Sygnature Discovery Ltd

Our medicinal chemistry department is one of the largest, most experienced and best equipped in the UK. Sygnature has an international team of pharmaceutical industry-experienced medicinal chemists (mainly PhD qualified bench scientists).

Our medicinal chemists have consistently created value for clients and delivered results which have accelerated drug discovery projects into pre-clinical development. FTE-based collaborations are regularly renewed and expanded amply demonstrating our clients’ confidence in our ability to deliver pivotal results.

Synthetic Chemistry

Sygnature's pharmaceutical industry-experienced medicinal chemists possess extensive synthetic chemistry capabilities and expertise. These skills are used during medicinal chemistry-driven projects, array/library preparation, when making chemical building blocks, or synthesising literature compounds.

• Multi-stage synthesis of complex molecules
  
• Chiral synthesis, chiral resolutions and chiral HPLC (analytical and semi-preparative)
  
• Synthesis of pro-drugs
  

• Enzyme-mediated chemistry experience

• Synthesis of small peptides and peptide mimetics

• Parallel synthesis of lead-seeking arrays or focused libraries

• Microwave assisted synthesis

• Hydrogenation (metered delivery and large scale)

• Flow chemistry capability

• Ozonolysis capability

• Photochemistry capability

• Scale-up syntheses using 5l and 12l flange-head vessels (up to 0.5 kg) with heating or cooling from -20 to +165c

Sygnature's Bioscience group is expert at developing tailored assays and building complete screening cascades. Assays can be developed 'from scratch,' established through collaboration with reagent providers, acquired commercially, or based on the client's existing assays and experience. Compounds can be assessed for potency, efficacy, kinetics, mechanism-of-action, selectivity, species crossover, etc. Additionally, automated liquid handling provides rapid and reliable turnaround of routine screening data, which is essential in efficiently driving successful integrated drug discovery projects.

In silico based vHTS offers an alternative, complementary and cost-effective approach to hit finding. Sygnature applies this technique to biological targets which have X-ray crystallographic data and/or known active compounds, or where a homology model can be generated. In silico screening of our ~8 million virtual compound library (see below) could therefore involve the docking of millions of compounds into the biological target (structure-based virtual screening), and/or searching for compounds similar to known active molecules (ligand-based virtual screening) using pharmacophores, ligand shape, electrostatic similarity and ligand fingerprinting strategies. Using our exclusive access to the HPC Facility and Amazon Web Services, vHTS projects involving millions of compounds can be undertaken in a matter of hours.

Lead Optimisation - Structure-Based Design

A number of approaches may be adopted during the lead optimisation phase of an integrated drug discovery project which incorporate structure-based design:

• Use of the fully-interactive ligand editor from Molsoft
  
• Visualisation of electron density maps
  
• Water assessment technology to target crystallographic waters in ligand design
  
• X-ray crystallography collaborations can be undertaken on behalf of customers
  
• Protein structure visualisation to communicate with medicinal chemistry teams
  

Lead Optimisation - Focused Array Design

Sygnature Discovery is able to use in silico approaches to design focused arrays of compounds during lead optimisation. SciFinder and other databases are interrogated to identify potential starting points of interest. These are then downloaded as SDF files and enumerated into a library focused on the biological target of interest. In silico filters are then applied based on desired physico-chemical properties, docking score, pharmacophore fit etc. Molecules are then elected for synthesis to deliver a focused array of compounds for subsequent in vitro testing at Sygnature.

The DMPK and Physical Sciences Department is dedicated to the understanding of the absorption, distribution, metabolism and excretion of drug candidates. Our industry experienced scientists work in close partnership with the client and other departments within Sygnature.

We deliver valuable and reproducible DMPK data using industry standard assay technologies and techniques that support an efficient design, make, test, analyse cycle during an integrated project.

Our scientists not only undertake the in vitro assays, but also have the expertise to interpret the data generated. They also have the know-how and experience to provide a comprehensive consultancy service to interpret in vitro DMPK and in vivo PK data generated elsewhere. In addition, throughout the course of a project our clients have direct access to our scientists to facilitate complete transparency and detailed discussions.

Examples

The following are examples of DMPK and Physical Sciences assays which typically form part of an integrated drug discovery project undertaken at Sygnature:

• Physico-chemical profiling
  
  • Solubility (kinetic), LogD (micro shake-flask), pKa, PAMPA, Chemical stability
    
• Absorption
  
  • Caco-2, MDCK
    
  • Transporter inhibition
    
• Distribution
  
  • Plasma protein binding (rapid equilibrium dialysis) (RED device)
    
  • Blood-plasma partitioning
    
  • CNS/Brain exposure
    
• Metabolism
  
  • Microsomal stability (Clint & t1/2)
    
  • Hepatocyte stability (Clint & t1/2)
    
  • Plasma stability (t1/2)
    
  • Metabolite profiling and identification (LC-MS-TOF based)
    
• Bioanalysis and Pharmacokinetics
  
  • Support PK studies (in-life phase outsourced)
    
  • PK/PD relationships and dose predictions
    
• In-house instrumentation
  
  • Waters Xevo G2-XS QTOF (UPLC-MS-TOF)
    
  • Waters Xevo TQS-micro (UPLC-MS-MS)
    
  • Waters Acquity QDa (UPLC-MS)
    
  • MD SpectraMax 384 (analyte detection)
    
  • Agilent HP1100/1200 (HPLC +/- MS Systems)
    
  • Waters Semi-Prep (HPLC)
    
  • Beckman Biomek NXP
    
  • Genevac EZ2-Plus evaporator
    

Sygnature supports a range of biomarker studies, with access to human and animal tissue, and is experience in the application of multiplex qPCR and multi-analyte detection technologies to monitor established biomarkers or to discover new biomarker candidates relevant to the project of interest. Using the Luminex assay platform, up to 50 analytes may be simultaneously measured per sample and custom analytes panels can be designed to suit client needs. Multiplexing offers increased throughput, facilitating more rapid assay development and conserving precious biological samples. Sygnature has a human tissue license and an in-house blood donor panel to allow easy access and storage of human blood and tissues, in addition to access to both healthy and diseased primary cells.

Sygnature's team has many years of knowledge and experience gained within major pharmaceutical companies to prosecute high-throughput screening (HTS) for hit identification purposes. HTS is project managed by Sygnature and undertaken through a strategic alliance partner. Sygnature develops and miniaturises appropriate HTS assays in 1,536-well format before transferring them to our CRO partner with >230K compound library (ex-major pharmaceutical company collection). We are able to undertake medium-throughput screening (MTS) at Sygnature using our proprietary fragment-based library of ~1,000 compounds, and a curated, structurally-diverse set of ~14,000 commercial compounds. The HTS and MTS data outputs are thereafter stored and analysed at Sygnature using the powerful Dotmatics Informatics systems.

Sygnature's Bioscience group has broad experience in handling a range of isolated biological targets across a diverse range of therapeutic areas from oncology to neuroscience. Assays can be developed and optimised to provide pivotal in vitro data for understanding structure-activity relationships (SAR) during medicinal chemistry-driven compound optimisation; as well as ex vivo assays to support pharmacodynamic model interpretation. The group is able to establish target-specific, or assay-specific counter-screens to validate early hit compounds, in particular to detect false positives and run detailed enzyme inhibition studies. Sygnature is able to utilise a diverse range of in-house assay detection technologies including colourimetric, fluorescence, luminescence and radiometric based assays, for which the company has the required radiochemical licensing.

In the absence of commercially-available reagents, Sygnature Discovery works with Peak Proteins, its sister company in Sygil Group, to provide construct design, protein expression and protein purification of the required protein to initiate projects based on novel biological targets and/or mechanisms. Proteins can be supplied in formats suitable for in vitro assays, high-throughput screening, biophysics assays, in vivo studies or X-ray crystallography, and may be expressed in E.coli, baculovirus/insect cells and mammalian cells, as appropriate. Peak Proteins works at scales from a few millilitres, to screen constructs, up to more than 20L to generate larger quantities of proteins. Sygnature works closely with Peak to ensure prompt delivery of the required protein or cell line for incorporation into an integrated drug discovery project.