1989 Fiscal Year Final Research Report Summary
Studies on the Structure and Synthesis of the Anesthetic Constituents of the Venoms of Solitary Wasps focused on Cyphononyx dorsalis L.
| Project/Area Number |
62470028
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
天然物有機化学
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| Research Institution | Shizuoka University |
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Principal Investigator |
OKUMURA Yasuaki SHIZUOKA UNIVERSITY, FACULTY OF SCIENCE, PROFESSOR, 理学部, 教授 (20021933)
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| Co-Investigator(Kenkyū-buntansha) |
IRIKAWA Hajime SHIZUOKA UNIVERSITY, FACULTY OF SCIENCE, ASSISTANT PROFESSOR, 理学部, 助教授 (40021943)
SAKURAI Atsushi SHIZUOKA UNIVERSITY, FACULTY OF SCIENCE, ASSISTANT PROFESSOR, 理学部, 助教授 (70021938)
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| Project Period (FY) |
1987 – 1989
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| Keywords | Cyphononyx dorsalis / Bioassay by Spinal Cord of Bullfrog / Bioassay by Hippocampal Neurons of Rat / Glutamate Antagonist / Neuroactive Peptides / Three-Dimensional Chromatogram |
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| Research Abstract |
Female Cyphononyx dorsalis L. were collected, and their venom sacs and glands were removed and extracted in two manners. The venom extracts were fractionated in two manners. The anesthetic constituents in the venom were searched with the aid of bioassay on neuroactivities examined by two methods: A) the effect of each fraction was studied on the isolated, intra-arterially perfused spinal cord of the bullfog; B) the effect of each fraction was studied by determination of cytosolic Ca^<2+> in hippocampal neurons of rats. The methods of extraction and fractionation were examined by the comparison of three-dimensional chromatograms obtained with a high-per-performance liquid chromatograph equipped with a multi-channel detector. Three-dimensional chromatograms indicated that the main components of the venom of Cyphononyx dorsalis are nucleosides and nucleotides. The venom extract prepared by irradiation of high-frequency followed by extraction with 0.02M acetic acid showed suppression of evoked potential in bioassay by the method A, and excitatory effects followed by weak action as glutamate antagonist in bioassay by the method B. This extract was fractionated by GPC into 12 fractions and gave four neuroactive peptides one of which was estimated to be Phe-Arg-Arg-Arg. Extraction with 50% acetonitrile-water and fracrionation of active fractions by reversed-phase partition chromatography gave adenosine, inosine, tryptophan, methyl 5-amino-2-picolinylaminopentanoate and a peptide regarded as cyclo-Lys-Ala-Ser-Thr. Synthetic studies on neuroactive peptides are under way.
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