BIX 01294

Catalog No.S8006 Batch:S800602

Technical Data

Formula	C₂₈H₃₈N₆O₂.3HCl
Molecular Weight	600.02	CAS No.	1392399-03-9
Solubility (25°C)*	In vitro	DMSO	100 mg/mL (166.66 mM)
		Water	100 mg/mL (166.66 mM)
		Ethanol	100 mg/mL (166.66 mM)
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

BIX01294 is an inhibitor of G9a histone methyltransferase with IC50 of 2.7 μM in a cell-free assay, reduces H3K9me2 of bulk histones, also weakly inhibits GLP (primarily H3K9me3), no significant activity observed at other histone methyltransferases. BIX01294 induces autophagy. BIX01294 also inhibits H3K36 methylation by oncoproteins NSD1, NSD2 and NSD3.

Targets

G9a ^[1]
(Cell-free assay)

2.7 μM

In vitro

BIX01294 is a selective inhibitor of G9a histone methyltransferase and does not affect SUV39H1(H320R) and PRMT1 within the tested concentration range. BIX-01294 specifically inhibited G9a (H3K9me2) and, to a lesser extent, the closely related GLP enzyme (primarily H3K9me3), with an IC50 of 1.7 μM for G9a and 38 μM for GLP. BIX-01294 inhibits G9a in an uncompetitive manner with SAM. BIX-01294 (4.1μM) reduces H3K9me2 Levels in Bulk Histone Preparations from wt ES cells, mouse embryonic fibroblasts and HeLa cells, but not in G9a deficient stem cells. BIX-01294 is a valuable inhibitor for the transient modulation of chromosomal H3K9me2. BIX-01294 Reduces H3K9me2 at Several G9a Target Genes including Bim1 and Serac1. ^[1]

BIX01294 could reactivate expression of HIV-1 from latently infected cells such as ACH-2 and OM10.1. ^[2]

Protocol (from reference)

Kinase Assay:^{^[1]}	HMTase Assays Dissociation Enhanced Lanthanide Fluoro-ImmunoAssays (DELFIA) are performed in white, opaque 384-well plates coated with Neutravidin. Test compounds are diluted to 12 μg/ml in 50mM Tris-HCl pH 8.5containing 4% DMSO and 10 μl isdispensed into the wells. Blank and control wells received only compound buffer. GST-G9a at 10 μg/ml and SAM at 40 μM are 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 are 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 minutes. The plates are washed 3 times with 100 μl of Wash Buffer. Next, 50 μl of Fluoroimmunoassay (FI) Buffer containing 5ng α-2X-di-meth H3-K9 and 5ng goat anti-rabbit Eu chelate isadded to all wells of the plate, and the plate isincubated for an additional hour at room temperature. The plates are washed 3 times with 100 μL of Wash Buffer, and 50 μl of Enhancement Solution isadded to each well. Time resolved fluorescence ismeasured after 45 minutes on a Viewlux Microplate Imager imaging for 15 second.
Animal Study:^{^[3]}	Animal Models TetO-Her2/neu (TAN) mice Dosages 10 mg/kg Administration i.p.

Kinase Assay:^{^[1]}

HMTase Assays
Dissociation Enhanced Lanthanide Fluoro-ImmunoAssays (DELFIA) are performed in white, opaque 384-well plates coated with Neutravidin. Test compounds are diluted to 12 μg/ml in 50mM Tris-HCl pH 8.5containing 4% DMSO and 10 μl isdispensed into the wells. Blank and control wells received only compound buffer. GST-G9a at 10 μg/ml and SAM at 40 μM are 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 are 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 minutes. The plates are washed 3 times with 100 μl of Wash Buffer. Next, 50 μl of Fluoroimmunoassay (FI) Buffer containing 5ng α-2X-di-meth H3-K9 and 5ng goat anti-rabbit Eu chelate isadded to all wells of the plate, and the plate isincubated for an additional hour at room temperature. The plates are washed 3 times with 100 μL of Wash Buffer, and 50 μl of Enhancement Solution isadded to each well. Time resolved fluorescence ismeasured after 45 minutes on a Viewlux Microplate Imager imaging for 15 second.

Animal Study:^{^[3]}

Animal Models
TetO-Her2/neu (TAN) mice
Dosages
10 mg/kg
Administration
i.p.

References

Customer Product Validation

: Data from [Data independently produced by , , J Pineal Res, 2018, 65(2):e12497]

: Data from [Data independently produced by , , Cancer Res, 2015, 75(11):2375-86. ]

: Data from [Data independently produced by , , PLoS One, 2015, 10(9):e0138390]

Selleck's BIX 01294 has been cited by 47 publications

EHMT2 promotes tumorigenesis in GNAQ/11-mutant uveal melanoma via ARHGAP29-mediated RhoA pathway [ Acta Pharm Sin B, 2024, 14(3):1187-1203]	PubMed: 38486999
Gamma-glutamyl transferase secreted by Helicobacter pylori promotes the development of gastric cancer by affecting the energy metabolism and histone methylation status of gastric epithelial cells [ Cell Commun Signal, 2024, 22(1):402]	PubMed: 39148040
Extensive embryonic patterning without cellular differentiation primes the plant epidermis for efficient post-embryonic stomatal activities [ Dev Cell, 2023, 58(6):506-521.e5]	PubMed: 36931268
Depletion of G9A attenuates imiquimod-induced psoriatic dermatitis via targeting EDAR-NF-κB signaling in keratinocyte [ Cell Death Dis, 2023, 14(9):627]	PubMed: 37739945
Depletion of G9A attenuates imiquimod-induced psoriatic dermatitis via targeting EDAR-NF-κB signaling in keratinocyte [ Cell Death Dis, 2023, 14(9):627]	PubMed: 37739945
Perturbations in 3D genome organization can promote acquired drug resistance [ Cell Rep, 2023, 42(10):113124]	PubMed: 37733591
Restricting epigenetic activity promotes the reprogramming of transformed cells to pluripotency in a line-specific manner [ Cell Death Discov, 2023, 9(1):245]	PubMed: 37452056
Telomeres cooperate in zygotic genome activation by affecting DUX4/Dux transcription [ iScience, 2023, 26(3):106158]	PubMed: 36843839
The CHCHD2/Sirt1 corepressors involve in G9a-mediated regulation of RNase H1 expression to control R-loop [ Cell Insight, 2023, 2(4):100112]	PubMed: 37388553
The CHCHD2/Sirt1 corepressors involve in G9a-mediated regulation of RNase H1 expression to control R-loop [ Cell Insight, 2023, 2(4):100112]	PubMed: 37388553

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SHIPPING AND STORAGE
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