scholarly journals Identification of phosphorylase kinase as a novel therapeutic target through high-throughput screening for anti-angiogenesis compounds in zebrafish

Oncogene ◽  
2011 ◽  
Vol 31 (39) ◽  
pp. 4333-4342 ◽  
Author(s):  
S Camus ◽  
C Quevedo ◽  
S Menéndez ◽  
I Paramonov ◽  
P F W Stouten ◽  
...  
Keyword(s):  
High Throughput ◽  
Therapeutic Target ◽  
High Throughput Screening ◽  
Phosphorylase Kinase ◽  
Novel Therapeutic

scholarly journals High throughput screening of natural products for LDH inhibition as an anti‐mitotic therapeutic target (585.3)

2014 ◽  
Vol 28 (S1) ◽  
Author(s):  
Shihab Deiab ◽  
Elizabeth Mazzio ◽  
Najla Zarmouh ◽  
Nzinga Mack ◽  
Karam Soliman
Keyword(s):  
Natural Products ◽  
High Throughput ◽  
Therapeutic Target ◽  
High Throughput Screening

scholarly journals Multiplex Enzyme Assays and Inhibitor Screening by Mass Spectrometry

2010 ◽  
Vol 15 (8) ◽  
pp. 1001-1007 ◽  
Author(s):  
Rakesh Rathore ◽  
Patrick Pribil ◽  
Jay J. Corr ◽  
William L. Seibel ◽  
Artem Evdokimov ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Therapeutic Target ◽  
High Throughput Screening ◽  
Multiplex Assay ◽  
Enzyme Assays ◽  
Readout System ◽  
Inhibitor Screening ◽  
Serial Process ◽  
Pass Through

Current methods for high-throughput screening (HTS) use a serial process to evaluate compounds as inhibitors toward a single therapeutic target, but as the demand to reduce screening time and cost continues to grow, one solution is the development of multiplex technology. In this communication, the multiplex assay capability of a mass spectrometry (MS)–based readout system is verified using a kinase and esterase reaction simultaneously. Furthermore, the MS-based readout is shown to be compatible with a typical HTS workflow by identifying and validating several new inhibitors for each enzyme from a small library of compounds. These data confirm that it is possible to monitor inhibition of multiple therapeutic targets with one pass through the compound repository, thus demonstrating the potential for MS-based methods to become a method of choice for HTS of isolated enzymes.

scholarly journals Repurposing cytarabine for treating primary effusion lymphoma by targeting KSHV latent and lytic replications

2018 ◽  
Author(s):  
Marion Gruffaz ◽  
Shenghua Zhou ◽  
Karthik Vasan ◽  
Teresa Rushing ◽  
Qing Liu Michael ◽  
...  
Keyword(s):  
Mouse Model ◽  
High Throughput ◽  
High Throughput Screening ◽  
Therapeutic Agent ◽  
Potent Inhibitor ◽  
Primary Effusion Lymphoma ◽  
Viral Dna ◽  
Dna And Rna ◽  
Virion Production ◽  
Novel Therapeutic

AbstractOncogenic Kaposi’s sarcoma-associated herpesvirus (KSHV) is etiologically linked to primary effusion lymphoma (PEL), an aggressive and non-treatable malignancy commonly found in AIDS patients. In this study, we performed a high throughput screening of 3,731 characterized compounds, and identified cytarabine approved by FDA for treating numerous types of cancer as a potent inhibitor of KSHV-induced PEL. We showed the high efficacy of cytarabine in the growth inhibition of various PEL cells by inducing cell cycle arrest and apoptosis. Cytarabine inhibited host DNA and RNA syntheses and therefore induced cellular cytotoxicity. Furthermore, cytarabine inhibited viral DNA and RNA syntheses and induced the the rapid degradation of KSHV major latent protein LANA, leading to the suppression of KSHV latent replication. Importantly, cytarabine effectively inhibited active KSHV replication and virion production in PEL cells. Finally, cytarabine treatments not only effectively inhibited the initiation and progression of PEL tumors, but also induced regression of grown PEL tumors in a xenograft mouse model. Together, our study has identified cytarabine as novel therapeutic agent for treating PEL as well as eliminating KSHV persistent infection.ImportancePrimary effusion lymphoma is an aggressive malignancy caused by Kaposi’s sarcoma-associated herpesvirus. The outcome of primary effusion lymphoma is dismal without specific treatment. Through a high throughput screening of characterized compounds, we identified a FDA-approved compound cytarabine as a potent inhibitor of primary effusion lymphoma. We showed that cytarabine induced regression of PEL tumors in a xenograft mouse model. Cytarabine inhibited host and viral DNA and RNA syntheses, resulting in the induction of cytotoxicity. Of interest, cytarabine induced the degradation of KSHV major latent protein LANA, hence suppressing KSHV latent replication, which is required for PEL survival. Furthermore, cytarabine inhibited KSHV lytic replication program, preventing virion production. Our findings identified cytarabine as novel therapeutic agent for treating PEL as well as for eliminating KSHV persistent infection. Since cytarabine is already approved by the FDA, it might be an ideal candidate for repurposing for PEL therapy and for further evaluation in advanced clinical trials.

scholarly journals Assay Miniaturization for Ultra-High Throughput Screening of Combinatorial and Discrete Compound Libraries: A 9600-Well (0.2 Microliter) Assay System

1998 ◽  
Vol 3 (1) ◽  
pp. 55-62 ◽  
Author(s):  
Kevin R. Oldenburg ◽  
Ji-Hu Zhang ◽  
Tongming Chen ◽  
Anthony Maffia ◽  
Karl F. Blom ◽  
...  
Keyword(s):  
High Throughput ◽  
Combinatorial Chemistry ◽  
Therapeutic Target ◽  
High Throughput Screening ◽  
Detection System ◽  
Screening System ◽  
Handling System ◽  
Liquid Handling ◽  
Compound Libraries ◽  
Plate Design

Combinatorial chemistry has opened a new realm of chemical entities in the search for novel therapeutics. Combinatorial chemistry is currently adding hundreds of thousands of compounds to similar numbers available from years of synthesis by medicinal chemistry. It is not unreasonable to expect that over the next several years, nearly a million compounds will be available for screening against each therapeutic target. The number of potential targets will also be increasing with the advances in genomics. With the increasing number of compounds to be screened against an increasing number of targets, it is becoming increasingly difficult and costly to obtain the required amounts of key biological material needed to screen these compounds. One obvious solution is to miniaturize the assays so that the biological reagent supply doesn't need to increase. To this end, we have developed an ultra-high throughput screening system comprised of a new plate design (9600-well), detection system, and liquid handling system. This new format is capable of performing assays in as little as 0.2 Al. The results obtained from this system compare favorably to those obtained in the standard 96-well format.

scholarly journals High‐Throughput Screening Identifies Novel Therapeutic Targets for Merkel Cell Carcinoma

2019 ◽  
Vol 33 (S1) ◽  
Author(s):  
Khalid A. Garman ◽  
Tara B. Gelb ◽  
Daniel J. Urban ◽  
Matthew D. Hall ◽  
Isaac F. Brownell
Keyword(s):  
Cell Carcinoma ◽  
Merkel Cell Carcinoma ◽  
High Throughput ◽  
High Throughput Screening ◽  
Therapeutic Targets ◽  
Merkel Cell ◽  
Novel Therapeutic

Exploiting the Tr3 Interaction with Bcl-2 as a Novel Therapeutic Strategy of Cancer and Leukemia Therapy.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 256-256
Author(s):  
Dayong Zhai ◽  
Eduard Sergienko ◽  
Shinichi Kitada ◽  
Frederic Luciano ◽  
Arnold C. Satterswait ◽  
...  
Keyword(s):  
High Throughput ◽  
Conformational Changes ◽  
High Throughput Screening ◽  
Kinase Inhibitor ◽  
Leukemia Cell ◽  
Chemical Library ◽  
Over Expression ◽  
Amino Acid Peptide ◽  
Starting Point ◽  
Novel Therapeutic

Abstract TR3 (Nur77) is an orphan member of the retinoid/steroid family of nuclear receptors, which translocates from nucleus to mitochondria where it binds Bcl-2 and converts Bcl-2 from an anti-apoptotic to a pro-apoptotic protein (Li, et al. Science289: 1159, 2000; Lin, et al CELL116: 527, 2004). Because Bcl-2 is pathologically over-expressed in many ca cancers and leukemias, contributing to chemoresistance, we sought to exploit the TR3 interaction with Bcl-2 as a novel therapeutic approach. Mutagenesis studies identified a 9 amino-acid peptide from TR3 that mimics the effects of the full-length protein in terms of binding Bcl-2 and inducing conformational changes that covert Bcl-2 into a killer. A cell-permeable version of this TR3 peptide induced apoptosis of cultured leukemia cell lines (CEM; Jurkat), such that gene transfer-mediated over-expression of Bcl-2 enhanced sensitivity to the TR3 peptide. In contrast, Bcl-2 over-expression rendered these leukemia cells resistant to other inducers of apoptosis, such as the broad-spectrum kinase inhibitor Staurosporine. Mutant TR3 peptides that fail to bind Bcl-2 did not induce apoptosis. To identify chemical compounds that mimic the pro-apoptotic TR3 peptide, a Fluorescence Polarization Assay (FPA) was developed in which FITC-conjugated TR3 peptide binding to Bcl-2 or other appropriate anti-apoptotic Bcl-2-family proteins was measured in high-throughput mode. This high-throughput screening (HTS) assay performs well in 384 well format, with Z′ > 0.75. A chemical library of ~70,000 compounds was screened to identify compounds that displace the TR3 peptide, thus yielding hits that may serve as a potential starting point for drug discovery. Characterization of the properties of these compounds and their analogs will be described. (Supported by NIH grants R01-GM60554 and U54-HG003916).

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