N -Acyl phenylalanine analogues as potent small molecule VLA-4 antagonists

Chen, Li; Tilley, Jefferson; Huang, Tai-Nan; Miklowski, Dorota; Trilles, Richard; Guthrie, Robert; Luk, Kin-chun; Hanglow, A. C.; Rowan, Karen; Schwinge, Virginia; Wolitzky, Barry A.

doi:10.1016/s0960-894x(00)00088-3

Cited by 21 publications

(7 citation statements)

References 4 publications

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“…Small-molecule blockers of protein-protein interactions have been successfully identifi ed using a variety of screening techniques. Sandwich ELISAs have been applied widely, for example for the measurement of the p53-mdm2, 14 the VLA4-VCAM, 15 and the B7.1-CD28 16 protein-protein interaction. Fluorescence polarization has also been used to measure the binding of a BH3 peptide to Bcl-XL 17 and a p53 peptide to mdm2.…”

Section: Protein Labeling and Storage Of Tracersmentioning

confidence: 99%

A Cy5-Labeled S100A10 Tracer Used to Identify Inhibitors of the Protein Interaction With Annexin A2

Reddy

Fischer

et al. 2010

ASSAY and Drug Development Technologies

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Protein-protein interactions are increasingly of interest as targets in small-molecule drug discovery. The interaction between the Ca2+- and phospholipid-binding protein Annexin A2 and its binding partner S100A10 has been implicated in angiogenesis and cancer metastasis. Here, we present a methodology to screen for inhibitors of this protein interaction. We developed a Cy5-labeled S100A10 tracer and showed by circular dichroism spectroscopy that the secondary structure is indistinguishable from that of non-labeled S100A10. This tracer was used to develop a binding assay based upon fluorescence resonance energy transfer to a Cy3-labeled Annexin A2 peptide ligand. The binding parameters matched those for unlabeled components as observed by equilibrium dialysis, which we determined separately, as well as those determined by isothermal titration calorimetry. Binding of labeled and unlabeled peptide was specific and mutually competitive. We used this assay for screening a small compound library derived by computational interrogation of the S100A10-binding pocket. Hits were obtained with IC(50) values in range of the IC(50) of the cognate Annexin A2 peptide ligand. Hits were subjected to an exact parallel assay measuring an unrelated protein-protein interaction (antigen-antibody). In this way, we identified genuine hits that inhibited the interaction between S100A10 and Annexin A2 but do not affect the fluorescence readout. These compounds are potentially of interest as candidates for further analysis and medical chemistry exploration. The simple assay format described here can be employed in early-stage exploration of other protein-protein interaction targets.

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Section: Protein Labeling and Storage Of Tracersmentioning

confidence: 99%

A Cy5-Labeled S100A10 Tracer Used to Identify Inhibitors of the Protein Interaction With Annexin A2

Reddy

Fischer

et al. 2010

ASSAY and Drug Development Technologies

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“…Within fibronectin, a leucine-aspartic acid-valine (LDV) triad was identified as the minimal essential sequence for binding [5], and within VCAM-1, a glutamine-isoleucine-aspartic acidserine (QIDS) sequence is required [6]. Peptides containing these sequences have been shown to inhibit α4β1 mediated *Address correspondence to this author at Wyeth Research, CN8000, Princeton, NJ 08543, USA; Tel: (732)-274-4025; Fax: (732)-274-4129; Email: hurynd@wyeth.com adhesion, and a number of groups have used these sequences [7][8][9][10][11] and others [12,13] as starting points for the design of small molecule antagonists [14][15][16].…”

Section: Identification Of a Novel Binding Epitopementioning

confidence: 99%

The Identification and Optimization of Orally Efficacious, Small Molecule VLA-4 Antagonists

Huryn¹,

Konradi²,

Ashwell³

et al. 2004

CTMC

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The identification of orally active, small molecule antagonists of the alpha4beta1 integrin, VLA-4, could lead to therapeutic agents with utility in a number of clinical settings, including asthma, multiple sclerosis and IBD. Starting from CDR3 sequences conserved among neutralizing alpha4 antibodies, peptides were identified that antagonized VLA-4 mediated adhesion in vitro. Through a series of structural modifications, these peptides evolved into small molecules that exhibited high potency and selectivity for VLA-4 in cell adhesion assays. Finally, through the optimization of physical and pharmacokinetic properties, compounds were identified that exhibited oral activity in animal models of asthma and multiple sclerosis.

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“…These compounds show tumour growth inhibition in tumour models and have progressed to early clinical-stage research 3. Finally, various investigational small-molecule blockers have been reported for the protein interaction between VLA4–VCAM,4 B7.1–CD28,5 oestrogen-related receptor-α and its co-activator,6 and protein kinase C-iota and its effector PAR6 7…”

Section: Introductionmentioning

confidence: 99%

Three‐Dimensional Pharmacophore Design and Biochemical Screening Identifies Substituted 1,2,4‐Triazoles as Inhibitors of the Annexin A2–S100A10 Protein Interaction

Reddy

Fischer

et al. 2012

ChemMedChem

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Protein interactions are increasingly appreciated as targets in small-molecule drug discovery. The interaction between the adapter protein S100A10 and its binding partner annexin A2 is a potentially important drug target. To obtain small-molecule starting points for inhibitors of this interaction, a three-dimensional pharmacophore model was constructed from the X-ray crystal structure of the complex between S100A10 and annexin A2. The pharmacophore model represents the favourable hydrophobic and hydrogen bond interactions between the two partners, as well as spatial and receptor site constraints (excluded volume spheres). Using this pharmacophore model, UNITY flex searches were carried out on a 3D library of 0.7 million commercially available compounds. This resulted in 568 hit compounds. Subsequently, GOLD docking studies were performed on these hits, and a set of 190 compounds were purchased and tested biochemically for inhibition of the protein interaction. Three compounds of similar chemical structure were identified as genuine inhibitors of the binding of annexin A2 to S100A10. The binding modes predicted by GOLD were in good agreement with their UNITY-generated conformations. We synthesised a series of analogues revealing areas critical for binding. Thus computational predictions and biochemical screening can be used successfully to derive novel chemical classes of protein–protein interaction blockers.

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N -Acyl phenylalanine analogues as potent small molecule VLA-4 antagonists

Cited by 21 publications

References 4 publications

A Cy5-Labeled S100A10 Tracer Used to Identify Inhibitors of the Protein Interaction With Annexin A2

A Cy5-Labeled S100A10 Tracer Used to Identify Inhibitors of the Protein Interaction With Annexin A2

The Identification and Optimization of Orally Efficacious, Small Molecule VLA-4 Antagonists

Three‐Dimensional Pharmacophore Design and Biochemical Screening Identifies Substituted 1,2,4‐Triazoles as Inhibitors of the Annexin A2–S100A10 Protein Interaction

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