Abstract
A convergent synthetic approach was used to conjugate 2′,5′- oligoadenylate (2-5A, p5′A2′ [p5′A2′] np5′A) to phosphorodiamidate morpholino oligomers (morphants). To provide requisite quantities of 2-5A starting material, commercially and readily available synthons for solid-phase synthesis were adapted for larger scale solution synthesis. Thus, the tetranucleotide 5′- phosphoryladenylyl(2′→5′)adenylyl-(2′→5′) adenylyl(2′→5′)adenosine (p5′A2′p5′A2′] 2p5′A2′, tetramer 2-5A, 9) was synthesized starting with 2′,3′-O-dibenzoyl-N6,N6-dibenzoyl adenosine prepared from commercially available 5′-O-(4-monomethoxytrityl) adenosine. Coupling with N6-benzoyl-5′-O-(4,4′-dimethoxytrityl)- 3′-O-(tert-butyldimethylsilyl) adenosine-2′-(N,N-diisopropyl-2- cyanoethyl)phosphoramidite, followed by oxidization and deprotection, generated 5′-deprotected dimer 2-5A. Similar procedures lengthened the chain to form protected tetramer 2-5 A. The title product 9 p5′A(2′p5′A) 3 (tetramer 2-5A) was obtained through phosphorylation of the terminal 5′-hydroxy of the protected tetramer and removal of remaining protecting groups using concentrated ammonium hydroxide-ethanol (3:1, v/v) at 55°C and tetrabutylammonium fluoride (TBAF) in THF at room temperature, respectively. The 2-5A-phosphorodiamidate morpholino antisense chimera 11 (2-5A-morphant) was synthesized by covalently linking an aminolinker- functionalized phosphorodiamidate morpholino oligomer with periodate oxidized 2-5A tetramer (p5′A2′[p5′A2′]2p5′A). The resulting Schiff base was reduced with cyanoborohydride thereby transforming the ribose of the 2′-terminal nucleotide of 2-5A N-substituted morpholine. RNase L assays demonstrated that this novel 2-5A-antisense chimera had significant biological activity, thereby providing another potential tool for RNA ablation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 383-390 |
| Number of pages | 8 |
| Journal | Bioconjugate Chemistry |
| Volume | 16 |
| Issue number | 2 |
| DOIs | |
| State | Published - Mar 2005 |
Fingerprint
ASJC Scopus subject areas
- Chemistry(all)
- Organic Chemistry
- Clinical Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
Cite this
Endowing RNase H-inactive antisense with catalytic activity : 2-5A-morphants. / Zhou, Longhu; Civitello, Edgar R; Gupta, Nidhi; Silverman, Robert H.; Molinaro, Ross J.; Anderson, David E.; Torrence, Paul F.
In: Bioconjugate Chemistry, Vol. 16, No. 2, 03.2005, p. 383-390.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Endowing RNase H-inactive antisense with catalytic activity
T2 - 2-5A-morphants
AU - Zhou, Longhu
AU - Civitello, Edgar R
AU - Gupta, Nidhi
AU - Silverman, Robert H.
AU - Molinaro, Ross J.
AU - Anderson, David E.
AU - Torrence, Paul F.
PY - 2005/3
Y1 - 2005/3
N2 - A convergent synthetic approach was used to conjugate 2′,5′- oligoadenylate (2-5A, p5′A2′ [p5′A2′] np5′A) to phosphorodiamidate morpholino oligomers (morphants). To provide requisite quantities of 2-5A starting material, commercially and readily available synthons for solid-phase synthesis were adapted for larger scale solution synthesis. Thus, the tetranucleotide 5′- phosphoryladenylyl(2′→5′)adenylyl-(2′→5′) adenylyl(2′→5′)adenosine (p5′A2′p5′A2′] 2p5′A2′, tetramer 2-5A, 9) was synthesized starting with 2′,3′-O-dibenzoyl-N6,N6-dibenzoyl adenosine prepared from commercially available 5′-O-(4-monomethoxytrityl) adenosine. Coupling with N6-benzoyl-5′-O-(4,4′-dimethoxytrityl)- 3′-O-(tert-butyldimethylsilyl) adenosine-2′-(N,N-diisopropyl-2- cyanoethyl)phosphoramidite, followed by oxidization and deprotection, generated 5′-deprotected dimer 2-5A. Similar procedures lengthened the chain to form protected tetramer 2-5 A. The title product 9 p5′A(2′p5′A) 3 (tetramer 2-5A) was obtained through phosphorylation of the terminal 5′-hydroxy of the protected tetramer and removal of remaining protecting groups using concentrated ammonium hydroxide-ethanol (3:1, v/v) at 55°C and tetrabutylammonium fluoride (TBAF) in THF at room temperature, respectively. The 2-5A-phosphorodiamidate morpholino antisense chimera 11 (2-5A-morphant) was synthesized by covalently linking an aminolinker- functionalized phosphorodiamidate morpholino oligomer with periodate oxidized 2-5A tetramer (p5′A2′[p5′A2′]2p5′A). The resulting Schiff base was reduced with cyanoborohydride thereby transforming the ribose of the 2′-terminal nucleotide of 2-5A N-substituted morpholine. RNase L assays demonstrated that this novel 2-5A-antisense chimera had significant biological activity, thereby providing another potential tool for RNA ablation.
AB - A convergent synthetic approach was used to conjugate 2′,5′- oligoadenylate (2-5A, p5′A2′ [p5′A2′] np5′A) to phosphorodiamidate morpholino oligomers (morphants). To provide requisite quantities of 2-5A starting material, commercially and readily available synthons for solid-phase synthesis were adapted for larger scale solution synthesis. Thus, the tetranucleotide 5′- phosphoryladenylyl(2′→5′)adenylyl-(2′→5′) adenylyl(2′→5′)adenosine (p5′A2′p5′A2′] 2p5′A2′, tetramer 2-5A, 9) was synthesized starting with 2′,3′-O-dibenzoyl-N6,N6-dibenzoyl adenosine prepared from commercially available 5′-O-(4-monomethoxytrityl) adenosine. Coupling with N6-benzoyl-5′-O-(4,4′-dimethoxytrityl)- 3′-O-(tert-butyldimethylsilyl) adenosine-2′-(N,N-diisopropyl-2- cyanoethyl)phosphoramidite, followed by oxidization and deprotection, generated 5′-deprotected dimer 2-5A. Similar procedures lengthened the chain to form protected tetramer 2-5 A. The title product 9 p5′A(2′p5′A) 3 (tetramer 2-5A) was obtained through phosphorylation of the terminal 5′-hydroxy of the protected tetramer and removal of remaining protecting groups using concentrated ammonium hydroxide-ethanol (3:1, v/v) at 55°C and tetrabutylammonium fluoride (TBAF) in THF at room temperature, respectively. The 2-5A-phosphorodiamidate morpholino antisense chimera 11 (2-5A-morphant) was synthesized by covalently linking an aminolinker- functionalized phosphorodiamidate morpholino oligomer with periodate oxidized 2-5A tetramer (p5′A2′[p5′A2′]2p5′A). The resulting Schiff base was reduced with cyanoborohydride thereby transforming the ribose of the 2′-terminal nucleotide of 2-5A N-substituted morpholine. RNase L assays demonstrated that this novel 2-5A-antisense chimera had significant biological activity, thereby providing another potential tool for RNA ablation.
UR - http://www.scopus.com/inward/record.url?scp=15344345666&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=15344345666&partnerID=8YFLogxK
U2 - 10.1021/bc049778q
DO - 10.1021/bc049778q
M3 - Article
VL - 16
SP - 383
EP - 390
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
SN - 1043-1802
IS - 2
ER -