Metabolic reprogramming in tumors represents a potential therapeutic target. Herein we used shRNA depletion and a novel lactate dehydrogenase (LDHA) inhibitor, GNE-140, to probe the role of LDHA in tumor growth in vitro and in vivo. In MIA PaCa-2 human pancreatic cells, LDHA inhibition rapidly affected global metabolism, although cell death only occurred after 2 d of continuous LDHA inhibition. Pancreatic cell lines that utilize oxidative phosphorylation (OXPHOS) rather than glycolysis were inherently resistant to GNE-140, but could be resensitized to GNE-140 with the OXPHOS inhibitor phenformin. Acquired resistance to GNE-140 was driven by activation of the AMPK-mTOR-S6K signaling pathway, which led to increased OXPHOS, and inhibitors targeting this pathway could prevent resistance. Thus, combining an LDHA inhibitor with compounds targeting the mitochondrial or AMPK-S6K signaling axis may not only broaden the clinical utility of LDHA inhibitors beyond glycolytically dependent tumors but also reduce the emergence of resistance to LDHA inhibition.

Metabolic activation of a drug leading to reactive metabolite(s) that can covalently modify proteins is considered an initial step that may lead to drug-induced organ toxicities. Characterization of reactive metabolites is critical to designing new drug candidates with an improved toxicological profile. High performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) predominates over all analytical tools used for screening and characterization of reactive metabolites. In this review, a brief description of experimental approaches employed for assessing reactive metabolites is followed by a discussion on the reactivity of acyl glucuronides and acyl coenzyme A thioesters. Techniques for high-throughput screening and quantitation of reactive metabolite formation are also described, along with proteomic approaches used to identify protein targets and modification sites by reactive metabolites. Strategies for dealing with reactive metabolites are reviewed. In conclusion, we discuss the challenges and future needs in this field of research.

Alkanethiol-protected silver clusters of average diameter 4.0 ( 0.5 nm form single-phase superlattice solids, and their X-ray powder diffractograms have been fully indexed to single cubic unit cells. Whereas alkanethiols with five or more carbon atoms form superlattices, the corresponding cluster with four carbons yield only separated clusters. The superlattice solids can be recrystallized from nonpolar solvents. No such superlattices are seen for the corresponding gold clusters. The superlattice collapses upon heating, but the solid retains the structure even at 398 K, much above the melting point of crystalline alkanes and the corresponding self-assembled monolayer. In situ variable-temperature X-ray diffraction investigations did not show any solid-state phase transitions in the superlattice. Temperature-dependent infrared spectroscopy reveals the melting of the alkyl chain, and it is seen that the chain as a whole achieves rotational freedom prior to the collapse of the superlattice. Calorimetric investigations show distinct monolayer and superlattice melting transitions. The chemical nature of the cluster-molecule interaction is similar to that of the previously investigated gold and silver systems, as revealed by NMR, mass, infrared, and X-ray photoelectron spectroscopies and thermogravimetry analyses. Conductivity measurements clearly manifest the superlattice melting transition. Diffusion constants in solution measured by NMR show that the relative decrease in the diffusion constant with increasing monolayer chain length is smaller for silver than for gold, suggested to be a signature of intercluster interaction even in solution. Corroborative evidence is provided by the variable-temperature UV/vis investigations of the clusters.