Fusion Protein Containing BDNF

Patent metadata

Field	Value
Publication number	US-2021309754-A1
Application number	US-202117349414-A
Country	US
Kind code	A1
Filing date	Jun 16, 2021
Priority date	Jun 24, 2015
Publication date	Oct 7, 2021
Grant date	—

Abstract

Disclosed is a fusion protein containing a brain-derived neurotrophic factor (BDNF). The fusion protein is a fusion protein of BDNF and a specific range of human anti-transferrin receptor antibody, which makes BDNF administered into the blood able to pass through the blood-brain barrier.

First claim

1 - 73 . (canceled)

74 . A nucleotide sequence encoding a fusion protein of brain-derived neurotrophic factor (BDNF) and an anti-human transferrin receptor antibody (anti-hTfR),
wherein the fusion protein comprises a BDNF linked to an anti-hTfR antibody or an antigen-binding fragment thereof comprises a light chain (LC) having a light chain variable region (LCVR) comprising three complementarity-determining regions (LCDR1-3) and a heavy chain (HC) having a heavy chain variable region (HCVR) comprising three complementarity-determining regions (HCDR1-3), wherein the LCDR1 comprises the amino acid sequence of SEQ ID NO:16, the LCDR2 comprises the amino acid sequence of SEQ ID NO:18, the LCDR3 comprises the amino acid sequence of SEQ ID NO:20, the HCDR1 comprises the amino acid sequence of SEQ ID NO:88, the HCDR2 comprises the amino acid sequence of SEQ ID NO:90, and the HCDR3 comprises the amino acid sequence of SEQ ID NO:92, and wherein the BDNF is linked directly or via a linker to the HC of the anti-hTfR antibody or antigen-binding fragment thereof and the C-terminus of the BDNF is linked to the N-terminus of the HC.

75 . The nucleotide sequence according to claim 74 , wherein the linker is a peptide consisting of 1 to 50 amino acid residues.

76 . The nucleotide sequence according to claim 74 , wherein the linker is a peptide comprising the amino acid sequence selected from the group consisting of the amino acid sequence (Gly-Ser), the amino acid sequence (Gly-Gly-Ser), SEQ ID NO:3, SEQ ID NO:4, and SEQ ID NO:5.

77 . The nucleotide sequence according to claim 74 , wherein the BDNF is human BDNF.

78 . The nucleotide sequence according to claim 77 , wherein the human BDNF comprises the amino acid sequence having an identity not lower than 97% to the amino acid sequence set forth as SEQ ID NO:247, or
the human BDNF comprises the amino acid sequence having an identity not lower than 97% to the amino acid sequence set forth as SEQ ID NO:256.

79 . The nucleotide sequence according to claim 74 , having an affinity to both the extracellular region of human transferrin receptor and the extracellular region of monkey transferrin receptor.

80 . The nucleotide sequence according to claim 79 , wherein the dissociation constant of the anti-transferrin receptor antibody with the extracellular region of human transferrin receptor is not greater than 1×10-8 M, and the dissociation constant of the anti-transferrin receptor antibody with the extracellular region of monkey transferrin receptor is not greater than 5×10-8 M.

81 . A nucleotide sequence encoding a fusion protein of brain-derived neurotrophic factor (BDNF) and an anti-human transferrin receptor (anti-hTfR) scFv antibody, wherein the fusion protein comprises BDNF linked to the anti-hTfR scFv antibody with a linker, wherein the anti-hTfR scFv antibody comprises a light chain variable region (LCVR) comprising three complementarity-determining regions (LCDR1-3) and a heavy chain variable region (HCVR) comprising three complementarity-determining regions (HCDR1-3), wherein the LCDR1 comprises the amino acid sequence SEQ ID NO:16, the LCDR2 comprises the amino acid sequence of SEQ ID NO:18, the LCDR3 comprises the amino acid sequence of SEQ ID NO:20, the HCDR1 comprises the amino acid sequence SEQ ID NO:88, the HCDR2 comprises the amino acid sequence SEQ ID NO:90, and the HCDR3 comprises the amino acid sequence SEQ ID NO:92, wherein the BDNF is a human BDNF, and wherein the C-terminus of the BDNF is linked to the N-terminus of the HCVR via a first linker and the C-terminus of the HCVR is linked to the N-terminus of the LCVR via a second linker.

82 . The nucleotide sequence according to claim 81 , wherein the linker sequence consists of 2 to 50 amino acid residues.

83 . The nucleotide sequence according to claim 82 , wherein the linker sequence comprises the amino acid sequence selected from the group consisting of the amino acid sequence (Gly-Ser), the amino acid sequence (Gly-Gly-Ser), the amino acid sequence (Gly-Gly-Gly), SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and the amino acid sequence consisting of 2 to 10 thereof that are consecutively linked.

84 . The nucleotide sequence according to claim 81 , wherein the anti-hTfR scFv antibody consists of the amino acid sequence of SEQ ID NO:257.

85 . The nucleotide sequence according to claim 84 , wherein the anti-hTfR scFv antibody consists of the amino acid sequence of SEQ ID NO:257, the human BDNF is human pro-BDNF, and the fusion protein comprises the amino acid sequence set forth as SEQ ID NO:259.

86 . The nucleotide sequence according to claim 84 , wherein anti-hTfR scFv antibody consists of the amino acid sequence of SEQ ID NO:257, and the fusion protein comprises the amino acid sequence of SEQ ID NO:260.

87 . The nucleotide sequence according to claim 74 , wherein the antigen-binding fragment is a Fab, F(ab′)2, or F(ab′) of the anti-hTfR antibody and the BDNF is human BDNF; wherein the human BDNF is linked, directly or via a linker, to the heavy chain of the Fab, F(ab′)2, and F(ab′).

88 . The nucleotide sequence according to claim 87 , wherein the light chain of the antigen-binding fragment consists of the amino acid sequence of SEQ ID NO:196, and the heavy chain of the antigen-binding fragment consists of the amino acid sequence of SEQ ID NO:261.

89 . The nucleotide sequence according to claim 87 , wherein the light chain of the antigen-binding fragment consists of the amino acid sequence of SEQ ID NO:196, and the BDNF linked to the heavy chain of the antigen-binding fragment consists of the amino acid sequence of set forth as SEQ ID NO:263.

90 . The nucleotide sequence according to claim 87 , wherein the light chain of the antigen-binding fragment consists of the amino acid sequence of SEQ ID NO:196, and the BDNF linked to light chain of the antigen binding fragment consists of the amino acid sequence of SEQ ID NO:264.

91 . The nucleotide sequence according to claim 74 , wherein the LCVR comprises the amino acid sequence of SEQ ID NO:191; and the HCVR comprises the amino acid sequence of SEQ ID NO:205.

92 . The nucleotide sequence according to claim 81 , wherein the HCVR comprises the amino acid sequence of SEQ ID NO:205.

93 . The nucleotide sequence according to claim 74 , wherein the LCVR comprises the amino acid sequence having at least 95% sequence identity to SEQ ID NO:191 and having the LCDR 1-3 of SEQ ID NO:16, 18, and 20 respectively, and HCVR comprises the amino acid sequence having at least 95% sequence identity to SEQ ID NO:205 and having the HCDR 1-3 of SEQ ID NO:88, 90, and 92 respectively.

94 . The nucleotide sequence according to claim 93 , wherein LCVR comprises the amino acid sequence having at least 98% sequence identity to SEQ ID NO:191 and having the LCDR 1-3 of SEQ ID NO:16, 18, and 20 respectively, and HCVR comprises the amino acid sequence having at least 98% sequence identity to SEQ ID NO:205 and having the HCDR 1-3 of SEQ ID NO:88, 90, and 92 respectively.

95 . The nucleotide sequence according to claim 93 , wherein the LCVR comprises the amino acid sequence of SEQ ID NO:191 with substitution, deletion, or addition of 1 to 3 amino acids and has the LCDR 1-3 of SEQ ID NO:16, 18, and 20 respectively, or wherein the HCVR comprises the amino acid sequence of SEQ ID NO:205 with substitution, deletion, or addition of 1 to 3 amino acids and has the HCDR 1-3 of SEQ ID NO:88, 90, and 92 respectively.

96 . An expres

Assignees

• Japan Chem Res

Inventors

• Sonoda Hiroyuki
• Takahashi Kenichi

Classifications

• C07K2317/54

  F(ab')2 · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• A61K39/395

  Antibodies (agglutinins A61K38/36 {; as drug carriers A61K47/50}); Immunoglobulins; Immune serum, e.g. antilymphocytic serum · CPC title

• C07K16/2881 Primary

  against CD71 · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title