| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Gilteritinib-d8 targets FLT3 (FMS-like tyrosine kinase 3) and AXL receptors. Gilteritinib is a potent and ATP-competitive inhibitor with IC₅₀ values of 0.29 nM for FLT3 and 0.73 nM for AXL. The compound inhibits FLT3 signaling, which is aberrantly activated in approximately one-third of AML patients through internal tandem duplication (ITD) or tyrosine kinase domain (TKD) mutations. By inhibiting FLT3 and AXL, Gilteritinib blocks downstream signaling pathways including PI3K/AKT, RAS/MEK/ERK, and STAT5, leading to growth arrest and apoptosis in AML cells.
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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 effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1].
In vitro, Gilteritinib demonstrates potent inhibition of FLT3 with an IC₅₀ of 0.29 nM and AXL with an IC₅₀ of 0.73 nM. The compound shows antiproliferative activity against FLT3-mutated AML cell lines including MV4-11 (FLT3-ITD) and MOLM-13 (FLT3-ITD). Gilteritinib induces cell cycle arrest and apoptosis in these cells through inhibition of FLT3 signaling. The deuterated form, Gilteritinib-d8, is used as an internal standard in cell-based studies to accurately quantify Gilteritinib concentrations in cell lysates and culture media. |
| ln Vivo |
Gilteritinib-d8 is used in vivo as a deuterated internal standard for pharmacokinetic studies of Gilteritinib. Gilteritinib has demonstrated antitumor activity in AML xenograft models and is clinically approved (Xospata®) for the treatment of relapsed or refractory AML with FLT3 mutations. In vivo, Gilteritinib inhibits FLT3 phosphorylation and downstream signaling, leading to tumor regression. The deuterated form is employed in LC-MS/MS methods to accurately quantify parent drug concentrations in plasma and tissues during preclinical and clinical pharmacokinetic studies.
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| Enzyme Assay |
In vitro enzyme assays for Gilteritinib involve measuring kinase inhibition using recombinant FLT3 and AXL enzymes. The standard protocol includes incubating the kinase with varying concentrations of Gilteritinib (0.001-1000 nM) and ATP, followed by measurement of phosphorylated substrate using radiometric or fluorescence-based methods. IC₅₀ values are calculated from dose-response curves. For the deuterated form, it is used as an internal standard in LC-MS-based assays to quantify Gilteritinib concentrations. Binding assays may also be performed to assess ATP-competitive binding kinetics.
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| Cell Assay |
Cell-based assays for Gilteritinib are conducted in FLT3-mutated AML cell lines such as MV4-11 (FLT3-ITD) and MOLM-13. Cells are treated with Gilteritinib at concentrations ranging from 0.01-1000 nM for 24-72 hours. Cell viability is assessed using MTT or CellTiter-Glo assays. FLT3 phosphorylation (p-FLT3) and downstream signaling proteins (p-STAT5, p-ERK, p-AKT) are analyzed by western blot. Apoptosis is evaluated by Annexin V staining and caspase activation. The deuterated analog is used as an internal standard for LC-MS/MS quantification of Gilteritinib in cell lysates.
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| Animal Protocol |
In vivo animal experiments for Gilteritinib typically use mouse xenograft models of FLT3-mutated AML. Animals are administered Gilteritinib via oral gavage at doses of 1-30 mg/kg. Tumor volume is measured periodically, and body weight is monitored. Plasma and tumor tissues are collected for pharmacokinetic and pharmacodynamic analysis, including assessment of FLT3 phosphorylation and downstream signaling. The deuterated analog, Gilteritinib-d8, is used as an internal standard in LC-MS/MS methods for the quantification of Gilteritinib in these biological samples.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for Gilteritinib show good oral bioavailability, with a half-life of approximately 10-20 hours in humans. The compound is primarily metabolized by CYP3A4. Gilteritinib-d8 is used as an internal standard for PK studies. The deuterated form provides a mass shift of +8 Da relative to the non-deuterated compound, allowing simultaneous detection by mass spectrometry. Storage: powder at -20°C for 3 years; in solvent at -80°C for 6 months. Purity is typically >98%.
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| Toxicity/Toxicokinetics |
Gilteritinib-d8 is supplied for research use only and is not intended for human administration. The non-deuterated parent compound, Gilteritinib, is a clinically approved drug (Xospata®) for the treatment of relapsed or refractory AML with FLT3 mutations. Common adverse effects include fatigue, fever, anemia, thrombocytopenia, and elevated liver enzymes. The deuterated form is used in trace quantities as an analytical standard. Standard laboratory safety precautions should be followed.
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| References |
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| Additional Infomation |
Gilteritinib-d8 (CAS 2377109-74-3) is a stable isotope-labeled compound used as an internal standard for the quantification of Gilteritinib. It is not intended for therapeutic use. Gilteritinib is a clinically approved FLT3/AXL inhibitor (Xospata®) for the treatment of relapsed or refractory AML with FLT3 mutations. The deuterium labeling is at the piperazine and piperidine ring positions. The compound is for research and analytical applications only.
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| Molecular Formula |
C29H44N8O3
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|---|---|
| Molecular Weight |
552.711465835571
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| Exact Mass |
560.403
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| CAS # |
2377109-74-3
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| Related CAS # |
Gilteritinib;1254053-43-4
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| PubChem CID |
155657789
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| Appearance |
White to yellow solid powder
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| LogP |
3.5
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
40
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| Complexity |
785
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N(C1C=CC(N2CCC(N3C([H])([H])C([H])([H])N(C)C([H])([H])C3([H])[H])CC2)=C(OC)C=1)C1=NC(NC2CCOCC2)=C(CC)N=C1C(=O)N
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| InChi Key |
GYQYAJJFPNQOOW-DBVREXLBSA-N
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| InChi Code |
InChI=1S/C29H44N8O3/c1-4-23-28(31-20-9-17-40-18-10-20)34-29(26(33-23)27(30)38)32-21-5-6-24(25(19-21)39-3)37-11-7-22(8-12-37)36-15-13-35(2)14-16-36/h5-6,19-20,22H,4,7-18H2,1-3H3,(H2,30,38)(H2,31,32,34)/i13D2,14D2,15D2,16D2
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| Chemical Name |
6-ethyl-3-[3-methoxy-4-[4-(2,2,3,3,5,5,6,6-octadeuterio-4-methylpiperazin-1-yl)piperidin-1-yl]anilino]-5-(oxan-4-ylamino)pyrazine-2-carboxamide
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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 | 1.8093 mL | 9.0463 mL | 18.0927 mL | |
| 5 mM | 0.3619 mL | 1.8093 mL | 3.6185 mL | |
| 10 mM | 0.1809 mL | 0.9046 mL | 1.8093 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.
| In Vitro |
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In vitroactivity:In vitro, among the 78 tyrosine kinases tested, Gilteritinib at 1 nM concentration inhibits FLT3, leukocyte tyrosine kinase (LTK), anaplastic lymphoma kinase (ALK), and AXL kinases by over 50% with an IC50value of 0.29 nM for FLT3, and is about 800-fold more potent than for c-KIT.Gilteritinib inhibits eight of the 78 tested kinases by over 50% at concentrations of either 1 nM (FLT3, LTK, ALK, and AXL) or 5 nM (TRKA, ROS, RET, and MER). The IC50values are 0.29 nM for FLT3 and 0.73 nM for AXL. Gilteritinib inhibits FLT3 at an IC50that is approximately 800-fold more potent than the concentration required to inhibit c-KIT (230 nM). The antiproliferative activity of Gilteritinib is evaluated against MV4-11 and MOLM-13 cells, which endogenously express FLT3-ITD. After 5 days of treatment, Gilteritinib inhibits the growth of MV4-11 and MOLM-13 cells with mean IC50s of 0.92 nM (95% CI: 0.23-3.6 nM) and 2.9 nM (95% CI: 1.4-5.8 nM), respectively. Growth suppression of MV4-11 cells is accompanied by inhibition of FLT3 phosphorylation. Relative to vehicle control cells, phosphorylated FLT3 levels are 57%, 8%, and 1% after 2 h of treatment with 0.1 nM, 1 nM, and 10 nM Gilteritinib, respectively. In addition, doses as low as 0.1 nM or 1 nM result in the suppression of phosphorylated ERK, STAT5, and AKT, all of which are downstream targets of FLT3 activation. To investigate the effects of Gilteritinib on AXL inhibition, MV4-11 cells that expressed exogenous AXL are treated with Gilteritinib. At concentrations of 1 nM, 10 nM, and 100 nM for 4 h, Gilteritinib treatment decreases phosphorylated AXL levels by 38%, 29%, and 22%, respectively. Kinase Assay:Gilteritinib was tested in a panel of 78 tested kinases using ATP concentrations that are approximately equal to the Kmvalue for each kinase in a TK-ELISA or off-chip mobility shift assay. Initially, two concentrations of Gilteritinib (1 nM and 5 nM) are tested to assess each compound’s inhibitory effect on TK activity. Further studies are then conducted using a dose range of Gilteritinib to determine IC50values for kinases in which activity is inhibited by >50% with 1 nM Gilteritinib as well as for c-KIT. TK-ELISA and MSA assays are used to conduct IC50studies for FLT3, LTK, AXL, and c-KIT; the HTRF KinEASE-TK assay is performed to assess the IC50value of echinoderm microtubule-associated protein-like 4-ALK (EML4-ALK). Cell Assay:MV4-11 cells treated with DMSO or increasing concentrations of gilteritinib are incubated for 2 hours. Immunoprecipitation and immunoblot for phosphorylated FLT3 and total FLT3 are performed. |
| In Vivo |
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| References |
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