Theory and Applications of NMR-Based Screening in Pharmaceutical Research

Lepre, Christopher A.; Moore, Jonathan M.; Peng, Jeffrey W.

doi:10.1021/cr030409h

Cited by 298 publications

(194 citation statements)

References 129 publications

Supporting

Mentioning

190

Contrasting

Unclassified

Order By: Relevance

“…The relative STD effect was calculated for each signal as the difference between the intensity (expressed as S/N ratio) of one signal in the on-resonance STD spectrum and that of the same signal in the off-resonance NMR spectrum divided by the intensity of the same signal in the offresonance spectrum. BMI values were obtained by using Equation (1).…”

Section: Methodsmentioning

confidence: 99%

“…[1] In particular, saturation transfer difference (STD) spectroscopy [2] offers several advantages: a modest requirement of an unlabeled biological target, virtually without upper limits on its size; amenability to screening mixtures of putative ligands in the presence of non-ligands, molecules that target viruses, [3] and membrane-bound receptors on living cells; [4] and rapid access to relevant information on ligand epitope maps. [5] A main limitation, common to all ligand-based approaches for NMR screening, is that no direct information can be gathered on the intervening receptor surface, and therefore on the mode and site of binding, even if an indirect method (SOS-NMR; SOS: structural information using Overhauser effects and selective labeling) based on selective protein labeling has been devised.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Differential‐Frequency Saturation Transfer Difference NMR Spectroscopy Allows the Detection of Different Ligand–DNA Binding Modes

et al. 2005

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Differential‐Frequency Saturation Transfer Difference NMR Spectroscopy Allows the Detection of Different Ligand–DNA Binding Modes

et al. 2005

View full text Add to dashboard Cite

show abstract

“…Pulse sequences of this type have the advantage that they do not cause sample heating, attenuate protein signals, cause metabolite protein-mediated saturations or suppress signals from exchangeable protons, all of which are common problems with otherwise robust saturation-based approaches. These pulse sequences are very popular and have been used in fields such as food analysis, 4 metabo(l/n)omics, 5,6 drug discovery, 7 and environmental studies. 8 Within this class, sequences that use the excitation sculpting principle 9 are particularly efficient.…”

Section: 1mentioning

confidence: 99%

Robust NMR water signal suppression for demanding analytical applications

Aguilar

Kenwright

2016

Analyst

View full text Add to dashboard Cite

Publisher's copyright statement:Additional information: Use policyThe full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-pro t purposes provided that:• a full bibliographic reference is made to the original source • a link is made to the metadata record in DRO • the full-text is not changed in any way The full-text must not be sold in any format or medium without the formal permission of the copyright holders.Please consult the full DRO policy for further details. We describe the design and application of robust, general-purpose water signal suppression pulse sequences well suited to chemometric work. Such pulse sequences need to deal well with pulse mis-calibrations, radiation damping, chemical exchange, and the presence of sample inhomogeneities, as well as with significant variations in sample characteristics such as pH, ionic strength, relaxation characteristics and molecular weight. Of course, such pulse sequences should produce undistorted lineshapes and baselines and work well both under automation and in the hands of non-experts. As an example, one such pulse sequences, Robust-5, will be presented. This new pulse sequence meets those criteria and is able to reduce a 50 M proteo water signal down to a 0.9 mM level, without fine tuning, and under automation, and it is therefore well suited to the most demanding of analytical applications.

show abstract

“…One of the major disadvantages of this technique is that the protein must be stable over the long data acquisition process. [37] Wang et al screened a 10,000-member custom made fragment library against 15 N-labelled BACE-1, a target currently thought to be important for the treatment of Alzheimers disease, using HSQC NMR. [38] They identified a thiourea fragment with a K d of 550 µM, which was rapidly optimised using a combination of HSQC NMR and a functional assay to improve affinity by a factor of 36, to give a K d of 15 µM.…”

Section: Challenges For Fbdd: Fragment Screeningmentioning

confidence: 99%

Kinetic Template‐Guided Tethering of Fragments

2012

View full text Add to dashboard Cite

This thesis is composed of two separate projects: Kinetic Template-Guided Tethering of Fragments and Design and Synthesis of a Chemical Probe to Dissect the Cellular Signalling Cascade leading to Cyclin D1 Degradation after DNA Damage. Kinetic Template-Guided Tethering of FragmentsThe development of a novel methodology for the site-directed discovery of small molecule, protein-binding ligands is described. The protein of interest, with a cysteine thiol (either native or engineered) adjacent to the desired binding pocket, is incubated with mixtures of low molecular weight compounds (fragments) modified with either an acrylamide or a vinyl sulfonamide capture group. Any ligand within the mixture that binds within the pocket brings the capture group into close proximity with the cysteine thiol, promoting a conjugate addition reaction at an increased rate over the background reaction. The capture reaction is designed to be slow, such that during the time course of an experiment, no adduct formation is observed unless the reaction is templated by the protein. By this method, binding ligands are rapidly identified by mass spectrometry analysis of the crude reaction mixtures. The methodology has been termed 'kinetic template-guided tethering'. Design and Synthesis of a Chemical Probe to Dissect the Cellular Signalling Cascade leading to Cyclin D1Degradation after DNA Damage An inhibitor described within the literature was found to attenuate the reduction of cyclin D1 after DNA damage to cells. In order to implement a two-step chemical proteomics strategy to find the molecular target of this inhibitor, a synthesis of the compound with an alkyne appendage was required. The alkyne acts as a functional handle for attachment of a reporter molecule in cells via the Huisgen cycloaddition ('Click') reaction. The design and synthesis of this inhibitor with the alkyne appendage is described.

show abstract

Theory and Applications of NMR-Based Screening in Pharmaceutical Research

Cited by 298 publications

References 129 publications

Differential‐Frequency Saturation Transfer Difference NMR Spectroscopy Allows the Detection of Different Ligand–DNA Binding Modes

Differential‐Frequency Saturation Transfer Difference NMR Spectroscopy Allows the Detection of Different Ligand–DNA Binding Modes

Robust NMR water signal suppression for demanding analytical applications

Kinetic Template‐Guided Tethering of Fragments

Contact Info

Product

Resources

About