| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| Other Sizes |
| Targets |
IC50: BTK[1]
Dihydrodiol-Ibrutinib (PCI-45227) selectively targets Bruton's tyrosine kinase (BTK), a non-receptor tyrosine kinase that plays a critical role in B-cell receptor (BCR) signaling, which is essential for the development, survival, and proliferation of B cells. As a metabolite of Ibrutinib, it shares the same target, though its inhibitory activity is significantly lower. The compound binds to the ATP-binding pocket of BTK, inhibiting its kinase activity and downstream signaling pathways. Its formation represents a major metabolic pathway for Ibrutinib inactivation, and its presence in circulation contributes to the overall pharmacological profile of Ibrutinib therapy. |
|---|---|
| ln Vitro |
Dihydrodiol-Ibrutinib demonstrates in vitro inhibitory activity towards BTK, but this activity is approximately 15 times lower than that of its parent compound, Ibrutinib. This reduced potency indicates that while it is pharmacologically active, its direct contribution to BTK inhibition is less significant compared to Ibrutinib. The compound's in vitro activity is characterized by its ability to bind to BTK and inhibit its kinase activity in a dose-dependent manner. Its IC50 for BTK inhibition is significantly higher than that of Ibrutinib, reflecting its status as a less active metabolite.
|
| ln Vivo |
Dihydrodiol-Ibrutinib is the primary circulating metabolite of Ibrutinib and exhibits in vivo activity. It has an extended half-life of 6-11 hours, which is longer than that of the parent drug, and its concentrations in plasma can be up to twice as high as those of Ibrutinib. This extended exposure and higher concentration contribute to the overall pharmacodynamic effect of Ibrutinib therapy, despite its lower intrinsic potency. The metabolite-to-parent (M/P) ratio is an important parameter in pharmacokinetic studies, and the presence of this active metabolite is a key consideration in the clinical pharmacology of Ibrutinib. The in vivo activity of Dihydrodiol-Ibrutinib is assessed by correlating its exposure with the inhibition of BTK signaling in target tissues.
|
| Enzyme Assay |
The in vitro enzyme/receptor binding (non-cell-based) assay for Dihydrodiol-Ibrutinib involves assessing its inhibition of BTK kinase activity in a cell-free system. Recombinant BTK is incubated with the compound, ATP, and a substrate peptide. Kinase activity is measured by detecting the phosphorylation of the substrate using techniques such as radiometric assays, fluorescence-based assays, or ELISA. The IC50 value, which is approximately 15 times higher than that of Ibrutinib, is determined from dose-response curves. Selectivity profiling against other kinases may also be performed to confirm its specificity. The compound's binding affinity to BTK can be assessed using surface plasmon resonance (SPR) or other biophysical methods.
|
| Cell Assay |
The in vitro cell-based assay for Dihydrodiol-Ibrutinib involves treating BTK-dependent cell lines, such as B-cell lines, with the compound and measuring the inhibition of BTK autophosphorylation and downstream signaling. Cells are treated with varying concentrations of the compound, and the phosphorylation of BTK at Tyr223 and its downstream substrates is assessed by Western blotting or ELISA. The compound's cellular potency is determined by its ability to inhibit these signaling events. The inhibition of BCR-induced calcium flux and proliferation in B cells can also be measured to assess the functional consequences of BTK inhibition. The reduced potency compared to Ibrutinib is reflected in the higher concentrations required to achieve similar levels of inhibition.
|
| Animal Protocol |
In vivo animal experiments for Dihydrodiol-Ibrutinib are typically conducted as part of pharmacokinetic and pharmacodynamic studies of Ibrutinib. In these studies, animals are administered Ibrutinib, and the concentrations of both Ibrutinib and its Dihydrodiol metabolite (PCI-45227) are measured in plasma over time using sensitive LC-MS/MS methods. The metabolite-to-parent (M/P) ratio is calculated to understand the extent of metabolism. The compound's in vivo activity is assessed by correlating its exposure with the inhibition of BTK signaling in target tissues, such as spleen or lymph nodes. The pharmacokinetic profile of the metabolite, including its half-life and exposure, is characterized in these studies.
|
| ADME/Pharmacokinetics |
Dihydrodiol-Ibrutinib is the primary pharmacologically active metabolite of Ibrutinib. It is formed via CYP3A4-mediated metabolism and has a longer half-life (6-11 hours) than Ibrutinib. Its plasma concentrations can be up to twice as high as those of the parent drug. The compound has a molecular weight of 474.5 g/mol and a molecular formula of C25H26N6O4. It is soluble in DMSO at 100 mg/mL. In vivo formulation can be prepared using 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline. The compound is typically stored as a powder at -20°C for up to 3 years.
|
| Toxicity/Toxicokinetics |
Dihydrodiol-Ibrutinib is considered a metabolite of Ibrutinib and its toxicological profile is evaluated as part of the overall safety assessment of Ibrutinib. As a research compound, its safety profile is evaluated in standard cytotoxicity and acute toxicity assays. The compound is classified for research use only and is not intended for human therapeutic use. Comprehensive toxicological characterization would be required prior to any clinical development. The compound is typically handled with standard laboratory safety precautions.
|
| References | |
| Additional Infomation |
The metabolites of ibrutinib; their structures are described in the first article.
Dihydrodiol-Ibrutinib (PCI-45227, CAS 1654820-87-7) is a dihydrodiol active metabolite of Ibrutinib. It has a molecular weight of 474.51 and a molecular formula of C25H26N6O4. It is primarily used as a reference standard in pharmacokinetic studies and for research purposes to understand the metabolism and disposition of Ibrutinib. It is not approved for clinical use and is available only for research purposes. The IC50 of the inhibitory effect of PCI-45227 was reported to be 4555 ng/mL, which is 415 times the average steady-state plasma concentration observed in humans dosed with 560 mg/day. The compound's chemical name is 1-[3-[4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]-2,3-dihydroxypropan-1-one. |
| Molecular Formula |
C25H26N6O4
|
|---|---|
| Molecular Weight |
474.51
|
| Exact Mass |
474.201
|
| CAS # |
1654820-87-7
|
| PubChem CID |
91971609
|
| Appearance |
White to off-white solid powder
|
| Density |
1.5±0.1 g/cm3
|
| Boiling Point |
790.3±60.0 °C at 760 mmHg
|
| Flash Point |
431.7±32.9 °C
|
| Vapour Pressure |
0.0±2.9 mmHg at 25°C
|
| Index of Refraction |
1.723
|
| LogP |
1.94
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
35
|
| Complexity |
701
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
C1C[C@H](CN(C1)C(=O)C(CO)O)N2C3=NC=NC(=C3C(=N2)C4=CC=C(C=C4)OC5=CC=CC=C5)N
|
| InChi Key |
NWKPMPRXJGMTKQ-DIAVIDTQSA-N
|
| InChi Code |
InChI=1S/C25H26N6O4/c26-23-21-22(16-8-10-19(11-9-16)35-18-6-2-1-3-7-18)29-31(24(21)28-15-27-23)17-5-4-12-30(13-17)25(34)20(33)14-32/h1-3,6-11,15,17,20,32-33H,4-5,12-14H2,(H2,26,27,28)/t17-,20?/m1/s1
|
| Chemical Name |
1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]-2,3-dihydroxypropan-1-one
|
| 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) |
DMSO: 100 mg/mL (210.74 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.27 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.27 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.27 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1074 mL | 10.5372 mL | 21.0744 mL | |
| 5 mM | 0.4215 mL | 2.1074 mL | 4.2149 mL | |
| 10 mM | 0.2107 mL | 1.0537 mL | 2.1074 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
