TAS-0728 is a potent, selective, covalent, orally bioavailable HER2 inhibitor with IC50 of 36 nM. TAS0728 bound to HER2 kinase, the inhibitory activity was not affected by a high ATP concentration. The inhibitory activity of TAS0728 was unaffected by high ATP concentrations when it bound to HER2 kinase. In xenograft model tumor tissues as well as HER2-amplified breast cancer cells, TAS0728 demonstrated strong and long-lasting inhibition of HER2, HER3, and downstream effector phosphorylation, which culminated in apoptosis. In mouse xenograft models with tumors that were dependent on the HER2 signal, TAS0728 caused tumor regression. In a mouse model of peritoneal dissemination with cancer cells driven by the HER2, it showed a benefit to survival without any obvious toxicity.
Physicochemical Properties
| Molecular Formula | C26H32N8O3 |
| Molecular Weight | 504.584084510803 |
| Exact Mass | 504.26 |
| Elemental Analysis | C, 61.89; H, 6.39; N, 22.21; O, 9.51 |
| CAS # | 2088323-16-2 |
| Related CAS # | 2088323-16-2 |
| PubChem CID | 130275856 |
| Appearance | White to off-white solid powder |
| LogP | 1.7 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 37 |
| Complexity | 869 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | CC1=C(C=CC(=C1C)NC(=O)C2=NN(C3=NC=NC(=C23)N)[C@@H]4CCCN(C4)C(=O)C=C)CC(=O)N(C)C |
| InChi Key | JCCIICHPRAAMGK-GOSISDBHSA-N |
| InChi Code | InChI=1S/C26H32N8O3/c1-6-20(35)33-11-7-8-18(13-33)34-25-22(24(27)28-14-29-25)23(31-34)26(37)30-19-10-9-17(15(2)16(19)3)12-21(36)32(4)5/h6,9-10,14,18H,1,7-8,11-13H2,2-5H3,(H,30,37)(H2,27,28,29)/t18-/m1/s1 |
| Chemical Name | 4-amino-N-[4-[2-(dimethylamino)-2-oxoethyl]-2,3-dimethylphenyl]-1-[(3R)-1-prop-2-enoylpiperidin-3-yl]pyrazolo[3,4-d]pyrimidine-3-carboxamide |
| Synonyms | TAS 0728; TAS0728; TAS-0728 |
| 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) |
Biological Activity
| Targets | HER4 (IC50 = 8.5 nM); HER2 (IC50 = 13 nM); EGFR (IC50 = 65 nM); BMX (IC50 = 4.9 nM); BLK (IC50 = 31 nM); JAK3 (IC50 = 33 nM); SLK (IC50 = 25 nM); LOK (IC50 = 86 nM) |
| ln Vitro | TAS0728 selectively inhibits the kinase activity of HER2 by covalently binding to it at C805. High ATP concentrations have no effect on TAS0728's inhibitory activity once it binds to HER2 kinase. TAS0728 exhibits superior HER2 specificity compared to wild-type EGFR. Both wild-type and mutant HER2 have their phosphorylation significantly inhibited by TAS0728. The phosphorylation of HER2, HER3, and downstream effectors is robustly and persistently inhibited by TAS0728, which causes HER2-amplified breast cancer cells to undergo apoptosis. [1] |
| ln Vivo | TAS0728 causes apoptosis in tumor tissues of a xenograft model by demonstrating strong and long-lasting inhibition of the phosphorylation of HER2, HER3, and downstream effectors. In mouse models of xenografts with tumors driven by the HER2 signal, TAS0728 causes tumor regression; in a mouse model of peritoneal dissemination with cancer cells driven by the HER2 signal, it shows a benefit to survival without obvious toxicity.[1] |
| Enzyme Assay | Kinase profiling is a technique used by Carna Biosciences to quantify the inhibitory activity of HER2 kinase. Based on HER2's in vitro peptide substrate phosphorylation activity, the 50% inhibitory concentration (IC50 value) of TAS0728 is calculated. Three separate studies are carried out. Reaction Biology Corporation utilizes the Kinase Panel Assay to test TAS0728's inhibitory activity against 386 or 374 kinases. The assays are carried out in duplicate mode with TAS0728 concentrations of 0.1, 1, and 10 μmol/L. 10 μmol/L ATP is present when reactions are occurring. |
| Cell Assay | In 6-well plates, cells are sown, and they are cultured for an entire night. For three hours, compounds are added to the culture medium at the indicated concentrations. After that, the medium is harvested for Western blot analysis. The MCF10A cell line is used for studies in which 12-well plates are seeded with cells and left overnight to culture. After adding TAS0728 to the culture medium, it is incubated for three hours. To carry out a Western blot analysis, the cells are extracted. 100-mm dishes are used to seed and culture SK-BR-3 cells for the duration of the pharmacodynamics time-course study. Add TAS0728, then let it sit for three or forty-eight hours. To carry out a Western blot analysis, the cells are extracted. |
| Animal Protocol |
6-week-old male nude mice (BALB/cAJcl-nu/nu) 7.5 mg/kg, 15 mg/kg, 30 mg/kg, 60 mg/kg Oral gavage |
| References |
[1]. TAS0728, A Covalent-binding, HER2-selective Kinase Inhibitor Shows Potent Antitumor Activity in Preclinical Models. Molecular cancer therapeutics. 2019;18(4):733-42. Epub 2019/02/23. |
Solubility Data
| Solubility (In Vitro) |
DMSO: 100~125 mg/mL (198.2~247.7 mM） Ethanol: ~5 mg/mL (~9.9 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.12 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 (4.12 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (4.12 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9818 mL | 9.9092 mL | 19.8185 mL | |
| 5 mM | 0.3964 mL | 1.9818 mL | 3.9637 mL | |
| 10 mM | 0.1982 mL | 0.9909 mL | 1.9818 mL |