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Principal Investigator: James Inglese
Organization: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
Fiscal Year: 2020
Award: $420,568
Funding agency: National Center for Advancing Translational Sciences
This project includes the development of biochemical and target-focused assays based on specific protein or nucleic acid targets implicated in disease. The assay designs are considered in the context of analysis and progression strategies for evaluation of a wide range of compound classes using high throughput screening technologies. There is a strong emphasis on methods development research to advance assay and lead discovery efficiency. Complementing these activities, we also explore and devise approaches for the interrogation of complex chemical libraries (e.g., natural product extracts, mRNA display). The work from this program is used to support a range of grant applications and prototype projects. The following are on-going:
Targeting protein palmitoylation with small molecules. In collaboration with A. Banerjee (NICHD, NIH) we have developed 1536-well compatible protein palmitoyl acyl transferase assays to evaluate chemical libraries for potential inhibitors of the enzyme as possible therapeutic leads for the large number of diseases to which this class of enzyme have been linked, including proteins associated with SARS CoV-2. These compounds are also anticipated to have value as structural, functional, and pharmacological probes.
Strategies for the discovery of small molecule ligands of RNA. In collaboration with Prof. N. Baird (U. Sciences) we are exploring assay designs to probe the interaction of small molecules with non-coding gene-regulatory mRNAs. The research has the potential of the discovery of novel antibiotics or anticancer agents.
Assay development to enable discovery of novel small molecule antagonists of the receptor guanylate cyclase Npr1. In collaboration with M. Hoon (NIDCR, NIH) we have developed assays of the b-type natriuretic peptide (BNP) receptor, Npr1. Recently the agonist, BNP was shown to be required for the transmission of itch sensation between peripheral and spinal cord nerves. The Npr1 assays were employed in large-scale chemical library screening to identify novel natriuretic peptide receptor antagonists to investigate the potential of pharmacological treatments of chronic itch, a condition that results in long-term unremitting urge to scratch that significantly degrades the quality of life for sufferers (Solinski HJ et al., 2019). This project is being continued under the HEAL initiative.
SIRPa-CD47 Protein-protein interaction. In collaboration with T. Miller (Paradigm Shift Therapeutics) and D. Roberts (NCI, NIH) our goal is to leverage the broad potential of CD47 as a molecular target in a number of tumors to create therapeutics that protect normal tissue from chemo and radiation therapy while differentially enhancing the effects of these therapies on the tumor. We designed and validated several biochemical SIRPa-CD47 protein-protein interaction (Miller T et al., 2019) and cellular assay formats (Burgess, T et al. 2020). Using these assays, we are investigating the potential SIRPa-CD47 complex disruption activity of several compounds identified from large-scale high throughput chemical library screening.
Chorismate mutase inhibitors. In collaboration with J. Padia (PrimeTime Life Sciences) this project seeks to develop a quantitative high throughput screening (qHTS) assay for the identification of small molecule inhibitors of chorismate mutase (CM). CM is an important enzyme found in plants and microorganisms required for the biosynthesis of the aromatic amino acids, phenylalanine, and tyrosine. Mammals cannot carry out the de novo biosynthesis of aromatic amino acids and must rely on dietary sources. Thus, a potent and selective drug-like inhibitor of CM would be a valuable antimicrobial agent, particularly for antimicrobial resistant infections.
Targeting G proteins with small molecules. Fibrous dysplasia of bone (McCune-Albright syndrome) is a hyperfunctioning endocrinopathy resulting from mis-sense mutations in the small -subunit of the G-protein, Gs leading to increased levels of cellular cAMP. The aim of this project is to develop biochemical and cell-based assays suitable for evaluating the activity and coupling of G proteins to their GPCRs and effector adenylyl cyclase (Getz RA et al., 2019). For example, by enabling a quantitative high throughput screening assay using the R201C mutant form of Gs to identify
small molecules capable of antagonizing the R201C Gs adenylyl cyclase-activating conformation.
Novel approaches to antifolate modulators of cell proliferation and immunosuppression. Methotrexate is a widely used chemotherapeutic for the treatment of cancer and rheumatoid arthritis. However, alternatives have been sought for decades due to the dose-limiting hematologic and hepatic toxicity of MTX, attributable in part to long-term cellular retention by polyglutamylation. This project is developing reduced folate utilizing enzyme inhibitors and assay methods to monitor cellular folate metabolism for more rapid evaluation of novel antifolates.
Terms: <2019 novel coronavirus><2019-nCoV><3'5'-cyclic ester of AMP><3,5 cyclic AMP synthetase><ANF receptor B><ANPB><Adenosine Cyclic 3',5'-Monophosphate><Adenosine Cyclic Monophosphate><Adenosine, cyclic 3',5'-(hydrogen phosphate)><Adenyl Cyclase><Adenylate Cyclase><Adenylyl Cyclase><Agonist><Albright Syndrome><Albright's syndrome><Anabolism><Anti-Cancer Agents><Antibiotic Agents><Antibiotic Drugs><Antibiotics><Antifolates><Antineoplastic Agents><Antineoplastic Drugs><Antineoplastics><Applications Grants><Aromatic Amino Acids><Assay><Atrial Natriuretic Peptide B-Type Receptor><Atrial Natriuretic Peptide Receptor B><Atrionatriuretic Peptide Receptor B><Atrophic Arthritis><B-type natriuretic peptide receptor><BNP Gene Product><BNP-32><Bioassay><Biochemical><Biologic Assays><Biologic Sciences><Biological Assay><Biological Sciences><Bioscience><Brain Natriuretic Peptide-32><Brain natriuretic peptide><CD47><CD47 Antigen><CD47 Glycoprotein><CD47 gene><Cancer Drug><Cancer Treatment><Cell Body><Cell Growth in Number><Cell Multiplication><Cell Proliferation><Cells><Cellular Assay><Cellular Proliferation><Chemicals><Chorismate Mutase><Chorismate Pyruvatemutase><Collaborations><Complement><Complement Proteins><Complex><Coupling><Cyclic AMP><Deoxyguanylate Cyclase><Development><Development and Research><Diet><Disease><Disorder><Dose-Limiting><Drugs><EXTMR><Enzyme Antagonist><Enzyme Gene><Enzyme Inhibitor><Enzyme Inhibitor Agent><Enzyme Inhibitor Drugs><Enzymes><Evaluation><Extramural><Extramural Activities><Fibrous Dysplasia of Bone><Folate><Folate Analog><Folate Inhibitors><Folate Metabolism><Folic Acid><Folic Acid Analog><Folic Acid Antagonists><Folic Acid Inhibitors><Folic Acid Metabolosm><Functional RNA><G Protein-Complex Receptor><G Protein-Coupled Receptor Genes><G-Protein-Coupled Receptors><G-Proteins><GC-B><GPCR><GTP-Binding Proteins><GTP-Regulatory Proteins><Goals><Grant Proposals><Guanine Nucleotide Coupling Protein><Guanine Nucleotide Regulatory Proteins><Guanyl Cyclase><Guanylate Cyclase><Guanylate Cyclase B><Hematology><Hepatotoxic effect><Hepatotoxicity><High Throughput Assay><Immunosuppression><Immunosuppression Effect><Immunosuppressive Effect><Inosinate Cyclase><Integrin-Associated Protein><Itching><Laboratories><Lead><Life Sciences><Ligands><Link><Liver Toxicity><MER6><MFR gene><MFR protein><MYD-1><Macrophage Fusion Receptor><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Mammalia><Mammals><McCune Albright's Syndrome><McCune-Albright Syndrome><Medication><Medulla Spinalis><Messenger RNA><Methods><Methotrexate><Methotrexate Methylaminopterin><Methotrexatum><Metotrexato><Miscellaneous Antibiotic><Missense Mutation><Molecular Configuration><Molecular Conformation><Molecular Stereochemistry><Molecular Target><Monitor><NICHD><NIDCR><NIDR><NIH><National Institute of Child Health and Human Development><National Institute of Children's Health and Human Development><National Institute of Dental Research><National Institute of Dental and Craniofacial Research><National Institutes of Health><Natriuretic Factor-32><Natriuretic Peptide Hormones><Natriuretic Peptide Receptor B><Natriuretic Peptides><Natural Products><Neoplastic Disease Chemotherapeutic Agents><Nerve><Nesiritide><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Normal Tissue><Normal tissue morphology><Nucleic Acids><Osteitis Fibrosa Disseminata><P84><PTPNS1><PTPNS1 gene><Pb element><Peptide Receptor><Peripheral><Pharmaceutic Preparations><Pharmaceutical Preparations><Pharmacological Treatment><Pharmacology><Phenylalanine><Plants><Protein-Tyrosine Phosphatase, Nonreceptor Type, Substrate 1><Proteins><Pruritic Disorder><Pruritis><Pruritus><Pteroylglutamic Acid><QOL><Quality of life><R & D><R&D><RNA><RNA Gene Products><Radiation therapy><Radiotherapeutics><Radiotherapy><Receptor Protein><Regulator Genes><Research><Rheumatoid Arthritis><Ribonucleic Acid><SARS-CoV-2><SARS-CoV2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related coronavirus 2><SHP Substrate 1><SHPS1><SIRP-Alpha-1><SIRPA><Science><Scientist><Severe acute respiratory syndrome coronavirus 2><Signal Regulatory Protein, Alpha Type, 1><Source><Spinal Cord><Structure><Surface Antigen Identified by Monoclonal Antibody 1D8><Technology><Therapeutic><Toxic effect on liver cells><Transcriptional Regulatory Elements><Transferase><Transferase Gene><Translations><Transmission><Tumor-Specific Treatment Agents><Type-B Natriuretic Peptide><Tyrosine><Tyrosine Phosphatase SHP Substrate 1><United States National Institutes of Health><Untranslated RNA><Vitamin M><Work><Wuhan coronavirus><adenosine 3'5' monophosphate><adenylcyclase><anti-cancer drug><anti-cancer therapy><anti-microbial agent><anti-microbial drug><anti-microbial resistant infection><anticancer agent><anticancer drug><anticancer therapy><antimicrobial agent><antimicrobial drug><antimicrobial resistant infection><assay development><atrial natriuretic factor receptor B><base><biosynthesis><brain Natriuretic factor><cAMP><cancer therapy><cell assay><chemical library><chronic itch><chronic pruritus><conformation><conformational state><design><designing><developmental><dietary><drug discovery><drug/agent><folate antagonist><folic acid metabolism><guanylyl cyclase><guanylyl cyclase-B receptor><heavy metal Pb><heavy metal lead><hepatic toxicity><hepatoxicity><high throughput screening><immune suppression><inhibitor><inhibitor/antagonist><itch sensation><mRNA><method development><microorganism><mutant><new approaches><noncoding><novel><novel approaches><novel strategies><novel strategy><palmitoylation><programs><protein protein interaction><prototype><radiation treatment><radio-therapy><receptor><regulatory gene><research and development><rheumatic arthritis><screening><small molecule><small molecule inhibitor><small molecule libraries><trans acting element><transmission process><treatment with radiation><tumor><vitamin Bc>