2. Neurokinin-1R (SP Receptor) Antagonists for HIV Therapy Steven D. Douglas, MD The Children’s Hospital of Philadelphia and University of Pennsylvania School of Medicine May 15, 2006 Supported by P01 MH-76388 and R01 MH-49981

3. CNS Substance P-Neurokinin-1 Receptor

4. Substance P ( Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH 2 )

5. Tachykinin Family TAC-1 (PPT-A) NK-1R: SP >NKA>NKB NK-2R: NKA>NKB>SP NK-3R: NKB>NKA>SP Substance PNeurokinin ANeurokinin BHemokinin-1 Endokinins TAC-3 (PPT-B)TAC-4 Page N.M. (2005) 26:1356

6. Differential binding sites for substance P and the nonpeptide antagonist CP-96,345 in the NK1 receptor. Hokfelt at al. Journal of Internal Medicine 249:27-40, 2001.

7. Neurokinin-1R antagonist(s) substance P – preferring potential therapeutic pathways 1. Anti-viral HIV – In vitro and in vivo Cellular mechanism 2. Immunomodulatory 3. Anti-depressive behavior “Neurokinin-1R (SP Receptor) Antagonists for HIV Therapy” 1 P01 MH76388

8. There is a bi-directional interaction between SP and HIV SP HIV Douglas, et al 2001. AIDS 15:2043-2045. Lai, et al 2001. Proc. Natl. Acad. Sci. USA. 98:3970-75. Ho, et al 2002. FASEB J. 16:616-618.

9. Effect of HIV (Bal) Infection on SP Production by Macrophages

10. Neurokinin-1 receptor (NK-1R) antagonists 1.Peptide and Non-peptide (e.g. Spantide) 2.NK-1R, dual, and pan-NK 3.Classification (groups): Diamines Amino-ethers Perhydroisoindoles Benzylpiperidine amides Quinoline and napthyridine amides Trytophan analogues

11. Lai et al. 2001, PNAS 98:3970-3975 NK-1R Antagonist (CP96,345) Inhibits HIV Infection of Macrophages The morphology of untreated and HIV-infected (A), CP-96,345- treated (10 -7 M) and infected (B), and untreated and uninfected MDM (C) was observed and photographed under a light microscopy (X 400) 12 days after infection.

12. Effect of CP-96,345 or RP-67,580 on CCR5 Expression in Monocytes  -actin CCR5 200 bp 500 bp -6 -8 CP-96,345 -6 -8 RP-67,580 Marker Control

13. Aprepitant Chemical Structure of Aprepitant Aprepitant: FDA approved drug for the treatment of chemotherapy induced nausea and vomiting (CINV)

14. SP receptor antagonist (Emend ® ) potently inhibits HIV (Bal) replication of human macrophages Wang X et al., poster presentation at the 13 th Conference on Retroviruses and Opportunistic Infectious (CROI), February 5-9, 2006, Denver, CO.

15. Inhibition of HIV (Bal) Infection of MDM by NK-1R Antagonists (10 -6 M)

16. Effect of Aprepitant on AZT Resistant Virus Infection of MDM A012 G691-6 and A018G901-6 were kind gifts from NIH AIDS Research and Reagent Program

17. Antiviral Effects of NK-1R Antagonists 1. Activity in MDM and PBMC 2. Down-regulation of CCR5 3. Activity against ART resistant strains

18. CCR5 NK-1R Signal Transduction? Hetero- dimerization? Protein Synthesis? CXCR4 HIV Cytokines, Chemokines, CCR5, CXCR4 NK-1R Antagonists NK-1R:CCR5 Interaction in Macrophages CD4 SP

19. SP enhanced CCL5-mediated calcium response via truncated NK-1R JP Lai et al. PNAS 130:7771-7776, 2006

20. Substance P antagonists down- regulate CCR5 expression CCR5  32 heterozygosity is associated with slower disease progression Lai, Jian-Ping et al. (2001) Proc. Natl. Acad. Sci. USA 98, 3970-3975 de Roda Husman, A.-M. et. al. Ann Intern Med 1997;127:882-890 CCR5 and SP/NK-1R

21. Program Project (P01 MH76388): Overall PI: SD Douglas CHOP, Douglas lab (Project 2) Steven D. Douglas, MD (PI) Jian-Ping Lai, MD Florin Tuluc, MD, PhD CHOP (Project 4) Jordan Orange, MD, PhD Donald E. Campbell, PhD Linda Shawver Nancy Tustin Nancy Raftery University of Pennsylvania (Project 4) Pablo Tebas, MD (PI) Dwight L. Evans, MD David F. Dinges, PhD David Gettes Tulane Nat’l Primate Research Center (Project 3) Andrew Lackner, PhD, DVM (PI) Kate Baker, PhD Pyone-Pyone Aye, PhD Lauren Cox, MA CHOP, Kilpatrick lab (Project 2) Laurie E. Kilpatrick, PhD Haiying Li Irene Chernova Seracare Bioservices, Inc.* (Core B) Janet L. Lathey, PhD (PI) Gabriela Tudor, PhD Lequan Nguyen *(private sector partner) CHOP, Pharmacology (Core C) Jeffrey S. Barrett, PhD (PI) Di Wu Mahesh Narayan CHOP, Biostatistics (Core C) Avital Cnaan, PhD (PI) Huaqing Zhao, PhD Michael Donaghue CHOP, Ho lab (Project 1) Wen-Zhe Ho, MD (PI) Xu Wang, MSc Li Song

23. Thank you! Douglas@email.chop.edu (215) 590-1978 (Office) (215) 590-3044 (Fax) Div. Of Allergy-Immunology Rm. 1208ARC, CHOP