A Sesquiterpene Lactone from a Medicinal Herb Inhibits Proinflammatory Activity of TNF-α by Inhibiting Ubiquitin-Conjugating Enzyme UbcH5

Abstract

UbcH5 is the key ubiquitin-conjugating enzyme catalyzing ubiquitination during TNF-α-triggered NF-κB activation. Here, we identified an herb-derived sesquiterpene lactone compound IJ-5 as a preferential inhibitor of UbcH5 and explored its therapeutic value in inflammatory and autoimmune disease models. IJ-5 suppresses TNF-α-induced NF-κB activation and inflammatory gene transcription by inhibiting the ubiquitination of receptor-interacting protein 1 and NF-κB essential modifier, which is essential to IκB kinase activation. Mechanistic investigations revealed that IJ-5 preferentially binds to and inactivates UbcH5 by forming a covalent adduct with its active site cysteine and thereby preventing ubiquitin conjugation to UbcH5. In preclinical models, pretreatment of IJ-5 exhibited potent anti-inflammatory activity against TNF-α- and D-galactosamine-induced hepatitis and collagen-induced arthritis. These findings highlight the potential of UbcH5 as a therapeutic target for anti-TNF-α interventions and provide an interesting lead compound for the development of new anti-inflammation agents.

Structural analysis of recombinant human ubiquitin-conjugating enzyme UbcH5c

“…Recently, it was reported that IJ-5 directly binds to and inactivates UbcH5c in cells, and exerts anti-inflammatory effects in vivo , which further validates UbcH5c as a therapeutic target for anti-TNF- α interventions 16 . Therefore, development and discovery of UbcH5c inhibitors will provide new anti-inflammatory agents.…”

“…Consequently, UbcH5 plays an important role in TNF- α –triggered NF- κ B pathway. According to the previous report, IJ-5 was discovered as a potential inhibitor of UbcH5c, which exhibited anti-inflammatory activity against TNF- α – and D-galactosamine–induced hepatitis and collagen-induced arthritis 16 . Therefore, UbcH5c may serve as a potent therapeutic target for the treatment of inflammatory and autoimmune diseases induced by aberrant activation of NF- κ B.…”

Elucidating Direct Kinase Targets of Compound Danshen Dropping Pills Employing Archived Data and Prediction Models

“…The drug molecules are connected to biocompatible inert resin (magnetic beads or agarose gel) through chemical reaction. The target protein of the drug molecule was determined by electrophoresis, purification and high resolution mass spectrometry [42][43][44][45][46]. Using this strategy, a series of targets for active components of TCM have been successfully identified.…”

Dual androgen receptor (AR) and STAT3 inhibition by a compound targeting the AR amino‐terminal domain

“…LNCaP cells were transfected with 200 μg/T75 flask FLAG-STAT3 and 200 μg/T150 flask FLAG-AR-NTD plasmids for 24 hours, followed by 1 μmol/L 154, 10 μmol/L Crypt with or without 50 ng/mL IL6 for 6 hours. LNCaP cells were collected in the medium and centrifuged at 800g for 5 minutes, then washed with cold PBS twice before adding into the cells soft RIPA buffer (PBS, 1% sodium deoxycholate, 20 mmol/L sodium molybdate, 50 mmol/L NaF, 25…”