Identification and characterization of a novel glutaminase inhibitor.

In humans, there are two forms of glutaminase (GLS), designated GLS1 and GLS2. These enzymes catalyse the conversion of glutamine to glutamate. GLS1 exists as two isozymes: kidney glutaminase (KGA) and glutaminase C (GAC). Several GLS inhibitors have been identified, of which DON (6-diazo-5-oxonorleucine), BPTES (bis-2-(5-phenylacetamido-1, 3, 4-thiadiazol-2-yl) ethyl sulphide), 968 (5-(3-Bromo-4-(dimethylamino)phenyl)-2,2-dimethyl-2,3,5,6-tetrahydrobenzo[a]phenanthridin-4(1H)-one) and CB839 (Telaglenastat) are the most widely used. However, these inhibitors have variable efficacy, specificity and bioavailability in research and clinical settings, implying the need for novel and improved GLS inhibitors. Based on this need, a diverse library of 28,000 compounds from Enamine was screened for inhibition of recombinant, purified GAC. From this library, one inhibitor designated compound 19 (C19) was identified with kinetic features revealing allosteric inhibition of GAC in the µm range. Moreover, C19 inhibits anti-CD3/CD28-induced CD4+ T-cell proliferation and cytokine production with similar or greater potency as compared to BPTES. Taken together, our data suggest that C19 has the potential to modulate GLS1 activity and alter metabolic activity of T cells.

An in vitro Study of Protein Adsorption to Biocompatible Coatings.

The motivation for these experiments was to investigate the amount and type of protein adsorption on surfaces that can be used as protective coatings on membrane based in vivo devices. Adsorption of proteins to a selection of biocompatible coatings (titanium oxide, diamond-like carbon, parylene C) and typical construction materials for Micro Electro Mechanical Systems (silicon, silicon nitride), were investigated during in vitro tests. The samples were incubated in human liver extract and bovine serum albumin (BSA) for up to 12 hours. The amount of protein adsorption was found to be low for all surfaces. Measurements of bound Iodine-125 labeled BSA, showed a protein adsorption of up to 0.2 µg BSA/cm2. The specific proteins adsorbed to the surfaces after incubation in human liver extract were identified using mass spectrometry. Most of the identified adsorbed proteins were intracellular, but plasma proteins like Immunoglobulin (Ig) and serum albumin as well as hemoglobin were also identified.