Answer four fundamental questions on how to develop the most innovative cancer immunotherapy treatments, starting with screening for lead molecules and ending with evaluation of combination therapies.
2. 2
The Cancer-Immunity Cycle & Targeted Therapies
Multiple potential intervention steps to develop anti-cancer therapies
Oncology Meets Immunology: The Cancer-Immunity Cycle. Immunity. 39 (1): 1-10
3. 3
Fundamental questions for drug development
Screening &
Lead
Optimization
Efficacy &
Biomarker
Selection
Safety &
Pre-clinical
Studies
Clinical
Combination
Strategies
How do I screen for a functionally active lead molecule?
How can I determine efficacy in a human tumor microenvironment (TME)?
Is my drug active outside the context of the TME?
How does my drug act in a combination therapy?
Translating Discoveries into Drugs
4. 4
Target-based & Phenotypic Assays from DiscoverX
Products and services to support drug development
0% KinasesGPCRs Interleukins Pathways Epigenetics Checkpoint
100%
Primary Cell Profiling
DepthofCoverage
284
410
40
35
34
6
5. 5
From drug discovery to clinical combinations
Screening &
Lead
Optimization
Efficacy &
Biomarker
Selection
Safety &
Pre-clinical
Studies
Clinical
Combination
Strategies
Enabling Anti-cancer Immunotherapies
PathHunter®
Checkpoint Assays
PathHunter® Bioassays for
QC Lot Release Testing
BioMAP®
Oncology Systems
BioMAP
Diversity PLUS™
BioMAP®
Combo ELECT
6. 6
Challenges:
Requires specific cell-based assays for each target
Requires human cells with complex TCR signaling
Donor variability
Longer assay times and complicated protocols
Screening &
Lead
Optimization
Efficacy &
Biomarker
Selection
Safety &
Pre-clinical
Studies
Clinical
Combination
Strategies
How Do I Screen for Functionally Active Lead Molecules?
7. 7
Cancer Cell
Immune Cell
EFC Technology
PD-1 Signaling Assay
PathHunter® Checkpoint Assays
Ideal for screening and lead optimization
8. 8
1 0 -1 0
1 0 -9
1 0 -8
1 0 -7
1 0 -6
1 0 -5
1 0 -4
0
2 0 0 0 0
4 0 0 0 0
6 0 0 0 0
8 0 0 0 0
A n ti-P D -1 D ru g [g /m L ]
RLU
P e m b ro lizu m ab
N iv o lu m ab
HillSlope
IC50
Pembrolizumab
-2.010
6.630e-009
Nivolumab
-1.921
9.398e-009
Robust Screening & Lead Optimization Platform
1 0 -1 1
1 0 -1 0
1 0 -9
1 0 -8
1 0 -7
1 0 -6
1 0 -5
1 0 -4
0
1 0 0 0 0 0
2 0 0 0 0 0
3 0 0 0 0 0
4 0 0 0 0 0
RLU
C o m p o u n d [M ]
X L -2 8 8 0
D a s a tin ib
HillSlope
IC50
Dasatinib
-1.979
5.762e-009
XL-2880
-1.045
4.740e-007
Screen Biologics in
the PathHunter PD-1 Assay
Screen Small Molecule Inhibitors in
the PathHunter PD-1 Assay
PathHunter® assays support both biologics and small molecules
9. 9
PD-1
OX40
CD40
HVEM
VISTA
TIM3
Biologically relevant responses for co-stimulatory & co-inhibitory agents
Easy protocol with results in <5hrs
Supports development of biologics and small molecules
Screen confidently with highly sensitive response
Applicable for QC lot release of biologic drug
Summary for PathHunter® Checkpoint Assays
1 0 -1 2
1 0 -1 1
1 0 -1 0
1 0 -9
1 0 -8
1 0 -7
1 0 -6
1 0 -5
0
5 0 0 0 0
1 0 0 0 0 0
1 5 0 0 0 0
O X 4 0 S ig n a lin g A s s a y
O X 4 0 L [g /m L ]
RLU
+ O X 4 0
n o O X 4 0
Assays currently available
Enabling screening & lead optimization for co-stimulatory & co-inhibitory molecules
10. 10
Challenges:
In vitro assays do not mirror the complexity of TME
Animal-xenograft models are not high throughput
Lack of appropriate response biomarkers
How Can I Determine Efficacy in a Human TME?
Screening &
Lead
Optimization
Efficacy &
Biomarker
Selection
Safety &
Pre-clinical
Studies
Clinical
Combination
Strategies
11. 11
Large or Small
Molecules and
Combinations
BioMAP® Models of Human Disease Biology
Validated systems that provide predictive results
BioMAP Data &
Knowledgebase
56+ BioMAP
co-culture systems
Human Primary Cells
12. 12
Development of BioMAP® Human TME Models
BioMAP systems mirror intratumoral immune suppression
48h Assay Incubation
18. 18
Both Large And Small Molecules Can Be Tested
Screen and prioritize drug candidates
Anti-CD73 antibodies and Paclitaxel show increased immune cytokine production
CD87/uPAR
CEACAM5/CD66e
CollagenI
CollagenIII
Keratin20
P
PBMCCytotoxicity
sGranzymeB
sIFNg
SRB
sVEGF
tPA
uPA
CD40
CD87/uPAR
CEACAM5/CD66e
CollagenIV
Keratin20
PBMCCytotoxicity
sGranzymeB
sIFNg
SRB
Profiles
Paclitaxel, 14 nM
,
StroHT29 VascHT29
sIFNg
sTNFa
sVEGF
uP
sIFNg
Profiles
Paclitaxel, 14 nM
,
sIFNg
sTNFa
sVEGF
uP
sTNFa
sIFNg
StroHT29 VascHT29
Anti-Human CD73 was profiled as part of a collaborative study with MedImmune/AZ
Protein Biomarker Readouts
RelativeExpressionLevels
|Log10ratio|drug/vehiclecontrol)
19. 19
Challenges:
Current approaches rely on single cell or organ types
Bias towards specific targets and target pathways
Less complex and often isolated biology
Limited testing of broad-scale human biology with drug
Is My Drug Active Outside the Context of the TME?
Screening &
Lead
Optimization
Efficacy &
Biomarker
Selection
Safety &
Pre-clinical
Studies
Clinical
Combination
Strategies
20. 20
F
P
f
P
F
f
f
K
P
tP
P
P
C
K
P
tP
P
F
P
P
f
C
P
f
P
P
P
LogRatio
Profiles
Erlotinib, 1.1 uM
Erlotinib, 370 nM
3C 4H LPS SAg BT BF4T BE3C CASM3C HDF3CGF KF3CT MyoF lMphg
uPAR uPAR
P
tPA
uPA
uPAR
EGFR
F
P
f
P
F
f
f
K
P
tP
P
P
C
K
P
tP
P
F
P
P
f
C
P
f
P
P
P
LogRatio
Profiles
Erlotinib, 1.1 uM
Erlotinib, 370 nM
3C 4H LPS SAg BT BF4T BE3C CASM3C HDF3CGF KF3CT MyoF lMphg
uPAR
Eot3
uPAR
IL1a
MMP1
MMP9
P
tPA
uPA
uPAR
EGFR
IL8
Evaluation of Erlotinib Effects Outside the TME
Biomarker activity of EGFR-inhibitors
• Inform mechanism of action
• Identify biomarkers of efficacy
Activities detected at both concentrations are annotated
RelativeExpressionLevels
|Log10ratio|drug/vehiclecontrol)
Monocyte
Activation
T cell
Activation
B cell
Activation
Macrophage
Activation
Matrix-modulation, fibrosis, tissue remodeling
responses
Vascular EC
Inflammation
Epithelial Inflammation and Matrix
Remodeling
Vascular SM
Inflammation
21. 21
F
P
f
P
F
f
f
K
P
tP
P
P
C
K
P
tP
P
F
P
P
f
C
P
f
P
P
P
LogRatio
Profiles
Paclitaxel, 41 nM
Paclitaxel, 4.6 nM
3C 4H LPS SAg BT BF4T BE3C CASM3C HDF3CGF KF3CT MyoF lMphg
TF uPAR
uPAR
sIgG
F
P
f
P
F
f
f
K
P
tP
P
P
C
K
P
tP
P
F
P
P
f
C
P
f
P
P
P
LogRatio
Profiles
Paclitaxel, 41 nM
Paclitaxel, 4.6 nM
3C 4H LPS SAg BT BF4T BE3C CASM3C HDF3CGF KF3CT MyoF lMphg
TF uPAR
IL8
uPAR
VEGFR2
sIgG
CD69
Proliferation
Proliferation
Proliferation
Evaluation of Paclitaxel Outside the TME
Biomarkers for cardiovascular AE, anti-angiogenic and anti-metastatic potential
• Inform mechanism of action
• Identify activities that may highlight potential for adverse events
Activities detected at both concentrations are annotated
RelativeExpressionLevels
|Log10ratio|drug/vehiclecontrol)
Monocyte
Activation
T cell
Activation
B cell
Activation
Macrophage
Activation
Matrix-modulation, fibrosis, tissue remodeling
responses
Vascular EC
Inflammation
Epithelial Inflammation and Matrix
Remodeling
Vascular SM
Inflammation
22. 22
F
P
f
P
F
f
f
K
P
tP
P
P
C
K
P
tP
P
F
P
P
f
C
P
f
P
P
P
LogRatio
Profiles
Pembrolizumab, 2000 ng/ml
Pembrolizumab, 400 ng/ml
3C 4H LPS SAg BT BF4T BE3C CASM3C HDF3CGF KF3CT MyoF lMphg
F
P
f
P
F
f
f
K
P
tP
P
P
C
K
P
tP
P
F
P
P
f
C
P
f
P
P
P
LogRatio
Profiles
Pembrolizumab, 2000 ng/ml
Pembrolizumab, 400 ng/ml
3C 4H LPS SAg BT BF4T BE3C CASM3C HDF3CGF KF3CT MyoF lMphg
Evaluation of Pembrolizumab Outside the TME
Anti-PD1 antibodies are not inherently inflammatory
• Confirm selectivity
• Determine on/off target effects and even target related secondary effects
• Inform mechanism of action
• Nivolumab has an identical profile in this panel
Monocyte
Activation
T cell
Activation
B cell
Activation
Macrophage
Activation
Matrix-modulation, fibrosis, tissue remodeling
responses
Vascular EC
Inflammation
Epithelial Inflammation and Matrix
Remodeling
Vascular SM
Inflammation
No activities detected
RelativeExpressionLevels
|Log10ratio|drug/vehiclecontrol)
23. 23
Challenges:
Pre-clinical testing is technically challenging
Current models are poorly predictive & expensive
Often directly tested in patients
Screening &
Lead
Optimization
Efficacy &
Biomarker
Selection
Safety &
Pre-clinical
Studies
Clinical
Combination
Strategies
How Do My Drugs Act in Combination Therapy?
24. 24
Combination Matrix
Enhanced Immune Response with Combinations
RelativeExpressionLevels
|Log10ratio|drug/vehiclecontrol)
Protein Biomarker Readouts
Pembrolizumab plus Paclitaxel combination enhances cytokine production
25. 25
Lead Molecule:
PathHunter® assays can help identify and optimize a lead candidate
Efficacy:
BioMAP® Oncology Systems determine drug efficacy in a human TME model
Safety:
Diversity PLUS™ evaluates drug activity outside the context of the TME
Combinations:
Combo ELECT tests the impact of drug interactions in a human TME model
Summary
Screening &
Lead
Optimization
Efficacy &
Biomarker
Selection
Safety &
Pre-clinical
Studies
Clinical
Combination
Strategies
26. 26
DiscoverX: Enabling Testing to Therapy
www.discoverx.com
Send your questions to :
SupportUS@discoverx.com
SupportEurope@discoverx.com