高通量スクリーニングのためのフッ素-NMR実験:理論的側面,実践的検討,および適用範囲
Claudio Dalvit1, Paul E Fagerness, Daneen T A Hadden
1Chemistry Department, Pharmacia, Viale Pasteur 10, 20014 Nerviano (MI), Italy. claudio.dalvit@pharmacia.com
Journal of the American Chemical Society
|June 19, 2003
まとめ
フッ素-19検出を用いた競合のリガンドベースのNMRスクリーニングは,薬剤候補者を特定するための高通量方法を提供します. このFAXS技術は,混合物を効率的にスクリーニングし,薬物発見のための伝統的なNMRスクリーニングの限界を克服します.
科学分野:
- バイオケミストリー バイオケミストリー
- 化学生物学 化学生物学とは
- 構造生物学 構造生物学とは
背景:
- 従来のリガンドベースのNMRスクリーニングは,溶解性と親和性に関する課題に直面しています.
- 競争に基づくNMRスクリーニングは,標的結合分子を特定するための改善されたアプローチを提供します.
- フッ化物-19 (19F) の検出は,NMRスクリーニングにおける陽子 (1H) の検出よりも利点があります.
研究 の 目的:
- 新しい19FベースのコンペティションリガンドベースのNMRスクリーニング方法の理論的側面を提示し,議論する.
- FAXS (フッ素化学シフトアニソトロピーとスクリーニングのための交換) テクニックを詳細に説明します.
- 実験条件,検出限界,および結合定数測定を調査する.
主な方法:
- 19F検出によるコンペティションリガンドベースのNMRスクリーニングを使用しました.
- FAXS法で,フッ素の化学的シフトアニソトロピーと交換を活用した.
- 対象タンパク質 (ヒト血清アルブミン) に対して,基準リガンドの存在で,化学混合物をスクリーニングした.
主要な成果:
- 19F検出は,プロトンバッファを使用しても,大規模な化学混合物のスクリーニングを自動分析で可能にします.
- FAXS法では,限界溶解性と高い親和性を有する分子を特定することができます.
- スパイ分子選択によるNMRヒットに対するより低い結合親和度値を示した.
結論:
- FAXS方法は,限られた量のタンパク質を必要とする高速で高通量スクリーニング技術です.
- 19Fベースの競争性NMRスクリーニングは,従来の方法の限界を克服しています.
- FAXSは,確立された非NMRの高通量スクリーニング技術に対する効果的な代替手段として有望であることを示しています.
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01:07Nuclear Magnetic Resonance (NMR): Overview
Nuclear magnetic resonance (NMR) is a phenomenon exhibited by certain nuclei that can absorb characteristic radio frequency radiation under certain conditions. NMR has been extensively applied in molecular spectroscopy and medical diagnostic imaging. In both these applications, the molecule or subject under study is placed in a magnetic field and irradiated with radio frequency energy.
NMR spectroscopy generates a spectrum where the characteristic absorption frequencies of the sample are...
NMR spectroscopy generates a spectrum where the characteristic absorption frequencies of the sample are...
01:20NMR Spectrometers: Overview
NMR spectrometers consist of a strong magnet, a radiofrequency transmitter, and a detector attached to a computer console for recording spectra of samples containing NMR-active nuclei. In first-generation NMR instruments called continuous-wave spectrometers, the resonance frequencies of the nuclei are determined by frequency-sweep or field-sweep methods. The magnetic field strength is fixed and the rf signal is swept in the former, while the radiofrequency signal is fixed and the magnetic field...
01:12Two-Dimensional (2D) NMR: Overview
The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily interpreted using 2-dimensional (2D) NMR. Unlike 1D NMR, 2D NMR has two frequency axes that provide the coupling information between the nucleus A and nucleus B in a molecule. The process from which 2D spectra are obtained has four steps.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse.
01:17NMR Spectrometers: Radiofrequency Pulses and Pulse Sequences
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01:25Applications Of NMR In Biology
Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
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01:182D NMR: Overview of Heteronuclear Correlation Techniques
Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds' structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other axis.