Article
Chemistry, Medicinal
Jon Kyle Awalt, Anh T. N. Nguyen, Tim J. Fyfe, Bui San Thai, Paul J. White, Arthur Christopoulos, Manuela Joerg, Lauren T. May, Peter J. Scammells
Summary: The adenosine A(1) receptor is a therapeutic target for cardioprotection during myocardial ischemia and reperfusion injury. However, clinical translation of A(1)R agonists is hindered by dose-limiting adverse effects. The bitopic agonist VCP746, consisting of an adenosine pharmacophore linked to an allosteric moiety, has shown promising results in stimulating cardioprotective A(1)R signaling effects without unwanted bradycardia. This study investigates the structure-activity relationships of VCP746 and identifies the most potent A2BR agonist to date.
JOURNAL OF MEDICINAL CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Min Guo, Abolghasem Bakhoda, Zhan-Guo Gao, Joseph M. Ramsey, Yang Li, Kelly A. O'Conor, Andrew C. Kelleher, Seth M. Eisenberg, Yeona Kang, Xuefeng Yan, Cameron Javdan, Joanna S. Fowler, Kenner C. Rice, Jacob M. Hooker, Kenneth A. Jacobson, Sung Won Kim, Nora D. Volkow
Summary: Efforts to develop full agonist adenosine A(1) radioligands for PET imaging showed promising binding affinity and selectivity, but poor brain permeability in rodent models potentially due to being substrates for brain efflux pumps. Computational evaluation suggested that high molecular weight and high polar surface area were main factors contributing to low brain penetration of these compounds. These findings provide valuable insights for future development of highly potent CNS A(1)AR drugs.
ACS CHEMICAL NEUROSCIENCE
(2021)
Article
Immunology
Liqing Chen, Xiaoxiao Qi, Dan Liang, Guiqi Li, Xiaofang Peng, Xiaohui Li, Bixia Ke, Huanying Zheng, Zhongqiu Liu, Changwen Ke, Guochao Liao, Liang Liu, Qian Feng
Summary: This study provides valuable information for the ongoing development of recombinant protein-based SARS-CoV-2 vaccines and serves as a basis for booster vaccinations to prevent reinfection with SARS-CoV-2 variants.
Article
Chemistry, Medicinal
Dilip K. Tosh, Courtney L. Fisher, Veronica Salmaso, Tina C. Wan, Ryan G. Campbell, Eric Chen, Zhan-Guo Gao, John A. Auchampach, Kenneth A. Jacobson
Summary: (N)-Methanocarba adenosine derivatives were modified to form macrocyclic A(3) adenosine receptor agonists. These macrocycles retained affinity and had a spatially proximal orientation on the receptor. C2-Arylethynyl-linked macrocycle 19 showed higher selectivity for A(3) adenosine receptor compared to 2-ether-linked macrocycle 12.
ACS PHARMACOLOGY & TRANSLATIONAL SCIENCE
(2023)
Meeting Abstract
Cardiac & Cardiovascular Systems
P. Pannucci, J. March, S. L. Cooper, S. J. Hill, J. Woolard
CARDIOVASCULAR RESEARCH
(2022)
Article
Chemistry, Medicinal
Zhi Yuan Kok, Leigh A. Stoddart, Sarah J. Mistry, Tamara A. M. Mocking, Henry F. Vischer, Rob Leurs, Stephen J. Hill, Shailesh N. Mistry, Barrie Kellam
Summary: This study reports a series of high-affinity H1R fluorescent ligands, which overcome the limitations of linker SAR by incorporating a BODIPY 630/650-based fluorophore, and successfully visualizes HA at a concentration of 10 nM using confocal microscopy.
JOURNAL OF MEDICINAL CHEMISTRY
(2022)
Article
Pharmacology & Pharmacy
Sean A. Cullum, Dmitry B. Veprintsev, Stephen J. Hill
Summary: This study analysed the kinetic phases of cAMP responses to beta(2)-adrenoceptor agonists in HEK293 cells expressing low levels of endogenous beta(2)-adrenoceptor. It was found that partial agonists had reduced relative IRmax values compared to isoprenaline. Preincubation with beta(2)-adrenoceptor antagonists led to a decrease in the peak response and exacerbated the effect on IRmax values. These findings provide insights into the hemi-equilibria in low receptor reserve systems with agonist-antagonist interactions.
BRITISH JOURNAL OF PHARMACOLOGY
(2023)
Article
Pharmacology & Pharmacy
Charles S. Lay, Laura E. Kilpatrick, Peter D. Craggs, Stephen J. Hill
Summary: In this study, the signalling mechanism of IL-23 receptor complexes was explored using NanoBiT and BRET techniques. The results showed that IL-23 receptor complexes formed to the same degree with and without ligand, and signal transduction was induced upon ligand binding. These findings have important implications for drug development targeting the IL-23 receptor.
BRITISH JOURNAL OF PHARMACOLOGY
(2023)
Article
Immunology
Dehan Comez, Jacqueline Glenn, Stephanie M. M. Anbuhl, Raimond Heukers, Martine J. J. Smit, Stephen J. J. Hill, Laura E. E. Kilpatrick
Summary: In this study, NanoBRET technique was used to compare the binding characteristics of two distinct EGFR targeting nanobodies to full length EGFR. The results showed that these nanobodies had different functions, with one inhibiting EGF binding to EGFR and the other acting as a conformational sensor for agonist bound EGFR.
FRONTIERS IN IMMUNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Jak Grimes, Zsombor Koszegi, Yann Lanoiselee, Tamara Miljus, Shannon L. O'Brien, Tomasz M. Stepniewski, Brian Medel-Lacruz, Mithu Baidya, Maria Makarova, Ravi Mistry, Joelle Goulding, Julia Drube, Carsten Hoffmann, Dylan M. Owen, Arun K. Shukla, Jana Selent, Stephen J. Hill, Davide Calebiro
Summary: 3-arrestin plays a key role in GPCR signaling and desensitization. This study combines single-molecule microscopy and molecular dynamics simulations to investigate the interactions between 3-arrestin, receptors, and the lipid bilayer. Surprisingly, the results show that 3-arrestin spontaneously inserts into the lipid bilayer and interacts with receptors through lateral diffusion on the plasma membrane. Furthermore, the plasma membrane stabilizes 3-arrestin after receptor interaction, allowing it to separate from the activating receptor and diffuse to clathrin-coated pits. These findings provide new insights into the mechanism of 3-arrestin function at the plasma membrane.
Review
Biochemistry & Molecular Biology
Patrizia Pannucci, Sophie R. Jefferson, Jonathan Hampshire, Samantha L. Cooper, Stephen J. Hill, Jeanette Woolard
Summary: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection responsible for COVID-19 commonly leads to dyspnea and fatigue, primarily affecting the lungs. However, it can also result in dysfunction of extra-pulmonary organs, particularly the cardiovascular system. Cardiac complications such as hypertension, thromboembolism, arrhythmia, and heart failure have been observed in COVID-19 patients, with myocardial injury and myocarditis being the most frequent. Understanding the mechanisms of COVID-19-induced myocarditis, including changes in ACE2 expression and immune responses, is crucial.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Chemistry, Medicinal
Sebastian Dekkers, Birgit Caspar, Joelle Goulding, Nicholas D. Kindon, Laura E. Kilpatrick, Leigh A. Stoddart, Stephen J. Briddon, Barrie Kellam, Stephen J. Hill, Michael J. Stocks
Summary: In this study, fluorescent probes based on previously reported small-molecule antagonists were designed and synthesized using classic medicinal chemistry approaches to investigate the pharmacology and cellular distribution of the CXCR4 receptor. Three distinct chemical classes of fluorescent probes were developed and shown to specifically bind to the CXCR4 receptor in a fluorescence-based NanoBRET binding assay (pKD ranging 6.6-7.1). These probes were also used in competition binding experiments and confocal microscopy to further explore the pharmacology and cellular distribution of CXCR4.
JOURNAL OF MEDICINAL CHEMISTRY
(2023)
Article
Multidisciplinary Sciences
Charles S. Lay, Albert Isidro-Llobet, Laura E. Kilpatrick, Peter D. Craggs, Stephen J. Hill
Summary: Using a fluorescent version of IL-23, this study characterized antagonists of the full-length receptor expressed by living cells and developed a cyclic peptide fluorescent probe specific to the IL23p19:IL23R interface to further study receptor antagonists. The study also demonstrated that the immunocompromising C115Y IL23R mutation disrupts the binding epitope for IL23p19.
NATURE COMMUNICATIONS
(2023)
Article
Pharmacology & Pharmacy
Marieke Van Daele, Laura E. Kilpatrick, Jeanette Woolard, Stephen J. Hill
Summary: Vascular endothelial growth factor (VEGF) is crucial for angiogenesis and vascular endothelial cell functions. The study developed a ligand-binding assay to monitor the binding affinity and kinetics of different tyrosine kinase inhibitors (TKIs) to VEGF receptor 2 (VEGFR2). The results showed that compounds inhibiting the binding of a fluorescent analogue of sunitinib (sunitinib-red) also attenuated VEGFR2-mediated signaling, suggesting potential cardiovascular liabilities.
BIOCHEMICAL PHARMACOLOGY
(2023)
Review
Pharmacology & Pharmacy
Stephen J. Hill, Laura E. Kilpatrick
Summary: Equilibrium binding assays are commonly used in drug discovery to evaluate drug-receptor interactions, but there is increasing interest in studying the kinetics of these interactions. Drugs can induce conformational changes in the orthosteric binding site, leading to alterations in the association and dissociation rates of orthosteric ligands. This review provides an overview of how fluorescent ligand technologies are used to study ligand-receptor kinetics in living cells and the insights they offer into conformational changes induced by drugs targeting cell surface receptors.
BRITISH JOURNAL OF PHARMACOLOGY
(2023)
Article
Multidisciplinary Sciences
Lydia Ogrodzinski, Simon Platt, Joelle Goulding, Cameron Alexander, Tracy D. Farr, Jeanette Woolard, Stephen J. Hill, Laura E. Kilpatrick
Summary: E-selectin is a protein expressed on endothelial cells in response to inflammatory cytokines, and it plays a role in leukocyte rolling and extravasation. This study used CRISPR-Cas9 genome editing and NanoLuc Binary Technology (NanoBiT) to tag endogenous E-selectin in human umbilical vein endothelial cells (HUVECs) and monitor its expression in real time. The combination of NanoBiT and CRISPR-Cas9 gene editing provides a powerful tool for monitoring dynamic changes in E-selectin expression on cell surfaces, which can contribute to the discovery of drugs targeting this important inflammatory protein.
Article
Hematology
Foteini-Nafsika Damaskinaki, Natalie J. Jooss, Eleyna M. Martin, Joanne C. Clark, Mark R. Thomas, Natalie S. Poulter, Jonas Emsley, Barrie Kellam, Steve P. Watson, Alexandre Slater
Summary: This study investigates the binding sites of three high-affinity nanobodies, Nb2, Nb21, and Nb35, on the platelet-signaling receptor GPVI. The researchers found that all three nanobodies can bind to the D1 domain of GPVI and inhibit collagen-induced GPVI signaling. They also identified common target residues, Arg46, Tyr47, and Ala57, on GPVI for these nanobodies. Additionally, the study negates the idea that GPVI dimerization induces a conformational change required for ligand binding.
JOURNAL OF THROMBOSIS AND HAEMOSTASIS
(2023)
Article
Materials Science, Biomaterials
Alessandra Travanut, Patricia F. Monteiro, Sean Smith, Steven M. Howdle, Anna M. Grabowska, Barrie Kellam, Michael A. R. Meier, Cameron Alexander
Summary: Passerini reactions are suitable for the preparation of drug delivery materials, enabling efficient drug conjugation and encapsulation, as well as enhanced efficacy against TNBC cells.
JOURNAL OF MATERIALS CHEMISTRY B
(2022)
