1998 Fiscal Year Final Research Report Summary
Design of Gene Therapeutic Agent Possessing Cooperative DNA Alkylating Ability
| Project/Area Number |
09555283
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Synthetic chemistry
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
SUGIYAMA Hiroshi Tokyo Medical and Dental University, Institute for Medical and Dental Engineering, Professor, 医用器材研究所, 教授 (50183843)
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| Co-Investigator(Kenkyū-buntansha) |
SAITO Hiromitu Kyowa Hakko Kogyo Co., Ltd., Pharnacetical Research Laboratories, Chief Investig, 富士工場, 主査
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| Project Period (FY) |
1997 – 1998
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| Keywords | Antitumor agent / Heterodimer / Duocarmycin / Distamycin / DNA sequence / Molecular recognition |
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| Research Abstract |
Sequence-specific DNA alkylations have been a topic of much current interest due to their significant potential in molecular biology and human medicine. We developed general method to synthesize novel conjugates between segment A of Duocarmycin A (Du) and N-methylimidazole (lm)-N-methylpyrrole (Py) hairpin polyamides and their DNA alkylation. The conjugates PyPyPygammalmPyPyDu (8a) and JmPyPygammalmPyPyDu (8b) were designed to alkylate the target sequences (A/T)G(A/T)_2N(A/G) and (A/T)G(A/T)CN(A/G), respectively, according to Dervan's ring-pairing rule. High resolution denaturing gel electrophoresis indicated that 8a exclusively alkylated the A of the 5'-TGTAAAA-3' within a -400 bp DNA fragment Similarly, alkylation by 8b occurred exclusively at the G of the 5'-AGTCAGA-3' sequence with efficiency at nanomolar concentration. In order to better understand the structure of the alkylated DNA by these conjugates, the alkylation of non-self complementary duplex decanucleotides, ODNl and ODN2, were investigated. HPLC and ESMS analyses of the reaction of these ODNs with 8a and 8b demonstrated that both conjugates efficiently and selectively alkylate N3 of the purine bases of their target sequence.
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