Conjugates of double-stranded oligonucleotides with poly(ethylene glycol) and keyhole limpet hemocyanin: a model for treating systemic lupus erythematosus.
Two types of oligonucleotides were synthesized with linker groups attached at the 5'-end. Both were repeating dimers of deoxyribocytidine and deoxyriboadenosine. A 20-mer was prepared with a thiol-containing linker, masked as a disulfide, and a 50-mer was prepared with a vicinal diol-containing linker. A tetraiodoacetylated poly(ethylene glycol) (PEG) derivative was synthesized and reacted with the thiol-containing 20-mer to provide an oligonucleotide PEG conjugate of precisely four oligonucleotides on each PEG carrier. The vicinal diol on the 50-mer was oxidized to an aldehyde and conjugated to keyhole limpet hemocyanin (KLH) to provide an oligonucleotide-KLH conjugate by reductive alkylation. The conjugates were annealed with complementary (TG)n strands. While the double-stranded oligonucleotide-KLH conjugate is an immunogen, eliciting the synthesis of antibodies against oligonucleotides, the PEG conjugate has the biological property of specifically suppressing (tolerizing) B cells which make antibodies against the immunizing oligonucleotide.
Jones
D S
DS
La Jolla Pharmaceutical Company, San Diego, California 92121.
Hachmann
J P
JP
Osgood
S A
SA
Hayag
M S
MS
Barstad
P A
PA
Iverson
G M
GM
Coutts
S M
SM
D000478:Alkylation; D000818:Animals; D000917:Antibody Formation; D000941:Antigens / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006433:Hemocyanins / Q000737:chemistry*; D007108:Immune Tolerance; D007114:Immunization; D017403:In Situ Hybridization; D008180:Lupus Erythematosus, Systemic / Q000628:therapy*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008810:Mice, Inbred C57BL; D009841:Oligonucleotides / Q000737:chemistry* / Q000276:immunology / Q000627:therapeutic use*; D011092:Polyethylene Glycols / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D000941:Antigens; D009841:Oligonucleotides; D011092:Polyethylene Glycols; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin