Disease relevance of HTS

• The unique, high-affinity binding of cyanovirin-N (CV-N), a potent anti-human immunodeficiency virus (HIV) protein, to the HIV envelope glycoprotein gp120, was exploited to develop an HTS assay in an attempt to discover small-molecule mimetics of CV-N [1].
• After physical training, in HTS PI variance increased suggesting a decrease in frequency modulated sympathetic activity and increase in vagal modulation of heart rate in mild hypertension [2].
• Conclusion: In this model of rat with severe hemorrhagic shock, small volume resuscitation with HTS is more effective than NS in reducing immunologic disorders and promoting a more balanced profile of T-lymphocyte subpopulations regulating network [3].
• A 3-year-old boy with Wilms' tumor, post operative left nephrectomy stage, had HTS on day 99 of the combined chemotherapy which lasted for more than 20 days [4].
• The aim is to use the NRU assay as one test component of HTS strategies for both acute oral toxicity and acute skin irritation, enabling the rejection of the most toxic materials and prioritisation of other materials for further testing [5].

High impact information on HTS

• Structure-based screening as applied to human FABP4: a highly efficient alternative to HTS for hit generation [6].
• A Chemical Genetics Approach for the Discovery of Apoptosis Inducers: From Phenotypic Cell Based HTS Assay and Structure-Activity Relationship Studies, to Identification of Potential Anticancer Agents and Molecular Targets [7].
• The discovery of non-peptidic low molecular weight compounds able to inhibit PTP1B, by means of docking procedures and HTS screening, and the presence of secondary binding sites on PTP1B afforded new potent and selective inhibitors; several leads devoid of negative charges were also found [8].
• We have used whole-cell and perforated patches to study ionic currents induced by hypotonic extracellular solutions (HTS, 185 mOsm instead of 290 mOsm) in endothelial cells from human umbilical veins [9].
• The HTS-induced currents may therefore be modulated by cleavage products of PLA2, but not by messengers downstream of arachidonic acid [9].

Biological context of HTS

• The identification of N-phenyl nicotinamides as a novel series of inducers of apoptosis demonstrates that our cell- and caspase-based HTS assay is useful for the discovery and optimization of potentially novel anticancer agents [10].
• It is now possible, comprehensively and systematically, to enumerate, clone, produce and screen all secreted proteins, by building upon knowledge accumulated over the past two decades in HTS, genomics and parallel protein expression technologies [11].
• In order to identify ideal lipofection reagents in a HTS, we have developed an automated lipofection method for the transfer of reporter genes into cells and for determination of the lipofection results [12].
• The HTS method facilitates characterization of the transfection properties (efficiency and cytotoxicity) of large numbers of (cationic) lipids in various adherent cell types [12].
• Although similar cell fusion systems have been described previously, they have not met the requirements for HTS due to complexity, throughput and reagent cost [13].

Anatomical context of HTS

• In this perspective we address a related, but very focused, issue that is increasingly important for many of us in the HTS community: the use of stably transfected cell lines as an integral part of screening strategies [14].
• Caco-2 monolayers obtained by 3-day culture in the BIOCOAT HTS Caco-2 Assay System, developed by Becton Dickinson Bioscience, showed much higher permeability to hydrophilic compounds, such as mannitol, compared with those obtained by the standard 21-day culture system, due to the leaky structure of cell junctions [15].
• Small volume resuscitation with HTS also induced peripheral CD8(+) lymphocytes to a certain extent, whereas NS resuscitation showed no effect in this respect [3].

Associations of HTS with chemical compounds

• Strategies for optimizing bioassays include: considering solubility in HTS-library design; early screening for solubility; improving storage and handling of DMSO stocks; optimizing dilution protocols; and ensuring that low-solubility compounds are fully solubilized in bioassays [16].
• Recent literature has highlighted the importance of fundamental physicochemical properties in HTS and ADME-Tox: high lipophilicity, low dimethyl sulfoxide solubility, low aqueous solubility, aggregation and nonspecific binding to proteins and phospholipids [17].
• We constructed an HTS assay based on the interaction of europium-labeled CV-N with recombinant glycosylated gp41 ectodomain to support identification of small-molecule mimetics of CV-N that might be developed as antiviral drug leads [18].
• Assay parameters were validated using A431 cell extracts containing EGF Receptor. The autophosphorylation capture assay is a simple and rapid method which can be adapted for use with robotics for qualitative HTS of potential inhibitors to any tyrosine kinase of interest [19].
• Microbial HTS has been implemented at Rhône-Poulenc Rorer through the development of a dedicated robotic platform [20].

Physical interactions of HTS

• To identify novel Hsp90 inhibitors, a highly robust time-resolved fluorescence resonance energy transfer (TR-FRET)-based HTS assay that measures the binding of biotinylated geldanamycin (biotin-GM) to the His-tagged human Hsp90 N-terminal ATP-binding domain (Hsp90N) was developed [21].

Other interactions of HTS

• We describe an HTS-compatible method using the peptide substrate PLGLAARK, labeled at N- and C-termini with CyDye fluors Cy3 and Cy5Q, respectively, which is cleavable by MMP-1, -2, -3, -7, -9, and -13 [22].
• In the other test (Lactobacillus casei, thymidylate synthase (TS), human HTS) no or poor activity was detected in both series of compounds [23].
• The FP HTS system provides a specific, sensitive and reproducible methodology for studying and screening NPFF receptor ligands [24].
• The data indicate that HTE and HTS function as positive and negative regulatory elements that control human TIMP-1 expression [25].
• A TR-FRET assay has been developed and evaluated for HTS for TRAF6 inhibitors [26].

Analytical, diagnostic and therapeutic context of HTS

• Small-molecule microarrays, protein arrays and cell-based arrays and conventional DNA arrays as well as microfluidic approaches in HTS are discussed in this review [27].
• High-throughput bioassay-guided fractionation: a technique for rapidly assigning observed activity to individual components of combinatorial libraries, screened in HTS bioassays [28].
• The widespread use of HTS and combinatorial chemistry techniques has led to the generation of large amounts of pharmacological data, which, in turn, has catalyzed the development of computational methods designed to reduce the time and cost in identifying molecules suitable for pharmaceutical development [29].
• The rats were randomly divided into Sham group, HTS group (hypertonic saline resuscitation group) and NS group (normal saline resuscitation group) [3].
• A diverse library of druggable, purified and quantitated molecules was assembled and standardized enzymatic assays were performed in a microfluidic format that provided very accurate and precise inhibition data allowing for development of SAR directly from the primary HTS [30].

References