# US 10,864,268 B2 · CD47 antibodies, methods, and uses

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- Number: US 10,864,268 B2
- Document ID: US10864268B2
- Status: Granted
- Kind: B2
- Application number: 15/527,551
- Filed: 2015-11-17
- Published: 2018-09-06
- Granted: 2020-12-15
- Earliest claimed: 2014-11-18
- Assignees: Janssen Pharmaceutica NV
- Applicants: Janssen Pharmaceutica NV
- Inventors: Christine Pietsch; Jianying Dong; Rosa Cardoso; Hong Zhou
- Web page: https://patentgrep.com/patent/US10864268B2

## CPC

- A61K 39/39558: {against tumor tissues, cells, antigens}
- C07K 16/2803: {against the immunoglobulin superfamily}
- C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere}
- A61K 2039/505: {comprising antibodies}
- A61P 35/00: Antineoplastic agents
- C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2317/55: Fab or Fab'
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/567: Framework region [FR]
- C07K 2317/71: Decreased effector function due to an Fc-modification
- C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K 2317/734: Complement-dependent cytotoxicity [CDC]
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present disclosure relates generally to monoclonal antibodies that specifically bind to CD47, more specifically to CD47 antibodies that do not have significant platelet aggregation activity and do not have significant hemagglutination activity. Methods of generating these antibodies and methods of using these monoclonal antibodies as therapeutics are also provided.

## Claims

### Claim 1

An isolated antibody or antigen binding fragment thereof comprising the following properties:
a. the antibody or antigen binding fragment specifically binds to CD47 by interacting with CD47 (SEQ ID NO: 21 excluding the signal sequence) amino acid residues:
i. Q1, L3, N27, E29, E97, L101, T102, R103, and E104;
ii. Y37, K39, R45, D46, T49, A53, L54, N55, K56, S57, T58, V59, P60, T61, T564, A66, and K67;
iii. E35, K39, Y37, D46, 49, D51, A53, L54, K56, T58, V59, T99, L101, and T102; or
iv. E29, Q31, N32, E35, V36, Y37, K39, N41, D46, D51, A53, E97, T99, E100, L101, T102, R103, and E104;
b. the antibody or antigen binding fragment thereof prevents CD47 from interacting with signal-regulatory protein alpha (SIRP alpha);
c. the antibody or antigen binding fragment thereof does not have significant platelet aggregation activity and
wherein the antibody or antigen binding fragment thereof comprises a VH complementarity determining region 1 (CDR1) sequence set forth in SEQ ID NO: 9 or SEQ ID NO: 10, a VH CDR2 sequence set forth in SEQ ID NO: 11 or SEQ ID NO: 12, a VH CDR3 sequence set forth in SEQ ID NO: 13 or SEQ ID NO: 14, a VL CDR1 sequence set forth in SEQ ID NO: 15 or SEQ ID NO: 16, a VL CDR2 sequence set forth in SEQ ID NO: 17 or SEQ ID NO: 18 and a VL CDR3 sequence set forth in SEQ ID NO: 19 or SEQ ID NO: 20.

### Claim 2 (depends on 1)

The antibody of claim 1 wherein the antibody or antigen binding fragment specifically binds to human or cyno CD47, and comprises a variable heavy (VH) chain region selected from the group consisting of SEQ ID NOs: 4-6.

### Claim 3 (depends on 2)

The antibody of claim 2, wherein the antibody or antigen binding fragment thereof comprises a variable light (VL) chain region selected from the group consisting of SEQ ID NOs: 7 and 8.

### Claim 4 (depends on 2)

The antibody of claim 2, wherein the antibody or antigen binding fragment thereof comprises a VH region selected from the group consisting of SEQ ID NOs: 4-6 and a VL region selected from the group consisting of SEQ ID NOs: 7 and 8.

### Claim 5 (depends on 4)

The antibody of claim 4, wherein the VH chain region comprises SEQ ID NO: 4 or SEQ ID NO: 5 paired with a VL chain region comprising SEQ ID NO: 7.

### Claim 6 (depends on 4)

The antibody of claim 4, wherein the VH chain region comprises SEQ ID NO: 6 paired with a VL chain region comprising SEQ ID NO: 8.

### Claim 7 (depends on 1)

The antibody of claim 1, wherein the antibody or antigen binding fragment thereof comprises a VH CDR1 sequence set forth in SEQ ID NO: 9, a VH CDR2 sequence set forth in SEQ ID NO: 11, a VH CDR3 sequence set forth in SEQ ID NO: 13, a VL CDR1 sequence set forth in SEQ ID NO: 15, a VL CDR2 sequence set forth in SEQ ID NO: 17, and a VL CDR3 sequence set forth in SEQ ID NO: 19.

### Claim 8 (depends on 1)

The antibody of claim 1, wherein the antibody or antigen binding fragment thereof comprises a VH CDR1 sequence set forth in SEQ ID NO: 10, a VH CDR2 sequence set forth in SEQ ID NO: 12, a VH CDR3 sequence set forth in SEQ ID NO: 14, a VL CDR1 set forth in SEQ ID NO: 16, a VL CDR2 sequence set forth in SEQ ID NO: 18, and a VL CDR3 sequence set forth in SEQ ID NO: 20.

### Claim 9 (depends on 2)

The antibody of claim 2, wherein the VH chain of the antibody has more extensive contacts with CD47 than the VL chain of the antibody.

### Claim 10 (depends on 2)

The antibody of claim 2, wherein the epitope bound by the VH chain of the antibody is positioned near the membrane of a CD47 expressing cell, and wherein the VL chain of the antibody occludes a SIRP alpha binding site on CD47.

### Claim 11 (depends on 2)

The antibody of claim 2, wherein the antibody does not have significant hemagglutination activity.

### Claim 12 (depends on 2)

The antibody of claim 2, wherein the platelet-aggregation activity of the antibody is no more than 10% greater than the degree of platelet-aggregation observed in the absence of the antibody.

### Claim 13 (depends on 2)

The antibody of claim 2, wherein the antibody is chimeric, humanized, or fully human.

### Claim 14 (depends on 2)

The antibody of claim 2, wherein the CD47 is human CD47 or cyno CD47.

### Claim 15 (depends on 2)

The antibody of claim 2, wherein the antibody or antigen binding fragment thereof promotes macrophage-mediated phagocytosis of a CD47-expressing cell.

### Claim 16 (depends on 2)

The antibody of claim 2, wherein the antibody or antigen binding fragment thereof comprises an IgG isotype selected from the group consisting of IgG1 isotype and IgG2 isotype.

### Claim 17 (depends on 2)

A pharmaceutical composition comprising the antibody of claim 2 and a pharmaceutical acceptable carrier.