5 research outputs found
Assembly of Spirooxindole Derivatives via Organocatalytic Iminium-Enamine Cascade Reactions
The assembly of complex spirocyclopentaneoxindoles via a novel organocatalytic iminium-enamine cascade process is reported. Reactions between 3-substituted oxindoles and α,β-unsaturated aldehydes catalyzed by second generation prolinol ethers provided the desired products in high yield with excellent levels of enantioselectivity in a single step
Chemoproteomics-Enabled Covalent Ligand Screening Reveals ALDH3A1 as a Lung Cancer Therapy Target
Chemical
genetics is a powerful approach for identifying therapeutically
active small molecules, but identifying the mechanisms of action underlying
hit compounds remains challenging. Chemoproteomic platforms have arisen
to tackle this challenge and enable rapid mechanistic deconvolution
of small-molecule screening hits. Here, we have screened a cysteine-reactive
covalent ligand library to identify hit compounds that impair cell
survival and proliferation in nonsmall cell lung carcinoma cells,
but not in primary human bronchial epithelial cells. Through this
screen, we identified a covalent ligand hit, DKM 3-42, which impaired
both <i>in situ</i> and <i>in vivo</i> lung cancer
pathogenicity. We used activity-based protein profiling to discover
that the primary target of DKM 3-42 was the catalytic cysteine in
aldehyde dehydrogenase 3A1 (ALDH3A1). We performed further chemoproteomics-enabled
covalent ligand screening directly against ALDH3A1, and identified
a more potent and selective lead covalent ligand, EN40, which inhibits
ALDH3A1 activity and impairs lung cancer pathogenicity. We show here
that ALDH3A1 represents a potentially novel therapeutic target for
lung cancers that express ALDH3A1 and put forth two selective ALDH3A1
inhibitors. Overall, we show the utility of combining chemical genetics
screening of covalent ligand libraries with chemoproteomic approaches
to rapidly identify anticancer leads and targets
Chemoproteomics-Enabled Covalent Ligand Screening Reveals ALDH3A1 as a Lung Cancer Therapy Target
Chemical
genetics is a powerful approach for identifying therapeutically
active small molecules, but identifying the mechanisms of action underlying
hit compounds remains challenging. Chemoproteomic platforms have arisen
to tackle this challenge and enable rapid mechanistic deconvolution
of small-molecule screening hits. Here, we have screened a cysteine-reactive
covalent ligand library to identify hit compounds that impair cell
survival and proliferation in nonsmall cell lung carcinoma cells,
but not in primary human bronchial epithelial cells. Through this
screen, we identified a covalent ligand hit, DKM 3-42, which impaired
both <i>in situ</i> and <i>in vivo</i> lung cancer
pathogenicity. We used activity-based protein profiling to discover
that the primary target of DKM 3-42 was the catalytic cysteine in
aldehyde dehydrogenase 3A1 (ALDH3A1). We performed further chemoproteomics-enabled
covalent ligand screening directly against ALDH3A1, and identified
a more potent and selective lead covalent ligand, EN40, which inhibits
ALDH3A1 activity and impairs lung cancer pathogenicity. We show here
that ALDH3A1 represents a potentially novel therapeutic target for
lung cancers that express ALDH3A1 and put forth two selective ALDH3A1
inhibitors. Overall, we show the utility of combining chemical genetics
screening of covalent ligand libraries with chemoproteomic approaches
to rapidly identify anticancer leads and targets
Chemoproteomics-Enabled Covalent Ligand Screening Reveals ALDH3A1 as a Lung Cancer Therapy Target
Chemical
genetics is a powerful approach for identifying therapeutically
active small molecules, but identifying the mechanisms of action underlying
hit compounds remains challenging. Chemoproteomic platforms have arisen
to tackle this challenge and enable rapid mechanistic deconvolution
of small-molecule screening hits. Here, we have screened a cysteine-reactive
covalent ligand library to identify hit compounds that impair cell
survival and proliferation in nonsmall cell lung carcinoma cells,
but not in primary human bronchial epithelial cells. Through this
screen, we identified a covalent ligand hit, DKM 3-42, which impaired
both <i>in situ</i> and <i>in vivo</i> lung cancer
pathogenicity. We used activity-based protein profiling to discover
that the primary target of DKM 3-42 was the catalytic cysteine in
aldehyde dehydrogenase 3A1 (ALDH3A1). We performed further chemoproteomics-enabled
covalent ligand screening directly against ALDH3A1, and identified
a more potent and selective lead covalent ligand, EN40, which inhibits
ALDH3A1 activity and impairs lung cancer pathogenicity. We show here
that ALDH3A1 represents a potentially novel therapeutic target for
lung cancers that express ALDH3A1 and put forth two selective ALDH3A1
inhibitors. Overall, we show the utility of combining chemical genetics
screening of covalent ligand libraries with chemoproteomic approaches
to rapidly identify anticancer leads and targets
Protein Delivery Using Cys<sub>2</sub>–His<sub>2</sub> Zinc-Finger Domains
The
development of new methods for delivering proteins into cells
is a central challenge for advancing both basic research and therapeutic
applications. We previously reported that zinc-finger nuclease proteins
are intrinsically cell-permeable due to the cell-penetrating activity
of the Cys<sub>2</sub>–His<sub>2</sub> zinc-finger domain.
Here, we demonstrate that genetically fused zinc-finger motifs can
transport proteins and enzymes into a wide range of primary and transformed
mammalian cell types. We show that zinc-finger domains mediate protein
uptake at efficiencies that exceed conventional protein transduction
systems and do so without compromising enzyme activity. In addition,
we demonstrate that zinc-finger proteins enter cells primarily through
macropinocytosis and facilitate high levels of cytosolic delivery.
These findings establish zinc-finger proteins as not only useful tools
for targeted genome engineering but also effective reagents for protein
delivery