Pharmacokinetic and Pharmacodynamic Characterisation of an Anti-Mouse TNF Receptor 1 Domain Antibody Formatted for In Vivo Half-Life Extension.
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.
Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom.
Laura J
LJ
Goodall
Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom.
Milan
M
Ovecka
Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom.
Daniel
D
Rycroft
Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom.
Sarah L
SL
Friel
Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom.
Andrew
A
Sanderson
R&D Projects, Clinical Platforms and Sciences, GlaxoSmithKline, Stevenage, United Kingdom.
Prafull
P
Mistry
Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom.
Marie L
ML
Davies
Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom.
A Allart
AA
Stoop
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000276:immunology / Q000628:therapy*; D002460:Cell Line; D000939:Epitopes / Q000187:drug effects / Q000276:immunology; D006801:Humans; D015850:Interleukin-6 / Q000097:blood; D051379:Mice; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage*; D015398:Signal Transduction; D061905:Single-Domain Antibodies / Q000008:administration & dosage*; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; C000607805:DMS5540; D000939:Epitopes; D015850:Interleukin-6; D047888:Receptors, Tumor Necrosis Factor, Type I; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; C506236:Tnfrsf1a protein, mouse; D014409:Tumor Necrosis Factor-alpha