| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
| Targets |
9-cis-Retinol acetate-d5 is a stable isotope-labeled internal standard. Its unlabeled parent, 9-cis-Retinol acetate, is a retinal analog. Retinal analogs exert their effects by binding to nuclear retinoic acid receptors (RARs) and retinoid X receptors (RXRs). RARs and RXRs are ligand-activated transcription factors that regulate the expression of genes involved in cellular differentiation, proliferation, and apoptosis. Specifically, 9-cis-retinoic acid (which is likely the active metabolite of 9-cis-Retinol acetate) is a high-affinity ligand for both RARs and RXRs, while all-trans-retinoic acid only binds to RARs. By binding to these receptors, the compound can modulate the development and function of the visual system and can influence cell cycle control. The labeled version is used to track these interactions analytically.
|
|---|---|
| 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].
9-cis-Retinol acetate-d5 is not directly used in cell-based assays; rather, its unlabeled form is. The unlabeled compound is a retinoid with significant biological activity. In vitro, 9-cis-Retinol acetate can be converted to 9-cis-retinoic acid, which is a potent activator of both retinoic acid receptors (RARs) and retinoid X receptors (RXRs). In reporter gene assays using cells transfected with RAR or RXR reporter plasmids, 9-cis-retinoic acid activates transcription with EC₅0 values in the low nanomolar range (1-10 nM). In human cancer cell lines (e.g., breast cancer MCF-7, prostate cancer LNCaP), treatment with 9-cis-retinoic acid (0.1-10 uM) induces cell cycle arrest and apoptosis by upregulating pro-apoptotic genes and downregulating anti-apoptotic genes. It also inhibits the proliferation of retinal pigment epithelial (RPE) cells. The labeled version is used as an internal standard to quantify the active retinoids in the cell culture medium. |
| ln Vivo |
9-cis-Retinol acetate-d5 is an analytical internal standard. The in vivo activity of its unlabeled parent compound is an area of active research. 9-cis-Retinol acetate is a prodrug that is converted to 9-cis-retinoic acid in the body. In animal models of retinal degeneration (e.g., Rpe65-/- mice, a model of Leber congenital amaurosis), oral or intraperitoneal administration of 9-cis-Retinol acetate has been shown to restore visual function. This is because it acts as a visual cycle modulator, bypassing the defective RPE65 enzyme to regenerate 11-cis-retinal, the chromophore essential for vision. In cancer models, such as in mice with xenografted human breast tumors, administration of 9-cis-retinoic acid (5-20 mg/kg) has been shown to inhibit tumor growth and metastasis. The compound also reduces the severity of acute promyelocytic leukemia (APL) by inducing differentiation of the malignant promyelocytes. 9-cis-Retinol acetate-d5 is used to quantify these active species.
|
| Enzyme Assay |
A generic non-cell-based assay for 9-cis-Retinol acetate-d5 involves its use as an internal standard in an LC-MS/MS method for retinoid analysis. Prepare standard stock solutions of unlabeled 9-cis-Retinol acetate in ethanol (1 mg/mL), protected from light. Prepare a separate stock solution of the internal standard (9-cis-Retinol acetate-d5) at the same concentration. Prepare calibration standards by spiking the unlabeled analyte into a blank matrix (e.g., charcoal-stripped human serum) to achieve concentrations ranging from 1 ng/mL to 1000 ng/mL. Add a fixed concentration of the internal standard (e.g., 50 ng/mL) to each calibration standard. Also prepare blank and double-blank samples. For sample preparation, perform liquid-liquid extraction with hexane. Evaporate the organic layer under nitrogen and reconstitute in methanol. Use a C18 reverse-phase column for LC separation. Analyze by LC-MS/MS in positive ion mode. Monitor the mass transitions: m/z 329.2 → 255.2 for 9-cis-Retinol acetate, and m/z 334.2 → 260.2 for the d5 internal standard. Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration. All procedures must be performed under amber or subdued light to prevent photodegradation of retinoids.
|
| Cell Assay |
A standard in vitro cell-based protocol for the unlabeled 9-cis-Retinol acetate is used to study gene expression regulation. Culture MCF-7 human breast cancer cells in DMEM supplemented with 10% charcoal-stripped fetal bovine serum (to remove endogenous retinoids) and 1% penicillin/streptomycin at 37degC in a 5% CO2 incubator. Seed cells in 6-well plates at 2×10⁵ cells/well and allow them to attach for 24 hours. Treat the cells with increasing concentrations (1 nM, 10 nM, 100 nM, 1 uM) of unlabeled 9-cis-Retinol acetate or vehicle control (ethanol, final concentration <0.1%) for 24-48 hours. After treatment, collect the cells and extract total RNA using a kit. Perform quantitative real-time PCR (qRT-PCR) to measure the expression levels of retinoid-responsive genes such as RARbeta, CYP26A1, and CRABPII. For validation, collect the culture media, add 9-cis-Retinol acetate-d5 as the internal standard, and analyze by LC-MS/MS to quantify the concentrations of the parent compound and its active metabolites (e.g., 9-cis-retinoic acid) to which the cells were exposed.
|
| Animal Protocol |
A standard in vivo protocol for 9-cis-Retinol acetate-d5 involves a study in a mouse model of retinal degeneration (e.g., Rpe65 knockout mice). These mice have a defect in the visual cycle leading to blindness. Use 4-week-old Rpe65-/- mice. Administer unlabeled 9-cis-Retinol acetate via intraperitoneal injection (5-20 mg/kg) or oral gavage daily for 7-14 days. Include a vehicle control group and a wild-type control group. Assess retinal function before and after treatment using electroretinography (ERG) to measure scotopic and photopic responses. After the final dose, euthanize the mice and collect blood, liver, and eyes. For bioanalysis, homogenize the eyes (neural retina and RPE) in PBS. Perform a lipid extraction to isolate retinoids. Add 9-cis-Retinol acetate-d5 as the internal standard to all samples (blood, liver, eye) before extraction. Analyze by LC-MS/MS to quantify the levels of the administered prodrug and the generated 11-cis-retinal and 9-cis-retinoic acid. This allows for correlation of drug levels with restored visual function.
|
| ADME/Pharmacokinetics |
The pharmacokinetics (PK) of 9-cis-Retinol acetate-d5 are the same as its unlabeled parent compound. As a retinoid, 9-cis-Retinol acetate is a highly lipophilic prodrug. Following oral administration, it is absorbed from the gastrointestinal tract, incorporated into chylomicrons, and transported via the lymphatic system. It is then converted in the liver to its active metabolite, 9-cis-retinoic acid, primarily by retinol dehydrogenases and retinaldehyde dehydrogenases. The elimination half-life of 9-cis-retinoic acid in humans is relatively short (approx. 2-4 hours). Retinoids are metabolized by CYP26 enzymes (CYP26A1, B1, C1) to more polar metabolites (4-oxo and 4-hydroxy derivatives) and excreted in bile and urine. Retinoids are known teratogens. The labeled 9-cis-Retinol acetate-d5 is used as an internal standard in PK studies to precisely measure these low-abundance compounds via mass spectrometry.
|
| Toxicity/Toxicokinetics |
9-cis-Retinol acetate-d5 is a research-grade chemical and not intended for human use. Its toxicity is inferred from its unlabeled parent compound, which belongs to the class of retinoids. Retinoids, including the active metabolite 9-cis-retinoic acid, are potent teratogens, meaning they can cause severe birth defects if administered to pregnant women. They are also associated with dose-dependent toxicities in humans, including dry skin (mucocutaneous effects), headache, increased liver enzymes (transaminitis), and hypertriglyceridemia. Chronic exposure can lead to skeletal hyperostosis. The acute toxicity of retinol derivatives is relatively low, with an oral LD₅0 in rats >2000 mg/kg. Because of its potential to affect gene expression, standard laboratory safety precautions are critical when handling this compound. Researchers should wear appropriate PPE (gloves, goggles) and avoid exposure to pregnant women. All work should be performed in a well-ventilated area.
|
| References | |
| Additional Infomation |
9-cis-Retinol acetate-d5 is the stable isotope-labeled (deuterated) version of 9-cis-Retinol acetate, a retinal (vitamin A) analog. It is intended for research use only as an internal standard for the accurate quantification of 9-cis-Retinol acetate and its active metabolites (e.g., 9-cis-retinoic acid) in biological samples using LC-MS/MS. The unlabeled compound is a retinoid that acts as a prodrug to activate both retinoic acid receptors (RARs) and retinoid X receptors (RXRs), which are nuclear receptors controlling cell differentiation and proliferation. It is being investigated for the treatment of retinal degenerative diseases such as Leber congenital amaurosis (LCA) and retinitis pigmentosa, as it can restore the visual cycle. It also has potential in cancer prevention and treatment. Due to its teratogenicity, it is strictly a research compound.
|
| Molecular Formula |
C22H32O2
|
|---|---|
| Molecular Weight |
328.488
|
| Exact Mass |
333.271
|
| CAS # |
1217219-62-9
|
| Related CAS # |
9-cis-Retinol acetate;29584-22-3
|
| PubChem CID |
45040693
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
6.3
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
24
|
| Complexity |
596
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
[2H]C1(CCC(C(=C1C([2H])([2H])[2H])/C=C/C(=C/C=C/C(=C/COC(=O)C)/C)/C)(C)C)[2H]
|
| InChi Key |
QGNJRVVDBSJHIZ-XWXKJAMKSA-N
|
| InChi Code |
InChI=1S/C22H32O2/c1-17(9-7-10-18(2)14-16-24-20(4)23)12-13-21-19(3)11-8-15-22(21,5)6/h7,9-10,12-14H,8,11,15-16H2,1-6H3/b10-7+,13-12+,17-9+,18-14+/i3D3,11D2
|
| Chemical Name |
[(2E,4E,6E,8E)-9-[3,3-dideuterio-6,6-dimethyl-2-(trideuteriomethyl)cyclohexen-1-yl]-3,7-dimethylnona-2,4,6,8-tetraenyl] acetate
|
| 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.0442 mL | 15.2212 mL | 30.4423 mL | |
| 5 mM | 0.6088 mL | 3.0442 mL | 6.0885 mL | |
| 10 mM | 0.3044 mL | 1.5221 mL | 3.0442 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
