Computational Chemistry Mechanistic Enzymology
Biomarker Development
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Cellular Biology & Pathway Interrogation Assay Development & Validation
Medicinal Chemistry & Synthesis
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R2 = 75% (canine)DOGRAT
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Benefits of Contracting CDT
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
Research Partnership from Target ID through IND and beyond
Computational & Medicinal Chemistry
Experience where it counts:– Drug Design– Synthetic Route Analysis– Compound Synthesis– Structure-Based Drug Design – Molecular Modeling– Virtual Screening
Confluence can enable at all stages:– 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
Lys 53
Asp 168
Glu 71
Met 109
Ala 51
Thr 106
Asp 112 Ser 154
Leu 167
Lys 53
Asp 168
Glu 71
Met 109
Ala 51
Thr 106
Asp 112 Ser 154
Leu 167
Assay Establishment and ValidationWe 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
Biochemistry/Enzymology
Mechanism of Action Studies
Enzyme Activity
Steady State Kinetics
Inhibitory Mechanism
CompetitiveNoncompetitiveUncompetitive
Irreversible (kinact & Ki)Slow tight binding (kon & koff)
Aggregate (non drugable)
Thermodynamics
Isothermal Titration Calorimetry
Independent of Catalysis
Kd and stoichiometry DH, DS, DG
Direct Binding
RadioligandBiacore
TR-FRETFluorescence (FP)
AbsorbanceLuminescence
Filtration
Independent of Catalysis
Binding kinetics (kon & koff)
Key to Increased Efficiency of Lead Discovery & Development is to Link SAR to Mechanism of Action!
Cell-Based Assay Design & Development
Cell-based assays are invaluable for drug discovery:– Identification of biologically important signal transduction
pathways and key point(s) for therapeutic intervention– Understanding compound mechanism of action– Quantitation of efficacy and selectivity– Understanding safety issues linked to pathways
Cell-based assays have inherent problems in:– Simulating ex vivo the microenvironment/disease
state/species and/or tissue– Choosing disease relevant cell types and/or stimuli– Genetic drift, inter-donor variability, cross-species, etc.– Understanding effects of enzyme isoforms, scaffolding,
substrate selectivity, etc.
Confluence scientists have experience with these issues:– We can identify & design validated assays for translatable
target modulation & efficacy biomarkers– Offer broad assay technologies including DELFIA/FRET,
AlphaScreen, Westerns, FACS etc.– Biomarkers selected for assay development based on
pertinent scientific literature and our collective experience
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Intracellular and Secreted Analyte Analysis
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Coupling Assay Technology with Biological Relevance
Biomarker Development
CDT is Experienced With:– Target, Mechanism and Disease Biomarkers– Biomarker selection– Biomarker assay design, development,
validation and interpretation– Cell surface, intracellular and secreted analytes
CDT Can Deliver:– Assays developed across species and in
various biological matrices– Complete statistical evaluation of data– Seamless assay transfer to the clinical CRO
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.
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We can help you generate an interpretable and decision-enabling cellular, animal or human study though effective use of translatable biomarkers.
Biology Facilities and Instrumentation
Facilities
• General Biology Lab• Cell Culture Lab• WUSTL Screening
Core• Radiation Laboratory
Analytical
• Biacore 2000• Spectramax Plus• Spectramax M2• Analyst HT 96/384• Caliper LabChip• Packard Top Count• Tecan GENios• Microcal ITC• BMG Polar Star Omega• Waters & Agilent HPLC• FACScan Cytometer• MesoScale Discovery• Luminex
Other
• SDS PAGE/Western• Cell culture hoods• CO2 Incubators
• Various Microscopes• Centrifuges of All
Types• Various Balances• Market Forge
Autoclave• Thermocyclers• Shaking Incubators• Static Incubators• Biological Safety
Hoods
Automation
• Biomek NX:96/384• Precision 2000• Multidrop Combi
Chemistry Facilities & Instrumentation
Facilities & Infrastructure
• Synthetic Chemistry Lab• Analytical/Purification
Lab• Washington University facilities agreement• Waste and storage
facilities for chemicals
Synthesis Purification & Analytical• Air Sensitive Reactions• Customized Quantities (mg-g)• Library
Design/Production• Intermediate
Preparation• Route Identification• Route Optimization
• HPLC• Normal Phase
Purification• Reverse Phase Purification• LC/MS• NMR
Management: Joseph Monahan, PhD, President Michael Kuo, MD, Board Member Al Beardsley, PhD, Business Development Walter Smith, MS, Vice President Research
Scientist Founders: Gabriel Mbalaviele, PhD Shaun Selness, PhD
Business Professionals: Jennifer Bohnert – Controller Lisa Gabel - Bookkeeper Polsinelli Shughart - General Counsel Harness, Dickey & Pierce - IP Attorney Stone Carlie - Tax Advisor
Confluence Team
Staff Scientists: Barry Burnette, PhD Jeff Hirsch, BS Balekudru Devadas, PhD Susan Hockerman, MS John Springer, MS Jerry Cubbage, PhD Jim Blinn, MS Heidi Hope, PhD Arthur Wittwer, PhD William Hood, MS Sheri Bonar, BA Matt Saabye, BS Jon Jacobsen, PhD Taryn Syrtees, BS Kristin Sauer, BS
Joseph Monahan, PhDPresidentConfluence Discovery TechnologiesCORTEX Bioscience District4320 Forest Park Avenue, Suite 303St. Louis MO 63108josephmonahan@confluencelifesciences.comwww.confluencediscovery.com314.932-4032 x-305
Walter SmithVice President ResearchConfluence Discovery TechnologiesCORTEX Bioscience District4320 Forest Park Avenue, Suite 303St. Louis MO 63108waltersmith@confluencelifesciences.comwww.confluencediscovery.com314.932-4032 x-305