BRD0705 is a novel, potent, paralog selective and orally bioactive glycogen synthase kinase 3α (GSK3α) inhibitor with an IC50 of 66 nM and a Kd of 4.8 μM. GSK3 is a key regulatory kinase in the wingless-type MMTV integration site family (WNT) pathway, is a therapeutic target for many diseases. Potential neoplastic worries are lessened by the GSK3-selective substance BRD0705, which inhibits kinase activity without stabilizing -catenin. In AML cells, BRD0705 induces myeloid differentiation and hinders colony formation, but it has no apparent impact on healthy hematopoietic cells. BRD0705 also prolongs survival in AML mouse models and hinders leukemia initiation. These studies show that paralog-selective GSK3α inhibition is feasible and offers a promising therapeutic method for AML.
Physicochemical Properties
| Molecular Formula | C16H12CLNO4S |
| Molecular Weight | 349.788782119751 |
| Exact Mass | 321.184 |
| Elemental Analysis | C, 74.74; H, 7.21; N, 13.07; O, 4.98 |
| CAS # | 2056261-41-5 |
| Related CAS # | BRD5648;2056261-42-6;(Rac)-BRD0705;1597440-03-3 |
| PubChem CID | 136980453 |
| Appearance | White to light yellow solid powder |
| LogP | 4.2 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 3 |
| Rotatable Bond Count | 2 |
| Heavy Atom Count | 24 |
| Complexity | 561 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | CC[C@@]1(C2=C(NC3=C1C(=O)CC(C3)(C)C)NN=C2)C4=CC=CC=C4 |
| InChi Key | NCKLQXXBRWCYMA-FQEVSTJZSA-N |
| InChi Code | InChI=1S/C20H23N3O/c1-4-20(13-8-6-5-7-9-13)14-12-21-23-18(14)22-15-10-19(2,3)11-16(24)17(15)20/h5-9,12H,4,10-11H2,1-3H3,(H2,21,22,23)/t20-/m0/s1 |
| Chemical Name | (4S)-4-ethyl-7,7-dimethyl-4-phenyl-1,6,8,9-tetrahydropyrazolo[3,4-b]quinolin-5-one |
| Synonyms | BRD0705; BRD-0705; BRD 0705 |
| 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 | GSK3α (IC50 = 66 nM); GSK3α (Kd = 4.8 μM); GSK-3β(WT) (IC50 = 515 nM) |
| ln Vitro | BRD0705 inhibits kinase function and does not stabilize β-catenin, mitigating potential neoplastic concerns. BRD0705 has no effect on healthy hematopoietic cells, but it causes myeloid differentiation and hinders colony formation in AML cells. [1] |
| ln Vivo | BRD0705 delays leukemia initiation and lengthens survival in AML mouse models.[1] |
| Cell Assay | After 24 hours of treatment with DMSO or BRD0705 at 20 μM, 30,000 to 50,000 cells are washed in PBS/2%FBS and then cytospun onto poly-L-Lysine coated slides (EMS). The cells are then blocked in 1% BSA/0.05% triton for 1 hour at room temperature, fixed in 4% paraformaldehyde (EMS) for 20 minutes, permeabilized in 0.2% Triton X-100 (RPI) for 30 minutes at 4°C, and incubated for 1 hour at room temperature with primary anti-β-catenin (1/500) and secondary antibody. After each antibody incubation, there are three 5 minute washes. The Yokogawa Spinning Disk Confocal / TIRF system is used to image the slides after mounting them with Prolong (R) Gold Antifade with DAPI. Image-J software is used to analyze representative images from three separate experiments. |
| Animal Protocol |
8-week-old male NSG mice injected with MLL-AF9 AML cells 30 mg/kg Oral gavage; twice daily |
| References |
[1]. Exploiting an Asp-Glu "switch" in glycogen synthase kinase 3 to design paralog-selective inhibitors for use in acute myeloid leukemia. Sci Transl Med. 2018 Mar 7;10(431). pii: eaam8460. |
Solubility Data
| Solubility (In Vitro) |
DMSO: 64~300 mg/mL (199.1~933.4 mM) Ethanol: ~64 mg/mL (~199.1 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 7.5 mg/mL (23.33 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 75.0 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: ≥ 7.5 mg/mL (23.33 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 75.0 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 | 2.8589 mL | 14.2943 mL | 28.5886 mL | |
| 5 mM | 0.5718 mL | 2.8589 mL | 5.7177 mL | |
| 10 mM | 0.2859 mL | 1.4294 mL | 2.8589 mL |