Pharmacokinetic and Pharmacodynamic Characterisation of an Anti-Mouse TNF Receptor 1 Domain Antibody Formatted for In Vivo Half-Life Extension

• Published in: PloS one 2015-09, Vol.10 (9), p.e0137065-e0137065
• Main Authors: Goodall, Laura J, Ovecka, Milan, Rycroft, Daniel, Friel, Sarah L, Sanderson, Andrew, Mistry, Prafull, Davies, Marie L, Stoop, A Allart
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Antibodies, Monoclonal - administration & dosage; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal - pharmacokinetics; Apoptosis; Arthritis; Arthritis, Rheumatoid - blood; Arthritis, Rheumatoid - immunology; Arthritis, Rheumatoid - therapy; Binding; Biocompatibility; Cell Line; Chromatography; Chronic illnesses; Cytotoxicity; Drug delivery systems; Epitopes; Epitopes - drug effects; Epitopes - immunology; Genetic aspects; Half-life; Homeostasis; Humans; Immune clearance; Immunoglobulins; In vivo methods and tests; Inhibition; Interleukin 6; Interleukin-6 - blood; Life extension; Ligands; Medical treatment; Mice; Monoclonal antibodies; Pharmacodynamics; Pharmacokinetics; Pharmacology; Physiological aspects; Receptors; Receptors, Tumor Necrosis Factor, Type I - antagonists & inhibitors; Receptors, Tumor Necrosis Factor, Type I - genetics; Receptors, Tumor Necrosis Factor, Type I - immunology; Recombinant Fusion Proteins - administration & dosage; Rheumatoid arthritis; Signal Transduction; Single-Domain Antibodies - administration & dosage; Toxicity; Tumor necrosis factor; Tumor necrosis factor receptors; Tumor Necrosis Factor-alpha - antagonists & inhibitors; Tumor Necrosis Factor-alpha - immunology; Tumor Necrosis Factor-alpha - metabolism; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Tumors
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0137065

Tumour Necrosis Factor-α (TNF-α) inhibition has been transformational in the treatment of patients with inflammatory disease, e.g. rheumatoid arthritis. Intriguingly, TNF-α signals through two receptors, TNFR1 and TNFR2, which have been associated with detrimental inflammatory and beneficial immune-regulatory processes, respectively. To investigate if selective TNFR1 inhibition might provide benefits over pan TNF-α inhibition, tools to investigate the potential impact of pharmacological intervention are needed. Receptor-deficient mice have been very insightful, but are not reversible and could distort receptor cross-talk, while inhibitory anti-TNFR1 monoclonal antibodies have a propensity to induce receptor agonism. Therefore, we set out to characterise a monovalent anti-TNFR1 domain antibody (dAb) formatted for in vivo use. The mouse TNFR1 antagonist (DMS5540) is a genetic fusion product of an anti-TNFR1 dAb with an albumin-binding dAb (AlbudAb). It bound mouse TNFR1, but not human TNFR1, and was an antagonist of TNF-α-mediated cytotoxicity in a L929 cell assay. Surprisingly, the dAb did not compete with TNF-α for TNFR1-binding. This was supported by additional data showing the anti-TNFR1 epitope mapped to a single residue in the first domain of TNFR1. Pharmacokinetic studies of DMS5540 in mice over three doses (0.1, 1.0 and 10 mg/kg) confirmed extended in vivo half-life, mediated by the AlbudAb, and demonstrated non-linear clearance of DMS5540. Target engagement was further confirmed by dose-dependent increases in total soluble TNFR1 levels. Functional in vivo activity was demonstrated in a mouse challenge study, where DMS5540 provided dose-dependent inhibition of serum IL-6 increases in response to bolus mouse TNF-α injections. Hence, DMS5540 is a potent mouse TNFR1 antagonist with in vivo pharmacokinetic and pharmacodynamic properties compatible with use in pre-clinical disease models and could provide a useful tool to dissect the individual contributions of TNFR1 and TNFR2 in homeostasis and disease.