Date of Graduation

5-2015

Document Type

Thesis

Degree Name

Bachelor of Science

Degree Level

Undergraduate

Department

Biological Sciences

Advisor

Shi, Wei Q

Reader

Lee, Sun-Ok

Second Reader

Du, Yuchun

Third Reader

Funkhouser, Eric M

Abstract

Ipomoeassin F is one member of a family of macromolecules isolated from the leaves of the morning glory flower, ipomoea squamosa, which has been used as a homeopathic cancer remedy for centuries (Zhu, Huang, Zheng, Zhu, & Yang, 2013). It has been proven to be highly cytotoxic to many cancer cell lines, but nothing is yet known of its mode of action in the cell (Cao, et al., 2007). To begin progress towards understanding the mode of action of ipomeoassin F, this study aimed to perform fluorescence imaging studies for the ultimate purpose of determining its cellular localization in MDA-MB-231 breast cancer cells. Attempts were made at both direct and indirect fluorescence imaging. To perform direct fluorescence imaging studies, two fluorescent dye probes known for their stability and ease of modification were synthesized: BODIPY and coumarin. The BODIPY synthesis was very challenging, and yield was very poor, averaging 0.46% yield. Both were subjected to SAR studies to evaluate their suitability for use in fluorescence imaging. Cytotoxicity assays revealed that neither dye probe can be used to study ipomoeassin F via direct fluorescence, because their addition to the ipomoeassin F molecule too severely inhibits its normal cytotoxic activity. To perform indirect fluorescence imaging studies, click chemistry was performed between a triazole coumarin fluorophore and a biorthogonal alkyne analog of ipomoeassin F to yield a fluorescent product. The reactions performed as part of this project were preliminary, and included a calibration titration, two model system reactions, and a study of the analog. These reactions elucidated conditions and suggestions that will be useful in future studies of ipomoeassin F.

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