JPWO2020180819A5

JPWO2020180819A5 -

Info

Publication number: JPWO2020180819A5
Application number: JP2021552257A
Authority: JP
Publication date: 2023-03-09

Claims

An isolated antibody that specifically binds to human tau, comprising:
a mature heavy chain variable region comprising CDR-H1 comprising SEQ ID NO:8, CDR-H2 comprising SEQ ID NO:9 or SEQ ID NO:149, and CDR-H3 comprising SEQ ID NO:10 and which is at least 90% identical to SEQ ID NO:18 ,
CDR-L1 comprising SEQ ID NO: 12, CDR- comprising SEQ ID NO: 150, 151, 153, 156, 158, 159, 160, 163, 165, 166, 167, 168, 169, 170, 171, 172, 173 or 174 a mature light chain variable region that is at least 90% identical to SEQ ID NO: 122, comprising L2, and CDR-L3 comprising SEQ ID NO: 14;
Antibodies, including

at least one of positions H12, H13, H17, H24, H40, H43, H48, H66, H67, H76, H80, H81, and H91 are V, K, T, A, R, Q, I, respectively; R, A, D, L, Q, and F , and at least one of positions L2, L12, L15, L37, L39, L45, L60 and L100 are V, P, L, Q, respectively. , R, R, D and Q.

2. The antibody of claim 1, wherein CDR-L2 comprises SEQ ID NO: 150, 151, 163, 167, 168, or 169.

4. The antibody of claim 3, wherein said heavy chain variable region comprises SEQ ID NO: 18 and said light chain variable region comprises SEQ ID NO: 110, 121, 122 or 123.

5. The antibody of claim 4, wherein said light chain variable region comprises SEQ ID NO:122.

4. The antibody of claim 3, wherein said heavy chain variable region comprises SEQ ID NO:146 and said light chain variable region comprises SEQ ID NO:94.

4. The antibody of claim 3, wherein said heavy chain variable region comprises SEQ ID NO:146 and said light chain variable region comprises SEQ ID NO:122.

The antibody of any one of claims 1-7 , which is a chimeric antibody, a veneered antibody, or a humanized antibody.

The antibody of any one of claims 1-8 , which is a whole antibody.

The antibody of any one of claims 1-8 , which is an antigen- binding fragment.

11. The antibody of claim 10 , wherein said antigen binding fragment is a single chain antibody, Fab, or _Fab'2 fragment.

The antibody of any one of claims 1-11 , whose isotype is human IgG1 , human IgG2, or human IgG4 .

13. The antibody of any one of claims 1-9 and 12 , wherein said mature light chain variable region is fused to a light chain constant region and said mature heavy chain variable region is fused to a heavy chain constant region. body.

14. The antibody of claim 13 , wherein said heavy chain constant region has the amino acid sequence of SEQ ID NO: 176 with or without a C-terminal lysine .

14. The antibody of claim 13 , wherein the mature heavy chain variable region fused to the heavy chain constant region has the amino acid sequence of SEQ ID NO: 178 with or without a C-terminal lysine .

14. The antibody of claim 13 , further comprising a signal peptide fused to said mature heavy chain variable region and/or said mature light chain variable region.

16. The antibody of claim 15 , wherein the heavy chain has the amino acid sequence of SEQ ID NO: 180 with or without a C-terminal lysine .

14. The antibody of claim 13 , wherein said light chain constant region has the amino acid sequence of SEQ ID NO:177.

14. The antibody of claim 13 , wherein the mature light chain variable region fused to the light chain constant region has the amino acid sequence of SEQ ID NO:179.

20. The antibody of Claim 19 , wherein the light chain has the amino acid sequence of SEQ ID NO:181.

16. The antibody of claim 15 , wherein the heavy chain has the amino acid sequence of SEQ ID NO:178 with or without a C-terminal lysine and the light chain has the amino acid sequence of SEQ ID NO:179.

18. The antibody of claim 17 , wherein the heavy chain has the amino acid sequence of SEQ ID NO:180 with or without a C-terminal lysine and the light chain has the amino acid sequence of SEQ ID NO:181.

23. The antibody of any one of claims 1-22 , which has at least one mutation in the constant region.

24. The antibody of claim 23 , wherein said mutation reduces complement fixation or activation by said constant region , or reduces binding to an Fc receptor , compared to a native human heavy chain constant region. .

having a mutation at one or more of positions 241, 264, 265, 270, 296, 297, 318, 320, 322, 329 and 331 by EU numbering , or having an alanine at positions 318, 320 and 322; 25. The antibody of claim 24 .

26. The antibody of any one of claims 1-25 , which is conjugated to a therapeutic, cytotoxic, cytostatic, neurotrophic, or neuroprotective agent.

A pharmaceutical composition comprising the antibody of any one of claims 1-26 and a pharmaceutically acceptable carrier .

A nucleic acid encoding the heavy and/or light chain of the antibody of any one of claims 1-26 .

29. The nucleic acid of claim 28, wherein said heavy chain is encoded by a sequence comprising SEQ ID NO:182 and said light chain is encoded by a sequence comprising SEQ ID NO: 183 .

A method of producing an antibody comprising:
(a) culturing cells transformed with the nucleic acid of claim 29 such that the cells secrete said antibody; and (b) purifying said antibody from the cell culture medium.
A method, including

A method of producing an antibody -producing cell line comprising:
(a) introducing into a cell a vector encoding the nucleic acid of claim 29 and a selectable marker ;
(b) growing said cells under conditions to select for cells with increased copy number of said vector;
(c ) isolating single cells from the selected cells; and (d) banking cloned cells from the single cells selected based on antibody yield.
A method, including

A method of producing an antibody comprising:
(a) culturing cells transformed with nucleic acids encoding the heavy and light chains of the antibody of claim 1, such that the cells secrete said antibody; and (b) from cell culture medium said purifying the antibody;
A method, including

A method of producing an antibody -producing cell line comprising:
(a) introducing into a cell a vector encoding the heavy and light chains of the antibody of claim 1 and a selectable marker;
(b) growing said cells under conditions to select for cells with increased copy number of said vector;
(c ) isolating single cells from the selected cells; and (d) banking cloned cells from the single cells selected based on antibody yield.
A method, including

27. A method of inhibiting or reducing tau aggregation in a subject with tau-mediated amyloidosis or at risk of developing tau-mediated amyloidosis , comprising: A method comprising administering an effective dosing regimen to said subject, thereby inhibiting or reducing tau aggregation in said subject.

35. The method of claim 34 , wherein said antibody is a humanized version of an antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:7 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:11. .

27. A method of treating a tau-related disease or effecting prophylaxis of a tau-related disease in a subject, comprising administering an effective dosing regimen of an antibody as defined in any one of claims 1-26 and thereby preventing said disease. or practicing prophylaxis of said disease .

A method of reducing tau abnormal propagation , inducing tau phagocytosis, inhibiting tau deposition or aggregation, or inhibiting tau tangle formation in a subject , the method comprising : administering an effective dosing regimen of an antibody as defined in any and thereby reducing tau propagation , inducing tau phagocytosis, inhibiting tau deposition or aggregation, or reducing tau in said subject; A method comprising inhibiting the formation of tangles .

35. The method of claim 34 , wherein said antibody is administered into said subject's body by intravenous injection.

A method of measuring efficacy of treatment in a subject treated for a disease associated with tau aggregation or deposition, comprising:
(a) pretreatment by administering an antibody as defined in any one of claims 1 to 26 to a subject and detecting a first amount of antibody bound to tau in said subject; measuring a first level of tau protein deposits in said subject;
(b) administering said treatment to said subject;
(c) measuring a second level of tau protein deposits in said subject after treatment by administering said antibody to said subject and detecting antibody bound to tau in said subject;
including
a reduction in the level of tau protein deposits indicates a positive response to treatment;
Method.

A method of measuring efficacy of treatment in a subject treated for a disease associated with tau aggregation or deposition, comprising:
(a) pretreatment by administering an antibody as defined in any one of claims 1 to 26 to a subject and detecting a first amount of antibody bound to tau in said subject; measuring a first level of tau protein deposits in said subject;
(b) administering said treatment to said subject;
(c) a second level of tau protein deposits in said subject after treatment by administering said antibody to said subject and detecting a second amount of antibody bound to tau in said subject; the step of measuring
including
A method wherein no change in the level of tau protein deposits or a slight increase in tau protein deposits indicates a positive response to treatment.

A method of producing an antibody that specifically binds human tau at an epitope within a motif of the formula KXXSXXNX(K/H)H (SEQ ID NO: 191) or KIGSLDNITH (SEQ ID NO: 194), comprising immunizing an animal with human tau or a fragment thereof. and screening for one of the produced antibodies that specifically binds within said motif.

42. The method of claim 41 , wherein said animal is immunized with 383 amino acid human tau (4R0N).

43. The method of claim 42 , wherein said human tau comprises a P301S mutation.

wherein said screen comprises said antibody and any other antibody represented by KIGSTENLKH (SEQ ID NO: 188), KCGSKDNIKH (SEQ ID NO: 192), KCGSLGNIHH (SEQ ID NO: 193), or KXXSXXNX(K/H)H (SEQ ID NO: 191), respectively. 42. The method of claim 41 , wherein specific binding between one or more peptides of up to 15 amino acids comprising a consensus motif is determined.

42. The method of claim 41 , wherein said animal is immunized with a tau fragment of up to 15 amino acids comprising KXXSXXNX(K/H)H (SEQ ID NO: 191) linked to a carrier.

46. The method of claim 45 , wherein the peptide is KIGSTENLKH (SEQ ID NO: 188) or KCGSKDNIKH (SEQ ID NO: 192) or KCGSLGNIHH (SEQ ID NO: 193).