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LC-MS-Based Screening of East Indian Sandalwood Oil (EISO) for Antitubercular and Antiplasmodial Mechanisms of Action
Poster Title: LC-MS-Based Screening of East Indian Sandalwood Oil (EISO) for Antitubercular and Antiplasmodial Mechanisms of Action
Submitted on 03 Apr 2017
Author(s): Thankhoe A. Rants’oa, Mansour Alturkia, Corey Levensonb, Angela I. Calderóna.
Affiliations: aDepartment of Drug Discovery and Development, Harrison School of Pharmacy, 4306B  Walker Building, Auburn University, Auburn, AL 36849. bSantalis Pharmaceuticals, 18618 Tuscany Stone, Suite 100, San Antonio, TX 78258
This poster was presented at PITTCON 2017
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Poster Information
Abstract: East Indian sandalwood oil (EISO) displayed antimycobacterial and antiplasmodial activities through unknown mechanisms. The goal of this study is to determine whether EISO exerts these by inhibiting enzymatic targets Mycobacterium tuberculosis shikimate kinase (MtSK) and Plasmodium falciparum thioredoxin reductase (PfTrxR) utilizing ESI-QTOF LC-MS-based functional assay approach. MtSK catalyzes the ATP-dependent phosphorylation of shikimate to shikimate-3-phosphate (S3P) in the shikimate pathway responsible for aromatic amino acids biosynthesis. PfTrxR catalyzes reduction of thioredoxin disulfide (Trx-S2) to thioredoxin dithiol (Trx-(SH)2) essential for antioxidant defense of the parasite. MtSK-inhibitory assay was carried out using a phenyl-hexyl 4.6x100mm,3.5 µm column with a gradient mobile phase of 0.1% formic acid (FA) in water and acetonitrile. Direct inhibition of S3P production was monitored through relative peak areas by LC-MS. EISO at 0.01% v/v in Tween-20 displayed 60% MtSK inhibition with IC50 of 0.015% v/v. EISO PfTrxR-inhibitory activity was evaluated using a Zorbax 300SB-C8 column, 2.1x100 mm,3.5 µm and a gradient mobile phase of 0.1% FA in water and 0.1% FA in acetonitrile. Inhibition of Trx-(SH)2 formation was monitored by LC-MS. To distinguish the two forms of thioredoxin, known major m/z values of multiply-charged protein envelope in the Trx-(SH)2 spectrum were deconvoluted to a product protein at a mass 11675.8711 Da and used for relative quantitation by displaying its extracted ion chromatogram. Major m/z values for the substrate Trx-S2 were deconvoluted at the mass 11673.9204 Da. The 0.01% EISO in Tween-20 showed 90% PfTrxR inhibition. This study reports LC-MS-based screening of EISO and its inhibitory activity on MtSK and PfTrxR.
Summary: This research displays LC-MS-based screening of East Indian Sandalwood oil for inhibitory activities on Mycobacterium tuberculosis shikimate kinase and Plasmodium falciparum thioredoxin reductase. It also shows the optimized sample preparation for stability in bioassay buffers and LC-MS analysis. References: 1. Diaz-Chavez ML et al. (2013). PLoS One, 8, e75053.
2. Fujisaki R et al. (2012). Southeast Asian Journal of Tropical Medicine and Public Health, 43, 270-279.
3. Misra BB and Dey S. (2012). Letters in Applied Microbiology, 55, 476-486.
4. Hammer KA et al. (1999). Journal of Applied Microbiology, 86, 446-452
5. Simithy J et al. (2014). Tuberculosis, 94, 152-158.
6. Munigunti R and Calderon AI. (2012). Rapid Communications in Mass Spectrometry, 26, 2051-2056.
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