BIX 01294
BIX-01294 is a small molecule inhibitor of G9a and GLP (a H3K9 PKMT that shares 80% sequence identity with G9a in their respective SET domains) that was discovered via screening of a library of 125,000 synthetic compounds. BIX-01294 is selective for G9a and GLP over several H3K9 PKMTs including SUV39H1 and ESET, other KMTs such as SET7/9, and the arginine methyltransferase PRTM1. The X-ray crystal structure of GLP and BIX-01294 confirmed that BIX-01294 bound to the histone peptide binding pocket but failed to interact with the lysine binding channel. Cells dosed with BIX-01294 at 4.1 μM were characterized by reduced H3K9me2 levels in several cell lines but toxicity to cells at > 4.1 μM was observed.
Reference
William P. Janzen, Tim J. Wigle, Jian Jin, Stephen V. Frye. Epigenetics: tools and technologies. Drug Discov Today Technol. 2010 ; 7(1): 59 – 65.
- 1. Yu-Rong Mu, Sheng-Yu Zou, et al. "Role and mechanism of FOXG1-related epigenetic modifications in cisplatin-induced hair cell damage." Front Mol Neurosci. 2023 Apr 26:16:1064579. PMID: 37181652
- 2. Agnieszka Grabowska, Hanna Sas-Nowosielska, et al. "Activation-induced chromatin reorganization in neurons depends on HDAC1 activity." Cell Rep. 2022 Feb 15;38(7):110352. PMID: 35172152
| Physical Appearance | A solid |
| Storage | Store at -20°C |
| M.Wt | 490.65 |
| Cas No. | 935693-62-2 |
| Formula | C28H38N6O2 |
| Solubility | ≥24.45 mg/mL in DMSO; ≥5.1 mg/mL in EtOH with ultrasonic; ≥51.2 mg/mL in H2O |
| Chemical Name | N-(1-benzylpiperidin-4-yl)-6,7-dimethoxy-2-(4-methyl-1,4-diazepan-1-yl)quinazolin-4-amine |
| SDF | Download SDF |
| Canonical SMILES | CN1CCCN(CC1)C2=NC3=CC(=C(C=C3C(=N2)NC4CCN(CC4)CC5=CC=CC=C5)OC)OC |
| Shipping Condition | Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice. |
| General tips | We do not recommend long-term storage for the solution, please use it up soon. |
| Kinase experiment [1]: | |
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HMTase Assays |
Dissociation Enhanced Lanthanide Fluoro-Immuno Assay (DELFIA) was performed in white, opaque 384-well plates coated with Neutravidin. Test compounds were diluted to 12 μg/mL in 50mM Tris-HCl (pH 8.5) containing 4% DMSO and 10 μL was dispensed into the wells. Blank and control wells received only compound buffer. GST-G9a at 10 μg/mL and SAM at 40 μM were diluted in 50mM Tris HCl (pH 8.5)/10 mM DTT and added in a volume of 20 μL. Blank wells received Tris/DTT buffer only. The reactions were initiated by the addition of 800 nM H3(1-20)-cysbiotin substrate in 50 mM Tris (pH 8.5) in a volume of 10 μL, and incubated at room temperature for 60 mins. The plates were washed 3 times with 100 μL of Wash Buffer. Next, 50 μL of Fluoroimmunoassay (FI) Buffer containing 5 ng α-2X-di-meth H3-K9 and 5 ng goat anti-rabbit Eu chelate was added to all wells of the plate, and the plate was incubated for an additional 1 hr at room temperature. The plates were washed 3 times with 100 μL of Wash Buffer, and 50 μL of Enhancement Solution was added to each well. Time resolved fluorescence was measured after 45 mins on a Viewlux Microplate Imager imaging for 15 seconds. |
| Cell experiment [1]: | |
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Cell lines |
Wild-type and G9a deficient mouse embryonic stem (ES) cells, mouse embryonic fibroblasts and human HeLa cells |
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Preparation method |
The solubility of this compound in DMSO is > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
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Reaction Conditions |
4.1 μM; over 2 days |
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Applications |
BIX-01294 (4.1 μM) reduced the H3K9me2 Levels in Bulk Histone Preparations from wild-type mouse ES cells, mouse embryonic fibroblasts and human HeLa cells, but not in G9a deficient mouse ES cells. |
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References: [1]. Kubicek S, O'Sullivan RJ, August EM, Hickey ER, Zhang Q, Teodoro ML, Rea S, Mechtler K, Kowalski JA, Homon CA, Kelly TA, Jenuwein T. Reversal of H3K9me2 by a small-molecule inhibitor for the G9a histone methyltransferase. Mol Cell. 2007 Feb 9;25(3):473-81. |
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| Description | BIX 01294 is an inhibitor of G9a-like protein and G9a histone lysine methyltransferase with IC50 value of 0.7 and 1.7 μM, respectively. | |||||
| Targets | G9a-like protein | G9a histone lysine methyltransferase | ||||
| IC50 | 0.7 μM | 1.7 μM | ||||
Quality Control & MSDS
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Chemical structure
