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Design and Synthesis of Firefly Luciferin Analogues to realize highly bright near infrared bioluminescence emission

萤火虫荧光素类似物的设计与合成实现高亮近红外生物发光

基本信息

批准号：
19J12615
负责人：
池田裕真
金额：
$ 1.09万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2019
资助国家：
日本
起止时间：
2019-04-25 至 2021-03-31
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-19J12615/
关键词：
生物発光イメージング近赤外ルシフェリンルシフェラーゼ

项目摘要

生体深部をイメージング可能なホタル生物発光系の構築を目的とし，発光反応中間体に注目した発光輝度の向上可能性を検証した．ATPと誘導体との縮合体は高輝度化に成功したものの，安定性や精製手法の確立，単離収率における課題が明らかとなった．そこで，基質化学構造の改変による長波長化および発光輝度双方の達成を試みることとした．開発指針としてホタル生物発光基質を有機蛍光色素と捉え，有機蛍光色素にみられる構造改変手法を模倣した．１.環融合戦略を基盤とした近赤外発光ホタルルシフェリン誘導体群の創製近赤外領域の光は，水などの生体分子による阻害が少ないことから，生体イメージングにおいて望ましいとされる．本研究では，既存の近赤外誘導体を基盤とし，環状アミン構造を組み込んだ計６種類の誘導体NIRLucsを合成した．いずれも天然のホタルルシフェラーゼFlucによる酵素認識を受け，近赤外発光を示した．さらに，NIRLucsは細胞膜透過性を持ち，生細胞系においても近赤外発光を示すことが確認された．良好な発光活性・血中滞留性を示したNIRLuc2を皮下腫瘍担癌マウスモデルへ応用したところ，天然の基質と比べて約７倍（近赤外では１６倍）の発光輝度を示した．２.環状アミノ基アゼチジン置換による発光輝度向上の可否検証ホタル発光基質をPush-Pull型の蛍光色素と見立て，Twisted Intramolecular Charge Transfer（TICT）の抑制による発光輝度向上の可否について検証した．本研究では，異なるルシフェリンコアに対して，アゼチジニル基を導入した誘導体を計５種類合成し，ルシフェリンにおけるアゼチジニル基置換効果を評価した．このうちアゼチジニル基を導入したA-1は，ジメチルアミノ基をもつ誘導体D-1と比較して，TICTの抑制により狙い通り蛍光量子収率・生物発光輝度共に向上することに成功した．

In the deep part of the body, it is possible to determine the biological light system, the light intensity, the upward possibility, the light intensity, the light intensity, the light Based on the chemical reaction, the photoluminescence of the photoluminescence system has been changed into a photoluminescence photoluminescence system on both sides of the biophoreum. it is indicated that the biological photoluminescence is used for the detection of light pigments and the modification of light pigments is simulated. 1. In the field of near-infrared light, water molecules are used to prevent the damage, and the body is not safe. In this study, the results show that there is a lot of light in the field of light. The production of environmental materials has been reported in recent years. The synthesis of NIRLucs has been carried out in a variety of organisms. The enzyme enzyme in Fluc has been detected, and it has been shown by near-infrared photoluminescence. The cell membrane permeability of NIRLucs has been detected. The cell line of the cell line was exposed to near-infrared light, and the light intensity was confirmed. Good photoactivity showed that the retention of NIRLuc2 in blood showed that cancer was burdened under the skin, and that the natural substrate was about 7 times higher than that of the whole body. 2. Is it possible to increase the luminosity of light-based Push- Pull phosphor pigments, Twisted Intramolecular Charge Transfer (TICT) suppresses the luminescence of photoluminescence, and increases the luminosity of photoluminescence? in this study, the system is used to determine whether it is possible to increase the light intensity of the photoluminescence. In the first place, we did not know what to do. In the first place, we did not know if there was a problem. In this way, we did not know what to do. In this way, we do not know what to do. In this way, we do not know what to do in the first place. In this way, we have a lot of information on the basis of this, and we have a lot of information about how to make a success in the first place. In this way, we have a lot of information about how to achieve success.