Fexofenadine inhibits TNF signaling through targeting to cytosolic phospholipase A2 and is therapeutic against inflammatory arthritis

Abstract

ObjectiveTumour necrosis factor alpha (TNF-α) signalling plays a central role in the pathogenesis of various autoimmune diseases, particularly inflammatory arthritis. This study aimed to repurpose clinically approved drugs as potential inhibitors of TNF-α signalling in treatment of inflammatory arthritis.MethodsIn vitro and in vivo screening of an Food and Drug Administration (FDA)-approved drug library; in vitro and in vivo assays for examining the blockade of TNF actions by fexofenadine: assays for defining the anti-inflammatory activity of fexofenadine using TNF-α transgenic (TNF-tg) mice and collagen-induced arthritis in DBA/1 mice. Identification and characterisation of the binding of fexofenadine to cytosolic phospholipase A2 (cPLA2) using drug affinity responsive target stability assay, proteomics, cellular thermal shift assay, information field dynamics and molecular dynamics; various assays for examining fexofenadine inhibition of cPLA2 as well as the dependence of fexofenadine’s anti-TNF activity on cPLA2.ResultsSerial screenings of a library composed of FDA-approved drugs led to the identification of fexofenadine as an inhibitor of TNF-α signalling. Fexofenadine potently inhibited TNF/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-ĸB) signalling in vitro and in vivo, and ameliorated disease symptoms in inflammatory arthritis models. cPLA2 was isolated as a novel target of fexofenadine. Fexofenadine blocked TNF-stimulated cPLA2 activity and arachidonic acid production through binding to catalytic domain 2 of cPLA2 and inhibition of its phosphorylation on Ser-505. Further, deletion of cPLA2 abolished fexofenadine’s anti-TNF activity.ConclusionCollectively, these findings not only provide new insights into the understanding of fexofenadine action and underlying mechanisms but also provide new therapeutic interventions for various TNF-α and cPLA2-associated pathologies and conditions, particularly inflammatory rheumatic diseases.

Comparative Analysis of Compatibility Influence on Invigorating Blood Circulation for Combined Use of Panax Notoginseng Saponins and Aspirin Using Metabolomics Approach

“…Arachidonic acid is released from the membrane via the cleavage by phospholipase A2 in response to stimuli (Burstein et al, 1982;Liu et al, 2019). Thromboxane A2, a product of arachidonic acid with the aid of cyclooxygenase, is involved in vascular contraction and has been implicated in platelet activation (Rosenfeld et al, 2010).…”

Dynamic protein quantitation (DyProQ) of procollagen-I by CRISPR-Cas9 NanoLuciferase tagging

“…A proposed mechanism of action is reduced TGF-β stimulated collagen secretion by primary lung fibroblasts 34 . Others included Thioridazine, a first-generation antipsychotic drug and a potential anti-inflammatory 36 ; Mefloquinine, used to prevent malaria but has suspected psychotic side effects 37 ; CEP-18770 (Delanzomib), a proteasome inhibitor with potential anti-tumour activity 38 ; Dacomitinib, an EGFR tyrosine kinase inhibitor being investigated for treatment of non-small-cell lung cancer; LDK378 (Ceritinib), a potent ALK inhibitor used in the treatment of non-small-cell lung cancer 39 ; Terfenadine (which produces the metabolite, Fexofenadine) is a second-generation antihistamine that inhibits TNF signalling and is a therapeutic against inflammatory arthritis 40 ; Fingolimod is a first-in-class sphingosine-1-phosphate receptor modulator and immunosuppressant that is approved for the treatment of relapsing-remitting multiple sclerosis 41 ; AZD-9291 (Osimertinib), is a mutant-selective EGFR inhibitor used to treat non-small-cell lung cancer 42 and, Benserazide is a peripheral decarboxylase inhibitor that increases the amount of levodopa crossing into the brain and its subsequent conversion to dopamine, and is used in combination with other drugs in the treatment of Parkinson’s disease. We propose that these 10 compounds are candidates for repurposing in the treatment of fibrosis.…”

A20: a master regulator of arthritis

“…As mentioned above, TNF-α inhibitors have been frequently used to treat severe RA [ 38 ] because of the pioneering status of TNF-α at the apex of the pro-inflammatory cytokine cascade [ 6 , 41 ]. Moreover, the activation of TNF-induced TNFR signaling and downstream signaling evoked by TNFR, such as NF-κB and MAPK, implicates a variety of responses.…”