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

Abstract

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.

Apelin peptides linked to anti‐serum albumin domain antibodies retain affinity in vitro and are efficacious receptor agonists in vivo

“…Furthermore, we have conjugated an apelin mimetic peptide to an anti‐serum albumin domain antibody (AlbudAb) to extend transit time in the blood. AlbudAbs have previously been conjugated to soluble IL‐1Rα, GLP‐1 and TNF‐R1 and demonstrated half‐life extension with retained activity. To our knowledge, this is the first time the AlbudAb moiety has been chemically rather than genetically conjugated to a ligand.…”

Antibody engineering & therapeutics, the annual meeting of the antibody society December 7–10, 2015, San Diego, CA, USA

“…Pharmacokinetic studies of DMS5540 in mice over 3 doses (0.1, 1.0 and 10 mg/kg) confirmed extended in vivo half-life, mediated by the AlbudAb. 34 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 injection.…”

“…DMS5540 corresponds to a mouse TNFR1 antagonist, constituted by the genetic fusion product of an anti-TNFR1 dAb with an albumin-binding dAb (AlbudAb). 34 It bound mouse TNFR1, but not human TNFR1, and was an antagonist of TNF-mediated cytotoxicity in a L929 cell assay. 34 Surprisingly, the dAb did not compete with TNF for TNFR1-binding.…”

“… 34 It bound mouse TNFR1, but not human TNFR1, and was an antagonist of TNF-mediated cytotoxicity in a L929 cell assay. 34 Surprisingly, the dAb did not compete with TNF for TNFR1-binding. Pharmacokinetic studies of DMS5540 in mice over 3 doses (0.1, 1.0 and 10 mg/kg) confirmed extended in vivo half-life, mediated by the AlbudAb.…”

Injectables and Depots to Prolong Drug Action of Proteins and Peptides

“…An IL-1 receptor antagonist fused to AlbudAb showed a 130-fold longer half-life in vivo compared to the unmodified antagonist when tested in a mouse model of arthritis [ 194 ]. Other applications of the AlbudAb technology have also been reported for IFN-α2 [ 195 ] and a mouse anti-TNF receptor 1 antagonist [ 196 ]. An AlbudAb fusion to exendin-4 has been tested in phase I human trials by GlaxoSmithKline (GSK), who now own the AlbudAb technology [ 197 , 198 ].…”