OLIGONUCLEOTIDES FOR REDUCTION OF PD-L1 EXPRESSION
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.