E6446 serves as a potent, orally active antagonist of TLR7 and TLR9, employed in studying harmful inflammatory responses. It is also a strong SCD1 inhibitor (KD: 4.61 μM), markedly reducing adipogenic differentiation and hepatic lipogenesis via SCD1-ATF3 signaling. Furthermore, E6446 ameliorates liver pathology in mice fed a high-fat diet (HFD), suggesting its utility in researching non-alcoholic fatty liver disease (NAFLD)[1][2][3].
体内研究
E6446 (20 mg/kg, pO.) almost entirely inhibits the production of IL-6 induced by CpG1668, and in a dose-dependent manner, diminishes the development of ANA (anti-nuclear antibodies) in mice at levels of 20 and 60 mg/kg[1].
E6446 (20, 60 mg/kg, pO.) inhibits TLR9 signaling in mice in a dose-dependent fashion. At doses of 60, 120 mg/kg, orally, it prevents TLR and LPS-induced septic shock hyperresponsiveness in rodent malaria, reduces TLR responsiveness in acute malaria, and inhibits activation of both TLR7 and TLR9[2].
体外研究
E6446 is a powerful and orally active inhibitor of TLR7 and TLR9. It effectively inhibits DNA activation of HEK:TLR9 cells with an IC50 of 10 nM, yet its efficacy is significantly reduced in suppressing LPS endotoxin activation of HEK:TLR4 cells or R848 activation of HEK:TLR7 cells. E6446 strongly reduces IL-6 production triggered by CpG2216 but does not affect IL-6 production induced by the TLR3 ligand poly inosine-cytosine. The inhibitory effect of E6446 on TLR7 varies with the ligand; it is a strong inhibitor of IL-6 production induced by RNA but less effective against IL-6 induction by the small molecule imidazoquinoline ligand R-848. E6446 inhibits the TLR9-DNA interaction in vitro, with an IC50 ranging from 1 to 10 µM[1].
E6446 (0.01-0.03 μM) specifically blocks TLR9 activation by CpG ODN 2006, and inhibits TLR7/8 activation by the imidazoquinoline compound R848 at concentrations of 2-8 μM. It diminishes TLR4 activation by 50% at 30 μM, exhibiting IC50 values of 0.01 μM in HEK-TLR9 cells activated with oligo 2006 and 0.23 μM in human PBMCs activated with oligo 2216, respectively[2].
E6446 细胞实验
Cell Line
Concentration
Treated Time
Description
References
OP9 cells
10 µM
15 days
E6446 significantly inhibited adipogenic differentiation, reduced lipid droplet accumulation, and downregulated the expression of adipocyte differentiation marker genes Pparg, Fasn, Fabp4, and C/ebpα.
Cell Commun Signal. 2023 Sep 30;21(1):268.
AML12 cells
10 µM
48 hours
E6446 significantly inhibited hepatic steatosis, reduced lipid droplet accumulation, and downregulated the expression of lipogenesis-related genes C/ebpα, Fasn, and Fabp4.
Cell Commun Signal. 2023 Sep 30;21(1):268.
Cardiomyocytes
10 µM
6 hours
E6446 significantly reduced Il6 and Il1b mRNA expression in CCCP-treated cardiomyocytes but had no effect on the number of PicoGreen and LC3B double-positive deposits.
JACC Basic Transl Sci. 2019 May 22;4(3):348-363.
Cardiomyocytes
0 to 10 µM
6 hours
E6446 significantly reduced the induction of Il6, Il1b, and Tnfa mRNAs in response to ODN1668 but had no effect on the cytokine mRNAs induced by LPS or loxoribine.
JACC Basic Transl Sci. 2019 May 22;4(3):348-363.
Mouse spleen cells
0.01–0.1 µM
72 hours
E6446 diminished IL-6 production in response to TLR9 agonist
Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3689-94.
Human peripheral blood mononuclear cells (PBMCs)
0.05 µM
72 hours
E6446 diminished IL-6 production in response to TLR9 and TLR8 agonists
Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3689-94.
HEK293 cells
0.01–0.03 µM
Overnight
E6446 specifically inhibited TLR9 activation
Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3689-94.
E6446 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6J mice
Pressure overload-induced heart failure model
Oral
1.5 mg/200μL
Every 2 days for 4 weeks
E6446 significantly attenuated TAC-induced left ventricular dilatation and cardiac dysfunction, reduced cardiac fibrosis and inflammatory responses.
JACC Basic Transl Sci. 2019 May 22;4(3):348-363.
Sprague-Dawley rats
Monocrotaline-induced pulmonary hypertension model
Oral administration in drinking water
10 mg/kg
From 3 days before to 21 days after monocrotaline injection
To evaluate the preventive and therapeutic effects of TLR9 inhibitors on pulmonary hypertension, results showed that E6446 significantly improved hemodynamic parameters, vascular remodeling, and survival in pulmonary hypertension
J Am Heart Assoc. 2021 Apr 6;10(7):e019247
C57BL/6 mice
Experimental cerebral malaria (ECM) model
Oral
120 mg/kg
Once daily from day −1 to day 12 post-infection
E6446 treatment reduced proinflammatory cytokine production and prevented the development of ECM
Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3689-94.
C57BL/6J mice
High-fat diet-induced nonalcoholic fatty liver disease model
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