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CN-122003443-A - Fusion proteins

CN122003443ACN 122003443 ACN122003443 ACN 122003443ACN-122003443-A

Abstract

The present invention relates to a p75NTR neurotrophic factor binding protein (NBP) -Fc fusion protein comprising a p75NTR (NBP) portion and an immunoglobulin portion. In certain embodiments, the p75NTR (NBP) -Fc fusion protein is used to treat pain and/or symptoms of pain.

Inventors

  • S. Westbrook
  • J. Rutter

Assignees

  • 利维塞普特有限公司

Dates

Publication Date
20260508
Application Date
20240521
Priority Date
20230526

Claims (9)

  1. 1. A p75NTR neurotrophic factor binding protein (NBP) -Fc fusion protein comprising: (a) A p75NTR (NBP) moiety selected from one of Seq ID No.1-19, and (B) An immunoglobulin Fc portion selected from one of Seq ID No. 20-24; wherein the p75NTR (NBP) and Fc moiety are linked by a linker comprising a peptide of formula G x , wherein x is 1,2,3, 4, 5 or 6.
  2. 2. The p75NTR (NBP) -Fc fusion protein according to claim 1, wherein the linker is GGG.
  3. 3. The p75NTR (NBP) -Fc fusion protein according to claim 1 or 2, wherein the Fc region is an Fc suitable for human use.
  4. 4. Use of a p75NTR (NBP) -Fc fusion protein as described in any one of claims 1 to 3 for pain treatment.
  5. 5. A nucleic acid molecule encoding a p75NTR (NBP) -Fc fusion protein as described in any one of claims 1 to 3.
  6. 6. A replicable expression vector for transfecting a cell, said vector comprising the nucleic acid molecule as described in claim 5.
  7. 7. Use of a nucleic acid molecule as described in claim 5 or a vector as described in claim 6 for pain treatment.
  8. 8. Use of a p75NTR (NBP) -Fc fusion protein as described in any one of claims 1 to 3, a nucleic acid molecule as described in claim 5 or a vector as described in claim 6 for the treatment of osteoarthritis.
  9. 9. A pharmaceutical composition comprising a p75NTR (NBP) -Fc fusion protein as described in any one of claims 1 to 3, a nucleic acid molecule as described in claim 5 or a vector as described in claim 6, and a pharmaceutically acceptable carrier and/or excipient.

Description

Fusion proteins Background Neurotrophic factors, neurotrophic Growth Factor (NGF), brain Derived Neurotrophic Factor (BDNF), neurotrophic factor 3 (NT-3) and neurotrophic factor 4/5 (NT-4/5) act through four receptors, the low affinity p75 neurotrophic receptor (p 75 NTR) and the high affinity tyrosine kinase receptor, trkA, trkB and TrkC. The low affinity receptor p75NTR can bind to and be activated by all four neurotrophic factors, and its function is reported to be independent of other receptors. However, activation of the Trk receptor is more selective, i.e. NGF is a selective ligand for TrkA, BDNF is a ligand for TrkB, and NT-3, 4/5 are ligands for TrkC. In addition, p75NTR and Trk proteins have been reported to form complexes when co-expressed, thereby altering signaling of both receptors (Huang and Reichardt,2003,Annu Rev Biochem. 72:609-42). Indeed, studies have shown that p75NTR is able to promote the selectivity of each neurotrophic factor for the respective Trk receptor. P75NTR is a member of the tumor necrosis factor receptor superfamily (TNFR-SF) and is also the first well characterized member of this superfamily. The superfamily (which in humans is encoded by about 30 genes) is defined by a ligand binding domain consisting of one or more (typically 4) repeated 40 amino acid Cysteine Rich Domains (CRDs) originally found in p75NTR (Johnson et al, 1986 Cell 47:545-554; radeke et al, 1987 Nature 325:593-597). In contrast, the intracellular domains of all TNFR-SF family members do not share any sequence motifs. Therefore, the signaling mechanisms of TNFR-SF proteins vary widely. An unusual feature of the p75NTR structure is the presence of disulfide-linked p75NTR dimers formed by cysteine residues within the transmembrane domain. This disulfide bond is required for efficient neurotrophin-dependent signaling by p75NTR and plays an important role in the formation of intracellular and extracellular domains (Vilar et al, 2009 Neuron 62:72-83). Neurotrophins exist physiologically as non-covalently bound dimers (Bothwell and Shooter, 1977J Biol Chem.252 (23): 8532-6.) and have a half-life profile of about 5 minutes (Tria et al, 1994 Exp Neurol.127 (2): 178-83). Neurotrophin-dependent p75NTR activation involves binding of a neurotrophin dimer to CRD2-4 of the two extracellular domains of the p75NTR dimer (He and Garcia, 2004 Science 304:870-875). Recent studies support a model in which neurotrophin binding results in the two extracellular domains of the p75NTR dimer being close to each other, forcing the intracellular domains to open in a "snail tongue" like manner centered on disulfide bonds, thereby binding the intracellular domains to the signaling adapter proteins NRIF and TRAF6 (Vilar et al, 2009J Cell Sci 122:3351-3357, vilar et al, 2009 Neuron 62:72-83). Disulfide bonds within the transmembrane domain (e.g., those found in p75 NTR) have not previously been described in other TNFR-SF family members or in any other membrane proteins. The p75NTR is proteolytically cleaved by alpha-secretase and gamma-secretase activities and Matrix Metalloproteinases (MMP) in turn, releasing its intracellular domain (ICD) into the cytoplasm in a manner analogous to the cleavage-dependent signaling pathway of Notch and beta-amyloid precursors (Jung et al, 2003J Biol Chem 278:42161-42169; kanning et al, 2003J Neuro-sci 23:5425-5436). The cytoplasmic release of p75NTR ICD by this pathway promotes signaling of the relevant NRIF (KENCHAPPA et al, 2006 Neuron 50:219-232). The role of the p75NTR extracellular domain after proteolytic cleavage of the alpha-secretase and gamma-secretase activities and MMPs is not completely understood. NGF and other neurotrophic factors (BDNF, NT-3 and NT-4/5) have been shown to play important roles in pathology, such as pain caused by osteoarthritis, pancreatitis, rheumatoid arthritis, psoriasis, pruritus and multiple sclerosis (Watanabe et al, 2008J Neurosci Res.86 (16): 3566-74; raychaudhuri et al, 2011 Arthritis Rheum.63 (11): 3243-52; barthel et al, 2009 Arthritis Res Ther.11 (3): R82; truzzi et al, 2011 Cell Death Differ.18:948-58; mcDonald et al, 2011. 2011 Curr Med Chem:234-44; yamaoka et al, 2007J Dermatol Sci.46 (1): 41-51). Selective antibodies against any neurotrophic factor (NGF or BDNF, NT-3 and NT-4/5) have been shown to significantly reduce pain. Furthermore, antibodies against the neurotrophic factor receptor p75NTR Trk A, trk B or Trk C have also been shown to be effective in pain models (Orita S et al, 2010J Orthop Res, 28:1614-20; 2010 Pain:473-80; iwakura et al, 2010J Hand Surg Am.35:267-73; cirilio et al, 2010 Cell Mol Neurobiol.30:51-62; pezet et al, 2010 Pain.90:113-25; hayashi et al, 2011J Pain, 12:1059-68; chu et al, 2011 Pain, 152:1832-7; ueda et al, 2010J Pharmacol Sci, 112:438-43; ghilardi et al, 2010 Bone, 48:389-98; fukui et al, 2010J Orthop Res, 2010, 28:279-83). Fukui et al, (2010) demonstrated significant efficacy of anti-p 75NTR antibody treatment on pain-rel