Home Epigenetics Histone Demethylase inhibitor GSK-J4 Hydrochloride

연구용

GSK-J4 Hydrochloride H3K27 Histone Demethylase Inhibitor

제품 번호: S7070

GSK J4 HCl is a cell permeable prodrug of GSK J1, which is the first selective inhibitor of the H3K27 histone demethylase JMJD3 and UTX with IC50 of 60 nM in a cell-free assay and inactive against a panel of demethylases of the JMJ family.
GSK-J4 Hydrochloride Histone Demethylase inhibitor Chemical Structure

화학 구조

분자량: 453.96

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품질 관리 (Quality Control)

배치: 순도: 99.48%
99.48

세포 배양, 처리 및 작업 농도
(Cell Culture, Treatment & Working Concentration)

세포주 분석 유형 농도 배양 시간 제형 활성 설명 PMID
CUTLL1 Growth inhibitory assay 2 μM DMSO affects cell growth 25132549
CUTLL1 Apoptosis assay 2 μM DMSO induces apoptosis 25132549
CUTLL1 Function assay 2 μM DMSO induces cell cycle arrest 25132549
CUTLL1 Kinase assay 6 μM DMSO leads to increased H3K27me3 25132549
SF7761 Kinase assay 6 μM DMSO increases K27 methylation 25401693
SF8628 Kinase assay 6 μM DMSO increases K28 methylation 25401693
H3.3 Kinase assay 6 μM DMSO increases K29 methylation 25401693
SF9012 Kinase assay 6 μM DMSO increases K30 methylation 25401693
SF9402 Kinase assay 6 μM DMSO increases K31 methylation 25401693
SF9427 Kinase assay 6 μM DMSO increases K32 methylation 25401693
human astrocytes Kinase assay 6 μM DMSO increases K33 methylation 25401693
SF7761 Growth inhibitory assay 6 μM DMSO inhibits K27M glioma cell growth 25401693
SF8628 Growth inhibitory assay 6 μM DMSO inhibits K28M glioma cell growth 25401693
H3.3 Growth inhibitory assay 6 μM DMSO inhibits K29M glioma cell growth 25401693
SF9012 Growth inhibitory assay 6 μM DMSO inhibits K30M glioma cell growth 25401693
SF9402 Growth inhibitory assay 6 μM DMSO inhibits K31M glioma cell growth 25401693
SF9427 Growth inhibitory assay 6 μM DMSO inhibits K32M glioma cell growth 25401693
human astrocytes Growth inhibitory assay 6 μM DMSO inhibits K33M glioma cell growth 25401693
TG neurons Function assay 50 μM DMSO inhibits HSV-1 reactivation from sensory neurons 25552720
Th17 Function assay 80 nM DMSO inhibits cell differentiation 25840993
β-cells Function assay 20 μM DMSO blunts IFNγ, Il-1β, and TNFα-induced chemokine gene expression 26505193
β-cells Function assay 20 μM DMSO induces β-cell dysfunction 26505193
ESCs Function assay 1.8 µM DMSO induces DNA damage along with activation of the DNA damage response 26759175
Raw 264.7 Function assay 0.8192 µM DMSO inhibits TNF-α production 26776360
MCF7 Function assay 1 to 10 uM 30 hrs Inhibition of KDM5A in human MCF7 cells assessed as effect on H3K4me3 methylation levels at 1 to 10 uM after 30 hrs by Western blot analysis 30392349
MCF7 Function assay 1 to 10 uM 30 hrs Inhibition of KDM5A in human MCF7 cells assessed as effect on H3K27me3 methylation levels at 1 to 10 uM after 30 hrs by Western blot analysis 30392349
RAW264.7 0.82 uM 24 hrs Inhibition of LPS-induced TNFalpha production in mouse RAW264.7 cells 0.82 uM after 24 hrs by ELISA 26776360
RAW264.7 24 hrs Inhibition of LPS-induced TNFalpha production in mouse RAW264.7 cells after 24 hrs by ELISA 26776360
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화학 정보, 보관 및 안정성 (Chemical Information, Storage & Stability)

분자량 453.96 화학식

C24H27N5O2.HCl

 보관 (수령일로부터)
CAS 번호 1797983-09-5 SDF 다운로드 원액 보관

용해도 (Solubility)

In vitro
배치:

DMSO : 91 mg/mL (200.45 mM)
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Ethanol : 91 mg/mL

Water : 10 mg/mL

몰농도 계산기

질량 농도 부피 분자량
희석 계산기 분자량 계산기

In vivo
배치:

생체 내 제형 계산기 (투명한 용액)

1단계: 아래 정보 입력 (권장: 실험 중 손실을 고려하여 추가 동물 포함)

mg/kg g μL

2단계: 생체 내 제형 입력 (이것은 계산기일 뿐 제형이 아닙니다. 용해도 섹션에 생체 내 제형이 없는 경우 먼저 당사에 문의하십시오.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

계산 결과:

작업 농도: mg/ml;

DMSO 원액 준비 방법: mg 약물 사전 용해 μL DMSO ( 원액 농도 mg/mL, 농도가 해당 약물 배치의 DMSO 용해도를 초과하는 경우 먼저 당사에 문의하십시오. )

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가μL PEG300, 혼합하고 투명하게 한 다음 추가μL Tween 80, 혼합하고 투명하게 한 다음 추가 μL ddH2O, 혼합하고 투명하게 합니다.

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가 μL 옥수수 기름, 혼합하고 투명하게 합니다.

참고: 1. 다음 용매를 추가하기 전에 액체가 투명한지 확인하십시오.
2. 용매를 순서대로 추가해야 합니다. 다음 용매를 추가하기 전에 이전 추가에서 얻은 용액이 투명한 용액인지 확인해야 합니다. 와동, 초음파 또는 뜨거운 물 중탕과 같은 물리적 방법을 사용하여 용해를 도울 수 있습니다.

작용 메커니즘 (Mechanism of Action)

Targets/IC50/Ki
JMJD3
(Cell-free assay)
60 nM
시험관 내(In vitro)

GSK J4 HCl is an ethyl ester derivative of the JMJD3 selective histone demethylase inhibitor GSK-J1 with an IC50 value greater than 50 μM in vitro. This compound is used to probe the consequences of demethylation of H3K27me3. In human primary macrophages, this chemical inhibits the lipopolysaccharide-induced production of cytokines, including pro-inflammatory tumour necrosis factor (TNF). In addition, it prevents the lipopolysaccharide-induced loss of H3K27me3 associated with the TNF transcription start sites and blocked the recruitment of RNA polymerase II.

키나아제 분석
Histone Demethylase AlphaScreen
Inhibition of histone demethylases is assessed using the histone demethylase AlphaScreen assay (Amplified Luminescence Proximity Homogenous Assay). This assay uses a biotinylated peptide substrate and relies on detection of the product methyl mark using a specific antibody coupled to protein-A acceptor beads and a Steptavidin donor bead to capture the peptide. In brief, recombinant demethylase enzymes are incubated in the presence of Fe2+ in the form of Ferrous Ammonium Sulphate (FAS), -ketoglutarate (KG) and biotinylated peptide substrate. L-Ascorbic Acid is included to provide a reducing environment and prevent oxidation of Fe2+. After incubation with peptide substrate the presence of the product is detected using AlphaScreen technology. The demethylase AlphaScreen assays are performed in 384-well plate format using white proxiplates. All steps are carried out in assay buffer (50 mM HEPES pH 7.5, 0.1% (w/v) BSA and 0.01 % (v/v) Tween-20). FAS is dissolved fresh each day in 20 mM HCl to a concentration of 400 mM and diluted to 1.0 mM in deionized water. All other components are dissolved fresh each day in deionized water. For IC50 determinations 5 μL of assay buffer containing demethylase enzyme is transferred to wells of a 384-well proxiplate. Titrations of this compound (0.1 μL) are transferred to each well and the enzymes allowed to pre-incubate for 15 minutes with this chemical (final concentration of DMSO is 1%). The enzyme reaction is initiated by addition of 5 μL of a substrate mix consisting of α-KG, FAS, L-Ascorbic Acid and biotinylated peptide substrate and the reaction incubated for the indicated time at room temperature. The enzyme reaction is stopped after the indicated time by addinton of 5 μL of EDTA (7.5 mM final concentration in assay buffer). Streptavidin Donor beads (0.08 mg/ml) and Protein-A conjugated acceptor beads (0.08 mg/ml) are pre-incubated for 1 hour with an antibody to the product methyl mark and the presence of biotin-H3-product is detected by addition of 5 μL of the preincubated AlphaScreen beads (final concentrations of 0.02 mg/ml with respect to acceptor and donor beads). Detection is allowed to proceed for 1 hour at room temperature and the assay plates read in a BMG Labtech Pherastar FS plate reader. Data are normalized to the no enzyme control and the IC50 determined from the nonlinear regression curve fit using GraphPad Prism 5.
생체 내(In vivo)

GSK-J4 hydrochloride is a potent dual inhibitor of H3K27me3/me2-demethylases JMJD3/KDM6B and UTX/KDM6A. It inhibits LPS-induced TNF-α production in human primary macrophages. This compound is a cell permeable prodrug of GSK-J1.
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