The study of oligonucleotide block synthesis on a support
| Project/Area Number |
59870069
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemical pharmacy
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| Research Institution | Osaka University |
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Principal Investigator |
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| Project Period (FY) |
1984 – 1985
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| Project Status |
Completed (Fiscal Year 1985)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1985: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1984: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | triester method / polymer support / anisidate linkage / capping reaction / 亜硝酸イソアミル |
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| Research Abstract |
1. The synthesis of nucleotide resin To synthesize the nucleotide resin with P-N linkage which can be removea by the treatment of isoamylnitrite in neutral condition, 5'-O-(DMTr)thymidine-3'-ochlorophenylphosphate was reacted with trichloroethyl ester of 4-aminophenyl acetate. The resultant derivative was joined to the polystyrene resin of 1% divinylbenzene. The loaded protected thymidine was 97% <micro> mol per 1 g of the resin. Such resin was treated with the excess of isoamylnitrite in pyridineacetic acid at r.t. to release the nucleotidic compound from the resin. It was found that the mononucleotide was released quantitatively after 3 hrs treatment with isoamylnitrite.
2. The condition of capping The tetramer (d-AAAT) was synthesized by the phosphotriester method on the above mentioned resin. However, the condensation yield was low between 16-70 % except the first condensation. The reason was found that acetic anhydride was reacted with the nitrogen on P-N linkage in the presence dimethylaminopyridine (DMAP) as catalyst, and the resultant unstable linkage was cleaved immediately. But it was also found that the acetylation was not occurred on the nitrogen if the DMAP was not used. In this condition, the P-N linkage was stable. Next, we studied the condition of acetylation with acetic anhydride in the absence of DMAP. Then it was found that the acetylation was completed at r.t. for 1 hr when 4 M of acetic anhydride in pyridine was used. 3. The synthesis of oligonucleotides
A hexanucleotide (d- <T_6> ) and dodecanucleotide (d-CTATGTATCCCT) were synthesized by the phosphotriester method on the above mentioned resin and the modified capping condition. After the syntheses were finished, the resins were treated with isomylnitrite, pyridine-aldoxmate and conc. ammonia. The unblocked hexamer and dodecamer which still have 3'-o-chlorophenylphosphate were isolated by the reversed phase silica gel high pressure liquid chromatography.
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Report
(1 results)
Research Products
(6 results)
