Activation of the alternative pathway of complement by a phosphorothioate oligonucleotide: potential mechanism of action.
Intravenous infusion of high doses of phosphorothioate oligonucleotides in monkeys has been associated with transient alterations in hematologic and hemodynamic parameters, which appear to be secondary to complement activation. ISIS 2302, a phosphorothioate oligonucleotide specific for human intracellular adhesion molecule-1, was used to further characterize complement activation in monkeys. Complement activation occurred selectively through the alternative pathway resulting in increased plasma concentrations of the complement split products Bb, C3a and C5a. Marked fluctuations in circulating neutrophil counts and reductions in cardiac output were closely associated with peak production of anaphylatoxins C3a and C5a. Changing both dose and infusion duration revealed that complement activation is related to plasma levels of oligonucleotide, and that there is a minimum threshold concentration of approximately 50 micrograms/ml of ISIS 2302 that is required to activate complement. Dose regimens in which plasma concentrations do not exceed this threshold do not result in complement activation. Further investigation reveals that plasma concentrations of a key regulatory component of the alternative pathway, Factor H, were also decreased after administration of ISIS 2302. Decreases in Factor H levels are suggestive of a possible mechanism of complement activation. Direct interaction between ISIS 2302 and Factor H was demonstrated in a competition assay, where increasing concentrations of ISIS 2302 eluted Factor H from a heparin-sepharose column. These data demonstrate a clear correlation between plasma oligonucleotide concentrations and complement activation. Interactions between ISIS 2302 and Factor H may lead to activation of the alternative complement pathway.
Henry
S P
SP
Department of Toxicology, Isis Pharmaceuticals, Inc. Carlsbad, California, USA.
Giclas
P C
PC
Leeds
J
J
Pangburn
M
M
Auletta
C
C
Levin
A A
AA
Kornbrust
D J
DJ
D000818:Animals; D003170:Complement Pathway, Alternative / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005260:Female; D008252:Macaca fascicularis; D020319:Oligodeoxyribonucleotides, Antisense; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D054735:Phosphorothioate Oligonucleotides; D013873:Thionucleotides / Q000494:pharmacology*
D020319:Oligodeoxyribonucleotides, Antisense; D016376:Oligonucleotides, Antisense; D009943:Organophosphorus Compounds; D054735:Phosphorothioate Oligonucleotides; D013873:Thionucleotides; C089753:alicaforsen