Artemisia annua L. is a medicinal herb used in traditional Chinese medicine and the source of artemisinin, a component of a current leading antimalarial treatment. Traditionally, Artemisia annua tea has been used as a treatment for a variety of illnesses including malaria. However, artemisinin has been shown to inhibit P450 CYP3A4, which metabolizes approximately half of clinically used drugs today. Among those drugs are caffeine and acetaminophen, household staples that also inhibit CYP3A4 in great enough quantities. A previous MQP used Promega's P450-glo assays to analyze the interactions of those drugs with each other to determine, among other things, the safety of taking A. annua tea alongside either of the other two drugs. However, due to time and cost restraints, they were unable to analyze the full range of data necessary to make the proper analysis. In this MQP, we looked at using a 7-Benzyloxy-4-(trifluoromethyl) coumarin (BFC) based fluorescence assay in order to achieve similar results with a significantly reduced cost and equal convenience to the P45-glo assay. The first step was using the BFC assay to analyze the metabolism of BFC by CYP3A4 when in the presence of ketoconazole, a known CYP3A4 inhibitor, so as to independently confirm the assay’s consistency and suitability for use in the lab. Following that, the team used the assay to analyze A. annua tea, with the goal of performing assays on mixtures of A. annua tea with acetaminophen or caffeine, as the prior MQP did, but on a larger scale so as to confirm or deny their results using a wider range of concentrations. However, as we progressed through our investigation, we determined that the A. annua tea had an inherent fluorescence that was challenging the assay. By measuring the background fluorescence of the extract and subtracting it from the final fluorescence the assay was useful in analysis of CYP 3A4 activity in the BFC assay and may be useful in further studies subsequent to this project in measuring drug interactions with A. annua tea.

• This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

Creator

• Polansky, Ryan

Publisher

• Worcester Polytechnic Institute

Identifier

• E-project-042624-171400
• 121910

Advisor

• Scarlata, Suzanne Frances
• Weathers, Pamela

Year

• 2024

Date created

• 2024-04-26

Resource type

• Major Qualifying Project

Major

• Biology & Biotechnology

Source

• E-project-042624-171400

Rights statement

• In Copyright