Highthroughput screening for small molecules with biological effects'

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High-throughput screening for small molecules
with biological effects. High throughput means
tens of thousands, even gt 100,000 tests per day.
Cyclosporine
FK506
rapamycin
taxol
vinblastine
Big (MW gt500) (natural products)
stereocenters Big here ltgt large
(macromolecules) Stereocenters asymmetric
carbons (4 diff. groups)
Small (MWlt500)
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Split pool combinatorial chemistry
Each bead really has many growing
molecules. All the molecules grown on a
given bead are the same.
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Diversity of molecular descriptors for different
chemical libraries (2 descriptors, of 138)
ACL natural compounds with known biological
activity they are more diverse even though fewer
in number
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Cytoblot technique for high-throughput
screening (gt tens of 1000s)
Also can screen for suppressors of Known drugs.
E.g., nocadozole blocks in mitosis, ? Phospho
signalnew drug that blocks signal, stops cells
before mitosis
E.g., anti-phospho-nucleolin,accumulates in
mitotic cells, So screen is for mitotis blockers
ECL enhanced chemiluminescence Enzyme
catalyzed light-producing reaction Enzyme is a
conjugate with a generic anti-Ab(blue).
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Automated image analysis for high-throughput
screening of cell morphology effects
E.g., neurite outgrowth from cultured neurons or
neuronal cell lines.
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Combining high-throughput small molecule
screening with genetically engineered cells to
connect ligands to proteins.
Add combinations of 6 oncogenes to normal human
diploid fibroblast to transform them into tumor
cells hTERT for immortalization SV40 large T
cell cycle checkpoint interference SV40 small T
cell cycle checkpoint interference Ras mutant
signal transduction control failure Human
papillloma virus (HPV) E6 inactivates p53 HPV E7
inactivates RB (retinoblastoma protein)
Screen for compounds that selectively inhibit the
growth of the transformed cells as opposed to the
parental isogenic normal cells.
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Not specific to an introduced gene
Topoisomerase Inhibitors And topos are
induced In the transformed cells (topoI or topoII)
Novel compound Kills via a non-apoptotic route
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Bush, E. Jens Fielitz, Lawrence Melvin, Michael
Martinez-Arnold, Timothy A. McKinsey, Ryan
Plichta,and Eric N. Olson. . A small molecular
activator of cardiac hypertrophy uncovered in a
chemical screen for modifiers of the calcineurin
signaling pathway. Proc Natl Acad Sci U S A 101,
2870-5 (2004).