JPWO2020069044A5 - - Google Patents

Claims (38)

A modified antisense oligonucleotide (AON) comprising at least one 2'-deoxy-2'-fluoro-β-D-arabinonucleotide (2'-FANA modified nucleotide), wherein the AON binds to Foxp3 mRNA. 2. The AON of claim 1, which is a hybrid chimeric AON comprising at least one 2'-FANA modified nucleotide and at least one unmodified deoxyribonucleotide, said AON being a 2'-FANA AON. 3. The AON of claim 2, wherein said 2'-FANA modified nucleotides are arranged according to any of Formulas 1-16. 4. The AON of claim 3, wherein said 2'-FANA modified nucleotides are arranged according to Formula 6. The internucleotide linkages between the nucleotides of the 2'-FANA modified nucleotides are phosphodiester bonds, phosphotriester bonds, phosphorothioate bonds (5'OP(S)O-3O-, 5'SP(O) O-3'-O-, and 5'OP(O)O-3'S-), phosphorodithioate bond, Rp-phosphorothioate bond, Sp-phosphorothioate bond, boranophosphate bond, methylene bond (methylimino ), amide bond (3′-CH ₂ -CO-NH-5′ and 3′-CH ₂ -NH-CO-5′), methylphosphonate bond, 3′-thioformacetal bond, (3′S-CH ₂ -O5'), an amide bond (3'CH2 _- C(O)NH-5'), a phosphoramidate group, and combinations thereof. The 2′-FANA AON is
about 0 to about 20 2'-deoxy-2'-fluoro-β-D-arabinonucleotides at the 5' end and 3 flanking a sequence comprising about 0 to about 20 deoxyribonucleotide residues; 3. The AON of claim 2, comprising from about 0 to about 20 2'-deoxy-2'-fluoro-β-D-arabinonucleotides at the 'end. The 2'-FANA AON has SEQ ID NO: 1 to 9, SEQ ID NO: 11 to 19, SEQ ID NO: 21 to 29, SEQ ID NO: 31 to 138, SEQ ID NO: 139 to 192, SEQ ID NO: at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16 of 193-302, or sequences complementary thereto , at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 consecutive nucleotides. A pharmaceutical composition comprising a modified antisense oligonucleotide (AON) comprising at least one 2'-deoxy-2'-fluoro-β-D-arabinonucleotide (2'-FANA modified nucleotide) and a pharmaceutically acceptable carrier A pharmaceutical composition, wherein said AON binds to Foxp3 mRNA. 9. The composition of claim 8, wherein said AON is a hybrid chimeric AON comprising at least one 2'-FANA modified nucleotide and at least one unmodified deoxyribonucleotide, and said AON is a 2'-FANA AON. The 2′-FANA AON is
about 0 to about 20 2'-deoxy-2'-fluoro-β-D-arabinonucleotides at the 5' end and 3 flanking a sequence comprising about 0 to about 20 deoxyribonucleotide residues; 10. The composition of claim 9, comprising from about 0 to about 20 2'-deoxy-2'-fluoro-β-D-arabinonucleotides at the 'end. The 2'-FANA AON has SEQ ID NO: 1 to 9, SEQ ID NO: 11 to 19, SEQ ID NO: 21 to 29, SEQ ID NO: 31 to 138, SEQ ID NO: 139 to 192, SEQ ID NO: at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16 of 193-302, or sequences complementary thereto , at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 consecutive nucleotides. A pharmaceutical composition for reducing the expression level of the Foxp3 gene in a cell, comprising:
comprising at least one antisense oligonucleotide (AON) ;
said AON binds to Foxp3 mRNA, and said AON comprises at least one 2'-deoxy-2'-fluoro-β-D-arabinonucleotide (2'-FANA modified nucleotide);
A pharmaceutical composition , thereby reducing the expression level of Foxp3. 13. The composition of claim 12, wherein said cells are regulatory T cells (Treg). 14. The composition of claim 13, wherein said Tregs express the cell markers CD4 and CD25. 13. The composition of claim 12, wherein said AON is a hybrid chimeric AON comprising at least one 2'-FANA modified nucleotide and at least one unmodified deoxyribonucleotide, and said AON is a 2'-FANA AON. The 2'-FANA AON has SEQ ID NO: 1 to 9, SEQ ID NO: 11 to 19, SEQ ID NO: 21 to 29, SEQ ID NO: 31 to 138, SEQ ID NO: 139 to 192, SEQ ID NO: at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16 of 193-302, or sequences complementary thereto , at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 consecutive nucleotides . A pharmaceutical composition for increasing anti-tumor immunity in a subject in need thereof, comprising:
comprising at least one antisense oligonucleotide ( AON );
said AON binds to Foxp3 mRNA, and said AON comprises at least one 2'-deoxy-2'-fluoro-β-D-arabinonucleotide (2'-FANA modified nucleotide);
A pharmaceutical composition thereby increasing anti-tumor immunity. 18. The composition of claim 17, wherein said AON reduces the activity of regulatory T cells (Treg). 19. The composition of claim 18, wherein said Tregs express the cell markers CD4 and CD25. 18. The composition of claim 17, wherein said AON induces Treg apoptosis. 18. The composition of claim 17, wherein said AON increases the activity of immune cells, thereby increasing anti-tumor immunity. 22. The composition of claim 21, wherein said immune cells are CD8 ⁺ T cells, CD4 ⁺ T cells, B cells, natural killer cells, macrophages, dendritic cells, or combinations thereof. 18. The composition of claim 17, wherein said AON is a hybrid chimeric AON comprising at least one 2'-FANA modified nucleotide and at least one unmodified deoxyribonucleotide, and said AON is a 2'-FANA AON. The 2'-FANA AON has SEQ ID NO: 1 to 9, SEQ ID NO: 11 to 19, SEQ ID NO: 21 to 29, SEQ ID NO: 31 to 138, SEQ ID NO: 139 to 192, SEQ ID NO: at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16 of 193-302, or sequences complementary thereto , at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 consecutive nucleotides . A pharmaceutical composition for treating cancer in a subject in need thereof, comprising:
comprising at least one antisense oligonucleotide ( AON );
said AON binds to Foxp3 mRNA, and said AON comprises at least one 2'-deoxy-2'-fluoro-β-D-arabinonucleotide (2'-FANA modified nucleotide);
A pharmaceutical composition thereby treating said cancer. 26. The composition of claim 25, wherein said AON reduces the expression level of the Foxp3 gene. 26. The composition of claim 25, wherein said AON is a hybrid chimeric AON comprising at least one 2'-FANA modified nucleotide and at least one unmodified deoxyribonucleotide, and said AON is a 2'-FANA AON. The 2'-FANA AON has SEQ ID NO: 1 to 9, SEQ ID NO: 11 to 19, SEQ ID NO: 21 to 29, SEQ ID NO: 31 to 138, SEQ ID NO: 139 to 192, SEQ ID NO: at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16 of 193-302, or sequences complementary thereto , at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 consecutive nucleotides . 28. The composition of claim 27, wherein said 2'-FANA AON increases anti-tumor immunity in said subject. 28. The composition of claim 27, wherein said 2'-FANA AON decreases regulatory T cell (Treg) activity and/or increases immune cell activity. 26. The composition of Claim 25, further comprising a pharmaceutically acceptable carrier. 26. The composition of claim 25, administered in combination with an immunotherapeutic agent and/or a chemotherapeutic agent. The immunotherapeutic agent and/or chemotherapeutic agent is a checkpoint inhibitor, vaccine, chimeric antigen receptor (CAR)-T cell therapy, anti-PD-1 antibody (nivolumab or pembrolizumab), anti-PD-L1 antibody (atezolizumab, 33. The composition of claim 32 selected from the group consisting of avelumab, or durvalumab), and combinations thereof . 33. The composition of claim 32, wherein said immunotherapeutic and/or chemotherapeutic agent is administered prior to, concurrently with, or after administration of said AON. 26. The composition of claim 25, administered in combination with radiation therapy. 36. The composition of claim 35, wherein said radiotherapy is administered prior to, concurrently with, or after administration of said AON . 26. The composition of claim 25, wherein said cancer is selected from the group consisting of breast cancer, liver cancer, ovarian cancer, pancreatic cancer, lung cancer, melanoma, and glioblastoma. 38. The composition of claim 37, wherein said cancer is lung cancer.