evotec DDup Special Edition

DDup SPECIAL EDITION

DRUG DISCOVERY SERVICES STATE-OF-THE-ART TECHNOLOGY PLATFORMS COMBINED WITH HIGHLY EXPERIENCED SCIENTISTS WITH EXPERTISE IN A BROAD RANGE OF THERAPEUTIC AREAS CAPABILITIES INCLUDE

New capabilities in anti-infectives Target identification & validation Screening Hit-to-lead & lead optimisation Medicinal chemistry In vivo & in vitro pharmacology Proteomics Biomarker science Compound management Integrated drug discovery

SECTIONS

1

2

3a

3b

NEW CAPABILITIES IN ANTI-INFECTIVES

TARGET IDENTIFICATION & VALIDATION

HIT IDENTIFICATION – HTS

HIT IDENTIFICATION – HCS

4

5

6

CHEMISTRY

HIGH-THROUGHPUT ADME COMPOUND PROFILING

DNA Engineering

3c

HIT IDENTIFICATION – SBDD COMPOUND MANAGEMENT

ected Cells

Transiently Transfected Cells

Non Transfected Cells

Bulk Cell Production

ant Cells

7

8

9

PROTEOMICS

CELL & PROTEIN Membrane Preparation PRODUCTION

IN VITRO PHARMACOLOGY IONProteins CHANNELS Recombinant

Cell & Reagent

10

Evotecs Screening Services

Supply

11

12

IN VIVO PHARMACOLOGY/ DMPK

EVOMAB

13

nt SERVICES INTEGRATED e m p o Devel leads

14 TRACK RECORD & CLIENTS

3

INNOVATION EFFICIENCY WITH EVOTEC A message from Evotec Chief Operating Officer, Dr Mario Polywka

Drug development is a high-reward but high-risk process, with as few as one in 10,000 new molecular entities making the successful but costly and lengthy journey from discovery to market. The pharmaceutical industry is struggling with the fall in R&D productivity and increasing demands from the regulatory authorities who demand first- and best-in-class therapies rather than me-toos.

Evotec’s drug discovery platform was established to deliver an industrialised, cutting-edge, comprehensive and unbiased infrastructure to meet the industry’s need for innovation in drug discovery. Using a strategic outsourcing partner like Evotec to drive Innovation Efficiency, reduces the risk and increases the chance of success in drug discovery without the need for the industry to invest in fixed cost structures.

Evotec’s portfolio of capabilities includes target identification and validation, compound management, screening, hit-to-lead and lead optimisation medicinal chemistry, in vivo and in vitro pharmacology, prote­ omics and biomarker science. Work-

ing with Evotec, our partners gain access to an industry-leading team of scientists with years of experience in the Pharma and biotech industry.

This experience effectively marries target and disease biology expertise with a world-class technology infrastructure. These scientists have successfully delivered clinical candidates in their past careers but more importantly they continue to do so at Evotec for our clients. The intellectual input of Evotec’s scientists is one of the pillar’s supporting Evotec’s business model. The most visible sign of our scientist’s ability to deliver innovative solutions is the number of customer patents on which Evotec scientists are named inventors. This number grows yearly and currently exceeds 200 and covers many therapeutic areas and target classes.

Evotec is the partner of choice for stand-alone services which are charged on a purely fee-for-service basis. Alternatively, we can offer holistic, fully integrated drug discovery solutions through a va­riety of commercial structures. Our aim is to meet your requirements for the drug discovery solution you are seeking both scientifically and commercially. Our track record is second to none as testified by the number of projects that have moved through to the clinic and the strong portfolio of satisfied partners we have worked with through the years.

Evotec has been involved in more than 250 partnerships since its inception in 1993 and has delivered more than 30 pre-clinical candidates and 20 clinical candidates both in partner­ ships and within its own proprietary drug discovery efforts.

This document showcases the stateof-the-art offerings of Evotec to address the challenges of Innovation Efficiency within our industry. In particular with this Drug Discovery Services update we introduce you to our new capabilities in anti-infective research. Earlier this year we acquired Euprotec, a UK based company with one of the world leading platforms for infectious disease research. This acquisition gives us access to StrainBank, a unique collection of clinical isolates useful in establishing the activity profiles of compounds against bacterial and fungal pathogens as well as key capabilities in microbiology, pharmacology and DMPK. Of critical importance is the key team of scientists from Euprotec that have now become a part of the Evotec family.

Please use this document to define how we can help you and contact us for the most innovation- and cost-efficient solution to your drug discovery requirements.

Yours sincerely Mario Polywka

SECTION 1

NEW CAPABILITIES

IN ANTI-INFECTIVE RESEARCH

Lloyd Payne Senior Vice President Anti-infectives Operations

Evotec’s specialist group in the infectious disease therapeutic area boasts state-of-the-art microbiology facilities including a unique and highly characterised strain bank. The group was established in 2008 as Euprotec and was acquired by Evotec in May 2014. The group provides bespoke anti-infective drug discovery and development services to a growing number of global clients and brings to Evotec an established track record in collaborating to discover and develop therapies and vaccines to treat and prevent serious and lifethreatening infections resulting from multi-drug resistant bacteria and fungi including MRSA, Pseudomonas, and Clostridium difficile.

In 2012, Euprotec was named the Bionow “Biomedical Service Provider“ of the Year in recognition of delivering scientific excellence and the very best in customer service to all of its clients. The group’s offering is fully integrated with the wider discovery platform at Evotec to enable us to offer either standalone microbiology services or conduct fully integrated anti-infective drug discovery projects. Key capabilities of Evotec’s microbiology group fall into the four areas of EvostrAInTM, Microbiology, Pharmacology and ADME/PK/PD as follows:

extensive collection, EvostrAInTM, or strains provided by our collaborators. This includes the ability to conduct whole-cell screening for antimicrobial activity for hit identification, MIC, MBC/MFC, timekill and PAE studies using single or combination agents, hollow fibre PK/PD or bioreactor human cell systems for detailed profiling for characterisation of novel antiinfective agents and compound/ drug combination studies for assessment of synergistic, antagonistic and additive effects. Bespoke methods for susceptibility profiling can be developed for testing novel EvostrAInTM agents where standardized methods TM EvostrAIn is a unique collection may not be appropriate. In parallel of clinical isolates that can be used to mechanism of action and resistance establish the activity profile of lead frequency assays can be performed. compounds and candidates. A key PHARMACOLOGY feature is that the isolates are highly characterised and, in many cases, We specialise in assessing the mechanisms of resistance defined. efficacy of lead and candidate Our strain collection contains an compounds in highly relevant extensive range of geographically and validated disease models. Our diverse human bacterial and fungal extensive range of established pathogens that cover isolates models are well-suited to the susceptible and resistant to current develop­ment of multiple classes of agent including small molecules, antimicrobial drugs. natural products, peptides, MICROBIOLOGY antibodies, other biologics and We employ industry-standard vaccines. We provide a highly methods including CLSI, bespoke service, customising studEUCAST and BSAC to test ies to meet the exact requirements compounds for antimicrobial activ- based on programme needs and ity against organisms from our parameters/endpoints of interest.

5

EvostrAInTM BACTERIA: Gram positive pathogens

BACTERIA: Gram negative pathogens

FUNGI

Staphylococcus aureus including MRSA, VISA and VRSA strains

E. coli including Extended Beta lactamase producing strains

Aspergillus spp. (including strains resistant to azoles, polyenes and echinocandins with known mechanisms of resistance)

硫-Haemolytic streptococci groups A, B, C and G

Klebsiella pneumoniae Carba足penemase producing strains (KPCs & MDR)

Candida spp. (including strains resistant to azoles, polyenes and echinocandins with known mechanisms of resistance)

Streptococcus pneumoniae (in足cluding penicillin, macrolide, fluoroquinolone, cephalosporin and MDRSP resistant strains)

Acinetobacter baumannii including MDRAB

Mucorales

Vancomycin Resistant Enterococci (VRE)

Pseudomonas spp. including multi-resistant strains

Cryptococcus

Bacillus species

Haemophilus influenzae

Dermatophytes including Malassezia spp and Trichophyton spp

Listeria species

Bacteroides spp.

Fusarium

Corynebacterium and Propioni足bacterium species

Neisseria gonorrhoeae and N. meningitidis

Protozoa Acanthamoeba spp

Clostridium difficile (multiple ribotypes including 012, 027 and 078)

Intestinal pathogens: Vibrio spp, Campylobacter spp, Salmonella spp, Shigella spp, Yersinia spp.

Other Clostridia (including C. perfringens)

Legionella spp.

Mycobacterium

For agent efficacy assessment against bacterial and fungal infection our models can address different sites of infection (localised and systemic) by Gram-positive (including Staphylococcus aureus, Streptococcus pneumoniae, and others), Gram-negative (including Pseudomonas aeruginosa, Escherichia coli, Acinetobacter baumanii and others), anaerobes (including Clostridium difficile) or fungal species (including Candida sp., Aspergillus sp., Mucorales, Malassezia sp.).

ADME/PK Our existing extensive ADME/PK offering is now enhanced with the ability to address anti-infective specific capabilities. In addition to high quality pharmacokinetic profiling and bioanalysis these include: a) Bioassay/bioactivity drug quantification b) Development of bespoke PK models and assays with drug quantification at multiple sites c) Translation of PK data into

efficacy models to optimise outcomes d) Tolerability assessment utilising multiple endpoints e) Early assessment of cytotoxicity potential against multiple mammalian cell lines.

In particular, Euprotec specialises in PK/PD profiling and modelling in multiple disease models.

SECTION 2

TARGET IDENTIFICATION AND VALIDATION World-class ex vivo imaging technology platform using tissue sections to study cellular and molecular events Phenotypic screening of complex cellular systems for hit identifi足cation Target deconvolution using proteomics GENE EXPRESSION PROFILING

Andreas Scheel Executive Vice President In vitro Biology In vitro Pharmacology

Changes in gene expression, as result of a disease state or as a consequence of drug action, give invaluable insights into biological systems. From early target iden足 tification to mode of action studies and biomarker identification the analysis of gene expression data supports the drug discovery processes. Our capabilities include: Design and execution of transcriptomics studies in vitro or in vivo including rigorous quality control Bioinformatics analysis to determine differential expression and generate annotated gene lists Cluster analysis to identify groups of regulated genes Data interpretation turning raw data into meaningful biological and pharmacological information

OFFERING Evotec has established a worldleading platform to support the discovery and validation of innovative disease-modifying targets across different therapeutic areas.

Our offering comprises: Differential expression studies followed by bioinformatics-driven data mining and hypothesis building Knock-down and over-expression IN VITRO TARGET studies both in vitro and in vivo MODULATION CAPABILITIES as well as access to relevant in The powerful combination of vivo disease models siRNA and disease-related cellular

models coupled with a variety of read-outs including high-content screening are designed to provide an accelerated solution to target identification and validation. Our offering includes: Screening of siRNA libraries for target identification Validation of specific targets in various cellular models using disease-relevant read-outs Evotec has built substantial experience in this area and routinely achieves knock-down effi足 ciencies of at least 80%. Additionally, expertise in working with nontraditional cellular models such as co-cultures and cellular aggregates is available.

IN VIVO TARGET VALIDATION Evotec has disease models in a number of areas such as dia足betes, diabetic complications, kidney diseases, neurodegeneration, oncology, inflammation and pain that can be used for target validation. Approaches utilised include: Application of target-selective small molecules or biologics in acute or sub-chronic in vivo studies in rodents Virus-based target in vivo knockdown and over-expression techniques

7

A variety of read-outs have been established that allow the investigation of target modulation. Such studies are run on a routine basis and additional read-outs are developed on a continuous basis.

Classical pharmacodynamic and efficacy read-outs such as blood glucose and HbA1c for diabetes, behavioural read-outs for pain and neurodegeneration A world-class ex vivo histology and immunohistochemistry platform investigating cellular and molecular markers in tissue sections prepared from treated animals

Identification of new mechanisms to treat kidney disease: Phenotypic screening of conditionally immortalised human podocytes

USE OF PHENOTYPIC ASSAYS TO IDENTIFY NEW DISEASE RELEVANT TARGETS AND PATHWAYS The ability to study compound effects in a disease-relevant cell type in a target-unbiased way offers enormous potential and has thus become increasingly popular in recent years, with more complex and disease-relevant cellular systems becoming available for higher throughput assays.

High-Content Screening Evotec has a world-class highcontent screening platform that is increasingly used for hit identification through phenotypic screening >15 years know-how in the design and implementation of phenotypic assays using cell lines but also including complex cellular systems utilising primary cells and stem cells in the area of neurodegeneration, inflammation, metaEvotec has in-depth expertise in bolic disease and oncology A small molecule compound the area of phenotypic screening library of 400,000 compounds and target deconvolution.

Proteomics-based target deconvolution Evotec has established a mass spectrometry-based approach to ex­perimentally determine the binding partner(s) of a lead compound in a relevant cellular context: Metabolic or chemical labelling Optimised linker chemistry capabilities Determination of Kd values for each putative binding partner Combine with post-translational events to understand global cellular effects

Overview of Evotec capabilities and expertise in target identification and validation across different disease areas

METABOLIC DISEASE

ONCOLOGY

CNS, NEURODEGENERATION, PAIN

+++

n.a.

+++

++

++

++

Cellular / ex vivo tissue imaging

+++

++

+++

In vivo efficacy studies

+++

+

+++

Differential gene expression siRNA / RNAi / overexpression

SECTION 3A

HIT IDENTIFICATION

HIGH-THROUGHPUT SCREENING OFFERING Evotec has a long history in highthroughput screening (“HTS”) utilising large client compound collections and also our own 400,000 compound collection. Evotec’s screening platform is differentiated through integrating a variety of detection technologies with high-throughput capabilities. This technology platform allows for assay miniaturisation and use of multiple read-out parameters to enable the reduction of false positives and negatives. Evotec’s technology platform

Dirk Ullmann Executive Vice President Lead Discovery

includes nuclear magnetic resonance spectrometry (“NMR”), surface plasmon resonance spectrometry (“SPR”), high-content screening (“HCS”) and high-throughput mass spectrometry based screening (“HTMS”).

OVERVIEW SCREENING CAPABILITIES AT EVOTEC Evotec selects the most suitable screening strategy for your target – our approach is to let the target dictate the screening strategy and not the other way around.

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Screening platforms at Evotec

*Available RapidFire MS capacity allows for up to 10,000 data points per day

EVOSCREEN MARK II

EVOSCREEN MARK III

384-HTS SYSTEM

RADIOMETRIC PLATFORM

No. of Systems

1

1

3

semi-automated benchtop devices

Assay Volume

1µl/well

5-10µl/well

20-50µl/well

20-50µl/well

2080-well

1536-well

384-well

96 and 384-well

Formats

Biochemical

Cellular + Biochemical

Cellular + Biochemical

Cellular + Biochemical

Readers

Insight™ FCS+plus reader

PE EnVision

FLIPRTetra, PE EnVision, Tecan Safire, RapidFire MS

PE TopCount (2) PE MicroBeta2 (3)

Assay principles

Binding Enzyme activity

Binding Enzyme activity Reporter gene

Binding Enzyme activity Ca2+ flux Second messenger Membrane potential

Radioligand binding FlashPlate SPA Filtration

FCS+plus

Fluorescence, Luminescence, Absorption, Alpha screen

Fluorescence, Luminescence, Absorption, Alpha screen, Mass spectrometry*

up to 100,000 / day

up to 40,000 / day

Plates

Detection

Throughput

up to 100,000 / day

Isotopes: H, 35S, 14C, 32P, 33P, 59 Fe, 125I

3

up to 25,000 / day

Assay development and HTS track record ASSAY DEVELOPMENT Others 109

SCREENING Kinase 74 Phosphatase 9

Others 20

Ion Channel 19 Protease 16

Protein Protein 23 ADMET 5

Kinase 19

NHR 9 Protease 67 ADMET 25 Ion Channel 54 >400 assays developed Success rate >95% High percentage of cellular assays

Evotec has twenty years of experience in assay development; in particular in HTS, but also to support hit-to-lead & lead optimisation (H2L/LO) programmes.

Our experience includes: A broad portfolio of assays

Enzyme other 79 GPCR 59

GPCR 43 >220 uHTS screens Specific hits with EC50<25μM identified in most screens

covering the major target classes (see chart) New target classes in the field of epigenetics and protein-protein interaction

High numbers of unique assays developed as no literature

precedence was available Assay read-outs covering a wide range of technologies including both biochemical and cellular assays

Assays using label-free and biophysical methods

SECTION 3A

For structure-based approaches to drug discovery, Evotec has a 20,000 fragment set available for biochemical screening and a 3,000 fragment set specifically selected for NMR and SPR screening. Evotec screens customer libraries of any size, virtual screening derived hits and also smaller, more target-directed libraries.

THE OPTIMUM SCREENING COLLECTION Our library contains 400,000 compounds consisting of 70,000 proprietary Evotec compounds and a set of 330,000 maximally diverse “islands”.

Screening process

Diverse islands 330,000

SCREENING PROCESS The integration and connection of the different stages of the screening process from assay development to automation to compound management/tracking and finally data processing are essential. The Evotec screening team manages this complexity on a daily basis to Evotec’s screening library is differ- deliver high-quality data on time to entiated through its quality, diver- our clients. sity and novelty and has a successful track record of delivering potent In Evotec’s view the success of a and attractive compounds as star­ screening project is determined by: Target selection ting points for medicinal chemistry Right strategy for target expresoptimisation. sion to address specific sites of interest Robust and sensitive assay system Well-selected compound collection for screening Evotec fragment 20,000 Reliable screening process Careful characterisation screening Evotec proprietary 70,000 hits in orthogonal test systems

11

Distribution of calculated properties in Evotec’s discovery screening library Molecular weight distribution

0

CLogP distribution

100 200 300 400 500 600 700 800 900 Molecular weight

-5.0

Rotatable bond count

2

4

6

0

2.5 CLogP

5.0

7.5

10

0

H-Bond acceptor distribution

8 10 12 14 16 18 20 Rotable bonds

1

DATA HANDLING Evotec’s data analysis platform, Aplus, is suited to support all screening processes from HTS including multivariate analysis

Reagent production and assay development

-2.5

TPSA (Topological polar surface area)

MINIATURISATION

2

3

4

H-Bond donor distribution

5 6 7 8 9 10 11 12 H-bond acceptors

for HCS to SPR and also HTMS. This software facilitates the integration of all techniques into one analysis platform and streamlines the process flows. For customer

Adaptation on screening platform

MARKER LIBRARY SCREEN

20 40 60 80 100 120 140 160 180 TPSA

1

2

3 4 5 H-bond donors

6

7

reporting, Evotec is flexible to meet customer needs in data formats for upload in different environments.

Primary HTS screen n=1

DATA STATISTICS A+

Primary hits (3sigma threshold)

HIT CONFIRMATION SCREEN N=3

Identification of clusters & singletons

PRELIMINARY SAR

Specific hits with confirmed conc. dependent activity

TEST IN ORTHOGONAL ASSAY

Hits with conc. dependent activity

11PT DRCS N=2 + LC/MS CHECK

Confirmed hits

SECTION 3B

HIT IDENTIFICATION HIGH-CONTENT SCREENING cell derived or primary cells from appropriate hosts. This approach is a core part of Evotec’s efforts to identify novel targets and drugs with disease-modifying potential.

Ina Sternberger Group Leader In vitro Pharmacology

OFFERING Evotec specialises in the development and implementation of high-content based assays for high-throughput screening and compound validation to probe the pathophysiological environment. High-content-screening (“HCS”) is routinely used throughout the hit identification cascade from target validation to mechanistic studies. The assays developed at Evotec encompass both a target centric as well as phenotypic approach with emphasis on tissue samples, stem

Evotec’s HCS expertise is built around the OPERA® platform where an essential understanding of the mechanics and image analysis tools, gathered over a period of more than 15 years in the development of the platform, flows into the hit identification process. The platform is completed with a powerful data management system and powerful data analysis tools including multifactorial analysis. Based on this expertise, Evotec has performed many high throughput screens of up to 350,000 compounds and developed over 30 assays to study mode of action of compounds and targets in a multitude of cellular backgrounds.

Accessing Evotec’s HCS platform offers a road to novel target space achieved via an array of diseaserelated secondary models post-HTS coupled to target deconvolution via the application of bioinformatics, siRNA, gene expression and finally mass spectrometry based proteomic approaches.

TARGET/ASSAY EXPERTISE The nature of HCS describes primarily a target agnostic approach

PHENOTYPES Proliferation Cell cycle analysis Cell death/necrosis/apoptosis Differentiation markers DNA damage Cytoskeleton rearrangements Neurite outgrowth Mitochondrial mass Endocytosis Synapse density Neurodegeneration/-protection Intracellular aggregation Post-translation modification

measuring phenotypic/morpho­ logical changes associated with target engagement.

The use of phenotypic screening provides Evotec’s clients with a whole new realm of drug discovery opportunities that otherwise could not be addressed. This includes experienced multi-disciplinary teams that build strategies for hit follow up towards identifying the molecular mechanism of action of a potential drug. Other assay formats that are routinely developed on Evotec’s HCS platform are designed to investigate subtle or rare subcellular events. The table shows examples of cellular events that Evotec has experience with on the OPERA® platform.

DISEASE AREA EXPERTISE HCS at Evotec is now a common element in many drug disco­ very programmes across a variety of therapeutic areas. At present, the main focus of HCS is in the area of oncology, kidney disease, diabetes, pain,

inflammation and neurodegeneration. Here the strengths of HCS are combined with regenerative medicine concepts to probe and discover new treatment paradigms such as beta cell regeneration for diabetes and podocyte protection for diabetic

neuropathy. Applying stem cell technology in phenotypic and highcontent screening is also an area of active research at Evotec where we are committed to delivering new treatment paradigms for ALS and other degenerative diseases.

Examples of track record

TARGET/PHENOTYPE

READOUT

DRUG ID PHASE

Transporter

Internalisation

HTS 250K

Fibrosis

αSma expression/actin alignment

HTS 50K

Beta cell proliferation

Insulin expression coupled to BrdU stain

HTS 10K Biologics

Anchorage independent growth

Target validation

siRNA screen

Diabetic Neuropathy

Actin rearrangement

Known drug collection

Striatal neuron development

DARP32 positive cells

Protocol development

Neurodegeneration/ALS

Neurite outgrowth in co-culture system

10K HTS + known drugs

Transporter

Binding of fluorescent ligand

HTS of 100K

Huntington’s protein aggregation

Number of aggregates

Known drug collection

NFκB translocation

Nuclear accumulation in HUVECs

Lead optimisation

PTM enzyme in AID

NET formation in primary human neutrophils

Compound validation

DNA repair/oncology

DNA damage response in tumor cell line

HTS 50K

Autophagy

LC3 accumulation in autophagosomes

Pilot screen

Epigenetics

Protein methylation

Hit follow-up

SECTION 3C

HIT IDENTIFICATION

STRUCTURE-BASED DRUG DESIGN (SBDD) Guided by high-quality data, SBDD at Evotec distils information into hypothesis-driven medicinal chemistry. By minimising unnecessary and wasteful compound synthesis, productivity is increased and chemistry teams are able to focus on an efficient path to the final drug.

tification of new starting points for SBDD programmes. Evotec offers both protein-observed NMR assays monitoring chemical shift perturbations (SAR-byNMR) as well as ligand-detected NMR assays (Saturation Transfer Difference (“STD”) NMR and Water LOGSY) for hit identiEvotec has a comprehensive SBDD fication. Hence, a broad range platform comprising state-of-the- of target proteins amenable to art capabilities in: NMR screening technique can be Computational chemistry covered.

Protein engineering X-ray crystallography Evotec has employed its NMR Biophysical screening tools such screening platform on more than as NMR, SPR, MS and ITC 30 protein targets covering a whole

John Barker Vice President Structural Biology

FRAGMENT COLLECTIONS We employ our collection of 20,000 fragments for biochemical, high concentration and SPR screening and a 3,000 fragment set for NMR screening all of which are optimally designed for quality, diversity and solubility at high concentrations to provide tractable starting points for optimisation within SBDD projects.

The collection integrity is maintained following validated processes used in our HTS collection to maintain quality, diversity and novelty.

OFFERING The SBDD platform at Evotec includes a comprehensive fragmentbased drug discovery (“FBDD”) platform. Sensitive screening techniques, such as the FCS+plus (proprietary to Evotec) or NMR, SPR or mass spectrometry (MS), combined with specifically selected fragment compound collections, identify diverse, active NMR SCREENING AND low molecular weight fragments HIT QUALIFICATION providing multiple highly efficient NMR screening as part of the starting points for chemical optimi- Evotec FBDD platform is an sation. alternative option for the iden-

H, 15N-Trosy Spectra (37kDa kinase, H, 15N-labelled)

1 2

apo-protein Protein + fragment

range of target classes from frequently studied enzymes (like kinases or proteases) over nucleotide binding proteins or epi­ genetic targets, to proteinprotein inter­ action targets, even receptor domains. NMR also adds con­ siderable value to biochemical screening campaigns by validating hit compounds for direct binding to the target protein and ultimately correlating this information to structure data, thereby enabling or facilitating SBDD programmes.

Aurora-B kinase inhibitor was developed alongside Roche with extensive structure-based design. Evotec employs multiple structure-based analytical tools to help prioritise compound ideas, alongside collaborative tools for communication with medicinal chemists

bilised target proteins. As the throughput is sufficiently high, we are able to screen both fragment sets thus increasing the chance to find bona fide binders of the target.

Independent of the primary screening strategy applied the assay techniques available at Evotec allow qualification of hits using alternative read-outs. Here we typically combine the above mentioned SURFACE PLASMON approaches with orthogonal assays RESONANCE (“SPR”) SCREENand technologies like Isothermal ING AND HIT QUALIFICATION Titration Calorimetry (“ITC”), Another powerful tool for study- Differential Scanning Fluorimetry ing biomolecular interactions in (“DSF”) and others. a sensitive and label-free manner STRUCTURAL BIOLOGY in real-time is SPR. While commonly used at later stages of Evotec is located within a few drug discovery projects to qualify miles of the UK’s third generation hit compounds with regards to synchrotron and coupled with our binding kinetics, the instrumenta- own in-house X-ray source offers the tion available at Evotec also allows ideal infrastructure for rapid cycle for primary screening of frag- times in X-ray crystallography. The ments. Direct binding assays run Structural biology team provides on the Biacore™ 4000 screening solutions to novel structure elucidevice enable the identification of dation of drug targets and optimifragments interacting with immo- sation of crystal systems to support

industrialization of protein:ligand complex crystallography. Delivery of protein complexes is fine tuned across a project’s lifetime to maintain the highest possible resolution while meeting the demanding cycle times of modern dis­covery. Experience covers the major target families with recent focus on crystallo­ graphy of membrane proteins.

The team is co-located with the medicinal chemistry and computational chemistry teams to maxi­ mise interaction across integrated discovery programmes. The use of 3D virtual reality has provided the chemistry team with the ideal environment to understand and effectively mine structural information to focus synthetic strategies and rapidly progress optimisation.

Working with innovative computational scoring techniques, Evotec is experienced in maximising the value of structural information, providing foundation for rational design.

SECTION 4

COMPOUND MANAGEMENT

Scott Snyder Executive Vice President Compound Management

OFFERING Evotec is uniquely qualified in delivering its customers Compound and library management needs through over a decade of being the world’s leading compound management operation. Evotec expanded its operational presence in 2013 by adding a second location in Branford CT which is strategically positioned to support the strong presence of Evotec’s drug discovery collaborators along the East Coast of the United States. Our service offering includes: Cost effective compound identification, selection, and procurement High-throughput compound analysis Multi-format plating and reformatting Inert atmosphere and low temperature storage and processing Fast order fulfilment and worldwide delivery Multi-site disaster recovery and business continuity “In-sourcing” of Evotec compound management staff to client sites Our compound management experience, primarily from the large Pharma and government sector means that Evotec understands its customers needs and deadlines. Each client has a dedicated Project Manager who provides a single point of contact to insure an open line of communication is always maintained.

COMPOUND IDENTIFICATION, SELECTION, AND PROCUREMENT Evotec has extensive experience in the identification, selection and procurement of sub-collections of compounds possessing very specific (including commercial, physicochemical or bioactivity-related) properties. Evotec has the capa­ bility to apply compound identification tools as required by the client.

Once compounds are identified for purchase, our Project Management group consults with selected suppliers to negotiate terms on behalf of our client requirements, e.g. price, purity and format.

Evotec purchases compounds on a regular basis for a number of clients; therefore, the client can benefit from our vendor relations and volume discounts.

COMPOUND QUALITY CONTROL Evotec typically uses a three-part incoming Quality Control (“QC”) process comprised of weight, solubility and LC-MS purity and identity to ensure that all collections start with high-quality compounds.

There are two Waters/Micromass 8-channel LC-MS systems, each with digital UV, evaporative light

17

scattering (ELSD) and an LCT TOF mass spectrometer. These are high-throughput instruments with a capacity of 500,000 injections per year. Evotec also offers ultra-high performance liquid chromatography with ELSD and electrospray mass spectrometry (UPLC-MS) method for quantitative determination of compounds in solution in DMSO. COMPOUND STORAGE Evotec has invested in industryleading automated storage systems and supporting informatics. Storage capabilities support vials, tubes, plates and bulk materials at controlled temperatures of -80°C, -20°C (inert atmosphere) and ambient. Actual storage conditions are dictated by sample type, containers and length of storage.

Automated tube storage systems are equipped with high-speed pickers for fast arraying of tubes to support large plating campaigns and rapid follow-up. Evotec can also prepare a disaster recovery set of the client’s collection for storage at one of our other compound management facilities. This set can be readily accessed and processed for distribution at the client’s request.

COMPOUND DISPENSATION AND REFORMATTING DMSO solutions Evotec has the capability for automated arraying of large numbers of compounds in DMSO between a large variety of containers suitable for distribution or storage.

Our extensive liquid handling resources cover the widest range of possible volumes.

High-volume end of the range are the eight channel pipettors, Tecan and Sias systems that can dispense and transfer 5 uL to multi-milliliters in vials and plates.

Middle of the range are the Agilent plate replicators (formerly VPrep and Bravo) that can dispense from 1 uL to 2 mL to and from 96, 384 and 1536 SBS format arrays Low end of the volume range is the LabCyte Echo 550 that acoustically dispenses volumes below 1 uL using multiples of 2.5 nL drops of solution Neat compound dispensation Evotec has the capability to transfer neat samples via manual weighing, Volatile Solvent Transfer (“VST”) or Solvent Transfer methods.

SECTION 4

COMPOUND SHIPMENT Our team has extensive experience in shipping chemicals, research compounds, controlled substances and reagents. Evotec delivers and receives compounds in all formats, both domestically and overseas (including, but not limited to, North America, Europe and

Asia). We are well-versed in documentation requirements, tariff classification and import/export customs requirements. We have the ability to facilitate the import/ export processes through the use of brokers as required by the Client.

Experts in Sample Logistics

> 44 million samples delivered to 377 recipients in 18 countries

DATA MANAGEMENT Evotec currently uses both a proprietary Compound Inventory and Tracking (“ComIT”) software system and the industry leading commercial application (Titian Mosaic) to manage, record and report all manipulations of client samples. These systems oper-

19

ate independent of each other at different sites but both deliver end-to-end functionality and traceability.

In support of our disaster reco­ very and business continuity plans Evotec employs i365 Data Backup and Recovery Solution to store incremental backups on a nightly basis and full server backups on a weekly basis. Data are transferred to two offsite locations automatically.

All data transmissions are encrypted and password protected at all times for security and confidentiality.

our output. Our process enables us to find defects early in the process and our procedures ensure corrective action is taken whenever a defect occurs, with the goal being to QUALITY MANAGEMENT implement corrections with miniEvotec operates under the mal impact to our final deliverables.

ISO9001:2008 Quality ManageIN-SOURCING AND ment and internal auditing princiCONSULTING SERVICES ples but is not formally ISO9000 certified. We have well-defined and If a prospective client decides to documented procedures to moni- retain compound management tor and ensure the consistency of services within their operation then

EVOTEC TRACK RECORD Build and operate both private and public screening libraries since 2004 Largest individual library consists of ~490,000 compounds Over 25 client collaborations working to individual specifications > 44 million samples shipped to 377 recipients in 18 countries Over 4 million samples acquired and processed for clients Experience with 223 compound global suppliers

Evotec also offers both in-sourcing and/or consulting services. In these instances, Evotec reviews requirements with our clients and design solutions including need identification, process design, facility layout, process workflow, vendor qualification and selection, and even complete staffing and management of onsite services.

Our in-sourcing program provides scientific staffing specifically designed to give clients the “nonpermanent” workforce needed for anywhere from one year to multiyear partnerships. We hire, train and manage our employees to perform analytical scientific programs at client sites, using their quality systems. This advanced staffing model provides a nonpermanent, long-term and costeffective way to meet compound management staffing needs.

SECTION 5

CHEMISTRY

Dave Hallett Executive Vice President Chemistry Discovery Chemistry

OFFERING Small molecule drug discovery requires a deep understanding of biological context coupled to know­ ledge and experience of the property space required to produce safe, efficacious drugs. Over the past 20 years, Evotec has developed expertise in numerous therapeutic areas including diseases of the central nervous system (“CNS”), oncology, diabetes and metabolic diseases, pain, inflammation and anti-infectives.

We understand that each partner has different needs, internal capabilities and capacities. Evotec prides itself in being able to provide flexible, innovative and efficient solutions. Evotec’s experienced scientists can support your aspirations in a variety of ways.

The capabilities listed below can be accessed individually where required or brought together to form complete project solutions: Cutting-edge molecular design Outstanding molecular modelling and computational chemistry tools Rapid synthetic execution In vitro ADMET, rodent PK and bioanalysis Structural biology Expert advice on overall project strategy Developing strategy for and securing IP protection Project management

sis is concerned, speed is paramount. The faster our teams can deliver molecules into bioassays, the faster the project teams can obtain new knowledge.

Our approach is to employ, educate and retain talented chemists and to enable them with the latest technology and equipment.

More than 60% of our chemists are educated to PhD level >40% of our chemists have >8 years experience at major pharmaceutical and biotech companies prior to joining Evotec These facets enable Evotec to provide superior synthetic chemistry and problem-solving capabilities to our partners and to deliver compounds in a shorter timeframe than our competitors.

Good molecular design often leads to complex syntheses (e.g. densely functionalised heterocycles, multiple chiral centres). Projects are successfully executed at Evotec that involve highly challenging syntheses and isolation procedures, for example: Natural products Steroids Macrocycles Nucleoside synthesis Synthetic execution is one of the Sensitive pro-drugs most cost- and time-consuming parts of the early discovery process.

We understand that where syntheSYNTHESIS At the foundation of Evotec’s medicinal chemistry group is a talented and industry-experienced team of >150 synthetic organic chemists. Their skills are utilised during medicinal chemistrydriven projects, focused-library preparation and when preparing chemical building blocks or literature compounds.

21

The origin of the chemistry department was in asymmetric synthesis and Evotec routinely develops enantioselective synthetic methods and has a full range of chiral separation technologies including SFC to deliver single isomers.

MEDICINAL CHEMISTRY

scientists has made significant contributions to discovery projects throughout their careers: >90 development candidates Named inventors and authors on >750 patents and publications Our scientists are drawn from a variety of backgrounds and have been successful in all major therapeutic areas and target classes.

Our teams are fluent and expert in both protein structure-guided and ligand-based design.

Evotec has a powerful core of experienced, industry-seasoned drug hunters who drive the molecular design process. Collectively, this co-located group of medicinal chemists, computational chemists, The focus of our medicinal structural biologists and DMPK chemistry teams is on high-quality

plC50

Series A plc50 5.45 LLE 3.16 clogD 2.3

4

5

Series B plc50 5.58 LLE 2.9 clogD 2.7

Series A analogue plc50 6.8 LLE 5.3 clogD 1.5

6

Series C plc50 7.19 LLE 3.78 clogD 3.4

7 Series B analogue plc50 6.62 LLE 4.1 clogD 2.5

8

Series C analogue plc50 6.97 LLE 4.65 clogD 2.3

9

Series C plc50 7.29 LLE 4.02 clogD 3.3

LiPE6

0

1

2

3

An example of multiparameter optimisation.

Evotec’s design teams simultaneously improve physicochemical properties as well as biological activity. The plot above (-log10IC50 versus calculated LogD) highlights three different chemotypes from the same project. Over 2 iterations, the arrows indicate the improvements made to each series as seen from the improved Lipophilic Ligand Efficiency (“LLE”) scores

LiPE3

LiPE5

4

LiPE2

design and crisp decision making.

The process at Evotec is enabled by widespread availability of visualisation, analysis and design software tools. Our medicinal chemists target analogues within defined property space to answer specific structure-property and structure-activity questions – the emphasis is always on making the right compounds with optimisation of biological activity and a properties in parallel. We firmly believe that the co-location and interaction of synthetic and medicinal chemistry, computational science, structural biology and DMPK enhances the overall design, analysis and communication process: More learning cycles per unit time = more knowledge and experience = better quality output More informed decisions = less waste

LiPE4

5

cLogD

SECTION 5

COMPUTATIONAL CHEMISTRY Evotec’s Computational Chemistry group comprises 10 PhD molecular modelers working with both commercial and in-house developed software tools.

The members of this group come from diverse backgrounds and through close interactions within project teams they provide unique insights and input across the whole discovery process: Virtual screening (X-ray structure, homology and ligand-based) Multi-parameter (“QED”) analysis of private, commercial and virtual compound sets Construction and refinement of structure-activity and structureproperty relationships Proprietary suite of GPCR modelling tools Execution and graphical visualisation of complex quantummechanical calculations which highlight to Evotec’s medicinal

chemists the components that contribute to binding in proteinligand complexes (Fragment molecular orbital analysis) Creation of electronic workflows for our teams (e.g. Knime) to provide our chemists with desktop access to complex computational methods

Routine testing (e.g. weekly) and expert interpretation of data within integrated medicinal chemistry teams

Evotec’s combination of routine in vitro assays and our flexibility to address your individual requirements enables us to offer a truly bespoke service – experienced DRUG METABOLISM AND DMPK scientists delivering data of PHARMACOKINETICS (“PK”) the highest quality coupled with a Evotec offers a comprehensive and cost effectiveness afforded by highly flexible range of in vitro ADME automated procedures. and PK services designed to cover STRUCTURAL BIOLOGY the majority of activities in the screen-to-candidate phase. The Evotec’s crystallography group scientific expertise and assays are comprises a team of 8 skilled accessed in a variety of ways by our crystallographers together with state-of-the-art-infrastructure to clients and project teams: Stand-alone profiling support protein production and — Automated, medium through- purification to the high standard put assays (100s of compounds required for this biophysical techper week) nique. The groups’ collective experi— Tailored packages designed to ence covers the major target families address a specific compound or with a recent focus on crystallogproblem raphy of membrane proteins. The

23

A summary of our current capabilities

N TIO

RI N

AB

TIO

SO

BU

RP

ST N ET

IO

IS

CR

OL M

EX

B TA

Evotec’s expertise in medicinal chemistry has enabled it to successfully execute collaborations with a broad range of partners from academic institutions through to large pharmaceutical companies.

ME

group benefits from a unique geography; co-located with the medicinal chemistry and computational chemistry groups at Evotec as well as being <5 miles from the UK’s national synchrotron (Diamond).

When discovery projects are fortunate to be structurally-enabled, Evotec’s crystallographers form an integral part of the molecular design team. As a result of this, they are acutely aware of the rapid cycle times required if novel protein-ligand complexes are to make a meaningful contribution to the design process. Delivery of protein complexes is flexibly allocated across a project’s lifetime to maintain the highest possible resolution while meeting these demands.

METABOLISM Microsomal stability Hepatocyte stability Plasma stability Metabolite identification CYP450 inhibition CYP450 induction

DISTRIBUTION Plasma protein binding Serum binding Brain tissue binding Blood:Plasma partitioning DI

ABSORPTION Aqueous solubility PAMPA Caco-2 MDCK-mdr1 SIF stability LogD (Shake Flask) Chromatographic LogP

EXCRETION Rodent PK IV, PO, IP, SC, IM Metabowl excretion Bile duct cannulation

EVOTEC TRACK RECORD Over the last 15 years, Evotec chemists have supported >125 hit-to-lead and/or lead optimisation projects Evotec scientists are named inventors on >200 client patents and have helped identify >30 pre-clinical candidates Our project teams have made major contributions to the identification of >20 compounds that have been approved for clinical trials

SECTION 6

HIGH-THROUGHPUT ADME COMPOUND PROFILING OFFERING In the pharmaceutical industry, early liability assessment of drug candidates is an important element to decrease the high attrition rate in the drug discovery and development process. One of the key challenges is the balance between drug efficacy and potential adverse effects at the earliest possible point in time for de-risking cost-intensive activities especially during late-stage clinical development.

Joachim Kraemer Senior Vice President Bioreagents, Biophysics and Assay Development

Portfolio of high-throughput ADME Profiling assays

drugs can be identified activating the human pregnane X receptor (“PXR”) to induce CYP450 3A4. Additional critical ADME assays identify inhibitors of the hERG, Nav1.5 or Cav1.2 ion channels which are implicated in adverse cardiovascular events.

ADME assays for four highly relevant CYP450 isoforms are established on our high-throughput RapidFire/MS platform. Medium-throughput LC/ MS/MS based assays are provided To meet this demand Evotec has for CYP450 1A2 and 2C19. established an industrialised highthroughput ADME profiling service In support of discovery chemisfor routine compound screening try programmes, externalization of against a panel of the most criti- safety pharmacology profiling is cal safety pharmacology targets. providing a unique business opporPotential drug-drug interaction can tunity for our partners to reserve be determined by using our panel internal capacity for core research of cytochrome P450 (“CYP450”) activities, expand their ADME reversible inhibition (“RI”) assays assay portfolio and to mitigate risk. for the six main CYP450 isoforms in Besides these benefits high-throughcombination with time-dependent put ADME profiling is a time- and inhibition (“TDI”). Furthermore, cost-efficient process consistently

ADME ASSAY

READ-OUT

THROUGHPUT COMPOUNDS/WEEK

CYP450 3A4, 2C8, 2C9, 2D6 (RI)

RapidFire/MS

>800

CYP450 2C8 (RI)

RapidFire/MS

>300

CYP450 1A2, 2C19 (RI)

LC/MS/MS

>150

CYP450 3A4 (TDI)

RapidFire/MS

>600

CYP450 3A4 (induction of PXR receptor)

Luminescence

>600

hERG

Radioligand binding

>800

hERG

Ion Works® Quattro™

>100

Nav1.5, Cav1.2

FLIPR™

>800

25

delivering highest data quality.

Pharmacological sensitivity and statistical robustness of the ADME assays is routinely monitored by a range of reference compounds

ADME assays are performed according to validated SOPs and provide 8-point IC50/EC50 for test compounds in triplicate Turnaround time from compound reception to data reporting is 7 working days Assays are scalable towards higher throughput

platforms. Ion channel electrophysiology assays run on Ion Works® Quattro™ automated patch clamp platform

State-of-the-art read-out technologies including RapidFire/MS, FLIPR, SPR, fluorescence, radiometric, high-content imaging Flexible data analysis with internal or external software tools enabling rapid data uploads Professional project management with experienced leads KINAFFINITY® COMPOUND PROFILING Numerous small molecule kinase inhibitors are currently being developed for treatment of cancer, inflammation or autoimmune diseases.

However, drug discovery faces significant challenges, particularly due to unacceptable safety pharmacology profiles caused by an inhibitor’s lack of selectivity. Undesirable off-target kinase profiles frequently induce toxicity that might halt the development of candidate drugs. To decrease the high attrition rate the design of selective as well as multispecific inhibitors is becoming increasingly important.

TECHNOLOGY PLATFORM AND EXPERT KNOWLEDGE Within many collaborations with major pharmaceutical and biotech companies, Evotec has successfully provided expert knowledge in development and integration of complex processes for our collaborator’s drug discovery programmes including: Logistics of compound shipments in close interaction with global carriers

Management of large libraries and regularly shipped smaller compound batches for ADME or SAR profiling Broad expertise in assay development with > 400 biochemical, biophysical and cell-based assays Compound profiling using bench- KinAffinity® is Evotec’s hit-to-lead top and fully automated screening compatible approach to profile drug

candidates against endogenously expressed, full-length proteins in the presence of cellular co-factors and native complex partners – a major advantage over traditional biochemical panel screening using only recombinantly expressed, purified proteins or protein domains. Evotec’s platform: Enables rapid target profiling of kinase inhibitors in cell and tissue samples. Unlike traditional biochemical kinase panel screening, the inhibitor’s target affinities are determined simultaneously for a large number of native kinases within their physiological cellular environment Identifies target interactions possibly requiring additional co-factors or the formation of multi-component complexes, thereby complementing traditional biochemical assays that use only recombinant proteins Can determine a target’s Kd value to a free compound without needing to immobilise the inhibitor Employs a ready-to-use affinity matrix comprising well-characterised broad-spectrum kinase inhibitors to enrich the subproteome of endogenously expressed kinases of cells or tissues

Test compound

1 Cell or tissue sample

2 Mixture of beads of broadly selective kinase inhibitors for enrichment of kinases

3 Competition of target kinases by test compound

4 LC-MS analysis

(1) A lysate of the cell line or tissue of interest is prepared. (2) A matrix comprising of immobilized broad-spectrum kinase inhibitors is used to enrich the expressed kinome of the cell or tissue sample. (3) Competition assays with the free test compound and (4) quantitative mass spectrometry analysis reveal the compound’s affinity profile, (5) ranking all protein kinase targets according to their Kd values for the free compound.

5 List of target kinases ranked according to their K d values Sepharose resin Immobilised kinase inhibitor test cpd Target kinase Target kinase bound to test cpd

SECTION 7

PROTEOMICS OFFERING Evotec offers unique proteomics services to address key issues in drug and biomarker discovery. We continuously advance our capabilities in mass spectrometry-based proteomics to ensure unrivalled comprehensiveness and data quality when analyzing cells, animal models and patient samples.

This involves: Highly optimised experimental strategies tailored to different proteomics applications and project needs Industry-leading capabilities in high-end quantitative mass spectrometry Experience and infrastructure to analyse the enormous amounts of data generated in large-scale

Henrik Daub Senior Vice President Chemical Biology

TARGET DECONVOLUTION

TARGET DISCOVERY

Target ID & validation

proteomics projects

Advanced statistics and bioinformatics for systems-wide data analysis In-depth data interpretation to extract relevant drug target and mode-of-action information from proteomics experiments Extensive track record to deliver high-quality results within agreed timelines Evotec’s chemical proteomics methods such as Evotec Cellular Target Profiling™ reveal cellular drug targets and selectivity in an unbiased manner. Other Evotec proteomics technologies record protein modification and expression on a proteome-wide scale for drug mode-of-action analysis or biomarker discovery efforts. Thus,

CELLULAR MODE-OF-ACTION ANALYSIS TO SUPPORT LO

HT SCREENS/ SCAFFOLD SELECTION

Screening

Evotec Cellular Target Profiling™ Subproteomic Evotec Cellular Target Profiling™

ON/OFF TARGET PROFILING

H2L

EVOTEC CELLULAR TARGET PROFILING™ TO SUPPORT H2L

Omics Technologies (Phosphoproteomics, Acetylomics, Proteomics)

MODE OF ACTION ANALYSIS

LO

DRUG REPOSITIONING

RESPONSE PREDICTION

Pre-clinic

BIOMARKER ASSAYS

PhI/II

PROTEOMIC SIGNATURES FOR BIOMARKER DEVELOPMENT

27

our platform offers highly innovative solutions that can be leveraged across the entire drug discovery and development pipeline.

QUANTITATIVE CHEMICAL PROTEOMICS Evotec scientists have pioneered chemical proteomics applications for the identification of cellular small molecule targets. The basic concept involves functional immobilisation of an appropriate linker derivative of a drug, followed by affinity purification of target proteins from cell or tissue extracts and finally their identification by MS analysis. Unlike other competitor technologies, our Evotec Cellular Target Profiling™ technology captures target specificity and affinity information by

quantifying target binding across successful profiling of diverse defined sets of affinity purificasmall molecule compounds tion and drug competition experiments.

Selectivity data about cellular onand off-target liabilities is particuEvotec Cellular Target Profiling™ larly useful to inform decisions at

Reveals and verifies specific various stages of drug development, cellular targets of a drug by for example in the lead optimisation quantitative mass spectrometry phase or in the pre-clinic candidate Determines target-specific selection process. Moreover, chemidissociation constants for the cal proteomics enables target deconcompound studied, ranking volution of bioactive compounds targets according to their likely identified in phenotypic screens. physiological relevance Cellular target identification is the Performs selectivity analysis on crucial step to enable further drug a proteome-wide scale against optimisation and development. native, post-translationally Evotec Cellular Target Profiling™ modified proteins in the pre- perfectly complements Evotec’s sence of cellular co-factors and medicinal chemistry and highcomplex partners content screening capabilities to Has an extensive, non-target deliver new drugs and targets across class restricted track record in many therapeutic areas.

Evotec Cellular Target Profiling™ key elements Test compound

1 Cell or tissue sample

2 Enrichment of target proteins by compound affinity chromatography

3 Competition of target proteins by test compound

4 LC-MS analysis

(1) A lysate of the cell line or tissue of interest is prepared. (2) A linker derivative of the test compound is immobilised on sepharose beads and used to enrich compound target proteins. (3) Competition assays with the free test compound and (4) quantitative mass spectrometry analysis reveal the compound’s affinity profile, (5) ranking all protein targets according to their Kd values for free compound.

5 List of target proteins ranked according to their K d values

Sepharose resin Linker derivative of test cpd test cpd Target proteins

SECTION 7

PhosphoScout® Workflow

Proteins

Peptides

Global phosphoproteome enrichment

LC-MS

Isotope labelling

GLOBAL ANALYSIS OF PROTEIN MODIFICATIONS Evotec’s proteomics platform for global protein modification analysis permits the identification and quantification of more than 10,000 phosphorylation sites, 1,000 lysine acetylation sites or 5,000 ubiquitinylation sites in a single experiment, which may be performed in cells, tissues or patient samples. Accurate quantification is enabled by differential isotope labelling, while highly optimised peptide enrichment and analysis on fast, sensitive and accurate mass spectrometers ensure highest coverage. Evotec’s platform provides highly reliable results, both for in-depth bioinformatics involving enrichment, cluster, network and motif analyses, as well as for expert data interpretation on the level of site-specific changes.

unbiased and comprehensive investigation of signalling pathways, to delineate the cellular modes of action for kinase inhibitors. Typical applications include: Identification of specific pharmacodynamic read-outs for therapeutic kinase inhibition Selection of lead compounds or pre-clinical candidates with maximal on- and minimal off-target activity in cellular conditions, either as single agents or in combination with other drugs Monitoring phosphoproteome regulation in different biological models to shed light on factors underlying differential biological activity of lead compounds or preclinical candidates

In addition to PhosphoScout®, Evotec’s acetylomics and ubiquitinomics platforms offer similar appliOur quantitative phosphoproteom- cations for HDAC and E3 ligase ics (PhosphoScout®) enables the inhibitors using highly optimised

workflows with regard to sensitivity and selectivity. As a result, these technology platforms enable comprehensive mode of action studies in the biologically relevant context.

GLOBAL PROTEOME PROFILING AND BIOMARKER DISCOVERY Evotec offers industry-leading platforms for comprehensive protein expression profiling allowing unbiased, proteome-wide target and biomarker discovery on the functional protein level. Depending on the application and required throughput, tailored proteome analysis workflows are available:

Deep proteome profiling for de­tection of up to 10,000 proteins from cell or tissue samples upon efficient peptide pre-fractionation Single-shot proteome profiling to a depth of 5,000 proteins in cell or

29

Data analysis

tissue lysate, enabling comparative proteome analysis across 100+ samples

Comprehensive analysis of any type of biological material, including formalin-fixed paraffin-embedded (“FFPE”) solid tumor samples and body fluids such as plasma and CSF Targeted mass spectromety analysis by Multiple Reaction Monitoring (“MRM”) fully established at Evotec for follow-up validation experiments

Both the expertise and the data processing infrastructure are in place for accurate quantification across many different samples, such as whole panels of cell lines, xenograft models or patient tissue samples. Such data, together with drug response information, provides a unique basis for the discovery of predictive protein biomarkers, arguably one of the most exciting proteomics applications in future medicine.

1

Pre-clinical biomarker discovery in cultured or primary cells

Early identification of predictive biomarker candidates by proteome profiling of cell line panels

2

Pre-clinical biomarker discovery in animal models

Unbiased biomarker identification from mouse xenograft models by global and targeted proteome analysis

3

Clinical biomarker in patient samples

Proteome-wide biomarker identification from patient-derived leukemia cells or FFPE tumor tissue

EVOTEC’S THREE-TIER STRATEGY TO SUPPORT ONCOLOGY BIOMARKER DISCOVERY Evotec technologies cover the entire biomarker discovery phase including elaborate bioinformatics for the identification and verification of predictive multivariate markers, so-called protein signatures. These capabilities have been demonstrated in various cancer-related projects, delivering response prediction signatures in lung cancer cell lines, mouse xenograft models and leukemia patients.

In conclusion, Evotec offers an integrated and unique platform of innovative proteomics technologies for drug and biomarker discovery. Moreover, Evotec is constantly investing in R&D activities dedicated to advance its high-end proteomics capabilities and maintain industry leadership in the years to come.

SECTION 8

CELL & PROTEIN PRODUCTION

There are numerous reasons why a target may not be stably expressed in mammalian cells and a transient approach may be more appropriate for cell-based screening. To address this problem, Evotec routinely uses a variety of transfection technoloEvotec’s wealth of expertise covers gies from lipid based to viral mediall principles including cell line ated delivery to achieve transient generation, small- or large-scale expression. transient transfection, frozen cell technologies and protein expression Particularly the MaxCyte electropofrom small-scale proof-of-concept ration platform offers access to large studies through to batch production batches of up to 10 billion cells, a for HTS, counter screening activities cost-effective and efficient transas well as structural biology applica- fection process. Transiently transtions. For clients looking to outsource fected or virus transduced cells can the management and supply of their be provided as assay-ready frozen own cell collections, Evotec offers instant cells, which are functionally a cell bank management scheme, expressing the target instantly after establishing, propagating and distrib- thawing. uting thousands of cell lines between various locations.

The availability of cells is a critical bottleneck in screening campaigns.

CELL LINE GENERATION Frozen instant cells, which can be AND BANKING assayed without prior cultivation From recombinant cells which have have become a valid and frequently been either generated by stable used alternative to cells from a transfection or viral transduction, continuous growing culture. monoclonal cell lines are isolated and screened for their functional Evotec’s cell culture unit was one of expression of the target protein. the first suppliers of frozen instant After a first broad screening on cells for global pharmaceutical expression level using immuno­ research. Cells are: Expanded under controlled fluorescence labelling, Westernblot, culture conditions to large bulk or RT-PCR, usually a secondary quantities screening of selected clones by a

Harvested applying a gentle cell-based assay is applied. functional assays, assay development, HTS and compound profiling offers clients the best-possible platform for cost-efficient and timely delivery of highest quality data for their drug discovery programmes.

Mark Slack Vice President In vitro Pharmacology

OFFERING Evotec provides access to a comprehensive base of services and capabilities around cell and protein production. These can be accessed as a stand-alone function or as an integral part of a wider collaborative programme accessing our drug discovery expertise.

Combining expertise in cell culture and protein production methods with expertise in

31

Cell line generation (n=59 projects)

GPCR 23

Others 9 Ion channel 7 Reporter cell lines 3 Enzymes 3

method which does not affect surface receptors Prepared to a density which corresponds to the requirements of your assay, the cells are dispensed into vials using an automatic filling device. This fast processing of the cells significantly decreases the time from harvest to freezing, which is critical for maintaining the quality of frozen instant cells Frozen in a controlled rate freezer and stored at -150°C until they are shipped to you For secondary screening or drug profiling, in which you need only a few assay plates per day over a longer period of time, we can apply a proprietary technology to freeze cells directly in assay plates which can then be used upon demand. For ligand binding assays, we offer cell membranes in bulk.

All batches of cells and cell-based products are carefully controlled for their quality before they are shipped to you. Frozen instant cells are checked not only for viability but also for their ability to adhere shortly after thawing. A good „fitness“ of the cells is often correlated with fully functional response.

Finally, the assay performance of the frozen cells is compared to cells from a continuous growing culture considering Z’, S/B and EC50.

Kinases 4

Transporters 4 NHRs 1 Cytokine receptors 5

Transient transfections (n=56 projects)

GPCR 13 Others 17

Enzymes 2 Proteases 13

Ion channel 9 Transporters 2

Membranes (n=40 projects) Others 5

Transporters 6

GPCR 19

Ion channel 10

Assay ready frozen cells (n=223 projects)

GPCR 96 Others 35

Ion channel 46 Primary cells 10 Kinases 8 NHRs 10

Transporters 18

SECTION 8

DNA engineering

Stably transfected cells

Transiently transfected cells

Non transfected cells

Bulk cell production

Frozen instant cells

Membrane preparation

Custom cell & reagent supply

Evotec offers professional management and maintenance of client cell banks encompassing the addition of new cell lines and shipment of assay ready cells and vials to the clientâ&#x20AC;&#x2122;s research centres. A dedicated team with access to the necessary bioinformatics, quality control processes and infrastructure can additionally provide support in the sourcing and maintenance of disease relevant cell banks such as oncology. The databases covering all aspects of the respective cell line such as growth rates, morphology and genetic confirmation are available over a web-based client portal providing clients with the essential access to all necessary information.

Recombinant proteins

Evotecs screening services

RECOMBINANT PROTEIN PRODUCTION Protein production lies at the heart of drug discovery. Evotecâ&#x20AC;&#x2122;s recombinant protein production facilities provide comprehensive coverage of expression hosts, purification techniques and analysis tools and delivers a unique solution for the efficient Construct design delivery of highquality proteins in a timely fashion. Our capabilities can be accessed flexibly, Quality either as standalone units or as part of an integrated drug discovery programme.

All proteins are prepared with end use in mind, including the delivery of high-quality proteins for bioassay and biophysical applications

including the labelling of proteins for NMR and mass spectroscopy, material for SPR and ITC and crystal grade material for macromolecular X-ray crystallography.

The production platform delivers on hundreds of recombinant proteins for drug discovery per year to our clients.

Cloning & expression

Established workflows for all major activities involved in protein production lead to routine and Protein reproducible results. purification Our highly experienced team of scientists will optimise expression levels and purity using a suite of techniques, including semi-automated expression analysis and purification, to guarantee quality in a realistic timeframe.

33

E. coli up to 120L/week

Tags: no tag, His (N- or C- terminal), GST, MBP, Trx Strains: BL21 (DE3)/pLysS, ArcticExpress, BL21-AI Secretion signal peptide (if needed): pelB

Pichia pastoris up to 20L/week

Tags: no tag or C- terminal His-tag Strain: X-33 Secretion signal: α-Factor

Insect cells (Baculovirus) up to 80L/week Mammalian cells up to 30L/ week

Expression hosts in 2013, >400 production runs completed

E. coli 48%

Insect cells 37%

Pichia pastoris Mammalian 1% 14%

Uses of protein

Crystallo 58%

Assays 37%

Other NMR 2% 3%

Tags: no tag, His (N- or C- terminal), GST, MBP Strains: Sf21, Sf9, T. ni Secretion signals (if needed): gp67, honeybee melittin, wild type Tags: no tag, His (N- or C- terminal), GST, MBP Strains: CHO, HEK293, CAP-T Secretion signals (if needed): wild type, lgk

Specific expertise in the team covers the common protein families and also extends into the more demanding areas of macromolecular complexes (protein:protein and protein:DNA) and membrane proteins.

All main expression hosts are supported by Evotec with shuttle vectors available to rapidly move expression to higher organisms as required. Gene synthesis is performed by a series of secure third party relationships with confirmed timelines.

The team has unparalleled experience in labelling of proteins for NMR work including ubiquitous 15 N, 13C and 2H in E. coli and yeast and side chain specific labelling in insect expression hosts.

In addition to purification of proteins

by SOPs or according to literature precedents, Evotec has extended methods commonly employed in structural genomic facilities so that the team can perform Highthroughput (“HTP”) expression screening in E. coli, insect and mammalian hosts to determine optimal conditions for production ahead of large scale production runs. Large scale expressions can be carried out in parallel, either in shake flasks or in eight WAVE cellbag bioreactors, each capable of producing up to 25-L of material for purification.

SECTION 8

Option 1 – 24/week

IMAC His-Trap (Gravity)

Option 2 – 16/week

Option 3 – 8/week

IMAC His-Trap (Akta Xpress)

IMAC His-Trap (Akta Xpress)

Insufficient yield

Insufficient yield SEC

QC

m/z

m/z

SDS-PAGE, Western blotting, M.S.

Further scale-up/ optimisation e.g. 10-20L, as desired

No

Yield and quality OK?

Insufficient yield

Yes

Dispatch

Three example methods for HTP purification are shown. Either single step IMAC columns run by gravity (Method 1) or on an Akta Xpress (Method 2) or a two-step process on Akta Xpresses (Method 3) may be performed

With fifteen Akta purification modules and HTP methods suitable for cost-effective parallelization, Evotec has the capacity to deliver significant quantities of proteins in realistic cycle times through a variety of purification schemes (see figure above).

All proteins pass stringent in process quality controls, selected from a panel of options including Mass Spectroscopy, Dynamic Light Scattering, Differential

Scanning Fluorimetry, Activity Testing and NMR, before final release to the end user. In summary, by tailoring the purification schemes to the problem to be solved and using appropriate input from scientists on the projects, Evotec delivers the target protein optimised for the end use.

Driven by demand the reagent production team will be expanding in 2015. At the end of March 2015 a new facility in Princeton, US will be operational across cell science and recombinant protein production. This facility will deliver the range of high quality services currently offered from our European sites, but will be able to provide the fastest possi­ ble cycle times for business critical deliveries.

35

LABELLING CHEMISTRY Evotec offers labelling of biological relevant molecules and reagents for use in ELISA, FACS, FCS, plate readers and other fluorescence based detection systems. Our service helps clients to improve their research results in all areas of biological and pharmacological assays, as well as clinical diagnostics and analytics. Our expertise covers the labelling of the following: Small organic molecules e.g. hormones, neurotransmitters Peptides and chemokines (as enzyme substrates, GPCR ligands, etc.)

Proteins including antibodies Site-specific mutagenesis of proteins (for site-specific labelling) Glycoproteins

Carbohydrates Lipids (for membrane studies) Beads and surfaces Evotecâ&#x20AC;&#x2122;s labelling chemistry team has acquired a comprehensive expertise having successfully designed and labelled components for more than 150 different assays representing a broad range of assay classes. For labelling we use the optimally suited dyes and tags for your application, such as: Proprietary dyes (EVOblueâ&#x201E;˘ family) Commercially available fluorescent dyes and the in-licensed MR121 dye Non-fluorescent markers and probes like affinity tags, biotin, reporter enzymes, etc.

A2a receptor purification using cobalt IMAC at Evotec (left and center) yields sufficient protein for crystallisation by the lipidic cubic phases (LCP) technique

Crystals

SDS-PAGE Co2+ IMAC

Western Blot Co2+ IMAC

SECTION 9

IN VITRO

PHARMACOLOGY OFFERING The cornerstone of Evotec’s in vitro pharmacology function is disease and target biology expertise coupled with state-of-the-art technology platforms. A large team of scientists with extensive industrial experience supports the in vitro pharmacological characterisation of compounds as part of hit expansion, lead finding or lead optimisation projects. Our team routinely generates projectrelevant high-quality data in short turnaround times.

Andreas Scheel Executive Vice President In vitro Biology In vitro Pharmacology

Typical activities include: Development of biochemical and functional assays Secondary and tertiary characterisation of screening hits Compound characterisation as part of hit-to-lead and lead optimisation programmes: — Design and implementation of target-relevant assays (screening cascades) — Potency and selectivity testing — Mode of action studies (e.g. competitive versus allosteric mechanisms, reversibility, usedependent mechanisms for ion channel modulators) — Translational assays: testing of compound potency and mechanism using disease-relevant primary cells from rodents, primates or human

In more than 15 years of compound profiling at Evotec, >400 assays have been developed and executed.

Such activities are part of Evotec’s integrated lead finding and optimisation projects but are also frequently used to support medicinal chemistry projects that are executed in the labs of Evotec’s partners.

AVAILABLE READ-OUT AND ASSAY TECHNOLOGIES Evotec has access to a wealth of assay technologies that can be utilised to assess compound activity. Appropriate technologies and expert teams are selected to answer key project questions and to ensure project advancement. Beyond standard and established read-out technologies for biochemical and cellular assays, Evotec has built key expertise in a number of technological areas that have shown to

Studying compounds modulating beta cell proliferation

Insulin

BrdU

37

be drivers for the success of our partner’s projects.

These include: The use of stem cells to derive neurons and primary neuronal cultures to build disease-relevant cellular models and to identify and characterise new compounds with disease-modifying properties

An extensive knowledge of highcontent screening and a state-ofthe-art hardware platform to run complex and disease-relevant imaging assays, e.g. using primary neuronal cultures, kidney cells as well as rodent and human beta cells A world-leading ion channel discovery platform including fluo-

rescence-based assays, automated and manual patch clamp methods An excellent suite of bio­physical methods including SPR and NMR that are utilised as part of our structure-based drug design projects but also LC-MS-based methods that are utilized to assess difficult-to-assay enzyme targets

Neuroprotective compound

Read-out technologies CELL-BASED ASSAY TECHNOLOGIES Fluorescence read-outs —H TRF, standard dyes, ligand binding Second messengers (Ca2+, cAMP, IP3) Membrane potential (GPCRs, ion channels and transporters) Reporter gene assays ELISA (standard and Mesocale) Imaging (HCS) — OPERA® and Zeiss Radioactive binding and uptake Flow cytometry Label-free Migration assays Whole cell blood assays Metabolic analysis with Seahorse using Primary cell culture Stem cells Established cell lines

BIOCHEMICAL ASSAY TECHNOLOGIES FCS+plus Fluorescence polarisation Fluorescence intensity HTRF/Delfia AlphaScreen Luminescence LC/MS BIOPHYSICAL READ-OUT TECHNOLOGIES Surface Plasmon Resonance/SPR Mass Spectrometry/LC/MS Nuclear Magnetic Resonance/NMR Radiometric Thermal Shift ELISA

SECTION 9

OVERVIEW TARGET CLASSES Our in-depth experience in the biology and pharmacology of diseaserelevant target classes is what our partners come to us for. This expertise is a key driver to the successful and rapid execution of lead finding and optimisation processes. Evotec’s in vitro Pharmacology team has gained expertise across a wide area of disease targets. Beyond a large number of projects that have successfully been run in the classical target areas such as GPCRs, ion channel and kinases, we have also worked on a large diversity of other target areas such as transporters, proteinprotein interactions and multiple enzyme families, including novel target classes such as epigenetic regulators.

Identifying cpds that protect podocytes in chronic kidney disease

tion, pain, inflammation, metabolic disease, oncology and immunology.

Our scientific expertise and understanding of disease mechanisms DISEASE EXPERTISE AND combined with our track record in TRANSLATIONAL ASSAYS setting up relevant in vitro models Evotec’s core expertise includes is a key factor in our success when areas such as CNS, neurodegenera- working with our partners.

Co-culture system of stem cell derived motoneurons and astrocytes together with microglia to identify cpds for the treatment ALS

Neurite Total Length

100K 80K 60K 40K 20K

2

3

4

5

Days After Plating

6

7 D3 After Plating

GFAP/GFP/DNA

Setting up complex in vitro assays utilising rodent or human primary cells to confirm the potency and mechanism of lead compounds is a prerequisite to build confidence in the translatability of any mechanism. We build these assays early in the drug discovery process to gather disease-relevant data before assessing compound efficacy in vivo. Such assays are also utilised to identify read-outs for in vivo target engagement. We routinely utilize various neuronal and stem cell cultures, pancreas and beta cells, kidney cells and various immune cells to assess bespoke read-outs and mechanisms, including cell health and survival, apoptosis, differentiation, de- / regeneration; neurite outgrowth, retraction and synaptic density; cellular signalling and secretion.

39

Extensive expertise with translational assays using primary cells and tissues to investigate compound potency and mechanism

Rodent glomeruli

Synaptic connectivity

Human kidney cell fibrosis

Replicating pig beta cells

ISOLATION, CULTIVATION AND MANIPULATION OF PRIMARY CELLS Neurobiology: neurons (CNS, DRGs, moto足 neurons), astrocytes, microglia including co-cultures Blood: PBMC, TH1/TH2 populations, B cells Pancreas: islets, beta cells Kidney: podocytes, glomeruli

ASSAY READ-OUTS Cell density, degeneration, regeneration, survival Cytokine secretion Transcrptional activity Protein phosphorylation ASSAY TECHNOLOGIES Imaging MSD LC-MS Standard methods

SECTION 10

ION CHANNELS OFFERING Evotec’s world-class ion channel platform combines industry-driven expertise with state-of-the-art instrumentation. In routine use are multiple setups for manual patch clamp studies and all of the most popular automated patch clamp instruments.

The automated patch clamps or FLIPRs can be used as entry points for HTS, and the manual and automated patch clamps enable on-going support of medicinal chemistry programmes progressing from HTS through hit expansion, hit-to-lead, lead optimisation but are also utilised for safety pharmacology.

Stephen Hess Research Leader Ion Channels

OVERVIEW OF ION CHANNEL SCREENING AT EVOTEC Evotec has a strong background in running hit identification campaigns against ion channel targets.

On average more than 235,000 compounds per target are screened in singlicate at a fixed compound concentration. Campaigns resulted in “primary” hit rates in the range of 1% and confirmation rates are generally good (>50%), underlining robust assay performance. Secondary assays using electrophysiology to confirm compound activity is part of the routine HTS follow-up and are key to the successful identification of relevant starting points for medicinal chemistry projects.

The electrophysiology platforms in place at Evotec

IonWorks® Quattro™

Qpatch HTX®

PatchLiner®

Dynaflow® Pro II

Manual Patch-Clamp

PLATFORM IonWorks® Quattro™

2

384-well

4,000-10,000

Q-patch HTX

2

48-well

150-450

Hit-to-lead, lead optimisation and early safety assessment

PatchLiner©

3

16-well

75-150

Hit-to-lead, lead optimisation and early safety assessment

Port-A-Patch®

2

1-well

25-50

Manual Electrophysiology (Dynaflow™ optional)

5

1-well

1-25

®

High-throughput screening, hit-to-lead

Hit-to-Lead, Lead optimisation Lead optimisation, safety pharmacology

41

TRACK RECORD OF SUCCESS WITH A RANGE OF ION CHANNEL CLASSES Evotec has electrophysiology and ion channel pharmacology expertise using transiently- &

1

Voltage gated ion channels Potassium ion channels: Kv family Calcium channels: Cav family Sodium channels: Nav family

2

Additional potassium ion channels KCa (e.g. SK, BK, IK) Inward rectifiers (e.g. Kir2.1) Tandem pore domain (e.g. TREK, TASK)

3

Ligand-gated ion channels P2X family (e.g. P2X3, P2X7) Glutamte receptors (e.g. NMDA, AMPA) TRP family (e.g. TRPV1, V3, V4) GABA-A

4

Others CRAC Chloride channel (e.g. CFTR)

stably-expressing cell lines or cells from primary tissues (e.g. DRGs, NDGs). In addition, we have capabilities and experience in the generation of transient and stable formats for ion channel targets.

EXPERTISE IN IDENTIFYING CHANNEL OPENERS AND BLOCKERS

DEMONSTRATED CAPABILITY IN ION CHANNEL DRUG DISCOVERY ACROSS VARIOUS PLATFORMS FROM 1 TO 384 WELL FORMAT

NDG neurons 6h after preparation

NaV current

Electrophysiological recordings from primary cells provide additional mechanistic insight beyond that possible using recombinant cells. In this case we used neurons isolated from dorsal root and nodose ganglia. On the right is an example taken from our efforts using mouse nodose ganglion neurons:

TTX 300nM

NaV channel X: 1.00 ms

Y: 2.00 nA

Voltage-gated sodium channels (NaV) are expressed in neurons The selective NaV blocker, TTX (300nM) inhibited NaV signal in big diameter cells (arrows)

SECTION 11

IN VIVO

PHARMACOLOGY/DMPK

Roland Wehr Senior Vice President In vivo Pharmacology

OFFERING Animal models remain crucial for providing a holistic understanding of how drugs work in vivo. Evotec offers in vivo pharmacology services following custom-designed protocols to support your drug discovery needs from delivering single studies to taking over responsibility for planning the in vivo pharmacology strategy of integrated drug discovery programmes. Evotec’s in vivo pharmacology team consists of highly experienced scientists with extensive pharmaceutical and biotechnology backgrounds. Evotec has state-ofthe-art animal facilities equipped to house immune-compromised animals, genetic models and other rodents. All experimental procedures involving animals are approved according to EU and federal law.

Evotec develops bespoke assays to support drug discovery programmes to drive Structure–Activity Relationship (“SAR”) and development of PK/PD relationships. We further offer high-quality in vivo services including target validation, tolerability studies, efficacy screening, in vivo pharmacology profiling and mode-of-action (MOA) studies in a wide range of therapeutic areas.

METABOLIC DISEASE Models include type 2 diabetes, obesity and beta-cell regeneration Models: fa/fa- and ZDF-rats, db/ db-, ob/ob- mice; Diet-Induced Obesity (“DIO”) mice; Ins2-Akita mice; Chemically-induced diabetes using STZ/Alloxan in rodents (neonatal and adult) Read-outs Body weight, food intake and water consumption Blood glucose, glycated hemoglobin HbA1c; clinical blood chemistry Body composition analysis (NMR) Glucose & insulin tolerance test Hyperinsulinemic euglycemic clamp Locomotor activity & body core temperature Plasma and tissue analysis (e.g.

ELISA, qRT-PCR) Energy expenditure (indirect calometry; O2 /CO2 analysis) Morphometric analysis of the pancreas and adipose tissue Beta-cell regeneration Blood pressure analysis

43

DIABETIC COMPLICATIONS & KIDNEY DISEASE

Models of diabetic complications (retino- & nephropathy): STZ induced diabetes, Oxygen-Induced Retinopathy (“OIR”) model; ZDFrats, db/db-, ob/ob-, Ins2-Akita mice.

Models of acute and chronic kidney disease: Surgery models (IRI: Ischemia/Reperfusion Injury, UUO: Unilateral Ureteral Obstruction); adenine induced chronic kidney failure.

Read-outs Urin analysis: albumin and creatinine ratio; kidney disease biomarker Plasma/serum: Glomerular Filtration Rate (“GFR”); both creatinine and FITC-inulin based; hormones i. e. insulin, leptin, cytokine profiling, clinical blood chemistry, glycated hemoglobin HbA1c Immunohistochemistry & histo­ logy ie. HE, PAS, Masson’s Trichrome, Sirius Red

Diabetic effect of Pioglitazone in diabetic male ZDF rats Vehicle

10

Pioglitazone 10 mg/kg*day; p.p.

HbA1c in %

9

8

7

6

5 0

5

10

15 days treatment

20

25

INFLAMMATORY DISEASE

Models: Collagen AntibodyInduced Arthritis (“CAIA”), Peptidoglycan-Polysaccharid (PG-PS) induced arthritis, mustard oilinduced neurogenic inflammation Read-outs Visual scoring of arthritis, plethysmography, caliper measurements Cytokine analysis and clinical blood chemistry Histology (histopathological scoring; immune-histochemistry) Immunohistochemistry & histology ie. HE, PAS, Masson’s Trichrome, Sirius Red

SECTION 11

Efficacy of Gabapentin in a neuropathic pain model (SNL=Spinal Nerve Ligation)

50% response threshold (g)

15

10

*

* 100mg/kg p.o.

*

*

30mg/kg p.o.

10mg/kg p.o.

5 Vehicle p.o.

0 pre-SNL

post-SNL

0

60

120

240

Time post treatment (min)

ACUTE AND CHRONIC PAIN Models: Neuropathic pain: Spinal Nerve Ligation model (“SNL”); Visceral pain: colorectal/vaginal distension-induced visceromotor reflex; Inflammatory pain: CFAinduced mechanical and thermal hyperalgesia, formalin test.

Read-outs Thermal sensitivity: Hargreaves

Test, hot plate, tail flick Mechanical sensitivity: Von Frey

test, pressure application measurement device Flinching behaviour (automated nociception analyser) Visceral motor reflex-EMG activity

NEURODEGENERATIVE ONCOLOGY DISEASE Models: Subcutaneous human Models: transgenic mice and rats xenograft models for Chorea Huntington and mice Read-outs for Alzheimer’s disease.

Survival, clinical signs, Read-outs body weight Locomotor activity (rotarod), Tumor volume, T/C emotion (Elevated Zero Maze, As part of Evotec’s integrated drug fear conditioning), cognition discovery platform, the in vivo (Novel Object Recognition, pharmacology is further supported spatial memory), Irwin test Pre-pulse inhibition, gait analysis by DMPK and histology.

45

Oral bioavailability study in rats

Plasma Concentration (nM)

1000

100

intravenous oral

10

1 0

4

8

12

16

20

24

Time (h)

PHARMACOKINETICS Obtaining pharmacokinetic data is a key requirement in the evaluation of new chemical entities. Evotec offers: Various administration routes: intravenous (incl. infusion), per os, intraperitoneal, subcutaneous, intra-cerebrospinal and intra­ muscular Cassette PK screening of up to 5 compounds simultaneously

Different sampling routes: jugular vein cannulation, cardiac puncture, tail vein microsampling, retro-orbital Different matrices: blood, plasma, cerebrospinal fluid, tissues, bile, urine and faeces Data generated with the latest version of Phoenix® WinNonlin®6.3 software, analysis performed using rigorous accep­tance criteria

WHY EVOTEC? As a core value in drug discovery Evotec offers a wide range of validated and routinely used in vivo pharmacology models, DMPK as well as histology services. Evotec works closely with our clients to ensure that any custom protocols are executed to their exact specifi­ cations, adapting the latest procedures from the literature. Finally Evotec will provide a detailed and accurate report on your study.

Immunohistochemistry and morphometric analysis of male ZDF rat pancreas: Islet of Langerhans fragmentation in male ZDF rats

HISTOLOGY, IMMUNOHISTOCHEMISTRY AND MORPHOMETRIC ANALYSIS The combination of in vivo pharmacology with histology techniques offers a powerful tool to further analyse disease relevant biomarkers.

Whereas histology and IHC helps to identify disease and target relevant changes, automated morphometric analysis can yield unbiased quantitative information amenable to statistical analysis, which adds significant value to the customers drug discovery effort. Furthermore, histopathology on critical organ systems will allow early risk assessment.

SECTION 12

EVOMAB

As a partner in antibody drug development, Evotec offers cell lines over-expressing a given target or target domains for primary selections using the customers own antibody libraries and platforms.

Arnd Steuernagel Senior Vice President Biologics

OFFERING Evotec offers its HTS/HCS platform capabilities for antibody identification including assay development, screening, hit characterisation up to in vivo pharmacology.

screening. These processes bias the screening pool in favour of affinity binders with the risk missing potentially interesting functional antibody clones. EVOmAb addresses this problem by offering early entry into functional selection using Evotec’s high-throughput antibody expression and screening capabilities.

The antibody expression platform combines high throughput cloning with high parallel expression and purification strategies. This platform enables the full utilisation of Evotec’s established high throughput screening capabilities.

The offering includes reformatting antibody pools from phage display selections or animal immunizations into suitable screening formats. We provide optimal cell-based HTS/ HCS screening formats supporting fast development cycles.

Evotec offers a turnkey solution for antibody development up to the production of mg quantities necessary for testing in animal models.

Evotec has developed specific expertise in the field of GPCRs, CAPABILITIES AND PLATFORM ion channels and other cell Evotec has established its antibody surface proteins. Ligand binding, screening platform EVOmAb to target translocation, mito­chondrial incorporate functional screening potential, changes in nuclear early in the antibody discovery and morpholo­ gy, cell differentiation selection process. Attractive targets and replication have been probed such as trans-membrane proteins to name just a few of assays which need to be altered in their activity have been performed. Available in order to achieve a meaningful equipment for antibody screenpharma­ cological effect. Common ing ranges from FLIPR384, Tecan antibody discovery campaigns Safire II, Perkin Elmer EnVision, employ affinity-based processes Opera® and others to highly sophisto enrich manageable number ticated systems such as the Q-patch of antibody clones for functional HTX® or PatchLiner©.

47

ANTIBODY GENERATION AND SELECTION Various platforms available either in house (mice), through collaborations or from customers

GENERATE ANTIBODY POOLS against GPCR’s, Ion channels etc. by immunisation, phage display, yeast display, …

Typical output are thousands of clones (B-cells, Hybridomas, Yeast cells) expressing no or low amounts of soluble antibody

AbEXPRESS High parallel expression of antibodies

CELL LINES Ion Channels GPCRs ANTIBODIES 1,000 to >10,000 clones

Integrins Receptors Cytokines

CELL BASED SCREENING PLATFORMS Established at Evotec in combination with target know-how and tools

CELL BASED ASSAYS Ion Channels – Automated electrophysiology GPCR – Ca2+ flux – cAMP/Arrestin HCS – Signaling cascades – Phenotypic changes – Translocation – Differentiation

EQUIPMENT

FLIPR384 FLIPR Tetra Tecan Safire II Perkin Elmer EnVision TopCount Opera™ SRU BIND IonWorks ® Quattro™ Q-patch HTX® PatchLiner ©

SECTION 13

INTEGRATED SERVICES

Craig Johnstone Senior Vice President Drug Discovery and Innovation Efficiency Discovery Chemistry

TARGET ID & VALIDATION Molecular biology and cloning Bioinformatics In vitro target validation In vivo target validation Target deconvolution

In the previous sections of this document, we have laid out a selection of the key technical capabilities and technologies which we have on offer at Evotec, and these capabilities can be accessed as stand-alone services on demand. In addition, through our extensive drug discovery know-how and experience and expert project management, we bring seamlessly integrated drug discovery capabilities to bear on our collaborations.

We understand that each partner has different needs, internal capabilities and capacities, so through in-depth understanding of the project goals and your specific needs, we develop a coherent plan which enables you to leverage the benefits of our large, flexible, high-quality organisation as a one-stop, cost-effective solution, no

SCREENING Assay development and screening (u)HTS High content screening Biophysical methods Electrophysiology In silico screening technologies Fragment-based drug discovery Compound management

matter where the project lies on the gene-to candidate continuum.

Each collaboration has its own individual challenges so at Evotec “we start with the end in mind”, meaning that we consider the intended indication, frequency and route of administration, safety and efficacy demands, early development strategy etc as part of the product profile which in turn impacts directly on the project plan. Our scientists work with our partner’s scientists to select and execute the most promising strategy and technologies to deliver on the project goals. We also build into our processes key decision points that will guide us through the project and continually measure our progress against these. Using this

HIT-TO-LEAD

LEAD OPTIMISATION

Disease biology and target class expertise Medicinal, synthetic & computational chemistry Library design, procurement and synthesis High-throughput chemistry Protein-ligand crystallography and structure-based drug design In vitro & in vivo pharmacology In vitro ADME, rodent PK & bioanalysis Cellular selectivity analysis Cellular MoA analysis In silico ADMET

DISEASE BIOLOGY EXPERTISE PROVEN PROGRAMME MANAGEMENT

49

approach, we provide innovative and efficient solutions that are always focused on the needs of our partners whilst minimizing waste. Furthermore, by using Evotec as a single provider of integrated services, you can spend more time thinking about the science and less time managing the interfaces between multiple service providers.

Every single day, Evotec scientists are striving to solve drug discovery problems for their clients. Our scientists have made significant contributions to cutting-edge science throughout their careers and are drawn from a variety of backgrounds.

They have been successful in all major therapeutic areas and target classes. Their ideas, inspiration, creativity, innovation and insightful analyses are all key elements of Evotec’s value proposition, and are critical contributions to successful and productive integrated collaborations. In addition, the conduct of such science is heavily dependent on being able to carry out the practical work smoothly, quickly, and without delays, problems, and frustration.

Our client’s satisfaction, and our performance, is dependent on the effective unification of slick, productive, practical execution with good ideas, problem solving and creativity: we call it “Innovation Efficiency”.

As a result of this unique combination of depth, breadth, knowledge and experience of drug discovery with operational excellence, Evotec has established a successful track record in assisting academic institutions, not-for-profit foundations, biotech and pharmaceutical companies in developing novel therapeutics.

Powerful combination of experience and analysis to extract learning and drive next cycle

Excellent understanding of molecular interactions, structural insight and product properties to enable simultaneous multi-parameter optimisation

ANALYSIS

DESIGN

CYCLE TIME

TEST Rapid, relevant, highcapacity efficacy and non-efficacy testing

MAKE MAKE

Rapid and efficient synthetic execution.

Bespoke, complex molecules and designed libraries / sparse arrays

WHY EVOTEC? Industry-leading capabilities in integrated drug discovery World-class suite of compound screening and profiling technologies Proven ability to progress targets to pre-clinical development within 3 years Consistent delivery and innovation for our partners Internal processes optimised to focus on quality and speed allowing for reduced cycle times and thereby accelerating our partner’s discovery efforts Single contracting partner reduces complexity Experienced and proven senior scientists act as project leaders and provide a single point of contact to our partner organisations Expert advice on overall project strategy Securing IP protection and supporting grant applications

SECTION 14

TRACK RECORDS AND CLIENTS

Evotec, with its broad expertise and services, its repeat business and strategic alliances, is perfectly positioned to be your drug discovery services provider of choice. We are aware of the fact that it takes much more than just the highest quality services to develop new drugs, but nevertheless this is the most im­ portant starting point. Evotec delivers an industrialised, high-tech, and comprehensive infrastructure to our partners.

Evotec is one of the few drug discovery businesses that can execute a comprehensive outsourcing strategy.

Evotec provides its customers with access to the Company’s best-inclass fully integrated drug discovery process. In significant largescale, multi-target deals, Evotec’s customers work with our scientists to progress discovery programmes from target through to lead optimisation using the Company’s integrated drug discovery platform.

Evotec has a broad client base ranging from venture capital firms to virtual biotechs to large pharmaceutical companies. We are a truly global company with clients in the United States, Europe and Japan. We have a loyal client base with many long-term agreements as well as regular repeat clients that keep coming back with project after project as Evotec delivers the drug discovery solutions and value they need to drive their discovery programs forward.

Integrated chemistry and biology team at Evotec worked in collaboration with BI scientists, various targets and therapeutic focus, research funding, milestones and royalties

Integrated chemistry, biology and DMPK team at Evotec, working with UCB scientists across various sites on multiple targets, minimum 3 years, research funding, milestones and royalties

Integrated five-year multi-target collaboration in the field of endometriosis, deal structure includes upfront payment plus milestones and royalties

Integrated chemistry and biology team at Evotec, including high-content assays and lead optimisation, recently extended to 2016 (started in 2006)

»Best-in-class alliance work – fruitful for both parties«

»Evotec is the ideal partner to provide the resource bandwidth and drug-hunting expertise«

»Our new collaboration with Evotec will perfectly complement our activities in this field of high unmet medical need«

»We are impressed

by Evotec’s breadth of drug discovery expertise«

51

Imprint Editor: Evotec AG; Chief Editor: Michael Bayer Content: Mario Polywka, Steven Hutchins Design: alessandridesign, Thomas Piribauer Programming: bgcc

Contact us:

Mario Polywka Chief Operating Officer mario.polywka@evotec.com

Steven Hutchins Executive Vice President, Business Development steven.hutchins@evotec.com

Pharmaceutical Accounts

North America

Christophe Muller

Ryan Brady

Vice President, Business Development Key accounts christophe.muller@evotec.com

Vice President, Business Development Venture Capital, Foundations & Not for Profits ryan.brady@evotec.com

Europe

Jeff Naroian Hermann-Josef Kaiser

Business Development Manager Cell and protein production hermann-josef.kaiser@evotec.com

Vice President, Business Development East Coast jeff.naroian@evotec.com

John Kamins Richard Law

Vice President, Business Development, Europe richard.law@evotec.com

Vice President Business Development Proteomics and biomarkers john.kamins@evotec.com

Ashley Rae Kark

Business Development Manager Cell and protein production ashleyrae.kark@evotec.com

Japan Masahiko Otani

Vice President Business Development, Japan masahiko.otani@evotec.com