BET proteins: Investigating BRDT as a potential target for male contraception.

While many contraception options are available for women, birth control methods for men are limited to condoms and vasectomy. Past research into male contraceptives has focused on hormonal options but the associated side effects have thus far precluded this method from reaching the market. Non-hormonal male contraceptives and vas occlusion have also been explored, but to date no method has progressed past clinical testing. Recent interest in epigenetic research has unveiled a new potential non-hormonal male contraceptive target: the testis-specific bromodomain BRDT. Potent inhibitors for bromodomain-containing proteins are described in the literature, but a BRDT-specific compound has yet to be designed, prepared and tested. The high similarity between bromodomain proteins of the BET family makes development of selective and specific inhibitors both difficult and necessary. Selective inhibition of BRDT by a small molecule is an exciting new target in the search for a new non-hormonal male contraceptive.

Reducing the Impact of Perfusion Medical Waste on the Environment.

The U.S. healthcare system generates more than five billion pounds of waste each year. Waste disposal has become a serious environmental problem facing healthcare institutions. The operating room is the second largest source of hospital waste, and no current standards exist regarding perfusion waste reuse or recycling. A typical perfusion circuit produces approximately 15 pounds of plastic that ends up incinerated once used. Contaminated perfusion circuits consisting primarily of polyvinyl chloride (PVC) and polycarbonate are difficult to sterilize, reuse, or recycle. A literature review of Internet-based and peer-reviewed publications was conducted to identify all resources that describe sterilizing, dechlorinating, reusing, and recycling of medical-grade disposable products. There are several chemical methods available to re-harvest PVC after it has been properly decontaminated and melted down. Dichlorination by near-critical methanol shows promise in the recovery of additives such as plasticizers, stabilizers, and lubricants. The reinjection of PVC may have ecological and economic advantages. Dechlorinated PVC also creates a less toxic by-product when incinerated. Although this process is not recycling, it lessens the impact of poisonous chlorine gas release into the atmosphere. Sterilizing, dechlorinating, and recycling the perfusion circuit may be a promising avenue for reducing the ecological impact of perfusion waste. Although an economically sensitive mode of reusing, reducing, and recycling a circuit does not currently exist, this presentation will explore the perfusion waste dilemma and present potential solutions in hopes of promoting future reuse and recycling opportunities.

BET Bromodomain Inhibitors with One-Step Synthesis Discovered from Virtual Screen.

Chemical inhibition of epigenetic regulatory proteins BrdT and Brd4 is emerging as a promising therapeutic strategy in contraception, cancer, and heart disease. We report an easily synthesized dihydropyridopyrimidine pan-BET inhibitor scaffold, which was uncovered via a virtual screen followed by testing in a fluorescence anisotropy assay. Dihydropyridopyimidine 3 was subjected to further characterization and is highly selective for the BET family of bromodomains. Structure-activity relationship data and ligand deconstruction highlight the importance of the substitution of the uracil moiety for potency and selectivity. Compound 3 was also cocrystallized with Brd4 for determining the ligand binding pose and rationalizing subsequent structure-activity data. An additional series of dihydropyridopyrimidines was synthesized to exploit the proximity of a channel near the ZA loop of Brd4, leading to compounds with submicromolar affinity and cellular target engagement. Given these findings, novel and easily synthesized inhibitors are being introduced to the growing field of bromodomain inhibitor development.

Protein-observed (19)F-NMR for fragment screening, affinity quantification and druggability assessment.

NMR spectroscopy can be used to quantify the binding affinity between proteins and low-complexity molecules, termed 'fragments'; this versatile screening approach allows researchers to assess the druggability of new protein targets. Protein-observed (19)F-NMR (PrOF NMR) using (19)F-labeled amino acids generates relatively simple spectra that are able to provide dynamic structural information toward understanding protein folding and function. Changes in these spectra upon the addition of fragment molecules can be observed and quantified. This protocol describes the sequence-selective labeling of three proteins (the first bromodomains of Brd4 and BrdT, and the KIX domain of the CREB-binding protein) using commercially available fluorinated aromatic amino acids and fluorinated precursors as example applications of the method developed by our research group. Fragment-screening approaches are discussed, as well as Kd determination, ligand-efficiency calculations and druggability assessment, i.e., the ability to target these proteins using small-molecule ligands. Experiment times on the order of a few minutes and the simplicity of the NMR spectra obtained make this approach well-suited to the investigation of small- to medium-sized proteins, as well as the screening of multiple proteins in the same experiment.