Gorilla Adenovirus Vectors For Molecular Therapeutics and …content.equisolve.net/genvec/media/3934040899bee60a9549c... · 2015. 11. 4. · Analyzed gorilla adenovirus vectors for

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

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

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


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


U.S. Population Low Levels of GC46-specific Neutralizing Antibodies

4

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

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


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

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


RSV Neutralizing Antibody (NAb) titer

Neutralizing Antibodies Titers Induced by GC46.F0

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


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

*

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


(LOD = Limit

of Detection)

Durable Antibody Titers


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


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

Effective Immune Response


A B C D

Malaria Parasite Transmission Blocked in Mosquito

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


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



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


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


HSV Viral Load Reduced

Robust Immune Response

17


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

Reduced Clinical Symptoms

18


HSV Challenge, Untreated HSV Challenge, Treated with Blend of GV46.UL19 and GC46.UL47, 1e9 pu each

Repeat Administration

19

Ad Prime

Ad Boost -

GC45

1E9

-

GC45

1E7 FFB

FFB

GC45

1E7

GC45

1E7

-

GC45

1E9

GC45

1E9

GC45

1E9

Ad Prime

Ad Boost -

GC45

1E9

-

GC45

1E7 FFB

FFB

GC45

1E7

GC45

1E7

-

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


Immune Response Increased

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


Acknowledgements

21

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


Patrick Duffy

Charles Anderson

Kelly Rausch

Shaji Daniel

Olga Muratova

Holly Torano

Vaccine

Research

Center

www.genvec.com

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Gorilla Adenovirus Vectors For Molecular Therapeutics and …content.equisolve.net/genvec/media/3934040899bee60a9549c... · 2015. 11. 4. · Analyzed gorilla adenovirus vectors for