Jon S. Thorson - US grants

DESCRIPTION (provided by applicant): Roughly 70 percent of current lead compounds in modern drug discovery derive directly from the natural products, many of which are glycosylated bacterial metabolites. While it is known that the sugar ligands of these pharmaceutically important metabolites often define their corresponding biological activity, efficient methods to alter these essential carbohydrate ligands are still lacking. This proposal outlines a stepwise approach to accomplish this goal while also providing invaluable mechanistic and structural information on two critical, but poorly understood, enzyme classes; namely, nucleotidylytransferases and glycosyltransferases. Specifically, the proposed studies are designed to exploit structure/function-based protein engineering to generate a promiscuous in vitro nucleotidylyltransferase/glycosyltransferase systems which will provide a library of potentially new bioactive metabolites. The model system selected includes the Salmonella rmlA-encoded alpha-D-glucose-1-phosphate thymidylyltansferase (Ep) and three glycosyltransferases, which act ponerthronolide B (EryBV, EryCIII and MegGT). Among the many advantages of the presented model, it has been shown that the MegGT-catalyzed addition of a single sugar (megosamine) to erthromycin leads to a metabolite (megalomicin) with remarkably different biological activity. Thus, the selected model promises varied biological activity from metabolites anticipated from the proposed studies. The specific goals include: 1) structural and mechanistic studies on Ep3 EryBV, EryCIII and MegGT, 2) structure-based Ep engineering for the construction of nucleotide sugar libraries and 3) the use of this library, in conjunction with EryBV, EryCIII and MegGT, to generate libraries of glycosylated erythronolides with potentially new bioactivity.

DESCRIPTION (provided by applicant): Calicheamicin gamma1 from Micromonospora echinospora spp. calichensis is the most prominent of the 10- membered enediyne family with respect to its unprecedented molecular architecture, spectacular biological activity and clinical value. As such, calicheamicin is an excellent target for the study of natural product biosynthesis and self-resistance. The objective of the first phase of this study was to i) pursue the biosynthesis of the DNA-delivery component of calicheamicin (the aryltetrasaccharide, comprised of four uniquely functionalized sugars), ii) develop the genetic tools (transformation and gene disruption protocols) to address calicheamicin biosynthesis in Micromonospora and iii) investigate the mechanism(s) of calicheamicin self-resistance in Micromonospora. With these goals achieved and new tools/information in place, the second phase of this massive project will predominately focus upon expanding this program toward understanding and exploiting the complex biosynthesis of the enediyne core. While continuing our focus upon calicheamicin as a model for 10-membered enediyne biosynthesis, a second complimentary 10-membered enediyne model will be pursued (namely, dynemicin from Micromonospora chersina) selected for its unique architecture (an unprecedented fused enediyne-anthracycline), predominate biological activity, anticipated small gene locus size (excellent for production of dynemicin in 'genetically-friendly' heterologous hosts) and the opportunity for comparative genomics of the calicheamicin and dynemicin biosynthetic loci. The fundamental vision of this program remains constant - to present rational strategies from which to build a foundation of knowledge regarding 10-membered enediyne biosynthesis and self-resistance; the consequence of which will continue to provide pioneering discoveries in enzymatic transformation, tools for the rational biosynthetic modification of natural product drug leads, the potential for enediyne overproducing strains and possibly even an enediyne combinatorial biosynthesis program.

DESCRIPTION (provided by applicant): Aromatic polyketides differ from other polyketides by their characteristic polycyclic aromatic structures. These polyketides are widely distributed in bacteria, fungi and plants, and many of them are clinically valuable agents (e.g. tetracyclines, daunorubicin) or exhibit other fascinating biological activities. While the fundamental biosynthetic principles of aromatic polyketides has been extensively explored, little is known regarding the construction of many existing unique exocyclic carbon-carbon (C-C) attachments to these metabolites. The current proposal is designed to expand our understanding of this phenomenon by bringing together the study of two naturally occurring novel aromatic polyketide systems, which promise access to three distinct exocyclic aromatic polyketide C-C modifications. The first, hedamycin from Streptomyces griseoruber, offers one of the only naturally occurring di-C-glycosylation events as well as an additional type I PKS-directed C-alkyl chain substitution of the polycyclic ring system. The second, dynemicin from Micromonospora chersina, offers the only known naturally occurring fusion between an enediyne and aromatic polyketide. The intended studies are expected to lay a foundation to understand the unique mechanisms of these processes and also the potential to utilize these tools for combinatorial biosynthesis toward novel therapeutics. The first phase of this massive project is specifically intended to i) provide the necessary genetic tools and information to form reasonable mechanistic hypotheses for all three events, ii) to initiate in vivo and in vitro experiments explicitly designed to address the C-glycosylation and C-alkylation stages of hedamycin/pluramycin biosynthesis and Hi) attempt to combine aspects of all three aromatic C-C modification systems toward enhancing the diversification of aromatic polyketide structures. A particular focus of the final diversification strategies presented will be focused upon conjugation of these highly reactive antitumor antibiotics with known tumor-directing peptides.

While it is known that the sugar attachments of pharmaceutically important natural products often define their corresponding biological activity, until recently, efficient methods to alter these essential carbohydrate appendages, and thereby systematically explore their carbohydrate-based structure-activity relationships (SAR), was lacking. In recent years, we have developed two strategies to rapidly glycosylate natural products - in vitro glycorandomization (a chemoenzymatic process) and, more recently, neoglycorandomization (chemoselective-ligation based process). As Lab Program 2 of the University of Wisconsin National Cooperative Drug Discovery Group (UW NCDDG), this proposal outlines a systematic application of these two unique strategies toward diversification of a wide range of natural product parent scaffolds (including a number from UW NCDDG Program 1, Professor Shen) with previously indicated anticancer activities. Specifically, the proposed studies are designed to assess the general applicability of chemoenzymatic and chemoselective ligation-based glycorandomization to significantly enhance accessible natural product-based diversity and, in conjunction with the unique screening and downstream analysis capabilities of the UW NCDDG, will present perhaps the broadest correlation to date between the biological activity of natural products and the specific contributions invoked by their attached carbohydrates. More importantly, as mandated by the NCDDG, the parent scaffolds were also selected to significantly bias the ultimate outcome toward the identification of promising anticancer leads within the intended funding period and diversity directives will specifically evolve based upon the constant consultation of the novel screening/biological analysis/clinical components of the UW NCDDG

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Methyltransferases (MTs) are one of the tailoring enzymes involved in the biosynthesis of natural products. By engineering the MTs to transfer groups other than methyl will have the potential to generate chemotherapeutics. In this project, we wish to carry out binding studies with synthetic cofactors in order to make the enzyme promiscuous.

DESCRIPTION (provided by applicant): Natural products (NPs) and NP derivatives are an unrivaled, but highly under-represented, resource. Arguably among the most notable natural products discovered to date, the 10-membered enediynes - exemplified by the saccharide-fused calicheamicins (CLM) and the anthraquinone-fused dynemicins (DYN) - offer unprecedented molecular architecture, spectacular biological activity and demonstrated clinical utility. The objective of the first phase of this study (CA84374, years 1-4) was to i) clone and characterize the CLM gene cluster from M. echinospora, ii) develop the genetic tools to address CLM biosynthesis in M. echinospora, iii) investigate the mechanism(s) of CLM self-resistance, and iv) initiate aryltetrasaccharide biosynthetic studies. With these goals largely achieved and new tools/information in place, the second phase of this program (CA84374, years 5-9) was focused upon i) cloning and characterization of the DYN gene cluster from M. chersina, ii) development of genetic tools to address DYN biosynthesis in M. chersina, iii) initiating enediyne core biosynthetic studies, iv) structurally characterization of the CLM self-sacrifice resistance protein CalC, and v) the elucidation of key aryltetrasaccharide biosynthetic transformations (sugar N-oxidation, thiosugar formation and GT-catalyzed aryltetrasaccharide assembly). The successful completion of the majority of phase II aims enables the proposed course of study for this competitive renewal. Specifically, we will focus upon i) extending our understanding of enediyne core biosynthesis, ii) delineating the potential role of CalC in regulating CLM production in M. echinospora, iii) completing the study of key aryltetrasaccharide sugar nucleotide transformations (sulfur installation and sequential C-C alkylation of the N-alkyl dideoxypentose), iv) initiating an enediyne structural biology program and v) synthesizing and evaluating (neo)glycorandomized libraries of CLM and DYN. PUBLIC HEALTH RELEVANCE: This is a second competitive renewal of a productive program ((CA84374) targeting the biosynthesis of 10-membered enediynes (calicheamicin and dynemicin) - a novel class of anticancer natural products. The program is anticipated to provide pioneering discoveries in enzyme-catalyzed chemistries, new tools for the chemical diversification of complex natural products and unique anticancer lead compounds.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. To study the dynamics and ligand binding of glycosyltransferase, OleD.

? DESCRIPTION (provided by applicant): Although progress has been made in survival of patients with earlier stage colorectal cancer (CRC), only minimal improvement has been noted in patients with systemic metastases (Stage IV disease). Current chemotherapeutic agents, while highly effective at killing CRC cells, are limited by their systemic toxicity. If advances ar to be made in the survival of cancer patients, highly innovative strategies are required for more targeted delivery of anti-cancer agents directly to CRC metastases. Our ultimate translational goal is to develop a highly effective and less toxic approach to specifically deliver anti-cancer agents to CRC metastases. To achieve this goal, we have assembled a multidisciplinary and highly collaborative team who are at the forefront of molecular signaling pathways in CRC, CRC treatment modalities, medicinal chemistry, novel nanoparticle synthesis and delivery systems. Over the last two years, we have made significant progress to achieve our goals: (i) using a novel three-way junction (3WJ) motif, we have constructed thermodynamically and chemically stable three-branched RNA nanoparticles with an aptamer against receptors that can deliver a small molecule inhibitor or chemotherapeutic agent specifically to CRC metastases in the liver, (ii) we have constructed a variety of RNA nanoparticles using the pRNA-3WJ motif as a scaffold and have demonstrated in critical experiments that the resulting RNA constructs retained their folding and independent functionalities for specific cell binding, cell entry and cancer targeting, both in vitro and in vivo, (iii) we have shown that the RNA nanoparticles remain intact after systemic injection into mice and strongly bind to tumors with little accumulation in normal organs or tissues; these RNA constructs are non- toxic, non-immunogenic, and display favorable pharmacological profiles in mice, and (iv) we have demonstrated localized in vivo delivery of pRNA-3WJ to CRC xenografts and liver metastases. Thus, the central hypothesis of our proposal is that CRC receptor-specific delivery of chemotherapeutic agents using our pRNA-3WJ nanoparticles will provide a safe, effective strategy to selectively target and inhibit CRC metastasis. To address our hypothesis, we have designed experiments with the following Specific Aims: 1) to construct pRNA-3WJ nanoparticles coupled with anti-cancer agents and analyze their stability, cellular uptake and anti-proliferative effects in vitro; 2) to determine te pharmacokinetics, stability, safety and drug delivery of pRNA-drug conjugates in vivo; and 3) to evaluate the selective delivery and in vivo anti-tumor effect of pRNA- 3WJ nanoparticles coupled with anti-cancer agents. In summary, our enthusiasm for our current proposal is driven not only by its inherent scientific importance, but also by its translational potential, clinical impact, and the possibility to provide a more effective and less toxic delivery system targeting CRC metastases.

? DESCRIPTION (provided by applicant): This Multi-PD/PI application seeks to develop resources that will better enable studies of a highly regenerative amphibian, the Mexican axolotl (Ambystoma mexicanum). There is need to probe axolotl regeneration more deeply, with the same powerful approaches that have proven so effective in genetic model organisms. Specific Aim 1 will accomplish the first chemical genetic screen of axolotl regeneration using an embryo tail regeneration assay. Using pre-feeding axolotl embryos that are efficiently reared in micro-titer plates, up to 10,000 soluble chemicals from commercial, clinical-stage, and novel natural products libraries will be tested for impact on tail regeneration. Preliminary data show that the chemical screen and tail regeneration assay are likely to identify new molecules that impact regeneration. Positive hits from this screen, including previously identified inhibitors of Wnt, Tgf?, and Fgf signaling, will be reported to the community and investigated further under Specific Aim 2, using cellular and transcriptional approaches. In particular, assays will be developed to assess chemical effects on formation of the wound epidermis, which acts as an early signaling center in the recruitment of progenitor/stem cells. Also, assays will be developed to assess cellular proliferation and de- differentiation, two processes that are associated with endogenous regeneration. Genes found to be expressed differently between control and chemically treated embryos will be prioritized for knock out using the CRISPR/Cas9 method. For each gene target, two gRNA pairs will be designed and injected into single cell axolotl embryos. Embryos will be reared to assess viability, and then administered tail amputations to confirm CRISPR gene editing and test for regeneration competence. Embryos associated with CRISPRs that block or cause abnormal regeneration, will be prioritized for founding stable lines. Specific Aim 3 will generate fluorescent reporter lines to assay signaling activity through major pathways known to function in regeneration and will also compare efficacies of two methods for developing gene-specific reporter lines, using as tester loci genes already known to mark cell populations critical for regenerative responses (neural stem cells, glia, muscle satellite cells). The chemical and genetic hits, and biological information arising from this model will be shared through a community website (Sal-Site). The proposed transgenics will be distributed by the Ambystoma Genetic Stock Center. Overall, this project integrates expertise across chemical screening, pharmacology, histology, transcription, transgenesis, and vertebrate biology to discover reagents and develop tools that are needed to enhance the axolotl for stem cell and regenerative biology.

? DESCRIPTION (provided by applicant): Metastatic colorectal cancer (CRC) is difficult to treat and patients have few long term effective therapeutic options. The aggressiveness of this disease is in part driven by the aberrant expression of oncoproteins. At the molecular level, cap-dependent translation of the precursor oncogenic mRNAs is frequently activated. Specifically this occurs via 4E-BP1 phosphorylation which, when not phosphorylated, functions as a mRNA translation repressor downstream from mTOR. We recently discovered that activated signaling via the PI3K/AKT and RAS/RAF/MEK/ERK pathways cooperate to promote CRC progression by convergent phosphorylation of 4E-BP1. Our work further demonstrated that 4E-BP1 phosphorylation-mediated oncogene translation functions as a critical node that integrates oncogenic signals of the AKT and ERK pathways for CRC tumorigenesis and metastasis. Moreover, we found that CRC resistance to upstream kinase targeted therapy is associated with incomplete inhibition of 4E-BP1 phosphorylation. Notably, genetic blockade of cap-dependent translation by a dominant active and non-phosphorylated 4E-BP1 mutant can effectively suppress tumor growth and metastasis in the mouse models of CRC. Our overarching hypothesis is that directly targeting 4E-BP1 phosphorylation- mediated oncogene translation represents a novel strategy for cancer drug development and therapy. Using a cap-dependent translation-based reporter assay, we recently identified naturally occurring pyranonaphthoquinones that act as selective inhibitors of 4E-BP1 phosphorylation in a manner that is mechanistically distinct to existing mTOR inhibitors. The primary goals of the proposed studies are to determine the fundamental mechanism of pyranonaphthoquinone-based inhibition of 4E-BP1 phosphorylation and identify optimized analogs with suitable in vitro and in vivo potency and selectivity. Cumulatively, the proposed studies offer high potential for the identification and development of structurally and functionally novel agents to target the translational control of CRC progression and metastasis with the potential to define new molecular probes and early stage leads for first-in-class targeted therapies to treat CRC.

? DESCRIPTION (provided by applicant): Methyltransferase (MT)-catalyzed S-adenosyl-L-methionine (AdoMet)-dependent methylation is essential to all walks of life and alterations in methylation-dependent processes have direct relevance to microbial/fungal/viral pathogenesis and human disease. Yet, for many MTs, there is a lack of correlative fundamental knowledge regarding specific MT function and corresponding impact upon cellular fate and/or pathogenesis/disease. In addition, while simple chemical or MT-catalyzed methylation of a drug/drug lead can dramatically impact its corresponding ADMET (absorption, distribution, metabolism, excretion and/or toxicity), the structural complexity of many natural products often prohibits doing so in the context of natural products-based drugs/lead development. This proposal seeks to develop general chemoenzymatic alkylation strategies and reagents for that are expected to broadly facilitate the fundamental study, annotation and application of MTs. A centerpiece to the proposed universal platform development is the study, engineering and application of permissive methionine adenosyltransferases (MATs) and MTs, where the model MTs selected represent broad catalytic diversity (C-methylation, O-methylation and N-methylation) and directly act upon a selected set of complex natural product-based drugs, validated clinical candidates or marketed agricultural products. The proposed studies will integrate the chemical synthesis and application of unique methionine (Met) analogs, MAT/MT structure determination, high throughput MAT/MT assay development/application, structure-guided MAT/MT directed evolution, microbial strain engineering, complex natural product (NP) structure elucidation and bioactivity assessment for NP analogs generated. The anticipated outcomes of this study include highly permissive/proficient MATs/MTs engineered for medicinal chemistry applications, novel functional AdoMet orthologs designed as alternative alkyl donors and/or with improved stability, an expanded understanding of MAT/MT structure-activity relationships of potential relevance to MAT/MT inhibitor design, single vessel chemoenzymatic strategies to enable complex NP differential alkylation, engineered microbial strains to enable complex NP differential alkylation and functional MT annotation, and unprecedented differentially-alkylated NP analogs with potential therapeutic and/or agricultural applications. Within this context, the proposed studies will also provide the first bioorthogonal strategy to functionally annotate, interrogate or exploit a single MT within a cell containing a full complement of competing native MTs. While NP methylation has been selected as the model for platform development, it is important to note that reagents and concepts developed will likewise enable the similar study, annotation and application of other class I MTs relevant to cellular development, human disease or microbial/fungal/viral pathogenesis.

Tuberculosis (TB)?which is primarily caused by the bacterial pathogen Mycobacterium tuberculosis (Mtb)?is an ancient disease that remains one of the deadliest communicable diseases worldwide. A paramount concern heading into the future is the rapid rise in drug-resistant TB. The World Health Organization estimated 480,000 cases of TB with 190,000 deaths in 2014 with resistance to the first-line anti-TB drugs isoniazid and rifampicin. Furthermore, totally drug-resistant Mtb has now been documented in multiple countries including the United States. The capuramycin family of glycosylated nucleoside antibiotics are excellent candidates for anti-TB drug discovery and development because they (i) are considered new chemical entities with several unusual structural features compared to all antibiotics including clinical anti-TB drugs, (ii) target a novel and essential enzyme (translocase 1; TL1) in cell wall biosynthesis, (ii) have exceptional anti-Mtb activity in vitro and in vivo, (iv) are bactericidal and kill Mtb faster than any first-line anti-TB drug in vitro, and (v) have no toxicity. Our primary objectives in this proposal are to define a biosynthetic mechanism for the assembly of the unusual unsaturated hexuronic acid component in capuramycins (Aim 1) and establish complementary chemical (via neoglycorandomization; Aim 2) and biosynthetic (via native and nonnative glycosyltransferases; Aim 3) platforms for rapidly generating novel hexuronic acid-substituted capuramycins that can be screened for TL1 inhibition, anti-Mtb activity, and improved pharmacological properties. Additionally, these novel capuramycin analogues will be screened as potential substrates or inhibitors of the phosphotransferase CapP, which covalently modifies capuramycin as a strategy of self-resistance within the producing strain and is potentially a widespread resistance mechanism. It is expected that, upon completion of the aims, a new biosynthetic mechanism for sugar incorporation and modification will be defined. Furthermore, the completion of the aims will provide the first practical, comprehensive strategy to rapidly interrogate/modulate the fundamental features of capuramycin core pharmacophore, which will not only be important for the clinical development of capuramycin but can be applied to other glycosylated nucleoside antibiotics, of which dozens are now known with diverse biological activities.

ABSTRACT The 10-membered enediynes [exemplified by calicheamicin (CLM), esperamicin (ESP) and dynemicins (DYN)] are arguably among the most renowned natural products (NPs) discovered to date by virtue of their unprecedented complex molecular architectures, notable anticancer and anti-infective potencies and, in the case of CLM, demonstrated clinical utility. The current study builds on a longstanding collaborative effort of achievement and discovery relating to key aspects of 10-membered enediyne biosynthesis as well as parallel innovative efforts to co-opt key biosynthetic catalysts for synthetic applications. The studies put forth will take advantage of this strong foundation and a powerful combination of genetic, biochemical, chemical and protein structural tools to elucidate the remaining unusual biosynthetic transformations and to exploit select catalysts for enediyne non-native modification. Specifically, aims 1 and 2 will focus on extending our understanding of the fundamental steps of enediyne core biosynthesis common to CLM/DYN/ESP, DYN anthraquinone biosynthesis and a selected set of unique tailoring reactions (CLM/ESP thiosugar sulfur installation and aminopentose N- alkylation, ESP C6-hydroxylation and O-glycosylation). In parallel, aim 3 will focus on tactical structural studies to augment both aims 1 and 2 and the structural study of ?unknowns? to facilitate functional annotation. Additional studies in aim 2 with key catalysts and corresponding non-native substrates are designed to assess the potential for strategic installation of chemoselective handles to enable novel approaches for facile, mild bioconjugation of CLM to tumor-targeting mAbs (in collaboration with Pfizer).

PROJECT SUMMARY/ABSTRACT The Drug Discovery, Delivery and Translational Therapeutics (DT) Program at the Markey Cancer Center (MCC) is scientifically focused on identifying novel targets and biomarkers and discovering and developing new drugs targeting these biomarkers. The MCC catchment area population has both a high cancer risk related to excessive carcinogen exposure and lack of access to cutting-edge clinical trials due to geographical isolation and poor socioeconomic status. The DT program vision is to understand the unique molecular and phenotypic markers of cancer in Kentucky as well as barriers to accessing care and integrate that knowledge to inform drug discovery, development, and delivery of early phase clinical trial efforts for a hard-to-reach Appalachian Kentucky population. MCC investigators are international leaders in biomarker discovery (Theme 1) with ongoing translational studies including more than 600 participants, evaluating the role of environmental carcinogens and identifying biomarkers of lung cancer. DT pharmaceutical scientists work to discover and develop new anticancer agents targeting identified mutations and phenotypes (Theme 2), partnering with Cancer Cell Biology and Signaling (CS) and Genomic Instability, Epigenetics, and Metabolism (GEM) program members. For example, a novel modulator of 4E-BP1 phosphorylation, a validated colon cancer target, was identified from the Appalachian natural products repository. DT investigators lead clinical trials focusing on cancers relevant to the catchment area (Theme 3) and have enrolled more than 500 patients to lung, colon and ovarian interventional treatment and diagnostic trials. They regularly partner with Cancer Prevention and Control (CP), CS and GEM program members to inform and advance MCC basic science, for example, translating early identification of the anticancer activity of PAR-4 in CS to clinical trials focused on a PAR-4 secratagogue. DT is a cross-disciplinary program of 47 investigators from 6 colleges and 18 departments who work together to develop novel anticancer therapies and translate these therapies into the clinic. This productive program has total annual external cancer-related funding of $8.5M ($5.9M annual direct costs, of which 28% is from the NCI). Members published 366 publications over the current funding period, 99 (27%) of which are inter-programmatic, 84 (23%) are intra-programmatic, and 189 (52%) are inter-institutional. The DT program Co-leaders, Drs. Jill Kolesar and Jon Thorson, have a long-standing collaboration and bring complementary expertise in biomarker discovery, drug development, and early clinical trials. Both direct key resources supporting the DT program, the MCC Precision Medicine Center and the UK Center for Pharmaceutical Innovation, respectively. Each leader brings critical strengths including local, national and international collaborations, entrepreneurial relationships, and active participation in NCI initiatives. Taken together, the DT program has a cohesive and collaborative team that translates novel biomarkers into targets for effective anticancer treatments.

PROJECT SUMMARY/ABSTRACT ? ADMINISTRATIVE CORE We propose to establish a Center of Biomedical Research Excellence (COBRE) in Pharmaceutical Research and Innovation at the University of Kentucky (UK). CPRI will serve as a comprehensive multidisciplinary center focused on translational chemical biology [the nexus of chemical biology (the application of chemical biology principles to develop validated probe/models to advance our understanding of biology) and pharmaceutical science (the application of pharmaceutical principles to advance leads/materials/devices that address unmet clinical needs)]. The COBRE will leverage and develop unique translational chemical biology research support infrastructure/expertise to facilitate junior faculty mentorship and career development, innovative biomedical research probe/tool/model/materials development and validation, and the early advancement of potential ?translatable? assets. Within this context, the CPRI Administrative Core will support center communication, coordination, oversight and integration of all COBRE operations, activities, projects, pilots and research support cores. The Core will provide fiscal management and resource oversight, overall coordination of mentoring and career development activities, resource allocation, and program evaluation to ensure program effectiveness. The Administrative Core will also facilitate interactions with related UK programs and centers to foster synergy and coordination of resources. The Core will facilitate the Center?s engagement of junior faculty, researchers, and mentors from UK Colleges (Agriculture, Food & Environment; Arts and Sciences; Engineering; Medicine; Pharmacy and others) to create a vibrant center that seamlessly integrates resources, personnel and knowledge across disciplinary boundaries, and fosters scientific innovation and career development.

PROJECT SUMMARY/ABSTRACT - OVERALL We propose to establish a Center of Biomedical Research Excellence (COBRE) in Pharmaceutical Research and Innovation at the University of Kentucky (UK). CPRI will serve as a comprehensive multidisciplinary center focused on translational chemical biology [the nexus of chemical biology (the application of chemical biology principles to develop validated probe/models to advance our understanding of biology) and pharmaceutical science (the application of pharmaceutical principles to advance leads/materials/devices that address unmet clinical needs)]. The COBRE will leverage and develop unique translational chemical biology research support infrastructure/expertise to facilitate junior faculty mentorship and career development, innovative biomedical research probe/tool/model/materials development and validation, and the early advancement of potential ?translatable? assets. Key COBRE infrastructure to be developed and implemented in Phase I includes the CPRI Administrative Core and two research support cores (the Translational Core and the Computational Core) and the COBRE will also further develop and leverage the COBRE for Molecular Medicine?s Organic Synthesis Core to support CPRI junior faculty projects/pilots. CPRI will initially support four outstanding junior investigators working in three therapeutic areas (cancer, infectious disease and cardiovascular disease). Our overarching hypothesis is that CPRI?s translational chemical biology focus presents a distinctly unique UK platform to: i) engage, integrate and mentor junior faculty from a broad range of fundamental, applied and clinical disciplines; ii) facilitate the development of new impactful probes/tools/models and advance innovative transdisciplinary research; and iii) bridge the gap between basic academic research discoveries and commercial/clinical application via education, mentorship and key support infrastructure. CPRI?s distinct focus on translational chemical biology and early translation fills a notable preclinical research gap for many of UK?s exceptional research centers strategically focused on understanding, treating and preventing the major diseases that contribute to Kentucky?s disproportionate health challenges. Anticipated COBRE outcomes include an increase in the number and diversity of UK junior faculty engaged in translational chemical biology research and better-prepared to develop and employ innovative biomedical probes/tools/models/materials, new sustainable UK translational chemical biology research support infrastructure and capabilities, an increase in the number of UK junior faculty with independent research funding, and a boost in the number potentially ?translatable? UK preclinical assets.