Oligonucleotides for reduction of pd-l1 expression

Abstract

The present invention relates to antisense oligonucleotides that are capable of reducing expression of PD-L1 in a target cell. The oligonucleotides hybridize to PD-L1 mRNA. The present invention further relates to conjugates of the oligonucleotide and pharmaceutical compositions and methods for treatment of viral liver infections such as HBV, HCV and HDV; parasite infections such as malaria, toxoplasmosis, leishmaniasis and trypanosomiasis or liver cancer or metastases in the liver using the oligonucleotide.

1 . An antisense oligonucleotide conjugate comprising:
 a. an oligonucleotide (Region A) comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length with at least 90% complementarity to a PD-L1 target nucleic acid; and   b. at least one asialoglycoprotein receptor targeting conjugate moiety (Region C) covalently attached to the oligonucleotide in a).   
     
     
         2 . The oligonucleotide conjugate of  claim 1 , wherein the contiguous nucleotide sequence is complementary to a target nucleic acid selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3. 
     
     
         3 . The oligonucleotide conjugate of  claim 1 , wherein the contiguous nucleotide sequence is complementary to a sub-sequence of the target nucleic acid, wherein the subsequence is selected from the group consisting of position 371-3068, 5467-12107 and 15317-19511 on SEQ ID NO: 1 
     
     
         4 . The oligonucleotide conjugate of  claim 1 , wherein the contiguous nucleotide sequence is complementary to a sub-sequence of the target nucleic acid, wherein the subsequence is selected from the group consisting of A221, A360, A180, A160 and A269. 
     
     
         5 . The oligonucleotide conjugate of  claim 1  wherein the oligonucleotide comprises a sequence selected from SEQ ID NO: 466, 640, 342, 287 and 566. 
     
     
         6 . The oligonucleotide conjugate of  claim 1 , wherein the contiguous nucleotide sequence comprises one or more modified nucleosides, such as one or more 2′ sugar modified nucleosides. 
     
     
         7 . The oligonucleotide conjugate of  claim 6 , wherein the one or more 2′ sugar modified nucleoside is independently selected from the group consisting of 2′-O-alkyl-RNA, 2′-O-methyl-RNA, 2′-alkoxy-RNA, 2′-O-methoxyethyl-RNA, 2′-amino-DNA, 2′-fluoro-DNA, arabino nucleic acid (ANA), 2′-fluoro-ANA and LNA nucleosides. 
     
     
         8 . The oligonucleotide conjugate of  claim 6 , wherein the all the modified nucleosides are LNA nucleosides. 
     
     
         9 . The oligonucleotide conjugate of  claim 7 , wherein the contiguous nucleotide sequence comprises at least one modified internucleoside linkage, such as at least one phosphorothioate internucleoside linkage. 
     
     
         10 . The oligonucleotide conjugate of  claim 1 , wherein the oligonucleotide is a gapmer. 
     
     
         11 . The oligonucleotide conjugate of  claim 10 , wherein the gapmer has formula 5′-D′-F-G-F′-3′ or 5′-F-G-F′-D″-3′, where region F and F′ independently comprise 1-7 modified nucleosides, G is a region between 6 and 16 nucleosides which are capable of recruiting RNaseH and region D′ or D″ is optional and comprise 0-5 phosphodiester linked nucleosides. 
     
     
         12 . The oligonucleotide conjugate of  claim 1 , wherein the asialoglycoprotein receptor targeting conjugate moiety comprises at least one carbohydrate moiety selected from group consisting of galactose, galactosamine, N-formyl-galactosamine, N-acetylgalactosamine, N-propionyl-galactosamine, N-n-butanoyl-galactosamine and N-isobutanoylgalactosamine. 
     
     
         13 . The oligonucleotide conjugate of  claim 1 , wherein the asialoglycoprotein receptor targeting conjugate moiety is mono-valent, di-valent, tri-valent or tetra-valent. 
     
     
         14 . The oligonucleotide conjugate of  claim 1 , wherein the asialoglycoprotein receptor targeting conjugate moiety is a tri-valent N-acetylgalactosamine (GalNAc) moiety. 
     
     
         15 . The oligonucleotide conjugate of  claim 1 , wherein the asialoglycoprotein receptor targeting conjugate moiety is the trivalent GalNAc moiety in  FIG. 3 . 
     
     
         16 . The oligonucleotide conjugate of  claim 1 , wherein the oligonucleotide conjugate is selected from CMP ID NO: 766_2, 767_2, 768_2, 769_2 and 770_2. 
     
     
         17 . An antisense oligonucleotide comprising an oligonucleotide or a contiguous nucleotide sequence according to  claim 1 . 
     
     
         18 . A pharmaceutical composition comprising the oligonucleotide conjugate  claim 1  and a pharmaceutically acceptable diluent, solvent, carrier, salt and/or adjuvant. 
     
     
         19 . An in vivo or in vitro method for modulating PD-L1 expression in a target cell which is expressing PD-L1, said method comprising administering an oligonucleotide conjugate of  claim 1  in an effective amount to said cell. 
     
     
         20 . A method for treating a disease comprising administering a therapeutically or prophylactically effective amount of an oligonucleotide conjugate of  claim 1  to a subject suffering from or susceptible to the disease. 
     
     
         21 . The method of  claim 20 , wherein the disease is selected from viral liver infections such as HBV, HCV and HDV; parasite infections such as malaria, toxoplasmosis, leishmaniasis and trypanosomiasis; and liver cancer or metastases in the liver. 
     
     
         22 . A method for restoration of immune response against a virus or parasite comprising administering a therapeutically or prophylactically effective amount of an oligonucleotide conjugate of  claim 1  to a subject infected with a virus or parasite. 
     
     
         23 . The method of  claim 22 , wherein the restoration of the immune response is an increase in the liver of CD8+ T cells specific to one or more HBV antigens when compared to a control.