JP-7857306-B2 - Activin and tumor necrosis factor-alpha bifunctional antagonist and its 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 activin or an activin-related ligand, The isolated bifunctional antagonist molecule simultaneously neutralizes TNF signaling and activin signaling. The isolated bifunctional antagonist molecule is selected from the group consisting of: a bifunctional molecule having the heavy chain amino acid sequence described in SEQ ID NO: 39 and the light chain amino acid sequence described in SEQ ID NO: 17; a bifunctional molecule having the heavy chain amino acid sequence described in SEQ ID NO: 41 and the light chain amino acid sequence described in SEQ ID NO: 17; a bifunctional molecule having the amino acid sequence described in SEQ ID NO: 60; and a bifunctional molecule having the amino acid sequence described in SEQ ID NO: 49. 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 Applications This application claims the interests of U.S. Provisional Application No. 63/113,918, filed on 15 November 2020, and U.S. Provisional Application No. 63/104,765, filed on 23 October 2020, each of which is incorporated herein by reference in whole. 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. As a subset of the TGF-β superfamily, activins, including activin A, activin B, and activin AB, and activin-related proteins, including myostatin (GDF-8) and GDF-11, mediate Smad2/3 signaling through the binding and activation of their high-affinity receptors, ActRIIA and ActRIIB, on the cell surface. Activins and related proteins play essential roles in regulating a wide range of biological activities, including mesoderm induction, cell differentiation, myogenesis, bone remodeling, hematopoiesis, fibrosis, and reproductive physiological functions. The secreted glycoprotein, follistatin (FST), binds to activins and activin-related ligands, negatively regulating their signaling activity. Overexpression of activin and related ligands, along with increased Smad2/3 signaling, has been linked to the pathogenesis and progression of many distressing conditions, such as cancer, anemia, bone metastases, bone fragility, fractures, muscle wasting disorders, cachexia, pulmonary hypertension, fibrosis, pain, insulin resistance, chronic kidney disease, liver disease, myocardial infarction, and heart failure. Increasing evidence indicates that many complex disorders involve the parallel activation of the TNF-α-mediated NF-κB signaling pathway and the activin-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 activin-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 activin 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-binding polypeptide") that specifically binds to a TNF-α ligand and a second antigen-binding molecule ("activin-binding polypeptide") that specifically binds to activin or an activin-related ligand. In various embodiments, the "TNF-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, while the "a