Xiaoou WangFollow

Date of Award

2019

Document Type

Honors Thesis (Open Access)

Department

Colby College. Chemistry Dept.

Advisor(s)

Kevin Rice

Second Advisor

Julie Millard

Abstract

Laromustine is a chemotherapeutic sulfonylhydrazine prodrug used in clinical trials against acute myeloid leukemia. Though laromustine showed some success in clinical trials, more experiments are needed to understand the hematological toxicity and the molecular mechanisms of patients’ resistance. This project aims to develop a strategy to identify compounds synergistic with laromustine in cultured leukemia cells from a library of 450 FDA-approved compounds through a forward chemical genetic screen. To optimize the screen, the cell seeding density, doubling time, and dose response curves were determined. The optimized concentration of HL60 cells in these experiments was determined to be between 25,000 and 40,000 cells/well in 384 well-plates for 12 or 24 hr before the measurement. The LD50 of laromustine was determined to be 159 µM. A concentration of 50 µM was identified as optimal for the chemical genetic screen because when tested with temozolomide, a compound with a similar mechanism of action to laromustine, 50 µM laromustine and 500 µM temozolomide showed synergistic effects. Following those optimized conditions, the chemical genetic screen should have the potential to find compounds that enhance laromustine’s cytotoxicity, which, in the future, would help identify new molecular targets for laromustine’s mechanism of action.

Keywords

laromustine, chemical genetic screen

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