1997 Fiscal Year Final Research Report Summary
Confromational analysis of fluorovitamin D analogs interacting with vitamin D receptor using ^<19>F-NMR spectroscopy
| Project/Area Number |
08672415
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemical pharmacy
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
SHIMIZU Masato Tokyo Medical and Dental University, Institute for Medical and Dental Engineering, Associate Professor, 医用器材研究所, 助教授 (50126231)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Keiko Tokyo Medical and Dental University, Institute for Medical and Dental Engineerin, 医用器材研究所, 助手 (90147017)
YAMADA Sachiko Tokyo Medical and Dental University, Institute for Medical and Dental Engineerin, 医用器材研究所, 教授 (10014078)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Fluorovitamin D analog / Active vitamin D / Vitamin D receptor / Vitamin D-receptor complex / ^<19>F-NMR / Active conformation / Biological expression mechanism |
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| Research Abstract |
The active vitamin D induces biological responses via genomic michanism by binding to the vitamin receptor (VDR) which mediates gene expression. Formation of vitamin D-VDR complex is the first step for expressing of biological actions and the three-demensional structure of the active vitamin D at the time of interacting with VDR has not been determined up to now. To directly examine a strereostructure of vitamin D in the VDR-ligand complex by ^<19>F-NMR spectroscopy, we designed the fluorinated vitamin D analogs as probe compounds.
We successfully synthesized 4,4-difluorovitamin D and 19-fluorovitamin D analogs as probes to monitor the A-ring and the conjugated triene conformations of vitamin D,respectively. Binding affinity of 4,4-difluorovitamin D and 19-fluorivitamin D analog to bovine thymus VDR was about 1/100 and 1/10, respectively, as effective as the active vitamin D.To analyze the A-ring and the conjugated triene conformational equilibria, we investigated the ^<19>F-NMR spectra of the fluorovitamin D analogs at a variable temperature. Each of these compounds gave separate signals at below-60ﾟC,for the tworing A conformations. These analogs may serve as excellent probes for the A-ring conformational analysis.
The rat vitamin D receptor ligand binding domain (VDR-LBD) was expressed as fusion proteins in bacteria. The ^<19>F-NMR spectrum of VDR-LBD-fluorovitamin D complex represented additional signals from bound ligand. We have begun a systematic investigation of the assignment of the ligand conformation in the bound state.
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