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Journal of Glycobiology | ISSN: 2168-958X | Volume 7
Glycobiology & Glycoproteomics
5
th
International Conference on
&
August 27-28, 2018 | Toronto, Canada
Molecular Biology & Nucleic Acids
3
rd
International Conference on
Mechanistic investigations of α-glucosidase activity
Daniel C Hill
Emergent BioSolutions, USA
E
mergent BioSolutions is currently developing antivirals with an iminosugar structural motif that targets the endoplasmic
reticulum (ER) alpha-glucosidase enzymes. The mechanism of action is based on the inhibition of host glycosylation
pathway that leads to misfolded viral glycoproteins, resulting in reduced viral infectivity. Data generated by Emergent and
others indicate that iminosugars have the potential to be developed as a treatment for diseases such as Dengue Fever, influenza,
Ebola, and Zika. To support our drug discovery efforts, preliminary mechanistic investigations into the protein synthesis
and folding pathway have been initiated. We have verified the impact of inhibition on α-glucosidase activity by showing
inhibition of viral proliferation in human cell lines knocked out for each of the ER glucosidases using CRISPR-Cas9. With
collaborators at the Oxford Glycobiology Institute and a commercial MALDI-MS vendor, we evaluated in vitro glycan profiling
in the CRISPR cells by both a chemical derivatization/HPLC method and by MALDI-MS of permethylated glycans. These
orthogonal techniques confirm that knock-out of alpha-glucosidase 1 and alpha-glucosidase 2 enzymes prevents the cleavage
of the terminal glucose units involved in glycoprotein maturation. Examining the changes in glycan product distribution after
treatment of cell extracts with alpha-glucosidase 1 reveals the loss of a single glucose, while treatment with alpha-glucosidase
1 and alpha-glucosidase 2 results in the loss of two and three glucose units. These techniques can be used to confirm the
mechanism of action of new chemical entities from our iminosugar med-chem program. Work described here was performed
in collaboration with Anthony Treston and KellyWarfield of Emergent, DomAlonzi and Nicole Zitzmann of Oxford University
and Craig Day of Utah State University.
Biography
Daniel C Hill is a PhD chemist with 20 years’ experience in the pharmaceutical industry solving complex problems in results-driven departments working on pro-
grams in early discovery supporting candidate selection, drug development and on through to marketed commercial products. Dan has diverse experience in a
variety of therapy areas along the critical path of drug discovery and development including Neurokinin Receptor Antagonists, gamma-Secretase, 5HT-1b Receptor
Antagonists, Gap Junction Modulators, Topoisomerase Inhibitors, and Iminosugar alpha-Glucosidase Inhibitors.
hilld@ebsi.comDaniel C Hill, J Glycobiol 2018, Volume 7
DOI: 10.4172/2168-958X-C1-012