitosterol-d7 is presumed to be identical to its non-labeled parent, beta-Sitosterol. In vitro studies using human prostate stromal cells (WPMY-1) have shown that beta-Sitosterol (10-50 uM) significantly inhibits cell proliferation and induces apoptosis. Mechanistically, these effects are mediated by the activation of the caspase-3 apoptotic pathway and the downregulation of pro-inflammatory cytokines such as IL-6 and TNF-alpha. In Caco-2 human intestinal epithelial cells, beta-Sitosterol has been shown to reduce cholesterol absorption by downregulating the expression of the Niemann-Pick C1-Like 1 (NPC1L1) protein, a key cholesterol transporter. Additionally, it exhibits anti-inflammatory properties by suppressing the NF-kappaB signaling pathway in LPS-stimulated macrophages. beta-Sitosterol-d7 is used as an LC-MS internal standard to accurately quantify beta-sitosterol levels in these in vitro systems. |
| ln Vivo |
beta-Sitosterol-d7 is a stable isotope-labeled compound, and the in vivo activity of its non-labeled counterpart is well-established. In animal models, dietary administration of beta-Sitosterol (0.5-2% of diet) to hypercholesterolemic rodents leads to a significant reduction (15-30%) in plasma total cholesterol and LDL-cholesterol levels by inhibiting intestinal cholesterol absorption. In the prostate, in vivo studies using rats with testosterone-induced benign prostatic hyperplasia (BPH) have shown that oral administration of beta-Sitosterol (20-50 mg/kg) reduces prostate weight and diminishes the histological signs of hyperplasia. This effect is linked to the downregulation of 5alpha-reductase enzyme expression and activity. The labeled compound (beta-Sitosterol-d7) is used as an internal standard for the accurate quantification of beta-sitosterol levels in plasma, tissue, and fecal samples in these animal models.
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| Enzyme Assay |
A generic non-cell-based assay for beta-Sitosterol-d7 is a cholesterol micelle solubility assay to assess its capacity to displace cholesterol. First, prepare a simulated intestinal fluid (SIF) buffer (pH 6.5). Prepare a stock micellar solution containing 5 mM cholesterol, 5 mM beta-sitosterol (unlabeled), 10 mM oleic acid, 10 mM monoolein, and 10 mM sodium taurocholate in SIF buffer. Sonicate the mixture to achieve a clear solution. For the assay, add increasing concentrations of beta-sitosterol (0.1-10 mM) to a fixed concentration of cholesterol (5 mM) in the micellar solution. Incubate the mixture at 37degC for 24 hours with gentle shaking. After incubation, filter the micellar solution through a 0.22 um filter to remove any precipitated crystals. Quantify the cholesterol concentration remaining in the filtrate using an enzymatic colorimetric assay or HPLC. Calculate the concentration of beta-sitosterol required to reduce cholesterol solubility by 50% (EC₅0). Use beta-Sitosterol-d7 as an internal standard in LC-MS to validate the concentration of beta-sitosterol in the test samples.
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| Cell Assay |
A standard in vitro cell-based assay for beta-Sitosterol-d7 involves studying its anti-proliferative effects on prostate cancer cells. Culture LNCaP or PC-3 human prostate cancer cells in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37degC in a 5% CO2 incubator. Seed cells at 5,000 cells per well in 96-well plates and allow them to attach overnight. Treat the cells with increasing concentrations of unlabeled beta-sitosterol (10, 25, 50, 100 uM) or vehicle control (ethanol) for 24, 48, and 72 hours. After the treatment period, add 10 uL of CCK-8 solution to each well and incubate for 2 hours. Measure the absorbance at 450 nm using a microplate reader. Calculate the half-maximal inhibitory concentration (IC₅0) for cell growth inhibition from the dose-response curve. Use beta-Sitosterol-d7 as an internal standard in an LC-MS analysis of the culture medium to precisely determine the concentration of beta-sitosterol that the cells were exposed to during the course of the experiment.
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| Animal Protocol |
A typical in vivo protocol for beta-Sitosterol-d7 uses a rat model of benign prostatic hyperplasia (BPH). Use male Wistar rats (200-250 g). Induce BPH by daily subcutaneous injections of testosterone propionate (5 mg/kg) for 28 days. Randomize the rats into groups (n=8-10 per group): sham (vehicle only), BPH control (testosterone + vehicle), and treatment groups (testosterone + unlabeled beta-Sitosterol at 10, 20, or 50 mg/kg body weight, administered orally once daily). After 4 weeks, sacrifice the animals and weigh the ventral prostate. Fix one lobe of the prostate in formalin for histological analysis (H&E staining). Homogenize the remaining prostate tissue. Measure the activity of 5alpha-reductase in the tissue homogenate using a commercially available ELISA kit. Use beta-Sitosterol-d7 as the internal standard in an LC-MS analysis of plasma and prostate homogenates to accurately quantify the concentrations of beta-sitosterol and correlate them with the degree of BPH reduction.
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| ADME/Pharmacokinetics |
beta-Sitosterol-d7 is an isotopically labeled internal standard and thus its PK is the same as its non-labeled parent. beta-Sitosterol is poorly absorbed from the gastrointestinal tract, with an oral bioavailability of less than 5% in humans. This is due to its low solubility in the intestinal lumen and its rapid efflux back into the lumen by ATP-binding cassette (ABC) transporters such as ABCG5 and ABCG8. Following absorption, it is extensively distributed in the body, with highest concentrations found in the liver. It undergoes enterohepatic circulation. The elimination half-life of beta-sitosterol is approximately 24 hours in humans. It is primarily excreted unchanged in the feces, with only trace amounts excreted in the urine. The labeled beta-Sitosterol-d7 serves as a critical internal standard in PK studies, allowing researchers to accurately measure plasma and tissue concentrations of phytosterols using mass spectrometry, thereby obtaining more reliable estimates of these parameters.
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| Toxicity/Toxicokinetics |
beta-Sitosterol-d7 itself is not intended for human use, so its toxicity is inferred from the unlabeled beta-sitosterol, which is generally recognized as safe (GRAS) by the FDA. beta-Sitosterol is a dietary phytosterol found in many foods, and it is widely used as an over-the-counter supplement for lowering cholesterol and treating benign prostatic hyperplasia (BPH). No significant adverse effects have been reported at typical supplement doses (up to 3 grams per day). Mild gastrointestinal symptoms such as bloating or constipation may occur in some individuals. There is a rare genetic condition, sitosterolemia, in which individuals hyperabsorb phytosterols, leading to premature coronary artery disease. For individuals without this condition, beta-sitosterol is considered safe. Standard laboratory precautions should be taken when handling the labeled compound for research purposes.
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| References | |
| Additional Infomation |
beta-Sitosterol-d7 (Mixture of Diastereomers) is a stable isotope-labeled (deuterated) version of beta-Sitosterol, a common plant sterol. It is used as an internal standard for the accurate quantification of beta-sitosterol in complex matrices like food, plasma, and tissue samples by GC-MS or LC-MS. beta-Sitosterol itself has multiple established pharmacological activities. It is a cholesterol-lowering agent that reduces intestinal cholesterol absorption. Clinically, it is used to manage hypercholesterolemia. It is also approved in Europe for the treatment of benign prostatic hyperplasia (BPH) due to its ability to inhibit 5alpha-reductase and reduce DHT levels. Research continues to explore its anti-cancer, anti-inflammatory, and immunomodulatory properties. This mixture of diastereomers is provided for research purposes only and not for human use.
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| Molecular Formula |
C29H50O
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| Molecular Weight |
421.75
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| Exact Mass |
421.43
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| CAS # |
2260669-28-9
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| PubChem CID |
57347730
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| Appearance |
White to off-white solid powder
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| LogP |
9.3
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
1
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
30
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| Complexity |
634
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| Defined Atom Stereocenter Count |
8
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| SMILES |
[C@H]1(CC2=CC[C@]3([H])[C@@]([H])([C@]2(CC1)C)CC[C@]1([C@@]3([H])CC[C@]1([H])[C@@H](CCC(C(C([2H])([2H])[2H])([2H])C([2H])([2H])[2H])CC)C)C)O
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| InChi Key |
KZJWDPNRJALLNS-GNYYFJBRSA-N
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| InChi Code |
InChI=1S/C29H50O/c1-7-21(19(2)3)9-8-20(4)25-12-13-26-24-11-10-22-18-23(30)14-16-28(22,5)27(24)15-17-29(25,26)6/h10,19-21,23-27,30H,7-9,11-18H2,1-6H3/t20-,21?,23+,24+,25-,26+,27+,28+,29-/m1/s1/i2D3,3D3,19D
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| Chemical Name |
(3S,8S,9S,10R,13R,14S,17R)-10,13-dimethyl-17-[(2R)-6,7,7,7-tetradeuterio-5-ethyl-6-(trideuteriomethyl)heptan-2-yl]-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol
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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) |
THF: 50 mg/mL (118.55 mM)
Ethanol: 16.67 mg/mL (39.53 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (2.37 mM) (saturation unknown) in 10% EtOH + 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 10.0 mg/mL clear EtOH + stock solution to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3711 mL | 11.8554 mL | 23.7107 mL | |
| 5 mM | 0.4742 mL | 2.3711 mL | 4.7421 mL | |
| 10 mM | 0.2371 mL | 1.1855 mL | 2.3711 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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