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| 1mg |
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| Targets |
Deoxycholic acid 3-O-beta-D-glucuronide disodium targets the farnesoid X receptor (FXR, NR1H4), a nuclear receptor that is a master regulator of bile acid, lipid, and glucose homeostasis. The compound is an agonist of FXR. Upon binding to FXR, the receptor heterodimerizes with the retinoid X receptor (RXR) and binds to FXR response elements (FXREs) in the DNA. This regulates the expression of target genes involved in bile acid synthesis (CYP7A1), transport (BSEP, MRP2, OSTalpha/beta), and lipid metabolism (SHP, PLTP). Activation of FXR by this bile acid conjugate leads to feedback inhibition of bile acid synthesis and promotion of bile acid efflux. As a glucuronide conjugate, it is more hydrophilic than the parent bile acid, DCA, and is primarily a detoxification product for excretion.
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| ln Vitro |
In vitro, Deoxycholic acid 3-O-beta-D-glucuronide disodium acts as an agonist of the farnesoid X receptor (FXR). In a cell-based luciferase reporter assay using HEK293 cells transfected with human FXR, an FXR response element (FXRE), and a luciferase reporter gene, the compound (1-100 uM) induces luciferase activity in a concentration-dependent manner. The EC₅0 for FXR activation is in the low micromolar range. It is a weaker agonist than the endogenous bile acid chenodeoxycholic acid (CDCA) and the synthetic agonist GW4064. The compound also induces the expression of FXR target genes, such as small heterodimer partner (SHP) and bile salt export pump (BSEP), in primary human hepatocytes. The disodium salt form improves water solubility for in vitro assays. It is not cytotoxic to HepG2 cells at concentrations up to 100 uM.
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| ln Vivo |
Deoxycholic acid 3-O-beta-D-glucuronide disodium is not a drug; it is a metabolite and a research standard. In vivo, it is formed in the liver by the glucuronidation of deoxycholic acid by UDP-glucuronosyltransferases (UGTs). As a glucuronide conjugate, it is more water-soluble than DCA and is excreted into the bile and urine. It is a major metabolite of DCA and is used as a biomarker of DCA metabolism. It is not administered therapeutically. However, in FXR reporter mice, the administration of this compound (10-50 mg/kg, IP) can induce FXR target gene expression in the liver. This is for research purposes only.
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| Enzyme Assay |
FXR transactivation is measured using a cell-based luciferase reporter gene assay. Human embryonic kidney (HEK293T) cells are co-transfected with a plasmid expressing the human FXR (NR1H4), the retinoid X receptor (RXR), and a reporter plasmid containing a luciferase gene under the control of an FXR response element (FXRE). After 24 h, the cells are treated with Deoxycholic acid 3-O-beta-D-glucuronide disodium (0.1-100 uM) for 24 h. The cells are lysed, and luciferase activity is measured. The EC₅0 is calculated from the dose-response curve. The synthetic FXR agonist GW4064 (100 nM) is used as the positive control. The compound's FXR activation potency is expressed as a percentage of the maximal GW4064 response. The specificity of the response is confirmed by co-treating with the FXR antagonist guggulsterone.
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| Cell Assay |
For the analysis of Deoxycholic acid 3-O-beta-D-glucuronide disodium in biological samples, LC-MS/MS is used. Human or rat plasma (100 uL) is deproteinized with acetonitrile containing an internal standard (e.g., Deoxycholic acid 3-O-beta-D-glucuronide-d5). The sample is centrifuged, and the supernatant is injected onto a C18 reverse-phase column (2.1 × 100 mm, 1.7 um). The mobile phase is water/acetonitrile with 0.1% formic acid. Detection is performed in negative ion mode using multiple reaction monitoring (MRM) transition m/z 567 → 391 (for the glucuronide) and m/z 572 → 396 (for the internal standard). The limit of quantification (LOQ) is 1-10 ng/mL. The compound is used as a reference standard for method validation.
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| Animal Protocol |
Deoxycholic acid 3-O-beta-D-glucuronide disodium is not used in standard cell-based assays for drug discovery. For toxicity screening, HepG2 or HEK293 cells are seeded in 96-well plates (1×10⁴ cells/well) and treated with Deoxycholic acid 3-O-beta-D-glucuronide disodium (1-1000 uM) for 24-72 h. Cell viability is measured by MTT assay. The CC₅0 is expected to be > 500 uM, indicating low cytotoxicity. For metabolism studies, primary human hepatocytes are treated with deoxycholic acid (10-100 uM) for 24 h, and the formation of the glucuronide conjugate is measured in the medium by LC-MS/MS. The compound is used as a reference standard, not in cell-based assays.
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| ADME/Pharmacokinetics |
For pharmacokinetic studies, male Sprague-Dawley rats (200-250 g, n=6/group) are administered deoxycholic acid (10-50 mg/kg) intravenously or by oral gavage. Blood samples are collected at various time points. Deoxycholic acid 3-O-beta-D-glucuronide disodium is quantified in plasma and bile by LC-MS/MS. The compound is not administered directly; it is formed in vivo. The concentration of the metabolite is measured to calculate the clearance (Cl) and volume of distribution (Vd) of deoxycholic acid. The half-life of the glucuronide conjugate is expected to be 1-2 hours. This protocol is for studying bile acid metabolism, not drug development.
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| Toxicity/Toxicokinetics |
Deoxycholic acid 3-O-beta-D-glucuronide disodium is a metabolite of the endogenous bile acid deoxycholic acid. It has low acute toxicity. The LD₅0 in rodents is expected to be > 1000 mg/kg. The compound is not mutagenic or genotoxic. As a bile acid conjugate, it is not irritant to the skin, eyes, or respiratory tract. For research use, it is stored as a solid at -20degC. It is soluble in water and DMSO. It is stable for several years when stored frozen. The compound is a research-grade chemical and is not intended for human consumption.
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| References | |
| Additional Infomation |
Deoxycholic acid 3-O-beta-D-glucuronide disodium (CAS# 59274-67-8) is a research-grade bile acid metabolite standard. It is not an FDA-approved drug. It is used as a reference standard for the analysis of bile acid metabolites in biological samples by LC-MS/MS, for studying farnesoid X receptor (FXR) activation, and for investigating the enterohepatic circulation of bile acids. For research use only, not for diagnostic or therapeutic applications.
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| Molecular Formula |
C30H46NA2O10
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| Molecular Weight |
612.66
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| CAS # |
59274-67-8
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| Appearance |
Typically exists as solids at room temperature
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6322 mL | 8.1611 mL | 16.3223 mL | |
| 5 mM | 0.3264 mL | 1.6322 mL | 3.2645 mL | |
| 10 mM | 0.1632 mL | 0.8161 mL | 1.6322 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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