JP-7857307-B2 - Bifunctional antagonists of tumor necrosis factor-alpha and transforming growth factor-beta, and their use

Inventors

• ハン，ハック
• シュウ，シャオラン

Assignees

• ハン，ハック
• シュウ，シャオラン

Dates

Publication Date

20260512

Application Date

20211022

Priority Date

20201023

Claims (6)

1. An isolated bifunctional antagonist molecule comprising a first antigen-binding molecule that specifically binds to a TNF ligand and a second antigen-binding molecule that specifically binds to a TGF-β ligand, The isolated bifunctional antagonist molecule simultaneously neutralizes TNF signaling and TGF-β signaling. The isolated bifunctional antagonist molecule is a bifunctional antagonist molecule having the heavy chain amino acid sequence described in SEQ ID NO: 31 and the light chain amino acid sequence described in SEQ ID NO: 11. An isolated, bifunctional antagonist molecule characterized by the following features .
2. A pharmaceutical composition comprising a therapeutically effective amount of the bifunctional antagonist molecule according to claim 1, mixed with a pharmaceutically acceptable carrier.
3. An isolated nucleic acid molecule comprising a polynucleotide encoding the bifunctional antagonist molecule described in claim 1 .
4. A recombinant vector comprising the nucleic acid molecule described in claim 3 .
5. A host cell comprising the recombinant vector described in claim 4 .
6. A method for producing a bifunctional antagonist molecule according to claim 1 , comprising the steps of: a) transforming host cells with a vector comprising a polynucleotide encoding the bifunctional antagonist molecule; b) culturing the host cells under conditions suitable for the expression of the bifunctional antagonist molecule; and c) recovering the bifunctional antagonist molecule from the culture.

Description

Related Patent Application This application claims the benefit of U.S. Provisional Application No. 63/104,850, filed on 23 October 2020, which is incorporated in its entirety herein by reference. Background Technology Tumor necrosis factor-alpha (TNF-α) mediates NF-κB signaling and plays a crucial role in various physiological and pathological processes, including cell proliferation, differentiation, apoptosis, and modulation of immune responses and induction of inflammation. TNF acts through two receptors, TNFR1 (TNF receptor-1) and TNFR2 (TNF receptor-2). TNF-α plays a critical role in inflammatory responses, programmed cell death, and tissue necrosis. Increased TNF-α signaling has been linked to numerous inflammatory diseases, including rheumatoid arthritis, ankylosing spondylitis, Crohn's disease, and psoriasis, and anti-TNF therapies, such as adalimumab, infliximab, and etanercept, have been shown to be highly effective in treating such inflammatory diseases. Elevated TNF-α levels and increased TNF-α signaling have also been linked to the pathogenesis and progression of many other disease conditions, including anemia, leukemia, multiple myeloma, fibrosis, hypertension, muscle wasting, osteopenia, neurodegeneration, sepsis, pain, chronic kidney disease, liver disease, and heart failure. Transforming growth factor-beta (TGF-β), including TGF-β1, TGF-β2, and TGF-β3, mediates Smad2/3 signaling through the binding and activation of high-affinity receptors TGFβRII and TGFβRIIB on the cell surface. TGF-β plays a crucial role in regulating a wide range of biological activities, including immune function, cell proliferation and differentiation, epithelial-mesenchymal transition, fibrosis, hematopoiesis, myogenesis, and bone remodeling. Elevated TGF-β levels and resulting increased Smad2/3 signaling have been linked to the pathogenesis and progression of numerous disease conditions, including cancer, anemia, bone metastases, osteopenia, fibrosis, pain, muscle loss, insulin resistance, chronic kidney disease, liver disease, and cardiovascular disease. Increasing evidence indicates that many complex disorders involve the parallel activation of the TNF-α-mediated NF-κB signaling pathway and the TGF-β-mediated Smad2/3 signaling pathway, whose activity promotes pathogenesis and progression. Examples of such complex disorders include certain hematological disorders, e.g., refractory anemia and myelodysplastic syndromes; cardiovascular diseases, e.g., pulmonary hypertension and congestive heart failure; bone disorders, e.g., bone metastases and fractures; organ failure, e.g., renal failure, hepatic failure, or bone marrow failure; fibrous diseases, e.g., non-alcoholic steatohepatitis, cirrhosis, and pulmonary fibrosis; and pain, e.g., nociceptive or neuropathic pain. Current treatment options for these complex disorders are limited. Due to the involvement of one or more disease signaling mechanisms in these disorders, currently available therapies designed to target a single disease mechanism typically exhibit poor efficacy and low response rates. Since both the TNF-α-NF-κB signaling pathway and the TGF-β-Smad2/3 signaling pathway are essentially involved in disease pathogenesis and progression, developing novel bifunctional antagonists capable of inhibiting both disease signaling pathways in parallel is clearly important. Disclosure of the Invention In one embodiment, the present invention provides a novel polypeptide-based bifunctional antagonist molecule specifically designed to simultaneously neutralize TNF-α signaling and TGF-β signaling in a potent manner. In various embodiments, the bifunctional antagonist molecule is designed as depicted in Figure 1. In various embodiments, the bifunctional antagonist molecule is designed as depicted in Figure 2. In various embodiments, the bifunctional antagonist molecule is designed as depicted in Figure 3. In various embodiments, the bifunctional antagonist molecule is a bifunctional molecule comprising a first antigen-binding molecule ("TNF-α-binding polypeptide") that specifically binds to a TNF-α ligand and a second antigen-binding molecule ("TGF-β-binding polypeptide") that specifically binds to a TGF-β ligand. In various embodiments, the "TNF-α-binding polypeptide" is selected from the group consisting of anti-TNF antibodies, fragments of anti-TNF antibodies, wild-type TNFR1 and TNFR2 extracellular domains (ECDs), modified TNFR1 and TNFR2 extracellular domains, and phage display-derived polypeptides that target TNF-α ligands; and the "TGF-β-binding polypeptide" is selected from the group consisting of anti-TGF-β antibodies, fragments of anti-TGF-β antibodies, wild-type TGF-β2 receptor (including TGFβRIIA and TGFβRIIB) extracellular domains (ECDs), modified TGFβRIIA and TGFβRIIB extracellular domains, and phage display-derived antagonist polypeptides that target TGF-β ligands. In various embodiments, the bifunctional molecule comprises an isolated antibody or its antigen