| Size | Price | Stock | Qty |
|---|---|---|---|
| 2mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| Other Sizes |
|
Purity: ≥98%
Allitinib (formerly also known as AST1306; AST-1306) is a novel, potent, selective and covalent / irreversible inhibitor of EGFR and ErbB2 with potential antitumor activity. With IC50s of 0.5 nM and 3 nM, it inhibits both EGFR and ErbB2. It is also effective against the mutation EGFR T790M/L858R. Compared to other kinases, it is more potent against cells that overexpress ErbB2, and it is 3000-fold more selective for the ErbB family. In both cell-free and cell-based systems, AST-1306 suppresses the enzymatic activities of EGFR and ErbB2, as well as EGFR resistant mutants.
| Targets |
EGFR (IC50 = 0.5 nM); ErbB2 (IC50 = 3 nM); EGFRL858R/T790M (IC50 = 12 nM); ErbB4 (IC50 = 0.8 nM)
|
|---|---|
| ln Vitro |
AST1306 (AST-1306; 0.19-6.25 μM; 72 hours) inhibits the growth of HIH3T3-EGFR T790M/L858R cells significantly and in a concentration-dependent manner[1].
AST1306 prevents A549 cells, Calu-3 cells, and SK-OV-3 cells from activating tyrosine kinases and downstream signaling pathways. A549 cells' EGFR phosphorylation is significantly and dose-dependently inhibited by AST1306[1]. AST1306 (0.1, 0.5, 1.0, 5.0 μM) can significantly slow down the growth of both tumor cells on soft agar, with SK-OV-3 cells showing significantly greater sensitivity than A549 cells[1]. AST1306 (0.001-1.0 μM; 4 hours) exhibits a selectivity of over 3000-fold for kinases belonging to the ErbB family as opposed to other kinase families[1]. AST1306 has a 12 nM IC50 value and effectively suppresses the EGFR T790M/L858R double mutant[1]. |
| ln Vivo |
AST1306 (AST-1306; p.o.; 25-100 mg/kg; twice daily; for 28 days) significantly suppresses tumor growth in SK-OV-3 and Calu-3 xenograft models[1].
|
| Enzyme Assay |
Tyrosine kinase activities are measured in 96-well ELISA plates that have been precoated with 20 μg/mL Poly (Glu,Tyr)4:1. Initially, each well receives 80 μL of a diluted 5 μM ATP solution in kinase reaction buffer, which includes 50 mM HEPES pH 7.4, 20 mM MgCl2, 0.1 mM MnCl2, 0.2 mM Na3VO4, and 1 mM DTT. Next, different AST-1306 concentrations are added to each reaction well, with 10 μL of 1% DMSO (v/v) serving as the negative control. The addition of diluted purified tyrosine kinase proteins in 10 μL of kinase reaction buffer solution then starts the kinase reaction. At every concentration, duplicate experiments are carried out. The plate is incubated for 60 minutes at 37 °C. Following this, it is cleaned three times using phosphate buffered saline (PBS) that contains 0.1% Tween 20 (T-PBS). Subsequently, 100 μL of diluted anti-phosphotyrosine antibody (PY99, 1:500 dilution) in T-PBS containing 5 mg/mL BSA is added. The plate is washed three times as before after 30 min of incubation at 37 °C. Goat anti-mouse IgG conjugated with horseradish peroxidase (100 μL) diluted 1:2000 in T-PBS with 5 mg/mL BSA added is added. After a 30-minute reincubation period at 37 °C, the plate is cleaned with PBS. At last, samples are incubated at room temperature until color emerges after 100 μL of a solution containing 0.03% H2O2 and 2 mg/mL o-phenylenediamine in 0.1 M citrate buffer, pH 5.5, is added. After adding 50 μL of 2 M H2SO4, the reaction is stopped, and the plate's wavelength is measured at 490 nm with a multi-well spectrophotometer. The following formula is used to get the inhibition rate (%): [1-(A490 treated /A490 control)] ×100%. The Logit method is used to calculate the IC50 values, which are derived from the outcomes of at least three independent tests.
|
| Cell Assay |
Proliferation of cells (Calu-3, A-549 cell line, et al.) is assessed using the SRB (Sulforhodamine B) assay. To sum up, cells are cultivated for 24 hours after being seeded into 96-well plates. Following treatment, the cells are grown for an additional 72 hours at progressively higher concentrations of AST-1306. Up until the experiment's conclusion, the medium stays unaltered. After using 10% precooled trichloroacetic acid (TCA) for an hour at 4 °C, the cells are stained for 15 minutes at room temperature using 100 μL of a 4 mg/mL SRB solution in 1% acetic acid. The cells are then rapidly washed five times with 1% acetic acid after the SRB is removed. Following the process of air drying the cells, 150 μL of 10 mM Tris base is used to dissolve the protein-bound dye, which is then measured at 515 nm using a multiwell spectrophotometer for five minutes. (1 - A515 treated/A515 control) × 100% is the formula used to determine the inhibition rate on cell proliferation. By using the Logit method, the IC50 value is calculated based on the findings of a minimum of three independent tests.
|
| Animal Protocol |
Nude mice with SK-OV-3 and Calu-3 tumors[1]
25, 50, 100 mg/kg p.o; twice daily; for 28 days |
| References |
| Molecular Formula |
C24H18CLFN4O2
|
|---|---|
| Molecular Weight |
448.88
|
| Exact Mass |
448.11
|
| Elemental Analysis |
C, 64.22; H, 4.04; Cl, 7.90; F, 4.23; N, 12.48; O, 7.13
|
| CAS # |
897383-62-9
|
| Related CAS # |
Allitinib tosylate;1050500-29-2
|
| Appearance |
Solid powder
|
| SMILES |
C=CC(=O)NC1=CC2=C(C=C1)N=CN=C2NC3=CC(=C(C=C3)OCC4=CC(=CC=C4)F)Cl
|
| InChi Key |
MVZGYPSXNDCANY-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C24H18ClFN4O2/c1-2-23(31)29-17-6-8-21-19(11-17)24(28-14-27-21)30-18-7-9-22(20(25)12-18)32-13-15-4-3-5-16(26)10-15/h2-12,14H,1,13H2,(H,29,31)(H,27,28,30)
|
| Chemical Name |
N-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazolin-6-yl]prop-2-enamide
|
| Synonyms |
Allitinib; AST1306; ALS-1306; ALS1306; AST 6; AST-6; AST-1306; Allitinib free base; AST 1306; ALS1306; AST6
|
| 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) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
|
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2278 mL | 11.1388 mL | 22.2777 mL | |
| 5 mM | 0.4456 mL | 2.2278 mL | 4.4555 mL | |
| 10 mM | 0.2228 mL | 1.1139 mL | 2.2278 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
COA
