Elucidation and Identification of Endogenous Benzodiazepine and Elucidation of Biological Properties of the Benzodiazepine
| Project/Area Number |
01571259
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
応用薬理学・医療系薬学
|
|---|
| Research Institution | Tokyo University of Science |
|---|
Principal Investigator |
KUBOTA Kazuhiko Science University of Tokyo, Department of Pharmaceutical Sciences, Professor, 薬学部, 教授 (30084411)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
HAZATO Tadahiko The Tokyo Metropolitan Institute of Medical Science, Department of Chemotherapy,, 化学療法研究部, 主任研究員 (60109949)
|
|---|
| Project Period (FY) |
1989 – 1990
|
| Project Status |
Completed (Fiscal Year 1991)
|
| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥1,200,000 (Direct Cost: ¥1,200,000)
|
|---|
| Keywords | Benzodiazepine / Receptor / Endogenous Ligand / Bovine Brain / Bovine Spinal Cord / Isolation / 牛脊随 / 精製 / 牛背髄 |
|---|
| Research Abstract |
In this study, we aimed to isolate and identify the substances that have been expected to exist in the central nervous system and bind selectively to the benzodiazepine receptor in the brain.
In order to conferm if endogenous benzodiazepine ligands isolated will bind to the benzodiazepine receptor, we obtained a fraction containing benzodiazepine receptor from the mice brain. Radioactive flunitrazepam labled with tritium was bound to the fraction, and replaced with fractions that were isolateed from the central nervous system and contained endogenous benzodiazepine ligands. Determination of endogenous benzodiazepine ligands was made through the degree of displacement of flunitrazepam by the endogenous ligands.
1)Search of endogenous ligands in the bovine brain
The bovine brain cortex was homogenized and extracted with acetone-methanol mixture. Detectable, but insufficient amount of endogenous ligands was obtained from this procedure.
2)Isolation of endogenous ligands from the bovine spinal cord.
The homogenate of the bovine spinal cord was subjected to the isolation of ligands through gel filtration and other procedures, resulting in the confirmation of endogenous benzodiazepine ligands having high affinity to benzodiazepine receptor preparation in the elutes of fraction number 47 to 49. The endogenous ligands showed their molecular weights to be less than 1, 000 and to be other substances than peptide and than diazepam, desmethyl diazepam and methy b-carboline ester. In addition, they were composed of more than 5 substances having different molecular sizes.
3)Isolation of ligands from the monkey brain and human serum
We tried to detect endogenous ligands from these sources by using similar methods as those for bovine spinal cord, but failed to isolate endogenous ligands.
|
Report
(3 results)
Research Products
(8 results)
