A fully caninised anti-NGF monoclonal antibody for pain relief in dogs

Abstract

Background: Monoclonal antibodies are a major class of biological therapies in human medicine but have not yet been successfully applied to veterinary species. We have developed a novel approach, PETisation, to rapidly convert antibodies for use in veterinary species. As an example, anti-nerve growth factor (anti-NGF) monoclonal antibodies (mAbs) which are effective in reducing acute and chronic pain in rodents and man are potentially useful for treating pain in dogs but a fully caninised mAb is required in order to avoid an immune response. The aim of this study was to determine the optimal properties of a caninised anti-NGF mAb for safe, repeated administration to dogs, to determine its pharmacokinetic properties and to evaluate its efficacy in a model of inflammatory pain in vivo. Results: Starting with a rat anti-NGF mAb, we used a novel algorithm based on expressed canine immunoglobulin sequences to design and characterise recombinant caninised anti-NGF mAbs. Construction with only 2 of the 4 canine IgG heavy chain isotypes (A and D) resulted in stable antibodies which bound and inhibited NGF with high-affinity and potency but did not bind complement C1q or the high-affinity Fc receptor gamma R1 (CD64). One of the mAbs (NV-01) was selected for scale-up manufacture, purification and pre-clinical evaluation. When administered to dogs, NV-01 was well tolerated, had a long serum half-life of 9 days, was not overtly immunogenic following repeated dosing in the dog and reduced signs of lameness in a kaolin model of inflammatory pain.

Identification of a candidate therapeutic antibody for treatment of canine B-cell lymphoma

“…To our knowledge, this is the first report comparing the in vivo behavior of different canine isotypes. While the exact mechanism(s) of B-cell depletion by the 1E4-cIgGB and 1E4-cIgGC anti-CD20 antibodies we have developed in this study are under investigation, our B-cell depletion and CD16a binding results are consistent with the findings of recent reports of in vitro characterization of the four canine isotypes; these studies also found that only cIgGB and cIgGC are able to engage receptors involved in ADCC and CDC (Bergeron et al, 2014;Gearing et al, 2013) Together, these findings should pave the way for custom design and engineering of canine biologics.…”

In Vitro and In Vivo Characterization of a Fully Felinized Therapeutic Anti‐Nerve Growth Factor Monoclonal Antibody for the Treatment of Pain in Cats

“…To decrease the immunogenic potential of rat anti‐mouse NGF mAb αD11 in the cat, while retaining its high affinity for NGF, amino acid substitutions were made to the heavy and light chain variable domain framework sequences by alignment with a collection of predicted protein sequences encoded by expressed feline immunoglobulin (IgG) complementary deoxyribonucleic acid (cDNA) sequences. This approach (that we refer to as PETization) previously was used to generate a fully caninized anti‐NGF mAb that has a promising efficacy and safety profile in dogs . Where an amino acid in the αD11 sequence corresponded to an amino acid in the collection no change was made.…”

“…Plates were coated with 2.5 μg/mL mouse NGF and blocked with 5% bovine serum albumin (BSA)/phosphate‐buffered saline (PBS). Coated wells were incubated for 1 hour at room temperature with NV‐02 antibody, or as a positive control a caninized anti‐NGF mAb with a IgG‐B isotype (complement‐binding) heavy chain, diluted in PBS/1% BSA. Antibody concentrations ranged from 10 μg/mL to 1.0 μg/mL.…”

“…A Langmuir curve fit model then was used to determine the specific affinity. Inhibition of NGF binding by NV‐02 was assessed using TF‐1 cells as previously described . TF‐1 cells were starved for 24 hours, then cultured in 96‐well plates in media supplemented with 1 ng/mL mouse NGF and increasing concentrations of NV‐02 or caninized anti‐NGF mAb NV‐01.…”

“…Anti‐nerve growth factor (NGF) monoclonal antibodies (mAbs) have been shown to have analgesic effects in rodent models of pain, in several human clinical trials and, more recently, in proof‐of‐concept clinical studies in osteoarthritic dogs . Long‐acting pain relief (>4 weeks) and good tolerability was observed in the dog studies after a single injection.…”