Services

Selvita offers custom assay development and screening/profiling services up to 1536-well plate format.  We have created a robust compound management infrastructure supporting manipulation of samples in racks, tubes and plates.  We employ a fully automated HTS platform allowing testing of 10,000 compounds a day across a wide range of biochemical and cellular assay types.  We operate HCS screening in support of client projects within our new state of the art HCS laboratory and are generating novel machine-learning based scripts to support enhanced image analysis in the cell types of particular interest to our clients.

Selvita has initiated the design and building of a new HTS library which will be grown and developed further to provide an excellent resource for hit identification. This compound collection is supplemented by the FideltaMacro™ library with over 1300 diverse macrocyclic molecules.

Our team is built on extensive expertise in the areas of molecular biology, biochemistry, immunology and cell biology. We have a track of record in assay development for various classes of targets, including enzymes (kinases, phosphatases, metabolic enzymes, immuno-modulatory enzymes) and protein receptors (GPCRs, nuclear receptors). Selvita’s state of art-equipment allows for a wide variety of detection modes to be implemented, adding value to your drug discovery program.

Biochemical and cell-based assays

Supported readouts:

• Absorbance and Fluorescence (endpoint, kinetic)
• Fluorescence Resonance Energy Transfer (FRET) and Time -Resolved FRET (TR FRET)
• Fluorescence Polarization (FP)
• Luminescence
• Scintillation counting (3H and 14C)
• Cell based reporter & viability assays
• AlphaScreen bead-based proximity assays

Biophysical assays

• Surface plasmon resonance (SPR)
• Thermal shift (differential scanning fluorimetry)
• Microscale thermophoresis (MST)
• Isothermal titration calorimetry (ITC)
• Protein-ligand NMR

Selvita can offer case-by-case selection of suitable cellular model/s to investigate the biological mechanism of interest (e.g. cancer cell panels of a specific type and/or with common mutation patterns). Our in-house capabilities for reagent generation include in-house production and validation of recombinant cell lines and recombinant proteins. Our assay development strategies are characterized by efficient and effective protocols. Usually, multi-factor design of experiments is employed during method development.  It accelerates optimization of the assay performance, thus helping to obtain a measurement system that meets its requirements. Statistical parameters are used to support a reliable and robust assay for routine use, e.g. IC₅₀/EC₅₀; mRSD (mean Relative Standard Deviation); S/N ratio; Z’ factor; plate uniformity.

Medicinal chemistry is an interdisciplinary science which plays a crucial role in the drug discovery process. In medicinal chemistry we follow a highly data-and-hypothesis driven path making meaningful molecules whilst also using productive synthetic routes to bring benefit both from efficiency and from serendipitous discoveries.

We are skilled at multi-parameter optimisation based on the analysis of biological, structural and physicochemical data. We use these expanding data sets from ADME assessments, computer-aided drug design, structural biology, in vitro and in vivo biological testing, and disease models to refine our thinking and to influence the design of new iterations of molecules. In addition, the effectiveness with which the projects are executed is very dependent on the quality of the infrastructure but even more by the quality and passion of our people.

We benefit from close interactions with colleagues in the associated scientific disciplines and execute our work together under one roof in our drug discovery facilities in Krakow. Our skill in chemistry is built on years of diverse experience in small molecule synthesis and incorporates strengths in the efficient preparation of compounds across multiple chemotypes including heterocyclic synthesis, macrocyclics, peptidic molecules, steroids and nucleosides, and beyond. Our strong asset is “out-of-box” thinking and application of the latest techniques like flow-, electro- and photochemistry where classical methods reached their boundaries.

The biology offering at Selvita covers all drug discovery purposes from target validation through to preclinical candidate selection. We provide a protein-to cell approach for the development of pharmacologically active compounds.

Our in vitro pharmacology activities commence from the customized production of recombinant proteins for use as screening proteins to probe compound interaction in a panel of biochemical and biophysical methods, also in a high-throughput screening mode. In parallel, using Selvita’s considerable internal experience of protein crystallization, protein structural analyses can be performed by X-ray. Using cell-based assays, we perform small molecule activity analysis using of a large repository of cells.

These cell-based assays can also be run in a high-throughput screening mode for hit finding. Close to two hundred recombinant cell lines are available for assay development, adaptable to many pathological areas. Selvita offers the custom production of recombinant cell lines expressing the protein target of interest. In addition, primary cells, for example from human (blood) or from rodent (CNS), are available upon request for assay development. Secondary screening for pharmacological evaluation of compound characteristics is part of our Integrated Drug Discovery expertise. Mode of action, biomarkers, target engagement studies are provided for hit characterization, and for lead finding and optimization.

Multiple readouts are available from classical fluorescence, interference (TR-FRET) or biochemical-based up to bioluminescence gene reporter assays or Alpha-Lisa technology, from medium throughput to HTS, and up to 1536 well plates depending on the scalability of the assays. In vivo evaluations can be arranged by Selvita during the lead optimisation and prior to preclinical development candidate selection. We analyse tissue samples, generate and interpret PK parameters or drug and biomarker concentration for pharmacodynamic studies in many disease areas such as cancer and CNS.

With numerous disease models developed in the past 10 years, and the experience built within the pharmaceutical industries, biotech and academia, our highly professional in vivo pharmacology team supports drug discovery projects, fully integrated with other preclinical disciplines.

The core areas of our PD expertise are inflammation, infection, fibrosis and we intend to expand into additional areas in 2021. We apply a comprehensive approach to primary and secondary pharmacodynamic profiling of the compounds to ensure generation of high-quality data relevant to human disease. Our in-depth expertise in translational biomarker selection and validation gives confidence in the smooth transition of the candidate molecule into the clinic.

Individual efficacy studies or tailored in vivo pharmacology packages can also be provided, to cover specific client needs. A full panel of automated clinical biochemistry, haematology and coagulation analyses as well as histopathology and a range of molecular biology techniques are available to support investigation of the compound efficacy.

Our toxicology group supports integrated drug discovery projects by standard and customized studies in rodents.

Our primary objective is to discharge in vivo safety liabilities through tailored study design. We offer you investigative, problem solving studies with fit-for-purpose, integrated assessment of multiple endpoints (general toxicology, pathology and DMPK endpoints).

You can benefit from our broad experience with different targets and mechanisms as it allows us to deal with unexpected findings or toxicity issues during the course of research projects.

In vivo experiments are performed at Fidelta, Selvita Group company, in AAALAC-I accredited State-of-the-Art animal facility (BSL2) with the competent veterinary personnel, technicians and hygiene personnel holding training certificates equivalent to FELASA categories A to D. Animal related research is based on the 3R principles and high-performance standards ensuring animal welfare, humane and ethical use of animals in scientific research.

Selvita offers a standard panel of in vitro ADME assays, which can be customized to fit client requirements, and provides comprehensive bioanalytical support based on our mass spectrometry platform. Our mission is to successfully support our Clients along the drug development pathway, to deliver high quality results provided by our experienced ADME specialists using state-of-the-art technologies. We provide fast turnaround times and maximum flexibility to deliver fit-for-purpose studies and achieve the specific goals of our client projects. ADME studies are offered either as stand-alone projects (fee for service model) or as a part of a collaborative Integrated Drug Discovery (IDD) program. In both cases, SLV provides a flexible approach by customizable protocols, which covers a very wide range of different needs. In addition, Selvita can propose a panel of ADME and Pharmacokinetic parameters analysis in a preassembled package which can be easily changed and customizable. The panel is ideally divided into a tiered approach depending on the drug discovery phases and then used for filtering and optimizing the ADME properties of compounds in a SAR fashion. Selvita will provide the full or partial ADME/PK package and also variants (e.g. urine analysis, tissues stability and others) following Client requests or project team decisions. Eventually, Selvita’s ADME/PK expert analysts can suggest the best compounds to be progressed into LO phase and in vivo PD experiments. Our Bioanalytical capabilities cover analyses of small molecules supporting both non-GLP as well as GLP- studies including a wide range of different activities from simple qualitative analyses to complex validation of analytical methods. Selvita also has broad experience with different sample preparation approaches using biological materials.

ADME tests:

Physico-Chemical Profiling

• Kinetic Solubility
• Thermodynamic Solubility
• Lipophilicity (LogD/LogP)
• pKa determination In vitro Permeability Studies Passive Transport Assessment

In-vitro Metabolism and Drug-Drug Interaction

• PAMPA GI • PAMPA BBB Active Transport Assessment • Caco-2 Permeability
• MDR1-MDCKII Permeability In vitro Metabolism Studies
• Microsomal stability – liver or intestinal microsomes and homogenates
• S9 Stability
• Hepatocyte Stability
• Aldehyde Oxidase reaction phenotyping
• Plasma, whole blood and biorelevant media stability
• Metabolite Profiling and Identification

In vitro Binding Studies:

• Plasma Protein Binding – Rapid Equilibrium Dialysis
• Plasma Protein Binding – High Sensitivity Binding Kit
• Tissue Binding – Rapid Equilibrium Dialysis CYP Studies
• Inhibition of the cytochrome P450 isoform activity (IC50, screening assay)

Bioanalytical Capabilities:

• Method Development/Transfer/Verification/Validation
• Qualitative Analysis
• Quantitative Analysis Matrices: Plasma and Tissue Homogenates

Department of Biochemistry has long-standing expertise in supporting development of biologically active chemical compounds for multiple pharmaceutical, agrochemical and biotech companies. Extensive know-how combined with highly skilled and goal-oriented scientists is a perfect combination enabling a successful execution of your project.

Our services are dedicated to assist in multiple steps during the process of active compound development by providing high quality recombinant proteins and comprehensive structural information. Flexible pipelines employed by Selvita ensure efficient delivery of tailored protein products for various research purposes including crystallographic, biophysical, enzymatic or in vivo studies. Our X-ray services are designed to provide high-resolution structural information to support hit validation, hit-to-lead development and lead optimization in the most time/cost efficient manner.

Protein Production

Solid expertise of our scientific team with a broad range of target proteins make a strong basis for meeting Client expectations. Our offer includes:

• coli, insect cell and mammalian expression systems
• Customized production of target proteins on a mgs to tens of mgs scale
• Development and optimization of purification protocol to maximize the protein quality (typically >90-95% purity)
• Tailored protein products (g. tags, non-radioactive isotopic labelling – ¹⁵N, ¹³C, low-endotoxin levels)
• Standard Quality Control evaluating protein concentration, purity, stability, homogeneity and oligomeric status
• Detailed reports

Macromolecular crystallography

Structural studies using X-ray crystallography are driven by a team with many years of international experience. Altogether, we have determined 200+ high-resolution structures and published 60+ research papers. Our services include:

• Screening of 1200+ crystallization conditions using a nanodispensing robotic system
• Optimization of initial hits, including, but not limited to, grid screening around the primary crystallization conditions and seeding approaches
• Secured access to synchrotron measurements across Europe
• X-ray data processing and analysis using XDS and CCP4 packages
• Structure determination using Molecular Replacement and Experimental Phasing
• Providing structures of protein-small molecule, protein-protein and protein-nucleic acid complexes
• Detailed reports

NMR in Drug Discovery

• NMR screening for FBDD
• NMR for ligand receptor interactions

Computer-Aided Drug Design Selvita’s computer-aided drug design (CADD) centre was created to foster collaborative research between medicinal chemists, biologists, biophysicists, structural biologists and computational scientists. In a drug discovery campaign, CADD is usually used for three major purposes:

1. Physico-chemical and structural characterization of compounds from various libraries followed by filtering large compound libraries into smaller sets of predicted active compounds that can be tested experimentally;
2. SAR analysis to guide the optimization of lead compounds, whether to increase its affinity or optimize drug metabolism and pharmacokinetic (DMPK) properties including absorption, distribution, metabolism, excretion, and the potential for toxicity (ADMET);
3. Design novel compounds with a help of a miscellaneous set of computational tools.

CADD depends on the extent of structure and other information available regarding the target (enzyme/receptor/protein) and the ligands. The latest advancements like AI, QSAR, combinatorial chemistry, different databases and available new software tools provide a basis for designing of ligands and inhibitors that require specificity and novelty.

At hit identification, hit-to-lead or lead optimization stage our CADD can apply the following methods:

• Ligand-Based Drug Design, LBDD (flexible shape-based and/or field-based alignments with active molecules when 3D structure of target is unknown, pharmacophore modelling)
• Structure-Based Drug Design, SBDD (ligand-receptor interaction studies with known 3D structure of receptor, docking, molecular dynamics)
• Homology modelling of proteins
• Chemoinformatics
• In silico physico-chemical profiling
• In silico ADMET profiling
• Focused libraries design for screening or synthesis (in silico or in collaboration with our medicinal chemists)
• HTS data analysis (hit confirmation and hit expansion to provide missed starting points for the project)
• SAR analysis and proposal of new compounds within currently investigated and/or new chemical series (bioisostere replacements, scaffold hopping)
• Quantitative Structure-Activity Relationship (QSAR) analysis
• Quantitative Structure-Property Relationship (QSPR) analysis
• Support to NMR screening for Fragment Based Drug Discovery (FBDD)

Most notably, our computational and medicinal chemists are working seamlessly together in interactive molecular design sessions, in order to rapidly generate hypotheses for molecular drug designs that are immediately assessed in suitable in silico models before molecules are prioritized and selected for syntheses in our laboratories or purchasing. Using a diverse set of available computational chemistry tools that are applicable for a given project, we assist the medical chemists in order to minimize the time, synthetic efforts and detours along the transformation process of a hit compound into a candidate.

AI-driven Drug Discovery

At Selvita not only we use traditional CADD but we also employ methods based on artificial intelligence (AI) or machine learning (ML). Those methods are especially useful for designing focused screening libraries, as a support in properties optimization process or to predict ADMET properties. Since the main task of the AI-based methods is to find associations between seemingly unrelated data, they are also very useful in target deconvolution process. All tasks related to AI and ML are performed in collaboration with ARDIGEN – SELVITA GROUP company specializing in this area.