Download Dr. Christopher Thanos - Halozyme Therapeutics

American Association for Cancer Research
Investor Meeting
Targeting the Tumor
Microenvironment
Copyright © 2016 Halozyme, Inc.
Forward-Looking Statements
All of the statements in this presentation that are not statements of historical
facts constitute forward-looking statements within the meaning of the Private
Securities Litigation Reform Act of 1995. Examples of such statements include
future product development and regulatory events and goals, anticipated
clinical trial results and strategies, product collaborations, our business
intentions and financial estimates and results. These statements are based
upon management’s current plans and expectations and are subject to a
number of risks and uncertainties which could cause actual results to differ
materially from such statements. A discussion of the risks and uncertainties
that can affect these statements is set forth in the Company’s annual and
quarterly reports filed from time to time with the Securities and Exchange
Commission under the heading “Risk Factors.” The Company disclaims any
intention or obligation to revise or update any forward-looking statements,
whether as a result of new information, future events, or otherwise.
1
American Association for Cancer Research
Investor Meeting
Opening Remarks
Dr. Helen Torley
President and CEO
April 18, 2016
Copyright © 2016 Halozyme, Inc.
Agenda
Time
Topic
5 minutes
Opening Remarks
10
minutes
Focus on the Tumor Microenvironment
10
minutes
10
minutes
PEGPH20 Immuno-Oncology
PEG-ADA2: PEGylated Adenosine Deaminase 2
10
minutes
HTI-1511: Anti-EGFR Antibody-Drug Conjugate
15
minutes
Questions and Answers
Presenter
Dr. Helen Torley
President and CEO
Dr. Michael LaBarre
VP, Chief Scientific Officer
Dr. Sanna Rosengren
Director, Immunology and
Cell Biology
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
All
3
Agenda
Time
Topic
5 minutes
Opening Remarks
10
minutes
Focus on the Tumor Microenvironment
10
minutes
10
minutes
PEGPH20 Immuno-Oncology
PEG-ADA2: PEGylated Adenosine Deaminase 2
10
minutes
HTI-1511: Anti-EGFR Antibody-Drug Conjugate
15
minutes
Questions and Answers
Presenter
Dr. Helen Torley
President and CEO
Dr. Michael LaBarre
VP, Chief Scientific Officer
Dr. Sanna Rosengren
Director, Immunology and
Cell Biology
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
All
4
American Association for Cancer Research
Investor Meeting
Focus on the Tumor
Microenvironment
Dr. Michael LaBarre
Vice President, Chief Scientific Officer
April 18, 2016
Our Focus: Developing Therapeutics That Target
the Tumor Microenvironment (TME)
Modify the TME Structure
• PEGylated human recombinant PH20 (PEGPH20)
– to deplete hyaluronan (HA) in the TME and increase tumor access
Leverage the TME Biochemistry and Physiology
• PEGylated adenosine deaminase 2 (PEG-ADA2)
– to deplete adenosine (an immune checkpoint) in the TME
• Anti-EGFR-ADC (HTI-1511)
– to bind preferentially to EGFR at low pH in TME and deliver cytotoxic
drug conjugate to tumor cells
6
The Tumor Microenvironment
Cancer cell
Vasculature
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
7
The Tumor Microenvironment
Cancer cell
Vasculature
HA and Collagen
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
8
The Tumor Microenvironment
Cancer cell
Vasculature
HA
Immune Cells
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
9
The Tumor Microenvironment
Cancer cell
Vasculature
HA
Adenosine and
Molecular Receptors
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
10
The Tumor Microenvironment
Cancer cell
Regions of
- Low O2
- Low pH
Vasculature
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
11
The Tumor Microenvironment
Cancer cell
Vasculature
HA and Collagen
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
12
Hyaluronan (HA) Can Be a Barrier to Therapeutic
and Immune Cell Access to Tumor Cells
• HA is a Structural Carbohydrate
HA Surrounding a Cancer Cell
Red Blood Cells
– Hydrophilic, viscous polysaccharide
• Stabilizes the Tumor Microenvironment
– Provides structure for extracellular and
cell surface components
HA (Red) surrounding a single breast cancer
cell overexpressing HAS3 (Bright Green)
Kultti, et al. JBC. 2006,281:15821
Brekken et al. Anticancer Res. 20:3503 (2000)
Provenzano and Hingorani, Br J Cancer 108:1 (2013)
3 Thompson et al. Mol Cancer Ther. 9:3052 (2010)
4 Stylianopoulos et al. PNAS. 110:18632 (2013)
5 Singha et al. . Mol Cancer Ther. 14:523 (2015)
1
2
13
Hyaluronan (HA) Can Be a Barrier to Therapeutic
and Immune Cell Access to Tumor Cells
• HA is a Structural Carbohydrate
HA Surrounding a Cancer Cell
Red Blood Cells
– Hydrophilic, viscous polysaccharide
• Stabilizes the Tumor Microenvironment
– Provides structure for extracellular and
cell surface components
• Compromises Access to the Tumor
– Increased tumor interstitial pressure1,2
– Vasculature compression3,4
– Can decrease therapeutic and immune
cell access5
HA (Red) surrounding a single breast cancer
cell overexpressing HAS3 (Bright Green)
Kultti, et al. JBC. 2006,281:15821
Brekken et al. Anticancer Res. 20:3503 (2000)
Provenzano and Hingorani, Br J Cancer 108:1 (2013)
3 Thompson et al. Mol Cancer Ther. 9:3052 (2010)
4 Stylianopoulos et al. PNAS. 110:18632 (2013)
5 Singha et al. . Mol Cancer Ther. 14:523 (2015)
1
2
14
HA Accumulation Associated With Decreased
Survival in Some Tumors in Clinical Trials
Pancreatic Ductal Adenocarcinoma1
HA-high
Median Survival: 9.3 months
HA-low
Median Survival: 24.3 months
Whatcott, et al. Clin Cancer Res. 21:15 (2015)
Setala, et al. Br J Cancer 79:1133 (1999)
3 Ropponen, et al. Cancer Res. 58:342 (1998)
4 Antilla, et al. Cancer Res. 60:150 (2000)
5 Pirinen, et al. Int J Cancer 95:12 (2001)
6 Auvinen, et al. Am J Pathol. 156:529 (2000)
7 Lipponen, et al. Eur J Cancer 37:849 (2001)
1
2
Negative correlation with survival also reported in
gastric2, colorectal3, ovarian4, non-small cell lung5,
metastatic breast6 and prostate cancers7
15
PEGPH20 Targets Hyaluronan in the TME
Removal of HA by PEGPH20 demonstrated in HA-high tumor animal models to:
Decrease
intratumoral
pressure
Decompress
vasculature
Increase
perfusion
Increase
access for
therapeutics
Increase
access for
immune cells
PEGPH20
16
PEGPH20 Decreased HA in an HA-high Wilms’
Tumor Animal Model
WT-CLS1/HAS3 HA-high peritibial Wilms’ tumor model
Vehicle
PEGPH20 37.5 µg/kg (HED)
HA Staining
Nuclear Staining
HA (brown) staining with HTI-601
(HA-specific binding probe developed at Halozyme)
Tumors harvested 6h after 2 doses of Vehicle
or PEGPH20 on days 0 and 3
Cowell et al. (2016). AACR Annual Meeting, Poster #2463
17
PEGPH20 Reduced Tumor Interstitial Fluid Pressure in
HA-high Prostate Cancer Animal Model
PC3 peritibial prostate tumor model
1 .2
N o r m a liz e d T u m o r IF P
PEGPH20 Dose (mg/kg)
1 .0
V e h ic le
0 .8
0 .0 1 5
~40 mmHg
0 .1 5
0 .6
IV dose
1 .5
0 .4
4 .5
0 .2
15
0 .0
-2 0
0
20
40
60
80
100
120
T im e a ft e r T r e a t m e n t ( m in )
Thompson et al. Mol Cancer Ther. 9:3052 (2010)
18
PEGPH20 Increased Tumor Perfusion in
HA-high Prostate Cancer Animal Model
PC3 peritibial prostate tumor model
Vehicle (24h)
PEGPH20 (24h)
Hyperechoic microbubbles imaged to visualize vasculature “space” or vascular
area of peritibial PC3 tumors ± PEGPH20 (15 mg/kg, IV). Blue tracing is tumor area.
Thompson et al. Mol Cancer Ther. 9:3052 (2010)
19
Vascular Decompression Increased Drug Delivery
in HA-high Prostate Cancer Animal Model
PC3 peritibial prostate tumor model
*
3 0
2 5
(n g /µ g D N A )
T u m o r L ip o s o m a l D o x o r u b ic in
3 5
Increased tumor DOXIL
(measured as doxorubicin)
324%↑
[doxorubicin]tum
* p < 0.05
2 0
1 5
1 0
5
0
L ip o s o m a l
D o x o r u b ic in
A lo n e
P EG P H 20
3h
L ip o s o m a l
D o x o r u b ic in
Increased drug accumulation not observed in normal tissues tested
Thompson et al. Mol Cancer Ther. 9:3052 (2010)
20
PEGPH20 Increased Activity of Vincristine and
Dactinomycin in HA-high Wilms’ Tumor Animal Model
1500
1250
V e h ic le
T u m o r V o lu m e , m m
3
(S E M )
WT-CLS1/HAS3 HA-high peritibial tumor model
1000
750
PEG PH 20
500
250
V in + D A C T
P E G P H 2 0 + V in + D A C T
0
0
7
14
21
D a y s o n S tu d y
PEGPH20 37.5 µg/kg (HED), vincristine 0.415 mg/kg,
dactinomycin 0.125 mg/kg 2x weekly
Cowell et al. (2016). AACR Annual Meeting, Poster #2463
21
PEGPH20 Increased Activity of Vincristine and
Dactinomycin in HA-high Wilms’ Tumor Animal Model
1500
1250
V e h ic le
T u m o r V o lu m e , m m
3
(S E M )
WT-CLS1/HAS3 HA-high peritibial tumor model
1000
750
PEG PH 20
500
250
Without PEGPH20
5/10 Regressions
V in + D A C T
P E G P H 2 0 + V in + D A C T
0
0
7
14
D a y s o n S tu d y
PEGPH20 37.5 µg/kg (HED), vincristine 0.415 mg/kg,
dactinomycin 0.125 mg/kg 2x weekly
21
With PEGPH20
10/10 Regressions
Cowell et al. (2016). AACR Annual Meeting, Poster #2463
22
PEGPH20 Increased Tumor Growth Inhibition of
shIDO-ST in HA-high Pancreatic Cancer Animal Model
KPC-derived pancreatic cancer model
300
lo g p h o to n s / s ( x 1 0
6
)
shIDO-ST:
Salmonella typhimurium (ST)
short hairpin RNA against
IDO (shIDO)
250
200
V e h ic le
150
PEG PH 20
s h ID O - S T
100
50
s h ID O - S T + P E G P H 2 0
0
0
10
20
30
40
p < 0 .0 1 ,
A N O V A
60
70
50
P o s t- tu m o r im p la n ta tio n ( d )
PEGPH20 2.25 mg/kg on day 13
shIDO-ST 5 x 106/dose on day 14, 15 and 16
(n=3/group)
Manuel et al. Cancer Immunol Res. 3:1096 (2015)
23
PEGPH20 With shIDO-ST Increased Immune Cell
Influx in HA-high Pancreatic Cancer Animal Model
KPC-derived pancreatic cancer model
PEGPH20 + Control-ST
PEGPH20 + shIDO-ST
Neutrophil
accumulation
Epi-fluorescence
Counts
PEGPH20 2.25 mg/kg on day 13
shIDO-ST 5 x 106/dose on day 14, 15 and 16
Tumors extracted 96 hours post-ST treatment
Manuel et al. Cancer Immunol Res. 3:1096, Figure S8 (2015)
24
PEGPH20 Increased Access of Various Drugs and
Immune Cells in HA-high Tumor Animal Models
Relative Size
larger
Leukocytes (e.g., NK cells1, T cells2, neutrophils3)
shIDO-ST cellular immunotherapy3
Liposomes/nanoparticles (e.g., ABRAXANE® 4, DOXIL® 5)
Monoclonal antibodies (e.g., cetuximab6, trastuzumab1)
Small molecules (e.g., gemcitabine7)
smaller
Singha et al. Mol Cancer Ther. 14:523 (2015), 2 Singha et al. AACR Natl Mtg(2015), 3 Manuel et al. Cancer Immunol Res. 3:1096 (2015),
Measured as paclitaxel in Osgood et al. AACR PDA Mtg.(2014), 5 Thompson et al. Mol Cancer Ther. 9:3052 (2010),
6 Kang et al. Halozyme new data to be submitted for publication, 7Jacobetz et al. GUT 62:112 (2013)
1
4
25
Agenda
Time
Topic
5 minutes
Opening Remarks
10
minutes
Focus on the Tumor Microenvironment
10
minutes
10
minutes
PEGPH20 Immuno-Oncology
PEG-ADA2: PEGylated Adenosine Deaminase 2
10
minutes
HTI-1511: Anti-EGFR Antibody-Drug Conjugate
15
minutes
Questions and Answers
Presenter
Dr. Helen Torley
President and CEO
Dr. Michael LaBarre
VP, Chief Scientific Officer
Dr. Sanna Rosengren
Director, Immunology and
Cell Biology
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
All
26
American Association for Cancer Research
Investor Meeting
PEGPH20 Immuno-Oncology
Dr. Sanna Rosengren
Director, Immunology and Cell Biology
April 18, 2016
The Tumor Microenvironment
Cancer cell
Vasculature
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
28
The Tumor Microenvironment
Cancer cell
Vasculature
Immune Cell Involvement
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
29
Anti-Tumor T Cells Face Immunosuppressive Barriers
in the TME
Tumor
cell
Anti-tumor
T cell
Antigen-presenting
dendritic cell
30
Anti-Tumor T Cells Face Immunosuppressive Barriers
in the TME
Legend
Blocking
interaction
Tumor
cell
PD-1 PD-L1
PD-L1
PD-1
CTLA4
Anti-tumor
T cell
Antigen-presenting
dendritic cell
Immune
checkpoints
31
Anti-Tumor T Cells Face Immunosuppressive Barriers
in the TME
Regulatory
T cell
Myeloid
suppressor cell
M2
macrophage
Immunosuppressive
cells
Legend
Blocking
interaction
Tumor
cell
PD-1 PD-L1
PD-L1
PD-1
CTLA4
Anti-tumor
T cell
Antigen-presenting
dendritic cell
Immune
checkpoints
32
Anti-Tumor T Cells Face Immunosuppressive Barriers
in the TME
Regulatory
T cell
Myeloid
suppressor cell
M2
macrophage
Immunosuppressive
cells
Legend
Blocking
interaction
Tumor
cell
PD-1 PD-L1
PD-L1
High
Levels
PD-1
CTLA4
Anti-tumor
T cell
Antigen-presenting
dendritic cell
Immune
checkpoints
Immunosuppressive
cytokines and
growth factors
GRADIENT
33
Some Biomarkers of Immunosuppression Increased in
HA-high Colon Cancer Animal Model
CT26/HAS3 HA-high syngeneic colon tumor model
IL10
(immune checkpoint)
(regulatory T cell marker)
4
4
p = 0.008
3
2
2
2
1
1
1
0
0
0
6
S
T
S
C
6
A
/H
2
6
T
C
C
T
2
2
T
C
2
6
C
A
6
/H
T
S
2
6
2
T
C
3
3
3
3
3
p = 0.002
A
p = 0.009
Gene expression
(Actb-normalized)
CTLA4
FoxP3
/H
4
(immunosuppressive
cytokine)
Other immunosuppressive genes, including PD-L1 and IDO, were not significantly different in this model
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
34
Does PEGPH20 Increase Activity of Anti-CTLA4
Antibodies in HA-high Tumors?
CTLA4 engagement blocks co-stimulation of activated T cells
Anti-CTLA4 antibodies can lower tumor immunosuppression
PEGPH20 might enhance their efficacy in HA-high tumors
Cancer
cell
PD-1 PD-L1
PD-L1
PD-1
CTLA4
Anti-tumor
T cell
Antigen-presenting
dendritic cell
35
PEGPH20 Effect on Anti-CTLA4 Activity in HA-high
and HA-low Colon Cancer Animal Models
CT26/HAS3 HA-high model
Is o ty p e c o n tr o l
PEG PH20
A n ti- C T L A 4
P E G P H 2 0 + a n ti- C T L A 4
2000
3
T u m o r V o lu m e ( m m ) ± S E M
2500
1500
1000
**
p ≤ 0.002
to anti-CTLA4
and PEGPH20
500
0
0
5
10
15
D a y o n S tu d y
PEGPH20 37.5 µg/kg (HED) biweekly, 24h prior to antiCTLA4 or IgG2b isotype control (4 mg/kg)
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
36
PEGPH20 Effect on Anti-CTLA4 Activity in HA-high
and HA-low Colon Cancer Animal Models
CT26/HAS3 HA-high model
CT26 HA-low model
Is o ty p e c o n tr o l
Is o ty p e c o n tr o l
PEG PH20
P E G P H 2 0 + a n ti- C T L A 4
A n ti- C T L A 4
P E G P H 2 0 + a n ti- C T L A 4
2000
3
T u m o r V o lu m e ( m m ) ± S E M
A n ti- C T L A 4
2000
PEG PH20
2500
3
T u m o r V o lu m e ( m m ) ± S E M
2500
1500
1000
**
p ≤ 0.002
to anti-CTLA4
and PEGPH20
500
0
Not
significant
1500
1000
500
0
0
5
10
15
D a y o n S tu d y
PEGPH20 37.5 µg/kg (HED) biweekly, 24h prior to antiCTLA4 or IgG2b isotype control (4 mg/kg)
0
5
10
15
D a y o n S tu d y
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
37
Does PEGPH20 Increase the Activity of
Anti-PD-1/PD-L1 Antibodies in HA-high Tumors?
PD-1/PD-L1 interaction sends signal of exhaustion to T cell
Anti-PD-1/PD-L1 antibodies can lower tumor immunosuppression
PEGPH20 might enhance their efficacy in HA-high tumors
Cancer
cell
PD-1 PD-L1
PD-L1
PD-1
CTLA4
Anti-tumor
T cell
Antigen-presenting
dendritic cell
38
PEGPH20 Increased Activity of Anti-PD-1 in HA-high
Pancreatic Cancer Animal Model
KPC-derived HA-high pancreatic tumor model
V e h ic le
PEG PH20
A n ti- P D - 1
P E G P H 2 0 / a n ti- P D - 1
2000
3
T u m o r V o lu m e ( m m ) ± S E M
2500
1500
1000
*
p < 0.02 to anti-PD-1
and PEGPH20
500
0
0
5
10
15
20
D a y o n S tu d y
PEGPH20 37.5 µg/kg (HED) 2x weekly,
24h prior to anti-PD-1 (0.5 mg/kg)
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
39
PEGPH20 Increased Activity of Anti-PD-L1 in HA-high
Pancreatic Cancer Animal Model
KPC-derived HA-high pancreatic tumor model
Is o t y p e c o n t r o l
P E G P H 2 0 / is o t y p e c o n t r o l
A n t i- P D -L 1
P E G P H 2 0 / a n t i- P D - L 1
2000
3
T u m o r V o lu m e ( m m ) ± S E M
2500
1500
1000
***
500
p < 0.0001 to anti-PD-L1
and PEGPH20
0
0
5
10
15
20
25
D a y o n S tu d y
PEGPH20 37.5 µg/kg (HED) 2x weekly, 24h prior
to anti-PD-L1 or IgG2b isotype control (2 mg/kg)
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
40
PEGPH20 Combined With Anti-PD-L1 Induced
Regressions in HA-high Pancreatic Cancer Animal Model
% C h a n g e T u m o r V o lu m e fro m b a s e lin e
KPC-derived HA-high pancreatic tumor model
1400
1200
1000
800
600
With PEGPH20
3/8 Regressions
400
200
0
-2 0 0
Is o ty p e
PEG PH20 /
c o n tr o l
Is o ty p e c o n tr o l
PEGPH20 37.5 µg/kg (HED) 2x weekly, 24h prior
to anti-PD-L1 or IgG2b isotype control (2 mg/kg)
A n ti- P D - L 1
PEG PH20 /
A n ti- P D - L 1
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
41
Human Equivalent Dose of PEGPH20 Reduced HA
in HA-high Pancreatic Cancer Animal Model
KPC-derived HA-high pancreatic tumor model
Pre-treatment
24 hours after PEGPH20
(37.5 µg/kg)
HA Staining
Nuclear Staining
HA (brown) staining with HTI-601
(HA-specific binding probe developed at Halozyme)
PEGPH20 37.5 µg/kg (HED) 2x weekly, 24h prior
to anti-PD-L1 or IgG2b isotype control (2 mg/kg)
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
42
PEGPH20 Enhanced Anti-PD-L1 Accumulation
in HA-high Ovarian Cancer Animal Model
SKOV3/HAS2 ovarian tumor model with anti-human-PD-L1-AlexaFluor 488
Anti-PD-L1
PEGPH20 +
Anti-PD-L1
p = 0.006
( B a s e lin e s u b t r a c te d )
M e a n d e n s it y ( S E M )
Individual Tumors
20000
15000
10000
5000
P
E
G
P
H
2
0
3
7
S
a
.5
li
µ
n
p
e
k
0
Anti-PD-L1-AlexaFluor 488 Fluorescence
Rosengren et al. (2016). AACR Annual Meeting, Poster #4886
43
PEGPH20 Immuno-Oncology Program Highlights
HA-high tumor status may lead to a more immunosuppressive TME
In HA-high syngeneic mouse tumors, PEGPH20 enhanced the effect
of immune checkpoint inhibitors
In HA-low mouse tumors, a combinatorial effect was not observed
PEGPH20 treatment increased anti-PDL1 antibody accumulation in
ovarian tumor xenograft model with elevated HA
44
Agenda
Time
Topic
5 minutes
Opening Remarks
10
minutes
Focus on the Tumor Microenvironment
10
minutes
10
minutes
PEGPH20 Immuno-Oncology
PEG-ADA2: PEGylated Adenosine Deaminase 2
10
minutes
HTI-1511: Anti-EGFR Antibody-Drug Conjugate
15
minutes
Questions and Answers
Presenter
Dr. Helen Torley
President and CEO
Dr. Michael LaBarre
VP, Chief Scientific Officer
Dr. Sanna Rosengren
Director, Immunology and
Cell Biology
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
All
45
American Association for Cancer Research
Investor Meeting
PEG-ADA2: PEGylated
Adenosine Deaminase 2
Dr. Christopher Thanos
Senior Director, Biotherapeutics Discovery
April 18, 2016
The Tumor Microenvironment
Cancer cell
Vasculature
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
47
The Tumor Microenvironment - Adenosine
Cancer cell
Vasculature
HA
Adenosine Generation
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
48
The Adenosine Pathway:
A Recognized Immune
Checkpoint Interaction
Pardoll DM, Nature
Reviews Cancer 12,
252-264 (April 2012)
49
Adenosine: A Key Suppressor of
Immune Cells in Tumor Microenvironment
ATP
Tumor
>100x Increase
Tumor Microenvironment Adenosine
~0.1 µM → ~50 µM
Adapted from Stagg & Smyth, Oncogene, 2010
50
Adenosine: A Key Suppressor of
Immune Cells in Tumor Microenvironment
ATP
ATP
Tumor
>100x Increase
Tumor Microenvironment Adenosine
~0.1 µM → ~50 µM
Adapted from Stagg & Smyth, Oncogene, 2010
51
Adenosine: A Key Suppressor of
Immune Cells in Tumor Microenvironment
ATP
CD39
ATP
Tumor
>100x Increase
Tumor Microenvironment Adenosine
~0.1 µM → ~50 µM
Adapted from Stagg & Smyth, Oncogene, 2010
52
Adenosine: A Key Suppressor of
Immune Cells in Tumor Microenvironment
AMP
ATP
CD39
ATP
Tumor
>100x Increase
Tumor Microenvironment Adenosine
~0.1 µM → ~50 µM
Adapted from Stagg & Smyth, Oncogene, 2010
53
Adenosine: A Key Suppressor of
Immune Cells in Tumor Microenvironment
AMP
ATP
CD39
CD73
ATP
Tumor
>100x Increase
Tumor Microenvironment Adenosine
~0.1 µM → ~50 µM
Adapted from Stagg & Smyth, Oncogene, 2010
54
Adenosine: A Key Suppressor of
Immune Cells in Tumor Microenvironment
AMP
ATP
CD39
CD73
Adenosine
ATP
Tumor
>100x Increase
Tumor Microenvironment Adenosine
~0.1 µM → ~50 µM
Adapted from Stagg & Smyth, Oncogene, 2010
55
Adenosine: A Key Suppressor of
Immune Cells in Tumor Microenvironment
B cells
dendritic cells
(-) BCR-induced NF-κB
activation
↑ IL-6, IL-8, TGFβ, VEGF, IDO
(-) IL-12
T cells
(-) TCR-induced NF-κβ, IL-2, IL-4, IFN-γ
(-) cytotoxicity
(+) anergy
(+) CD4+ T cell diff. to Foxp3+ or Lag-3+ Tregs
neutrophils
NKT cells
blocks adhesion / extravasation
(-) phagocytosis
(-) superoxide & nitric oxide
(-) IFN-γ
(+) IL-4, IL-10, TGF-β
AMP
ATP
mast cells
CD39
CD73
Adenosine
ATP
↑ IL-4, IL-8, IL-13
↑ VEGF
endothelial cells
(-) ICAM-1 & E-selectin
↑ VEGF and angiogenesis
↑ βFGF, IGF-1
Tumor
>100x Increase
Tumor Microenvironment Adenosine
~0.1 µM → ~50 µM
Saturation of Adenosine Receptor Checkpoints on Immune Cells
macrophages
(-) phagocytosis
(-) superoxide & nitric oxide
(-) TNF-α, IL-12, MIP-1α
↓ MHC class II
↑ IL-10, IL-6
NK cells
(-) IFN-γ
(-) cytotoxicity
Adapted from
Stagg & Smyth,
Oncogene, 2010
56
ADA2: An Approach to Deplete Adenosine in the TME
H20
NH4
ADA2
Catalyzes the deamination of adenosine to inosine1,2
Human, extracellular glycoprotein, ~120,000 MW1,2
Highly resistant to inactivation in plasma3
Produced in standard CHO cells3
1Zavialov
AV, Biochem J. 391, p51-7 (2005)
AV, J Biol Chem. 285(16):12367-77 (2010)
3Wang et al. (2016). AACR Annual Meeting, Poster #1217
2Zavialov
57
PEGylated ADA2: Improved Pharmacokinetics Profile
in Animal Model
ADA2-WT
200
Activity (U/ml)
PEG-ADA2-K374D
20
PEG-ADA2K374D extended PK profile in mice*
• t1/2 = 37.6 hours
• Less frequent dosing
• Systemic dosing
2
0
0
20
40
60
80
Time (Hour)
*Dosed at 3 mg/kg, n=9 mice/group
Wang et al. (2016). AACR Annual Meeting, Poster #1217
58
Tumor Growth Inhibition With PEG-ADA2 in
Colon Cancer Animal Model
V e h ic le
600
P E G A D A 2 -K 3 7 4 D
T u m o r V o lu m e , m m
3
(S E M )
Murine CT26 colon tumor model
400
p < 0.0001
200
0
0
2
4
6
D a y o n S tu d y
PEGADA2-K374D, 0.3 mg/kg
2X weekly, IV(N=8)
Wang et al. (2016). AACR Annual Meeting, Poster #1217
59
PEG-ADA2 Increased T-cell Infiltration in CT26 Colon
Cancer Animal Models
Murine CT26 colon tumor model
C D 3 p o s itiv e c e ll d e n s ity
p < 0.001
0 .4
5-fold increase
0 .3
0 .2
0 .1
0 .0
p re - d o se
6 - h o u rs p o st- d o se
Histological Assessment of T-cell infiltration
PEGADA2-K374D, 0.3 mg/kg
2X weekly, IV(N=8)
Wang et al. (2016). AACR Annual Meeting, Poster #1217
60
CD73 May Be a Useful Biomarker for Identifying
Tumor Types That Could Respond to PEG-ADA2
ATP
CD39
AMP
CD73
Adenosine
Wang et al. (2016). AACR Annual Meeting, Poster #1217
61
CD73 May Be a Useful Biomarker for Identifying
Tumor Types That Could Respond to PEG-ADA2
IM
T
4/
S
4+
o
M
H
19
is
ad
M
C
1/
n
P
-1
4T
09
5/
/S
O
C
IM
C
20
5/
N
L
K
N
L
E
M
T
20
-6
02
/S
S
C
/IM
C
an
02
an
P
C
/S
/IM
C
L
P
C
L
/IM
10
F
10
F
16
B
K
19
H
M
/S
IM
C
S
P
4+
n
o
is
ad
M
4/
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C
1/
/S
09
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4T
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L
K
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20
T
M
E
20
5/
/S
S
C
/IM
-6
02
an
an
P
P
02
/S
/IM
C
C
L
L
C
L
L
C
T
26
/S
/IM
/IM
C
10
/S
F
10
F
16
B
16
/S
0 .0 1
C
0 .0 1
L
0 .1
/IM
0 .1
L
1
26
1
T
10
C
10
T
100
IM
100
C
1000
B
Adenosine
CD73 Expression
in Various Tumors*
(
,
)
1000
v s C T 2 6 /H A S 3 IM
F o ld G e n e e x p r e s s io n
CD39 Expression in Various Tumors*
CD73
16
AMP
B
CD39
ATP
Various solid tumor types tested
*syngeneic mouse tumors
Wang et al. (2016). AACR Annual Meeting, Poster #1217
62
PEG-ADA2 Mediated Tumor Growth Inhibition in
CD73-Positive Animal Models
T u m o r V o lu m e , m m
(S E M )
KPC-derived pancreatic model
V e h ic le
1000
500
0
*
* p < 0.0001
0
5
10
D a y o n S tu d y
PEGADA2-K374D, 0.3 mg/kg
2X weekly, IV(N=8)
2000
V e h ic le
P E G A D A 2 -K 3 7 4 D
3
P E G A D A 2 -K 3 7 4 D
15
T u m o r V o lu m e , m m
1500
3
(S E M )
KLN-205 lung model
1500
*
1000
500
0
* p < 0.0001
0
5
10
15
20
D a y o n S tu d y
Wang et al. (2016). AACR Annual Meeting, Poster #1217
63
Engineering ADA2 Yielded a Variant With a
16-fold Improvement in Enzymatic Activity
S265
ADO
Analog
R222
PDB#
3LGG1
1Zavialov
2Wang
AV, J Biol Chem. 285(16):12367-77 (2010)
et al. (2016). AACR Annual Meeting, Poster #1217
64
Engineering ADA2 Yielded a Variant With a
16-fold Improvement in Enzymatic Activity
S265
ADO
Analog
R222
PDB#
3LGG1
kcat/KM (1/Ms)
Wild-Type
R222Q/S265N2
10,312
165,411
16x improved
1Zavialov
2Wang
AV, J Biol Chem. 285(16):12367-77 (2010)
et al. (2016). AACR Annual Meeting, Poster #1217
65
PEG-ADA2R222Q/S265N Inhibited Lung Metastasis
in Breast Cancer Animal Model
Mouse 4T1 breast cancer metastasis model
40
30
Lung
metastases
per mouse
20
10
Study day 18, histological assessment
n=6 mice (vehicle), n=8 mice (treatment)
2x weekly dosing, 0.3 mg/kg
Vehicle
1
0
p = 0.0183
PEG-ADA2
R222Q/S265N
1Wang
et al. (2016). AACR Annual Meeting, Poster #1217
66
Treatment With PEG-ADA2 Decreased TME Adenosine
Levels in Pancreatic Cancer Animal Model
KPC-derived pancreatic tumor model
30
TME
Adenosine
(µM)
20
10
p = 0.0127
0
U n tre a te d
P E G -A D A 2
T u m o rs
R 2 2 2 Q /S2 6 5 N
Measuring adenosine levels in a mouse TME (solid tumor)
KPC-derived mouse tumors
10 mm probe (55 kDa MWCO)
Microdialysis perfusates analyzed by LC-MS
1Wang
et al. (2016). AACR Annual Meeting, Poster #1217
from: Cancer Research 57, 2602-2605(1997)
2Protocol
67
PEG-ADA2 Program Highlights
Adenosine:
Attractive
immune
checkpoint
target
PEG-ADA2:
An engineered
human
enzyme that
targets
adenosine
• Abnormally high levels accumulate in the TME
• Binds to receptor checkpoints on immune cells
• Contributes to an immunosuppressive TME
• Improved pharmacokinetics and enzyme activity
• Anti-tumor responses observed in several animal
models
− Increase in T-cell infiltration
− Decrease in high TME adenosine levels
68
Agenda
Time
Topic
5 minutes
Opening Remarks
10
minutes
Focus on the Tumor Microenvironment
10
minutes
10
minutes
PEGPH20 Immuno-Oncology
PEG-ADA2: PEGylated Adenosine Deaminase 2
10
minutes
HTI-1511: Anti-EGFR Antibody-Drug Conjugate
15
minutes
Questions and Answers
Presenter
Dr. Helen Torley
President and CEO
Dr. Michael LaBarre
VP, Chief Scientific Officer
Dr. Sanna Rosengren
Director, Immunology and
Cell Biology
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
All
69
American Association for Cancer Research
Investor Meeting
HTI-1511: Anti-EGFR AntibodyDrug Conjugate
Dr. Christopher Thanos
Senior Director, Biotherapeutics Discovery
April 18, 2016
The Tumor Microenvironment
Cancer cell
Vasculature
HA
Collagen
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
71
The Tumor Microenvironment – Molecular Receptors
Cancer cell
Vasculature
HA
Collagen
Molecular Receptors
Macrophage
T cell
Treg
MDSC
Fibroblast
Adenosine
Molecular
Receptors
72
Two Limitations With Anti-EGFR Therapeutics
1
Treatment
can lead to
skin rash
• May limit dosing
2
Downstream,
activating
mutations
• KRAS mutations present in over 50% of mCRC4
• ~90% cutaneous side effects1-3
• BRAF mutations in ~10% of mCRC5
• EGFR mutations in ~3-19% of NSCLC in west6-7
1Cunningham,
NEJM 2004, 2Van Cutsem, JCO 2012, 3Amado, JCO 2008
Cancer Discovery 2014, 5Barras, Biomarkers in Cancer, 2015
6Deerden, Annals in Oncology, 7Lee, JNCI 2013
4Misale,
73
Physicochemical Properties Offer
Opportunities for TME-Specific Therapeutics
Tumor microenvironment
↓ pH vs. Healthy Tissue
↑ Lactate
↑ Albumin
Solid tumor example
Acidic pH
Blood vessel
(immunofluorescent)
Proliferation
(iododeoxyuridine)
Hypoxia
(pimonidazole)
1mm
Adapted from Lancet Oncol 2010; 11: 661–69
74
Halozyme mAb has Attenuated In Vitro Binding
to EGFR at Skin pH
EGFR Binding
(EC50 ng/mL)
50
40
30
Acidic pH, TME conditions
Skin pH
Low Affinity
HALO Mab
Cetuximab
20
10
0
High Affinity
pH 6.0
pH 6.5
pH 7.4
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
75
Halozyme mAb Attenuated Human Skin Binding
vs. Human Tumor Binding in Xenograft Models
Day 1
Day 2
Day 3
Cetuximab
Control
Comparable, strong
binding between tumor
and skin
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
76
Halozyme mAb Attenuated Human Skin Binding
vs. Human Tumor Binding in Xenograft Models
Day 1
Day 2
Day 3
Cetuximab
Control
Comparable, strong
binding between tumor
and skin
HALO
Anti-EGFR
mAb
Strong tumor,
attenuated skin binding
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
77
ADCs May Treat EGFR+ Mutation-Resistant Tumors
EGFR mediated
signaling promotes cell
growth
EGF
EGFR
KRAS
BRAF
MEK
ERK/MAPK
Proliferation
Migration
Angiogenesis
Survival
Adapted from Pao, Nature Reviews Cancer 10, 760-774 (2010)
78
ADCs May Treat EGFR+ Mutation-Resistant Tumors
EGFR mediated
signaling promotes cell
growth
Naked anti-EGFR mAb
inhibits signaling
pathway
Naked
mAb
EGF
EGFR
EGF
KRAS
KRAS
BRAF
BRAF
MEK
MEK
ERK/MAPK
ERK/MAPK
Proliferation
Migration
Angiogenesis
Survival
No signaling
Adapted from Pao, Nature Reviews Cancer 10, 760-774 (2010)
79
ADCs May Treat EGFR+ Mutation-Resistant Tumors
EGFR mediated
signaling promotes cell
growth
Naked anti-EGFR mAb
inhibits signaling
pathway
Mutation promotes cell
growth and is resistant to
mAb therapy
Naked
mAb
EGF
EGFR
EGF
KRAS
KRAS
KRASMut
BRAF
BRAF
BRAF
BRAF
MEK
MEK
MEK
ERK/MAPK
ERK/MAPK
ERK/MAPK
Proliferation
Migration
Angiogenesis
Survival
Proliferation
No signaling
Migration
Angiogenesis
Survival
Adapted from Pao, Nature Reviews Cancer 10, 760-774 (2010)
80
ADCs May Treat EGFR+ Mutation-Resistant Tumors
EGFR mediated
signaling promotes cell
growth
Naked anti-EGFR mAb
inhibits signaling
pathway
Mutation promotes cell
growth and is resistant to
mAb therapy
ADC overcomes
mutation resistance and
selectively kills tumor cell
ADC = antibody-drug conjugate
Naked
mAb
EGF
EGFR
ADC
EGF
KRASMut
KRAS
KRAS
KRASMut
BRAF
BRAF
BRAF
BRAF
MEK
MEK
MEK
MEK
ERK/MAPK
ERK/MAPK
ERK/MAPK
ERK/MAPK
Proliferation
Migration
Angiogenesis
Survival
Proliferation
No signaling
Migration
Angiogenesis
Survival
Internalized ADC
BRAF
Released Cytotoxins
Tumor Cell Death
Adapted from Pao, Nature Reviews Cancer 10, 760-774 (2010)
81
Tumor Regressions in KRAS- and BRAF-Mutated Tumor
Animal Models
Human TNBC Tumor Xenografts
MDA-MB-231M (KRASG13D)
Human CRC Tumor Xenografts
HT29 (BRAFV600E)
2500
2500
C e t u x im a b ( 3 0 m g /k g )
V e h ic l e
C e t u x im a b ( 3 0 m g /k g )
H A L O V a r ia n t 1 - M M A E ( 1 m g /k g )
H A L O V a r ia n t 1 - M M A E ( 1 m g /k g )
H A L O V a r ia n t 1 - M M A E ( 1 0 m g /k g )
H A L O V a r a n t 1 - M M A E ( 3 0 m g /k g )
1500
1000
500
H A L O V a r ia n t 1 - M M A E ( 3 m g /k g )
2000
H A L O V a r ia n t 1 - M M A E ( 1 0 m g /k g )
H A L O V a r ia n t 1 - M M A E ( 3 0 m g /k g )
3
H A L O V a r ia n t 1 - M M A E ( 3 m g /k g )
T u m o r V o lu m e (m m ) ± S E M
2000
3
T u m o r V o lu m e (m m ) ± S E M
V e h ic l e
1500
1000
500
6/6 Regressions, no evidence of tumors
0
0
10
20
30
↓
40
60
70
80
90
100
0
10
20
30
40
50
60
70
80
90
100 110
T im e ( D a y s )
T im e ( D a y s )
n=6 mice group
2X weekly dosing
↓
0
50
*Dosing stopped at Day 38
*Dosing stopped at Day 39
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
82
Next Generation ADC Technology
1st Generation Chemistries
# drugs / mAb
0
1
2
3 4 5
6
7 8 9
Random Lysine Conjugation
ThioBridge™
Cysteine Conjugation
# drugs / mAb
0
1
2
3 4 5
6
7 8 9
Cysteine Conjugation
Heterogeneous
# drugs / mAb
0
1
2
3 4 5
6
7 8 9
More homogeneous
Thanos, 2016 in press
83
HTI-1511: Anti-EGFR mAb Conjugated With
Thiobridge-MMAE Highly Homogeneous
mAU
4 Drug Conjugates per mAb (98.2%)
25.0
20.0
Analytical Hydrophobic
Interaction Chromatography
HALO
anti-EGFR
mAb
Chromatography Peak = HTI-1511
• HALO anti-EGFR mAb
• Thiobridge MMAE Chemistry
• Drug: Antibody Ratio = 4
15.0
10.0
4 Thiobridge
MMAE’s
5.0
12.4 min (1.8 %)
Retention time (minutes)
-2.0
2.5
5.0
7.5
10.0
12.5
15.0
17.5
20.0
22.5
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
84
Improved ADC Stability Observed in Primate Model
ADC Detection
mAb Detection
Toxin on ADC Detection
Control HALO mAb
1st Gen Conjugation
HTI-1511
400000
Concentration (ng/mL, Mean±SEM)
Concentration (ng/mL, Mean±SEM)
400000
300000
200000
100000
300000
200000
100000
0
0
0
10
20
30
Time (hours)
200 400 600
91% Intact ADC
Exposure Ratio (ER) =
Group Mean AUC of ADC / Group Mean AUC of Total x 100
0
10
20
30
Time (hours)
200 400 600
40% Intact ADC
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
85
Pilot Toxicology:
Safety Profile Tested in Primate Model
1 Cycle, 2 Dose Study Design, N=3 animals per group
Cycle 1
Week
Day
0
1
1
8
2
15
Endpoint
3
22
4
29
Dose
Parameters
•
•
•
•
•
•
•
•
Clinical observations and food consumption
Body weight
Dermal scoring
Clinical pathology
ECG and blood pressure
Veterinary physical examinations and ophthalmology
Histology
Pharmacokinetics
Limited dermal scoring findings comparable with vehicle control group
No unexpected findings observed at either dose (2.5 mg/kg and 8 mg/kg)
Safety profile met criteria for candidate nomination and further investment
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
86
Complete Tumor Regressions Observed in
Patient Derived (PDx) Tumor Models in Mice
2500
Cholangiocarcinoma (EGFR+, KRASpG12A) PDx
T u m o r V o lu m e ( m m ) ± S E M
3
V e h ic le
H T I-1 5 1 1
2000
3
T u m o r V o lu m e ( m m ) ± S E M
NSCLC (EGFR+, KRASpG12C) PDx
1500
1000
last dose
500
0
0
20
40
S tu d y D a y
N=8 mice / group
2.5 mg/kg (weekly dosing)
60
80
2500
V e h ic le
H T I-1 5 1 1
2000
1500
1000
500
last dose
0
0
20
40
60
S tu d y D a y
Huang,L. et al. (2016). AACR Annual Meeting, Poster #1472
87
HTI-1511 Anti-EGFR ADC Program Highlights
Engineered mAb with attenuated binding to human skin grafts
ADC mechanism targets KRAS- or BRAF-mutated tumors in mice
Utilization of next generation, Thiobridge chemistry
• More homogeneous, stable
Safety profile met criteria for candidate nomination
Complete tumor responses observed in PDx tumor models
IND enabling studies underway
88
Agenda
Time
Topic
5 minutes
Opening Remarks
10
minutes
Focus on the Tumor Microenvironment
10
minutes
10
minutes
PEGPH20 Immuno-Oncology
PEG-ADA2: PEGylated Adenosine Deaminase 2
10
minutes
HTI-1511: Anti-EGFR Antibody-Drug Conjugate
15
minutes
Questions and Answers
Presenter
Dr. Helen Torley
President and CEO
Dr. Michael LaBarre
VP, Chief Scientific Officer
Dr. Sanna Rosengren
Director, Immunology and
Cell Biology
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
Dr. Christopher Thanos
Senior Director,
Biotherapeutics Discovery
All
89
American Association for Cancer Research
Investor Meeting
Q&A
90