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

被引:59
作者
JONES, DS
BARSTAD, PA
FEILD, MJ
HACHMANN, JP
HAYAG, MS
HILL, KW
IVERSON, GM
LIVINGSTON, DA
PALANKI, MS
TIBBETTS, AR
YU, L
COUTTS, SM
机构
[1] La Jolla Pharmaceutical Company, San Diego, California 92121
关键词
D O I
10.1021/jm00012a013
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
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 tetrabro-moacetylated platform by displacement with sulfhydryl-terminated linkers. The tetrabro-moacetylated 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.
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收藏
页码:2138 / 2144
页数:7
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