| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
Bruton‘s Tyrosine Kinase (BTK). Ibrutinib D5 acts as an irreversible (covalent) and highly selective inhibitor of BTK. It binds covalently to a cysteine residue (Cys481) in the ATP-binding pocket of BTK, blocking its kinase activity. This inhibition prevents the phosphorylation of downstream targets (such as PLCgamma2 and ERK), disrupting B-cell receptor (BCR) signaling, inhibiting B-cell proliferation, migration, and adhesion, and ultimately promoting apoptosis of malignant B-cells.
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| ln Vitro |
Ibrutinib inhibits BTK with an IC50 of 0.5 nM in enzyme activity assays. It is highly selective for BTK over other Tec-family kinases (e.g., BMX, ITK, TEC) and most off-target kinases, though it also inhibits EGFR, JAK3, and BLK at higher concentrations (IC50s 10-100 nM). It covalently binds to BTK, resulting in sustained target inhibition that outlasts the compound's plasma exposure.
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| ln Vivo |
In mouse models of human B-cell malignancies (e.g., CLL, MCL, DLBCL xenografts), oral Ibrutinib (6.25-25 mg/kg once daily) significantly inhibits tumor growth (TGI >80%), induces tumor regression, and improves survival. It inhibits phosphorylation of BTK and PLCgamma2 in tumor tissues, reduces serum levels of pro-inflammatory cytokines (e.g., IL-6, TNFalpha), and decreases B-cell chemotaxis to CXCL12 and CXCL13.
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| Enzyme Assay |
Cell-free BTK kinase assays are performed using recombinant human BTK enzyme (1-5 nM) and a biotinylated substrate peptide. The reaction mix contains 50 mM HEPES (pH 7.5), 10 mM MgCl2, 2 mM MnCl2, 1 mM DTT, 0.01% BSA, and 10 uM ATP (or 100 uM ATP). Varying concentrations of Ibrutinib D5 (0.01-1000 nM) are added. After 60 min at 30degC, the reaction is quenched, and substrate phosphorylation is measured using an anti-phosphotyrosine antibody and time-resolved fluorescence (TR-FRET). IC50 values are calculated.
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| Cell Assay |
TMD-8 human diffuse large B-cell lymphoma (DLBCL) cells (BCR-dependent) are cultured in RPMI-1640 with 10% FBS. Cells (2-5 × 10⁵ cells/mL) are treated with Ibrutinib D5 (0.1-1000 nM) or vehicle for 2-24 h at 37degC, 5% CO2. Cells are lysed, and BTK autophosphorylation at Tyr223 (pBTK) and phosphorylation of downstream targets (pPLCgamma2, pERK, pAKT) are measured by Western blotting. The cellular IC50 for pBTK inhibition is calculated. Cell viability is measured by CellTiter-Glo after 48-72 h of treatment.
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| Animal Protocol |
Female SCID-beige mice (6-8 weeks) bearing subcutaneous TMD-8 xenografts (tumor volume 150-200 mm3) are randomized into treatment groups (n=8-10). Ibrutinib (6.25, 12.5, 25 mg/kg) or vehicle is administered orally once daily for 14-28 days. Tumor volumes are measured twice weekly. At study termination, plasma and tumors are collected. Tumor BTK and pBTK levels are quantified by Western blot. Plasma Ibrutinib concentrations are measured by LC-MS/MS using Ibrutinib D5 as the internal standard.
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| ADME/Pharmacokinetics |
Ibrutinib D5 is used as an internal standard (IS) for LC-MS/MS bioanalysis. Ibrutinib is rapidly absorbed (Tmax ~1-2 h) with a terminal half-life of 4-8 h. It has high oral bioavailability (~90% in fed state) but is metabolized extensively by CYP3A4 to several active metabolites (e.g., dihydrodiol-ibrutinib, which has ~15-fold lower BTK inhibitory activity). The parent drug is >97% protein bound.
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| Toxicity/Toxicokinetics |
The D5-labeled compound is not for human use. The parent drug Ibrutinib has a boxed warning for severe bleeding (hemorrhage), infections (fatal opportunistic infections), and atrial fibrillation/flutter. Common AEs (≥30%): diarrhea, fatigue, musculoskeletal pain, nausea, rash, bruising (contusion), and thrombocytopenia. Serious AEs: major hemorrhage (4-6%), atrial fibrillation (3-11%), and hypertension.
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| References | |
| Additional Infomation |
Ibrutinib (Imbruvica) was first FDA-approved in 2013 for mantle cell lymphoma (MCL) and has since received approval for CLL/SLL, Waldenström‘s macroglobulinemia, marginal zone lymphoma (MZL), and chronic graft-versus-host disease (cGVHD). Ibrutinib D5 is a research internal standard used in LC-MS/MS bioanalysis for quantifying Ibrutinib in PK/PD studies, therapeutic drug monitoring (TDM), drug-drug interaction studies (especially CYP3A4 inhibitors/inducers), and metabolite profiling.
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| Molecular Formula |
C25H19D5N6O2
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|---|---|
| Molecular Weight |
445.528
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| Exact Mass |
445.227
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| CAS # |
1553977-17-5
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| Related CAS # |
Ibrutinib;936563-96-1;Ibrutinib Racemate;936563-87-0
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| PubChem CID |
73053689
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| Appearance |
White to off-white solid powder
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| LogP |
4.736
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
33
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| Complexity |
678
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| Defined Atom Stereocenter Count |
1
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| SMILES |
[2H]C1=C(C(=C(C(=C1[2H])[2H])OC2=CC=C(C=C2)C3=NN(C4=NC=NC(=C34)N)[C@@H]5CCCN(C5)C(=O)C=C)[2H])[2H]
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| InChi Key |
XYFPWWZEPKGCCK-OZUAZJOXSA-N
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| InChi Code |
InChI=1S/C25H24N6O2/c1-2-21(32)30-14-6-7-18(15-30)31-25-22(24(26)27-16-28-25)23(29-31)17-10-12-20(13-11-17)33-19-8-4-3-5-9-19/h2-5,8-13,16,18H,1,6-7,14-15H2,(H2,26,27,28)/t18-/m1/s1/i3D,4D,5D,8D,9D
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| Chemical Name |
1-[(3R)-3-[4-amino-3-[4-(2,3,4,5,6-pentadeuteriophenoxy)phenyl]pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]prop-2-en-1-one
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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 | 2.2445 mL | 11.2226 mL | 22.4452 mL | |
| 5 mM | 0.4489 mL | 2.2445 mL | 4.4890 mL | |
| 10 mM | 0.2245 mL | 1.1223 mL | 2.2445 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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