TWS119, a pyrrolopyrimidine compound, is a novel, potent and selective/specific GSK-3β (Glycogen synthase kinase-3β) inhibitor with potential usefulness for in vivo stem cell biology and therapy. It is capable of inducing neuronal differentiation and might be helpful for stem cell biology. It inhibits GSK-3β with an IC50 of 30 nM in a cell-free assay. When using mouse P19 EC cells, it is screened out of a library of pyrrolopyrimidines as a substance that selectively induces neuronal differentiation. TWS119 has a Kd value of 126 nM and strongly binds to GSK-3β. Combining TWS119 and GSK-3β modifies the complex's activity and causes subsequent transcriptional events that result in the induction of neurons. Additionally, TWS119 promotes neuronal differentiation of mESCs via a different mechanism as opposed to the conventional Wnt signaling pathway.
Physicochemical Properties
| Molecular Formula | C18H14N4O2 |
| Molecular Weight | 318.3294 |
| Exact Mass | 318.111 |
| Elemental Analysis | C, 67.92; H, 4.43; N, 17.60; O, 10.05 |
| CAS # | 601514-19-6 |
| Related CAS # | TWS119 TFA;1507095-58-0 |
| PubChem CID | 9549289 |
| Appearance | Off-white to yellow solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 646.0±55.0 °C at 760 mmHg |
| Flash Point | 344.5±31.5 °C |
| Vapour Pressure | 0.0±2.0 mmHg at 25°C |
| Index of Refraction | 1.753 |
| LogP | 3.54 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 24 |
| Complexity | 424 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O(C1=C([H])C([H])=C([H])C(=C1[H])O[H])C1C2C([H])=C(C3C([H])=C([H])C([H])=C(C=3[H])N([H])[H])N([H])C=2N=C([H])N=1 |
| InChi Key | VPVLEBIVXZSOMQ-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C18H14N4O2/c19-12-4-1-3-11(7-12)16-9-15-17(22-16)20-10-21-18(15)24-14-6-2-5-13(23)8-14/h1-10,23H,19H2,(H,20,21,22) |
| Chemical Name | 3-((6-(3-aminophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)phenol |
| Synonyms | TWS-119; TWS 119; TWS119 |
| 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 | GSK-3β (IC50 = 30 nM) |
| ln Vitro | Treatment of a monolayer of P19 cells with 1 μM TWS119 causes 30–40% cells to differentiate specifically into neuronal lineages based on counting of TuJ1 positive cells with correct neuronal morphology (up to 60% neuronal differentiation occurred through the standard EB formation protocol with concomitant TWS119 treatment). Surface plasmon resonance (SPR) measurements of TWS119's strong GSK-3-binding affinity (K D = 126 nM) and IC50 of 30 nM further support this conclusion. [1] WS119 is found to potently induces neuronal differentiation in both mouse embryonal carcinoma and ES cells. [2] TWS119 treatment towards hepatic stellate cells (HSC) leads to reduced b-catenin phosphorylation, induces nuclear translocation of b-catenin, elevates glutamine synthetase production, impedes synthesis of smooth muscle actin and Wnt5a, but promotes the expression of glial fibrillary acidic protein, Wnt10b, and paired-like homeodomain transcription factor 2c.[3] TWS119 causes a sharp up-regulation of the expression of Tcf7, Lef1, and other Wnt target genes like Jun, Ezd7 (encoding Frizzled-7), and Nlk (encoding Nemo-like kinase) in addition to triggering a rapid accumulation of -catenin (mean 6.8-fold increase by densitometry). TWS119 causes a dose-dependent reduction in T cell-specific killing and IFN-g release along with the maintenance of IL-2 production.[4] In polyclonally activated human T cells, treatment with TWS119 induces Wnt signaling, according to a recent study. Unlike control-activated T cells, which develop a CD45RO(+)CD62L(-) effector phenotype in a TWS119 dose-dependent manner, these T cells maintain a native CD45RA(+)CD62L(+) phenotype. Because cell division is prevented by TWS119-induced Wnt signaling, T cell expansion is reduced. Additionally, degranulation and IFN- production in response to T cell activation—which are both indicators of T cell effector function—are impaired. The inability of TWS119-treated T cells to use autocrine IL-2 for expansion may be the cause of the block in T cell division. This is because TWS119 treatment reduces the expression of the IL-2R. [5] |
| ln Vivo | A cell population that expressed low levels of CD44 and high levels of CD62L on the cell surface when 30 mg/kg of TWS119 is administered.[4] |
| Cell Assay | All rats are divided into four groups at random as follows: Sham group rats undergo the same surgical procedure, but the filament is not inserted, and they are given 1 mL of dimethyl sulfoxide (1% DMSO in saline); After MCAO, the rats in the vehicle group receive 1 mL of DMSO. At 4 hours after MCAO, the rats in the rtPA group receive rtPA (10 mg/kg, Actilyse®). At 4 hours after MCAO, the rats in the rtPA+TWS119 group receive intraperitoneal TWS119 (30 mg/kg, dissolved in 1 mL 1% DMSO). |
| Animal Protocol | All rats are divided into four groups at random as follows: Sham group rats undergo the same surgical procedure, but the filament is not inserted, and they are given 1 mL of dimethyl sulfoxide (1% DMSO in saline); After MCAO, the rats in the vehicle group receive 1 mL of DMSO. At 4 hours after MCAO, the rats in the rtPA group receive rtPA (10 mg/kg, Actilyse®). At 4 hours after MCAO, the rats in the rtPA+TWS119 group receive intraperitoneal TWS119 (30 mg/kg, dissolved in 1 mL 1% DMSO). |
| References |
[1]. Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7632-7. [2]. Nat Biotechnol. 2004 Jul;22(7):833-40. |
| Additional Infomation | 3-[[6-(3-aminophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]oxy]phenol is a member of pyrroles. |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~64 mg/mL (201.0 mM) Water: <1 mg/mL Ethanol: <1 mg/mL |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.85 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 (7.85 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. Solubility in Formulation 3: 1% DMSO+30% polyethylene glycol+1% Tween 80: 30mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1414 mL | 15.7070 mL | 31.4139 mL | |
| 5 mM | 0.6283 mL | 3.1414 mL | 6.2828 mL | |
| 10 mM | 0.3141 mL | 1.5707 mL | 3.1414 mL |