Immunospecific Reduction of Antioligonucleotide Antibody-Forming Cells with a Tetrakis-oligonucleotide Conjugate (LJP 394), a Therapeutic Candidate for the Treatment of Lupus Nephritis

Abstract

A discrete tetravalent conjugate, 7a (LJP 394), consisting of four oligonucleotides attached to a common carrier or platform was prepared. Single-stranded oligonucleotide 20-mers consisting of alternating deoxycytidine-deoxyadenosine nucleotides, (CA)10, were attached to a tetrabromoacetylated platform by displacement with sulfhydryl-terminated linkers. The tetrabromoacetylated platform 3a was synthesized in three steps using triethylene glycol bis-(chloroformate). The single-stranded conjugate was characterized by polyacrylamide gel electrophoresis, DNA sequencing, phosphate analysis, carbon and nitrogen combustion analysis, and correlation of stoichiometry to conversion in the conjugation process. HPLC and capillary electrophoretic methods were developed to evaluate purity. The tetrakis, single-stranded conjugate was annealed with a stoichiometric amount of a complementary single-stranded oligonucleotide 20-mer consisting of alternating thymidine-deoxyguanosine nucleotides, (TG)10. The double-stranded conjugate LJP 394 was characterized by melt temperature and hyperchromicity, phosphate analysis, and carbon and nitrogen combustion analysis. LJP 394 inhibits binding of DNA to anti-double-stranded oligonucleotide antibodies and reduces anti-oligonucleotide-specific plaque (antibody)-forming cells in an immunized mouse model by a proposed mechanism involving cross-linking B cell surface immunoglobins.

Impact of DNA Hairpin Folding Energetics on Antibody–ssDNA Association

“…Because the binding of anti-DNA Abs to either DNA or ligands that mimic DNA is common to all three pathways, disruption of these interactions has been proposed as a novel drug design strategy. 3,8,[12][13][14][15] Thus, structural and thermodynamic analysis of Ab-DNA recognition is of interest.…”

Evidence for Structural Plasticity of Heavy Chain Complementarity-determining Region 3 in Antibody–ssDNA Recognition

“…3,[12][13][14] Anti-DNA Abs represent a challenging ligand design target, however, due to their inherent cross-reactivity. Several studies have reported the binding of non-DNA ligands to 0022-2836/$ -see front matter q 2005 Elsevier Ltd. All rights reserved.…”

New and emerging treatments for lupus nephritis

“…Its mechanism of action is to induce tolerance by cross-linking anti-dsDNA surface antibodies on B-cells, leading to B-cell anergy or apoptosis and reduced anti-dsDNA production. 45 In a phase III trial intended to evaluate whether abetimus could decrease the risk of nephritic fl ares, 317 patients with a history of lupus nephritis and elevated anti-dsDNA antibodies were randomized to abetimus or placebo. 46 Abetimus did not prolong time to renal fl are, time to major SLE fl are, or time to initiation of standard induction therapy.…”