| 研究実績の概要 |
The main goal of the project is to find the active constituent of hijiki responsible for sphingomyelin synthase inhibition. So far, we have completed the methanol extraction of hijiki plant material in bulk and separated the active constituents based on solvent partition. All the solvent partitions are subjected to SMS inhibition screening assay using high-performance liquid chromatography with fluorescence detection. As consistent with our preliminary results, we found the strong SMS inhibition in hexane fraction, and it's further fractionated into several fractions. The SMS screening assay results further confirmed the active compound in fraction 3. The high-resolution mass spectrometry and NMR analysis was performed to reveal the structural information of the active compound.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Based on the analysis results, we judged that molecular weight of the compound is between 300-330,suggesting it as a small molecule inhibitor. Further, the active compound is a mixture of lipids belong to fatty acid family. As we found that the active constituent of hijiki is an inseparable isomeric mixture, there is delay in invitro experiments. We need to confirm the structure and purify the active constituent in order to reveal the in vitro actions of the hijiki derived SMS inhibitor. As a first effort, we made derivatization with a UV chromophore to separate the isomers using HPLC system. We are making all our efforts on the purification process. As soon as we accomplished it, the rest of the aims will be focused.
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| 今後の研究の推進方策 |
In the future, we will focus on the purification of active constituent using HPLC in order to confirm the structure of the major isomers. HRMS and NMR analysis will be formed. Upon obtaining the active constituent of hijiki in mg scale by HPLC purification, the SMS screening assay will be performed to know the inhibitory concentrations. Then the action of active constituent on lipid droplet accumulation in fatty liver cell model will be studied as described in the proposal aim 2.
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