Sussex Drug Discovery Centre (SDDC)

Assay development & compound screening

The group is able to provide medium/high throughput assays to screen focussed and fragment libraries and support ongoing hit to lead and lead optimisation programs. Assays can be ‘off the shelf’ or generated de novo including the provision of recombinant proteins or cellular reagents. The molecular pharmacology laboratories have proven expertise in:

 

Cell-based screening

The SDDC has a dedicated cell culture facility routinely culturing primary human and rodent cells (epithelial, neuronal) and a wide range of native and recombinant cell lines.

Plate readers for fluorescence (FlexStation, PHERAStar FS) and label free (Corning Epic) readouts enable medium/high throughput screening for ion channels, GPCRs and enzymes using a variety of formats including:

  • Calcium mobilisation
  • Membrane potential
  • Halide flux
  • Fluorescence polarisation (FP)
  • Time resolved fluorescence (TRF)
  • Fluorescence resonance energy transfer (FRET)
Cell free screening

The group has extensive experience in developing cell free binding and functional assays for all major target classes. This ranges from the generation of recombinant protein through to running compound and fragment screens in 96 and 384 well formats with fluorescence and radioactive endpoints.

Electrophysiology

Electrophysiology is a key strength of the SDDC enabling the effective prosecution of all types of ion channel targets. The group has multiple formats which can be applied to both primary and recombinant cell systems to screen and biophysically characterise novel ion channel modulators – these include:

  • Conventional whole-cell and single channel patch clamp rigs fitted with rapid and low volume perfusion systems (Dynaflow & RSC 200)
  • Medium throughput automated planar patch clamp system (Qpatch 16X)
  • Ussings chamber epithelial ion transport

DMPK

To help avoid costly failures during development it is essential that a pre-clinical candidate should exhibit not only high potency but also possess good safety and DMPK profiles. Feeding into the discovery programs from the earliest stages we routinely determine LogD and thermodynamic solubility, metabolic stability in microsomes and CYP450 inhibition across a number of species. To assist our Neurodegeneration programs our portfolio of In Vitro assays is soon to include an assessment of PgP liability in MDR-1 transfected MDCK cells and tissue/plasma binding in order to estimate bioavailability and free compound concentrations to support dosing regimens.