Gorilla Adenovirus Vectors for

Molecular Therapeutics and Vaccines

ASGCT 18th Annual Meeting May 13 , 2015

Adenovectors and Packaging Cell Lines

GenVec Technology

• Adenovectors with superior performance characteristics for therapeutics and vaccines

• Broad spectrum of applications for the platform

• Significant vector construction and manufacturing expertise

• Additional viral gene deletions enhance safety and provide large packaging capacity

• Proprietary cell lines supported by FDA master file

1

Human and Nonhuman Types

GenVec Adenovectors

• Multiple adenovirus vector types

Human

Monkey

Chimpanzee

Gorilla

• Gorilla adenovirus (GC44, GC45, GC46)

New adenovirus serotypes isolated from wild gorilla

Similar to species C human adenovirus

Grow productively in our cells lines

Very low seroprevalence in human populations

2

Human Seroprevalence

Gorilla Adenovectors

• Analyzed gorilla adenovirus vectors for prevalence in human populations

US

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)

3

The U.S. Population Does Not Have High Levels of GC46-specific Neutralizing Antibodies

Approximately 240 serum samples were tested for neutralization of GC46

Frequency Titer distribution

IC90 titer

> 200 16-200 ≤ 16

GC46 SAV7 Ad5

100

1000

Tite

r (I

C-9

0)

200

Seropositives were infrequent, with titers too low to be inhibitory (IC90 = 200)

GC46 5.5%

94.5%

Ad5

42.5%

36.1% 21.4%

57.3%

41.3%

1.3%

SAV7

4

Distinct Advantages for Molecular Vaccines

Gorilla Adenovirus Vectors

Promising results in several vaccine preclinical models

• High-level, durable antibody responses from single administration

• High-level, durable T cell responses from single administration

• Repeat administration boosts responses

5

RSV Data Summary

• Protective – upper and lower respiratory tracts

• Durable – at least six months of protection

• Rapid – protection within two weeks

• Safe – no vaccine enhanced disease

• Breadth – immunological coverage of RSV diversity

6

Comparative Immunogenicity: Neutralizing Activity and Protection

4 Weeks

Serum for

neutralizing

antibody titer

RSV, 106 PFU, i.n. Buffer (FFB) rAd5F.0, 107 or 109 pu SAV7F.0, 107 or 109 pu GC46F.0, 107 or 109 pu

F Antigen IM

0

Lung RSV

5 days

post-

challenge

BALB/c H-2d

d

7

Comparative Immunogenicity

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)

Protection against RSV Serum neutralizing activity

• 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

* = significantly less than RSV-primed mice

Single Administration

8

Time Course of Pulmonary RSV Replication in Immunized Mice

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

* = 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

9

Characterization of GC46 Immunogenicity in the Cotton Rat Model

8 Weeks

Serum

RSV Long, 105 PFU, i.n. Buffer (FFB)

FI-RSV (Lot 100) 1:125 GC46F.0, 106 or 109

Lung RSV Nose RSV Histopathology

5 Days

post-

challenge

0 4

FI-RSV (Lot 100) 1:125

Serum

10

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)

GC46.F0 Immunogenicity in the Cotton Rat Model

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

RSV FFB FI-RSV RSV FFB 106 109

GC46.F0

(pu)

Lung

LOD

Nasal

LOD FI-RSV

11

Durable Neutralizing Antibody Titers Induced by GC46.F0

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

12

Upper and Lower Respiratory Tract Protection At Six Months

Lung

0

50

100

150

200

250

300

350

400

RSV A2 FI-RSV GC46.F0 e7GC46.F0 e9

Titer

(PF

U /

gra

m lung)

RSV 107 109

GC46.F0 FI-RSV

RSV A2 e6 FI-RSV 1:100 GC46.F0 e7 GC46.F0 e9100

1,000

10,000

100,000

RS

V t

iter

(PF

U /

nose)

Nasal

RSV 107 109

GC46.F0 FI-RSV

13

Malaria Data Summary

• Robust antigen specific T cell responses

• Robust antigen specific antibody responses

• Protection against P. yoelli challenge

14

%

C

SP

+ I

FNg+

CD

8+ T

-Cells

0

2

4

6

8

10

12

AdNull Ad5 GC44 GC45 GC46

Single administration of GC vector expressing PyCSP induces robust antigen specific T cell responses in mice

p

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

Comparative Immunogenicity Antigen Specific T Cell Dose Response

Single administration of GC vector expressing PyCSP induces robust antigen

specific T cell responses in mice

16

P<

0.5

Ad

Nu

ll

GC

46

Ad5

Na

ive

1 x 107 1 x 109

1 x 108

AdN

ull

GC

46

Ad5

AdN

ull

GC

46

Ad5

Comparative Immunogenicity Antigen Specific Antibody Dose Response

Single administration of GC vector expressing PyCSP induces robust

antigen specific antibody responses in mice

17

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

Comparative Immunogenicity P. yoelii Protection Against Challenge

18

HSV Data Summary

• Rapid, durable reduction in clinical symptoms HSV2 challenge in ovariectomized female mouse

HSV2 recurrence model in female guinea pigs

• Rapid viral load reduction HSV2 challenge in ovariectomized female mouse

• Robust, durable T cell response

• Repeat administration boosts T cell response

19

Schematic of HSV2 Challenge Model Using Ovariectomized Mice

Single I.M. Injection

20

Reduced Clinical Symptoms

No treatment Blend of GV46.UL19 and GC46.UL47, 1e9 pu each

21

Reduced Viral Load

qPCR assay Plaque assay

p< 0.01 p< 0.05

No treatment Blend of GV46.UL19 and GC47, 1e9 pu each

22

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

Robust T Cell Response and Viral Load Reduction

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

23

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 Administration

Repeat immunization with gorilla adenovirus vectors boosts antigen specific T cell responses

24

Schematic of HSV2 Recurrence Model Using Female Guinea Pigs

25

Rapid, Durable Reduction of Clinical Symptoms for at Least 63 Days

Incidence

p

p

• Adenovectors with superior performance characteristics for therapeutics and vaccines

• Broad spectrum of applications for the platform

• Grow to high titer on our cells lines

• Support additional viral gene deletions for enhanced safety and large packaging capacity

• Promising results in several vaccine preclinical models High-level, durable antibody responses from single administration

High-level, durable T cell responses from single administration

Repeat administration boosts responses

Gorilla Adenovectors

28

Acknowledgments

Mike Cranfield

Keith Limbach

Noelle Patterson

Maureen Stefaniak

Eileen Villasante

Tom Richie

Duncan McVey

Jason Gall

Teresa Johnson

Lisa Wei

Joe Bruder

Chris Lazarski

Ping Chen

Damodar Ettyreddy

Barney Graham

Holly Torano

Hubert Kuete

Andrew Glenn

David Rangel

Grace Lee

Randy Osborn

Johanna Harvel

29