1. Prostatic acid phosphatase (PAP) - a potent analgesic for treating pain Mark J. Zylka University of North Carolina at Chapel Hill

2. More Americans suffer from chronic pain than heart disease, diabetes and cancer combined Neuropathic pain Neuropathic pain nerve injury, shingles, diabetic neuropathy (~15 million Americans) Inflammatory and surgical pain Inflammatory and surgical pain (~27 million Americans have arthritis of knee)

3. Prostatic Acid Phosphatase (PAP) 1. Found in pain-sensing neurons 1. Found in pain-sensing neurons (makes it a “natural” product) Pain-sensing neuronsSpinal Cord 2. Drug-like 2. Drug-like (Use secreted version as drug; like insulin) 3. Recombinant PAP 3. Recombinant PAP (can produce in yeast, secreted into medium) * Provides clear path to FDA-approval 4. Potently suppresses pain 4. Potently suppresses pain (>8x better than morphine)

4. Possible Markets 1.Orthopedic surgery (prophylactic; preemptive analgesia) 2.Neuropathic pain, including postherpetic neuralgia 3.Lower back pain (inflammatory pain) 4.Cancer pain

5. Preclinical Data in Mice

6. Thermal Thermal test

7. Mechanical Test Mechanical

8. Analgesic effects last 3 days

9. PAP is >8x more effective than morphine; no side effects

10. PAP suppresses chronic inflammatory pain ThermalMechanical ….as well as Neuropathic pain (not shown)

11. (No desensitization) Repeated injections are effective (No desensitization) Makes long-term treatment of pain possible (multiple injections or intrathecal pump = repeated sales) Gene Therapy (PAP in Adenoassociated virus; rAAV) Non-injured paw is not affected. (suggests PAP selectively relieves pain in body part that hurts)

12. PAP as prophylactic – inject prior to surgery to prevent post-op pain ThermalMechanical

13. PAP works by making adenosine AMP (Adenosine monophosphate) ADO + P (Adenosine) PAP (Adenosine receptor knockout mice)

14. Adenosine is analgesic in humans

15. Fukunaga et al (2003) Pain 101:129 Preemptive analgesia Preemptive analgesia: i.v. adenosine during surgery provides long lasting pain relief in humans (much better than an opioid)

16. In collaboration w/ Stephen Frye’s group at UNC Inactive Prodrug-P (UNC543) Active drug + P PAP PAP-selective prodrugs (orally active)

17. Use of PAP as an analgesic 1.Use as biologic a. Acute: Intrathecal injection b. Chronic: repeated injections or in intrathecal pump (like morphine, Prialt) 2.PAP-selective prodrugs (orally active)

18. U. Helsinki (Finland) Pirkko Vihko Annakaisa Herrala Acknowledgments Support UNC Startup Funds, Sloan Foundation, Whitehall, NARSAD, Klingenstein, Searle, Rita Allen, NINDS Nate SowaBonnie Taylor-Blake Jennifer Coleman Yvette Chuang University of North Carolina Stephen Frye Jian Jin Julie Hurt

19. PAP purification from yeast (Pichia)