CRT0044876

製品コードS7449 バッチS744901

化学情報

	Synonyms	NSC 69877,7-NO2-ICA	Storage (From the date of receipt)	3 years -20°C powder 1 years -80°C in solvent
	化学式	C₉H₆N₂O₄
	分子量	206.15	CAS No.	6960-45-8
Solubility (25°C)*	体外	DMSO	41 mg/mL (198.88 mM)
		Ethanol	1 mg/mL (4.85 mM)
		Water	Insoluble
* <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.

溶剤液（一定の濃度）を調合する

生物活性

製品説明	CRT0044876 (NSC 69877, 7-NO2-ICA) is a potent and selective APE1 inhibitor with IC50 of ~3 μM.
in vitro	CRT0044876 potently inhibits the AP endonuclease, 3'-phosphodiesterase and 3'-phosphatase activities of APE1, and exhibits the selectivity for the inhibition of exonuclease III family. CRT0044876 potentiates the cytotoxicity of several DNA base-targeting compounds with an accumulation of unrepaired AP sites. ^[1] CRT0044876, by inhibition of the BER pathway increases oxidative DNA damage temporally related to increased intracellular reactive oxygen species in an acidic tumor microenvironment, and thus results in cell cycle arrests and increased DNA double strand breaks, leading to cell death. ^[2]

製品説明

CRT0044876 (NSC 69877, 7-NO2-ICA) is a potent and selective APE1 inhibitor with IC50 of ~3 μM.

in vitro

CRT0044876 potently inhibits the AP endonuclease, 3'-phosphodiesterase and 3'-phosphatase activities of APE1, and exhibits the selectivity for the inhibition of exonuclease III family. CRT0044876 potentiates the cytotoxicity of several DNA base-targeting compounds with an accumulation of unrepaired AP sites. ^[1] CRT0044876, by inhibition of the BER pathway increases oxidative DNA damage temporally related to increased intracellular reactive oxygen species in an acidic tumor microenvironment, and thus results in cell cycle arrests and increased DNA double strand breaks, leading to cell death. ^[2]

プロトコル(参考用のみ)

キナーゼアッセイ	AP site cleavage assay
キナーゼアッセイ	BER reaction buffer comprises 40 mM HEPES–KOH (pH 7.8), 5 mM MgCl₂, 0.5 mM DTT and 0.1 mM EDTA. A 10 μl AP site cleavage reaction comprised of BER buffer mix, purified protein (3.3 nM final concentration of APE1) and 0.75 ng reduced AP site double-stranded oligonucleotide. The mixture is incubated at 37°C for 1 h. A total of 1 μl of stop buffer (50% glycerol, 10 mM Tris–HCl, 1 mM EDTA, 0.1% bromophenol blue and 0.1% Xylene cyanol) is added, and the sample mixture is denatured at 90–100°C for 2 min. The sample is then run on a 15% TBE Criterion™ Pre-Cast Gel, with electrophoresis at a constant current of 30 mA for 30 min, and the radiolabeled substrate and reaction products are visualized using a phosphorImager. The inhibitory activity of potential APE1-targeting compounds are analyzed at drug concentrations ranging from 0.1 to 100 μM. The resolved radiolabeled bands are quantified using ImageQuant software analysis, and IC50 values are calculated by determining the concentration of the inhibitor that reduced APE1 activity to 50% of the control values.
細胞アッセイ	細胞株	HT1080 fibrosarcoma cells
	濃度	100-500 μM
	反応時間	7-10 days
	実験の流れ	HT1080 fibrosarcoma cells are grown in 2% RPMI medium [supplemented with penicillin 0.06 g/l, streptomycin 0.1 g/l (pH 7.0), 10% fetal bovine serum and 4 mM glutamine]. Only cells with a plating efficiency of ≥60% are used for clonogenic survival analysis. Tissue culture dishes (10 cm) are seeded with 500 cells per dish and the culture is maintained in a humidified incubator at 37°C in an atmosphere of 5% CO₂ and 95% air. To evaluate the toxicity profile of putative APE1 inhibitors, various concentrations (100–500 μM) of inhibitor are added to the medium, and cultures were incubated for 7-10 days until cell colonies are formed. Colonies are fixed [75% (v/v) methanol, 25% (v/v) acetic acid] for 30 min and stained with crystal violet (1 mg/ml in distilled water) for 4 h at room temperature. Visible colonies are counted on a colony counter.

参考

Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：

Synergistic drug combinations for cancer identified in a CRISPR screen for pairwise genetic interactions. [ Nat Biotechnol, 2017, 35(5):463-474]	PubMed: 28319085

Synergistic drug combinations for cancer identified in a CRISPR screen for pairwise genetic interactions. [ Nat Biotechnol, 2017, 35(5):463-474]

PubMed: 28319085

長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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