1. “A Fully Caninised Anti-NGF Monoclonal Antibody for Pain Relief in Dogs” Scientific Paper Presentation Madeline Jackson 17418057 Gearing, D. P., Virtue, E. R., Gearing, R. P., & Drew, A. C. (2013). A fully caninised anti-NGF monoclonal antibody for pain relief in dogs. BMC veterinary research, 9(1), 226.

2. Introduction Monoclonal antibodies (mAb) Anti-NGF mAbs reduce pain in rodents and in humans Potential to reduce pain in dogs New technique called ‘PETisation’ – Fully caninised anti-NGF mAb (NV-01)

3. Introduction Nerve Growth Factor (NGF) – Promotes pain and hyperalgesia – Produced by a variety of inflammatory and immune cells – Binds to trkA receptor on nociceptors – Causes sprouting of nerve endings into inflammation site – Mutations in NGF can reduce pain responses Anti-NGF mAb acts as an analgesic in rodents and humans – Arthritis, cancer, and bone fracture pain

4. Aim “To determine the optimal properties of a caninised anti-NGF mAb for safe, repeated administration to dogs; to determine its pharmacokinetic properties; and the evaluate its efficacy in a model of inflammatory pain in vivo”

5. Methods Used purified mouse NGF Conversion of αD11 variable domain of IgG – PETisation NV-01 produced by recombinant Ab preparations Anti-NGF mAb detection by ELISA Kaolin model of inflammatory pain – Group 1 – negative control – Group 2 – NV-01 IV – Group 3 – NV-01 SC – Group 4 – positive control – meloxicam (given daily)

6. Results PETisation – was successful in fully caninising anti-NGF mAbs Four isotypes tested – A and D – no activation of complement pathway – B and C – activation of complement pathway – NOT USED – A and D – no binding to canine CD64 – B and C – binding to canine CD64– NOT USED No adverse effects observed

7. Results Group 1 – remained lame Group 2 – matched results of meloxicam, but took longer Group 3 – delayed reduction in lameness Group 4 – reduced average lameness Group 1 – Group 2 – Group 3 – Group 4 –

8. Discussion PETisation – Simpler and more effective way method of the conversion of rodent Ab to canine Ab Fully caninised mAb – αD11 domain is well described and has a high affinity for mouse and human NGF – Safe to use – isotypes A and D NV-01 is just as effective as meloxicam – Safe, with no observable side effects – 9 day half life and can be administered repeatedly without inducing a neutralising immune response

9. Bigger Picture Several experiments in the past few years have found similar results to this one – Ralph et al. (2014) concluded that an anti-NGF mAb decreases pain caused by osteoarthritis in dogs – Tiseo et al. (2014) conducted a study on the effect of an anti-NGF mAb on the reduction of osteoarthritis pain in humans and found that it was an effective treatment – Ishikawa et al. (2015) also found that using an anti-NGF mAb can provide a long-lasting analgesic effect in a rat model with osteoarthritis – Lascelles et al. (2015) performed a study that was an extension of this analysed experiment and concluded that chronic pain in dogs could be reduced with an anti-NGF mAb

10. Conclusion NV-01 has been deemed safe for repeated use in dogs, with no observable side effects, a long half life, and for effective pain relief. These preliminary studies suggests that this novel fully caninised anti-NGF mAb shows considerable potential as an analgesic for dogs.

11. References Ishikawa, G., Koya, Y., Tanaka, H., & Nagakura, Y. (2015). Long-term analgesic effect of a single dose of anti-NGF antibody on pain during motion without notable suppression of joint edema and lesion in a rat model of osteoarthritis. Osteoarthritis and Cartilage. Lascelles, B. D., Knazovicky, D., Case, B., Freire, M., Innes, J. F., Drew, A. C., & Gearing, D. P. (2015). A canine-specific anti-nerve growth factor antibody alleviates pain and improves mobility and function in dogs with degenerative joint disease- associated pain. BMC veterinary research, 11(1), 101. Tiseo, P. J., Kivitz, A. J., Ervin, J. E., Ren, H., & Mellis, S. J. (2014). Fasinumab (REGN475), an antibody against nerve growth factor for the treatment of pain: results from a double-blind, placebo-controlled exploratory study in osteoarthritis of the knee. PAIN®, 155(7), 1245-1252. Webster, R. P., Anderson, G. I., & Gearing, D. P. (2014). Canine brief pain inventory scores for dogs with osteoarthritis before and after administration of a monoclonal antibody against nerve growth factor. American journal of veterinary research, 75(6), 532-535.