A potent small-molecule inhibitor of the DCN1-UBC12 interaction that selectively blocks cullin 3 neddylation

Zhou, Haibin; Lu, Jianfeng; Liu, Liu; Bernard, Denzil; Yang, Chao; Fernández‐Salas, Ester; Chinnaswamy, Krishnapriya; Layton, Stephanie Danielle; Stuckey, Jeanne A.; Yu, Qing; Zhou, Weihua; Pan, Zhen‐Qiang; Sun, Yi; Wang, Shaomeng

doi:10.1038/s41467-017-01243-7

Cited by 80 publications

(104 citation statements)

References 48 publications

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“…Following secondary antibody incubation, the membranes were developed by ECL (Advansta). Neddylation of CUL3 was detected by the presence of a double band at approximately 80 kDa (46,47). for 48 hours.…”

Section: Methodsmentioning

confidence: 99%

AGER1 downregulation associates with fibrosis in nonalcoholic steatohepatitis and type 2 diabetes

Dehnad

Fan

Jiang³

et al. 2020

Journal of Clinical Investigation

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Section: Methodsmentioning

confidence: 99%

AGER1 downregulation associates with fibrosis in nonalcoholic steatohepatitis and type 2 diabetes

Dehnad

Fan

Jiang³

et al. 2020

Journal of Clinical Investigation

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“…While, it was very stable from 43 to 61℃ in the 10µM DC-2 treated cells, similar to that treated by positive compound DI-591( Figure 9A and 9D). 30 In addition, DC-2 increases DCN1 thermal ability at concentration low to 1µM at 55℃ ( Figure 9B and 9E). These findings indicate that DC-2 can engage the cellular DCN1 protein and increase its thermal ability.…”

Section: Dc-2mentioning

confidence: 87%

“…52,55 In addition, the reported DCN1-UBE2M interaction inhibitors all have interactions with these three residues, either by forming hydrogen bonds or fitting into the hydrophobic pocket. 30,32,43 Therefore, we hypothesize that these three residues may play an important role in DCN1-UBE2M interaction. Furthermore, our above docking results also have confirmed that DC-2 has relatively strong interactions with Pro97, Phe164 and Tyr181 residues of DCN1 ( Figure 7A and 7B).…”

Section: Dc-2mentioning

confidence: 97%

“…However, the relative less cytotoxicity of DI-591 indicates its higher selectivity on DCN1-2 over DCN3-5. 30 Hz, ≡C-H). 13 Then, the cells were collected and measured by FACSCalibur flow cytometer (BD Biosciences).…”

Section: Dc-2mentioning

confidence: 98%

“…These data suggest that DC-2 has specificity and selectivity for DCN1 and 2, similar to the previously reported inhibitors (DI-591, NAcM-COV and DI-404). 30,32,43…”

Section: / 64mentioning

confidence: 99%

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Potent 5-Cyano-6-phenyl-pyrimidin-Based Derivatives Targeting DCN1–UBE2M Interaction

Zhou

Ding

et al. 2019

J. Med. Chem.

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Neddylation of the Cullin-RING E3 ligases (CRLs) regulates the homeostasis of approximately 20% of cellular proteins. Defective in cullin neddylation 1(DCN1), as a co-E3 ligase, interacts with UBE2M to enhance the activation of CRLs and this interaction is emerging as a therapeutic target for human diseases. Here, we present a series of pyrimidin-based small molecular inhibitors targeting DCN1-UBE2M interaction. After finding a novel inhibitor DC-1 with IC50 = 1.2µM, we performed a series of chemical optimization, which finally led to the discovery of a potent thiazole contained 5-cyano-6-phenylpyrimidin-based inhibitor DC-2 (IC50 = 15nM). Next, using protein and cellular thermal shift assays, co-immunoprecipitation, molecular docking and site specific mutation experiments, we further proved that DC-2 specifically inhibited the interaction of UBE2M and DCN1 at molecule and cellular levels, resulting in the decrease of cullin3 neddylation and accumulation of its substrate: NRF2. Our findings indicate that DC-2 may serve as a novel lead compound for specific derivatives targeting DCN1-UBE2M interaction. / 64  INTRODUCTION The ubiquitin-proteasome system (UPS) is integral to maintaining cellular protein homeostasis by regulating degradation of intracellular proteins. 1-7 The ubiquitylation pathway is executed by the coordinated efforts of the E1 (ubiquitin-activating enzyme), E2 (ubiquitinconjugating enzyme) and E3 (ubiquitin ligase enzyme) proteins. 5 Like ubiquitination, 8 neddylation is a novel type of posttranslational modification, in which the ubiquitin-like molecule NEDD8 is added to its target proteins and thus regulates their functions. 9, 10 Neddylation also consists of a tripartite enzymatic cascade, including the NEDD8-activating enzyme E1 (NAE, APPBP1 (NAE1) and UBA3 heterodimer), two NEDD8-conjugating enzymes E2s (UBE2F and UBE2M, also known as UBC12) and NEDD8-E3 ligases, 9, 10 with cullins being the best-characterized substrates. 11-22 Up to now, neddylation has been recognized to be highly activated in various cancers, 20, 27-33 which draws much attraction for oncologic drug discovery, especially on the finding of NEDD8 E1 inhibitor Pevonedistat(MLN4924). 10, 23-26 MLN4924 covalently binds to NEDD8 E1, blocking all CRLs neddylation, 7 thus causing accumulation of CRL substrates. 7, 10 Since MLN4924 possesses immense anti-cancer effects both in vitro and in vivo, 10, 27, 28-43 it has been approved for Phase II clinical trials for treatment of human acute myeloid leukemia, non-small cell lung cancer and mesothelioma. 29 However, as a result of its broad ablation of neddylation, MLN4924 has a series of toxicities. Therefore, alternative targeting specific CRLs may be more potential and safer for cancer treatment. 30-32 DCN1 (defective in cullin neddylation 1), also called DCUN1D1, DCNL1 or SCCRO, is a 4 / 64 highly conserved gene and amplified along the 3q26.3 in most squamous cell carcinomas and some other human cancers. 33-39 As a co-E3 ligase, it binds to the activation complex of Cullin-RBX1-UBE2M...

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Neddylation‐CRLs regulate the functions of Treg immune cells

Sun

2023

BioEssays

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Neddylation, a ubiquitylation-like post-translational modification, is catalyzed by a cascade composed of three enzymes: E1 activating enzyme, E2 conjugating enzyme, and E3 ligase with cullins as physiological substrates. Specifically, neddylation E2 UBE2M couples with E3 RBX1 to neddylate cullins 1-4, whereas neddylation E2 UBE2F couples with E3 RBX2/SAG to neddylate cullin 5, leading to activation of CRL1-4 (Cullin-RING ligases 1-4) and CRL5, respectively. While over-activation of the neddylation-CRLs axis occurs frequently in many human cancers, how neddylation-CRLs regulate the function of immune cells, particularly Treg cells was previously unknown. To this end, we recently performed Treg selective knockout of two neddylation E2s and two E3s, individually, driven by Foxp3-Cre, and found that while the Ube2f-Sag E2/E3 pair plays a minimal role, if any, the Ube2m-Rbx1 pair is essential for the maintenance of Treg functionality, since their deletion triggers robust inflammatory response with autoimmune phenotypes. Milder phenotype severity upon Treg KO of upstream Ube2m than that of downstream Rbx1 strongly suggested that Rbx1 regulates Treg function in a manner dependent and independent of neddylation.

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A potent small-molecule inhibitor of the DCN1-UBC12 interaction that selectively blocks cullin 3 neddylation

Cited by 80 publications

References 48 publications

AGER1 downregulation associates with fibrosis in nonalcoholic steatohepatitis and type 2 diabetes

AGER1 downregulation associates with fibrosis in nonalcoholic steatohepatitis and type 2 diabetes

Potent 5-Cyano-6-phenyl-pyrimidin-Based Derivatives Targeting DCN1–UBE2M Interaction

Neddylation‐CRLs regulate the functions of Treg immune cells

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