identification and prioritization of drug combinations for treatment of cancer
TRANSCRIPT
HORIZON DISCOVERY
Identification and Prioritization of Drug Combinations for Treatment of Cancer
Richard Rickles, Senior Director of Oncology
2
Presentation Overview
• Why are combination drugs important for treatment of cancer?
• Overview of cHTS screening strategy
• Example of cHTS screening results Amgen MDM2 inhibitor combination activities
• Combination drug leads- prioritization Ex vivo assays Tumor microenvironment assays Xenografts
• cHTS to identify synergies and antagonism
• Immuno-oncology
• Summary
3
Cancer Genome Efforts - the Complexity of Cancer
188 lung adenocarcinomas• 623 genes sequenced, 26 mutated at high frequency• Large number of chromosomal amplifications and deletions
Key pathways are mutated
TCGASomatic mutations affect key pathways in lung adenocarcinomaNATURE| Vol 455|23 October 2008
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Brain Cancer: Frequent Genetic Alterations in Critical Signaling
NATURE| Vol 455|23 October 2008TCGA
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The Complexity of Cell Signaling• Hard to predict tumor cell responses to cancer drugs
• Horizon Discovery’s strategies are robust can be an agnostic approach to probe cell signaling
The color of a node signifies the phenotypic effect of removing the corresponding protein (red, lethal; green, non-lethal; orange, slow growth; yellow, unknown).
Source: ibis.northwestern.edu/faculty/amaral.html
RealityCartoonSource: mayoclinicproceedings.com
ErbB1ErbB2ErbB3ErbB4PDGFR
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The Dose Matrix: Identification of Combination Activities
• Combination effects can manifest as potency shifts or efficacy boosts
• Challenging to pick concentrations and optimal ratios
• Dose-response matrix screening can detect both synergy and antagonism
Effec
t
Concentration
Measured Activities Null Interaction Model Excess Cytostatic
Cytotoxic
Inclusion of T0 measurement to help distinguish cytostatic versus cytotoxic activities
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Horizon Has an Extensive Cell Line Collection Available for Screening
>800 Core Cell Line Collection
• Core cell line collection has extensive overlap with the Cancer Cell Line Encyclopedia (CCLE) collection
• Wealth of information accumulated about use of cell line collection for HTS
• Augmented by X-MAN® isogenic cell lines
Cell Line Collection
>550 X-MAN® Isogenic Cell Line Pairs+
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Horizon has Worked with Many Leading Pharma Companies and Research Foundations
• Many examples of multiple projects over a multi-year time span
• Quick delivery time• Quality of data• Cost-effective
• CRO as a collaborator• Expertise• Presentations at meeting• Publications• Assist with IP filings
CHDI (HD)SMACystic FibrosisDMD
Examples ofDiscovery Collaborations
Systematic identification of synergistic multi-target mechanisms conferring selectivity and
enabling patient stratification
Drug leads
Examples ofDiscovery/Translation Collaborations
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Custom Offerings
Pairwise Screening
• 100-200 compounds of diverse mechanism and activity
• >20,000 pairwise combinations• 50-100s of cell lines
Enhancer Screening
• One or more focus compounds• 100-250 enhancer compounds of
diverse activity and mechanisms• 20-100 cell lines
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Alternative Screening Patterns Are Available• Our automated platform allows screening formats to be
adapted to best suit the experimental design
• Non-uniform sampling of combination space Increases cHTS efficiency Decreases cHTS-associated costs Decreases timeline
Co
mp
ou
nd
A
Compound B
6x6 Optimized
Co
mp
ou
nd
A
Compound B
6x6 Format
Co
mp
ou
nd
A
Compound B
HFDR Format
Patch 5 4x4Diag2x2_ic20 2x62x2top
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Comparison of Full versus HFDR Dose Matrix
Validation- alternative combination dose matrices behave similarly to full dose matrices when compared experimentally
– HFDR highly correlates with 6x6 format (mean R=0.96)
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cHTS Case Study: Amgen and MDM2 (Discovery Effort)
• MDM2 inhibitors hold great promise as cancer therapeutics
• As with many other targets, drug resistance will likely limit efficacy as single agents
• Amgen engaged Horizon to systematically identify and verify multi-target synergies
Phase 1: Primary ScreenMDM2iX
1200 Enhancers(SOC, Emerging Rx, Probes)
X 10 Cell Lines
> 25,000Replicate matrixes
Analyzed by Chalice™
Synergy ScoreHeat map showing synergy score
Identified multiple synergistic interactions
2014 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4039142/
MDM2 antagonists synergize broadly and robustly with compounds targeting fundamental oncogenic signaling pathways Oncotarget 2014
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Case Study Amgen MDM2: Identify, Verify, and Determine Synergy
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Analysis of Combination Activities Using Secondary Assays
Inhibition (%)
CRx5
01 (
uM)
0.0
012
5e-3
.02
.08
Dexamethasone (uM)0 .062 .25 1 4
N=
1
0
-7.4
-3.9
10
6.3
13
25
37
33
37
48
60
66
72
79
83
36
45
54
65
70
77
82
87
37
44
48
65
70
76
84
86
36
41
52
61
72
79
82
85
37
44
47
63
68
77
82
84
37
41
47
62
73
74
83
87
42
51
58
66
73
80
83
86
ADD Excess Inhibition (%)CRx5
01 (
uM)
0.0
012
5e-3
.02
.08
Dexamethasone (uM)0 .062 .25 1 4
Vol
=14
.5(.
07)
C
hi2=
1100
0
-7.5
-4.4
8.7
.5
-1.9
-2
2.7
1.3
5.6
16
27
33
38
43
46
-1.3
8.1
18
28
33
40
44
49
-1.1
5.2
10
26
32
38
46
47
-2.5
2.3
13
22
33
40
44
46
-1.5
5.2
8.2
24
29
38
43
45
-1.5
1.9
8.2
23
34
36
44
49
3.4
12
19
28
34
42
44
47
Tumor cells from a patient resistant to Velcade, Revlimid, dexamethasone and transplant
Ex Vivo Assay
CGS-
2168
0 uM
CGS-
2168
0 uM
Survival
0 25 50 75 1000
25
50
75
100
p=0.0002
Days after treatment
Per
cent
Sur
viva
l
Vehicle
CRx-501
Dex
Dex + CRx-501
Xenograft assay
Enhanced Survival
Microenvironment Assays No IL-6
Rickles, et.al. Blood (2010)Rickles, et.al. MCT (2012)
+ IL-6
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Flexible System Able to Detect Both Synergies and Antagonism
SYNERGY
ANTAGONISM
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The Past and the Future of Cancer Drug Development
Development of chemotherapeutic drugs better efficacy with use of drug combinations• Pair drugs that exhibit single agent activity in the clinic• Pair drugs based on knowledge of disease biology
Development of targeted drugs better efficacy with use of drug combinations• Pair drugs that exhibit single agent activity in the clinic• Pair drugs based on knowledge of disease biology• Pair drugs based on combination drug activities (cHTS, preclinical models)
Development of immuno-modulatory drugs better efficacy with use of drug combinations
• Pair drugs that exhibit single agent activity in the clinic• Pair drugs based on knowledge of disease biology• Pair drugs based on combination drug activities (cHTS, preclinical models)
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cHTS to Identify Immune Modulators
• Horizon Discovery has extensive experience with inflammation assays
• Example: identification of synergistic drug combinations that inhibit production of pro-inflammatory cytokines
Measured Activities Excess (TNFa)
0
20
40
60
80
100
0
20
40
60
80
100
% In
hibi
tion
% In
hibi
tion
TCR activation(anti-CD28/
anti-CD3 stimulation)
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Combination Drug Discovery
Assays need to be robust • Known immuno-modulatory agents should exhibit activity
Capture disease complexity • If not primary screen, with use of secondary screen(s)
Combination drug discovery is “critical path” for cancer drug development
• Synergistic selective killing of tumor cells• Immuno-modulatory drugs
Horizon Discovery• More than a decade of experience with oncology and
inflammation cHTS• Strong TIDVAL capabilities (shRNA, siRNA, sgRNA)• Strong track record for technology development• Proven ability to quickly deliver actionable insights• insights
Your Horizon Contact:
t + 44 (0)1223 655580f + 44 (0)1223 655581e [email protected] www.horizondiscovery.comHorizon Discovery, 7100 Cambridge Research Park, Waterbeach, Cambridge, CB25 9TL, United Kingdom
Richard RicklesSenior Director, [email protected]+1 (0)