Ligand identification for G-protein-coupled receptors: a lead generation perspective

Bleicher, Konrad H.; Green, Luke G.; Martin, Rainer E.; Rogers‐Evans, Mark

doi:10.1016/j.cbpa.2004.04.008

Cited by 40 publications

(44 citation statements)

References 62 publications

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“…[75][76][77] Generally, the design of deorphanization libraries can be distinguished from targeted lead-finding libraries. Given the broad chemical diversity of the hormones that are recognized by GPCRs, deorphanization libraries try to cover as many known active chemical classes as possible.…”

Section: Designing Compound Libraries Targeting Gpcrsmentioning

confidence: 99%

The 7 TM G‐Protein‐Coupled Receptor Target Family

et al. 2006

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Chemical biology approaches have a long history in the exploration of the G-protein-coupled receptor (GPCR) family, which represents the largest and most important group of targets for therapeutics. The analysis of the human genome revealed a significant number of new members with unknown physiological function which are today the focus of many reverse pharmacology drug-discovery programs. As the seven hydrophobic transmembrane segments are a defining common structural feature of these receptors, and as signaling through heterotrimeric G proteins is not demonstrated in all cases, these proteins are also referred to as seven transmembrane (7 TM) or serpentine receptors. This review summarizes important historic milestones of GPCR research, from the beginning, when pharmacology was mainly descriptive, to the age of modern molecular biology, with the cloning of the first receptor and now the availability of the entire human GPCR repertoire at the sequence and protein level. It shows how GPCR-directed drug discovery was initially based on the careful testing of a few specifically made chemical compounds and is today pursued with modern drug-discovery approaches, including combinatorial library design, structural biology, molecular informatics, and advanced screening technologies for the identification of new compounds that activate or inhibit GPCRs specifically. Such compounds, in conjunction with other new technologies, allow us to study the role of receptors in physiology and medicine, and will hopefully result in novel therapies. We also outline how basic research on the signaling and regulatory mechanisms of GPCRs is advancing, leading to the discovery of new GPCR-interacting proteins and thus opening new perspectives for drug development. Practical examples from GPCR expression studies, HTS (high-throughput screening), and the design of monoamine-related GPCR-focused combinatorial libraries illustrate ongoing GPCR chemical biology research. Finally, we outline future progress that may relate today's discoveries to the development of new medicines.

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Section: Designing Compound Libraries Targeting Gpcrsmentioning

confidence: 99%

The 7 TM G‐Protein‐Coupled Receptor Target Family

et al. 2006

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“…For example, this motif is found in potent and selective EP3 antagonists developed by Merck and dual NK1/NK2 antagonist developed by Novartis. 3 This biphenyl fragment has been found within the top 10 antagonistic scaffolds such as B3 or B8. The common pharmacophoric groups of non-peptide human urotensin-II antagonists ( Figure 5) are hydrogen-bond acceptor and donor, ionizable group (basic amine), and aromatic hydrophobic features.…”

Section: Resultsmentioning

confidence: 99%

“…Among GPCR antagonistic features found from our analysis, fragments like sulfonamide group and benzodiazepine-like scaffold are usually found in clinically used antagonists. 3 GPCRs are often the first ranking for medicinal chemists concerning the druggability of a target. The present study is a ligand-based retrospective analysis of the classification of GPCR agonistic as well as antagonistic activities using a Bayesian statistical approach.…”

Section: Resultsmentioning

confidence: 99%

“…1,2 In addition to their widespread appearance and highly complex function in nature, GPCRs are modulated by a plethora of diverse endogenous and exogenous ligand. 3 In fact, several ligands for GPCRs are found among the worldwide top-100-selling pharmaceutical products. 1 It is believed that a receptor molecule exits in a conformational equilibrium between active and inactive biophysical state.…”

Section: Introductionmentioning

confidence: 99%

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Bayesian Model for the Classification of GPCR Agonists and Antagonists

Choi¹,

Kim²,

Jung³

et al. 2010

Bulletin of the Korean Chemical Society

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G-protein coupled receptors (GPCRs) are involved in a wide variety of physiological processes and are known to be targets for nearly 50% of drugs. The various functions of GPCRs are affected by their cognate ligands which are mainly classified as agonists and antagonists. The purpose of this study is to develop a Bayesian classification model, that can predict a compound as either human GPCR agonist or antagonist. Total 6627 compounds experimentally determined as either GPCR agonists or antagonists covering all the classes of GPCRs were gathered to comprise the dataset. This model distinguishes GPCR agonists from GPCR antagonists by using chemical fingerprint, FCFP_6. The model revealed distinctive structural characteristics between agonistic and antagonistic compounds: in general, 1) GPCR agonists were flexible and had aliphatic amines, and 2) GPCR antagonists had planar groups and aromatic amines. This model showed very good discriminative ability in general, with pretty good discriminant statistics for the training set (accuracy: 90.1%) and a good predictive ability for the test set (accuracy: 89.2%). Also, receiver operating characteristic (ROC) plot showed the area under the curve (AUC) to be 0.957, and Matthew's Correlation Coefficient (MCC) value was 0.803. The quality of our model suggests that it could aid to classify the compounds as either GPCR agonists or antagonists, especially in the early stages of the drug discovery process.

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“…25 The CXC chemokine receptor-2 (CXCR2), a member of the large Family A type GPCRs, is involved in many ailments, including cancers and inflammatory diseases, and it is therefore an important target for drug discovery. 1,[5][6][7][8][9]16,[26][27][28] A diversity of therapeutics is used to inhibit the receptor signaling function of a number of GPCRs, e.g., small molecules, 29,30 peptide analogs of the natural ligands, 31,32 or monoclonal antibodies; 16 however, no monoclonal antibody therapeutics targeting CXCR2 have been clinically developed so far.…”

Section: Discussionmentioning

confidence: 99%

A combination of in vitro techniques for efficient discovery of functional monoclonal antibodies against human CXC chemokine receptor-2 (CXCR2)

Boshuizen

Marsden

Turkstra

et al. 2014

mAbs

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y equal contribution # Ronald S Boshuizen is currently at Astellas Pharma Europe BV; Meppel, The Netherlands; Christine J Rossant is currently at Crescendo Biologics; Cambridge, United Kingdom; Klaus Schwamborn is currently at VALNEVA SE; Nantes, France Keywords: GPCR, CXCR2, monoclonal antibody, phage display library, ligand inhibition Abbreviations: ABTS, 2,2 0 -azino-bis(3-ethylbenzothiazoline-6-sulphonic acid); BSA, bovine serum albumin; CLIPS TM , Chemical LInkage of Peptides onto Scaffolds; ECL, extracellular loop; EDTA, ethylenediaminetetraacetic acid; ELISA, enzyme-linked immunoabsorbent assay; Fmoc, fluorenylmethyloxycarbonyl; GPCR, G-protein coupled receptor; Gro-a, growth-related protein a; IL-8, interleukin 8; IPTG, isopropyl b-D-1-thiogalactopyranoside; MFI, mean fluorescence intensity; PBS, phosphate buffer saline; PCR, polymerase chain reaction; PEG, polyethyleneglycol; scFv, single-chain variable fragment; TES, 2-propan -2-yl]amino]ethanesulfonic acid; TRIS, tris(hydroxymethyl)aminomethaneBackground: Development of functional monoclonal antibodies against intractable GPCR targets.Results: Identification of structured peptides mimicking the ligand binding site, their use in panning to enrich for a population of binders, and the subsequent challenge of this population with receptor overexpressing cells leads to functional monoclonal antibodies.Conclusion: The combination of techniques provides a successful strategic approach for the development of functional monoclonal antibodies against CXCR2 in a relatively small campaign.Significance: The presented combination of techniques might be applicable for other, notoriously difficult, GPCR targets.Summary: The CXC chemokine receptor-2 (CXCR2) is a member of the large 'family A' of G-protein-coupledreceptors and is overexpressed in various types of cancer cells. CXCR2 is activated by binding of a number of ligands, including interleukin 8 (IL-8) and growth-related protein a (Gro-a). Monoclonal antibodies capable of blocking the ligand-receptor interaction are therefore of therapeutic interest; however, the development of biological active antibodies against highly structured GPCR proteins is challenging. Here we present a combination of techniques that improve the discovery of functional monoclonal antibodies against the native CXCR2 receptor.The IL-8 binding site of CXCR2 was identified by screening peptide libraries with the IL-8 ligand, and then reconstructed as soluble synthetic peptides. These peptides were used as antigens to probe an antibody fragment phage display library to obtain subpopulations binding to the IL-8 binding site of CXCR2. Further enrichment of the phage population was achieved by an additional selection round with CXCR2 overexpressing cells as a different antigen source. The scFvs from the CXCR2 specific phage clones were sequenced and converted into monoclonal antibodies. The obtained antibodies bound specifically to CXCR2 expressing cells and inhibited the IL-8 and Gro-a induced ß-arrestin recruitment with IC50 values of 0.3 an...

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Ligand identification for G-protein-coupled receptors: a lead generation perspective

Cited by 40 publications

References 62 publications

The 7 TM G‐Protein‐Coupled Receptor Target Family

The 7 TM G‐Protein‐Coupled Receptor Target Family

Bayesian Model for the Classification of GPCR Agonists and Antagonists

A combination of in vitro techniques for efficient discovery of functional monoclonal antibodies against human CXC chemokine receptor-2 (CXCR2)

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