Neurokinin-1 Receptor Antagonist (Aprepitant) Inhibits Drug-Resistant HIV-1 Infection of Monocyte-Derived Macrophages in vitro Xu Wang, Steven D. Douglas, Jian-Ping Lai, Pablo Tebas, Janet L. Lathey, and Wen-Zhe Ho
Dec 30, 2015
Neurokinin-1 Receptor Antagonist (Aprepitant) Inhibits Drug-Resistant HIV-1
Infection of Monocyte-Derived Macrophages in vitro
Xu Wang, Steven D. Douglas, Jian-Ping Lai, Pablo Tebas, Janet L. Lathey, and Wen-Zhe Ho
Research Aim
To investigate the role of Neurokinin-1 Receptor Antagonist (Aprepitant) in Drug-Resistant HIV-1 Infection.
Background
• Respite of the success of antiretroviral therapy in maintaining control of HIV viral replication, treatment failure occurs in more than 50% of the individuals on HAART therapy.
• Cellular targets are attractive because it is more difficult for HIV to develop resistance.
• We have shown that NK-1R antagonist (CP-96,345) inhibits HIV-1 infection of MDM in vitro by down regulating CCR5 expression. (PNAS, 98: 3970-3975,2001)
Aprepitant
Chemical Structure of Aprepitant
Aprepitant: FDA approved drug for the treatment of chemotherapy
induced nausea
Experimental DesignMonocyte/Macrophage from normal subjects
With or without the NK-1R antagonists
treat for 2 h
Different types of HIV strains infection
HIV p24 assay at day 0, 3,5 7 post-infection
HIV RT assay at day 4, 8,12 post-infection
Inhibition of HIV (Bal) Infection of MDM by NK-1R Antagonists (10-6 M)
0
5
10
15
20
25
30
Control
Aprepita
nt
CJ12,2
55
CP-96,
345
RP 67,5
80
L-733
060
HIV
RT
Act
ivit
y (1
03 c
pm
)
Effect of Aprepitant on HIV Infection
RT Activity (% of Contriol) R5 R5X4 X4
Dose Bal SF162 89.6 UG024
Aprepitant 10-6 M 17.2 14.3 36.9 89.1
Aprepitant 10-7 M 13.8 16.6 44.1 93.1
Aprepitant 10-8 M 57.6 60.1 52.4 89.9
Control 100.0 100 100.0 100
Virus
RT Activity (% of Contriol) R5 R5X4 X4
Dose Bal SF162 89.6 UG024
Aprepitant 10-6 M 17.2 14.3 36.9 89.1
Aprepitant 10-7 M 13.8 16.6 44.1 93.1
Aprepitant 10-8 M 57.6 60.1 52.4 89.9
Control 100.0 100 100.0 100
Virus
Dose-dependent Inhibition of Aprepitant on HIV Bal infection of MDM
0
50
100
150
0 200 400 600 800 1000
Aprepitant (nM)
HIV
RT
Act
ivit
y (
% o
f C
on
tro
l)
Effect of Aprepitant on AZT Resistant Virus Infection of MDM
0
4
8
12
16
20
24
HIV
P24
Lev
el (
x10
3 p
g/m
l) Aprepitant
Control
AZT Resistant HIV-1 (A012 G691-6)
Day 0 Day 3 Day 5 Day70
50
100
150
200
250
300
HIV
P2
4 L
ev
el (
pg
/ml)
Aprepitant
Control
AZT Resistant HIV-1 (A018 G901-6)
Day 0 Day 3 Day 5 Day7
A012 G691-6 and A018G901-6 were kind gifts from NIH AIDS Research and Reagent Program
Effect of Aprepitant on RT Inhibitor-Resistant HIV Infection of MDM
0
100
200
300
400
500
HIV
P24
Lev
el (p
g/m
l)
Aprepitant
Control
Day 0 Day 3 Day 5 Day7
RT Inhibitor-Resistance HIV-1 TC 60
B
0
5000
10000
15000
20000
25000
30000
HIV
P24
Lev
el (p
g/m
l)
Aprepitant
Control
Day 0 Day 3 Day 5 Day7
RT Inhibitor-Resistance HIV-1 TC49
A
TC49 and TC60 (R5 HIV strain) were kind gifts from Dr. Besty Johnston White and Dr. David Katzenstein (Stanford University)
Effect of Aprepitant on HIV co-receptor(CCR5) Expression in MDM
0
20
40
60
80
100
120
140
CC
R5
mR
NA
Lev
el (
% o
f C
on
tro
l)
Aprepitant Control
Exp1 Exp 2 Exp 3 Exp 4
0
20
40
60
80
100
120
140
CD
4+C
CC
R5+
cel
ls (
% o
f C
on
tro
l) Aprepitant Control
Exp1 Exp 2 Exp 3 Exp 4
Enhancing Effect of Aprepitant on anti-HIV Drug-mediated HIV Inhibition in MDM
0
5
10
15
20
25
HIV
RT
(1
03 c
pm
)
Aprepitant 10-6M
AZT 10-11M
Efavirenz 10-10M
Indinavir10-15M
----
+
---
-+
--
+
+
--
--+
-
+
-+
-
---+
+
--+
Summary 1
• Among the NK-1R antagonists examined
Aprepitant had a better inhibitory effect on
HIV infection of MDM.
• Aprepitant, in a dose-dependent manner,
inhibits HIV R5 strain infection, partially
inhibited R5X4 strain infection, and had little
effect on HIV X4 strain infection.
Summary 2
• Aprepitant potently inhibited the AZT-
resistant strains (A018 G901-6 and A012
G691-6) infection of MDM.
• Aprepitant also suppressed RT inhibitor
resistant strains (TC 49 and TC 60) infection
of MDM.
Summary 3
• Aprepitant down-regulated CCR5, the co-
receptor for HIV entry into MDM.
• Aprepitant enhanced the anti-HIV activity of
antiretrovirals (AZT, efavirenz, and
indinavir) in MDM.
Conclusion
Our data suggest that NK-1R antagonists merit further investigation as potential HIV therapeutic agents
• This investigation was supported by MH 076388 and MH 049981 from NIH
Acknowledgments
• Jian-Ping Lai• Li Song• Wenzhe Ho
• Pablo Tebas
Seracare BioServices Janet L. Lathey
• Donald E. Campbell • Eric Riedel• Steven D. Douglas
NK-1R Antagonists: Anti-HIVMechanisms
Project 1PI: Wenzhe Ho
NK-1R Antagonist
Anti-HIV Mechanisms
PBMC from Normal Subjects Drug ResistantSynergistic Studies
PBMC from HIV-infected Subjects
HIV Activation SP Induction
Microglia
HIV Replication Cytokine Production
Neuronal Cells (NT-2N)
HIV gp120 HIV Tat
Schematic Diagram of Project 1
HIV Replication (Direct Effect) CCR5 (Indirect Effect) SP (Indirect Effect)
Aims 1 & 2 Aims 2 & 3 Aim 4 Aim 4
Aim 1a: To determine whether the NK-1Rantagonists inhibit HIV infection of PBMC
M-, T-, and dual tropic HIV strains
HIV-1 p24 assay at day 7 post-infection (Core B)
With or without the NK-1R antagonists
PBMC from normal subjects
Cytotoxicity assay
Table 1 – Drug Resistant Isolates to be used
Isolates Biotype Resistance
139-1 SI (R5X4) AZT
HIV-174V/MT2 X4 DdI, ddC
HIV-1 LAI-M184V
SI 3TC
N119 SI (X4) Nevirapine
J302 SI Saquinavir
PBMC from Normal Subjects
Antiretroviral drugs (RT and protease inhibitors)
and/or
HIV p24 (Core B)
M- and T-tropic strains
w or w/o NK-1R antagonists
w or w/o NK-1R antagonists
Drug resistant strains
Aim 1b – To determine the effect of NK-1R antagonists on drug-resistant isolates and the synergistic effect of the antagonists
Table 2 – Antiretroviral Drugs to be Used
Drug Classification
ZidovudineDDC
EfavirenzLopinavirRitonavir
T20PA-457
RT Inhibitor (NRTI)RT Inhibitor (NRTI)
RT Inhibitor (NNRTI)Protease InhibitorProtease InhibitorFusion Inhibitor
Maturation Inhibitor