Discovery of Potent and Selective 2-(Benzylthio)pyrimidine-based DCN1-UBC12 Inhibitors for Anticardiac Fibrotic Effects

Abstract

DCN1, a co-E3 ligase, interacts with UBC12 and activates cullin–RING ligases (CRLs) by catalyzing cullin neddylation. Although DCN1 has been recognized as an important therapeutic target for human diseases, its role in the cardiovascular area remains unknown. Here, we first found that DCN1 was upregulated in isolated cardiac fibroblasts (CFs) treated by angiotensin (Ang) II and in mouse hearts after pressure overload. Then, structure-based optimizations for DCN1-UBC12 inhibitors were performed based on our previous work, yielding compound DN-2. DN-2 specifically targeted DCN1 at molecular and cellular levels as shown by molecular modeling studies, HTRF, cellular thermal shift and co-immunoprecipitation assays. Importantly, DN-2 effectively reversed Ang II-induced cardiac fibroblast activation, which was associated with the inhibition of cullin 3 neddylation. Our findings indicate a potentially unrecognized role of DCN1 inhibition for anticardiac fibrotic effects. DN-2 may be used as a lead compound for further development.

Targeting cullin neddylation for cancer and fibrotic diseases

“…Moreover, given that the antioxidant transcription factor NRF2 is a typical substrate of cullin3 10 , DCN1 inhibitors probably play vital role for the treatment of diseases related to over-production of ROS. In particular, the recent findings disclosed that DCN1 inhibitors possessed evident potency on reducing acetaminophen-induced liver damage and AngⅡ-induced cardiac fibrosis, connected with the inhibition of cullin3 neddylation and its substrate NRF2 activation 142 , 145 . Besides, considering that neddylation inhibition well sensitize chemo-/radio-resistant cancer cells 159 , 167 , 172 , DCN1 inhibitors can be used to investigate their potency on relieving resistance in the future.…”

“…Our group observed that DCN1 protein was obviously upregulated in isolated cardiac fibroblasts treated by Angiotensin (Ang) II and in mouse hearts after pressure overload. However, inhibition of DCN1 effectively reversed Ang II-induced cardiac fibroblast activation, which was associated with the inhibition of cullin 3 neddylation and the accumulation of its substrate protein NRF2 145 . Besides, we also found that treated with DCN1 inhibitor could potentially relieved liver fibrosis in CCl4-induced mouse model 141 .…”

“…Based on the docking modes of DCN1 inhibitor 49 , we conducted further structural optimizations and SARs using several drug design strategies, including structure-based drug design and bioisosterism etc., yielding 77 novel 2-thiopyrimidine derivatives as DCN1 inhibitors 145 . In comparison to molecule 49 , the new synthesized compound 50 (IC 50 = 9.55 ± 0.98 nM) possessed stronger inhibitory effect, higher selectivity toward DCN-like proteins and weaker toxicity against cardiac fibroblasts (CFs) (Figure 23 A).…”

“…Considering the NRF2, an antioxidant transcription factor, is a typical substrate of cullin3 10 , DCN1 inhibitors could play extensive efficacy for the treatment of diseases linked to over-production of reactive oxygen species (ROS). Especially, the latest findings indicated that DCN1 inhibitors exhibited effective potency on relieving acetaminophen-induced liver damage and AngⅡ -induced cardiac fibrosis, which was related with the inhibition of cullin3 neddylation and its substrate NRF2 activation 142 , 145 . All in all, more potent and selective DCN1 inhibitors with drug-like properties are urgently required, aiming to further investigate their therapeutical potential for human diseases and DCN1 protein's potentially biological roles.…”

Identification of the communal pathogenesis and immune landscape between viral myocarditis and dilated cardiomyopathy

“…Its related pathways include Class I MHC‐mediated antigen processing and presentation and regulation of activated PAK‐2p34 by proteasome‐mediated degradation. DCN1 is a co‐E3 ligase that interacts with UBC12, and inhibitors of DCN1–UBC12 interaction are effective in alleviating myocardial fibrosis, 25 which is known to be associated with the prognosis of DCM. 26 The expression of UBC12 in chronic VMC samples was significantly different from normal samples, hence the indispensable role of UBC12 in improving the prognosis of DCM.…”