Lead Discovery and Optimization

Complete workflow from fragment library quality check, cocktail design and automated hit identification to quantitative assessment of binding (Kd) and the 3D analysis of ligands.

Library Definition

• Quality check of the fragment library with (Mnova Gears,Verify&qNMR)
• 鸡尾酒设计MixDesign

Library Screening

• Handling of screening sample withSampleJet
  
• Standard and customizable¹H and¹⁹F NMR screening experiments can be run in automation
  
• High¹H and¹⁹F sensitivity with CryoProbes (QCIF)
  
• Automated¹H and¹⁹F hit detection with the integration of FBS with Mnova Screen

Structural Biology

Mnova Bindingautomatically processes protein-ligand titration 2D HSQC, tracks the peak movement and calculates the Kd’s for multiple peaks.

Lead Optimization

The 3D conformations adopted by a free ligand in solution affects binding affinity for the pharmaceutical target. Their structural analysis helps to speed up lead optimization.
Mnova StereoFittergenerates 3D conformational populations using experimental NMR constraints (qNOE, chemical shift and J coupling).

• Automatic acquisition of¹H and¹⁹F NMR experiments with IconNMR. Default experiments can be found in the Bruker library:
  -¹H NMR screening experiments with buffer/water suppression:¹H 1D, Saturation Transfer Difference,
  waterLOGSY,
  - T2 and T1rho
  -¹⁹F screening experiment with decoupling.
  
• TopSpin Fragment Based Screening functionality bridges acquisition with analysis. Upon acquisition data are automatically analysed byMnova Screen. The results are reported in Mnova screen in a Power Point like format.
  
• Protein-observed screening uses the Chemical Shifts Perturbation (CSP) method. H-C correlation experiment (HSQC) of the target with increasing amounts of ligand are performed. The data is analysed usingMnova Binding, which allow visualisation of the data and the determination of Kd.
  
• Mnova StereoFittercomputes the probability of 3D structural configurations and/or conformations, based on various forms of NMR experimental data input. Currently, StereoFitter can accept four distinct types of input in order to calculate the best 3D structure candidate(s): NOEs, RDCs, Js, and chemical shifts.
  
• The workflows can be enriched with aMnova DB, to store, for example, the screening data and 1D spectra of fragments which are frequently used during the fragment screening experiments.
  
  

• NMR sensitivity to fragment binding is ideally matching the range of weak affinities where fragments are expected (Kd, the dissociation constant, is in the range of micromolar [μM] to a few millimolar [mM])
• The NMR screening experiments offers both binding and ligands/s structural information
• NMR is unique in determining experimentally the ligand’s conformation in solution (3D analysis), unraveling the conformational change between solution and bound state
• The selectivity and wide dispersion intrinsic to¹⁹F NMR,使¹⁹F screening an attractive method
  - Up to 30 fragments in a mixture
  - High sensitivity because of strong T2 effects
  - Selectivity: no fluor in protein samples
  -¹⁹F fragment libraries commercially available.
• As a typical screening can generate hundreds of spectra. Automated and robust data analysis software enable lead discovery and optimisation with minimal manual operations and superior reproducibility.
• All needed software tools are now included in the new Lead discovery and Optimisation Suite. Contact us for more information!