Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Dec;39(1):2394895.
doi: 10.1080/14756366.2024.2394895. Epub 2024 Sep 2.

Expanding the inhibitor space of the WWP1 and WWP2 HECT E3 ligases

Ashley P Dudey  1 , Jake M Rigby  2 , Gregory R Hughes  1 , G Richard Stephenson  2 , Thomas E Storr  2 , Andrew Chantry  1 , Andrew M Hemmings  1   2   3
Affiliations

Expanding the inhibitor space of the WWP1 and WWP2 HECT E3 ligases

Ashley P Dudey et al. J Enzyme Inhib Med Chem. 2024 Dec.

Abstract

The HECT E3 ubiquitin ligases 1 (WWP1) and 2 (WWP2) are responsible for the ubiquitin-mediated degradation of key tumour suppressor proteins and are dysregulated in various cancers and diseases. Here we expand their limited inhibitor space by identification of NSC-217913 displaying a WWP1 IC50 of 158.3 µM (95% CI = 128.7, 195.1 µM). A structure-activity relationship by synthesis approach aided by molecular docking led to compound 11 which displayed increased potency with an IC50 of 32.7 µM (95% CI = 24.6, 44.3 µM) for WWP1 and 269.2 µM (95% CI = 209.4, 347.9 µM) for WWP2. Molecular docking yielded active site-bound poses suggesting that the heterocyclic imidazo[4,5-b]pyrazine scaffold undertakes a π-stacking interaction with the phenolic group of tyrosine, and the ethyl ester enables strong ion-dipole interactions. Given the therapeutic potential of WWP1 and WWP2, we propose that compound 11 may provide a basis for future lead compound development.

Keywords: SAR; WWP1; WWP2; drug discovery; ubiquitin ligase inhibitors.

PubMed Disclaimer

Conflict of interest statement

No potential conflict of interest was reported by the authors.

Figures

None
Graphical abstract
Figure 1.
Figure 1.
High throughput DSF screen of the NCI Diversity Set VI compound library against WWP1 and WWP2. Assays were carried out using 10 µM compound, containing 0.1% DMSO and normalised by ΔTm calculated from controls. Positive hits (green) displayed had an average ΔTm and associated errors above a threshold of three times the standard deviation of their respective controls (A) Single-shot DSF screen of WWP2-LH (top) and WWP1-L34H (bottom) against NCI Diversity Set VI compounds. (B) Triplicate DSF screen of 24 confirmed hit compounds for WWP1-L34H and WWP2-LH.
Figure 2.
Figure 2.
ELISA autoubiquitination assay screen of PAINS-filtered DSF hit compounds against WWP1 (A) and WWP2 (B). Enzyme inhibition was measured at a compound concentration of 500 µM (1% DMSO), with a hit threshold (hits shown in green) of less than 50% relative activity, normalised to their respective 0% and 100% WWP1-L34H and WWP2-FL controls.
Figure 3.
Figure 3.
Synthesised NSC-217913 against WWP1 and WWP2. (A) Chemical structure of NSC-217913 and associated WWP1 IC50 (B) Dose-dependent ELISA autoubiquitination assay of synthesised NSC-217913. Compound inhibition was measured on a log scale from 1000 – 10 nM (1% DMSO), normalised to 0% and 100% WWP1-L34H (blue triangle) and WWP2-FL (red square) controls. IC50 values were calculated from non-linear regression curves fitted in GraphPad.
Scheme 1.
Scheme 1.
Synthetic route to access 6, 7, 9, 11, 13 and 15. i: Ethylene glycol (2.04 M), 175 °C, 18 h, 45%. ii: PCl5 (7.0 eq), POCl3 (0.44 M), 90 °C – 200 °C, 18 h, 32%. iii: NH4OH (25% aq., 0.27 M), 120 °C, 2 d., 31%. iv: Thiocarbonyldiimidazole (2.0 eq), dioxane (0.2 M), 120 °C, 36 h, 34%. v: Br-R (1.05 eq), NaOH (1.5 eq), EtOH (0.067 M), RT, 18 h, Ar or N2, 24 – 76%. vi: NaOH (0.15 M, methanol), CH2Cl2: MeOH (14:11 v/v, 0.063 M), RT, 18 h, 69%.
Figure 4.
Figure 4.
2D and 3D ligand poses of compound 11 docked to (A) WWP1-2L34H and (B) WWP2-LH. Key interacting residues are represented as sticks and coloured associated with their hydrophilic (blue), hydrophobic (green) or positively (purple) and negative (orange) charged characteristics, with the binding pocket surface (grey) also shown. Interactions including π-π stacking (green), hydrogen bonding (pink), halogen bonding (orange) and electrostatic (red) are given in angstroms (Å). 2D and 3D images were created using the Maestro Schrodinger Suite and PYMOL, respectively.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Komander D, Rape M.. The ubiquitin code. Annu Rev Biochem. 2012;81(1):203–229. - PubMed
    1. Petroski MD. The ubiquitin system, disease, and drug discovery. BMC Biochem. 2008;9 (Suppl 1):S7. - PMC - PubMed
    1. Kane RC, Bross PF, Farrell AT, Pazdur R.. Velcade®: U.S. FDA approval for the treatment of multiple myeloma progressing on prior therapy. Oncologist. 2003;8(6):508–513. - PubMed
    1. Kwon YT, Ciechanover A.. The ubiquitin code in the ubiquitin-proteasome system and autophagy. Trends Biochem Sci. 2017;42(11):873–886. - PubMed
    1. Sharma A, Preuss CV. Bortezomib [Internet]. Treasure Island (FL): StatPearls Publishing; 2024. [cited 2024 Mar 21]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK519559.

MeSH terms

Grants and funding

The work was supported by funding to G.R.H. and A.C. from BigC Cancer Charity (Research Grant 19-14R) supporting a PhD studentship for J.M.R. A.P.D. was supported by a University of East Anglia (UEA) Science Faculty PhD Studentship.

LinkOut - more resources

  NODES
INTERN 3
twitter 2