IMMUNOSPECIFIC REDUCTION OF ANTIOLIGONUCLEOTIDE ANTIBODY-FORMING-CELLS WITH A TETRAKIS-OLIGONUCLEOTIDE CONJUGATE (LJP-394), A THERAPEUTIC CANDIDATE FOR THE TREATMENT OF LUPUS NEPHRITIS

A discrete tetravalent conjugate, 7a (LJP 394), consisting of four oli gonucleotides attached to a common carrier or platform was prepared. S ingle-stranded oligonucleotide 20-mers consisting of alternating deoxy cytidine-deoxyadenosine nucleotides, (CA)(10), were attached to a tetr abro-moacetylated platform by displacement with sulfhydryl-terminated linkers. The tetrabro-moacetylated platform 3a was synthesized in thre e steps using triethylene glycol bis-(chloroformate). The single-stran ded conjugate was characterized by polyacrylamide gel electrophoresis, DNA sequencing, phosphate analysis, carbon and nitrogen combustion an alysis, and correlation of stoichiometry to conversion in the conjugat ion process. HPLC and capillary electrophoretic methods were developed to evaluate purity. The tetrakis, single-stranded conjugate was annea led with a stoichiometric amount of a complementary single-stranded ol igonucleotide 20-mer consisting of alternating thymidine-deoxyguanosin e nucleotides, (TG)(10). The double-stranded conjugate LJP 394 was cha racterized by melt temperature and hyperchromicity, phosphate analysis , and carbon and nitrogen combustion analysis. LJP 394 inhibits bindin g of DNA to anti-double-stranded oligonucleotide antibodies and reduce s anti-oligonucleotide-specific plaque (antibody)-forming cells in an immunized mouse model by a proposed mechanism involving cross-linking B cell surface immunoglobins.