| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
| Targets |
Obeticholic Acid-d4 targets the farnesoid X receptor (FXR, NR1H4), a nuclear receptor that is primarily expressed in the liver, intestine, kidney, and adrenal glands. FXR is a master regulator of bile acid, lipid, and glucose homeostasis. Upon binding to FXR, Obeticholic Acid activates the receptor, leading to its translocation to the nucleus, heterodimerization with retinoid X receptor (RXR), and binding to FXR response elements (FXREs) in the promoter regions of target genes. This results in the repression of bile acid synthesis (via SHP-dependent inhibition of CYP7A1) and the induction of bile acid export (via BSEP and other transporters). The parent compound has an EC50 of 99 nM for FXR activation. Obeticholic acid also has anti-inflammatory and anti-cholestatic effects.
|
|---|---|
| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers for quantification throughout the drug development process. Due to its potential to alter the pharmacokinetic and metabolic characteristics of medications, deuteration has drawn attention[1].
No specific in vitro activity data is available for the deuterated form. The parent compound Obeticholic Acid has been extensively characterized. In cell-free FXR binding assays, Obeticholic Acid binds to FXR with an EC50 of 99 nM for activation, as measured in a cell-based reporter gene assay using a luciferase reporter under the control of an FXR response element. It is significantly more potent than the endogenous bile acid chenodeoxycholic acid (CDCA, EC50 ∼ 30-50 microM). In human hepatoma Huh-7 cells, Obeticholic Acid (0.1-10 microM) induces the expression of FXR target genes, including small heterodimer partner (SHP) and bile salt export pump (BSEP), while repressing CYP7A1 and CYP8B1. It also reduces the expression of pro-inflammatory genes (TNF-alpha, IL-1beta, IL-6) in LPS-stimulated macrophages. |
| ln Vivo |
No specific in vivo data is provided for the deuterated form. The parent compound Obeticholic Acid is an FDA-approved drug for the treatment of primary biliary cholangitis (PBC). In preclinical studies, oral administration of Obeticholic Acid (1-30 mg/kg/day) in rodent models of cholestasis (e.g., bile duct ligation, ANIT-induced cholestasis) reduces serum bile acid levels, alkaline phosphatase (ALP), and bilirubin, and improves liver histology. In a mouse model of non-alcoholic steatohepatitis (NASH), Obeticholic Acid reduces hepatic steatosis, inflammation, and fibrosis. The clinical efficacy of Obeticholic Acid for PBC is well-established, with the compound producing significant reductions in ALP, a biomarker of cholestasis. The labeled internal standard is used for quantifying the parent drug in clinical PK studies.
|
| Enzyme Assay |
Obeticholic Acid-d4 is not used in cell-free assays for target engagement. It is used as an internal standard in analytical chemistry. For LC-MS/MS method validation, calibration standards are prepared by spiking known concentrations of non-labeled Obeticholic Acid into blank biological matrix (e.g., human plasma, rat plasma, or liver homogenate). A fixed concentration of Obeticholic Acid-d4 (e.g., 5-50 ng/mL) is added to each calibrator, QC sample, and study sample. Samples are prepared by protein precipitation with acetonitrile containing the internal standard or by solid-phase extraction (SPE). The supernatant or eluate is injected onto a reversed-phase C18 HPLC column coupled to a triple quadrupole mass spectrometer. Detection is performed in negative ion electrospray mode (ESI-). MRM transitions: parent drug m/z 417.3 → m/z 399.3; internal standard m/z 421.3 → m/z 403.3 (for d4). The peak area ratio is used for quantification.
|
| Cell Assay |
Obeticholic Acid-d4 is not typically used in cellular assays. For studies of the parent compound, a cell-based FXR reporter gene assay is used to measure FXR activation. Huh-7 cells are transiently transfected with an FXR expression plasmid and a luciferase reporter plasmid containing an FXR response element (FXRE). After 24 hours, cells are treated with varying concentrations of Obeticholic Acid (0.01 nM-10 microM) for 16-24 hours. Luciferase activity is measured using a standard luciferase assay kit, and the EC50 is calculated. For gene expression studies, Huh-7 cells or primary human hepatocytes are treated with Obeticholic Acid (0.1-10 microM) for 8-24 hours, and RNA is extracted for qRT-PCR analysis of FXR target genes (SHP, BSEP, CYP7A1). Alternatively, the compound is used to study FXR-mediated anti-inflammatory effects in LPS-stimulated RAW 264.7 macrophages. The deuterated internal standard is not used in these experiments.
|
| Animal Protocol |
Obeticholic Acid-d4 is not typically administered in vivo as a test compound. For pharmacokinetic studies, the non-deuterated Obeticholic Acid is administered to animals or human subjects. In a rat PK study, male Sprague-Dawley rats (200-250 g) receive a single oral dose of Obeticholic Acid (1-30 mg/kg) suspended in 0.5% methylcellulose or other oral vehicle. Blood samples are collected via tail vein at pre-dose, 0.25, 0.5, 1, 2, 4, 8, 12, 24, 36, and 48 hours post-dose. Plasma is separated by centrifugation. Obeticholic Acid concentrations are quantified by LC-MS/MS using Obeticholic Acid-d4 as internal standard. PK parameters (Cmax, Tmax, AUC, t½, oral bioavailability) are calculated using non-compartmental analysis. For tissue distribution studies, animals are euthanized at selected time points, and tissues (liver, kidney, intestine, brain) are collected, homogenized, and processed for LC-MS/MS analysis using the deuterated internal standard.
|
| ADME/Pharmacokinetics |
Obeticholic Acid-d4 (free base) has a molecular weight of approximately 424.67 g/mol (depending on the degree of deuteration). The compound is supplied as a solid and should be stored at -20degC in a sealed container, protected from light and moisture, where it is stable for up to 3 years. For solution storage, stock solutions in DMSO or methanol should be stored at -80degC for up to 6 months. The compound is soluble in DMSO (10-20 mg/mL) and methanol. For analytical use, working solutions are prepared by dilution in acetonitrile/water (50:50) or mobile phase. The internal standard is typically added to samples at a concentration of 5-50 ng/mL. The product is for research use only, not for human use.
|
| Toxicity/Toxicokinetics |
This product is for research use only and is not for human therapeutic use. No specific toxicity data is available for the deuterated form. The parent compound Obeticholic Acid is an FDA-approved drug with an established safety profile. Common adverse effects in patients with PBC include pruritus (itching), fatigue, abdominal pain, and constipation. Higher doses are associated with increased pruritus. The deuterated internal standard is used at minute quantities (ng/mL levels in analytical solutions) that pose negligible risk. Standard laboratory safety practices (gloves, lab coat, safety glasses) should be followed. Avoid inhalation of powder. The compound is intended for use as an internal standard only.
|
| References |
[1]. Russak EM, et al. Impact of Deuterium Substitution on the Pharmacokinetics of Pharmaceuticals. Ann Pharmacother. 2019;53(2):211-216.
[2]. Pellicciari R, et al. 6alpha-ethyl-chenodeoxycholic acid (6-ECDCA), a potent and selective FXR agonist endowed with anticholestatic activity. J Med Chem. 2002 Aug 15;45(17):3569-72. [3]. Fiorucci S, et al. Protective effects of 6-ethyl chenodeoxycholic acid, a farnesoid X receptor ligand, in estrogen-induced cholestasis. J Pharmacol Exp Ther. 2005 May;313(2):604-12. [4]. Ghebremariam YT, et al. FXR agonist INT-747 upregulates DDAH expression and enhances sensitivity in high-salt fed Dahl rats. PLoS One. 2013 Apr 4;8(4):e60653. [5]. Verbeke L, et al. The FXR Agonist Obeticholic Acid Prevents Gut Barrier Dysfunction and Bacterial Translocation in Cholestatic Rats. Am J Pathol. 2015 Feb;185(2):409-19. |
| Additional Infomation |
Obeticholic Acid (brand name Ocaliva) was approved by the FDA in 2016 for the treatment of primary biliary cholangitis (PBC) in combination with ursodeoxycholic acid (UDCA) in adults who have an inadequate response to UDCA, or as monotherapy. It is the first FXR agonist approved for clinical use. Obeticholic Acid-d4 is a stable isotopic internal standard used for accurate quantification by mass spectrometry. The incorporation of four deuterium atoms provides a mass shift of +4 Da relative to the parent drug. This internal standard is essential for correcting matrix effects and extraction recovery in bioanalytical assays. The CAS number for the non-deuterated drug is 459789-99-2. This product is not a drug; it is a research chemical for analytical use. Supplier information must not be included.
|
| Molecular Formula |
C25H38D4O4
|
|---|---|
| Related CAS # |
Obeticholic acid;459789-99-2
|
| Appearance |
Typically exists as solid at room temperature
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
