Administrative | Protein Analytical | Imaging | Organic | Proteomics | Viral Production

Introduction and Purpose

The Organic Synthesis Core provides "small molecule" probes to investigators as components of their proposed projects. An important component of this core is consultation in which the staff of the core understand the purpose behind the request, explain options regarding the requested synthesis (i.e., time commitment, chemical stability, proper handling, material costs) and discuss alternatives. The core provides unique probes for addressing biochemical problems and participates in the writing of grants in support of projects using these probes.

Specific Functions of Core

The core furnishes modified drugs and natural products tailored to include suitable functional groups for linkage to proteins, peptide-nucleic-acid (PNA) mimics, and polymers or with suitable markers for the detection and monitoring of a particular drug or natural product. Tailoring include the incorporation of mass-spectrometry markers, fluorescent tags, photoactive groups, or radiolabels, as needed. The laboratory maintains equipment needed for organic synthesis and utilizes University infrared, ultraviolet, mass spectrometry and nuclear magnetic resonance instrumentation to characterize adducts and establish their purity. External laboratories are used to perform combustion analyses. Training opportunities are available for graduate students, technicians and postdoctoral fellows from other research groups who wish to learn a particular technique and subsequently import it into their research group. The core provides access to equipment (e.g., melting point device, low-pressure distillation equipment, specialized glassware such as Dean-Stark traps, Soxlet extractors, etc.) and chromatographic systems not generally available in biochemical laboratories and provides access to this equipment to any investigators who need it.

Utilization in Individual Projects

The Core Director has met with each of the COBRE investigators to assess their needs and determine future directions where the core might be of assistance. In support of Dr. Christian Paumi's work on the multidrug resistance protein (MRP) transporters, which transport substrates in the form of glutathione derivates for cellular and biochemical assays, the core will synthesize various modified glutathione derivatives. One such probe requested by Dr. Paumi, S-(2,4-dinitrophenyl)glutathione (DNP-SG) was recently completed and discussions regarding fluorescent analogs are proceeding. In support of Dr. Francesc Marti's work on peripherally induced regulatory T cells (iTreg) the core will synthesize eicosanoid derivatives for cellular and biochemical analyses related to the regulation of transcriptional activity of peroxisome proliferator-activated receptors (PPARg). The director has had considerable experience with eicosanoid synthesis and the developing of photoactive eicosanoids. We are also, in collaboration with Dr. Andrew Morris, developing an analytical scheme for detecting and measuring various fatty acids and anticipate that this analytical tool may be useful in the measurement of polyunsaturated fatty acids (PUFAs) that interfere with the generation of iTreg. In support of Dr. Craig Vander Kooi's work on neuropilin and its vascular endothelial growth factor (VEGF) ligands, the core will synthesize glycan analogs. In addition the core will continue to synthesize compounds for past COBRE PIs, pilot PIs, and other faculty.

Broad Impact

In addition to consultation with the COBRE investigators, the core maintains an open-door policy for consultation and experimental work with groups across the University. The core is involved in numerous ongoing projects with junior and senior investigators. The following is a representative list of the projects and their progress completed during the past twelve months:

1. preparation of a phenothiazine-linked Sephadex polymer for the purification of calmodulin (completed project, material is in active use);

2. the use of pyridinium salts as "mass spectrometric tags" that produce strong parent ions in the analysis of fatty acids (progress report: syntheses complete; polymer-bound reagents now in preparation; analytical work with current agents is promising);

3. the synthesis of cantharidin-like inhibitors of protein phosphatase 1 and their linkage to PNA oligonucleotides for studies of pre-RNA splicing. To date 40+ analogs have been prepared; active compounds submitted to NCI for evaluation; one compound linked to a morpholino oligo undergoing analysis; three grant applications submitted;

4. the synthesis of "dimeric" estrogens optimized for their ability to stimulate estrogen-mediated gene expression events selectively target. To date 10+ compounds synthesized of varying hydropathy; active compounds under investigation; R21 submitted;

5. the synthesis of a neprilysin inhibitor. Synthesis completed, undergoing evaluation;

6. synthesis of a ceramidase inhibitor/probe. Synthesis of model system completed, work on target soon to begin;

7. synthesis of a germacrene natural product in collaboration with College of Agriculture investigator. Preliminary work completed);

8. synthesis of a biotinylated levetriacetam, an anti-epileptic drug. Synthesis completed; and

9. synthesis of a stable analog of a naturally occurring lipid that appears in brain injury studies. Work now underway.