Mixed-backbone oligonucleotides as second-generation antisense agents with reduced phosphorothioate-related side effects

Bioorganic and Medicinal Chemistry Letters

Volumn 8, Issue 22, 1998, Pages 3269-3274

  Zhou, Wenqiang ^a   Agrawal, Sudhir ^a  

^a HYBRIDON INC   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ANTISENSE OLIGONUCLEOTIDE; NUCLEASE; PHOSPHOROTHIOIC ACID DERIVATIVE; RIBONUCLEASE III;

ANIMAL TISSUE; ARTICLE; DRUG DISPOSITION; DRUG MECHANISM; DRUG STRUCTURE; ENZYME ACTIVITY; HUMAN; HUMAN TISSUE; MOUSE; NONHUMAN; PARTIAL THROMBOPLASTIN TIME; STRUCTURE ANALYSIS; THERMOSTABILITY; TISSUE DISTRIBUTION;

ANIMALIA;

EID: 0032541985     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0960-894X(98)00591-5     Document Type: Article

References (25)

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17. 31PNMR, and MALDI-TOF MS.
18. 2 (15 mM). To each solution, 0.004 units of SVPD from crotallus durissus (Boehringer Mannheim) was added. The reaction was carried out for 24 h. at 37 °C.
19. 13. In a parallel study, it was found that substitution of the phosphorothioate linkage with a phosphodiester linkage in the PS-oligo (oligo 1, Table 1) reduced the modified oligos' stability towards SVPD (data not shown).
20. 14. Tm were recorded using a GBC 920 Spectrophotometer (GBC Scientific Equipment, Victoria, Australia). Oligos were mixed with complementary RNA phosphodiester (30-mer, 5′-ACCGCCGCCAGUGAGGA GGCACGCAGCCUU-3′) in a buffer containing 10 mM Pipes, 1 mM EDTA, and 100 mM NaCl. The Tm values were obtained from the first derivative plots.
21. 2, 100 mM KCl, 1 mM DTT, 200 mM Tris (pH 7.5), and 5% glycerol. After annealing, 0.078 unit of RNase H (Pharmacia) was added to each solution. The mixture were then incubated at 37 °C for 10 min. The reactions were then quenched by adding 20 μL of gel loading dye to each reaction mixture. The resultant samples were analyzed by 20% PAGE and subjected to autoradiography.
22. 16. General procedure for aPTT: Plasma was obtained from citrated human blood. Serial dilution of the oligos in 0.9% NaCl UPS (saline) were provided in final cones, of 6.25, 12.5, 25, 50 and 100 μg/mL of oligo in plasma. After addition of the oligo samples, the plasma was incubated at 37 °C for 15 min, with gentle agitation. Plasma exposed to vehicle in the same ratio (v/v) as the oligos, and untreated plasma served as negative controls. The assay was conducted in duplicate, providing at least 2 replication for each tube.
23. 17. Kandimalla, E.; Shaw, D.; Agrawal, S., Bioorg. Med. Chem. Lett. 1998, 8, 2103.
24. 18. Oligo 1 and 6 (1 mg) were administrated intravenously in mice (female, CD-1, 20 -22 g) through the tail vein. Blood samples (200 μL) was collected at 1 h post-dosing. The oligos were extracted from the plasma by the same procedure as ref. 19, and analyzed for their stability by CGE.
25. 19. Grindel, J.; Musick, T.; Jiang, Z.; Roskey, A.; Agrawal, S. Antisense Res. Dev. 1998, 8, 43.