Transcript Slide 1

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
CNS
Substance P-Neurokinin-1 Receptor
Substance P
(Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2)
Tachykinin Family
TAC-1 (PPT-A)
Substance P
Neurokinin A
TAC-3 (PPT-B)
TAC-4
Neurokinin B
Hemokinin-1
Endokinins
NK-1R: SP>NKA>NKB
NK-2R: NKA>NKB>SP
NK-3R: NKB>NKA>SP
Page N.M. (2005) 26:1356
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.
Neurokinin-1R antagonist(s) substance P –
preferring potential therapeutic pathways
“Neurokinin-1R (SP Receptor)
Antagonists for HIV Therapy”
1 P01 MH76388
1. Anti-viral HIV – In vitro and in vivo
Cellular mechanism
2. Immunomodulatory
3. Anti-depressive behavior
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.
Effect of HIV (Bal) Infection on SP Production
by Macrophages
45
5000
40
RT
35
4000
3000
30
2000
25
20
1000
15
0
10
5
2.5
1
HIV Bal Inoculum (p24:ng)
0
RT Activity (CPM/5ul)
Substance P (pg/ml)
SP
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
NK-1R Antagonist (CP96,345) Inhibits HIV Infection of
Macrophages
A
B
C
Fig. 5
The morphology of untreated and HIV-infected (A), CP-96,345treated (10-7M) and infected (B), and untreated and uninfected
MDM (C) was observed and photographed under a light
microscopy (X 400) 12 days after infection.
Lai et al. 2001, PNAS 98:3970-3975
Marker
Effect of CP-96,345 or RP-67,580 on CCR5
Expression in Monocytes
200
bp
500 bp
CP-96,345 RP-67,580
-6
-8
-6
-8
CCR5
b-actin
Chemical Structure of Aprepitant
Aprepitant
Aprepitant: FDA approved drug for the treatment of chemotherapy
induced nausea and vomiting (CINV)
SP receptor antagonist (Emend®) potently inhibits
HIV (Bal) replication of human macrophages
70000
Control
E 1nM
E 100 nM
E 10 uM
HIV RT activity
60000
50000
40000
30000
20000
10000
0
4
8
12
16
20
Days post-infection
Wang X et al., poster presentation at the 13th Conference on Retroviruses and
Opportunistic Infectious (CROI), February 5-9, 2006, Denver, CO.
Inhibition of HIV (Bal) Infection of MDM
by NK-1R Antagonists (10-6 M)
25
20
15
10
5
33
06
0
L7
67
,5
80
R
P
96
,3
45
PC
J1
2,
25
5
C
pr
ep
i ta
nt
A
on
tr
ol
0
C
HIV RT Activity (103 cpm)
30
Effect of Aprepitant on AZT Resistant Virus
Infection of MDM
AZT Resistant HIV-1 (A012 G691-6)
AZT Resistant HIV-1 (A018 G901-6)
300
20
HIV P24 Level (pg/ml)
Aprepitant
Control
3
HIV P24 Level (x10 pg/ml)
24
16
12
8
4
Aprepitant
250
Control
200
150
100
50
0
0
Day 0
Day 3
Day 5
Day7
Day 0
Day 3
Day 5
Day7
A012 G691-6 and A018G901-6 were kind gifts from NIH AIDS
Research and Reagent Program
Antiviral Effects of NK-1R
Antagonists
1. Activity in MDM and PBMC
2. Down-regulation of CCR5
3. Activity against ART resistant
strains
NK-1R:CCR5 Interaction in Macrophages
SP
Heterodimerization?
HIV
CCR5
NK-1R
Antagonists
NK-1R
CD4
Signal Transduction?
CXCR4
Protein Synthesis?
Cytokines, Chemokines,
CCR5, CXCR4
SP enhanced CCL5-mediated calcium
response via truncated NK-1R
250
Undifferentiated
THP-1
[Ca 2+]i nM
200
CCL5
Aprepitant + SP + CCL5
SP + CCL5
150
-8
CCL5 (10 M)
-6
SP (10 M)
100
50
0
0
50
100
150
200
Time (seconds)
JP Lai et al. PNAS 130:7771-7776, 2006
250
300
CCR5 and SP/NK-1R
• Substance P
antagonists downregulate CCR5
expression
Lai, Jian-Ping et al. (2001) Proc. Natl. Acad. Sci. USA 98,
3970-3975
• CCR5 D32
heterozygosity is
associated with slower
disease progression
de Roda Husman, A.-M. et. al. Ann Intern Med 1997;127:882-890
Program Project (P01 MH76388):
Overall PI: SD Douglas
CHOP, Ho lab (Project 1)
Wen-Zhe Ho, MD (PI)
Xu Wang, MSc
Li Song
CHOP, Douglas lab (Project 2)
Steven D. Douglas, MD (PI)
Jian-Ping Lai, MD
Florin Tuluc, MD, PhD
CHOP, Kilpatrick lab
(Project 2)
Laurie E. Kilpatrick, PhD
Haiying Li
Irene Chernova
Tulane Nat’l Primate Research
Center (Project 3)
Andrew Lackner, PhD, DVM (PI)
Kate Baker, PhD
Pyone-Pyone Aye, PhD
Lauren Cox, MA
University of Pennsylvania
(Project 4)
Pablo Tebas, MD (PI)
Dwight L. Evans, MD
David F. Dinges, PhD
David Gettes
CHOP (Project 4)
Jordan Orange, MD, PhD
Donald E. Campbell, PhD
Linda Shawver
Nancy Tustin
Nancy Raftery
Seracare Bioservices, Inc.*
(Core B)
Janet L. Lathey, PhD (PI)
Gabriela Tudor, PhD
Lequan Nguyen
CHOP, Biostatistics (Core C)
Avital Cnaan, PhD (PI)
Huaqing Zhao, PhD
Michael Donaghue
CHOP, Pharmacology
(Core C)
Jeffrey S. Barrett, PhD (PI)
Di Wu
Mahesh Narayan
*(private sector partner)
Thank you!
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