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

Jones, David S.; Barstad, Paul A.; Feild, Mark J.; Hachmann, John P.; Hayag, Merle S.; Hill, Kenneth W.; Iverson, G. Michael; Livingston, Douglas A.; Palanki, Moorthy S.; et al.

La Jolla Pharmaceutical Company, 6455 Nancy Ridge Drive, San Diego, CA 92121

A discrete tetravalent conjugate, 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 was synthesized in three steps using triethylene glycol bis(chloroformate). The single-stranded conjugate was characterized by PAGE, DNA sequencing, phosphate anal., carbon and nitrogen combustion anal., and correlated 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 amt. 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 temp. and hyperchromicity, phosphate anal., and carbon and nitrogen combustion anal. 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 crosslinking B cell surface immunoglobins.

Published in the Journal of Medicinal Chemistry, 1995, 38.