| Project/Area Number |
10672021
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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 |
Physical pharmacy
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| Research Institution | Nagasaki University |
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Principal Investigator |
KURODA Naotaka Nagasaki University, Professor, 薬学部, 教授 (50234612)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Mitsuhiro Nagasaki University, Assistant Professor, 薬学部, 助手 (40295093)
NAKASHIMA Kenichiro Nagasaki University, Professor, 大学院・薬学研究科, 教授 (30039656)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | arylboronic acid derivatives / lophine / chemiluminescence / horseradish peroxidase / photographic detection / enhancer / fluorescence / アリールボロン酸 / ペルオキシダーゼ / 糖 / ジオール |
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| Research Abstract |
Four kinds of novel arylboronic acids, which are the derivatives of lophine and its analogues, were synthesized based on the fact that lophine shows strong fluorescence (FL) and chemiluminescence (CL).
Since some arylboronic acids are known to enhance the luminol-hydrogen peroxide-horseradish peroxidase (HRP) CL reaction, we evaluated those as enhancers at first. Among the compounds examined, 4-[4,5-di(2-pyridyl)-1H-imidazol-2-yl]phenyl boronic acid (DPPA) was found to be the most potent enhancer. The detection limit of HRP with DPPA was 0.15 ng (ca. 3.5 fmol) which was ca. 180 times better than that in the absence of the enhancer. By using DPPA with the effect of prolonging the light emission, the CL system was applied to the photographic detection of HRP on a nitrocellulose membrane. After optimizing the CL reaction conditions, as little as 0.1 ng of HRP was photographically detectable. This method was successfully applied to the western blotting assay of anti-human T-cell leukemia virus type-I.
Next, FL and CL characteristics of the arylboronic acids were examined as fundamental studies on developing FL probes or FL/CL labeling reagents for diol compounds. 4-(4,5-Diphenyl-1H-imidazol-2-yl )phenylboronic acid (DPA) showed a stronger FL than lophine whereas the other derivatives had weaker FL. Stronger CL was not observed for all derivatives, compared to that of lophine. Although the reaction of DPA with some diols was estimated by TLC so far, further investigation on the reactivity and spectral studies will be required.
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