Stabilized immunomodulatory oligonucleotides

摘要

The invention provides immunostimulatory oligonucleotides having at least one CpG dinucleotide and a secondary structure at the 5′- or 3′-end. These oligonucleotides have either reduced or improved immunostimulatory properties. The invention establishes that 5′-terminal secondary structures affect immunostimulatory activity significantly more than those at the 3′-end. The invention also provides methods for increasing or decreasing the immunostimulatory activity of a CpG-containing nucleic acid.

主权项

1. A method for treating cancer, infection, inflammatory disease or autoimmune disease comprising administering to a patient an immunostimulatory nucleic acid, having secondary structure formed by intermolecular hydrogen bonding between two oligonucleotide compounds, wherein each of the oligonucleotide compounds comprise the general structure of:
Domain A-Domain B-Domain C,  (I)wherein Domain A is 5′-3′ DNA not having a palindromic or self-complementary domain and containing at least one dinucleotide selected from the group consisting of CpG, C*pG, C*pG* and CpG*, wherein C is cytidine or 2′-deoxycitidine, G is guanosine or 2′-deoxyguanosine, C* is 2′-deoxythymidine, 1-(2′-deoxy-β-D-ribofuranosyl)-2-oxo-7-deaza-8-methyl-purine, 2′ dideoxy-5-halocytosine, 2′-dideoxy-5-nitrocytosine, arabinocytidine, 2′-deoxy-2′-substitutedarabinocytidine, 2′-O-substituted arabinocytidine, 2′-deoxy-5-hydroxycytidine, 2′-deoxy-N4-alkyl-cytidine, 2′-deoxy-4-thiouridine, or other non-natural pyrimidine nucleosides, G*is 2′ deoxy-7-deazaguanosine, 2′-deoxy-6-thioguanosine, arabinoguanosine, 2′-deoxy-2′-substituted-arabinoguanosine, 2′-O-substituted-arabinoguanosine, 2′-deoxyinosine, or other non-natural purine nucleoside, and p is an internucleoside linkage selected from the group consisting of phosphodiester, phosphorothioate, and phosphorodithioate, wherein Domain B is a non-nucleoside linker joining Domain A and Domain C, wherein Domain C is 3′-5′ DNA or RNA having a palindromic or self-complementary domain allowing for intermolecular hydrogen bonding, and which can or cannot have a dinucleotide selected from the group consisting of CpG, C*pG, C*pG* and CpG*, wherein C is cytidine or 2′deoxycitidine, G is guanosine or 2′ deoxyguanosine, C* is 2′-deoxythymidine, 1-(2′-deoxy-β-D-ribofuranosyl)-2-oxo-7-deaza-8-methyl-purine, 2′-dideoxy-5-halocytosine, 2′-dideoxy-5-nitrocytosine, arabinocytidine, 2′-deoxy-2′-substituted arabinocytidine, 2′-O-substituted arabinocytidine, 2′-deoxy-5-hydroxycytidine, 2′-deoxy-N4-alkyl-cytidine, 2′-deoxy-4-thiouridine, or other non-natural pyrimidine nucleosides, G* is 2′ deoxy-7-deazaguanosine, 2′-deoxy-6-thioguanosine, arabinoguanosine, 2′-deoxy-2′substituted-arabinoguanosine, 2′-O-substituted-arabinoguanosine, 2′-deoxyinosine, or other non-natural purine nucleoside, p is an internucleoside linkage selected from the group consisting of phosphodiester and phosphorothioate, and wherein each oligonucleotide compound comprises from about 12 to about 50 nucleotides in length.