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JP-2026075623-A - Oligonucleotides for reducing PD-L1 expression

JP2026075623AJP 2026075623 AJP2026075623 AJP 2026075623AJP-2026075623-A

Abstract

[Problem] The present invention relates to an antisense oligonucleotide that can reduce the expression of PD-L1 in target cells. [Solution] Oligonucleotides hybridize to PD-L1 mRNA. The present invention further relates to oligonucleotide conjugates and pharmaceutical compositions, as well as methods for treating viral hepatitis infections such as HBV, HCV, and HDV; parasitic infections such as malaria, toxoplasmosis, leishmaniasis, and trypanosomiasis; and liver cancer or metastasis in the liver using oligonucleotides. [Selection Diagram] None

Inventors

  • ルゲ ピーダスン
  • ハッサン ジャバンバクト
  • マリーネ ジャクエロット
  • セーアン オトスン
  • スーファローヌ ルワンセイ

Assignees

  • エフ． ホフマン－ラ ロシュ アーゲー

Dates

Publication Date
20260508
Application Date
20251211
Priority Date
20160314

Claims (20)

  1. An antisense oligonucleotide comprising a continuous nucleotide sequence of 10 to 30 nucleotides in length, capable of reducing PD-L1 expression, wherein the continuous nucleotide sequence is at least 90% complementary to the PD-L1 target nucleic acid, and the continuous nucleotide sequence is complementary to a partial sequence of the target nucleic acid, wherein the partial sequence is selected from the group consisting of positions 5467-12107, 15317-15720, 15317-18083, 15317-19511, 9812-9859, 15690-15735, 371-3068, 7300-7333, 8028-8072, 11787-11873, and 18881-19494 of SEQ ID NO: 1.
  2. It has at least 90% complementarity to the PD-L1 target nucleic acid, and the sequence numbers are 466, 640, 6, 8, 9, 13, 41, 42, 58, 77, 92, 111, 128, 151, 164, 166, 169, 171, 222, 233, 245, 246, 250, 251, 252, 256, 272, 273, 287, 292, 303, 314, 318, 320, 324, 336, 342, 343, 344, 345, 346, 349, 359, 360, 374, 408, 409, 415, 417, 424, 429, 430, 458, 464, 4 An antisense oligonucleotide comprising a sequence of 10 to 30 nucleotides in length, having at least 90% identity with a sequence selected from the sequences 74, 490, 493, 512, 519, 529, 533, 534, 547, 566, 567, 578, 582, 601, 619, 620, 636, 637, 638, 645, 650, 651, 652, 653, 658, 659, 660, 665, 678, 679, 680, 682, 683, 684, 687, 694, 706, 716, 728, 733, 734, and 735.
  3. The aforementioned continuous nucleotide sequence is sequence numbers 6, 8, 9, 13, 41, 42, 58, 77, 92, 111, 128, 151, 164, 166, 169, 171, 222, 233, 245, 246, 250, 251, 252, 256, 272, 273, 287, 292, 303, 314, 318, 320, 324, 336, 342, 343, 344, 345, 346, 349, 359, 360, 374, 408, 409, 415, 417, 424, 429, 430, 458, 464, 474 The antisense oligonucleotide according to claim 2, having 100% identity to a sequence selected from the group consisting of 490, 493, 512, 519, 529, 533, 534, 547, 566, 567, 578, 582, 601, 619, 620, 636, 637, 638, 645, 650, 651, 652, 653, 658, 659, 660, 665, 678, 679, 680, 682, 683, 684, 687, 694, 706, 716, 728, 733, 734, and 735.
  4. An antisense oligonucleotide conjugate comprising an oligonucleotide according to any one of claims 1 to 3 and a conjugate portion covalently bonded to the oligonucleotide.
  5. The antisense oligonucleotide conjugate according to claim 4, comprising a linker between the oligonucleotide and the conjugate portion.
  6. The antisense oligonucleotide conjugate according to claim 4 or 5, wherein the conjugate portion is an asialoclycoprotein receptor targeting portion.
  7. The antisense oligonucleotide conjugate according to claim 6, wherein the asialocrypoprotein receptor targeting portion is a trivalent N-acetylgalactosamine (GalNAc) portion.
  8. The antisense oligonucleotide conjugate according to any one of claims 5 to 7, wherein the linker is a physiologically unstable linker.
  9. The antisense oligonucleotide conjugate according to claim 8, wherein the physiologically unstable linker is a nuclease-sensitive linker.
  10. The antisense oligonucleotide conjugate according to claim 8 or 9, wherein the physiologically unstable linker comprises a cytidine-adenosine dinucleotide.
  11. The antisense oligonucleotide conjugate according to claim 4, wherein a linker is provided between the oligonucleotide and the conjugate portion, the conjugate portion comprises an asialocrycoprotein receptor targeting portion which is a trivalent N-acetylgalactosamine (GalNAc) moiety, the linker is a physiologically unstable linker, and the physiologically unstable linker further comprises a cytidine-adenosine dinucleotide.
  12. A pharmaceutical composition comprising an antisense oligonucleotide according to claims 1 to 3, or an antisense oligonucleotide conjugate according to any one of claims 4 to 11, and a pharmaceutically acceptable diluent, solvent, carrier, salt, and/or adjuvant.
  13. The pharmaceutical composition according to claim 12, wherein the pharmaceutically acceptable diluent is sterile phosphate-buffered saline.
  14. The pharmaceutical composition according to claim 12 or 13, wherein the pharmaceutically acceptable salt is a sodium salt.
  15. The pharmaceutical composition according to claim 12 or 13, wherein the pharmaceutically acceptable salt is a potassium salt.
  16. An in vivo or in vitro method for regulating PD-L1 expression in target cells expressing PD-L1, comprising administering an effective amount to the cells an antisense oligonucleotide according to any one of claims 1 to 3, an antisense oligonucleotide conjugate according to any one of claims 4 to 11, or a pharmaceutical composition according to any one of claims 12 to 15.
  17. An antisense oligonucleotide according to any one of claims 1 to 3, an antisense oligonucleotide conjugate according to any one of claims 4 to 11, or a pharmaceutical composition according to any one of claims 12 to 15, used for restoring an immune response to a virus.
  18. The antisense oligonucleotide, antisense oligonucleotide conjugate, or pharmaceutical composition according to claim 17, wherein the virus is hepatitis B virus (HBV).
  19. An antisense oligonucleotide according to any one of claims 1 to 3, an antisense oligonucleotide conjugate according to any one of claims 4 to 11, or a pharmaceutical composition according to any one of claims 12 to 15, for use in restoring the immune response to parasites.
  20. The antisense oligonucleotide, antisense oligonucleotide conjugate, or pharmaceutical composition according to any one of claims 17 to 19, wherein the recovery of the immune response is an increase in CD8+ T cells specific to one or more HBV antigens in the liver compared to a control.

Description

This invention relates to oligonucleotides (oligomers) that are complementary to programmed death ligand-1 (PD-L1) and reduce PD-L1 expression in the liver. The invention also relates to a method for alleviating T cell exhaustion caused by liver infection or cancer in the liver. Relevant infections include chronic HBV, HCV, HDV, and parasitic infections, such as malaria and toxoplasmosis (caused by protozoa of Plasmodium falciparum, particularly Plasmodium vivax, Plasmodium malariae, and Plasmodium falciparum). The costimulatory pathway consisting of the programmed death-1 (PD-1) receptor and its ligand, PD-L1 (or B7-H1 or CD274), is known to directly contribute to T cell exhaustion, resulting in a lack of viral control during chronic liver infections. The PD-1 pathway is also involved in autoimmunity when mice with disruption in this pathway develop autoimmune diseases. It has been shown that antibodies blocking the interaction between PD-1 and PD-L1 enhance the T cell response, particularly the response of CD8+ cytotoxic T cells (see Barber et al 2006 Nature Vol 439 p682 and Maier et al 2007 J. Immunol. Vol 178 p 2714). International Publication No. 2006/042237 describes a diagnostic method for cancer by evaluating the expression of PD-L1 (B7-H1) in tumors, and suggests delivering drugs that interfere with the PD-1/PD-L1 interaction to patients. These interfering agents can be antibodies, antibody fragments, siRNA, or antisense oligonucleotides. However, no specific examples of such interfering agents are provided, nor is there any mention of chronic liver infections. RNA interference-mediated inhibition of PD-L1 using double-stranded RNA (dsRNA, RNAi, or siRNA) molecules is also disclosed, for example, in International Publication No. 2005/007855, International Publication No. 2007/084865, and U.S. Patent No. 8,507,663. None of these describe targeted delivery to the liver. Dolina et al. (2013, Molecular Therapy-Nucleic Acids, 2 e72) describes the in vivo delivery of siRNA-targeting PD-L1 to Kupffer cells, thereby enhancing NK and CD8+ T cell clearance in MCMV-infected mice. This paper concludes that siRNA-targeting PD-L1 delivered to hepatocytes is not effective in enhancing CD8+ T cell effector function. The siRNA method differs considerably from the single-stranded antisense oligonucleotide method because their in vivo distribution and mode of action are completely different. As described by Xu et al. 2003 Biochem. Biophys. Res. Comm. Vol 306 pp 712-717, antisense oligonucleotides and siRNAs have different priorities for targeting sites in mRNA. International Publication No. 2016/138278 describes the inhibition of immune checkpoints, including PD-L1, using two or more single-stranded antisense oligonucleotides ligated to their 5' ends. This application does not refer to hepatitis B virus (HBV) or targeted delivery to the liver. This invention identifies novel oligonucleotides and oligonucleotide conjugates that highly efficiently reduce intracellular PD-L1 mRNA in both parenchymal cells (e.g., hepatocytes) and hepatocytes, as well as non-parenchymal cells such as Kupffer cells and hepatic sinusoidal endothelial cells (LSECs). By reducing or silencing PD-L1, the oligonucleotides and oligonucleotide conjugates reduce PD-1-mediated inhibition, thereby promoting immune stimulation of exhausted T cells. By mitigating T cell exhaustion in chronic hepatic pathogenic infection, a decrease in serum viral antigen levels and restoration of immune regulation during chronic hepatic pathogenic infection are achieved. Natural killer (NK) cells and natural killer T (NKT) cells are also considered to be activatable by the oligonucleotides and oligonucleotide conjugates of this invention. Oligonucleotide conjugates reliably achieve local reduction of PD-L1 in hepatocytes, thereby reducing the risk of autoimmune side effects such as pneumonia, nonviral hepatitis, and colitis associated with systemic PD-L1 depletion. This invention relates to oligonucleotides or conjugates that target nucleic acids capable of regulating PD-L1 expression, and to the treatment or prevention of diseases related to PD-L1 function. Oligonucleotides or oligonucleotide conjugates may be particularly used to treat diseases in which the immune response against infectious agents is exhausted. Therefore, in a first embodiment, the present invention provides an oligonucleotide comprising a continuous nucleotide sequence of 10 to 30 nucleotides in length, having at least 90% complementarity to the PD-L1 target nucleic acid. The oligonucleotide may preferably be an antisense oligonucleotide having a gapmer design. It is preferable that the oligonucleotide can inhibit PD-L1 expression by cleaving the target nucleic acid. Cleavage is preferably achieved by nuclease supplementation. In a further embodiment, the oligonucleotide is bound to at least one asialoglycoprotein receptor targeting binding moiety, for example, a binding moiety containing at least