The Experts below are selected from a list of 20526 Experts worldwide ranked by ideXlab platform
Jeffrey W. Bode - One of the best experts on this subject based on the ideXlab platform.
-
Antibiotic Discovery with Synthetic Fermentation: Library Assembly, Phenotypic Screening, and Mechanism of Action of β-Peptides Targeting Penicillin-Binding Proteins
ACS chemical biology, 2019Co-Authors: Iain A. Stepek, Trung Cao, Anika Koetemann, Satomi Shimura, Bernd Wollscheid, Jeffrey W. BodeAbstract:In analogy to biosynthetic pathways leading to bioactive natural products, synthetic fermentation generates mixtures of molecules from simple building blocks under aqueous, biocompatible conditions, allowing the resulting cultures to be directly screened for biological activity. In this work, a novel β-peptide antibiotic was successfully identified using the synthetic fermentation platform. Phenotypic Screening was carried out in an initially random fashion, allowing simple identification of active cultures. Subsequent deconvolution, focused Screening, and structure–activity relationship studies led to the identification of a potent antimicrobial peptide, showing strong selectivity for our model system Bacillus subtilis over human HEK293 cells. To determine the antibacterial mechanism of action, a peptide probe bearing a photoaffinity tag was readily synthesized through the use of appropriate synthetic fermentation building blocks and utilized for target identification using a quantitative mass spectromet...
Seung Bum Park - One of the best experts on this subject based on the ideXlab platform.
-
Label-free target identification in drug discovery via Phenotypic Screening
Current opinion in chemical biology, 2019Co-Authors: Hankum Park, Seung Bum ParkAbstract:Phenotypic Screening has demonstrated its advantage in the discovery of first-in-class therapeutics, whereas target-based Screening has showed strength for follower drugs. Owing to the unbiased nature of Phenotypic Screening, novel druggable proteins can be uncovered by target identification. Chemical label-free target identification methods can eliminate the functionalization step of an original bioactive compound. Herein, we summarize recent advances in the development of label-free target identification methods, which are based on changes in protein stability against proteolysis, and chemical and thermal denaturation. Owing to the increasing application of shift in thermal stability for protein analysis in live cells and tissues, we mainly focus on the cellular stability shift assay and its proteome-wide application for target identification.
-
Phenotypic Screening to identify small molecule enhancers for glucose uptake target identification and rational optimization of their efficacy
Angewandte Chemie, 2014Co-Authors: Minseob Koh, Jongmin Park, Ja Young Koo, Donghyun Lim, Mi Young Cha, Jang Hyun Choi, Seung Bum ParkAbstract:Small-molecule glucose uptake enhancers targeted to myotubes and adipocytes were developed through a Phenotypic Screening linked with target identification and rational optimization. The target protein of glucose-uptake enhancers was identified as a nuclear receptor PPARγ (peroxisome proliferator-activated receptor gamma). Subsequent optimization of initial hits generated lead compounds with high potency for PPARγ transactivation and cellular glucose uptake. Finally, we confirmed that the chirality of optimized ligands differentiates their PPARγ transcriptional activity, binding affinity, and inhibitory activity toward Cdk5 (cyclin-dependent kinase 5)-mediated phosphorylation of PPARγ at Ser273. Using phenotype-based lead discovery along with early-stage target identification, this study has identified a new small-molecule enhancer of glucose uptake that targets PPARγ.
-
Phenotypic Screening to Identify Small‐Molecule Enhancers for Glucose Uptake: Target Identification and Rational Optimization of Their Efficacy
Angewandte Chemie (International ed. in English), 2014Co-Authors: Minseob Koh, Jongmin Park, Ja Young Koo, Donghyun Lim, Mi Young Cha, Jang Hyun Choi, Seung Bum ParkAbstract:Small-molecule glucose uptake enhancers targeted to myotubes and adipocytes were developed through a Phenotypic Screening linked with target identification and rational optimization. The target protein of glucose-uptake enhancers was identified as a nuclear receptor PPARγ (peroxisome proliferator-activated receptor gamma). Subsequent optimization of initial hits generated lead compounds with high potency for PPARγ transactivation and cellular glucose uptake. Finally, we confirmed that the chirality of optimized ligands differentiates their PPARγ transcriptional activity, binding affinity, and inhibitory activity toward Cdk5 (cyclin-dependent kinase 5)-mediated phosphorylation of PPARγ at Ser273. Using phenotype-based lead discovery along with early-stage target identification, this study has identified a new small-molecule enhancer of glucose uptake that targets PPARγ.
Iain A. Stepek - One of the best experts on this subject based on the ideXlab platform.
-
Antibiotic Discovery with Synthetic Fermentation: Library Assembly, Phenotypic Screening, and Mechanism of Action of β-Peptides Targeting Penicillin-Binding Proteins
ACS chemical biology, 2019Co-Authors: Iain A. Stepek, Trung Cao, Anika Koetemann, Satomi Shimura, Bernd Wollscheid, Jeffrey W. BodeAbstract:In analogy to biosynthetic pathways leading to bioactive natural products, synthetic fermentation generates mixtures of molecules from simple building blocks under aqueous, biocompatible conditions, allowing the resulting cultures to be directly screened for biological activity. In this work, a novel β-peptide antibiotic was successfully identified using the synthetic fermentation platform. Phenotypic Screening was carried out in an initially random fashion, allowing simple identification of active cultures. Subsequent deconvolution, focused Screening, and structure–activity relationship studies led to the identification of a potent antimicrobial peptide, showing strong selectivity for our model system Bacillus subtilis over human HEK293 cells. To determine the antibacterial mechanism of action, a peptide probe bearing a photoaffinity tag was readily synthesized through the use of appropriate synthetic fermentation building blocks and utilized for target identification using a quantitative mass spectromet...
-
Antibiotic Discovery with Synthetic Fermentation: Library Assembly, Phenotypic Screening, and Mechanism of Action of Beta-Peptides Targeting Penicillin-Binding Proteins
2019Co-Authors: Iain A. Stepek, Trung Cao, Anika Koetemann, Satomi Shimura, Bernd Wollscheid, Jeffrey BodeAbstract:In analogy to biosynthetic pathways leading to bioactive natural products, synthetic fermentation generates mixtures of molecules from simple building blocks under aqueous, biocompatible conditions, allowing for the resulting cultures to be directly screened for biological activity. In this work, a novel beta-peptide antibiotic was successfully identified using the synthetic fermentation platform. Phenotypic Screening was carried out in an initially random fashion, allowing for simple identification of active cultures. Subsequent deconvolution, focused Screening and structure-activity relationship studies led to the identification of a potent antimicrobial peptide, showing strong selectivity for our model system Bacillus subtilis over human Hek293 cells. To determine the antibacterial mechanism of action, a peptide probe bearing a photoaffinity tag was readily synthesized through the use of appropriate synthetic fermentation building blocks and utilized for target identification using a quantitative mass spectrometry-based strategy. The chemoproteomic approach led to the identification of a number of bacterial membrane proteins as prospective targets. These findings were validated through binding affinity studies with penicillin-binding protein 4 using microscale thermophoresis, with the bioactive peptide showing a dissociation constant (Kd) in the nanomolar range. Through these efforts, we provide a proof of concept for the synthetic fermentation approach presented here as a new strategy for the Phenotypic discovery of novel bioactive compounds.
Ho Jeong Kwon - One of the best experts on this subject based on the ideXlab platform.
-
Discovery of novel drug targets and their functions using Phenotypic Screening of natural products
Journal of Industrial Microbiology & Biotechnology, 2016Co-Authors: Junghwa Chang, Ho Jeong KwonAbstract:Natural products are valuable resources that provide a variety of bioactive compounds and natural pharmacophores in modern drug discovery. Discovery of biologically active natural products and unraveling their target proteins to understand their mode of action have always been critical hurdles for their development into clinical drugs. For effective discovery and development of bioactive natural products into novel therapeutic drugs, comprehensive Screening and identification of target proteins are indispensable. In this review, a systematic approach to understanding the mode of action of natural products isolated using Phenotypic Screening involving chemical proteomics-based target identification is introduced. This review highlights three natural products recently discovered via Phenotypic Screening, namely glucopiericidin A, ecumicin, and terpestacin, as representative case studies to revisit the pivotal role of natural products as powerful tools in discovering the novel functions and druggability of targets in biological systems and pathological diseases of interest.
-
Anti-Obesity Phenotypic Screening Looking to Increase OBR Cell Surface Expression
Journal of biomolecular screening, 2013Co-Authors: Tae-hee Kim, Ho Jeong Kwon, Dong Hwa Choi, Virginie Vauthier, Julie Dam, Yeon Ju Nam, Sang Hoon Shin, Jonathan CechettoAbstract:The leptin receptor, OBR, is involved in the regulation of whole-body energy homeostasis. Most obese people are resistant to leptin and do not respond to the hormone. The prevention and reversal of leptin resistance is one of the major current goals of obesity research. We showed previously that increased OBR cell surface expression concomitantly increases cellular leptin signaling and prevents obesity development in mice. Improvement of OBR cell surface expression can thus be considered as an interesting anti-obesity therapeutic strategy. To identify compounds that increase the surface expression of OBR, we developed a cell-based, Phenotypic assay to perform a high-content screen (HCS) against a library of 50,000 chemical compounds. We identified 67 compounds that increased OBR cell surface expression with AC 50 values in the low micromolar range and no effect on total OBR expression and cellular toxicity. Compounds were classified into 16 chemical clusters, of which 4 potentiated leptin-promoted signaling through the JAK2/STAT3 pathway. In conclusion, development of a robust Phenotypic Screening approach resulted in the discovery of four new scaffolds that demonstrate the desired biological activity and could constitute an original therapeutic solution against obesity and associated disorders.
-
Identification and validation of bioactive small molecule target through Phenotypic Screening.
Bioorganic & medicinal chemistry, 2011Co-Authors: Yoon Sun Cho, Ho Jeong KwonAbstract:For effective bioactive small molecule discovery and development into new therapeutic drug, a systematic Screening and target protein identification is required. Different from the conventional Screening system, herein Phenotypic Screening in combination with multi-omics-based target identification and validation (MOTIV) is introduced. First, Phenotypic Screening provides visual effect of bioactive small molecules in the cell or organism level. It is important to know the effect on the cell or organism level since small molecules affect not only a single target but the entire cellular mechanism within a cell or organism. Secondly, MOTIV provides systemic approach to discover the target protein of bioactive small molecule. With the chemical genomics and proteomics approach of target identification methods, various target protein candidates are identified. Then network analysis and validations of these candidates result in identifying the biologically relevant target protein and cellular mechanism. Overall, the combination of Phenotypic Screening and MOTIV will provide an effective approach to discover new bioactive small molecules and their target protein and mechanism identification.
Asier Unciti-broceta - One of the best experts on this subject based on the ideXlab platform.
-
Development of Potent Inhibitors of Receptor Tyrosine Kinases by Ligand-Based Drug Design and Target-Biased Phenotypic Screening.
Journal of medicinal chemistry, 2018Co-Authors: Samuel H. Myers, Douglas R. Houston, Carolin Temps, Valerie G. Brunton, Asier Unciti-brocetaAbstract:Pyrazolopyrimidines with potent antiproliferative properties were developed by an adaptive strategy that applies ligand-based design and Phenotypic Screening iteratively and is informed by biochemical assays. To drive development toward specific oncopathways, compounds were tested against cancer cells that overexpress, or not, AXL kinase. Identified Phenotypic hits were found to inhibit oncotargets AXL, RET, and FLT3. Subsequent optimization generated antiproliferative lead compounds with unique selectivity profiles, including selective AXL inhibitors and a highly potent inhibitor of FLT3.
-
Development of Potent Inhibitors of Receptor Tyrosine Kinases by Ligand-Based Drug Design and Target-Biased Phenotypic Screening
2018Co-Authors: Samuel H. Myers, Carolin Temps, Douglas R. Houston, Valerie G. Brunton, Asier Unciti-brocetaAbstract:Pyrazolopyrimidines with potent antiproliferative properties were developed by an adaptive strategy that applies ligand-based design and Phenotypic Screening iteratively and is informed by biochemical assays. To drive development toward specific oncopathways, compounds were tested against cancer cells that overexpress, or not, AXL kinase. Identified Phenotypic hits were found to inhibit oncotargets AXL, RET, and FLT3. Subsequent optimization generated antiproliferative lead compounds with unique selectivity profiles, including selective AXL inhibitors and a highly potent inhibitor of FLT3
-
Next-generation Phenotypic Screening.
Future medicinal chemistry, 2016Co-Authors: Scott J. Warchal, Asier Unciti-broceta, Neil O. CarragherAbstract:Phenotypic drug discovery (PDD) strategies are defined by Screening and selection of hit or lead compounds based on quantifiable Phenotypic endpoints without prior knowledge of the drug target. We outline the challenges associated with traditional Phenotypic Screening strategies and propose solutions and new opportunities to be gained by adopting modern PDD technologies. We highlight both historical and recent examples of approved drugs and new drug candidates discovered by modern Phenotypic Screening. Finally, we offer a prospective view of a new era of PDD underpinned by a wealth of technology advances in the areas of in vitro model development, high-content imaging and image informatics, mechanism-of-action profiling and target deconvolution.