| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
The unlabeled parent, cyclosporin A, binds to the intracellular receptor cyclophilin. The cyclosporin-cyclophilin complex then binds to and inhibits the phosphatase activity of protein phosphatase 2B (PP2B), also known as calcineurin, with an IC50 of approximately 5-7 nM. Inhibition of calcineurin prevents the dephosphorylation of nuclear factor of activated T-cells (NFAT), thereby blocking the transcription of IL-2 and other cytokines involved in T-cell activation and proliferation. Cyclosporin A acetate-d4 retains the same target binding affinity due to minimal isotope effect.
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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 impact on a drug's pharmacokinetics and metabolic profile, it has drawn attention [1].
The unlabeled cyclosporin A potently inhibits T-cell activation in mixed lymphocyte reactions (MLR) and mitogen-induced proliferation assays with IC50 values in the low nanomolar to sub-nanomolar range (approx. 0.5-10 nM). It suppresses the production of IL-2, IFN-gamma, and other pro-inflammatory cytokines. Cyclosporin A also inhibits the phosphatase activity of purified calcineurin in vitro with IC50 ~5 nM. The d4 label does not significantly alter these properties. |
| ln Vivo |
Cyclosporin A (CsA) is a potent and efficacious immunosuppressive agent in various animal models of transplantation and autoimmunity. In rodent models, CsA administered at doses of 10-50 mg/kg (oral or i.p.) significantly prolongs the survival of skin, heart, kidney, and liver allografts by inhibiting T-cell-mediated rejection. It also suppresses the development of autoimmune diseases such as collagen-induced arthritis and experimental autoimmune encephalomyelitis (EAE). The d4 internal standard is not used for in vivo pharmacological assessment.
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| Enzyme Assay |
As an internal standard, cyclosporin A acetate-d4 is used in LC-MS/MS method development. The compound is supplied as a solid or in solution (e.g., 100 microg/mL in acetonitrile). Its identity is confirmed by the shift in molecular weight due to the introduction of four deuterium atoms, which provides a distinct m/z ratio from the unlabeled analyte. The recommended storage condition is -20degC, protected from light. Standard solutions for spiking are prepared in methanol or acetonitrile.
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| Cell Assay |
Cyclosporin A acetate-d4 is not directly used in cell-based assays as a functional pharmacological agent; however, the unlabeled parent compound serves as a reference. The standard in vitro protocol for assessing its immunosuppressive activity is the mixed lymphocyte reaction (MLR). Human peripheral blood mononuclear cells (PBMCs) from two unrelated donors are isolated and co-cultured (1 × 10⁵ cells/well each) in 96-well plates. Cyclosporin A is added at concentrations ranging from 0.1-100 nM. After 5-7 days, proliferation is measured by [3H]-thymidine incorporation (0.5 microCi/well added for the final 16-18 hours) before harvesting. Counts per minute (CPM) are measured by liquid scintillation. IC50 values are calculated from the inhibition curve.
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| Animal Protocol |
The unlabeled cyclosporin A is used in animal transplantation studies. A typical protocol: In a rat heterotopic heart transplantation model, donor hearts from Lewis rats are transplanted into the abdominal cavity of recipient rats. Cyclosporin A (10-25 mg/kg/day) is administered subcutaneously or orally for 7-14 days post-transplantation. Graft survival is monitored daily by abdominal palpation to detect cessation of beating. Rejection is confirmed by histopathological examination (endomyocardial biopsy) of the graft. Blood samples are collected for drug level monitoring of CsA using LC-MS/MS with the d4-labeled internal standard. Survival data are presented as Kaplan-Meier curves and compared by log-rank test.
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| ADME/Pharmacokinetics |
The unlabeled cyclosporin A exhibits unpredictable oral bioavailability (approx. 20-30% in humans) due to P-glycoprotein (ABCB1) efflux and intestinal/hepatic metabolism by CYP3A4. The peak blood concentration (Cmax) is reached 1.5-3 hours post-dose. The elimination half-life is 8-24 hours (highly variable), primarily dependent on liver function, as CsA is extensively metabolized in the liver (>99% metabolism). Clearance is predominantly biliary/fecal. Cyclosporin A acetate-d4, as an internal standard, shows identical chromatographic and ionization behavior but is not administered alone for PK profiling.
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| Toxicity/Toxicokinetics |
Cyclosporin A acetate-d4 is a stable isotopologue and poses no significant toxicological risk at the trace concentrations used for analytical purposes (pg/mL-ng/mL). The unlabeled parent drug cyclosporin A, however, is associated with significant dose-limiting toxicities including nephrotoxicity (acute and chronic), hypertension, neurotoxicity (tremor, seizures), hepatotoxicity (cholestasis), hyperlipidemia, gingival hyperplasia, and increased risk of infections and lymphoproliferative disorders. Monitoring blood levels is clinically required to prevent toxicity.
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| References |
[1]. Russak EM, et al. Impact of Deuterium Substitution on the Pharmacokinetics of Pharmaceuticals. Ann Pharmacother. 2019 Feb;53(2):211-246.
[2]. Dalmarco EM, et al. Cyclosporin A inhibits CD11a/CD18 adhesion molecules due to inhibition of TNFalpha and IL-1 beta levels in the mouse model of pleurisy induced by carrageenan. Cell Adh Migr. 2008 Oct-Dec;2(4):231-5. [3]. Borel JF, et al. Effects of the new anti-lymphocytic peptide cyclosporin A in animals. Immunology. 1977 Jun;32(6):1017-25. [4]. Williams, R, et al. Randomised trial comparing FK506 and cyclosporin in prevention of liver allograft rejection. European FK506 Multicentre Liver Study Group. Lancet, 1994, 344(8920), 423-428. [5]. Nicolli A, et al. Interactions of cyclophilin with the mitochondrial inner membrane and regulation of the permeability transition pore, and cyclosporin A-sensitive channel. J Biol Chem. 1996 Jan 26;271(4):2185-92. [6]. Fruman DA, et al. Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3686-90. [7]. Flanagan WM, et al. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A. Nature. 1991 Aug 29;352(6338):803-7. [8]. Handschumacher RE, et al. Cyclophilin: a specific cytosolic binding protein for cyclosporin A. Science. 1984 Nov 2;226(4674):544-7. [9]. Liu J, et al. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell. 1991 Aug 23;66(4):807-15. |
| Additional Infomation |
The d4-labeled cyclosporin A acetate is a deuterium isotopologue of the immunosuppressant cyclosporin A (CsA). CsA revolutionized solid organ transplantation following its FDA approval in 1983. The acetate salt form (typically used for in vivo administration) is stable and soluble in organic solvents. Cyclosporin A is also used off-label for severe rheumatoid arthritis, psoriasis, and other immune-mediated conditions. The deuterium labeling (d4) provides superior compensation for matrix effects in LC-MS/MS compared to structural analogs. Cyclosporin A acetate-d4 is for research use only and not intended for human consumption.
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| Molecular Formula |
C64H109D4N11O13
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| Molecular Weight |
1248.67
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| Related CAS # |
Cyclosporin A;59865-13-3
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| Appearance |
Typically exists as solid 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 | 0.8009 mL | 4.0043 mL | 8.0085 mL | |
| 5 mM | 0.1602 mL | 0.8009 mL | 1.6017 mL | |
| 10 mM | 0.0801 mL | 0.4004 mL | 0.8009 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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