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© NCMM - Centre for Molecular Medicine Norway www.ncmm.uio.no

EATRIS – European Advanced

Translational Research

Infrastructure in Medicine

www.ncmm.uio.no 2

www.ncmm.uio.no 3 www.ncmm.uio.no 4

www.ncmm.uio.no 5 www.ncmm.uio.no 6

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www.ncmm.uio.no 7 www.ncmm.uio.no 8

www.ncmm.uio.no 9 www.ncmm.uio.no 10

www.ncmm.uio.no 11 www.ncmm.uio.no

EATRIS i Norge

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Nasjonal styringsgruppe

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www.ncmm.uio.no 13

European Infrastructure of Open

Screening Platforms for Chemical Biology

www.eu-openscreen.eu

EU-OPENSCREEN – Executive Summary February 2013

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Chemical Biology

X © HZI/F. Sasse

Investigation of biological systems using

chemical tools

MISSION

EU-OPENSCREEN’s objective is the development of novel research

‘tool’ compounds for all fields of the Life Sciences.

Tool compounds enable researchers to investigate molecular mechanisms of physiological

and pathological processes, many of which can only be studied with these chemical ‘tools’.

Chemical tools complement methods of molecular biology, such as mutagenesis or RNA

interference.

All generated tools and data will be made publically available to the scientific community.

EU-OPENSCREEN – Executive Summary February 2013 Page 16

Chemical compounds, such as this

small molecule, can bind to cellular

structures (e.g. proteins) and modulate

their functions.

Chemical keys for life’s locks

(Application in

cancer treatment)

Example:

STRATEGY

Chemistry Compounds

Biology Assays & Targets meets

Partner Sites

Screening Technology

Compound Profiling

Chemistry Services

Compound Management

EU-OPENSCREEN-ERIC

Compound Collection

Project Management

Database

Training & Education

User User

Activity

profiles

Tool

compounds

EU-OPENSCREEN complements existing resources and supports all

stages of the tool development project in an ‘open’ RI for external

researchers.

EU-OPENSCREEN – Executive Summary February 2013 Page 17

Service contract

Chemical Space

Adapted from Lipinski and Hopkins (2004) 432, 855-861

LOPAC & ACL & NIH & FMP 17/20T

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PROJECT PARTNERS

• NOR-OPENSCREEN

• Swedish Chemical Biology Consortium

• Drug Discovery and Chemical Biology

network Finnland

• Danish Chemical Biology Initiative

• Dutch Chemical Library Program

• ChemBioNet Germany

• POL-OPENSCREEN

• CZ-OPENSCREEN and Czech ChemGen

• Austrian PLACEBO

• Spanish ChemBioBank

• French Chimothèque Nationale,

PCBIS platform, FR-OPENSCREEN

• Romanian Chemical Biology Net

• Flemish Network on Chemical Biology

• Collezione Nazionale dei Composti Chimici e

Centro Screening

Current partners Associated members Coordination Centre at FMP Berlin

EU-OPENSCREEN builds on national networks in 16 European countries.

EU-OPENSCREEN – Executive Summary February 2013 Page 19 DRAFT EU-OPENSCREEN

Business Plan

Amsterdam, 20

November 2012

•Openscreen.no

• The Norwegian Chemical Biology Network, NOR-OPENSCREEN, is coordinated from UiO and includes units in Bergen, Trondheim and Tromsø: BiO/NOR-OPENSCREEN is the Norwegian partner in

University of Oslo: BiO & Chemical Synthesis and Medicinal Chemistry Group University of Bergen: Institute of Biomedicine SINTEF Materials and Chemistry Marbio/University of Tromsø: Polar Marine Bioprospecting Platform

© UiO/F. Saggio

NOR-OPENSCREEN/

ChemBioNet Norway

UiB

Marbio

SINTEF

UiO/BiO

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Chemical Biology Platform

@ Biotechnology Centre

Platform overview

The Chemical Biology High Throughput Screening platform was

established in 2008 as a service based facility

Permanent staff with continuous, solid competence in the field of chemical biology including HTS, assay development and robotics/automatisation

The facility is equipped with state-of-the-art automated screening instrumentation designed for Chemical Biology screening and to be used for both biochemical and cell based screening activities

The platform is part of the EU-OPENSCREEN, the European

Infrastructure of Open Screening Platforms for Chemical Biology

Strong Nordic collaboration

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Competence Technology Chemical Resources

In most cases user has developed an assay which has to be transferred to the platform. Examples of some assays/detection technologies AlphaScreen/AlphaLISA Enzymatic assays (absorbance/fluorescence readout) Fluoresence polarization Viability/cytotoxisity/apoptosis Calcium measurements/FLIPR Cell based assay-intracellular flow cytometry

Available a wide range of assays/ detection technologies

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Envision plate reader Filter and monochromator based detection, stacker, reagent injector for biochemical and cell based assays.

Detection platform

Molecular Devices FLIPR384 Fluorometric Imaging Plate Reader for simultaneous liquid transfer kinetic reading in 384-well plates. Cell based calcium, membrane potential and ion channel assays.

BIACORE T100 surface plasmon resonance based biosensor. Label free detection of interactions in real time.

Screening formats

96 well plate 384 well plate 1536 well plate

Reduce costs Use less reagents Faster throughput Less optimization than 1536

WORKFLOW

Detection readout

Compound administration 384 assay plates

384 compound plate

Adding assay reagents or cells to compounds

Incubation Data output

Data analysis

WORKFLOW

Detection readout

Compound administration 384 assay plates

384 compound plate

Adding assay reagents or cells to compounds

Incubation Data output

Data analysis

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Echo® Liquid Handler for Screening and OMICS -move liquid with sound

Sound waves eject precisely-sized droplets (2.5 nl) from the source liquid into a microplate.

does not use tips or nozzles—completely eliminating contact between the instrument and the liquid

Produce better-quality data, reduce costs and reagents for sample management and high-throughput plate preparation.

Enable transfer of chemical compounds, cDNA, siRNA vectors, AlphaLISA beads, proteins/Ab and crystallography reagents.

BiO chemical compound library

• ChemBioNet (17 000 compounds)

• Lopac (Library of Pharmacologically Active Compounds, 1280 compounds) )

• Prestwick (FDA approved, off-patent, 1280 compounds)

• Expansion with new compound collections (Enamine selection of 28500 compounds + Chembridge selection of 17000 compounds)

Diversity library of small moelcules collection

General purpose library for a broad range of targets

• Enriched for Potentially Bioactive Scaffolds

• Exhibit a high degree of chemical diversity

• ChemBioNet (17 000 cmpds)

• Enamine (28 500 cmpds)

• Chembridge (17 000 cmpds)

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Target and Pathway specific collections

• Bioactive compound library (1650 cmpds, Selleck Chem)

• Tocriscreen Mini, selection of biologically active compounds (1120 cmpds, Tocris)

• Target and Pathway Libraries (477 cmpds, Enzo)

Oncology collections

• Cambridge Cancer Compound Library (384 cmpds, Selleck Chem)

• Access to FIMM collection of anti-cancer drugs

• Library of 1,280 pharmacologically active compounds • Well-characterized, high-purity compounds • Represents all major target classes

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Lopac1280

The Prestwick Chemical Library®

● 1,200 small molecules, 100% marketed off-patent drugs

Greatest possible degree of “drug-likeness”

“no strings attached” compounds

● Compounds selected for:

• high chemical diversity

• high pharmacological diversity

• known bioavailability

• safety in humans

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Lopac and Prestwick: New uses for existing drugs

Used for

• Assay validation

• Hit finding

• Drug repositioning

• Drug sensitivity screen

• Start of new optimization program

New or additional value is generated from a drug by targeting diseases other than those for which it was originally intended.

Project design Project flow

HTS screen Rescreen of hit

compounds Characterisation

of hits Secondary

screen Hit-to-lead

phase Lead

optimization

User/Project owner

Assay development

User/Project owner

Chemical Biology Platform

Medicinal Chemistry

Project design Project flow

HTS screen Rescreen of hit

compounds Characterisation

of hits Secondary

screen Hit-to-lead

phase Lead

optimization

User/Project owner

Assay development

User/Project owner

Chemical Biology Platform

Medicinal Chemistry

Targeting protein protein interactions (PPI)

• Contact surfaces of PPI are mostly large and flat -contain no well defined pockets for binding of small molecules.

• Hotspots – subsets of residues that contribute most of the free energy upon binding to a protein partner or a small moelcule

• Many PPI recognise an α-helix – develop small molecules with helical mimetic scaffolds

• Generally PPI inhibitors tend to be larger in Mw and more rigid than a "typical drug"

Assay development

• Large amount of soluble recombinant protein with different tags

• Test different protein combinations, swap tags

• Obtain a robust assay with robust signal to background ratios

• Titre down bead amount and volume to reduce costs

• Transfer the bench top assay to the automated liquid handling platform

The principle of AlphaScreen

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The principle of AlphaScreen

Output

Data analysis

HTS screening (AlphaScreen) Library: ChemBioNet, 17000 compounds

Incubate

Read

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FLIPR384 Fluorometric Imaging Plate Reader

Cell-based assays of receptor/ion channel-mediated signaling processes

Measuring membrane potential changes

Evaluating changes in intracellular calcium

Real-time analysis of whole-cell signaling events

Simultaneous fluid dispensing in 384 well plate Simultaneous measurements in 384 well plate

Range = ( -3000, 20000 )

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Mini Graphs

Zebrafish: a novel way of "fishing"

• Vertebrate model with fully sequenced, annotated genome • Strong genetic, physiological and pharmacological similarity to humans • High fecundity and small size (96-well format) • Rapid development ex utero • Optical transparency (non-invasive imaging) • Only mg amounts of compounds needed; readily absorbed (skin, GI tract, gills)

Chemical Neuroscience Group @ BiO

• Develop new disease models in zebrafish for

neurological disorders (current focus, epilepsy) to create:

• genetic epilepsy models

• provide functional genomics data to systems

biology projects

• chemically-induced seizure models

• Use chemical biology in zebrafish to elucidate the

molecular mechanisms involved in epilepsy to identify:

• therapeutic targets

• therapeutic molecules

• pharmacological tools

Chemical Neuroscience Program

Compound libraries Disease Models

Toxicity Mammalian Assays

Lead Optimization & Target Identification • Chemical synthesis • Affinity Chromatography • Chemoinformatics • Pharmacology • Pharmacokinetics • Electrophysiology

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Bioassays

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