Journal article
FUNCTIONAL ANALYSIS OF BIPARTITE NRF2 ACTIVATORS THAT OVERCOME FEEDBACK REGULATION FOR AGE-RELATED CHRONIC DISEASES
Redox biology, v 86, 103794
30 Jul 2025
PMID: 40779837
Abstract
Activating Nrf2 with small molecules is a promising strategy for countering aging, oxidative stress, inflammation, and various disorders, including neurodegeneration. The primary regulator of Nrf2 protein stability is Keap1, a redox sensor protein and an adapter in the Cullin III ubiquitin ligase complex, which labels Nrf2 for proteasomal degradation. The canonical Nrf2 activators either chemically modify sensor thiols in Keap1 or competitively displace Nrf2 from the ubiquitin ligase complex. The latter approach is considered the most suitable for continuous administration, as non-specific chemical modifiers of Keap1 thiols also modify active thiols on other cellular proteins, causing side effects. However, when transitioning from homogeneous cell-free to cell-based assays, genuine displacement activators show a significant loss in potency by several orders of magnitude. We demonstrate that this discrepancy arises due to higher micromolar concentrations of Keap1 in cell lines. The absolute amounts of Nrf2 and Keap1 determined in brain sub-regions show more than an order of magnitude excess of Keap1 over Nrf2. A potential solution could involve targeted delivery of an alkylating agent to Keap1 to achieve the desired specificity. Transcriptomic analysis of a cell-permeable Nrf2 peptide bearing an alkylating fumarate moiety indicates selective activation of the Nrf2 genetic program, confirming the high specificity of this approach. Activation of the Nrf2-genetic program has a built-in feedback regulatory mechanism through Bach1, an Nrf2 transcriptional repressor, whose levels are elevated in age-related neurodegeneration. Thus, a benign bipartite Nrf2 activator with Bach1 inhibition properties is needed for maximal benefits. The recently developed heterocyclic carboxamide, HPPE, exhibits overlap with the Nrf2 pathway activated by the fumarate-linked Nrf2 peptide, an Nrf2 activator, as well as with zinc and tin protoporphyrins, which are inhibitors of Bach1. Therefore, HPPE presents a promising and unique combination of the two desired activities that could be further optimized to treat age-related neurodegeneration.
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•A high intracellular concentration of Keap1 is the primary reason for a decreased potency of reversible displacement activators of Nrf2 in cell cultures and in vivo.•Cell-permeable fumarate-linked Nrf2 peptide specifically activates the Nrf2-genetic program through targeted alkylation of Keap1.•HPPE, a small bipartite molecule, exhibits properties of both Nrf2 activation and Bach1 inhibition, to bypass the negative feedback regulation.
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Details
- Title
- FUNCTIONAL ANALYSIS OF BIPARTITE NRF2 ACTIVATORS THAT OVERCOME FEEDBACK REGULATION FOR AGE-RELATED CHRONIC DISEASES
- Creators
- Dmitry M. Hushpulian - Federal Center Research Fundamentals of BiotechnologyNavneet Ammal Kaidery - Medical University of South CarolinaPriyanka Soni - Darby Children’s Research InstituteAndrey A. Poloznikov - SciBerg e.Kfm, James-Monroe-Ring 107, Mannheim, GermanyArpenik A. Zakhariants - Institute of Bioorganic ChemistryAlexandra V. Razumovskaya - National Research University Higher School of EconomicsMariia O. Silkina - National Research University Higher School of EconomicsVladimir I. Tishkov - Lomonosov Moscow State UniversityEliot H. Kazakov - New York Medical CollegeAbraham M. Brown - New York Medical CollegeIrina N. Gaisina - University of Illinois Urbana-ChampaignYoung-Hoon Ahn - Drexel UniversitySergey V. Kazakov - Pace UniversityNancy Krucher - Pace UniversitySudarshana M. Sharma - MUSC Hollings Cancer CenterBindu D. Paul - Johns Hopkins MedicineIrina G. Gazaryan - Pace UniversitySergey V. Nikulin - Faculty of Biology and Biotechnologies, HSE University, Moscow, RussiaBobby Thomas - Medical University of South Carolina
- Publication Details
- Redox biology, v 86, 103794
- Publisher
- Elsevier
- Number of pages
- 15
- Grant note
- National Institutes of Health: P01CA203653, R01AG077396, R01NS101967, R01NS133688 Department of Defense: HT94252310443 Paul Allen Foundation: 19PABH134580006, R01AG071512, 1R21AG073684 Solve-ME FoundationCatalyst Award from Johns Hopkins UniversityAmerican Heart Association
The work was supported by grants funded by the National Institutes of Health (R01AG077396, R01NS101967, and R01NS133688) and the Department of Defense (HT94252310443) to BT. This work was supported by the Basic Research Program at the National Research University Higher School of Economics to SN, AR, and MS for computer modeling and bioinformatic analysis. SMS is partially supported by the National Institutes of Health grant P01CA203653. BDP is supported by the American Heart Association and Paul Allen Foundation grant 19PABH134580006, R01AG071512, 1R21AG073684, the Solve-ME Foundation, and the Catalyst Award from Johns Hopkins University.
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Chemistry
- Web of Science ID
- WOS:001547467000001
- Scopus ID
- 2-s2.0-105012430905
- Other Identifier
- 991022076208904721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology