gorilla adenovirus vectors for molecular therapeutics and...
TRANSCRIPT
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Developing a pipeline of innovative therapeutics and vaccines that deliver on the promise of gene-based medicine
Gorilla Adenovirus Vectors
For Molecular Therapeutics and Vaccines Douglas E. Brough, Ph.D.
Chief Scientific Officer
Vaccines R&D Conference
November 2015
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Adenovectors with superior performance characteristics for therapeutics and vaccines
Broad spectrum of applications for the platform
Significant vector construction and manufacturing experience
Additional viral gene deletions enhance safety and provide large packaging capacity
Proprietary cell lines supported by FDA master file
AdenoVerse™ Technology Adenovectors and Packaging Cell Lines
1 November 2015 Vaccines R&D Conference
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Multiple adenovirus vector types
Human
Monkey
Gorilla
Gorilla adenovirus (GC44, GC45, GC46)
New adenovirus serotypes isolated from wild gorilla
Similar to species C human adenovirus
Grow productively in our cell lines
Very low seroprevalence in human populations
AdenoVerse™ Vectors Human and Nonhuman Types
2 November 2015 Vaccines R&D Conference
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Analyzed gorilla adenovirus vectors for prevalence in human populations
U.S.
Sub-Saharan Africa
Benchmarked against Ad5, SAV7, Ad35
Seropositives in the human population were infrequent
The few positive titers were found to be very low
Too low to be inhibitory (IC90-200)
Gorilla Adenovectors Human Seroprevalence
3 November 2015 Vaccines R&D Conference
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U.S. Population Low Levels of GC46-specific Neutralizing Antibodies
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Approximately 240 serum samples were tested for neutralization of GC46
Seropositives were infrequent, with titers too low to be inhibitory (IC90 = 200)
Frequency Titer distribution
IC90 titer
> 200 16-200 ≤ 16
GC46 SAV7 Ad5
100
1000
Tite
r (I
C-9
0)
200
GC46 5.5%
94.5%
Ad5
42.5%
36.1% 21.4%
57.3%
41.3%
1.3%
SAV7
November 2015 Vaccines R&D Conference
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Promising results in several vaccine preclinical models
High-level, durable antibody and T cell responses from single administration
Repeat administration boosts response
Gorilla Adenovirus Vectors Distinct Advantages for Molecular Vaccines
5 November 2015 Vaccines R&D Conference
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Comparative Immunogenicity Single Administration
6
* = significantly less than RSV-primed mice
GC44.F0 and GC46.F0 induced titers comparable to an immunization with 1 x 106 pfu of RSV
Can differentiate performance based on low dose immunizations November 2015 Vaccines R&D Conference
101
102
103
104
105
106
107
RSV
SAV7.F0 GC46.F0
109
107
PF
U/g
ram
lung
FFB 109
107 10
910
7
Ad5.F0
104
103
102
101
106
105
107
107 109
GC46.F0 107 109
SAV7.F0 107 109
Ad5.F0
64
128
256
512
PR
NT
(IC
-50)
RSV Neut Ab titer Protection Against RSV Challenge
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Durable Immune Response
7
16
32
64
128
256
512
1024
0 2 4 6 8 10 12 14 16 18 20 22 24 26
PR
NT
titer
(IC
50
)
Weeks post-immunization
GC46.F0 109 pu
GC46.F0 107 pu
FI-RSV
November 2015 Vaccines R&D Conference
RSV Neutralizing Antibody (NAb) titer
Neutralizing Antibodies Titers Induced by GC46.F0
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Protective Immunity Pulmonary RSV Replication in Immunized Mice
8
* = significantly different than FFB-primed mice
Mice, Balb/c
Challenge at 4 weeks post-immunization
GC46.F0 108 pu
RSV challenge 5 x 106 PFU
November 2015 Vaccines R&D Conference
1 2 3 4 510
1
102
103
104
105
106
107
RS
V titers
(P
FU
/ g
lung)
Day post-challenge
FFB
RSV
GC46.F0
*
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Protective Immunity Cotton Rat Model
9
4
5
6
7
8
9
10
RSV FFB FI-RSVGC46 F 1e6 puGC46 F 1e9 pu
Ge
oM
ea
n L
og
2 T
ite
rs (
IC-6
0)
Day0 Day 28 Day 56
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
RSV FFB FI-RSVGC46 F 1e6 puGC46 F 1e9 pu
Ge
oM
ea
n L
og
10
Tite
rs (
pfu
/g)
Lung
Nasal
109 106
GC46.F0
(pu)
RSV FFB FI-RSV RSV FFB 106 109
GC46.F0
(pu)
Lung
LOD
Nasal
LOD FI-RSV
Serum neutralizing antibody Protection against RSV
Lung and nasal titers were reduced to undetectable
Nasal RSV titers were reduced 10-fold with GC46.F0 106 pu immunization
November 2015 Vaccines R&D Conference
(LOD = Limit
of Detection)
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Durable Antibody Titers
10 November 2015 Vaccines R&D Conference
0 2 8 4 2 9 8 1 2 6 10 2
10 3
10 4
10 5
d a y s
E L I
S A
u n
i t s
Anti Pfs 230 titer GC25+GC230 Single dose GC Prime / Protein Boost Protein Prime / GC Boost
Protein Prime / Protein Boost
Ab Titers Induced by GC46.25 + GC46.230
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11 November 2015 Vaccines R&D Conference
GC25+GC230 Single dose
GC Prime / Protein Boost
Protein Prime / GC Boost
Protein Prime / Protein Boost
0 2 8 4 2 9 8 1 2 6 10 2
10 3
10 4
10 5
d a y s
E L I
S A
u n
i t s
Anti Pfs 25 titer
Durable Antibody Titers Ab Titers Induced by GC46.25 + GC46.230
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Effective Immune Response
12 November 2015 Vaccines R&D Conference
A B C D
Malaria Parasite Transmission Blocked in Mosquito
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CD8+ T Cell, Antibody and Protection
Comparative Immunogenicity
Single administration of GC vector expressing PyCSP induces
robust antigen-specific T cell responses in mice
13
%
C
SP
+ I
FNg+
CD
8+ T
-Cells
0
2
4
6
8
10
12
AdNull Ad5 GC44 GC45 GC46
p
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Comparative Immunogenicity
14
Antigen-Specific T Cell Dose Response
0
2
4
6
8
10
12 Media HA332-340
PyCSP280-288
PyCSP57-70
AdN
ull
GC
46
Ad5
Naiv
e
1 x 107 1 x 109
1 x 108
P< 0.001
P< 0.001
AdN
ull
GC
46
Ad5
AdN
ull
GC
46
Ad5
%
C
SP
+ I
FNg+
CD
8+ T
-Cells
Single administration of GC vector expressing PyCSP induces robust antigen-specific
T cell responses in mice
November 2015 Vaccines R&D Conference
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Comparative Immunogenicity
15
P. Yoelii Protection Against Challenge
0
10
20
30
40
50
60
DNA Ad5
DNA GC44
DNA GC45
DNA GC46
DNA null GC46 null
Naïve
% P
rote
ctio
n 36%
43%
14%
50%
0% 0%
Prime Boost
November 2015 Vaccines R&D Conference
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Robust T Cell Response
16
n=18/Group
0
50
100
150
200
Perc
ent
of
Naï
ve V
iral
Tit
er
Naïve UL19 UL19 + UL47
Viral Load is Reduced (Plaque)
n=6/Group
T- Cell Response is Enhanced
Day 0 Day 14
Single I.M. Injection Splenocyte Harvest
Day 0 Day 21
Vaginal Swab(Day +7) Single I.M. Injection
Day 14
HSV2 Infection
0
2
4
6
8
10
12
14
16
18
20
Naïve UL19 UL19+ UL47
Perc
ent
CD
8+
IFNg
+ T
Cel
ls
November 2015 Vaccines R&D Conference
HSV Viral Load Reduced
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Robust Immune Response
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November 2015 Vaccines R&D Conference
qPCR assay Plaque assay
Post HSV Challenge – Untreated vs. Treatment with blend of GC46.UL19 and GC46.UL47, 1e9 pu each
Untreated
Treated
Untreated
Treated
HSV Viral Load Suppressed
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Reduced Clinical Symptoms
18
November 2015 Vaccines R&D Conference
HSV Challenge, Untreated HSV Challenge, Treated with Blend of GV46.UL19 and GC46.UL47, 1e9 pu each
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Repeat Administration
19
Ad Prime
Ad Boost -
GC45
1E9
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GC45
1E7 FFB
FFB
GC45
1E7
GC45
1E7
-
GC45
1E9
GC45
1E9
GC45
1E9
Ad Prime
Ad Boost -
GC45
1E9
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GC45
1E7 FFB
FFB
GC45
1E7
GC45
1E7
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GC45
1E9
GC45
1E9
GC45
1E9
4 week
prime/boost
interval
12 week
prime/boost
interval
** *** **** A) B)
Repeat immunization with gorilla adenovirus vectors boosts antigen specific T cell responses
November 2015 Vaccines R&D Conference
Immune Response Increased
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AdenoVerse™ Vectors
Adenovectors with superior performance characteristics for therapeutics and vaccines
Broad spectrum of applications for the platform
Grow to high titer on our cell lines
Support additional viral gene deletions for enhanced safety and large packaging capacity
Gorilla vectors show promising results in several vaccine preclinical models
High-level, durable antibody responses from a single administration
High-level, durable T cell responses from a single administration
Repeat administration boosts responses
20 November 2015 Vaccines R&D Conference
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Acknowledgements
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Mike Cranfield
Keith Limbach
Noelle Patterson
Maureen Stefaniak
Eileen Villasante
Tom Richie
Duncan McVey, Jason Gall, Teresa Johnson, Lisa Wei, Chris Lazarski, Ping Chen, Holly Torano, Hubert Kuete, Andrew Glenn, David Rangel, Grace Lee, Randy Osborn, Johanna Harvel
Damodar Ettyreddy, Allison Keene, Shanyi Jang, Bryan Butman, Joe Bruder
Barney Graham
November 2015 Vaccines R&D Conference
Patrick Duffy
Charles Anderson
Kelly Rausch
Shaji Daniel
Olga Muratova
Holly Torano
Vaccine
Research
Center
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www.genvec.com