Capabilities presentation for Confluence Discovery Technologies

1. Computational Chemistry Medicinal Chemistry & Synthesis Mechanistic Enzymology
100.000
100
DOG
R2 = 75% (canine) 90 Target
Mechanism
% Control Response
RAT 80
10.000 MOUSE
70 Disease
Target IC50
60
1.000
50
40
0.100 30
20
10
0.010
0.1 1 10 100
Drug Dose (mg/kg)
0.001
0.01 0.10 1.00 10.00 100.00
Lymphocyte assay IC50
Cellular Biology & Pathway Interrogation Assay Development & Validation Biomarker Development

2. Benefits of Contracting CDT
Research Partnership from Target ID through IND and beyond
 Problem Solvers
– Experienced drug discovery team
– Broad Technology Base
 Mechanistic enzymology – industry leading
 Biochemical and cell biology assay development
 Biomarker assay development
 Computational and Medicinal Chemistry
 Partners
– Communication
– Versatile and flexible
– Project leadership/consultation – partner in strategy and execution

3. Computational & Medicinal Chemistry
 Experience where it counts: Ala 51
– Drug Design Thr 106
– Synthetic Route Analysis Lys 53
– Compound Synthesis Met 109
Glu 71
– Structure-Based Drug Design
– Molecular Modeling Leu 167
– Virtual Screening Asp 168
 Confluence can enable at all stages: Asp 112 Ser 154
– Understanding the Competitor Landscape
– Discovering Areas with Freedom To Operate
– ID Leads from Virtual Screen
– Parachute Opportunity Exploitation
– Designing Initial Chemical Matter
– Identifying Synthetic Routes
– Providing Active Pharmaceutical Ingredients
– Writing Patent Protocols
– Optimizing Current Lead Matter
– Singleton and Library Production

4. Assay Establishment and Validation
We can deliver a validated assay where no off the shelf option exists
 Assay Development Expertise
– Flexible screening formats
– Biochemical and Cellular assays
– Direct HTS translation
 Appropriately Equipped
– 96 and 384 well enabled
– Multiple liquid handling stations
– Versatile plate readers
 Data Analysis and Secondary Assays
– Hit validation and IC50 values
– Mechanistic characterization of hits
– Follow-up orthogonal assays
 Problem Solving
– New targets – need entire testing funnel
– Complex signaling systems
– Challenging targets
– Exclusion of artifacts
 Medium Throughput Specialty Screens

5. Biochemistry/Enzymology
Mechanism of Action Studies
Enzyme Activity Direct Binding Thermodynamics
Radioligand
Biacore Isothermal Titration
Steady State Kinetics
TR-FRET Calorimetry
Fluorescence (FP)
Absorbance
Luminescence
Inhibitory Mechanism Filtration
Competitive
Noncompetitive
Independent of Catalysis
Uncompetitive Kd and stoichiometry
Irreversible (kinact & Ki) Independent of Catalysis DH, DS, DG
Slow tight binding (kon & koff)
Binding kinetics (kon & koff)
Aggregate (non drugable)
Key to Increased Efficiency of Lead Discovery & Development is to Link SAR to Mechanism of Action!

6. Cell-Based Assay Design & Development
 Cell-based assays are invaluable for drug discovery: Intracellular and Secreted Analyte Analysis
– Identification of biologically important signal transduction 120 120
pathways and key point(s) for therapeutic intervention Protein Activity
100 100
% of Stimulated Control)
% of Stimulated Control)
Intracellular Analyte
– Understanding compound mechanism of action Secteted analyte,
80 80
– Quantitation of efficacy and selectivity
60 60
– Understanding safety issues linked to pathways
40 40
 Cell-based assays have inherent problems in:
20 20
– Simulating ex vivo the microenvironment/disease
state/species and/or tissue 0
- Stim 0mM 0.01mM 0.1mM 1.0mM 10mM
0
[Inhibitor]
– Choosing disease relevant cell types and/or stimuli
+Stimulation
– Genetic drift, inter-donor variability, cross-species, etc.
– Understanding effects of enzyme isoforms, scaffolding, Cellular Quantitation of Kinase Inhibitor Selectivity
substrate selectivity, etc. Kinase 1
200
Kinase 2
 Confluence scientists have experience with these issues:
Phospho/Total (% Control)
Kinase 3
150
– We can identify & design validated assays for translatable
target modulation & efficacy biomarkers
100
– Offer broad assay technologies including DELFIA/FRET,
AlphaScreen, Westerns, FACS etc. 50
– Biomarkers selected for assay development based on
pertinent scientific literature and our collective experience 0
Coupling Assay Technology with Biological Relevance Kinase Inhibitor (nM)

7. Biomarker Development
Reliable interpretation of preclinical disease and pharmacology models
and human clinical studies requires suitable biomarker analysis to
optimally relate PK/PD, PK/TD and outcomes.
 CDT is Experienced With: 100
– Target, Mechanism and Disease Biomarkers 90 Target
Mechanism
% Control Response
80
– Biomarker selection Disease
70
– Biomarker assay design, development, 60
validation and interpretation 50
– Cell surface, intracellular and secreted analytes 40
30
 CDT Can Deliver:
20
– Assays developed across species and in 10
various biological matrices
0.1 1 10 100
– Complete statistical evaluation of data
Dose (mg/kg)
– Seamless assay transfer to the clinical CRO
We can help you generate an interpretable and decision-enabling cellular,
animal or human study though effective use of translatable biomarkers.

8. Biology Facilities and Instrumentation
Facilities Other
• General Biology Lab • SDS PAGE/Western
• Cell Culture Lab • Cell culture hoods
• WUSTL Screening Core • CO2 Incubators
• Radiation Laboratory • Various Microscopes
• Centrifuges of All Types
Analytical • Various Balances
• Market Forge Autoclave
• Biacore 2000 • Thermocyclers
• Spectramax Plus • Shaking Incubators
• Spectramax M2 • Static Incubators
• Analyst HT 96/384 • Biological Safety Hoods
• Caliper LabChip
• Packard Top Count
• Tecan GENios
• Microcal ITC
• BMG Polar Star Omega Automation
• Waters & Agilent HPLC • Biomek NX:96/384
• FACScan Cytometer • Precision 2000
• MesoScale Discovery • Multidrop Combi
• Luminex

9. Chemistry Facilities & Instrumentation
Facilities & Infrastructure Synthesis Purification & Analytical
• Air Sensitive Reactions
• Synthetic Chemistry Lab • HPLC
• Customized Quantities
• Analytical/Purification Lab • Normal Phase Purification
(mg-g)
• Washington University • Reverse Phase
• Library Design/Production
facilities agreement Purification
• Intermediate Preparation
• Waste and storage • LC/MS
• Route Identification
facilities for chemicals • NMR
• Route Optimization

10. Confluence Team
Management:
Staff Scientists:
 Joseph Monahan, PhD, President
 Barry Burnette, PhD
 Michael Kuo, MD, Board Member
 Jeff Hirsch, BS
 Al Beardsley, PhD, Business Development
 Balekudru Devadas, PhD
 Walter Smith, MS, Vice President Research
 Susan Hockerman, MS
Scientist Founders:  John Springer, MS
 Gabriel Mbalaviele, PhD  Jerry Cubbage, PhD
 Shaun Selness, PhD  Jim Blinn, MS
 Heidi Hope, PhD
Business Professionals:  Arthur Wittwer, PhD
 Jennifer Bohnert – Controller  William Hood, MS
 Lisa Gabel - Bookkeeper  Sheri Bonar, BA
 Polsinelli Shughart - General Counsel  Matt Saabye, BS
 Harness, Dickey & Pierce - IP Attorney  Jon Jacobsen, PhD
 Stone Carlie - Tax Advisor  Taryn Syrtees, BS
 Kristin Sauer, BS

11. Joseph Monahan, PhD Walter Smith
President Vice President Research
Confluence Discovery Technologies Confluence Discovery Technologies
CORTEX Bioscience District CORTEX Bioscience District
4320 Forest Park Avenue, Suite 303 4320 Forest Park Avenue, Suite 303
St. Louis MO 63108 St. Louis MO 63108
josephmonahan@confluencelifesciences.com waltersmith@confluencelifesciences.com
www.confluencediscovery.com www.confluencediscovery.com
314.932-4032 x-305 314.932-4032 x-305