Trail compositions with reduced immunogenicity
Inventors
Sokoloff, Alex • Roschke, Viktor • Lee, Kang Choon • Lee, Seulki • Oh, Yumin
Assignees
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Publication Number
US-11767353-B2
Publication Date
2023-09-26
Expiration Date
Abstract
Fusion polypeptides with modified multimerization domains that provide high expression, solubility, stability, and low immunogenicity to the fusion polypeptides have been developed. TRAIL compositions with the modified multimerization domains show improved physico-chemical and biological properties relative to TRAIL compositions with unmodified multimerization domains. The TRAIL compositions also have lower immunogenicity in the mammalian host when compared to that of TRAIL compositions with unmodified multimerization domains. The TRAIL compositions induce apoptosis of cancer cells and cancer-associated fibroblasts in vivo, reducing tumor size.
Core Innovation
The invention relates to recombinant fusion polypeptides comprising a modified isoleucine zipper (miLZ) multimerization domain fused to a tumor necrosis factor-related apoptosis ligand (TRAIL) domain. The fusion polypeptides use miLZ sequences corresponding to SEQ ID NOs: 4-8 and TRAIL sequences corresponding to SEQ ID NOs: 9-26, including soluble extracellular fragments such as 114-281, and the multimerization and TRAIL domain arrangement forms trimeric fusion polypeptides/multimers.
A central aspect is modifying iLZ residues targeted within the miLZ sequences to preserve trimerization while reducing immunogenicity. The modifications are described in terms of selecting amino acids at positions 14, 17, and 26 of SEQ ID NOs: 4-8, and avoiding immunogenic epitopes of isoleucine at position 17 together with lysine at position 26, while maintaining the ability to trimerize.
The disclosure additionally states that the modified miLZ-TRAIL fusions are formulated or conjugated to improve developability, including improved solubility/stability and extended in vivo half-life. PEGylated conjugates are described with polyethylene glycol (PEG) molecular weight ranges of about 5,000 to about 100,000 Da, including mono- and di-PEGylated forms, and as improving solubility, stability to freeze-thaw, longer non-human primate half-life, and reduced immunogenicity. Examples include TLY012 as a PEGylated miLZ-TRAIL.
The problem addressed is the need for TRAIL-based therapeutic fusion polypeptides with reduced immunogenicity and improved properties such as solubility/stability and extended in vivo half-life, while maintaining or improving in vitro potency. The disclosure emphasizes apoptosis induction using the TRAIL domain, including in cancer cells and in cancer-associated fibroblasts/activated stellate cells, and includes therapeutic indication concepts for proliferative (cancer) and fibrotic/autoimmune diseases. The document further describes polynucleotides/expressing constructs, pharmaceutical compositions, and combination therapy concepts.
Claims Coverage
The partial claim set includes four independent claims across the items: two covering a recombinant fusion polypeptide with a first amino acid sequence selected from or having at least 90% identity to SEQ ID NOs: 4-8 and a second amino acid sequence comprising a TRAIL domain, one of which further imposes an immunogenicity-avoidance constraint; and two additional claims describing PD-1/PD-L1 antagonist modalities and related therapeutic combinations. Across these claims, the core inventive coverage includes the modified isoleucine zipper sequence family (SEQ ID NOs: 4-8) fused to a TRAIL domain, with one independent claim additionally requiring selection at positions 14, 17, and 26 to avoid a defined immunogenic epitope pattern.
Modified isoleucine zipper fused to a trail domain
A recombinant fusion polypeptide comprising a first amino acid sequence having at least 90% identity to any one of SEQ ID NOs: 4-8, and a second amino acid sequence comprising a TRAIL domain.
Immunogenicity-avoidance by amino-acid selection at positions 14, 17, and 26
At least one of the amino acids at positions 14, 17, and 26 of SEQ ID NOs: 4-8 is selected from Valine (V), Glutamic acid (E), and/or Glutamine (Q), and immunogenic epitopes of Isoleucine at position 17 and Lysine at position 26 are not present together in the first amino acid sequence.
Recombinant fusion polypeptide defined by miLZ and TRAIL domain selection
A recombinant fusion polypeptide comprising a first amino acid sequence selected from SEQ ID NOs: 4-8 and a second amino acid sequence comprising a TRAIL domain.
PD-1/pd-l1 antagonist modalities
Alternative PD-1/PD-L1 antagonist modalities in which immune-checkpoint inhibition is provided through recombinant fusion polypeptides and PD-1 ligand-based formats.
Immune-checkpoint inhibitor combinations
Additional immune-checkpoint antagonists and immune-checkpoint inhibitor combinations used with PD-1/PD-L1 antagonists in the same overall therapeutic framework.
Taken together, the independent claims cover miLZ (SEQ ID NOs: 4-8) fused to a TRAIL domain, with one independent claim further restricting the first sequence to meet an immunogenicity-avoidance constraint based on amino-acid selection at positions 14, 17, and 26 and the absence of immunogenic epitopes characterized by isoleucine at position 17 together with lysine at position 26. The other claimed matter describes PD-1/PD-L1 antagonist modalities and immune-checkpoint inhibitor combinations in the same therapeutic framework.
Stated Advantages
Reduced immunogenicity.
Improved solubility/stability.
Extended in vivo half-life.
Increased in vitro potency.
Stability to freeze-thaw.
Longer non-human primate half-life.
Documented Applications
Therapeutic use for proliferative (cancer) disease by administration of an effective amount of the recombinant fusion polypeptide of claim 1 or an effective amount of the conjugate.
Therapeutic use for autoimmune disease by administration of an effective amount of the recombinant fusion polypeptide of claim 1 or an effective amount of the conjugate.
Therapeutic use for fibrotic disease by administration of an effective amount of the recombinant fusion polypeptide of claim 1 or an effective amount of the conjugate.
Apoptosis induction in cancer cells and cancer-associated fibroblasts/activated stellate cells, as described for TRAIL-domain activity.
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