Fbxo3 is a ubiquitin E3 ligase F box component, which potently stimulates cytokine secretion from human inflammatory cells by mediating the degradation of the TRAF
inhibitory protein, Fbxl2. BC-1215 is a highly unique, selective Fbxo3 inhibitor with an estimated IC50 value of 10-7M, measured by prevention of SCFFbxo3 catalyzed Fbxl2 ubiquitination in vitro. BC-1215 exhibited maximal inhibitory binding at 10-7M, measured by quantification of bound Fbxl2 protein. As the inhibition of Fbxo3, the known Fbxl2 substrates including cyclin D2, cyclin D3, and cytidylyl-transferase were decreased in MLE cells treated with BC-1215 at 10μM and 50μM for 16h. The suppression of upregulated levels of TRAF2 and TRAF3, along with activating the downstream p38 and phospho-IKK in the NF-kB pathway, by TNF treatment could also be observed in MLE cells treated with 10μg/ml BC-1215 for 6h. BC-1215 reversibly inhibited Fbxo3 resulting in destabilized TRAF proteins. Intraperitoneal injection with BC-1215 at doses of 0.8, 4, 20, 100 and 500μg/kg, 10 minutes prior to LPS challenge, dose-dependently inhibited the release of IL-1b, IL-6, and TNF-α in blood of mice. In vivo administration of BC-1215 effectively lessened the severity of viral pneumonia, septic shock, colitis, and cytokine-driven inflammation systemically in murine models.
BC-1215 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Murine lung epithelial cells (MLE)
0, 0.4, 2, 10, 50 μg/mL
16 hours
BC-1215 decreased TRAF1-6 protein half-life from 8–12 h to 3–4 h, without altering TRAF steady-state mRNA levels.
J Immunol. 2013 Nov 15;191(10):5247-55.
Human peripheral blood mononuclear cells (PBMC)
0, 0.4, 2, 10, 50 μg/mL
16 hours
BC-1215 remarkably suppressed the majority of the Th1 panel cytokines including G-CSF, GM-CSF, GROα, I-309, IL1-α, IL1-β, IL1rα, IL-6, IL-12, IL-23, MIP-1α, MIP-1β and TNFα.
J Immunol. 2013 Nov 15;191(10):5247-55.
HT22 cells
10 μg/mL
2 hours
To confirm the pro-inflammatory effect of FBXO3 in the OGD/R model
Int J Mol Sci. 2022 Nov 7;23(21):13648.
A549 cells
10 μg/mL
24 hours hypoxia followed by 4 hours reoxygenation
BC-1215 significantly inhibited the H/R-induced increase in phospho-NF-κB p65 and decrease in IκB-α protein expression, and reduced IL-8 production.
Front Pharmacol. 2019 May 24;10:583.
BC-1215 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Sprague-Dawley rats
Lung ischemia-reperfusion injury model
Added to the perfusate
0.1, 0.25, 0.5 mg/kg
Single administration, observed for 100 minutes
BC-1215 significantly attenuated I/R-induced lung edema, inflammation, oxidative stress, and apoptosis, and inhibited the activation of NF-κB and MAPK signaling pathways.
Front Pharmacol. 2019 May 24;10:583.
Sprague-Dawley rats
Neuropathic pain model (spinal nerve ligation)
Intrathecal administration
10, 30, and 100 nM
Single bolus injection
BC-1215 ameliorated SNL-induced behavioral allodynia by antagonizing TRAF2/TNIK/GluR1 signaling
J Neurosci. 2015 Dec 16;35(50):16545-60
Sprague-Dawley rats
Spinal nerve ligation (SNL) model
Intrathecal injection
100 nM
Single bolus injection
BC-1215 attenuated SNL-induced allodynia by inhibiting Fbxo3 activity.
J Neurosci. 2016 Sep 14;36(37):9722-38
C57BL/6 mice
Sepsis model
Intraperitoneal injection
500 µg, 100 µg, 20 µg, 4 µg and 0.8 µg
Single dose
BC-1215 exhibited high potency in vivo (inhibitory dose [ID50] IL-1β = 1 mg/kg, ID50 IL-6 = 2.5 mg/kg, ID50 TNFα = 1.2 mg/kg).
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