2023 Fiscal Year Research-status Report
Investigation of FFAR activity by polyunsaturated fatty acid library constructed by solid-phase synthesis
| Project/Area Number |
23KJ0751
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SHI YAOHONG 東京大学, 工学系研究科, 特別研究員(DC1)
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| Project Period (FY) |
2023-04-25 – 2026-03-31
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| Keywords | fatty acid / FFAR1 / FFAR4 / solid-phase synthesis / PUFA / GPCR |
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| Outline of Annual Research Achievements |
In the past year, I have synthesized several PUFA analogs. These efforts have led to the successful synthesis of a series of innovative PUFA analogs with unique characteristics. According to the normal PUFAs, the following compounds were synthesized, most have no commercial sources: PUFAs with different double bond numbers from natural PUFAs as well as PUFAs with an odd number of carbon chains.
Furthering the pursuit of innovation, the synthesis expanded beyond the conventional PUFA structures, that is those of omega-3 or omega-6 fatty acids. In addition to these PUFAs, I also finished the synthesis of omega-2 and omega-1 PUFAs, which are rarely obtained from commercial suppliers. In the future, the biochemistry functions will be under discussion.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
This project has been progressing smoothly as planned.
It took approximately three months to optimize the current synthesis method and go through the standard synthesis process for all the PUFA analogs. After initially synthesizing several PUFAs, I encountered challenges with compound purification and purity determination, which required more effort than anticipated. However, after testing various experimental conditions, suitable methods for the post-processing steps were identified, and the target compounds were successfully obtained. Although these steps required additional time, they did not significantly hinder the overall progress.
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| Strategy for Future Research Activity |
The small PUFA-derived library has now been established with about 20 new PUFAs, most of which have no commercial sources.
In the future, the biochemical functions of these PUFAs will be initially studied.
The first assay method under consideration is the TGF-α shedding assay. By using this method, the activity of FFAR1 and FFAR4 will be screened in the synthesized PUFA-derived library. Based on the data obtained, appropriate analytical methods will be applied to assess different factors. Moreover, biased agonism will be investigated using traditional second-messenger assays, such as the cAMP accumulation assay or the Ca2+ mobilization assay, both of which offer high sensitivity.
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| Causes of Carryover |
In the coming year, most of the funds will be allocated to purchasing reagents for biological experiments. With the majority of the synthesis now complete, I will shift my focus to studying the biochemical properties of the compounds. For this purpose, I will purchase various cell-related reagents and kits, which are more expensive than the synthesis reagents. Additionally, I will conduct the re-synthesis of PUFA analogs for which the current quantities are insufficient.
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