Perspectives on NMR in drug discovery: a technique comes of age

In the past decade, the potential of harnessing the ability of nuclear magnetic resonance (NMR) spectroscopy to monitor intermolecular interactions as a tool for drug discovery has been increasingly appreciated in academia and industry. In this Perspective, we highlight some of the major applications of NMR in drug discovery, focusing on hit and lead generation, and provide a critical analysis of its current and potential utility.

[1]  L. Kay,et al.  Solution NMR spectroscopy beyond 25 kDa. , 1997, Current opinion in structural biology.

[2]  Wolfgang Jahnke,et al.  Perspectives of biomolecular NMR in drug discovery: the blessing and curse of versatility , 2007, Journal of biomolecular NMR.

[3]  Brian K. Shoichet,et al.  Virtual screening of chemical libraries , 2004, Nature.

[4]  Ray Freeman,et al.  Fast reconstruction of four-dimensional NMR spectra from plane projections , 2004, Journal of biomolecular NMR.

[5]  D C Fry,et al.  Applications of biomolecular NMR to drug discovery. , 2000, Drug design and discovery.

[6]  O Hindsgaul,et al.  Saturation transfer difference NMR spectroscopy for identifying ligand epitopes and binding specificities. , 2004, Ernst Schering Research Foundation workshop.

[7]  Ernest Giralt,et al.  NMR-based methods and strategies for drug discovery. , 2003, Chemical Society reviews.

[8]  Andrea Giachetti,et al.  Combining in silico tools and NMR data to validate protein-ligand structural models: application to matrix metalloproteinases. , 2005, Journal of medicinal chemistry.

[9]  Ernest Giralt,et al.  Disruption of the HIV-1 protease dimer with interface peptides: structural studies using NMR spectroscopy combined with [2-(13)C]-Trp selective labeling. , 2007, Biopolymers.

[10]  L. Kay,et al.  NMR studies of protein structure and dynamics. , 2005, Journal of magnetic resonance.

[11]  Stephen W. Fesik,et al.  One-Dimensional Relaxation- and Diffusion-Edited NMR Methods for Screening Compounds That Bind to Macromolecules , 1997 .

[12]  Nicola Mongelli,et al.  A general NMR method for rapid, efficient, and reliable biochemical screening. , 2003, Journal of the American Chemical Society.

[13]  L. Kay,et al.  NMR methods for the study of protein structure and dynamics. , 1997, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[14]  M. Sundström,et al.  Identification of compounds with binding affinity to proteins via magnetization transfer from bulk water* , 2000, Journal of biomolecular NMR.

[15]  I. Kuntz,et al.  The maximal affinity of ligands. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[16]  Marina Veronesi,et al.  Fluorine-NMR competition binding experiments for high-throughput screening of large compound mixtures. , 2002, Combinatorial chemistry & high throughput screening.

[17]  F. Lombardo,et al.  Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. , 2001, Advanced drug delivery reviews.

[18]  S. Korsmeyer,et al.  An inhibitor of Bcl-2 family proteins induces regression of solid tumours , 2005, Nature.

[19]  Ray Freeman,et al.  Reconstruction of the three-dimensional NMR spectrum of a protein from a set of plane projections , 2003, Journal of biomolecular NMR.

[20]  Volker Dötsch,et al.  In‐Cell NMR Spectroscopy , 2005, Methods in enzymology.

[21]  N J Sucher,et al.  Traditional Chinese medicines with caspase-inhibitory activity. , 2006, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[22]  Lance M. Westerhoff,et al.  A critical assessment of the performance of protein-ligand scoring functions based on NMR chemical shift perturbations. , 2007, Journal of medicinal chemistry.

[23]  A Medek,et al.  An approach for high-throughput structure determination of proteins by NMR spectroscopy , 2000, Journal of biomolecular NMR.

[24]  Zartler Er,et al.  Protein NMR-based screening in drug discovery. , 2006 .

[25]  K. Pervushin,et al.  Impact of Transverse Relaxation Optimized Spectroscopy (TROSY) on NMR as a technique in structural biology , 2000, Quarterly Reviews of Biophysics.

[26]  Maurizio Pellecchia,et al.  A fragment-based approach for the discovery of isoform-specific p38alpha inhibitors. , 2007, ACS chemical biology.

[27]  Wim G. J. Hol,et al.  Structural genomics for science and society , 2000, Nature Structural Biology.

[28]  Hartmut Oschkinat,et al.  Determination of solid-state NMR structures of proteins by means of three-dimensional 15N-13C-13C dipolar correlation spectroscopy and chemical shift analysis. , 2003, Biochemistry.

[29]  Joanna Owens,et al.  Chris Lipinski discusses life and chemistry after the Rule of Five. , 2003 .

[30]  S. Homans,et al.  Water, water everywhere--except where it matters? , 2007, Drug discovery today.

[31]  Nicola Mongelli,et al.  NMR-based quality control approach for the identification of false positives and false negatives in high throughput screening. , 2006, Current drug discovery technologies.

[32]  M. Baldus,et al.  High-resolution solid-state NMR applied to polypeptides and membrane proteins. , 2003, Accounts of chemical research.

[33]  Richard M. Myers,et al.  Recent progress and perspectives for GLAO and MOAO , 2010, Astronomical Telescopes + Instrumentation.

[34]  M. Schubert,et al.  Structure of a protein determined by solid-state magic-angle-spinning NMR spectroscopy , 2002, Nature.

[35]  Christopher M. Dobson,et al.  Molecular recycling within amyloid fibrils , 2005, Nature.

[36]  Ernest Giralt,et al.  Identification by 19F NMR of Traditional Chinese Medicinal Plants Possessing Prolyl Oligopeptidase Inhibitory Activity , 2006, Chembiochem : a European journal of chemical biology.

[37]  Dawoon Jung,et al.  Discovery of a novel class of reversible non-peptide caspase inhibitors via a structure-based approach. , 2005, Journal of medicinal chemistry.

[38]  Valler,et al.  Diversity screening versus focussed screening in drug discovery. , 2000, Drug discovery today.

[39]  Ernest Giralt,et al.  A fast and robust 19F NMR-based method for finding new HIV-1 protease inhibitors. , 2006, Bioorganic & medicinal chemistry letters.

[40]  Stephen R. Johnson,et al.  Molecular properties that influence the oral bioavailability of drug candidates. , 2002, Journal of medicinal chemistry.

[41]  Harald Schwalbe,et al.  NMR Backbone Assignment of a Protein Kinase Catalytic Domain by a Combination of Several Approaches: Application to the Catalytic Subunit of cAMP‐Dependent Protein Kinase , 2004, Chembiochem : a European journal of chemical biology.

[42]  Mei Hong,et al.  Oligomeric structure, dynamics, and orientation of membrane proteins from solid-state NMR. , 2006, Structure.

[43]  P. Hajduk,et al.  NMR-based screening in drug discovery , 1999, Quarterly Reviews of Biophysics.

[44]  P. Hajduk Fragment-based drug design: how big is too big? , 2006, Journal of medicinal chemistry.

[45]  Gerhard Wagner,et al.  Quantitative NMR analysis of the protein G B1 domain in Xenopus laevis egg extracts and intact oocytes , 2006, Proceedings of the National Academy of Sciences.

[46]  Horst Kessler,et al.  NMR-based screening technologies. , 2003, Drug discovery today.

[47]  Claudio Dalvit,et al.  Multi-selective one dimensional proton NMR experiments for rapid screening and binding affinity measurements. , 2003, Combinatorial chemistry & high throughput screening.

[48]  Wolfgang Jahnke,et al.  Fragment-based Approaches in Drug Discovery: JAHNKE: FRAGMENT-BASED APPROACHES IN DRUG DISCOVERY O-BK , 2006 .

[49]  Harald Schwalbe,et al.  NMR characterization of kinase p38 dynamics in free and ligand-bound forms. , 2006, Angewandte Chemie.

[50]  T. O'Brien,et al.  Fragment-based drug discovery. , 2004, Journal of medicinal chemistry.

[51]  M. Uesugi,et al.  [Discovering high-affinity ligands for proteins: SAR by NMR]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[52]  Christopher W Murray,et al.  Fragment-based lead discovery: leads by design. , 2005, Drug discovery today.

[53]  R. Freeman,et al.  Projection-reconstruction of three-dimensional NMR spectra. , 2003, Journal of the American Chemical Society.

[54]  R. Harrington Part II , 2004 .

[55]  R Kiplin Guy,et al.  Synthesis and testing of a focused phenothiazine library for binding to HIV-1 TAR RNA. , 2006, Chemistry & biology.

[56]  A. Hopkins,et al.  Ligand efficiency: a useful metric for lead selection. , 2004, Drug discovery today.

[57]  Maurizio Pellecchia,et al.  SAR by ILOEs: an NMR-based approach to reverse chemical genetics. , 2006, Chemistry.

[58]  Alexander Shekhtman,et al.  Semisynthesis of a segmental isotopically labeled protein splicing precursor: NMR evidence for an unusual peptide bond at the N-extein-intein junction. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[59]  Maurizio Pellecchia,et al.  A Structure-based Approach to Retinoid X Receptor-α Inhibition* , 2006, Journal of Biological Chemistry.

[60]  I. Kuntz,et al.  Structure-based discovery of inhibitors of thymidylate synthase. , 1993, Science.

[61]  Marina Veronesi,et al.  Competition binding experiments for rapidly ranking lead molecules for their binding affinity to human serum albumin. , 2002, Combinatorial chemistry & high throughput screening.

[62]  Horst Kessler,et al.  Diffusion NMR Spectroscopy: Folding and Aggregation of Domains in p53 , 2005, Chembiochem : a European journal of chemical biology.

[63]  P. Hajduk,et al.  SOS-NMR: a saturation transfer NMR-based method for determining the structures of protein-ligand complexes. , 2004, Journal of the American Chemical Society.

[64]  D. J. Triggle,et al.  Comprehensive medicinal chemistry II , 2006 .

[65]  P. Hajduk,et al.  NMR in Pharmacokinetic and Pharmacodynamic Profiling , 2005, Chembiochem : a European journal of chemical biology.

[66]  Toshio Yamazaki,et al.  Segmental Isotope Labeling for Protein NMR Using Peptide Splicing , 1998 .

[67]  Kurt Wüthrich,et al.  NMR solution structure determination of membrane proteins reconstituted in detergent micelles , 2003, FEBS letters.

[68]  Daniel A Erlanson,et al.  Tethering: fragment-based drug discovery. , 2004, Annual review of biophysics and biomolecular structure.

[69]  Edward Olejniczak,et al.  Characterization of protein-ligand interactions by high-resolution solid-state NMR spectroscopy. , 2004, Journal of the American Chemical Society.

[70]  Dawoon Jung,et al.  NMR-based techniques in the hit identification and optimisation processes , 2004, Expert opinion on therapeutic targets.

[71]  P. Hajduk,et al.  Druggability indices for protein targets derived from NMR-based screening data. , 2005, Journal of medicinal chemistry.

[72]  Christopher W Murray,et al.  Fragment-based lead discovery using X-ray crystallography. , 2005, Journal of medicinal chemistry.

[73]  B. Meyer,et al.  Group epitope mapping by saturation transfer difference NMR to identify segments of a ligand in direct contact with a protein receptor. , 2001, Journal of the American Chemical Society.

[74]  Renaldo Mendoza,et al.  ALARM NMR: a rapid and robust experimental method to detect reactive false positives in biochemical screens. , 2005, Journal of the American Chemical Society.

[75]  Markus Schade,et al.  NMR fragment screening: tackling protein-protein interaction targets. , 2005, Current opinion in drug discovery & development.

[76]  C. Lipinski Drug-like properties and the causes of poor solubility and poor permeability. , 2000, Journal of pharmacological and toxicological methods.

[77]  G. Fogliatto,et al.  WaterLOGSY as a method for primary NMR screening: Practical aspects and range of applicability , 2001, Journal of biomolecular NMR.

[78]  Nicola Mongelli,et al.  Polyfluorinated amino acids for sensitive 19F NMR-based screening and kinetic measurements. , 2007, Journal of the American Chemical Society.

[79]  A. Bax Weak alignment offers new NMR opportunities to study protein structure and dynamics , 2003, Protein science : a publication of the Protein Society.

[80]  Edward R Zartler,et al.  The effect of relaxation on the epitope mapping by saturation transfer difference NMR. , 2003, Journal of magnetic resonance.

[81]  Horst Kessler,et al.  Orientational properties of stretched polystyrene gels in organic solvents and the suppression of their residual 1H NMR signals. , 2005, Journal of the American Chemical Society.

[82]  Harald Schwalbe,et al.  Combined solid state and solution NMR studies of alpha,epsilon-15N labeled bovine rhodopsin. , 2007, Journal of biomolecular NMR.

[83]  Andrea Giachetti,et al.  Exploring the subtleties of drug-receptor interactions: the case of matrix metalloproteinases. , 2007, Journal of the American Chemical Society.

[84]  Kurt Wüthrich,et al.  Nmr in drug discovery , 2002, Nature Reviews Drug Discovery.

[85]  Maurizio Pellecchia,et al.  Targeting apoptosis via chemical design: inhibition of bid-induced cell death by small organic molecules. , 2004, Chemistry & biology.

[86]  Karl A. Walter,et al.  ChemInform Abstract: Discovery of Potent Nonpeptide Inhibitors of Stromelysin Using SAR by NMR. , 1997 .

[87]  Maurizio Pellecchia,et al.  NMR-based structural characterization of large protein-ligand interactions , 2002, Journal of biomolecular NMR.

[88]  Volker Dötsch,et al.  Combination of cell‐free expression and NMR spectroscopy as a new approach for structural investigation of membrane proteins , 2006, Magnetic resonance in chemistry : MRC.

[89]  J. T. Metz,et al.  Ligand efficiency indices as guideposts for drug discovery. , 2005, Drug discovery today.

[90]  William S. Price,et al.  Pulsed-field gradient nuclear magnetic resonance as a tool for studying translational diffusion: part II. Experimental aspects , 1998 .

[91]  P. Hajduk,et al.  A decade of fragment-based drug design: strategic advances and lessons learned , 2007, Nature Reviews Drug Discovery.

[92]  T. James,et al.  NMR-based characterization of phenothiazines as a RNA binding scaffold. , 2004, Journal of the American Chemical Society.

[93]  Claudio Dalvit,et al.  Ligand- and substrate-based 19F NMR screening: Principles and applications to drug discovery , 2007 .

[94]  W. Moore Maximizing discovery efficiency with a computationally driven fragment approach. , 2005, Current opinion in drug discovery & development.

[95]  R. Stroud,et al.  Site-directed ligand discovery. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[96]  I. Bertini,et al.  Metabolomics: available results, current research projects in breast cancer, and future applications. , 2007, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[97]  M. McCoy,et al.  Spatial localization of ligand binding sites from electron current density surfaces calculated from NMR chemical shift perturbations. , 2002, Journal of the American Chemical Society.

[98]  Gang Liu,et al.  Discovery of a potent, selective protein tyrosine phosphatase 1B inhibitor using a linked-fragment strategy. , 2003, Journal of the American Chemical Society.

[99]  Horst Kessler,et al.  Structure Refinement of Cyclosporin A in Chloroform by Using RDCs Measured in a Stretched PDMS‐Gel , 2005, Chembiochem : a European journal of chemical biology.

[100]  P. Hajduk,et al.  Integration of NMR and high-throughput screening. , 2002, Combinatorial chemistry & high throughput screening.

[101]  R M Stroud,et al.  A new class of HIV-1 protease inhibitor: the crystallographic structure, inhibition and chemical synthesis of an aminimide peptide isostere. , 1996, Bioorganic & medicinal chemistry.

[102]  Marc Baldus,et al.  Investigation of Ligand‐Receptor Systems by High‐Resolution Solid‐State NMR: Recent Progress and Perspectives , 2005, Archiv der Pharmazie.

[103]  L. Kay,et al.  A robust and cost-effective method for the production of Val, Leu, Ile (δ1) methyl-protonated 15N-, 13C-, 2H-labeled proteins , 1999, Journal of biomolecular NMR.

[104]  Ajay,et al.  The SHAPES strategy: an NMR-based approach for lead generation in drug discovery. , 1999, Chemistry & biology.

[105]  M. Congreve,et al.  Fragment-based lead discovery , 2004, Nature Reviews Drug Discovery.

[106]  L. Kay,et al.  The use of 2H, 13C, 15N multidimensional NMR to study the structure and dynamics of proteins. , 1998, Annual review of biophysics and biomolecular structure.

[107]  W. Jahnke,et al.  Spin label enhanced NMR screening. , 2001, Journal of the American Chemical Society.

[108]  Horst Kessler,et al.  Phosphorus NMR spectroscopy as a versatile tool for compound library screening. , 2008, Angewandte Chemie.

[109]  Ernest Giralt,et al.  A tetraguanidinium ligand binds to the surface of the tetramerization domain of protein P53. , 2004, Angewandte Chemie.

[110]  Wyss Df,et al.  NMR-based approaches for lead discovery. , 2002 .

[111]  W. Jahnke,et al.  Protein NMR in biomedical research , 2004, Cellular and Molecular Life Sciences CMLS.

[112]  Edward R. Zartler,et al.  Fragment-Based Drug Discovery , 2008 .

[113]  S. Züger,et al.  Intein-based biosynthetic incorporation of unlabeled protein tags into isotopically labeled proteins for NMR studies , 2005, Nature Biotechnology.

[114]  Kristiina Vuori,et al.  Development of molecular probes for second-site screening and design of protein tyrosine phosphatase inhibitors. , 2007, Journal of medicinal chemistry.

[115]  Maurizio Pellecchia,et al.  The Nuclear Overhauser Effect in the lead identification process. , 2006, Current drug discovery technologies.

[116]  P. Hajduk,et al.  Utilization of NMR-derived fragment leads in drug design. , 2005, Methods in enzymology.

[117]  J. Prestegard,et al.  Residual dipolar couplings in structure determination of biomolecules. , 2004, Chemical reviews.

[118]  K Wüthrich Nuclear magnetic resonance studies of internal mobility in globular proteins. , 1981, Biochemical Society symposium.

[119]  Dawoon Jung,et al.  Efficient synthetic inhibitors of anthrax lethal factor. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[120]  Wolfgang Jahnke,et al.  Second-Site NMR Screening with a Spin-Labeled First Ligand , 2000 .

[121]  Claudio Dalvit,et al.  Fluorine-NMR experiments for high-throughput screening: theoretical aspects, practical considerations, and range of applicability. , 2003, Journal of the American Chemical Society.

[122]  P. Hajduk,et al.  Privileged molecules for protein binding identified from NMR-based screening. , 2000, Journal of medicinal chemistry.

[123]  Agnieszka Bronowska,et al.  Contribution of ligand desolvation to binding thermodynamics in a ligand-protein interaction. , 2006, Angewandte Chemie.

[124]  William S. Price,et al.  Pulsed-Field Gradient Nuclear Magnetic Resonance as a Tool for Studying Translational Diffusion. Part 1. Basic Theory , 1997 .

[125]  Daniel A Erlanson,et al.  Making drugs on proteins: site-directed ligand discovery for fragment-based lead assembly. , 2004, Current opinion in chemical biology.

[126]  T. James,et al.  Detecting ligand binding to a small RNA target via saturation transfer difference NMR experiments in D(2)O and H(2)O. , 2002, Journal of the American Chemical Society.

[127]  I D Kuntz,et al.  Structure of a non-peptide inhibitor complexed with HIV-1 protease. Developing a cycle of structure-based drug design. , 1997, The Journal of biological chemistry.

[128]  W. Pitt,et al.  Identification of novel fragment compounds targeted against the pY pocket of v‐Src SH2 by computational and NMR screening and thermodynamic evaluation , 2007, Proteins.

[129]  Nicola Mongelli,et al.  Reliable high-throughput functional screening with 3-FABS. , 2004, Drug discovery today.

[130]  Gregg Siegal,et al.  TINS, target immobilized NMR screening: an efficient and sensitive method for ligand discovery. , 2005, Chemistry & biology.

[131]  H. Jhoti,et al.  The Discovery of Novel Protein Kinase Inhibitors by Using Fragment‐Based High‐Throughput X‐ray Crystallography , 2005, Chembiochem : a European journal of chemical biology.

[132]  Ray Freeman,et al.  Fast multi-dimensional Hadamard spectroscopy. , 2003, Journal of magnetic resonance.

[133]  D. Kern,et al.  The role of dynamics in allosteric regulation. , 2003, Current opinion in structural biology.

[134]  Harald Schwalbe,et al.  Biomolecular NMR: a chaperone to drug discovery , 2006, Current Opinion in Chemical Biology.

[135]  Ernest Giralt,et al.  The Natural Product Berberine is a Human Prolyl Oligopeptidase Inhibitor , 2007, ChemMedChem.

[136]  Jeffrey W. Peng,et al.  Applications of SHAPES screening in drug discovery. , 2002, Combinatorial chemistry & high throughput screening.

[137]  Nicola Mongelli,et al.  Sensitivity improvement in 19F NMR-based screening experiments: theoretical considerations and experimental applications. , 2005, Journal of the American Chemical Society.

[138]  Thomas Szyperski,et al.  Reduced-dimensionality NMR spectroscopy for high-throughput protein resonance assignment , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[139]  Irwin D. Kuntz,et al.  Development and validation of a modular, extensible docking program: DOCK 5 , 2006, J. Comput. Aided Mol. Des..

[140]  Maurizio Pellecchia,et al.  Structure–activity relationships by interligand NOE-based design and synthesis of antiapoptotic compounds targeting Bid , 2006, Proceedings of the National Academy of Sciences.

[141]  Edward R Zartler,et al.  Fragonomics: fragment-based drug discovery. , 2005, Current opinion in chemical biology.

[142]  R. Riek,et al.  Attenuated T2 relaxation by mutual cancellation of dipole-dipole coupling and chemical shift anisotropy indicates an avenue to NMR structures of very large biological macromolecules in solution. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[143]  P. Bartlett,et al.  Exploiting chemical diversity for drug discovery , 2006 .

[144]  Horst Kessler,et al.  NMR-based screening: a powerful tool in fragment-based drug discovery. , 2007 .

[145]  Bernd Meyer,et al.  Characterization of Ligand Binding by Saturation Transfer Difference NMR Spectroscopy. , 1999, Angewandte Chemie.

[146]  H. Schwalbe,et al.  Cross-correlated relaxation for measurement of angles between tensorial interactions. , 2001, Methods in enzymology.

[147]  C. Sanders,et al.  Solution NMR of membrane proteins: practice and challenges , 2006, Magnetic resonance in chemistry : MRC.

[148]  Harald Schwalbe,et al.  How Much NMR Data Is Required To Determine a Protein–Ligand Complex Structure? , 2005, Chembiochem : a European journal of chemical biology.

[149]  Harald Schwalbe,et al.  Combined solid state and solution NMR studies of α,ɛ-15N labeled bovine rhodopsin , 2007 .

[150]  Horst Kessler,et al.  Conformational Analysis of Drugs by Nuclear Magnetic Resonance Spectroscopy , 2007 .

[151]  Maurizio Pellecchia,et al.  A fragment-based approach for the discovery of isoform-specific p38? inhibitors , 2007 .

[152]  Ivano Bertini,et al.  Conformational variability of matrix metalloproteinases: beyond a single 3D structure. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[153]  Wolfgang Jahnke Faculty Opinions recommendation of Fluorine-NMR experiments for high-throughput screening: theoretical aspects, practical considerations, and range of applicability. , 2003 .

[154]  P. Hajduk,et al.  Discovery of a potent inhibitor of the antiapoptotic protein Bcl-xL from NMR and parallel synthesis. , 2006, Journal of medicinal chemistry.

[155]  Jens Meiler,et al.  The INPHARMA method: protein-mediated interligand NOEs for pharmacophore mapping. , 2005, Angewandte Chemie.

[156]  Hugo O Villar,et al.  Using NMR for ligand discovery and optimization. , 2004, Current opinion in chemical biology.

[157]  P. Hajduk,et al.  High-throughput nuclear magnetic resonance-based screening. , 1999, Journal of medicinal chemistry.

[158]  M. Congreve,et al.  A 'rule of three' for fragment-based lead discovery? , 2003, Drug discovery today.

[159]  Gaohua Liu,et al.  NMR data collection and analysis protocol for high-throughput protein structure determination. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[160]  D Cowburn,et al.  Segmental isotopic labeling using expressed protein ligation. , 2001, Methods in enzymology.

[161]  Ivano Bertini,et al.  Experimentally exploring the conformational space sampled by domain reorientation in calmodulin. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[162]  Ray Freeman,et al.  Projection-reconstruction technique for speeding up multidimensional NMR spectroscopy. , 2004, Journal of the American Chemical Society.

[163]  Karen N. Allen,et al.  Design, synthesis, and characterization of a potent xylose isomerase inhibitor, D-threonohydroxamic acid, and high-resolution X-ray crystallographic structure of the enzyme-inhibitor complex. , 1995, Biochemistry.

[164]  Jeffrey W. Peng,et al.  Application of NMR SHAPES screening to an RNA target. , 2003, Journal of the American Chemical Society.

[165]  Koh Takeuchi,et al.  NMR studies of protein interactions. , 2006, Current opinion in structural biology.

[166]  Claudio Dalvit,et al.  NMR screening techniques in drug discovery and drug design , 2002 .