Anti-Myostatin Antibodies, Polypeptides Containing Variant Fc Regions, and Methods of Use

Patent metadata

Field	Value
Publication number	US-2016200807-A1
Application number	US-201514974488-A
Country	US
Kind code	A1
Filing date	Dec 18, 2015
Priority date	Dec 19, 2014
Publication date	Jul 14, 2016
Grant date	—

Abstract

The disclosure provides anti-myostatin antibodies and methods of making and using the same. Nucleic acids encoding the anti-myostatin antibodies and host cells comprising the nucleic acids are also provided. The anti-myostatin antibodies have uses that include treating a muscle wasting disease, reducing body fat accumulation, and increasing mass and strength of muscle tissue. The disclosure also provides polypeptides containing a variant Fc region and methods of making and using the same. Nucleic acids encoding the polypeptides and host cells comprising the nucleic acids are also provided. The polypeptides have uses that include suppressing the activation of immune cells; treating an immunological inflammatory disease, autoimmune disease, or viral infection; and increasing muscle mass and strength or reducing body fat accumulation.

First claim

What is claimed is:

1 . An isolated antibody that binds to latent myostatin, wherein the antibody inhibits activation of myostatin.

2 . The antibody of claim 1 , wherein the antibody:
(a) blocks the release of mature myostatin from latent myostatin; (b) blocks the proteolytic release of mature myostatin; (c) blocks the spontaneous release of mature myostatin; (d) does not bind to mature myostatin; or (e) binds to an epitope within a fragment consisting of amino acids 21-100 of myostatin propeptide (SEQ ID NO: 78).

3 . The antibody of claim 1 , wherein the antibody competes for binding latent myostatin with, or binds the same epitope as, an antibody comprising a VH and VL pair described in Table 2a, 11a, or 13.

4 . The antibody of claim 1 which binds to latent myostatin with higher affinity at neutral pH than at acidic pH, or which binds to latent myostatin with higher affinity at pH7.4 than at pH5.8.

5 . The antibody of claim 1 which is (a) a monoclonal antibody; (b) a human, humanized, or chimeric antibody; (c) a full length IgG antibody; or (d) an antibody fragment that binds to myostatin.

6 . The antibody of claim 1 , wherein the antibody comprises:
(a) (i) a HVR-H3 comprising the amino acid sequence GVPAX 1 SX 2 GGDX 3 , wherein X 1 is Y or H, X 2 is T or H, X 3 is L or K (SEQ ID NO: 128), (ii) a HVR-L3 comprising the amino acid sequence AGGYGGGX 1 YA, wherein X 1 is L or R (SEQ ID NO: 131), and (iii) a HVR-H2 comprising the amino acid sequence IISX 1 AGX 2 X 3 YX 4 X 5 X 6 WAKX 7 , wherein X 1 is Y or H, X 2 is S or K, X 3 is T, M or K, X 4 is Y or K, X 5 is A, M or E, X 6 is S or E, X 7 is G or K (SEQ ID NO: 127); (b) (i) a HVR-H1 comprising the amino acid sequence X 1 X 2 DIS, wherein X 1 is S or H, X 2 is Y, T, D or E (SEQ ID NO: 126), (ii) a HVR-H2 comprising the amino acid sequence IISX 1 AGX 2 X 3 YX 4 X 5 X 6 WAKX T , wherein X 1 is Y or H, X 2 is S or K, X 3 is T, M or K, X 4 is Y or K, X 5 is A, M or E, X 6 is S or E, X 7 is G or K (SEQ ID NO: 127), and (iii) a HVR-H3 comprising the amino acid sequence GVPAX 1 SX 2 GGDX 3 , wherein X 1 is Y or H, X 2 is T or H, X 3 is L or K (SEQ ID NO: 128); (c) (i) a HVR-H1 comprising the amino acid sequence X 1 X 2 DIS, wherein X 1 is S or H, X 2 is Y, T, D or E (SEQ ID NO: 126), (ii) a HVR-H2 comprising the amino acid sequence IISX 1 AGX 2 X 3 YX 4 X 5 X 6 WAKX 7 , wherein X 1 is Y or H, X 2 is S or K, X 3 is T, M or K, X 4 is Y or K, X 5 is A, M or E, X 6 is S or E, X 7 is G or K (SEQ ID NO: 127), (iii) a HVR-H3 comprising the amino acid sequence GVPAX 1 SX 2 GGDX 3 , wherein X 1 is Y or H, X 2 is T or H, X 3 is L or K (SEQ ID NO: 128), (iv) a HVR-L1 comprising the amino acid sequence X 1 X 2 SQX 3 VX 4 X 5 X 6 NWLS, wherein X 1 is Q or T, X 2 is S or T, X 3 is S or E, X 4 is Y or F, X 5 is D or H, X 5 is N, D, A or E (SEQ ID NO: 129); (v) a HVR-L2 comprising the amino acid sequence WAX 1 TLAX 2 , wherein X 1 is S or E, X 2 is S, Y, F or W (SEQ ID NO: 130); and (vi) a HVR-L3 comprising the amino acid sequence AGGYGGGX 1 YA, wherein X 1 is L or R (SEQ ID NO: 131); or (d) (i) a HVR-L1 comprising the amino acid sequence X 1 X 2 SQX 3 VX 4 X 5 X 6 NWLS, wherein X 1 is Q or T, X 2 is S or T, X 3 is S or E, X 4 is Y or F, X 5 is D or H, X 6 is N, D, A or E (SEQ ID NO: 129); (ii) a HVR-L2 comprising the amino acid sequence WAX 1 TLAX 2 , wherein X 1 is S or E, X 2 is S, Y, F or W (SEQ ID NO: 130); and (iii) a HVR-L3 comprising the amino acid sequence AGGYGGGGX 1 YA, wherein X 1 is L or R (SEQ ID NO: 131).

7 . The antibody of claim 6 , wherein,
(a) the antibody of 6(b), further comprises a heavy chain variable domain framework FR1 comprising the amino acid sequence of SEQ ID NO: 132, 133, or 134; FR2 comprising the amino acid sequence of SEQ ID NO: 135 or 136; FR3 comprising the amino acid sequence of SEQ ID NO: 137; and FR4 comprising the amino acid sequence of SEQ ID NO: 138; or (b) the antibody of claim 6 (d), further comprises a light chain variable domain framework FR1 comprising the amino acid sequence of SEQ ID NO: 139; FR2 comprising the amino acid sequence of SEQ ID NO: 140 or 141; FR3 comprising the amino acid sequence of SEQ ID NO: 142 or 143; and FR4 comprising the amino acid sequence of SEQ ID NO: 144.

8 . The antibody of claim 1 , comprising (a) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 13, 16-30, 32-34, 86-94, or 95; (b) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 15, 31, 35-38, 96-98, or 99; or (c) a VH sequence of SEQ ID NO: 13, 16-30, 32-34, 86-94, or 95, and a VL sequence of SEQ ID NO: 15, 31, 35-38, 96-98, or 99.

9 . The antibody of claim 8 , further comprising a variant Fc region amino acid sequence of SEQ ID NO: 229-380, or 381.

10 . An isolated nucleic acid encoding the antibody of claim 1 .

11 . A host cell comprising the nucleic acid of claim 10 .

12 . A method of producing an anti-myostatin antibody comprising:
(a) culturing the host cell of claim 11 so that the antibody is produced; or (b) immunizing an animal against a polypeptide, wherein the polypeptide comprises the region corresponding to amino acids at positions 21 to 100 of myostatin propeptide (SEQ ID NO: 78).

13 . A method of detecting the presence of latent myostatin or myostatin propeptide comprising
(a) contacting a biological sample with the antibody of claim 2 under conditions permissive for binding of the antibody to latent myostatin or myostatin propeptide, and (b) detecting whether a complex is formed between the antibody and the latent myostatin or myostatin propeptide.

14 . A method of blocking release of mature myostatin from latent myostatin in a biological sample comprising contacting a biological sample containing latent myostatin with the antibody of claim 2 under conditions permissive for binding of the antibody to the latent myostatin, and blocking release of mature myostatin from the latent myostatin.

15 . A method of reducing the concentration of latent myostatin in a biological sample comprising contacting a biological sample containing latent myostatin with the antibody of claim 2 under conditions permissive for binding of the antibody to the latent myostatin, and removing the complex formed between the antibody and the latent myostatin.

16 . A pharmaceutical formulation comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.

17 . A method of treating an individual having a muscle wasting disease comprising administering to the individual an effective amount of the antibody of claim 1 .

18 . A method of increasing mass of muscle tissue in an individual comprising administering to the individual an effective amount of the antibody of claim 1 to increase mass of muscle tissue.

19 . A method of increasing strength of muscle tissue in an individual comprising administering to the individual an effective amount of the antibody of claim 1 to increase strength of muscle tissue.

20 . A method of reducing body fat accumulation in an individual comprising administering to the individual an effective amount of the antibody of claim 1 to reduce body fat accumulation.

21 . A polypeptide comprising a variant Fc region comprising at least one amino acid alteration in a parent Fc region, wherein the ratio of [KD value of the parent Fc region for monkey FcγRIIb]/[KD value of the variant Fc region for monkey FcγRIIb] is 2.0 or great

Assignees

• Chugai Pharmaceutical Co Ltd

Inventors

• Ruike Yoshinao
• Kuramochi Taichi
• Muramatsu Hiroyasu
• Ueyama Atsunori
• Igawa Tomoyuki
• Katada Hitoshi
• Hori Yuji

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P9/04

  Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title