tetano
Editor, Senior Moderator
Innate Immun
. 2025 Jan-Dec:31:17534259241313201.
doi: 10.1177/17534259241313201. Preclinical development of the TLR4 antagonist FP12 as a drug lead targeting the HMGB1/MD-2/TLR4 axis in lethal influenza infection
Kari Ann Shirey[SUP] 1 [/SUP], Alessio Romerio[SUP] 2 [/SUP], Mohammed Monsoor Shaik[SUP] 2 [/SUP], David S Leake[SUP] 3 [/SUP], Charys Palmer[SUP] 4 [/SUP], Natalia Skupinska[SUP] 4 [/SUP], Jules Paton[SUP] 1 [/SUP], Grisha Pirianov[SUP] 4 [/SUP], Jorge Cg Blanco[SUP] 5 [/SUP], Stefanie N Vogel[SUP] 1 [/SUP], Francesco Peri[SUP] 2 [/SUP]
Affiliations
Background: Acute Lung Injuries (ALI) are a severe consequence of influenza-induced cytokine storm that can cause respiratory failure and death. It has been demonstrated that Toll-like Receptor 4 (TLR4) is involved in cytokine storm and that TLR4[SUP]-/-[/SUP] mice are protected against ALI. Therefore, TLR4 is a prime target for protection against ALI. FP12 is a known TLR4 antagonist that reduces TLR4-dependent immune activation and it is a promising lead compound for the treatment of innate immunity related pathologies.
Objectives: We present here the preclinical development of FP12 as an anti-inflammatory lead compound acting on influenza-induced ALI.
Methods: In vitro: We pre-treated THP-1 cells with FP12 (10 μM) for 0.5 h, then exposed to LPS (100 ng/ml) for 0 to 16 h. In some experiments, cells were simultaneously incubated with FP12 and LPS, or FP12 was added 30 min after LPS. Cytokine levels were measured by Western blot and ELISA assays. In vivo: WT C57BL/6J mice were infected with mouse-adapted influenza virus (PR8). Two days after infection, mice received either vehicle, FP7 (200 µg/mouse), or FP12 (200 µg/mouse) once daily (Day 2 to Day 6). Mice were monitored daily for survival for 14 days. Data were collected through histological staining, qRT-PCR, and ELISA assay.
Results: FP12 treatment inhibited both LPS- and HMGB1-induced TLR4 intracellular pathways (MyD88 and TRIF) leading to significantly reduced levels of a variety of proinflammatory cytokines including Type I interferon (IFN-β), highlighting its effectiveness in controlling proinflammatory protein production and reducing inflammation. FP12 protected mice therapeutically from influenza virus-induced lethality and reduced both cytokine gene expression and High Mobility Group Box 1 (HMGB1) levels in the lungs as well as ALI.
Conclusion: FP12 can antagonize TLR4 activation in vitro and protects mice from severe influenza infection, most likely by reducing the TLR4-dependent cytokine storm mediated by danger-associated molecular patterns (DAMPs).
Keywords: ALI; Drug discovery; HMGB1; TLR4; influenza.
. 2025 Jan-Dec:31:17534259241313201.
doi: 10.1177/17534259241313201. Preclinical development of the TLR4 antagonist FP12 as a drug lead targeting the HMGB1/MD-2/TLR4 axis in lethal influenza infection
Kari Ann Shirey[SUP] 1 [/SUP], Alessio Romerio[SUP] 2 [/SUP], Mohammed Monsoor Shaik[SUP] 2 [/SUP], David S Leake[SUP] 3 [/SUP], Charys Palmer[SUP] 4 [/SUP], Natalia Skupinska[SUP] 4 [/SUP], Jules Paton[SUP] 1 [/SUP], Grisha Pirianov[SUP] 4 [/SUP], Jorge Cg Blanco[SUP] 5 [/SUP], Stefanie N Vogel[SUP] 1 [/SUP], Francesco Peri[SUP] 2 [/SUP]
Affiliations
- PMID: 40033742
- DOI: 10.1177/17534259241313201
Background: Acute Lung Injuries (ALI) are a severe consequence of influenza-induced cytokine storm that can cause respiratory failure and death. It has been demonstrated that Toll-like Receptor 4 (TLR4) is involved in cytokine storm and that TLR4[SUP]-/-[/SUP] mice are protected against ALI. Therefore, TLR4 is a prime target for protection against ALI. FP12 is a known TLR4 antagonist that reduces TLR4-dependent immune activation and it is a promising lead compound for the treatment of innate immunity related pathologies.
Objectives: We present here the preclinical development of FP12 as an anti-inflammatory lead compound acting on influenza-induced ALI.
Methods: In vitro: We pre-treated THP-1 cells with FP12 (10 μM) for 0.5 h, then exposed to LPS (100 ng/ml) for 0 to 16 h. In some experiments, cells were simultaneously incubated with FP12 and LPS, or FP12 was added 30 min after LPS. Cytokine levels were measured by Western blot and ELISA assays. In vivo: WT C57BL/6J mice were infected with mouse-adapted influenza virus (PR8). Two days after infection, mice received either vehicle, FP7 (200 µg/mouse), or FP12 (200 µg/mouse) once daily (Day 2 to Day 6). Mice were monitored daily for survival for 14 days. Data were collected through histological staining, qRT-PCR, and ELISA assay.
Results: FP12 treatment inhibited both LPS- and HMGB1-induced TLR4 intracellular pathways (MyD88 and TRIF) leading to significantly reduced levels of a variety of proinflammatory cytokines including Type I interferon (IFN-β), highlighting its effectiveness in controlling proinflammatory protein production and reducing inflammation. FP12 protected mice therapeutically from influenza virus-induced lethality and reduced both cytokine gene expression and High Mobility Group Box 1 (HMGB1) levels in the lungs as well as ALI.
Conclusion: FP12 can antagonize TLR4 activation in vitro and protects mice from severe influenza infection, most likely by reducing the TLR4-dependent cytokine storm mediated by danger-associated molecular patterns (DAMPs).
Keywords: ALI; Drug discovery; HMGB1; TLR4; influenza.