| 研究課題/領域番号 |
19F19339
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| 研究機関 | 東京大学 |
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研究代表者 |
浦野 泰照 東京大学, 大学院薬学系研究科(薬学部), 教授 (20292956)
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| 研究分担者 |
KELLER SASCHA 東京大学, 薬学研究科(研究院), 外国人特別研究員
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| 研究期間 (年度) |
2019-11-08 – 2022-03-31
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| キーワード | Fluorescence / Cancer / Tumor / Rhodol / Spiro-cyclic / Computer-based |
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| 研究実績の概要 |
his project aims at finding suitable probes to detect various cancers by rational design of fluorescence probes based on a quantum chemical prediction of their intramolecular spiro-cyclization. Goals of the project include: (i) development of a computer-based model to predict properties of novel fluorescent molecules, (ii) the synthesis of a library of amino-acid and sugar derivatives of novel Si- or C-Rhodols that have promising properties; (iii) using those probes to visualize colon and stomach cancer as well as (iv) understanding of their overexpression of aminopeptidases and glycosidases in clinical specimens of cancerous colon and stomach tissue. Given the novel developed synthetic route which was even further enhanced fast and efficient synthesis of a huge variety of molecules is now possible. To develop a working model to predict the spiro-cyclization behavior of fluorescent probes, a library of several structurally different probes was synthesized, and their pKcycl-values were experimentally determined. After fine tuning of the in silica predicted values for said compounds the error margin was reduced to a satisfactory level. This should now allow us to predict the pKcycl-value for unknown molecules. Furthermore, the synthesis and the calculations could be expanded to C-Rhodols, which exhibit a different absorption and emission wavelength compared to the Si-Rhodols. This is important, since it would allow multi-color imaging of specimens.
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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
To calculate the important pKcycl-value of potential probes the free energy gap between the closed and the open form is estimated by a computational chemistry approach. After synthesizing a large library of compounds and determining their chemical properties, the calculations fit well with the experimental values. This allows us now to calculate different unknown compounds and select a suitable structure to be used as the core of our probes. Also, the mentioned pathway allows easy coupling to amino acids or sugars. Furthermore, C-Rhodols started to be of interest, too, hence a small library was already synthesized. Using C-Rhodols and Si-Rhodols in tandem would allow multi-color imaging. One core structure bound to a sugar-moiety showed a strong increase in fluorescence when tested in breast cancer specimen. Said probe will be tested in colon and stomach cancers. Furthermore, the developed synthesis allows easy coupling of an amino acid and a sugar to the same molecule, rendering it an AND-gate probe. Only when both groups have been cleaved a strong fluorescence is detectable. These probes have not been extensively studied yet and might be of great interest.
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| 今後の研究の推進方策 |
Since the calculations show satisfactory results, several possible structures are being calculated and the most suitable ones will be synthesized. In a straightforward manner, these probes will be bound to sugars or amino acids. The sugar or amino acid bound probes will then be tested on tumor specimen to find a suitable probe. Then the probes will be tested with purified enzymes but also with cancerous cell lines to verify their behavior in vitro. Since a small library of C-Rhodols has already been synthesized, we will also try to expand the strategy to these probes which show a different spectroscopic behavior. Necessary control experiments must be conducted to exclude false conclusions. Also, the AND-gate probes will be subject to explore their potential in cancer detection. The results of these experiments as well as the before mentioned synthetical pathway are subject to be published within next fiscal year.
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