BI-8622 is a novel, potent and specific inhibitor of the ubiquitin ligase HUWE1 with an IC50 of 3.1 μM. Deregulated expression of MYC is a driver of colorectal carcinogenesis, necessitating novel strategies to inhibit MYC function. The ubiquitin ligase HUWE1 (HECTH9, ARF-BP1, MULE) associates with both MYC and the MYC-associated protein MIZ1.
Physicochemical Properties
| Molecular Formula | C25H26N6O |
| Molecular Weight | 426.513544559479 |
| Exact Mass | 426.216 |
| CAS # | 1875036-74-0 |
| PubChem CID | 138377590 |
| Appearance | White to off-white solid powder |
| LogP | 2.7 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 6 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 32 |
| Complexity | 667 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O=C(C1=C(C)C(=NC=N1)N1CCC(C#N)(C2C=CC=CC=2)CC1)NC1C=CC(CN)=CC=1 |
| InChi Key | IJHAXMJRQQTBPL-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C25H26N6O/c1-18-22(24(32)30-21-9-7-19(15-26)8-10-21)28-17-29-23(18)31-13-11-25(16-27,12-14-31)20-5-3-2-4-6-20/h2-10,17H,11-15,26H2,1H3,(H,30,32) |
| Chemical Name | N-(4-(Aminomethyl)phenyl)-6-(4-cyano-4-phenylpiperidin-1-yl)-5-methylpyrimidine-4-carboxamide |
| Synonyms | BI8622; BI 8622; BI-8622 |
| 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
| ln Vitro | BI8622 has an IC50 value of 6.8 μM and causes ectopic production of HUWE1 in HeLa cells, which eliminates ubiquitination of MCL1 [1]. With an approximate IC50 value of 8.4 μM, BI8622 suppresses the development of Ls174T cell colonies[1]. All phases of the cell cycle are delayed in Ls174T cells when treated with BI8622 (10 μM; 1-4 days), with the greatest effect occurring in G1 [1]. BI8622 (0-50 μM; 16 hours) inhibits HUWE1 in HeLa cells, delaying MCL1 degradation in response to UV exposure [1]. Colorectal cancer cells' MYC-dependent transactivation is inhibited by BI8622 [1]. |
| Cell Assay |
Cell Cycle Analysis[1] Cell Types: Ls174T Cell Tested Concentrations: 0 μM, 5 μM, 10 μM, 15 μM, 20 μM Incubation Duration: 0-4 Days Experimental Results: Passaging of Ls174T cells was delayed in all phases of the cell cycle, G1 has the strongest effect. Western Blot Analysis [1] Cell Types: HeLa cells Tested Concentrations: 0 μM, 10 μM, 20 μM Incubation Duration: 16 hrs (hours) Experimental Results: Delaying the degradation of MCL1 in response to UV irradiation by inhibiting HUWE1 in HeLa cells. |
| References |
[1]. Tumor cell-specific inhibition of MYC function using small molecule inhibitors of the HUWE1 ubiquitin ligase. EMBO Mol Med. 2014 Dec;6(12):1525-41. |
Solubility Data
| Solubility (In Vitro) | DMSO : ~125 mg/mL (~293.08 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.88 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 (4.88 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 (4.88 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 | 2.3446 mL | 11.7231 mL | 23.4461 mL | |
| 5 mM | 0.4689 mL | 2.3446 mL | 4.6892 mL | |
| 10 mM | 0.2345 mL | 1.1723 mL | 2.3446 mL |