Phosphorothioate oligodeoxynucleotides

Stein, C. A. and Cohen, J. S., Phosphorothioate oligodeoxynucleotide analogues, in Oligodeoxynucleotides — Antisense Inhibitors of Gene Expression, Cohen, J. S., Ed., CRC Press, Boca Raton, FL, 1989, 97. 

Previous studies have reported that ERKs are characteristically associated with cell proliferation and protection from apoptosis (Bl, XI), while activation of JNK and p38 MAPK can promote apoptosis in many systems, including B lymphocytes (G5), cerebellar granule cells (K3), hematopoietic cells (K8), and neuronal cells (M3, XI). On the other hand, a recent report found that a pyridinyl imidazole, SB 202190, the specific inhibitor of p38 MAPK, by itself was sufficient to induce apoptosis in T lymphocyte Jurkat cells (N2). Moreover, Th-2-derived cytokine IL-5, the ERK activator and antiapoptotic factor for eosinophils, could also activate p38 MAPK in human eosinophils (BIO). We recently reported that cytokine IL-3, IL-5, and GM-CSF could prolong survival of human eosinophilic leukemic (EoL-1) cells through the transient activation of ERK (W15). On the other hand, activation of p38 MAPK in EoL-1 cells by the NSAID sodium salicylate (NaSal) could lead to apoptosis (W15). We also found that the suppression of ERK using ERK antisense phosphorothioate oligodeoxynucleotides could promote the apoptosis of peripheral blood eosinophils (W16). Moreover, we found that dexamethasone-induced apoptosis and activation of JNK and p38 MAPK activity in eosinophils are regulated by caspases (Z2). 

Phillips, J.A., Craig, S.J., Bayley, D., Christian, R.A., Geary, R. and Nicklin, P.L. (1997) Pharmacokinetics, metabolism, and elimination of a 20-mer phosphorothioate oligodeoxynucleotide (CGP 69846A) after intravenous and subcutaneous administration. Biochem. Pharmacol, 54, 657-668. 

Wu, D., R. Boado, and W. Pardridge. 1996. Pharmacokinetics and blood-brain barrier transport of [3H]-biotinylated phosphorothioate oligodeoxynucleotide conjugated to a vector-mediated drug delivery system. J Pharmacol Exp Ther 276 206. 

Table 4.1 Antisense phosphorothioate oligodeoxynucleotides studied in human clinical trials.

D.K. Monteith, and A.A. Levin. 1997. Antisense oligonucleotide inhibitors for the treatment of cancer 1. Pharmacokinetic properties of phosphorothioate oligodeoxynucleotides. Anti-Cancer Drug Design 12 383-393. 

E. Chueng, and A.A. Levin. 2002. Development of an ultrasensitive noncompetitive hybridization-ligation enzyme-linked immunosorbent assay for the determination of phosphorothioate oligodeoxynucleotide in plasma. Anal. Biochem. 304 19-25. 

In animal studies, there is overwhelming evidence of biological activity of ASOs upon IV injection of simple solutions. Given the excellent solution stability and solubility possessed by phosphorothioate oligodeoxynucleotides, it has been rela-... 

Relative to the unmodified phosphorothioate oligodeoxynucleotides, these data demonstrate the increasing benefit of MOE substituents present as either a hemi-mer, with MOEs at the 3 -termini only or a gap-mer, with MOEs present at both the 3 - and 5 -termini and lastly, a full MOE substitution at every sugar location. These MOE gap-mer chemistries are demonstrating clinical efficacy by the subcutaneous... 

Stein CA, Tokinson JL, Yakubov L, Phosphorothioate oligodeoxynucleotides antisense inhibitors of gene expression Pharmacol Ther 1991 52 365-384,... 

Butler M, Stecker K, Bennett CF. Cellular distribution of phosphorothioate oligodeoxynucleotides in normal rodent tissues. Lab Invest 1997 77(4) 379-88. 

Henry SP, Monteith DK, Matson JE, Mathison BH, Loveday KS, Winegar RA, Lee PS, Riccio ES, Bakke JP, Levin AA. Assessment of the genotoxic potential of ISIS 2302 a phosphorothioate oligodeoxynucleotide. Mutagenesis 2002 17(3) 201-9. 

Henry SP, Beattie G, Yeh G, Chappel A, Giclas PC, Mortari A, Jagels MA, Kornbrust DJ, Levin AA. Complement activation is responsible for acute toxicities in rhesus monkeys treated with a phosphorothioate oligodeoxynucleotide. Inti J Immunopharmacol 2002 2(12) 1657-66. 

Liang H, Nishioka Y, Reich CF, Pisetsky DS, Lipsky PE. Activation of human B cells by phosphorothioate oligodeoxynucleotides. / Clin Invest 1996 98(5) 1119-29. 

Branda RF, Moore AL, Lafayette AR, Mathews L, Hong R, Zon G, Brown T, McCormack JJ. Amplification of antibody production by phosphorothioate oligodeoxynucleotides. I Lab Clin Med 1996 128(3) 329-38. 

In a more recent study in which the level of intact drug was carefully evaluated using capillary gel electrophoresis, the pharmacokinetics of ISIS 2302, a 20-mer phosphorothioate oligodeoxynucleotide, after a 2-h infusion, were determined. Doses from 0.06 to 2.0 mgkg were studied and the peak plasma concentrations were shown to increase linearly with dose, with the 2 mg/kg dose resulting in peak plasma concentrations of intact drug of about 9.5 Ltg/mL. Clearance from plasma, however, was dose dependent, with the 2 mg/kg dose having a clearance of 1.28 mL min kg-, whereas that of 0.5 mg/kg was... 

ISIS 5132. ISIS 5231 is a phosphorothioate oligodeoxynucleotide designed to inhibit C-raf kinase. In phase I studies, this drug vvas shown to reduce C-raf kinase levels in peripheral blood cells and displayed activity in patients with ovarian cancer (251). It too is completing phase II evaluation (for review, see Ref 247). 

In Vitro. In our laboratory, we have evaluated the toxicities of scores of phosphorothioate oligodeoxynucleotides in a significant number of cell lines in tissue culture. As a general rule, no significant cytotoxicity is induced at concentrations below 100 jllM oligonucleotide. Additionally, with a few exceptions, no significant effect on macromolecular synthesis is observed at concentrations below 100(188,192). 

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