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A Genomic Approach to Discovering Novel Cathepsin Inhibitors from Cyanobacteria

从蓝藻中发现新型组织蛋白酶抑制剂的基因组方法

基本信息

批准号：
10388862
负责人：
Nicole Elizabeth Avalon
金额：
$ 6.93万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10388862
关键词：
Active Sites Anabaena Anabolism Aneurysmal Subarachnoid Hemorrhages Animal Model Antibiotic Resistance Architecture Atlases Behavioral Bioinformatics Biological Assay Biology Bypass Caspase Cathepsin L Cathepsins Cathepsins B Cells Cerebral Ischemia Cerebral hemisphere hemorrhage Cessation of life Chemicals Collection Complex Crystallization Cyanobacterium Data Docking Drug Targeting Environmental Impact Enzymes FDA approved Fellowship Gene Cluster Gene Expression Genome Genomic Library Genomic approach Genomics Goals In Vitro Injury Institution Ion Channel Ischemia Knock-out Lead Libraries Literature Methods Mining Modeling Molecular Morbidity - disease rate Mus National Research Service Awards Natural Products Neurocognitive Neurocognitive Deficit Neurologic Neurological outcome Neuronal Injury Neurons Oceanography Organism Outcome Pathway interactions Pattern Peptide Hydrolases Peptides Permeability Pharmaceutical Preparations Pharmacologic Substance Pharmacology Phylogeny Plasmids Postdoctoral Fellow Protease Inhibitor Proteins Reporter Ribosomes Scheme Secondary to Seeds Source Specificity Structure Techniques Technology Testing Traumatic Brain Injury Tubulin United States United States National Institutes of Health analog base behavioral outcome clinical development design drug candidate drug discovery experimental study improved in silico inhibitor innovation interdisciplinary approach interest markov model mortality nanomolar neuroinflammation neuropathology novel novel therapeutics overexpression peptide synthase pharmacophore polyketide synthase post-doctoral training pre-clinical prevent professor prognostic resistance gene small molecule synergism tandem mass spectrometry tool virtual yeast two hybrid system

项目摘要

Project Abstract Traumatic brain injury (TBI) is a leading cause of morbidity and mortality world-wide and in the United States;1,2 however, there are no FDA-approved medications to ameliorate the devastating consequences of secondary injury caused by complex neuropathological cascades following the initial neuronal insult. Cathepsin B and L represent promising drug targets, as knock-out and inhibition studies of these cysteine proteases in animal models reveal improvement in behavioral and neurocognitive sequelae.3–7 There is a clear need for new therapeutic options to treat secondary injury following TBI, and cathepsin B and L inhibitors have pharmaceutical promise. Over 60% of FDA-approved drugs are derived from or inspired by natural products (NPs).8 Cyanobacteria are known to produce NPs with a range of bioactivity, including activity as protease inhibitors.9,10 NPs are biosynthesized by megaenzymes that are encoded as discrete genomic packages called biosynthetic gene clusters (BGCs).11 We hypothesize that a genomic approach can be used to enhance drug discovery efforts from cyanobacteria for novel cathepsin B and L inhibitors through the three aims outlined below. First, an innovative pharmacophore-based genome mining pipeline will be developed (Aim 1). In this aim, pharmacophores deduced from virtual docking experiments and known cathepsin inhibitors will be used to predict enzymatic domains responsible for the creation of the desired pharmacophore. This retrobiosynthetic prediction will be used to make bioinformatic models to interrogate sequenced cyanobacterial genomes to find candidate BGCs. The BGCs of highest interest will be identified, delineated, and the compounds produced through either cultivation or heterologous expression (Aim 2). Using a full retrobiosynthetic prediction for gallinamide A, a compound that demonstrates nanomolar cathepsin L inhibition, we will search for the BGC within the genomic library at Scripps or in new cyanobacterial collections. Additional BGCs from Aim 1 will be developed in this aim as well. If the BGC is not constitutively expressed or if the BGC is not associated with a cultivatable organism, heterologous expression will be pursued (Aim 3). Compound isolation and molecular networking to analyze and annotate the chemical space of the natural products produced will follow. Compounds will be tested in bioassays to evaluate cathepsin B and L activity on purified enzymes as well as in neuronal and glial cellular studies. The gap between the identified pharmaceutical need and the discovery of novel bioactive molecules can be bridged by the innovative multidisciplinary approach presented in this application. The proposed project will be carried out as part of an NIH F32 NRSA Fellowship at Scripps Institution of Oceanography, UC San Diego, under the co-sponsorship of Professors William Gerwick and Vivian Hook and a team of collaborators that will train the postdoctoral fellow in virtual docking experiments, bioinformatics, heterologous gene expression, and cathepsin-related bioassays.

项目摘要