| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| Other Sizes |
TL02-59 is a novel, potent and orally bioavailable Src-family kinase Fgr inhibitor with an IC50 of 0.03 nM. TL02-59 also inhibits Lyn and Hck with IC50s of 0.1 nM and 160 nM, respectively. TL02-59 potently suppresses acute myelogenous leukemia (AML) cell growth.
| Targets |
TL02-59 is a selective inhibitor of the myeloid Src-family kinase Fgr (IC50 = 0.031 nM).
It also potently inhibits Lyn (IC50 = 0.10 nM), and with lower potency inhibits Hck (IC50 = 160 nM), Flt3-ITD (IC50 = 440 nM), Fes (IC50 = 290 nM), Syk (IC50 = 470 nM), p38α (IC50 = 126 nM), and Taok3 (IC50 = 509 nM) [1]. |
|---|---|
| ln Vitro |
TL02-59 (0.1-1000 nM; 6 hours) substantially reduces Fgr autophosphorylation in TF-1 cells, with moderate inhibition at 0.1-1 nM and total inhibition beyond 10 nM. Hck, Lyn and Flt3 are effective in regulating nM in the proliferation of TL02-59 primary AML cell lines and induce cell inhibition [1]. TL02-59 is suppressed in primary AML cell samples in the range of 100 to 1000 nM [1].
TL02-59 potently inhibited the proliferation of the FLT3-ITD+ AML cell lines MV4-11 (IC50 = 0.78 nM) and MOLM-14 (IC50 = 6.6 nM), but had minimal effect on THP-1 cells (no significant inhibition at 1 μM). [1] TL02-59 induced apoptosis in MV4-11 and MOLM-14 cells at 100 nM, an effect more potent than the Flt3 inhibitor tandutinib at the same concentration, while it did not induce apoptosis in THP-1 cells. [1] Treatment of primary bone marrow samples from 26 AML patients with TL02-59 resulted in a wide range of growth inhibitory responses (IC50 values ranging from 77 nM to >3000 nM). Sensitivity strongly correlated with high expression levels of the myeloid Src-family kinases (Fgr, Hck, Lyn) and Syk, but was independent of FLT3 mutation status. The four most sensitive samples were FLT3 wild-type. [1] In TF-1 myeloid cells engineered to express individual kinases, TL02-59 treatment (0.1-1000 nM) potently and selectively inhibited Fgr autophosphorylation (complete inhibition at ≥10 nM), while inhibition of Hck, Lyn, and Flt3-ITD required higher concentrations (100-1000 nM). [1] Lentiviral shRNA-mediated knockdown of Fgr expression in MV4-11 cells suppressed cell proliferation, corroborating Fgr as a relevant target. [1] |
| ln Vivo |
AML cells in the spleen and peripheral blood were totally eradicated by TL02-59 (package insert; 1 mg/kg and 10 mg/kg; for three weeks) while cell involvement in the AML model was markedly inhibited [1]. For intravenous and intravenous hour formulations, the t1/2 of 59 was 5.7 hours and 6.5 hours, respectively [1].
In a mouse xenograft model of AML using MV4-11 cells, oral administration of TL02-59 at 10 mg/kg once daily for 21 days completely eradicated leukemic cells (human CD45+/CD33+) from the spleen and peripheral blood, and reduced bone marrow engraftment by 60%. At 1 mg/kg, it reduced spleen engraftment by 50%, peripheral blood involvement by 70%, and bone marrow engraftment by 20%. In contrast, the Flt3 inhibitor sorafenib (10 mg/kg) had no significant effect on bone marrow or spleen engraftment, although it reduced peripheral blood counts. [1] Immunohistochemistry of spleen and bone marrow sections confirmed the near-complete elimination of MV4-11 cells (stained for human CD45) in mice treated with TL02-59 at 10 mg/kg. [1] |
| Enzyme Assay |
In vitro kinase inhibition assays were performed using a FRET-based method. Recombinant kinase domains or full-length kinases were incubated with TL02-59 across a range of concentrations. The ATP concentration in each reaction was set to the Km value for the respective kinase. Dose-response curves were generated, and IC50 values were calculated by nonlinear regression analysis. [1]
The KINOMEscan profiling assay, a competitive binding assay, was used to assess the kinase selectivity profile of TL02-59 at a screening concentration of 1 μM against 456 human kinases. The percentage of control binding (residual probe bound) was measured for each kinase. [1] |
| Cell Assay |
Western Blot Analysis [1]
Cell Types: TF-1 bone marrow cells Tested Concentrations: 0.1, 1, 10, 100, 1000 nM Incubation Duration: 6 hrs (hours) Experimental Results: Inhibition of Fgr autophosphorylation in TF-1 cells. For cell proliferation assays, AML cell lines or primary bone marrow cells were seeded and treated with a range of concentrations of TL02-59 or DMSO control. After 72 hours of incubation, cell viability was measured using a resazurin-based metabolic assay. Dose-response curves were plotted, and IC50 values were determined. [1] For apoptosis assays, cells were treated with inhibitors or DMSO for 72 hours. Caspase activity was measured using a luminescent substrate and normalized to cell viability from a parallel assay to determine the ratio of apoptosis to viability. [1] For kinase activity assessment in cells, TF-1 cells expressing specific kinases were treated with TL02-59 for 6 hours. Cells were lysed, and the kinase of interest was immunoprecipitated from the clarified lysate. Immunoprecipitates were resolved by SDS-PAGE and immunoblotted with antibodies against phosphotyrosine (for Flt3) or the phosphorylated activation loop tyrosine (for Src-family kinases). Total kinase levels were verified by blotting with kinase-specific antibodies. [1] For gene expression analysis in primary AML samples, total RNA was isolated, reverse transcribed, and analyzed by quantitative real-time PCR (qPCR) using gene-specific primers. Expression levels were calculated relative to GAPDH and then normalized to the mean expression of all target kinases within each sample. [1] |
| Animal Protocol |
Animal/Disease Models: NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice and human MV4-11 AML cells [1]: 1 and 10 mg/kg
Route of Administration: oral; continued for three weeks Experimental Results: Elimination of AML in mouse model AML cells in the spleen and peripheral blood, while Dramatically suppressing bone marrow involvement. NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) immunocompromised mice were injected via the tail vein with 10^7 MV4-11 human AML cells suspended in PBS. After 14 days to allow engraftment, mice were randomly divided into treatment groups (n=8). Treatments were administered once daily for 21 days by oral gavage. The treatment groups received: vehicle control, TL02-59 at 1 mg/kg, TL02-59 at 10 mg/kg, or sorafenib at 10 mg/kg. The formulation details for the inhibitors were not specified. At the end of the treatment period, mice were euthanized. Bone marrow, spleen, and peripheral blood were collected. The presence of human AML cells was quantified by flow cytometry using antibodies against human CD45 and CD33. For histology, spleen and bone marrow sections were fixed and stained with an antibody against human CD45. [1] |
| ADME/Pharmacokinetics |
Pharmacokinetic studies in mice showed that TL02-59 has oral bioavailability and a plasma half-life of approximately 6 hours. [1]
|
| References | |
| Additional Infomation |
TL02-59 is an N-phenylbenzamide kinase inhibitor. [1] Its anti-AML activity is mainly achieved by potently inhibiting the Src family kinase Fgr, which has not been previously considered in AML treatment. [1] Its efficacy has been observed in AML models independent of FLT3 mutation status, suggesting that it may have potential application value in AML patient subgroups that overexpress Fgr and other myeloid Src family kinases. [1] Analysis of TCGA data suggests that Fgr overexpression may occur in about one-third or more of AML cases. [1]
|
| Molecular Formula |
C₃₂H₃₄F₃N₅O₄
|
|---|---|
| Molecular Weight |
609.638678073883
|
| Exact Mass |
609.256
|
| CAS # |
1315330-17-6
|
| Related CAS # |
TL02-59 dihydrochloride;2415263-06-6
|
| PubChem CID |
71254350
|
| Appearance |
White to off-white solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
630.5±55.0 °C at 760 mmHg
|
| Flash Point |
335.1±31.5 °C
|
| Vapour Pressure |
0.0±1.8 mmHg at 25°C
|
| Index of Refraction |
1.603
|
| LogP |
5.22
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
44
|
| Complexity |
923
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
AMHWQBGAKJESFB-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C32H34F3N5O4/c1-5-39-10-12-40(13-11-39)18-22-8-9-23(15-25(22)32(33,34)35)38-30(41)21-7-6-20(2)27(14-21)44-31-24-16-28(42-3)29(43-4)17-26(24)36-19-37-31/h6-9,14-17,19H,5,10-13,18H2,1-4H3,(H,38,41)
|
| Chemical Name |
3-[(6,7-Dimethoxy-4-quinazolinyl)oxy]-N-[4-[(4-ethyl-1-piperazinyl)methyl]-3-(trifluoromethyl)phenyl]-4-methylbenzamide
|
| Synonyms |
TL02-59 TL02 59 TL0259
|
| 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 : ~125 mg/mL (~205.04 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.41 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 20.8 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: ≥ 2.08 mg/mL (3.41 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 20.8 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: ≥ 2.08 mg/mL (3.41 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 | 1.6403 mL | 8.2016 mL | 16.4031 mL | |
| 5 mM | 0.3281 mL | 1.6403 mL | 3.2806 mL | |
| 10 mM | 0.1640 mL | 0.8202 mL | 1.6403 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
