Eurofins Pharma Discovery Services

Neuropathic pain models:

Chemotherapy-induced, (Rat)

• Reduction of Paclitaxel-induced allodynia.
  
• Allodynia is induced by intraperitoneal injections of Paclitaxel (Taxol, 2 mg/kg) on 4 alternating days (Days 1, 3, 5, and 7).
  
• Rats are preselected (clear presence of allodynia) for experimentation only if the nociceptive response 28 days after Paclitaxel injection (pre-treatment) is reduced by 10 grams of force relative to the response of the individual paw before Paclitaxel challenge. Test substance and vehicle are administered orally to groups of 10 animals one hour before the level of allodynia is again determined (post-treatment).
  
• Acute Assay: In-Life completion in 2-4 weeks from sample receipt
  
• For Subacute Assays: 6 weeks to 3 months
  

Spinal Nerve Ligation, (Chung Model, Mouse & Rat)

• Compound: Gabapentin
  
• Allodynia is induced by L5 and L6 spinal nerve ligation.
  
• Rats are preselected (clear presence of allodynia) for experimentation only if the nociceptive response 14 days after nerve ligation (pre-treatment) is reduced by 10 grams of force relative to the response of the individual paw before nerve ligation (pre-ligation). Test substance and vehicle are administered orally to groups of 10 animals one hour before the level of allodynia is again determined (post-treatment).
  
  • Acute Assay: In-Life completion in 2-4 weeks from sample receipt
    
  • For Subacute Assays: 6 weeks to 3 months
    

Chronic Constriction Injury / Sciatic Nerve Ligation, (Bennett Model, Rat)

• Allodynia is induced by sciatic nerve ligation
  
• Animals are preselected (clear presence of allodynia) for experimentation only if the nociceptive response 7 days after nerve ligation (pre-treatment) is reduced by 10 grams of force relative to the response of the individual paw before nerve ligation (pre-ligation). Test substance and vehicle are administered orally to groups of 8 animals one hour before the level of allodynia is again determined (post-treatment).
  
• Acute Assay: In-Life completion in 2-4 weeks from sample receipt
  
• For Subacute Assays: 6 weeks to 3 months
  

STZ-induced Diabetes, (Rat)

MIA-induced osteoarthritis pain

Sciatic Nerve Crush Injury

Epigenetics, an increasingly popular field of drug discovery, is the study of heritable changes in gene expression that occur without changes in the DNA sequence. The epigenome is considered to be in constant flux and is responsive to environmental and pharmaceutically active molecules. Epigenetics-related enzymes and proteins catalyze the addition or removal of an array of covalent modifications on histone and non-histone proteins. For these reasons, epigenetics is becoming an increasingly important realm for drug therapies. Indeed, several drugs directed towards epigenetic modifying enzymes are already on the market with more in the clinic to treat cancer.

In 2012, ahead of all other CROs, Eurofins recognized the industry's upcoming need for a comprehensive epigenetics program containing all major families. To meet this growing need, we began to develop a range of target based assays using biologically relevant substrates (nucleosomes, histones, and peptides) to create the most meaningful assays. The majority of our proteins/substrates are produced in-house to ensure the highest quality of proteins resulting in high batch to batch reproducibility.

Eurofins' epigenetic toolbox is based on several technology platforms and allows for your choice of technology and/or substrates, providing added flexibility to meet all of your drug discovery needs.

Advantages of Eurofins' Epigenetics Program:

• Largest target and assay list in the industry, with over 160 epigenetic assays to choose from:
  
  • >50 Writers, including: SMMT, Kinases, DNMT, HMT and HAT
    
  • >40 Erasers, including: Demethylases, HDAC / sirtuin and DUB
    
  • >45 Readers, including: Bromodomain and Methyl readers
    
  • >17 Off-target and cell-based assays, including acetylation and methylation studies
    
• Compatible with screening in medium- or high-throughput
  
• All assays set up according to NIH guidelines
  
  • Employing biologically-relevant co-factors and substrates
    
  • Assay optimization includes apparent Km identification for assay constituents and assay linearization
    
• Assays can be modified to meet client needs
  
• Several technology platforms cover the epigenetic portfolio:
  
  • Gold standard filter-based radioactivity
    
  • RapidFire Mass Spectrometry
    
  • AlphaScreen
    
  • TR-FRET
    
• Customer tailored assays to supplement our off-the-shelf assay portfolio
  
  • A specific protein can be cloned, expressed, purified and have a corresponding enzymatic or binding assay developed using a selected technology platform
    

Transporters are membrane proteins that control the influx and efflux of essential nutrients, ions, neurotransmitters, cellular waste, environmental toxins, and other xenobiotics such as drugs. Drug transporters are composed of two major super families: ABC (ATP binding cassette) and SLC (solute carrier) transporters.

ABC transporters mediate unidirectional efflux and are primarily active transporters as they directly rely on ATP hydrolysis to pump their substrates across membranes. They play major roles in hepatobiliary and urinary excretion of drugs from the blood to the lumen, intestinal absorption of drugs, and in brain blood barrier penetration of drugs.

SLC transporters mediate either drug uptake or efflux and can be facilitated transporters or secondary active transporters, which transport drugs using energy produced by transmembrane movement of other solutes in accordance with their concentration gradient. They play major roles in hepatic and renal uptake, and urinary excretion. Some SLC transporters, such as those for neurotransmitters, are also important drug targets themselves with clinical drugs treating various psychiatric disorders.

Transporters play a significant role in a drug's clinical efficacy and safety. Eurofins, through the expertise of Cerep and Panlabs, has developed robust transporter uptake and inhibition assays to allow quick and cost-effective screening of new molecular entities (NMEs) for their interactions with these transporters.

Advantages of transporter screening with Eurofins:

• Cell based assays to provide functional characterization of compound activity
  
• Key drug transporters and neurotransmitter transporters are covered
  
• Drug transporter assays meet regulatory requirements
  

The exact mechanisms of a disease can often be poorly understood. When there is little understanding of the disease pathology, in vitro phenotypic assays may provide more insight than target-based screening alone. Phenotypic screening allows researchers to identify new chemical entities and biomarkers without prior knowledge, or bias, of the target(s) underlying the response. Equally important, phenotypic assays provide a systems-based approach to assess compound liability.

Our high content imaging expertise enables the detection of robust phenotypic responses and is a powerful orthogonal approach to target-based screening. With a diverse portfolio consisting of our OncoPanel™, ImmunoSignal™ and functional toxicity service platforms, Eurofins provides you with the phenotypic results you're looking for.

Understanding the absorption, distribution, metabolism, excretion (ADME) and toxicity (Tox) properties of your compounds is critical in the drug discovery process and can be evaluated by a series of in vitro assays and in vivo studies.

There is a significant advantage to obtaining in vitro ADME data as early as possible in the drug discovery process. For example, hepatic metabolism is a primary determinant of pharmacokinetic behavior and rapid first-pass metabolism is a major cause of low bioavailability. In vitro ADME results serve as good indicators as to whether a compound will be bioavailable, thus these results can assist to eliminate problematic compounds early in discovery and be a useful guide for later stage studies.

Drug toxicity is often the major reason for the withdrawal of an approved drug from the market, with hepatic toxicity and cardiac toxicity being the top two most frequent reasons. While formal toxicity testing is traditionally completed during a late preclinical phase, it has become obvious that a failure at this stage results in significant setbacks and economic loss.

Our assays are designed with the intent to meet regulatory requirements and are suitable for evaluating ADME-Tox properties early in the drug discovery process. With Eurofins cost-effective ADME-Tox assays, we help our clients to eliminate failures earlier in the drug discovery process while liberating them from having to invest internal time and capital to conduct these studies themselves.

ADME-Tox assays available from Eurofins:

• In vivo PK
  
• In vitro bioavailability/ADME
  
  • Solubility / Protein binding / Blood partitioning
    
  • Absorption
    
    • Permeability
      
    • P-gp substrate
      
  • Metabolism
    
    • Blood and plasma stability
      
    • Intrinsic clearance
      
    • CYP- and UGT-phenotyping
      
• Drug-drug interactions
  
  • CYP inhibition and induction
    
  • UGT inhibition
    
  • transporter inhibition
    
• Metabolite identification / Bioanalytical support
  
  • Metabolite identification
    
  • UGT and glutathione conjugates
    
  • Quantitative bioanalysis
    
• In vitro Toxicity
  
  • Genotoxicity
    
    • In vitro micronucleus
      
    • Ames
      
  • Cardiotoxicity
    
    • hERG Channel Screening
      
    • CardiacProfiler™
      
    • Cardiotoxicity by human stem cell derived cardiomyoctes
      
  • Hepatotoxicity
    
  • Nephrotoxicity
    
  • Organtoxicity
    

Eurofins has the experience and skills needed to bring you early stage pharmacokinetic (PK), oral bioavailability and blood-brain barrier (BBB) data to evaluate your new molecular entity. PK and BBB studies are prerequisite for interpreting preclinical efficacy and toxicology results. Using qualitative measurements of drug exposure, we can provide a solid interpretation of preclinical efficacy. PK data can also help in species selection and design of preclinical toxicology studies.

Eurofins will work with you to understand the physicochemical properties of your test material to develop a dosing and sample collection strategy that will provide you with the most meaningful data. We can obtain blood and/or tissue (e.g. brain) samples following dose administration (IV, PO, IP, IM, SC, IN, and ICV, also short- or long-term continuous infusions) and both serial sampling for PK and parallel sampling for PK, BBB or combined PK-BBB are available. PK/PD studies can be combined using our broad portfolio of pharmacodynamics models.

PK/BBB dosing and sampling studies can be paired with standard or custom bioanalytical methods to provide a concentration versus time curves to provide a complete pharmacokinetic profile for your compound.

Advantages of in vivo pharmacokinetic and blood/brain barrier studies with Eurofins:

Extensive experience with multiple routes of administration, including long-term continuous infusion
Proficient in performing PK and BBB studies
Combined PK/BBB sampling and bioanalytical studies
Renal and biliary clearance
Experience you can trust: >25 years running in vivo preclinical services with a staff averaging 15 years of experience
Long range of efficacy and toxicity studies can be conducted at the same provider

Eurofins has built its reputation as a comprehensive, all-in-one provider of discovery assays to take your project from library to preclinical candidate. While we have worked diligently to ensure our assay catalog has everything necessary to support your drug discovery program, we also understand that standard off-the-shelf assays may not have the parameters you require.

Fortunately, Eurofins is able to provide custom assays by modifying existing assays and creating new assays that are tailored to your needs. Our dedicated scientists, who have developed thousands of binding, enzymatic, cell based and in vivo assays, are always available to help work through your assay design and provide a step-by-step plan for your experiment. By trusting Eurofins with your custom assay needs, you're ensuring your project has the greatest chance for success.

Some of the available platforms to design your custom assays around:

High Content Imaging
MD ImageXpress® Micro
Electrophysiology platforms
IonFlux™ HT
Mass spectrometers
Agilent RapidFire 365
Plate readers
FLIPRTETRA® plus ICCD system
Perkin Elmer EnVision®
BMG PHERAStar® FS
Multiplex immunoassays
MSD® Sector® Imager
Scintillation counting
PerkinElmer 1450 MicroBeta® TriLux
Automation and liquid handling
Labcyte Echo®
Biomek® FXP / NX
Agilent BioCel 1200
Cell preparation
MaxCyte® STX®
Vi-CELL®

Proteases are a class of enzymes which perform proteolysis, the mechanism of digesting or cleaving long chain proteins into smaller fragments by severing the peptide bonds that hold them together. Trypsin, a serine protease secreted by the pancreas, is commonly used in cell tissue culture to cleave the bonds that cells use to adhere to a flask.

Proteases also play a major role in disease. HIV-1 protease is essential to the viral replication process. MMP-9, a matrix metallopeptidase, plays a role in angiogenesis and is a therapeutic target for cancer. Because of their significance in the pathology of disease, proteases are a relevant drug target class.

Our broad portfolio of over 50 proteases spanning across the serine, cysteine, aspartic, and metalloprotease families was designed with you in mind to facilitate your drug discovery process and provide you with the answers you need.

Over 50 targets to support your research, including these families and targets:

• Families
  
  • Metalloprotease
    
  • Caspase
    
  • Cathepsin
    
• Targets of interest
  
  • HIV-1 protease
    
  • Proteasome
    
  • DDP4...and many other peptidases
    

Gastrointestinal (GI) disease is characterized by abnormalities in the gastrointestinal tract which includes the esophagus, liver, gallbladder, stomach, pancreas, intestines and colon. Inflammation in these areas can cause a variety of GI diseases or dysfunctions and can be difficult to diagnose. According to the CDC, in the United States alone, inflammatory bowel disease (Crohn's disease or ulcerative colitis) affects over 1 million people. Eurofins, through the expertise of Panlabs, has been conducting in vivo testing for both side-effect profiling and efficacy in GI disease for over 25 years.

Our models allow examination of GI motility and colonic function or to test agents designed to prevent a variety of GI related diseases. We also employ a broad range of outcome measures to assess GI safety and/or therapeutic efficacy of novel test articles. By providing the best models, Eurofins looks to be your partner of choice in helping you to discover the next exciting treatment for gastrointestinal disease.

Advantages of preclinical GI studies with Eurofins:

• Major therapeutically relevant models, including:
  
• Gastric acid secretion and ulcers
  
• Mucosal irritation
  
• Inflammatory bowel disease (IBD)
  
• Irritable bowel syndrome (IBS)
  
• Emesis, including 5-HT emetic models
  
• Companion services can monitor blood or organ exposure and biomarker analysis for PK/PD assessment

Eurofins Metabolic and Endocrine Models

With today's high caloric diets and sedimentary life style, increasing attention has been focused on remedying associated metabolic disorders, naming obesity and diabetes. In early stages, these issues manifest in metabolic syndrome, a complex disease which is characterized by obesity, insulin resistance, pre-diabetic fasting glucose levels, fatty liver, and low HDL cholesterol levels. Left untreated, metabolic disorders can greatly increase the risk of stroke, type II diabetes, and cardiovascular disease.

Eurofins offers a complete list of qualified models for metabolic disorder, including Type I (induction by streptotozocin) and Type II diabetes models. The type I models allow examination of agents acting on insulin-independent pathways and affording end organ protection from diabetes, while the Type II models are conventional wildtype or genetic variants (db/db, ob/ob or ZDF) used with or without modified test diets to explore the impact of pharmacological agents on glucose metabolism and/or obesity. Diabetic complications in humans include organ-specific microvascular damage with sequelae such as nephropathy, and we offer a panel of chronic kidney disease models, as well as models of acute and chronic hepatic injury.

We are able to expertly provide outcome measure of glucose tolerance or insulin tolerance test, as well as plasma lipid profile and measurements of serum insulin, glucagon, leptin, adiponectin, free fatty acid, cytokines, and other relevant biomarkers. Our in vivo testing expertise can help bring your drug from the discovery phase all the way to clinical trials.

Advantages of preclinical metabolic and endocrine studies with Eurofins:

>40 therapeutically relevant models, including:
* Food intake
* Cholesterol
* Glucose
* Estrogen / androgen function
* Renal function
* Renal and hepatic injury
* Models are qualified in a dose-dependent manner with approved benchmark positive controls
* Experienced running combination studies
* Companion services can monitor blood or organ exposure and biomarker analysis for PK/PD assessment

Eurofins Metabolic and Endocrine Models

With today's high caloric diets and sedimentary life style, increasing attention has been focused on remedying associated metabolic disorders, naming obesity and diabetes. In early stages, these issues manifest in metabolic syndrome, a complex disease which is characterized by obesity, insulin resistance, pre-diabetic fasting glucose levels, fatty liver, and low HDL cholesterol levels. Left untreated, metabolic disorders can greatly increase the risk of stroke, type II diabetes, and cardiovascular disease.

Eurofins offers a complete list of qualified models for metabolic disorder, including Type I (induction by streptotozocin) and Type II diabetes models. The type I models allow examination of agents acting on insulin-independent pathways and affording end organ protection from diabetes, while the Type II models are conventional wildtype or genetic variants (db/db, ob/ob or ZDF) used with or without modified test diets to explore the impact of pharmacological agents on glucose metabolism and/or obesity. Diabetic complications in humans include organ-specific microvascular damage with sequelae such as nephropathy, and we offer a panel of chronic kidney disease models, as well as models of acute and chronic hepatic injury.

We are able to expertly provide outcome measure of glucose tolerance or insulin tolerance test, as well as plasma lipid profile and measurements of serum insulin, glucagon, leptin, adiponectin, free fatty acid, cytokines, and other relevant biomarkers. Our in vivo testing expertise can help bring your drug from the discovery phase all the way to clinical trials.

Advantages of preclinical metabolic and endocrine studies with Eurofins:

>40 therapeutically relevant models, including:
* Food intake
* Cholesterol
* Glucose
* Estrogen / androgen function
* Renal function
* Renal and hepatic injury
* Models are qualified in a dose-dependent manner with approved benchmark positive controls
* Experienced running combination studies
* Companion services can monitor blood or organ exposure and biomarker analysis for PK/PD assessment

Custom assays to support your drug discovery project

Eurofins has built its reputation as a comprehensive, all-in-one provider of discovery assays to take your project from library to preclinical candidate. While we have worked diligently to ensure our assay catalog has everything necessary to support your drug discovery program, we also understand that standard off-the-shelf assays may not have the parameters you require.

Fortunately, Eurofins is able to provide custom assays by modifying existing assays and creating new assays that are tailored to your needs. Our dedicated scientists, who have developed thousands of binding, enzymatic, cell based and in vivo assays, are always available to help work through your assay design and provide a step-by-step plan for your experiment. By trusting Eurofins with your custom assay needs, you're ensuring your project has the greatest chance for success.

Some of the available platforms to design your custom assays around:

• High Content Imaging
  
  • MD ImageXpress® Micro
    
• Electrophysiology platforms
  
  • IonFlux™ HT
    
• Mass spectrometers
  
  • Agilent RapidFire 365
    
• Plate readers
  
  • FLIPRTETRA® plus ICCD system
    
  • Perkin Elmer EnVision®
    
  • BMG PHERAStar® FS
    
• Multiplex immunoassays
  
  • MSD® Sector® Imager
    
• Scintillation counting
  
  • PerkinElmer 1450 MicroBeta® TriLux
    
• Automation and liquid handling
  
  • Labcyte Echo®
    
  • Biomek® FXP / NX
    
  • Agilent BioCel 1200
    
• Cell preparation
  
  • MaxCyte® STX®
    
  • Vi-CELL®

Develop difficult targets and orthologs to meet your drug discovery needs

Outsourcing your stable cell line development to Eurofins will enable you to free up necessary resources while guaranteeing the success your drug development program needs. We have successfully developed hundreds of human GPCR and ion channel stable cell lines and related orthologs, bringing time-tested experience to the table. Our stable cell line development FlexLab option allows you to choose from CHO, HEK, or our proprietary Chem-1 host, a rat hematopoietic, adherent cell line or provide your own.

Our experienced scientists use our proprietary expression vectors to generate cells producing the most functionally active recombinant proteins. Our milestone driven projects give you the flexibility to take projects from proof of concept transient transfections, to selected polyclonal cells and all the way to stability tested monoclonal cell lines.

Advantages of Eurofins' Custom Cell Line Development:

• Extensive experience with both GPCRs and Ion Channels
  
• Choose from transiently transfected cells or stable cell lines
  
• Access to MaxCyte STX electroporation system for robust multi-plasmid transfection for bulk transiently-transfected frozen cells
  
• Ability to qualify cells for assay performance on a large range of platforms

If your intent is to use protein for enzymatic assays, as substrates in biophysical assays (e.g., surface plasmon resonance (SPR)), for crystallography or nuclear magnetic resonance (NMR), we can produce the proteins with the specifications that match your application. Our protein expression experts share a wealth of experience in recombinant protein expression and have produced >400 protein products, all of which are made to demanding control parameters (typically including: ≥ 80% protein purity, ≥ 0.5 mg/L yield, and specified activity for protein kinases).

We have a robust manufacturing process that is highly controlled and documented at every stage with a manufacturing batch record. As a milestone, we can produce a batch of pilot scale material for your in-house testing before you commit to a large scale project.

Our custom protein production and purification skills are unmatched and can be applied to suit your unique needs. We offer bulk manufacturing as well as wild type, mutant, and ortholog proteins. Contact us today to speak with our protein specialists.

Protein expression and purification capabilities:

• Expression systems
  
  • E. coli
    
  • Baculovirus
    
  • Mammalian (CHO, HEK)
    
• Purification expertise
  
  • IMAC
    
  • Ion exchange
    
  • Gel filtration
    
  • Affinity chromatography
    
• Expression scales and capacities
  
  • Shake flask (50mL - 1L) - 700L/month capacity
    
  • Benchtop bioreactors (5L and 10L) - 250L/month capacity
    
• Expression scales and capacities
  
  • SDS Page
    
  • Purity estimation
    
  • Mass spectrometry
    
  • Western blot
    
  • Activity assay (if available)

Approval and registration of drugs requires a comprehensive assessment of their genotoxic potential. Genotoxicity testing is an integral component of regulatory toxicity evaluation in most countries. Since no single test is capable of detecting all relevant genotoxic end-points, a battery of in vitro and in vivo tests for genotoxicity is recommended by regulatory agencies. The recommended standard test battery includes in vitro tests for gene mutation in bacteria (Ames test) and in mammalian cells (mouse lymphoma assay) or an in vitro test for chromosomal damage in mammalian cells (in vitro micronucleus test or in vitro metaphase chromosomal aberration assay). Additionally, an in vivo test for chromosomal damage (in vivo micronucleus or in vivo chromosomal aberration assay) is required by regulatory agencies as part of an IND application.

Recently, assessment of genotoxicity testing has evolved towards earlier stages of drug discovery in order to identify genotoxic liabilities at a time when changing lead series will have a relatively low impact on a project's timeline and cost. Assays such as the Ames fluctuation test and the in vitro micronucleus assay (for the assessment of the in vitro chromosomal aberration) offered by Eurofins are routinely used for screening compounds.

Advantages of Eurofins' Genotoxicity Assays:

• Ames fluctuation test and in vitro micronucleus assays are offered to meet ICH S2(R1) recommendations
  
• Assays require small amount of compound and provide rapid turnaround time (10 business days)
  
• The Ames fluctuation test utilizes 4 Salmonella typhimurium tester strains (TA98, TA100, TA1535 and TA1537) to detect both frameshifts and base-pair substitutions
  
  • Testing can be performed with and without metabolic activation by rat liver S9 fraction
    
• Bacterial cytotoxicity assays are conducted concurrently with Ames test to confirm the negative Ames results
  
• The in vitro micronucleus assay uses automated scoring by High Content Analysis (HCA) which provides objective and consistent data when compared with manual scoring.

Toxicity testing is mandated by regulatory agencies for IND submission and is typically performed during the preclinical phase of discovery. However, uncovering toxicity at such a late stage means that considerable investment and time has been lost, jeopardizing the project. Eurofins has developed a reliable high-throughput model of multiplexed toxicity testing which provides a quick and cost-effective option suitable for early liability assessment, when changing lead series has minimum impact on the project. By utilizing imaging expertise with five human primary cell lines, we can provide a sensitive in vitro cell-based method to aid in lead selection and optimization by identifying potential tissue-relevant toxicity earlier in the drug development process.

Our organ toxicity services employ primary cells derived from human donor tissue that are purified and cryopreserved. Primary cells such as the Normal Human Dermal Fibroblasts (NHDF), Human Renal Proximal Tubular Epithelial Cells (HRPTEpiC), Human Umbilical Vein Endothelial Cells (HUVEC) and Human Umbilical Mesenchymal Stem Cells (HUMSC) included in this panel are able to propagate in for a short number of passages before they stop dividing. These cells are banked at a low passage number and proliferate in culture pre- and post-plating. The Human Primary Hepatocytes (HPH) are plated directly from thawed cryovials and do not proliferate in culture.

Our assays employ high-resolution cell imaging to provide more sensitive detection of cell viability determined by proliferation inhibition or cell loss. Our services can be run with a single cell type, the pre-validated primary cell panel, or cell lines chosen through collaboration with our expert scientists who have experience running cytotoxicity assays with hundreds of human cell lines.

Advantages of Eurofins' Organ-Toxicity in Human Primary Cell Panel:

• Pre-validated human cells harvested and purified from single donor source are used for reproducible assays
  
  • Normal Human Dermal Fibroblasts (NHDF)
    
  • Human Renal Proximal Tubular Epithelial Cells (HRPTEpiC)
    
  • Human Umbilical Vein Endothelial Cells (HUVEC)
    
  • Human Umbilical Mesenchymal Stem Cells (HUMSC)
    
  • Human Primary Hepatocytes (HPH)
    
• Flexible project scope from single cell type to full cell panel
  
• Low compound requirement - as little as 300 μl of a 20 mM solution is needed for testing at a final assay concentration of 100 μM
  
• Relevant reference compound and vehicle controls are included on each plate

Establish the metabolic profile of a promising compound early in the drug-discovery process

Metabolic profiling of therapeutic compounds at early stages in the drug discovery process is of increasing importance. Drugs that undergo metabolism in vivo may produce pharmacologically active or chemically reactive metabolites which can produce unexpected effects or potential toxicities. The recent FDA Guidance for Industry on Safety Testing of Drug Metabolites1 highlights the relevance of in vitro metabolite profiling early in drug development, as metabolites which are unique to or disproportionate in humans may require additional toxicological studies.

In addition to an extensive panel of in vitro metabolism screening assays, Eurofins is able to perform studies to thoroughly characterize the metabolic products of a drug compound in vivo or in vitro and compare metabolic profiles across species. The key element in metabolite identification studies is the use of state-of the art mass spectrometry instrumentation for sensitive and accurate detection of metabolic products and software tools to efficiently process the complex datasets that are produced from these analyses.

Eurofins employs a Waters Xevo G2 QTOF MS system combined with Acquity UPLC separation to provide high mass accuracy and excellent sensitivity in full scan MS and MS/MS modes. The acquisition method on the Xevo collects precursor and product ion data in two distinct but parallel acquisition functions via rapid switching, so that complete metabolite identification data can be obtained from a single LC injection. If necessary, follow-up selected reaction monitoring or product ion or neutral loss scans can be performed on a triple-quadrupole MS system in order to obtain corroborating data for putative metabolites.

By partnering with Eurofins, you'll be able to better understand your drug candidate's metabolism and stability and obtain a more comprehensive assessment of that compound's metabolic profile.

Flexible options to meet your research needs
Metabolism data on test compounds can be obtained through a choice of experiments, depending on the level of data depth and interpretation required:

• Enhanced Metabolic Stability (Metabolic soft-spot analysis)
  
  • Liver microsomes, S9, hepatocytes
    
  • Phase I and/or Phase II metabolites
    
    • % remaining and (if multiple timepoint) half-life of parent compound
      
    • Extracted ion chromatograms of parent and major metabolites
      
    • Proposed metabolic transformation for each observed metabolite
      
    • Tabulated list of metabolite masses, peak areas, and retention times
      
• Metabolite Identification
  
  • Liver microsomes, S9, hepatocytes
    
  • Phase I and/or Phase II metabolites
    
  • All of above plus:
    
    • MS/MS spectra of parent compound and observed metabolites
      
    • Structural interpretation of productions of parent and metabolites
      
    • When possible, molecular formula and proposed structures of metabolites.
      
• In vivo Metabolite Analysis
  
  • Plasma or tissue from in-life studies
    
  • Circulating Phase I and Phase II metabolites
    
  • Either of above options

Safety related issues and drug toxicity is a major reason why promising leads don't advance and candidates fail curing clinical development. The 'fail early' mantra in drug discovery was born out of the reality that late-stage failures result in significant setbacks and economic loss.

For early safety and toxicity assessment, Eurofins is the go-to-provider with over 35 years of in vivo safety and early toxicity testing. Combined with our in vitro pharmacology and cell based in vitro toxicity assays services, our in vivo safety services can provide a comprehensive risk profile for your leads at a stage when changes can still be made without a major impact to your project.

We offer a complete list of qualified safety related in vivo models to detect the most commonly observed adverse events in CNS, cardiovascular, respiratory, renal, metabolic, and gastrointestinal functions using a broad panel of industry-standard outcome measures to build a toxicokinetic (TK) profile. We can also provide specialized models, such as our emesis model, to address specific project needs.

Advantages of early, non-GLP safety screening with Eurofins:

• Maximum Tolerated Dose
  
• 7-day or 28-day tox (non-GLP)
  
• PK + tissue exposure, urinary excretion
  
• Cardiovascular
  
  • Blood pressure and heart rate
    
  • Postural hypotension and tilt response
    
  • Arrhythmia and QTc interval
    
• Neurological and behavioral effects
  
  • Functional Observation Battery (Irwin) profile
    
  • Convulsions
    
  • Autonomic signs
    
  • Locomotor activity and coordination (Roto-rod)
    
• Gastrointestinal and metabolic effects
  
  • GI motility
    
  • Emesis
    
  • Gastric effects
    
  • Body weight and food consumption
    
• Hepatic and renal function
  
• Respiratory monitoring
  
• Ancillary services: clinical chemistry, hematology, blood gases, gross pathology, histology and macroscopic and microscopic necropsy

Respiratory diseases affect the upper respiratory tract and can result in difficulty breathing, decreased lung capacity, asthma, emphysema and a host of other conditions. Left untreated, respiratory diseases can result in chronic breathing issues and become potentially life threatening.

Eurofins recognizes the need for effective and relevant in vivo models for respiratory conditions. In order to help you bring the next potential respiratory treatment to clinical trials, we offer a number of relevant qualified models to help you understand you candidate's effectiveness.

With more than 25 years of experience, our experts can provide you the data to give you confidence to take the next step in your project.

Advantages of preclinical respiratory studies with Eurofins:

• Therapeutically relevant models, including:
  
  • Allergic reaction and antagonism of pro-inflammatory mediators
    
  • Pulmonary inflammation and fibrosis
    
  • Antitussive / cough suppression
    
  • Bacterial pneumonia
    
  • LPS induced neutrophilia
    
• Multi-faceted outcome studies
  
  • Cytokine biomarker detection in plasma or bronchoalveolar lavage fluid (BALF) using ImmunoSignal™ services
    
  • Blood gases and respiration rate, tidal volume
    
  • Minute volume employing whole body plethysmography
    
  • Detailed analysis of respiratory mechanics
    
• Models are qualified in a dose-dependent manner with approved benchmark positive controls
  
• Experienced running combination studies
  
• Companion services can monitor blood or organ exposure and biomarker analysis for PK/PD assessment

Xenograft and syngeneic and in vivo tumor models

With over 25 years in the industry, Eurofins has refined our skills to excel at providing syngeneic and human tumor xenograft models. Our experience is unparalleled in providing in vivo models of efficacy in murine or human hematologic or carcinoma malignancies.

Our oncology services also feature custom model development for clients' target-specific cell lines as orthotopic, disseminated/metastatic or subcutaneous xenografts, using any of the 300 cell lines from our cell based phenotypic assays found in our OncoPanel™ services. Our ability to establish custom xenograft models from patient-derived tissue is a testament to our skills and experience in oncology services.

Trusting Eurofins with your preclinical oncology studies helps you understand your candidates' efficacy, so you can be confident entering clinical development.

Advantages of profiling with in vivo oncology models:

• 20 off-the-shelf models available including:
  
  • Xenograft models
    
  • Syngeneic models
    
• Tissues covered
  
  • Brain, breast, colon, kidney, lung, blood, pancreas, prostate, skin
    
• Experienced running combination studies
  
• Highly customizable capabilities
  
  • Model development available for target-specific cell lines
    
  • Patient-derived tumor xenografts
    
• Companion services can monitor plasma, tissue or tumor exposure and biomarker analysis for PK/PD assessment

The central nervous system (CNS) is prone to many diseases and pain-related disorders, especially prevalent during aging. The importance of CNS disorders is indicated by the severity of many of the diseases associated with it. These diseases include depression, epilepsy, psychosis, multiple sclerosis, and many other common, yet difficult-to-treat diseases. Eurofins, through the expertise of Panlabs, has been conducting in vivo testing for CNS and pain disorders for both efficacy and side-effect profiling for over 25 years.

We have applied our in vivo and neuroscience acumen to develop the most relevant CNS and pain-related models including epilepsy, affective disorders, neuroleptic, and multiple sclerosis, acute, inflammatory and neuropathic pain. We also offer translational models of pain such as osteoarthritis and bone cancer pain, a testament to our superlative skills. Additionally, we offer several pain-associated behavioral measurements such as mechanical allodynia, thermal and mechanical hyperalgesia, as well as cold allodynia.

With our ever-expanding portfolio of in vivo models, Eurofins has the necessary skills and expertise to assist you to evaluate whether your candidates are ready for clinical trials.

Advantages of Eurofins for preclinical CNS and pain studies:

• >50 CNS and pain assay models available
  
• Models include:
  
  • Analgesia / pain
    
    • Acute, inflammatory & neuropathic pain
      
  • Epilepsy / convulsions
    
  • Affective disorders
    
    • Anxiety
      
    • Depression
      
    • Antipsychotic/neuroleptic
      
  • Multiple sclerosis
    
  • Motor activity
    
• Models are qualified in a dose-dependent manner with approved benchmark positive controls.
  
• Companion services can monitor blood, brain, spinal cord and/or CSF exposure and biomarker analysis for PK/PD assessment

The ImmunoSignal™ platform provides a suite of services to understand the immune response to your therapeutic. Our in vitro solutions for discovery pharmacology and preclinical research include multiplexed cytokine analysis and flow cytometry techniques, allowing simultaneous monitoring of pro- and anti-inflammatory markers.

We conduct assays using a unique bank of Peripheral Blood Mononuclear Cells (PBMCs) from donors of diverse ethnical backgrounds, offering genetic diversity. These primary cell banks are from single human donors for a consistent cellular source for cytokine secretion assyes, which is useful for maintaining cross data integrity. Monocytes, T-cells and B-cells have also been isolated from these banks for proliferation and cytokine studies.

Enjoy tailored made solutions for your immunological studies by using our custom services. We can use your specified cell lines or sub-cellular components isolated from white blood cells or design a panel to meet your specific needs.

Whether you decide on an off-the-shelf solution or a highly customized assay, Eurofins has the immunology solution you need to further your drug development program.

Advantage of ImmunoSignal™ services:

• Reliable and predictive with access to a large bank of well-characterized human cells
  
  • Multiple human donors offering genetic diversity
    
  • Characterized for sensitivity and robustness in cytokine or adhesion protein release levels
    
• Expert scientific and technical support for data interpretation
  
• Flexible, custom immune marker panels and testing conditions available
  
  • Access relevant cellular models and biological fluids from animal studies.
    
• Quality, fully validated and quality-controlled
  
• Multiplex analysis and flow cytometry techniques to detect bioananlytes and cellular proliferation and viability
  

Preclinical evaluation of suppression of immunological and inflammatory responses

Through Panlabs, Eurofins has 25 years of experiences creating and working with a diverse set of in vivo efficacy models for inflammatory diseases. These models range from basic models of immune modulation, restoration, suppression and stimulation to specific models of inflammation-induced organ damage. All experimental models are tested in a dose-dependent manner with approved benchmark positive controls.

Our ability to deliver multi-faceted outcome studies, means that you will receive the data that is most appropriate for your program. We routinely measure relevant biomarkers (e.g., cytokines) employing our highly sensitive ImmunoSignal™ services; physiologic outcomes, like airway hyper responsiveness, and swelling; as well as more complex behavioral outcome measures such as scratching or mechanical allodynia or histopathology.

Rely on Eurofins to deliver the most meaningful preclinical data, allowing you to decide whether your candidates' efficacy meets the criteria for full clinical development.

Advantages of preclinical inflammation / allergy studies with Eurofins:

• >30 therapeutically relevant models, including:
  
  • Collagen (CIA) or adjuvant (AIA) induced arthritis
    
  • Colitis
    
  • Pulmonary inflammation
    
  • Allergic reaction
    
  • Allergic/irritant contact or atopic dermatitis
    
  • Septic shock - LPS-induced or cecal ligation and puncture
    
  • Wound healing
    
  • Pancreatitis
    
  • Fibrosis - pulmonary, hepatic or renal
    
• Models are qualified in a dose-dependent manner with approved benchmark positive controls
  
• Experienced running combination studies
  
• Companion services can monitor blood or organ exposure and biomarker analysis for PK/PD assessment

In vitro services to find the next generation drugs to combat infectious disease

With the rising prevalence of drug-resistant microorganisms, effective and potent anti-infective agents are becoming increasingly more relevant in order to treat infectious diseases. Eurofins, through the expertise of Panlabs, identified this need over 25 years ago and has since been working diligently with the pharmaceutical industry to provide a robust portfolio of in vitro and in vivo anti-infective models to support the discovery and development of novel antimicrobial therapies.

Our in vitro portfolio follows the standard industry methods (CLSI) in order to evaluate the antimicrobial activity of your test agents. Our services have unmatched flexibility, allowing for combination testing, determining synergistic, indifferent, or antagonistic effects as well as resistance analysis to determine the rate at which organisms develop a non-susceptible phenotype which can have major clinical implications.

Using a comprehensive collection of over 270 clinically relevant Gram-positive and -negative bacteria (including multi-drug resistant, MDR, organisms), anaerobic bacteria, yeast, and filamentous fungi, Eurofins provides the support and experience that your anti-infective drug program needs to succeed.

Features of in vitro anti-infective services from Eurofins:

• Determination of minimum inhibitory concentration (MIC)
  
  • Minimum Bactericidal Concentration (MBC)
    
  • Antibiotic susceptible organisms
    
  • MDR organisms, clinically relevant resistant mechanisms
    
  • Effect of human and/or mouse serum
    
  • Accelerated, one-week turnaround time is available
    
  • High volume compound testing
    
• Panels for accelerated MIC testing
  
  • Bacterial broad spectrum screen
    
  • Fungal broad spectrum screen
    
  • Neisseria gonorrhoeae - MDR and susceptible strains
    
• Time-kill analysis to identify bactericidal properties
  
• Combination testing to determine synergistic, indifferent or antagonistic effects
  
• Resistance analysis to determine the rate at which organisms develop a non-susceptible phenotype
  
• Membrane lysis - bacteria and red blood cells
  
• Clostridium difficile toxin - cytotoxicity and toxin neutralization
  
• >270 pathogens, including:
  
  • Anaerobic bacteria: C. difficile, P. vulgaris
    
  • Fungi: C. albicans, A. fumigatus
    
  • Gram-negative bacteria: A. baumannii, E. coli, K. pneumoniae, N. gonorrhoeae, P. aeruginosa
    
  • Gram-positive bacteria: E. faecalis, S. aureus, S. epidermidis, S. pneumoniae, S. pyogenes
    
• Companion in vivo services for preclinical anti-microbial assessment

Cancer is a multi-dimensional disease driven by genetic instability and selective pressure to continue to persist. As such, an ever changing tumor presents a formidable challenge to effectively treat. Given this challenge, a strategy that continues to be employed is a multi-prong attack on the tumor to eliminate multiple traits and/or to prevent the tumor from developing resistance mechanism to circumventing treatment.

To support this multi-prong approach to cancer therapy, combinations of therapeutics (small-molecules or biologics) are tested on OncoPanel™ cell lines in a dose response matrix. This data format is ideal for a variety of downstream drug-drug combination analyses including calculation of combination index, isobolographic analysis, and determination of excess over Bliss independence.

Goals of for drug combination studies:

• Increase the efficacy of drugs
  
• Combat drug resistance
  
• Provide long-term tumor control for patients
  
• Increase the therapeutic window of individual chemotherapies
  
• Combine small molecules and biologics

OncoPanel™ services provide multi-parametric drug response data across a panel of 300 genomically diverse cancer cell lines that span 19 different cancer tissues. Therapeutics may be tested in the entire panel or a subset that contains any combination of the 300 available cancer cell lines. Using high content imaging, simultaneous detection of anti-cancer activity and detection of pharmacodynamic biomarkers in a single, multiplexed assay may be provided.

OncoPanel™ services allow a detailed understanding of drug response that cannot be achieved by single endpoint assay profiling. Assay endpoints can be customized to measure up to four biological pathways or processes at once (cell viability, apoptosis, mitotic block and custom markers). Genomic analysis of OncoPanel™ data enables the discovery of predictive biomarkers to help determine which genomic features may predispose patients to therapeutic response.

Cell surface expression of your target antigen can be measured in live cells to correlate antigen expression with therapeutic response in the same assay. OncoPanel™ can also provide quantitative binding analysis of therapeutic antibodies within the same multiplexed assay.

Study design to match your project goals
As assays become more sophisticated, it is often necessary to tailor parameters to the specific question that is being asked. Our experience working with a variety of different molecules, such as kinase inhibitors, epigenetic modulators and large molecule biologics, ensures that we can generate the best study design for your project.

The OncoPanel™ Advantage:

• Univariate genomic analysis to identify predictive biomarkers to correlate sensitivity and resistance
  
  • Genomic data repository: Eurofins, Sanger COSMIC, Broad CCLE
    
  • Univariate genomic analysis includes
    
    • Mutations
      
    • DNA copy number alterations (amplifications, homozygous deletions)
      
    • Differential mRNA expression
      
• >300 genomically characterized cancer lines provides statistical power to uncover and rate predictive biomarkers
  
  • Rigorous cell culture quality control, provides consistent responses and reproducibility
    
• Experienced in testing small molecules, biologics & drug combinations
  
• State-of-the-art High Content Analysis (HCA) instrumentation and automation to deliver objective and more consistent data analysis
  
• Multi-parametric analysis for rich mechanistic data
  
  • Cell viability - decreased cell count using DAPI (nuclear dye) stained cells
    
  • Apoptosis - increased staining with an anti-active caspase antibody
    
  • Mitotic block - increased staining with an anti-phospho-histone-H3 antibody
    
  • Custom markers - project specific endpoints (e.g., PD biomarker)
    
• Flexibility - test compounds in our pre-existing cell line panel or send your own cell lines
  
• Pioneers for image based oncology services, establishing studies in 2007
  

OncoPanel™ 3D – determine activity of compounds on tumor spheroids

OncoPanel™ 3D is a drug profiling platform comprised of ~100 cancer cell line models grown as three-dimensional (3D) tumor spheroids that phenocopy key attributes of human tumors. Drug testing is initiated only after spheroid formation has been confirmed through hypoxia-positive spheroid calling using high-content imaging. This platform provides a means to compare the tumor penetrating properties of a collection of drugs by comparing activity in 2D versus 3D cell culture.

Large molecule biologics

The types of therapeutics used to treat cancer are the most diverse among therapeutic areas. Of growing success and interest has been large molecule therapeutics for oncology, especially humanized monoclonal antibodies (mAbs). MAbs are generally targeted to cell surface receptors, or the ligands of these receptors, to block various oncogenic hallmarks (e.g., uncontrolled cell proliferation). OncoPanel™ services can help profile cell surface antigen expression for a mAb's target and identify drug responsiveness in the same assay

Epigenetic therapeutics

Druggable epigenetic modulators undergo oncogenic mutation and are seen as attractive therapeutic targets. However, inhibition of some epigenetic targets results in a significantly delayed drug response that is not revealed by conventional, short term cell growth assays. To address this issue, we have established two- and three-dimensional assay conditions with extended assay duration to yield robust cell line profiling data. While this assay design was specifically developed with epigenetic targets in mind, other slow-acting therapeutics are likely to reveal themselves employing this extended protocol.

Determine your drug’s similarity to existing drugs with BioPrint®

BioPrint® is a large, homogenous pharmacology and ADME database, which provides a unique resource for supporting the drug discovery decision-making process. The database is composed of three main data sets:

Chemical descriptors (structures and chemical information, 2D- and 3D- descriptors)
In vitro data
Collected and curated in vivo effects of drugs
Our standard BioPrint® profile includes the assays used to explore the properties of >2,500 BioPrint® compounds (consisting of marketed drugs and reference compounds), which establishes individual Pharma-ADME fingerprints for each compound.

BioPrint® positions a new drug candidate in the context of marketed drugs, allowing you to anticipate potential in vivo liabilities, predicting off-target activities, and ADME characteristics (drug profile interpretation). BioPrint® can also help identify secondary therapeutic targets that are not genetically related to a test target (target profile design) for drug repositioning.

Advantages of a BioPrint® profile:

• Includes a panel of:
  
  • 104 binding assays (non-peptide, peptide and nuclear receptors, ion channels and amine transporters)
    
  • 31 enzyme assays (including 10 kinases, 10 proteases and 5 PDEs)
    
• Identify marketed drugs and reference compounds with similar pharmacological profiles or sub-profiles to that of a test compound
  
• Hypothesize clinical effects of new drug candidates based on the well characterized in vivo effects of drugs with similar profiles
  
• Place individual hits on a test compound in the context of similar strength hits on marketed drugs and reference compounds
  

Predict in vivo activity from in vitro data

The Foresight™ similarity analysis service allows you to compare your compound's assay results against the assay results of a large reference set of pharmacologically active compounds. The reference set includes marketed drugs, failed development candidates and reference compounds that have well-characterized activities and have been tested against a series of expert-selected hit and lead screening assays. This service is compound-centric and includes all data generated for your compound.

Using the actual data derived from tests run on your compounds, we determine the five compounds in the reference set with the highest similarity score. The score is computed based on the mathematical correlation of assay test results of your compound to the reference set compound assay results, providing you with an insight into potential in vivo activity.

Advantages of a Foresight™ Profile:

• Choose different mathematical models to test the fit between the test compound and reference set assay results
  
• Determine potential adverse effects and identify possible new indications

Bioavailability of a compound is mainly determined by absorption, distribution, metabolism and excretion (ADME), which can be evaluated by a series of in vitro assays. In small molecule drug discovery the typical goal is to identify a low molecular weight compound which can be effectively administered via an oral route. The fraction of a dose after drug administration that reaches the general circulation intact is defined as its bioavailability.

There is a significant advantage to obtaining in vitro ADME data as early as possible in the drug discovery process. For example, hepatic metabolism is a primary determinant of pharmacokinetic behavior and rapid first-pass metabolism is a major cause of low bioavailability. In vitro ADME results serve as good indicators as to whether a compound will be bioavailable and assists in eliminating failures early in the development process. It also serves as a useful guide for later stage studies.

Eurofins offers a diverse set of in vitro ADME assays which, if applied early in the drug discovery process, can be used to prioritize compounds based on these properties and to select those compounds likely to have high bioavailability for further development.

Assessment assays for bioavailability:

• Aqueous Solubility
  
• Partition Coefficient
  
• Protein Binding / Blood Partitioning
  
• In vitro Absorption
  
• In vitro Metabolism
  
• Caco-2 Permeability
  
• P-gp Substrate Determination

Undesirable ADME properties are major factors causing failures during drug development. To prevent these costly failures from occurring, in vitro screening of potential drug candidates in the early drug discovery phase has been employed as a cost-effective approach to identify compounds with unfavorable ADME characteristics. Eurofins provides a wide variety of metabolism screening assays using assay matrices, including: recombinant cytochrome P450 enzymes (CYPs), recombinant uridine diphosphate-glucuronosyl transferases (UGTs), liver microsomes and hepatocytes from human and animal species.

After entering the body, a drug will be metabolized to one or several more water-soluble metabolites, which can be more readily excreted than the corresponding parent drug. The enzymes that catalyze chemical conversion during the metabolism process are categorized as phase I (mainly CYPs) and phase II (such as glutathione-S-transferases and UGTs) drug-metabolizing enzymes. These enzymes are present at high levels in organs of elimination, such as the liver, intestine, and kidney. Intrinsic clearance assays using liver microsomes or hepatocytes will allow you to estimate the extent of metabolic clearance and first-pass effect for your compounds. Further studies of CYP- and UGT-reaction phenotyping will allow you to understand whether your compound is subject to drug-drug interactions or polymorphic effect.

By partnering with Eurofins, you'll be able to better understand your drug candidate's metabolism and stability and obtain a more comprehensive assessment of that compound's drug profile.

In vitro metabolism screening assays allow identification of compounds that have these favorable characteristics:

• Stability - small first-pass effect to maintain an effective concentration in blood for a reasonable period of time
  
• Metabolized by multiple CYP enzymes and not largely dependent on CYPs that are polymorphically-expressed, such as CYP2C9, CYP2C19, and CYP2D6
  
• Lead to no pharmacologically-active (unless starting as a pro-drug) or chemically-reactive metabolites

Metabolism-mediated drug-drug interactions (DDI) refer to simultaneously administered drugs that interfere with each other's absorption, distribution, metabolism, and elimination (ADME) by changing activities of drug metabolizing enzymes and/or drug transporters.

As an example, drug A may inhibit or induce activity of an enzyme(s) or transporter(s) directly or virtual of it being a substrate of these proteins. As a consequence, another medicine, drug B, that is a substrate of those affected enzymes and transporters will have its PK profiles altered and vise versa.

DDI alters drug's pharmacokinetic (PK) profile and this change in concentration and duration profile may lead to either clinical ineffectiveness or adverse drug reaction in patients that can be severe or fatal. Both the FDA and EMA have issued guidances which recommends that all new drugs be investigated to understand their potential to interact with other drugs.

To address this need, Eurofins provides a wide range of assays to help our clients screen their compounds for drug-drug interactions. Early screening of DDI characteristics of a compound is critical in reducing late-stage failures.

Available DDI Profiling Assays:

• CYP- and UGT-reaction phenotyping
  
• CYP inhibition (reversible and time-dependent) and CYP induction (mRNA level)
  
• UGT inhibition
  
• P-gp substrate
  
• Transporter inhibition

Reversible protein phosphorylation is critical for normal cellular function with perturbations leading to diseases such as cancer. This post-translational modification alters protein function either by relocating proteins, changing protein interaction and/or modifying their enzymatic activity. Phosphatases are the key enzymes involved in removing phosphatase groups and thus reversing the effect of kinase directed phosphorylation. Although lagging behind kinases in maturity as a therapeutic drug class, interest is growing in phosphatases, like PTP-1B, as future therapeutic targets. As with kinases, phosphatase inhibitor specificity is a major challenge for drug discovery scientists.

Through the combined expertise of Cerep, Panlabs, and PhosphataseProfiler™ services, Eurofins has developed a robust, reliable, and well-characterized solution to screen your focused chemical libraries or profile your lead compounds for their inhibitory effects on phosphatases.

Advantages of Phosphatase Profiling with Eurofins:

• Over 40 available assays for selectivity profiling of novel chemical entities
  
• Over 10 years of experience in phosphatase profiling
  
• One week turnaround time available to drive SAR
  
• Single point inhibition data or 10-point IC50 curves
  
• Unmatched robustness and reproducibility

Ubiquitination is a key regulatory mechanism which attaches the small protein modifier ubiquitin to protein substrates, thereby modifying their structure, function, cellular location, or targeting them for destruction via proteolysis. Inappropriate regulation of ubiquitination pathways is linked with a number of human diseases. In recent years, components of these pathways have emerged as a new and relatively untapped class of targets for drug discovery, with applications in cancer, neurodegenerative disorders such as Alzheimer's and Parkinson's disease, viral infection, diabetes, and inflammation.

Eurofins has removed the complexity of drug discovery in this emerging area by providing assays for both ubiquitination and de-ubiquitination, with off-the-shelf assays for functional E3 ligase cascades, we have made this target class readily accessible for research and drug discovery.

Advantages of Ubiquitin Profiling with Eurofins:

• Choose from >20 biological pathways and >35 active complexes and enzymes
  
• Unmatched quality and experience as the first commercially available service for screening E3 cascades
  
• Service can either be performed in single point concentration or 10 point dose response curves for IC50 determination with our UbiquitinProfiler™ or UbiquitinProfilerIC50™ services
  
• Biologically relevant substrates used
  

Profile functional E3 ligase cascades

While ubiquitination offers an exciting new avenue to approach many difficult-to-treat diseases, there are certain aspects of ubiquitination that have provided an obstacle to drug development. The first complexity is that, unlike other post-translational modifications (e.g., phosphorylation), ubiquitination relies on several interconnected steps involving multiple enzymes (i.e., E1, E2 and E3). Secondly, while E3 ligases are the most attractive targets because of their potential specificity, they are the most complex - sometimes consisting of multiple protein constituents. Furthermore, many of the substrates themselves need to be post-translationally modified before they are recognized as substrates for ubiquitination.

Eurofins overcomes the restrictions to ubiquitination drug discovery. By applying our protein production and assay expertise, we've developed a suite of assays to measure distinct ubiquitination cascades. Each assay contains the complete complement of ubiquitin enzymes (E1 activating enzyme, E2 conjugating enzyme and E3 ligase) and a biologically-relevant substrate with appropriate post-translational modifications. The incorporation of ubiquitin into the substrate is quantitatively detected by electrochemiluminescence, which allows for the identification of inhibitors and activators.

Advantages of UbiquitinProfiler™:

• Assays for over 25 E3 ligase cascades to screen and profile inhibitors and enhancers
  
• Cascades consists of E1, E2 and E3 enzymes and substrates with the ability to deconvolute hits against each enzyme component
  
• Measure substrate ubiquitination with activity assays
  
• Electrochemiluminescence always for sensitive detection of ubiquitin modifications
  
• Run by the same experts who have run KinaseProfiler™ services for 15+ years
  
• Single point or full 10-point IC50 curves
  
• Data reports available in 4 weeks or less
  

Deubiquitination (DUB profiling)

Analogous to phosphatases' role in reversing phosphorylation of proteins, the deconjugating enzyme (DCE) proteases, including deubiquitinating enzymes (DUBs), similarly reverse the ubiquitin modification of proteins. By removing ubiquitin from target proteins, DUBs act to oppose the effects of ubiquitination on protein function or targeted destruction by the proteasome.

Advantages of DUB profiling with Eurofins:

• Key DUBs available, including those relevant for epigenetic modification
  
• LanthaScreen® for high assay robustness and reduction of false positives by autofluorescent compounds

Proteases are a class of enzymes which perform proteolysis, the mechanism of digesting or cleaving long chain proteins into smaller fragments by severing the peptide bonds that hold them together. Trypsin, a serine protease secreted by the pancreas, is commonly used in cell tissue culture to cleave the bonds that cells use to adhere to a flask.

Proteases also play a major role in disease. HIV-1 protease is essential to the viral replication process. MMP-9, a matrix metallopeptidase, plays a role in angiogenesis and is a therapeutic target for cancer. Because of their significance in the pathology of disease, proteases are a relevant drug target class.

Our broad portfolio of over 50 proteases spanning across the serine, cysteine, aspartic, and metalloprotease families was designed with you in mind to facilitate your drug discovery process and provide you with the answers you need.

Over 50 targets to support your research, including these families and targets:

• Families
  
  • Metalloprotease
    
  • Caspase
    
  • Cathepsin
    
• Targets of interest
  
  • HIV-1 protease
    
  • Proteasome
    
  • DDP4...and many other peptidases

Cyclic nucleotide phosphodiesterases (PDEs) are ubiquitously distributed throughout mammalian tissues and play a major role in cell signaling by hydrolyzing cAMP and/or cGMP. Due to their diversity and specific distribution at the cellular and subcellular levels, PDEs are able to selectively regulate various cellular functions. This regulatory ability implicates PDEs in various therapeutic areas including CNS, inflammation, and oncology.

As PDEs are expressed in a variety of tissues, selectivity is a prerequisite for a therapeutically viable PDE inhibitor. For example, high selectivity for PDE5 inhibitors is important for treating erectile dysfunction while minimizing possible side effects associated with inhibition of other PDEs. These possible side effects include tachycardia and vasodilation that are attributed to inhibition of PDE1 and PDE3, or blue-green vision disturbances that are attributed to inhibition of PDE6.

In order to meet the growing need for robust PDE profiling, Eurofins, through the expertise of Cerep and Panlabs, has designed an HTS platform to determine the inhibitory effects and selectivity of compounds on the PDE superfamily.

Advantages of PDE Profiling with Eurofins:

• Robustness and reproducibility confirmed by screening a broad panel of commercial available reference inhibitors and known commercial drugs
  
• Radiometric assays for high quality enzymatic assays
  
• Choose from over 20 PDEs, covering PDE1-11

Nuclear hormone receptors (NHRs) are a large superfamily of ligand-activated transcription factors that consist of two subtypes, steroid and non-steroid. These receptors serve as on-off switches for genes which regulate cell differentiation, proliferation, and metabolism, and thus can play a role in the pathology of cancer, cardiovascular disease, inflammation, and reproduction. As with many receptors, ligand binding induces a conformational change in the receptor, but unique to nuclear receptors is their ability to up- or down-regulate gene expression. This ability makes nuclear receptors interesting therapeutic and liability targets.

Eurofins offers a broad portfolio of both binding and functional assays, including co-activator recruitment for nuclear receptors. These assays can help provide a complete profile of your drug or compound and give you a better sense of its gene-regulating features.

Advantages of nuclear receptor profiling:

• Screening of a broad range of nuclear receptors
  
• Information on an important family of receptors responsible for a wide range of on- and
  off-target biological processes

Profile with the Industry's Largest Epigenetics Portfolio

Epigenetics, an increasingly popular field of drug discovery, is the study of heritable changes in gene expression that occur without changes in the DNA sequence. The epigenome is considered to be in constant flux and is responsive to environmental and pharmaceutically active molecules. Epigenetics-related enzymes and proteins catalyze the addition or removal of an array of covalent modifications on histone and non-histone proteins. For these reasons, epigenetics is becoming an increasingly important realm for drug therapies. Indeed, several drugs directed towards epigenetic modifying enzymes are already on the market with more in the clinic to treat cancer.

In 2012, ahead of all other CROs, Eurofins recognized the industry's upcoming need for a comprehensive epigenetics program containing all major families. To meet this growing need, we began to develop a range of target based assays using biologically relevant substrates (nucleosomes, histones, and peptides) to create the most meaningful assays. The majority of our proteins/substrates are produced in-house to ensure the highest quality of proteins resulting in high batch to batch reproducibility.

Eurofins' epigenetic toolbox is based on several technology platforms and allows for your choice of technology and/or substrates, providing added flexibility to meet all of your drug discovery needs.

Advantages of Eurofins' Epigenetics Program:

• Largest target and assay list in the industry, with over 160 epigenetic assays to choose from:
  
  • >50 Writers, including: SMMT, Kinases, DNMT, HMT and HAT
    
  • >40 Erasers, including: Demethylases, HDAC / sirtuin and DUB
    
  • >45 Readers, including: Bromodomain and Methyl readers
    
  • >17 Off-target and cell-based assays, including acetylation and methylation studies
    
• Compatible with screening in medium- or high-throughput
  
• All assays set up according to NIH guidelines
  
  • Employing biologically-relevant co-factors and substrates
    
  • Assay optimization includes apparent Km identification for assay constituents and assay linearization
    
• Assays can be modified to meet client needs
  
• Several technology platforms cover the epigenetic portfolio:
  
  • Gold standard filter-based radioactivity
    
  • RapidFire Mass Spectrometry
    
  • AlphaScreen
    
  • TR-FRET
    
• Customer tailored assays to supplement our off-the-shelf assay portfolio
  
  • A specific protein can be cloned, expressed, purified and have a corresponding enzymatic or binding assay developed using a selected technology platform

Eurofins high quality and well characterized PrecisION® ion channel stable cell lines are the foundation of IonChannelProfiler™ services. From single-cell manual patch clamp, the gold standard in ion channel work, to high throughput 384-well automated electrophysiology instruments, we use our cells on a variety of platforms to provide the throughput or depth of analysis that matches your needs.

Advantages of Ion Channel Profiling with Eurofins:

• Cell-based functional assays for more than 60 ion channels, one of the largest ion channel portfolios in the industry, representing:
  
  • Voltage-gated channels (e.g., sodium, potassium and calcium channels)
    
  • Ligand-gated channels (e.g., GABA, nAChR and TRP channels)
    
• Focus on electrophysiology to avoid missed detection related to other methods, allowing identification of:
  
  • Agonists, antagonists, allosteric modulators
    
  • State (closed, open and inactive) and use-dependent blockers
    
• Gold standard manual patch clamp assays for detailed characterization
  
• Variety of automated platforms to suit a range of throughput, including IonWorks® Quattro™, PatchXpress®, QPatch® - and the only CRO offering services on IonFlux HT™
  
• Assays are designed to work in up to 0.5% DMSO to minimize vehicle response and maximize result consistency
  
• hERGProfiler™ and CardiacProfiler™ services to address cardiac liabilities
  
• Quick turnaround times of 1-3 weeks depending on project types
  
  • Ongoing SAR-driven projects: approximately one week
    
  • Fixed schedule for CardiacProfiler™ screens: approximately three weeks

Multi-prong Assessment of Cardiac Liability

Cardiotoxicity is a major cause of drug development failure and withdrawals. hERG screening alone cannot reliably detect potential cardiac adverse side effects. The Cardiac Safety Research Consortium (CSRC) recently proposed the Comprehensive in vitro Proarrhythmia Assay (CiPA), which would incorporate in vitro cardiomyocyte studies as a component of cardiac safety assessment. In addition to our CardiacProfiler™ services, our services using human cardiomyocytes fit into this emerging paradigm.

We have employed a combination of assay platforms along with Cytiva™ Plus human stem cell derived cardiomyocytes from GE Healthcare to address this need for more biologically relevant and predictive cardiac liability models. Eurofins offers complimentary approaches, including High Content Analsysis (HCA), Multiple Electrode Array (MEA) and manual patch clamp assays, to reliably detect and characterize cardiac safety risks.

Using a population of beating cardiomyocytes, MEA provides ECG-like field potentials to detect proarrhythmic events, alterations in beat rate and other adverse effects. Changes in field potential duration is related to alterations of the QT interval, which in a clinical setting may initiate a potentially fatal Torsades de Pointes (TdP) arrhythmia.

Manual patch clamp studies provide a detailed assessment of action potential duration on individual cardiomyocytes. Specifically, ventricular-type cells within the general cardiomyocyte population can be tested. This is important as the ventricular-type cardiomyocytes are relevant for ventricular dysfunction, a major form of cardiotoxicity.

Imaging techniques using cardiomyocytes measure cell health parameters resulting from structural damage upon prolonged compound treatment, toxic effects that may not be otherwise identified by electrophysiological methods. Multiple parameters, including cell number, intracellular calcium flux, mitochondrial membrane potential and membrane permeability can be measured on live cells to determine changes in normal cellular functions. Alternatively, fixed cells are used to reveal appearance of peri-nuclear BNP expression, α-actinin disruption, troponin I integrity and reduction in cell number to further demonstrate toxic effects.

Advantages of Cardiotoxicity Assessment with Eurofins:

• Biologically relevant, highly characterized and reproducible cardiac cell model:
  
  • Differentiated human stem cells provide a reliable and reproducible source of cardiomyocytes
    
  • Highly pure cell population with cardiac marker expression (α-actinin, troponin I)
    
  • Large fraction of ventricular subtype cells to test ventricular dysfunction - the predominant form of cardiotoxicity
    
  • Functionally verified by patch clamp for both action potential and individual ion channel currents
    

Beyond hERG for cardiac liability testing

Gain a broader insight into potential cardiac risk with earlier profiling of new chemical entities at throughputs that are more appropriate for lead discovery and optimization. CardiacProfiler™ is a comprehensive cardiac safety panel that includes each of the key cardiac channels, providing a robust, cost-effective approach to cardiac liability testing.

Many withdrawn drugs have been shown to block the human ether-a-go-go (hERG) channel, delaying repolarization of the cardiac action potential and leading to prolongation of the QT interval on an electrocardiogram. This can potentially initiate the arrhythmia known as Torsades de Pointes (TdP) with fatal consequences. While hERG is an important factor for cardiac liability, it is not the only factor. Excitation and relaxation of cardiac muscle is regulated by a variety of different ion channels. Some of these channels are genetically linked with long QT syndrome, suggesting that their modulation by drugs could also produce life-threatening arrhythmias independently of hERG. Conversely, a number of relatively potent hERG channel blockers do not induce TdP. Many of these drugs (e.g. verapamil, amiodarone) have mixed channel blocking action which may ameliorate the effects of hERG blockade. Thus, hERG screening alone may be insufficient to flag potential cardiac liability. Conversely, abandoning promising compounds based on hERG blockade alone may result in failing potentially safe clinical drugs.

In support of this philosophy, a recently output of the Cardiac Safety Research Consortium (CSRC) was a proposed shift in cardiac safety assessment to include a Comprehensive in vitro Proarrhythmia Assay (CiPA). CardiacProfiler™ services as well as our related cardiotoxicity assays using human cardiomyocytes fit into this emerging paradigm.

Advantages of Cardiac Risk Profiling with Eurofins:

• Suite of cardiac ion channels with relevant association with Long QT syndrome
  
• Only provider licensed to offer an Cav1.2 L-type calcium channel
  
• PrecisION® cell lines recombinantly expressing human channels ensure meaningful high-quality, reproducible results you can trust
  
• CardiacProfiler™ service offered on a fixed schedule every three weeks

Transporter studies for ADME and therapeutic activity
Transporters are membrane proteins that control the influx and efflux of essential nutrients, ions, neurotransmitters, cellular waste, environmental toxins, and other xenobiotics such as drugs. Drug transporters are composed of two major super families: ABC (ATP binding cassette) and SLC (solute carrier) transporters.

ABC transporters mediate unidirectional efflux and are primarily active transporters as they directly rely on ATP hydrolysis to pump their substrates across membranes. They play major roles in hepatobiliary and urinary excretion of drugs from the blood to the lumen, intestinal absorption of drugs, and in brain blood barrier penetration of drugs.

SLC transporters mediate either drug uptake or efflux and can be facilitated transporters or secondary active transporters, which transport drugs using energy produced by transmembrane movement of other solutes in accordance with their concentration gradient. They play major roles in hepatic and renal uptake, and urinary excretion. Some SLC transporters, such as those for neurotransmitters, are also important drug targets themselves with clinical drugs treating various psychiatric disorders.

Transporters play a significant role in a drug's clinical efficacy and safety. Eurofins, through the expertise of Cerep and Panlabs, has developed robust transporter uptake and inhibition assays to allow quick and cost-effective screening of new molecular entities (NMEs) for their interactions with these transporters.

Advantages of transporter screening with Eurofins:

• Cell based assays to provide functional characterization of compound activity
  
• Key drug transporters and neurotransmitter transporters are covered
  
• Drug transporter assays meet regulatory requirements
  

Transporters for drug absorption and disposition

Drug transporters are essential in controlling the absorption and disposition of xenobiotics, including pharmaceuticals. As such, transporters play a key role in defining the clinical efficacy and safety of drugs.

ABC (ATP binding cassette) transporters, including members like P-gp and BCRP transporters, play major roles in hepatobiliary and urinary excretion of drugs from the blood to the lumen, intestinal absorption of drugs, and in brain blood barrier penetration of drugs. SLC (solute carrier) transporters, including members like OATP1B1 and OAT1, play major roles in hepatic and renal uptake, and urinary excretion.

Interaction with these efflux or uptake transporters can play a significant role in a drug's pharmacokinetic properties and, in turn, its efficacy and safety profile. Eurofins, through the expertise of Cerep and Panlabs, has developed robust in vitro transporter inhibition assays to assess potential drug-drug interactions prior to costly clinical assessment.

Advantages of Drug Transporter Screening:

• Assays meet regulatory requirements
  
• Services available for drug transporters of primary interest:
  
  • ABC transporters: MDR1/P-gp, BCRP,
    
  • SLC transporters: OAT1, OAT3, OATP1B1, OATP1B3, OCT2
    
• Additional drug transporter are available to provide a comprehensive profile of drug transport
  
• Recombinant cell lines developed internally for superior data reproducibility
  

Screening for therapeutic activity or selectivity profiling off-target liability

Neurotransmitter transporters, especially biogenic amine transporters, have been a highly successful target class for clinical therapeutics. First generation Selective Serotonin Re-Uptake inhibitors (SSRIs) are one of the most recognized class of drugs to treat depression. While next generation anti-depressants with polypharmacology (activity across multiple targets) for serotonin and norepinephrine may offer better clinical activity.

Like most drug targets, unwanted interaction with neurotransmitter transporters may lead to adverse reactions. Therefore, profiling compound activity against neurotransmitter transporters is also important for general liability assessment.

Eurofins, through the expertise of Cerep and Panlabs, have both binding and cellular uptake assays to test compounds for interaction against this class of clinically important targets.

Advantages of neurotransmitter transporter screening with Eurofins:

• Combination of both binding and cellular uptake assays to provide complementary assessment of compound interaction and function
  
• Coverage of important targets, including:
  
  • Serotonin
    
  • Norepinephrine
    
  • Dopamine
    
  • Adenosine…and more

In additional to playing play a major role in the metabolic fate of drugs, cytochrome P450 (CYP) enzymes are major targets in drug-drug interactions (DDIs). Drug induced inhibition of CYP enzymatic activity is one important cause for DDI, but it not alone sufficient to assess the potential to induce DDI. CYP enzymes can be transcriptionally induced by many xenobiotics including some drugs, through the mediation of nuclear receptors. Therefore, understanding the potential of new molecular entities (NMEs) to induce CYP expression is important to minimize DDI liability.

CYP induction is traditionally determined by measuring enzymatic activity in cultured primary hepatocytes; however, due to enzyme inhibition, cytotoxicity, or other effects, this assay is prone to false negatives. In order to address the growing need for robust, sensitive assays to measure CYP induction, Eurofins has developed CYP assays which measure the mRNA level of the targeted CYP genes. As the preferred method to measure CYP induction, mRNA analysis is less prone to false negatives than traditional activity-based assays and follows FDA and EMA Guidance (2012).

Advantages of CYP induction profiling with Eurofins:

• mRNA measurements for CYP isozymes (CYP1A2, CYP2B6 and CYP3A4) induction provide a more sensitive and reliable method than traditional enzyme activity assays
  
• Combine with other drug-drug interaction studies to gain a complete profile of your new molecular entity
  
• Assays established in accordance with the FDA Guidance on Drug Interaction Studies (2012)1
  
  • Cryopreserved human hepatocytes from at least three donors
    
  • Real time qPCR
    
  • Results can be used for regulatory filings

Ion channels are well known for regulating electrical activity in excitable cells, and many roles in non-excitable tissues continue to be uncovered. They are important therapeutic targets in a range of indications including arrhythmia, hypertension, local anesthesia, pain, stroke, epilepsy, depression, bipolar disorder, COPD, autoimmune disorders and diabetes. Not only are ion channels important drug targets, but they are also generally relevant for drug safety. Indeed, many drugs withdrawn from the market due to cardiac related adverse effects have been shown to block the human ether-a-go-go (hERG) ion channel, which delays repolarization of the cardiac action potential and can result in a potentially fatal arrhythmia known as Torsades de Pointes (TdP).

Through the combined expertise of Cerep, Panlabs, and IonChannelProfiler™ services, Eurofins offers a broad portfolio of both binding and functional assays for ion channels. Our years of experience as an ion channel CRO has enabled us to better understand our customers' latent needs, so if you're looking for an ion channel screening solution, you've found it.

Advantages of Ion Channel Profiling with Eurofins:

• Robust binding and functional cell based assays performed with stability-tested PrecisION® cell lines
  
• Assays for agonist, antagonist, NAM and PAM activity
  
• Over 9 years of experience and the first CRO to provided testing by automated electrophysiology in 2005
  
• Only CRO in the industry that offers services on IonFlux HT™
  
• Over 60 ion channel targets and >130 assays available on Manual Patch Clamp, IonWorks®, PatchXpress®, IonFlux HT™ QPatch® systems
  
• Quick turnaround times of 2-3 weeks depending on project type

Early & Late Panel Assays

Providing critical early assessment across a range of targets, our safety and liability service panels use a combination of functional and binding assays to empower you to make better decisions. When you know everything you can about your drug candidates, including potential off-target liabilities, you'll avoid costly setbacks and develop more promising compounds, faster.

Each of our safety and liabilities panels were specifically chosen for their relevance to critical diseases and biological processes, including neurotoxicity, cytotoxicity, cardiac function, immunoprotection, diabetes, inflammation, and gastrointestinal functions. These panels can provide early identification of potential adverse activity in order to help you optimize your drug's safety margins.

Profiles matched to the stage of your project:

• Hit to lead panels
  
  • Designed to aid in selection of potential lead compounds from hits
    
  • Small number of assays to identify general selectivity and liability issues
    
• Lead optimization panels
  
  • Greater number of assays
    
  • Balance between targets with known safety liabilities and those providing information on selectivity
    
• Safety panels
  
  • Oriented more toward targets with known safety liabilities than toward providing information on selectivity
    
    • SafetyScreen44™ - 44 targets, recommended by four major pharmaceutical companies
      
    • CardiacProfiler™ - functional testing against eight Cardiac ion channels, including hERG, Nav1.5 and Cav1.2
      
• Pre-IND Panels
  
  • Complete picture of the off-target activities of drug candidates before IND filing
    
    • DiversityPanel - a broad array of targets to provide a comprehensive picture of potential adverse effects
      
    • Bioavailability panel - provides a thorough assessment of compound bioavailability and potential drug-drug interactions

G protein coupled receptors (GPCRs), also known as seven transmembrane receptors (7TMs), have been and will continue to be prominent drug targets for treating hypertension, pain, asthma, neurological and other disorders. Indeed, as 40% of pharmaceuticals target a small fraction of the family, GPCR profiling continues to maintain an important role in the realm of drug discovery and development.

Eurofins has all the solutions for your receptor screening and GPCR profiling needs through the combined expertise of Cerep, Panlabs and GPCRProfiler® services . Whether you're looking for binding assays, functional profiling or secondary messengers, we have you covered. If you're embarking on a new project and want to outsource an HTS campaign, SAR or general selectivity profiling of your leads and candidates, we have a receptor profiling solution for you.

Advantages of GPCR profiling with Eurofins:

• Robust binding and functional, cell based secondary messenger assays to suit all your research needs
  
• Assays for affinity, agonist, antagonist, inverse agonist and positive allosteric activity
  
• Coverage of over 165 targets with >590 of assays
  
• 20 years of experience as a GPCR CRO
  
• Quick turnaround times of 1-3 weeks depending on project type
  

Radioligand Binding Services

Radioligand binding is the gold standard method for detecting compound interaction with GPCRs, allowing for the simplest interpretation of assay results. A favorite of medicinal chemists, radioligand displacement assays can be used for simple yes/no interaction determination or to calculate a compound's affinity for a receptor of interest. Along with functional assays, binding studies can provide a complete profile of a compound's in vitro pharmacology.

Eurofins is the market leader for outsourced radioligand binding studies. With origins in Cerep and Panlabs, we have been successfully conducting GPCR binding studies for decades. Our binding assays use the gold standard filtration method for highest assay robustness. The majority of our assays are conducted with membrane preparations produced in-house to control for the quality of these critical reagents and ensure data reproducibility.

Cell-based assays for GPCRs

When assessing a GPCR therapeutic's potential efficacy and liabilities, it is important to understand its on and off target activity. GPCRs can signal through a variety of cascades and a drug will activate, inhibit, or modulate one or more of these. Indeed, compound activity can be complex as certain compounds may elicit different activity on different signaling cascades, also known as ligand biased signaling. Eurofins has services covering several second messenger pathways to better understand compound activity. Along with binding assays, these functional assays can provide a complete profile of a compound's in vitro pharmacology.

GPCRProfiler® services is the first complete cell-based functional GPCR profiling platform that uses a common validated readout for over 165 GPCRs. The foundation of GPCRProfiler® is our ChemiScreen™ GPCR stable cell lines that are used for real-time calcium flux assays to rapidly, reliably, and reproducibly screen and profile compounds. Using one platform allows ligands to be screened for agonist and antagonist activity using identical buffer conditions and incubation times for the entire spectrum of GPCRs for easy high throughput analysis and comparison.

Our cell based assays feature multiple second messenger readouts, including cAMP for Gi and Gs coupled receptors as well as IP1 and IP3/calcium flux for Gq coupled receptors, emphasizing the natural pathway of the receptor. Additionally, we also have label free cell based assays to look at endogenous receptor signaling as well as traditional GTPγS assays.

Together, our collection of functional assays, along with our binding studies, can provide an orthogonal approach to confirm the in vitro pharmacology of your compounds.

GPCR Profiling for Allosteric Modulators

Historically, GPCR drug development has been focused on compounds that interact with the receptors' orthosteric site; however, the orthosteric binding site can be highly conserved, making it extremely difficult to generate receptor-specific compounds.

Researchers circumnavigate this restraint by exploiting the receptor's allosteric sites. Allosteric compounds are gaining relevance because they can modulate receptor activity, allowing greater physiological control of receptor activation, and are predicted to be more selective and have a reduced potential for overdose.

Our AllostericProfiler™ receptor profiling services use a functional readout to detect allosteric compound activity. AllostericProfiler™ is the first fully validated selectivity profiling service capable of detecting a range of compound activities for over 165 GPCRs by using a unique two-addition methodology to detect of a wide variety of activities including agonist, positive allosteric modulator (PAM) and negative allosteric modulator (NAM) activity.

AllostericScreener™ screens compound libraries to identify new compounds having positive allosteric modulator (PAM) activity for a target of interest. AllostericScreener™ service employs an assay design that is similar, but slightly modified, to our standard GPCRProfiler® services. This assay design can identify both agonist and PAM activities. Therefore, you can discriminate hits that either activate the receptor directly, modulate the receptor's activity or those that have a mixture of these activities.

Drug discovery efforts are usually centered on a single target against which compounds are optimized for therapeutic efficacy. However, developing a selective kinase inhibitor is particularly challenging, because of conserved active sites and high structural homology between kinases. Broad profiling, for many, has become a requirement of new kinase inhibitor programs for better understanding structure/activity relationships, helping to avoid targets with toxic liabilities, and, ultimately, to select better drug candidates. The last five years have seen eight anti-cancer kinase inhibitors receiving FDA approval, increasing recognition of kinases as a vital drug target class.

Through the combined expertise of Cerep, Panlabs, KinaseProfiler™, and SignalProfiler™ services, Eurofins offers an unmatched portfolio of biochemical and cellular kinase assays.

Advantages of KinaseProfiler™ services:

• One week guaranteed turnaround time to drive SAR*
  
• Choose from over 600 radiometric and TR-FRET (HTRF® or LANCE®) kinase activity assays, one of the largest panels in the industry
  
• Unparalleled quality control
  
• Low variation across replicates (including controls)
  
• Selection of ATP concentrations, including Km and 10 µM
  
• Single point or full 10-point IC50 curves
  
• HTRF® lipid PI3 Kinase assays
  
• Flexibility to include non-standard solvents or pre-incubation
  
• Reserve assay slots to allow a steady flow of compounds for the fastest possible throughput
  

Benefits of profiling cellular kinase activity with Eurofins:

• Multiplexed analysis of key pathways including:
  
  • Receptor tyrosine kinase (RTK) pathways
    
  • MAPK pathway
    
  • Akt/mTOR pathway
    
  • p38/SAPK pathway
    
  • Src kinase pathway
    
• Inspection of common intracellular signal transduction cascades and networks
  
• Can detect inhibition of either the ligand binding domain or kinase domain
  
• Cellular kinase phosphorylation and tyrosine kinase receptor activity assays
  
  • >70 Singleplex and >20 Multiplex assays
    
  • MILLIPLEX MAPmates™ or AlphaScreen® Surefire® technologies for either multiplexed or higher throughput assays
    
• Single-dose or EC50/IC50 cell-based determinations

Eurofins has the experience and skills needed to bring you early stage pharmacokinetic (PK), oral bioavailability and blood-brain barrier (BBB) data to evaluate your new molecular entity. PK and BBB studies are prerequisite for interpreting preclinical efficacy and toxicology results. Using qualitative measurements of drug exposure, we can provide a solid interpretation of preclinical efficacy. PK data can also help in species selection and design of preclinical toxicology studies.

Eurofins will work with you to understand the physicochemical properties of your test material to develop a dosing and sample collection strategy that will provide you with the most meaningful data. We can obtain blood and/or tissue (e.g. brain) samples following dose administration (IV, PO, IP, IM, SC, IN, and ICV, also short- or long-term continuous infusions) and both serial sampling for PK and parallel sampling for PK, BBB or combined PK-BBB are available. PK/PD studies can be combined using our broad portfolio of pharmacodynamics models.

PK/BBB dosing and sampling studies can be paired with standard or custom bioanalytical methods to provide a concentration versus time curves to provide a complete pharmacokinetic profile for your compound.

Advantages of in vivo pharmacokinetic and blood/brain barrier studies with Eurofins:

• Extensive experience with multiple routes of administration, including long-term continuous infusion
  
• Proficient in performing PK and BBB studies
  
• Combined PK/BBB sampling and bioanalytical studies
  
• Renal and biliary clearance
  
• Experience you can trust: >25 years running in vivo preclinical services with a staff averaging 15 years of experience
  
• Long range of efficacy and toxicity studies can be conducted at the same provider

Cardiovascular disease (CVD) is one of the leading causes of death for both men and women worldwide. CVDs affect the heart and blood vessels, leading to a variety of conditions including ischemia, stroke, heart attack, angina and cardiomyopathy. Eurofins, through the expertise of Panlabs, has been conducting in vivo testing for cardiovascular disease using in vivo models for both side-effect profiling and efficacy for over 25 years.

Our staff, with their vast experience, is dedicated in assisting you to properly evaluate your lead series and clinical candidates. We offer a number of relevant cardiovascular disease models, including models of hypertension, thrombosis and arrhythmia.. We can also provide many related ancillary services such as measuring relevant cytokine biomarkers in plasma though our highly sensitive ImmunoSignal™ services, measure lipids or perform blood chemistries.

Whether you are trying to evaluate potential cardiovascular liabilities during lead optimization or trying to verify in vivo efficacy, Eurofins can provide you high quality and reproducible results to progress your drug discovery program.

Advantages of Eurofins' cardiovascular in vivo studies:

• Over 30 cardiovascular disease/dysfunction assays available
  
  • Hypertension
    
  • Ischemia-reperfusion injury
    
  • Dyslipidemia
    
  • Arrhythmia and QT interval
    
  • Thrombosis
    
  • Adrenergic mediators
    
  • Clotting homeostasis
    
  • General cardiovascular safety assessment
    
• Experienced in combination studies with standard of care control regimes
  
• Models are qualified in a dose-dependent manner with approved benchmark positive controls.
  
• Companion services can monitor blood and organ exposure and biomarker analysis for PK/PD assessment
  
• Ancillary services: clinical chemistry, lipid profiles, hematology and blood gases.

In addition to our expansive portfolio of functional ion channel assays, we offer radioligand binding assays across a number of ion channel families including sodium, potassium, and calcium. Radioligand displacement assays can be used for a yes/no interaction determination or to calculate a compound's affinity for a channel of interest. Along with functional assays, binding studies can provide an orthogonal approach to creating a complete profile of your compound's in vitro pharmacology.

Our binding assays use the gold standard filtration method to produce highest assay robustness. Our ion channel binding assays are conducted with membrane preparations produced in-house to control for the quality of these critical reagents and to ensure tight data reproducibility. Radioligand binding assays are a useful approach for monitoring potential hERG liability at the earliest phase of drug discovery. Here, the primary aims are to raise flags which could trigger follow-up studies, such as electrophysiological screening.

Advantages of Ion Channel Binding with Eurofins:

• Gold standard filtration assays
  
• Choose from sodium, potassium, or calcium channels
  
• Majority of membrane preps produced in-house for guaranteed lot-to-lot consistency
  
• Capacity: >10 scintillation counters
  
• Quick turnaround times of 2-3 weeks depending on project types
  
• hERG membrane preparations (CYL4039) also available for purchase for in-house counter-screening

Eurofins Villapharma provides the biopharmaceutical industry with high quality organic synthesis expertise and unique collections of novel, patentable, drug-like organic compounds for drug discovery. Our teams, of more than 80 chemists, are experts in high-quality Organic Synthesis, Medicinal Chemistry Services, Screening Libraries, and the production of unique collections of novel organic molecules with potential biological activity. Our proven experience in developing Screening Libraries has resulted in new chemical starting points for multiple client discovery programs, including those in the Anti-infective, Oncology, CNS and Metabolic Disease therapeutic areas.

With our state of the art facilities, a production technology that is based on rapid parallel synthesis by using specially designed versatile workstations, and a large and interesting portfolio of privileged core structures, our platform allows us the ability to cover a wide range of different chemistries. The Eurofins Villapharma chemists leverage their high level of problem solving skills to manage programs needing complex chemistry and our chemists are recognized for the strong expertise they have developed in heterocycles, macrocycles, peptides/peptidomimetics, and natural product-like chemistry.

Our team follows established processes and Standard Operating Procedures (SOP’s), and has the necessary tools required for optimal data and project management. Furthermore, the team has a proven track record of handling confidential data and exchanging of files through secure file sharing systems. Data and records are supported through a proprietary Electronic Lab Notebook system in order to ensure reproducibility, quality control, and management of intellectual property.

Pharmacology Services