Modulating receptor activity, immune response, and kinetic solubility : the impact of linker chemistry in conjugated NOD2/TLR4 agonists

Paradiso, Emiliano (author), Janež, Špela (author), Jakopin, Žiga (author)

Novel immunopotentiators are essential for advancing our understanding of immune receptor crosstalk and for addressing infectious diseases. Previous studies have suggested that coactivation of nucleotide-binding oligomerization domain-containing protein 2 (NOD2) and Toll-like receptor 4 (TLR4) can synergistically enhance the immune response. To investigate this synergy, we synthesized and evaluated a series of conjugated NOD2/TLR4 dual agonists comprising our in-house NOD2 agonist and two structurally distinct TLR4 agonists connected via flexible or rigid linkers. Our findings indicate that dual agonist activity toward both NOD2 and TLR4 is diminished upon conjugation. We also show that the linker chemistry significantly influences the kinetic solubility of these conjugates. Furthermore, the conjugates elicit distinct immunomodulatory effects in human primary peripheral blood mononuclear cells, characterized by a Th2-polarized cytokine response. These results provide insights into the structure–activity relationship of conjugated NOD2/TLR4 agonists and offer preliminary guidelines for tuning their solubility profiles.

• Language: English
• Keywords: agonists, conjugate acid-base pairs, ethyl groups, mixtures, solubility
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2025
• Number of pages: Str. 39060-39072
• Numbering: Vol. 10, iss. 34
• PID: 20.500.12556/RUL-176560
• UDC: 544.351.3
• ISSN on article: 2470-1343
• DOI: 10.1021/acsomega.5c05358
• COBISS.SI-ID: 253244675
• Publication date in RUL: 03.12.2025

Record is a part of a journal

• Title: ACS omega
• Shortened title: ACS omega
• Publisher: American Chemical Society
• ISSN: 2470-1343
• COBISS.SI-ID: 525873945

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: agonisti, konjugirani kislinsko-bazni pari, etilne skupine, zmesi, topnost, imunski odziv

Projects

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P1-0420
• Name: Napredna imunološka zdravila in celični pristopi v farmaciji

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: J3-4496
• Name: Adjuvansi naslednje generacije za mukozna cepiva