J. Ingram - 200743868 - Emerging Topics - Prof Nelson - Paper Presentation

Emerging Topics – Prof Nelson – J. Ingram

MJ Evans, A Saghatelian, EJ Sorensen, BF Cravatt

Introduction: Desired Phenotype

Evans et al wanted to find compounds that had an anti-proliferative effect through covalent bonding, on the human breast cancer line MDA-MB-231.

http://www.sciencephoto.com/media/252223/enlarge http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=HTB-26&Template=cellBiology

Introduction: Library Design

Covalent bonding ability.

Variable binding R group.

“Handle” for ease of elucidation.

Central core molecule used in this paper. Note spiro-epoxide in red, alkyne handle in

blue, and variable R group in purple.

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“Click” chemistry, used to connect the protein-drug complex to a reporter tag.

Various spiro-epoxide anticancer treatments: FR901464, Luminicin D and (±)-Fumagillin.

Introduction: Library Creation

MJE3

Testing: XTT Assay for Cell Proliferation

One compound found to slow the growth of MDA-MB-231 – MJE3.

Control = just solvent (DMSO)

Testing: MJE3 Target

MJE3 was found to uniquely label a protein, PGAM1

MJE3 Azide derived Rhodamine/Biotin tags

Testing: MJE3 Structure/Activity and Potential Pro-Drug

Tested spiro-epoxide activity via alkene derivative

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In situ vs. In vitro ability: MJE3 only works in living cells.

Methyl-ester variant, MJE52 shows activity in live cells too.

Flaws With the Paper

It would be very useful to determine the exact structure of PGAM1 complexed with MJE3 by X-ray crystallography.

This would give a better understanding of the molecule activation process, and allow the binding pocket to be explored, which could lead to an improved structure to be designed.

http://www.rcsb.org/pdb/results/results.do?outformat=&qrid=10B48E19&tabtoshow=Current

Future Work

Increased knowledge of activity based protein profiling.

Used in various cancer treatments

Finding known and novel targets for therapeutics

http://www.scripps.edu/cravatt/research.html

Conclusion

Created the first library where covalent binding was a privileged structure.

Found a structure which could significantly reduce the proliferation of aggressive breast cancer.