Overarching Goals

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Substance P Receptor (Neurokinin-1R)
Antagonists as Therapeutic Agents in HIV
Steven D. Douglas, MD August 4, 2004
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Overall Goal
To demonstrate the mechanism and activity of
neurokinin-1R (substance P) antagonists as
antiviral agents in HIV.
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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
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Projects and Cores

• Projects
• Anti-HIV Mechanisms of Neurokinin-1 Receptor
• (NK-1R) Antagonists Ho
• 2. Cellular Mechanisms and Interaction of
  Neurokinin-1
• Receptor (NK-1R) Antagonists and HIV Co-Receptors
  
• CCR5 and CXCR4 Douglas, Kilpatrick
• 3. SIV models of neurobehavioral, antiviral, and
• immunomodulatory SP antagonist(s) effects
  Lackner,
• Baker
• 4. Human Studies of Neurokinin-1 Receptor (NK-1R)
  
• Antagonists in HIV-1 - Tebas

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Projects and Cores

• Cores
• Administrative Douglas
• B. HIV Antiretroviral Drug Susceptibility and
• Drug Interactions Lathey
• C. Biostatistics and Pharmacology - Cnaan

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IPCP Organizational Schema
Core A Administration S. Douglas, Dir
Internal Advisory Board
External Advisory Board
Basic
Pre-clinical/clinical
P1 W-Z Ho, PI
P3 A. Lackner, PI
P2 S. Douglas, PI
P4 P. Tebas, PI
Core B Virus Susceptibility J Lathey, Dir BBI
(Private Sector Partner)
Core C Biostatistics and Pharmacology A. Cnaan,
Dir.
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Private Sector Partner
Name Boston Biomedica, Inc. Gaithersburg,
MD PI Dr. Janet Lathey HIV Antiretroviral Drug
Susceptibility and Drug Interactions Core
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Biotech Overview

• The research and development arm of the Company
  for Molecular Biology, Virology and Immunology
• Experienced Scientific Staff consisting of 9
  PhDs and more than 50 scientists
• Provides a variety of products and services to
  BBI operating units and other outside customers
• Specialty reagents and molecular and cellular
  biology services
• Blood and tissue processing and repository
  services
• Clinical trials for domestic and foreign test kit
  and device manufacturers
• Services typically provided under multi-year
  contracts
• Services focused in advanced biomedical research
  areas

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Virology Services Available for Clinical Trials
of Therapeutics

• HIV Antigen Detection and Quantitation
• HIV Culture and Viral Isolation
• Titration of Infectious Virus
• Drug Susceptibility Assays
• Viral Nucleic Acid Isolation
• Viral Nucleic Acid Quantitation

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Collaborative Resources

• CFAR Cores Douglas, Tebas, Lackner (Hoxie,
  Gonzalez)
• AACTU Tebas (PI)
• HUP GCRC Tebas
• HPTN Douglas, Metzger
• PPACTU Douglas (PI)
• CHOP Cores Molecular, Flow Cytometry,
• Biostatistics, Protein, Nucleic Acid

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Background and Prior NIMH-supported Work by
Investigators
MH49981
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CNS
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Substance P (Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-
Leu-Met-NH2)

• SP, described by von Euler and Gaddum in 1931,
  was the first neuropeptide to be identified.
• SP is a neuropeptide comprised of 11 amino acid
  with a wide-spread distribution in the central
  and peripheral nervous systems (Chang and Leeman,
  1970, 1971).
• SP belongs to the tachykinin family that includes
  SP, neurokinin A (SK), and neurokinin B.

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Biological Activities of SP

• A potent neurotransmitter and neuromodulator.
  Interactions with other neurotransmitters in the
  brain.
• Transmission of painful stimuli from periphery.
• Control blood flow (vasodilatation).
• Gastrointestinal motility, enteric secretion and
  absorption.
• A potent mediator of neuroimmunoregulation.
  Activation of cells of the immune system and
  participation of inflammatory process.
• A hematopoietic modulator.

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Tachykinin Family
Tachykinin
PPT-A
PPT-B
Substance P Neurokinin A
Neurokinin B
NK-1R SPgtNKAgtNKB NK-2R NKAgtNKBgtSP NK-3R
NKBgtNKAgtSP
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Human Immune Cells Express Substance P and Its
Receptor

• Ho, et al 1997. J. Immunol. 1595654-60.
• Lai, et al 1998. J. Neuroimmunol. 8680-6.
• Lai, et al 2000. Neuroscience 1011137-44.
• Li, et al 2000. J. Hematotherapy Stem Cell Res.
  9445.
• Lai, et al 2002. Clin. Diagn. Lab. Immunol.
  9138-43.

MH49981
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SP Induces Activation and Replication of HIV

• Ho, et al 1996. AIDS Res. Hum. Retroviruses
  12195-99.
• Li, et al 2001. J. Neuroimmunol. 12167-75.
• Lai, et al 2001. Proc. Natl. Acad. Sci. USA.
  983970-75.
• Ho, et al 2002. FASEB J. 16616-18.

MH49981
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SP Receptor Antagonists Inhibit HIV-1
InfectionLai, et al 2001. Proc. Natl. Acad.
Sci. USA. 983970-75.
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Neurokinin-1 receptor (NK-1R) antagonists

• Peptide and Non-peptide
• NK-1R, dual, and pan-NK
• Classification (groups)
• Diamines
• Amino-ethers
• Perhydroisoindoles
• Benzylpiperidine amides
• Quinoline and napthyridine amides
• Trytophan analogues

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Neurokinin-1 receptor (NK-1R) antagonists
Peptide Spantide-1 (Peninsula)(1985) - Full
length SP analog Spantide-2,3 - Truncated
C-terminal analogs (6-8 aa)
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Neurokinin-1 receptor (NK-1R) antagonists
Non-Peptide CP-96,345 (1991) Pfizer
quinuclidine residue - benzhydryl double-ring
motif CJ-11,974 Pfizer improved
bioavailability RP-67580 perhydroisoindole
family L733060 Merck piperidine Aprepitant
(Emend) MK-869 morpholine acetyl - benzylic
methyl group fluorine - decreases oxidative
metabolism
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Inhibition of HIV infection of human MDM by NK-1R
antagonists
Antagonist Source HIV inhibition
(RT) RP-67,580 RPR 10-6 M 72 10-7 M
78 10-8 M 79 CP-96,345 Pfizer 10-6
M 54 10-7 M 43 10-8 M
NE Spantide Peninsula, Inc. 10-5 M
27 10-6 M 50 10-7 M 73
To be moved to Project 1 or 2
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Inhibition of HIV infection of human MDM by NK-1R
antagonists
Antagonist Source HIV inhibition
(RT) L733060 Merck 10-6 M 54.6 10-7
M 44 L733061 (enantiomer) Merck 10-6 M
NE 10-7 M NE LY309809/LY307679 (inact.)
Lilly R-113381, NK-1-3 antagonist Sankyo Co.
Ltd. 10-6 M 79.8 10-7 M
69.6 10-8 M 45.8 Aprepitant
To be moved to Project 1 or 2
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Rationale for Clinical Development of Substance P
Antagonists in Depression

• Substance P (SP) is released in response to
  stressful stimuli
• SP is localized in regions controlling affective
  behavior
• (colocalization with select NA and 5-HT neurons
  in specific
• pathways)
• Amygdala
• Limbic areas
• Monoamine nuclei (raphe, locus ceruleus)
• Hypothalamus
• SP binding to NK-1 receptors elicits mood and
  emotional
• changes

Kramer MS et al. Science 2811640, 1998
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Structural Formula of Emend (Aprepitant) Empirical
formula C23H21F7N4O3
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SP receptor antagonist (Emend) potently inhibits
HIV (Bal) replication of human macrophages
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Overview
Projects 1 and 2 - Basic Projects 1. Will
determine in vitro efficacy, antiviral
specificity 2. Will determine cellular mechanism
of action and specificity 3. With private sector
partner (Boston Biomedica,Inc., Dr. Janet
Lathey), will determine synergy with known HIV
antivirals 4. Will guide selection of optimal
neurokinin-1R antagonist for preclinical/clinical
a. Specificity b. Potency c. Toxicity
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Overview
Project 3 - Pre-clinical Non-Human Primate
Project 1. Will determine safety and
pharmacokinetics 2. Will determine anti-SIV
activity 3. Will evaluate effects on behavior
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Overview
Project 4 - Human Studies of Neurokinin-1
Receptor (NK-1R) Antagonists in HIV-1 1. Will
determine safety 2. Will determine antiviral HIV
activity Phase 1B clinical trial 3. Will
determine pharmacokinetics and viral dynamics 4.
Will evaluate immunomodulatory effects 5. Will
evaluate depressive behavior effects
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Safety issues to be addressed (Aprepitant and
other NK-1R antagonists)

• Viral resistance/escape
• - Projects 1, 3, 4 Core B
• Increased SIV/HIV viral burden
• - Projects 3, 4
• Immunosuppression impaired immune modulation
• - Projects 2, 3, 4
• Behavioral changes
• - Projects 3, 4
• Negative interaction with retrovirals
• - Projects 3 Core B

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Interactions Between Projects
Cellular Mechanism
P2
P1
Antiviral
Antiviral
Antiviral
Immunomodulatory
Immunomodulatory
P4
P3
Safety
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PI and CI R01 List
Douglas R01 MH49981, R01 AA13547, U01
AI32921 Ho R01 DA12815, R21 DA16022 Kilpatrick
R01 GM64552 Lackner R01 NS30769, R01
MH61192, R01 DK50550 Tebas U01 AI32783 Evans
R01 MH44618, R25 MH60490, R01 MH067501 Cnaan
P01 CA097323, U01 NS045803 Barrett U01 Orange
K08 AI055602
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