Dissecting cellular processes using small molecules: identification of colchicine-like, taxol-like and other small molecules that perturb mitosis

Haggarty, Stephen J.; Mayer, Thomas; Miyamoto, David T.; Fathi, Reza; King, Randall W.; Mitchison, Timothy J.; Schreiber, Stuart L.

doi:10.1016/s1074-5521(00)00101-0

Cited by 238 publications

(117 citation statements)

References 37 publications

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“…Using this technology, we screened C. elegans for regenerative effects upon exposure to a chemical library enriched for compounds that may affect neurite outgrowth in mammalian cell cultures in vitro (21,22). The potential targets of the small-molecule library that we screened included various kinases, cytoskeletal proteins, endocytic vesicle trafficking components, and nuclear Red, control (valve) layer; yellow, flow layer; blue, immobilization layer.…”

Section: Resultsmentioning

confidence: 99%

Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration

Samara

Rohde

Gilleland

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

110

114

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Discovery of molecular mechanisms and chemical compounds that enhance neuronal regeneration can lead to development of therapeutics to combat nervous system injuries and neurodegenerative diseases. By combining high-throughput microfluidics and femtosecond laser microsurgery, we demonstrate for the first time largescale in vivo screens for identification of compounds that affect neurite regeneration. We performed thousands of microsurgeries at single-axon precision in the nematode Caenorhabditis elegans at a rate of 20 seconds per animal. Following surgeries, we exposed the animals to a hand-curated library of approximately one hundred small molecules and identified chemicals that significantly alter neurite regeneration. In particular, we found that the PKC kinase inhibitor staurosporine strongly modulates regeneration in a concentration-and neuronal type-specific manner. Two structurally unrelated PKC inhibitors produce similar effects. We further show that regeneration is significantly enhanced by the PKC activator prostratin. C. elegans | chemical screen | microfluidicsT he ability of neurons in the adult mammalian central nervous system to regenerate their axons after injury is extremely limited, which has been attributed to both extrinsic signals of the inhibitory glial environment (1) as well as intrinsic neuronal factors (2-4). The discovery of cell-permeable small molecules that modulate axon regrowth can potentiate the development of efficient therapeutic treatments for spinal cord injuries, brain trauma, stroke, and neurodegenerative diseases. Identification of such molecules can also provide valuable tools for fundamental investigations of the mechanisms involved in the regeneration process. Currently, small-molecule screens for neuronal regeneration are performed in simple in vitro cell culture systems. Such screens have already revealed large numbers of chemicals that enhance regeneration and/or affect cellular morphogenesis, yet many of these hits still remain untested in vivo. In addition, most in vitro studies do not translate to animal models and also fail to reveal off-target, toxic, or lethal effects. Thus, a thorough investigation of neuronal regeneration mechanisms requires in vivo neuronal injury models.In vivo investigation of neuronal regeneration has been performed mainly in mice and rats. However, their long developmental periods, complicated genetics and biology, and expensive maintenance prevent large-scale studies on these animals. The nematode Caenorhabditis elegans is a simple, well-studied, invertebrate model-organism with a fully mapped neuronal network comprising 302 neurons. Its short developmental cycle, simple and low-cost laboratory maintenance, and genetic amenability make it an ideal model for large-scale screens, rapid identification of the molecular targets of screened compounds, and discovery of novel signaling pathways implicated in regeneration.Until recently however, the small size of C. elegans (∼50 μm in diameter) prevented its use for investigation of neuronal re...

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Section: Resultsmentioning

confidence: 99%

Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration

Samara

Rohde

Gilleland

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

110

114

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“…The zebrafish-based screen was performed with a chemical library that was previously screened for mitotic inhibitors with cell lines (Haggarty et al, 2000). Of the 29 identified compounds affecting cell cycle activity in vivo, 14 compounds were novel.…”

Section: Exploring the Advantages Of In Vivo Screens Over Cellbased Amentioning

confidence: 99%

Simple vertebrate models for chemical genetics and drug discovery screens: Lessons from zebrafish and Xenopus

Wheeler

Brändli

2009

Developmental Dynamics

159

131

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Chemical genetics uses small molecules to modulate protein function and, in principle, has the potential to perturb any biochemical event in a complex cellular context. The application of chemical genetics to dissect biological processes has become an attractive alternative to mutagenesis screens due to its technical simplicity, inexpensive reagents, and low-startup costs. In vertebrates, only fish and amphibians are amenable to chemical genetic screens. Xenopus frogs share a long evolutionary history with mammals and so represent an excellent model to predict human biology. In this review, we discuss the lessons learned from chemical genetic studies carried out in zebrafish and Xenopus. We highlight how Xenopus can be employed as a convenient first-line animal model at various stages of the drug discovery and development process and comment on how they represent much-needed tools to bridge the gap between traditional in vitro and preclinical mammalian assays in biomedical research and drug development. Developmental

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“…Their structures have been confirmed by IR, 1 H NMR and mass spectral data. The compounds were tested for their anti-inflammatory activity.…”

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confidence: 79%

Synthesis and biological evaluation of some 4-(1H-indol-3-yl)-6-phenyl-1,2,3,4-tetrahydropyrimidin-2-ones/thiones as potent anti-inflammatory agents

Amir¹,

Javed²,

Kumar³

2008

Acta Pharmaceutica

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Among the wide range of heterocycles explored to develop pharmaceutically important molecules, pyrimidine has played an important role in medicinal chemistry. A survey of literature has shown that compounds having a pyrimidine nucleus possess a broad range of biological activities such as anticancer (1), antiviral (2), antibacterial (3), antimalarial (4), antihypertensive (5) and anti-inflammmatory activities (6, 7). Moreover, chemistry and synthesis of 1,2,3,4-tetrahydropyrimidin-2-ones/thiones have been attracting extensive attention. The importance of these tetrahydropyrimidines is mainly due to their close structural relationship to the clinically important dihydropyrimidine Twelve new 4-(1H-indol-3-yl)-6-phenyl-1,2,3,4-tetrahydropyrimidin-2-ones/thiones (7-18) have been synthesized by reacting 1-aryl-3-(1H-indol-3-yl)-2-propen-1-one with urea and thiourea in ethanolic potassium hydroxide. Their structures have been confirmed by IR, 1 H NMR and mass spectral data. The compounds were tested for their anti-inflammatory activity. Test results revealed that compounds showed 49.5 to 70.7% anti-inflammatory activity whereas the standard drug ibuprofen showed 86.4% activity at the same oral dose. Four compounds, 4-(1H-indol-3-yl)-6-(4-chlorophenyl)-1,2,3,4-tetrahydropyrimidin-2-one (8), 4-(1H-indol-3-yl)-6-(4-methylphenyl)-1,2,3,4-tetrahydropyrimidin-2-one (10), 4-(1H-indol-3-yl)-6-(4-chlorophenyl)-1,2,3,4-tetrahydropyrimidin-2-thione (14), 4-(1H-indol--3-yl)-6-(4-methylphenyl)-1,2,3,4-tetrahydropyrimidin-2-thione (16), that showed significant anti-inflammatory activity were selected to study their ulcerogenic and lipid peroxidation activities. All tested compounds showed significant reduction in the ulcerogenic potential and lipid peroxidation compared to the standard drug ibuprofen.

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Dissecting cellular processes using small molecules: identification of colchicine-like, taxol-like and other small molecules that perturb mitosis

Cited by 238 publications

References 37 publications

Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration

Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration

Simple vertebrate models for chemical genetics and drug discovery screens: Lessons from zebrafish and Xenopus

Synthesis and biological evaluation of some 4-(1H-indol-3-yl)-6-phenyl-1,2,3,4-tetrahydropyrimidin-2-ones/thiones as potent anti-inflammatory agents

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