Allied Journal of Medical Research

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Allied Journal of Medical Research 44 7897 074717

Online Journals Of Drug Discovery

Drug discovery strategies are needed which will rapidly exploit multiple therapeutic targets related to the complex organic phenomenon changes that characterize a polygenic disorder like cancer. We report a replacement cell-based high-throughput technology for screening chemical libraries against several potential cancer target genes in parallel. Multiplex gene MGE) analysis provides direct and quantitative measurement of multiple endogenous mRNAs employing a multiplexed detection system coupled to reverse transcription-PCR. A multiplex assay for 6 genes overexpressed in cancer cells was wont to screen 9000 chemicals and known drugs within the human prostatic adenocarcinoma cell line PC-3. Active compounds that modulated organic phenomenon levels were identified, and IC50 values were determined for compounds that bind DNA, cell surface receptors, and components of intracellular signaling pathways. a category of steroids associated with the cardiac glycosides was identified that potently inhibited the cell wall Na+K+-ATPase leading to the inhibition of 4 of the prostate target genes including transcription factors Hoxb-13, hPSE/PDEF, hepatocyte nuclear factor-3α, and therefore the inhibitor of apoptosis, survivin. Representative compounds selectively induced apoptosis in PC-3 cells compared with the nonmetastatic cell line BPH-1. The multiplex assay distinguished potencies among structural variants, enabling structure-activity analysis suitable for chemical optimization studies. A second multiplex assay for five toxicological markers, Hsp70, Gadd153, Gadd45, O6-methylguanine-DNA methyltransferase, and cyclophilin, detected compounds that caused DNA damage and cellular stress and was a more sensitive and specific indicator of potential toxicity than measurement of cell viability. MGE analysis facilitates rapid drug screening and compound optimization, the simultaneous measurement of toxicological end points, and gene function analysis.

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