A5000108912- Computational Drug Discovery Methods
- Innovative Microfluidic and Catalytic Techniques Innovation
- Antibiotic Resistance in Bacteria
- Analytical Chemistry and Chromatography
- Bacterial Genetics and Biotechnology
- Advanced biosensing and bioanalysis techniques
- Antibiotics Pharmacokinetics and Efficacy
- Biosimilars and Bioanalytical Methods
- Click Chemistry and Applications
- Cell Image Analysis Techniques
- Protein Degradation and Inhibitors
- RNA Interference and Gene Delivery
- Biotin and Related Studies
- Lipid Membrane Structure and Behavior
- Neurological diseases and metabolism
- Cellular transport and secretion
- Biochemical and Structural Characterization
- Axon Guidance and Neuronal Signaling
- Alzheimer's disease research and treatments
- Biochemical and Molecular Research
- Ubiquitin and proteasome pathways
- Bacteriophages and microbial interactions
- Wastewater Treatment and Nitrogen Removal
- Advanced Biosensing Techniques and Applications
- Yersinia bacterium, plague, ectoparasites research
University of Illinois Chicago
2025
University of California, Merced
2024
Enzo Life Sciences (United States)
2024
Novartis (United States)
2017-2022
Novartis Institutes for BioMedical Research
2019
Novartis (Switzerland)
2019
Novartis (Ireland)
2019
University of California, San Francisco
2003-2008
National Human Genome Research Institute
2007-2008
National Institutes of Health
2007-2008
Promiscuous small molecules plague screening libraries and hit lists. Previous work has found that several nonspecific compounds form submicrometer aggregates, it been suggested this aggregate species is responsible for the inhibition of many different enzymes. It not understood how aggregates inhibit their targets. To address question, biophysical, kinetic, microscopy methods were used to study interaction promiscuous, aggregate-forming inhibitors with model proteins. By use centrifugation...
High-throughput screening (HTS) is the primary technique for new lead identification in drug discovery and chemical biology. Unfortunately, it susceptible to false-positive hits. One common mechanism such false-positives congregation of organic molecules into colloidal aggregates, which nonspecifically inhibit enzymes. To both evaluate feasibility large-scale aggregate-based inhibition quantify its prevalence among hits, we tested 70,563 from National Institutes Health Chemical Genomics...
High-throughput screening (HTS) is widely used in drug discovery. Especially for screens of unbiased libraries, false positives can dominate "hit lists"; their origins are much debated. Here we determine the mechanism every active hit from a screen 70,563 molecules against β-lactamase using quantitative HTS (qHTS). Of 1274 initial inhibitors, 95% were detergent-sensitive and classified as aggregators. Among 70 remaining 25 potent, covalent-acting β-lactams. Mass spectra, counter-screens,...
The Hedgehog (Hh) pathway is essential for vertebrate embryogenesis, and excessive Hh target gene activation can cause cancer in humans. Here we show that Neuropilin 1 (Nrp1) Nrp2, transmembrane proteins with roles axon guidance vascular endothelial growth factor (VEGF) signaling, are important positive regulators of signal transduction. Nrps expressed at times locations active transduction during mouse development. Using cell lines lacking key components, mediate between activated...
Screening in mixtures is a common approach for increasing the efficiency of high-throughput screening. Here we investigate how "compound load" influences promiscuous aggregate-based inhibition. We screened 764 molecules individually and 10 at 5 μM each, comparing observed inhibition to that predicted from single-compound results. Synergistic effects on aggregation predominated, although antagonism was also observed. These results suggest screening can increase aggregation-based nonadditive manner.
In the preceding manuscript [Moreau et al. 2018, 10.1021/acs.jmedchem.7b01691] we described a successful fragment-based lead discovery (FBLD) strategy for of bacterial phosphopantetheine adenylyltransferase inhibitors (PPAT, CoaD). Following several rounds optimization two promising compounds were identified: triazolopyrimidinone 3 and 4-azabenzimidazole 4. Here disclose our efforts to further optimize these leads on-target potency Gram-negative cellular activity. Enabled by robust X-ray...
Secretory phospholipase A2 (sPLA2), an enzyme overexpressed in numerous diseases, has been used to trigger structural transformations lipid-based drug delivery systems, enabling payload release at target sites. Zwitterionic peptides are known for their superior antifouling properties, often outperforming poly(ethylene glycol) (PEG) surface modification by resisting protein adsorption. In this study, we examined lipid monolayers the water–vapor interface on a Langmuir trough, incorporating...
Objectives/Goals: Our aim was to identify how the epithelial–mesenchymal transition shields heterogeneous breast tumors against immune attack. Additionally, we endeavored understand whether our findings were conserved in canine mammary as a translational model for human tumors. Methods/Study Population: To interactions between quasi-mesenchymal (qM) tumor cells, epithelial (E) and cells within tumors, utilized preclinical mouse established lab. In this system, can precisely control...
Although immune checkpoint blockade therapy has generated dramatic responses in certain cancer types, breast tumors are largely unresponsive. Epithelial-mesenchymal plasticity leads to the assembly of an immunosuppressive tumor microenvironment and drives resistance immunotherapies. Importantly, targeting CD73 completely sensitizes quasi-mesenchymal anti-CTLA4 therapy. However, mechanism(s) sensitization remained unknown. We demonstrate that them a CD4 + T-cell dependent manner. Moreover,...
The discovery and development of new antibiotics capable curing infections due to multidrug-resistant pandrug-resistant Gram-negative bacteria are a major challenge with fundamental importance our global healthcare system. Part broad program at Novartis address this urgent, unmet need includes the search for agents that inhibit novel bacterial targets. Here we report hit-to-lead optimization inhibitors phosphopantetheine adenylyltransferase (PPAT) from bacteria. Utilizing fragment-based...
Beta-lactams comprise one of the earliest classes antibiotic therapies. These molecules covalently inhibit enzymes from family penicillin-binding proteins (PBPs), which are essential in construction bacterial cell wall. As a result, beta-lactams cause striking changes to cellular morphology, nature varies by range PBPs simultaneously engaged cell. The traditional method exploring beta-lactam polyspecificity is gel-based binding assay low-throughput and typically run ex situ extracts. Here,...
The Gram-negative cell envelope presents a formidable barrier to xenobiotics, and achieving sufficient compound exposure inside the is key challenge for discovery of new antibiotics. To provide insight on molecular determinants governing in bacteria, we developed methodology leveraging cyclooctyne-based bioorthogonal probe assess compartment-specific exposure. This can be selectively localized periplasmic or cytoplasmic compartments bacteria. Once localized, used test azide-containing...
Drug-resistant Gram-negative bacteria are of increasing concern worldwide. Novel antibiotics needed, but their development is complicated by the requirement to simultaneously optimize molecules for target affinity and cellular potency, which can result in divergent structure-activity relationships (SARs). These challenges were exemplified during our attempts inhibitors bacterial enzyme CoaD originally identified through a biochemical screen. To facilitate lead optimization, we developed mass...
Compartmentalization is a crucial facet of many biological systems, and key aspects cellular processes rely on spatial segregation within the cell. While drug targets reside in specific intracellular compartments, tools available for assessing compound exposure are generally limited to whole-cell measurements. To address this gap, we recently developed bioorthogonal chemistry-based method assess compartment-specific demonstrated its use Gram-negative bacteria. expand applicability approach,...
Combination therapies are common in many therapeutic contexts, including infectious diseases and cancer. A approach for evaluating combinations vitro is to assess effects on cell growth as synergistic, antagonistic, or neutral using "checkerboard" experiments systematically sample of agents multiple doses. To further understand the antibiotic combinations, we employed high-content imaging study morphological changes caused by combination treatments checkerboard experiments. Using an...
Lipopolysacharride (LPS) forms the outer leaflet of membrane in Gram-negative bacteria and contributes to permeability barrier immune response. In this study, we established a method for monitoring LPS biosynthetic intermediates Raetz pathway (lpxA-lpxK) Escherichia coli. Metabolites from compound-treated cells genetically-perturbed were extracted whole concentrated by mixed-mode weak anion exchange (WAX) solid-phase extraction (SPE) prior analysis normal phase (NP)LC-MS/MS. Data was...
<title>Abstract</title> The discovery of bifunctional degradation activating compounds (BiDACs) has led to the development a new class drugs that promote clearance their protein targets. BiDAC-induced ubiquitination is generally believed direct cytosolic and nuclear proteins proteolytic destruction by proteasomes. However, pathways govern other classes BiDAC targets, such as integral membrane intraorganellar proteins, have not been investigated in depth. In this study we used morphological...
ADVERTISEMENT RETURN TO ISSUEPREVAddition/CorrectionNEXTSynergy and Antagonism of Promiscuous Inhibition in Multiple-Compound MixturesBrian Y. Feng Brian K. ShoichetCite this: J. Med. Chem. 2007, 50, 12, 2930Publication Date (Web):May 18, 2007Publication History Published online18 May 2007Published inissue 1 June 2007https://pubs.acs.org/doi/10.1021/jm0703439https://doi.org/10.1021/jm0703439correctionACS PublicationsCopyright © 2007 American Chemical Society. This publication is available...