Pseudo-cyclic oligonucleotides: In vitro and in vivo properties

Zhiwei Jiang, Ekambar R. Kandimalla, Qiuyan Zhao, Ling X. Shen, Antonella Deluca, Nicola Normano, Mary Ruskowski, Sudhir Agrawal

Research output: Contribution to journalArticle

Abstract

We have designed and studied antisense oligodeoxynucleotides (oligonucleotides; oligos) which we call 'pseudo-cyclic oligonucleotides' (PCOs). PCOs contain two oligonucleotide segments attached through their 3'-3'- or 5'-5'-ends. One of the segments of the PCO is an antisense oligo complementary to a target mRNA, and the other is a short protective oligo that is 5-8 nucleotides long and complementary to the 3'- or 5'-end of the antisense oligo. As a result of complementarity between the antisense and protective oligo segments, PCOs form intramolecular pseudo-cyclic structures in the absence of the target RNA. The antisense oligo segment of PCOs used for the studies described here is complementary to an 18-nucleotide-long site on the mRNA of the protein kinase A regulatory subunit RIα (PKA-RIα). Thermal melting studies of PCOs in the absence and presence of the complementary RNA suggest that the pseudo-cyclic structures formed in the absence of the target RNA dissociate, bind to the target RNA, and form heteroduplexes. The results of RNase H cleavage assays suggest that PCOs bind to complementary RNA and activate RNase H in a manner similar to that of an 18-mer conventional antisense PS-oligo. In snake venom (a 3'-exonuclease) or spleen (a 5'-exonuclease) phosphodiesterase digestion studies, PCOs are more stable than conventional antisense oligos because of the presence of 3'-3'- or 5'-5'-linkages and the formation of intramolecular pseudo-cyclic structures. PCOs with a phosphorothioate antisense oligo segment inhibited cell growth of MDA-MB-468 and GEO cancer cell lines similar to that of the conventional antisense PS-oligo, suggesting efficient cellular uptake and target binding. The nuclease stability studies in mice suggest that PCOs have higher in vivo stability than antisense PS-oligos. The studies in mice showed similar pharmacokinetic and tissue distribution profiles for PCOs to those of antisense PS-oligos in general, but rapid elimination from selected tissues. Copyright (C) 1999 Elsevier Science Ltd.

Original languageEnglish
Pages (from-to)2727-2735
Number of pages9
JournalBioorganic and Medicinal Chemistry
Volume7
Issue number12
DOIs
Publication statusPublished - Dec 1999

Fingerprint

Oligonucleotides
Ribonuclease H
Complementary RNA
spleen exonuclease
Antisense Oligonucleotides
RNA
In Vitro Techniques
Nucleotides
Phosphodiesterase I
Messenger RNA
Snake Venoms
Tissue
Oligodeoxyribonucleotides
Phosphoric Diester Hydrolases
Tissue Distribution
Cyclic AMP-Dependent Protein Kinases
Pharmacokinetics
Freezing
Cell growth
Digestion

Keywords

  • Antisense
  • In vivo stability
  • Oligonucleotides
  • Pharmacokinetics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry
  • Drug Discovery
  • Pharmaceutical Science

Cite this

Pseudo-cyclic oligonucleotides : In vitro and in vivo properties. / Jiang, Zhiwei; Kandimalla, Ekambar R.; Zhao, Qiuyan; Shen, Ling X.; Deluca, Antonella; Normano, Nicola; Ruskowski, Mary; Agrawal, Sudhir.

In: Bioorganic and Medicinal Chemistry, Vol. 7, No. 12, 12.1999, p. 2727-2735.

Research output: Contribution to journalArticle

Jiang, Z, Kandimalla, ER, Zhao, Q, Shen, LX, Deluca, A, Normano, N, Ruskowski, M & Agrawal, S 1999, 'Pseudo-cyclic oligonucleotides: In vitro and in vivo properties', Bioorganic and Medicinal Chemistry, vol. 7, no. 12, pp. 2727-2735. https://doi.org/10.1016/S0968-0896(99)00217-5
Jiang, Zhiwei ; Kandimalla, Ekambar R. ; Zhao, Qiuyan ; Shen, Ling X. ; Deluca, Antonella ; Normano, Nicola ; Ruskowski, Mary ; Agrawal, Sudhir. / Pseudo-cyclic oligonucleotides : In vitro and in vivo properties. In: Bioorganic and Medicinal Chemistry. 1999 ; Vol. 7, No. 12. pp. 2727-2735.
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AU - Normano, Nicola

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AU - Agrawal, Sudhir

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