Targeting NEDD8-activating enzyme for cancer therapy - developments, clinical trials, challenges and future research directions

Sep 24, 2026

NEDDylation, a post-translational modification driven by a three-step enzymatic cascade, regulates diverse biological processes, with NEDD8-activating enzyme (NAE) acting as the sole activation enzyme. This review focuses on NAE as a cancer-drug target, summarizing progress in NAE inhibitors, including covalent agents such as MLN4924 and TAS4464 entering cancer clinical trials. It explains links between NEDDylation and cancers, NAE structural features, multistep NEDD8 activation mechanisms, and the pharmacological basis of clinically evaluated inhibitors, while analyzing key therapeutic challenges and future research directions.

LanguageEnglish

CategoryHealthcare

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Pages25

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Abstract

Background

NEDD8 and the NEDDylation cascade

NAE activation and downstream enzymes

NEDDylation substrates and CRLs

Therapeutic rationale: targeting NAE

MLN4924 and anti-cancer strategy

Fu and Wang Journal of Hematology & Oncology (2023) 16:87 [https://doi.org/10.1186/s13045-023-01485-7](https://doi.org/10.1186/s13045-023-01485-7) Journal of Hematology & Oncology REVIEW Open Access Targeting NEDD8-activating enzyme for cancer therapy: developments, clinical trials, challenges and future research directions Dong‑Jun Fu 1 and Ting Wang1* Abstract NEDDylation, a post‑translational modification through three‑step enzymatic cascades, plays crucial roles in the regu‑ lation of diverse biological processes. NEDD8‑activating enzyme (NAE) as the only activation enzyme in the NEDDyla‑ tion modification has become an attractive target to develop anticancer drugs. To date, numerous inhibitors or ago‑ nists targeting NAE have been developed. Among them, covalent NAE inhibitors such as MLN4924 and TAS4464 currently entered into clinical trials for cancer therapy, particularly for hematological tumors. This review explains the relationships between NEDDylation and cancers, structural characteristics of NAE and multistep mechanisms of NEDD8 activation by NAE. In addition, the potential approaches to discover NAE inhibitors and detailed pharmaco‑ logical mechanisms of NAE inhibitors in the clinical stage are explored in depth. Importantly, we reasonably investi‑ gate the challenges of NAE inhibitors for cancer therapy and possible development directions of NAE‑targeting drugs in the future. Keywords NEDDylation, NAE, Cancer therapy, Challenges, Development directions Background Neuronal precursor cell-expressed developmentally down-regulated protein 8 (NEDD8) shares about 60% of the amino acids with ubiquitin, which covalently binds to substrate proteins by generating an isopeptide chain between the lysine residue of substrates and the glycine residue of NEDD8 [1–5]. NEDDylation is a biochemical process of post-translational modification that conjugates NEDD8 to substrate proteins through the successive enzymatic cascades [6–10]. In the initial stage of NEDDylation, the precursor of NEDD8 is hydrolyzed to mature NEDD8 by the precursor processing enzymes [11–14]. Next, the mature NEDD8 is activated in the *Correspondence: Ting Wang [wangting1973@sina.com](wangting1973@sina.com) 1 Beijing Research Institute of Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China presence of adenosine triphosphate (ATP) by the E1 NEDD8-activating enzyme (NAE) consisting of amyloid protein-binding protein 1 (APPBP1) and ubiquitin-like modifier activating enzyme 3 (UBA3) [15–17]. Then, the activated NEDD8 is transferred to NEDD8-conjugating enzyme E2s (UBC12 and UBE2F) via a trans-thiolation process [18–23]. Finally, NEDD8 is transferred from NEDD8-conjugating enzyme E2s to a specific lysine residue of substrates through the catalytic action of NEDD8 E3 ligases [24–26]. In recent years, homogeneous timeresolved fluorescence [27], AlphaScreen [28], in vitro NEDD8 conjugation assay [29], cellular thermal shift assay [30], co-immunoprecipitation [31], pull-down assay [32] and ATP kinetics assay [33] are commonly used to identify NEDDylation process. The main substrates for NEDDylation pathway are members of the cullin family (cullin1, 2, 3, 4A, 4B and 5), which are the core components of Cullin-RING ligases (CRLs) [34–36]. The activation of CRLs requires NEDD8 © The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use

FAQs

Why is NEDD8-activating enzyme (NAE) considered an attractive target for anticancer drug development?

NAE is the only activation enzyme in the NEDDylation modification cascade. Blocking NAE can disrupt NEDDylation-dependent regulation that supports cancer-relevant biological processes.

Which covalent NAE inhibitors have reached clinical trials for cancer therapy?

Covalent NAE inhibitors such as MLN4924 and TAS4464 have entered clinical trials, particularly for hematological tumors.

What are the key mechanistic steps involved in NEDD8 activation by NAE?

NEDD8 is first matured by precursor processing enzymes, then activated in an ATP-dependent manner by the E1 NAE complex, transferred to E2 conjugating enzymes, and finally delivered to substrate lysine residues via NEDD8 E3 ligases.

Sep 24, 2026

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