# US 2023/0265178 A1 · Modified immunoglobulins for targeting amyloid deposits

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- Number: US 2023/0265178 A1
- Document ID: US20230265178A1
- Status: Published application
- Kind: A1
- Application number: 18/181,489
- Filed: 2023-03-09
- Published: 2023-08-24
- Earliest claimed: 2019-11-15
- Assignees: Attralus, Inc.; University of Tennessee Research Foundation
- Applicants: Attralus, Inc.; University of Tennessee Research Foundation
- Inventors: Jonathan S. WALL; James S. FOSTER; Spencer GUTHRIE
- Web page: https://patentgrep.com/patent/US20230265178A1

## CPC

- C07K 16/18: against material from animals or humans
- A61K 47/6843: {the antibody targeting a material from animals or humans}
- A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
- C07K 14/4711: {Alzheimer's disease; Amyloid plaque core protein}
- C12N 15/85: for animal cells
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- A61K 2039/505: {comprising antibodies}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2317/55: Fab or Fab'
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance
- C07K 2319/00: Fusion polypeptide
- C07K 2319/01: containing a localisation/targetting motif
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
- 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)
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided herein are modified immunoglobulins comprising an amyloid reactive peptide joined to an antibody, as well as humanized antibodies that bind to human amyloid fibrils and antibody-peptide fusion proteins. Also provided herein are methods of treating amyloid-based diseases by administering a modified immunoglobulin, humanized antibody, or antibody-peptide fusion protein.

## Claims

### Claim at position 1

1-100. (canceled)

### Claim 101

A modified immunoglobulin, comprising:
(i) an amyloid reactive peptide; and
(ii) an Ig antibody or functional fragment thereof that binds to human amyloid fibrils, wherein the Ig antibody or functional fragment thereof comprises a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), wherein:
a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19;
b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19; or
c) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.

### Claim 102 (depends on 101)

The modified immunoglobulin of claim 101, wherein the amyloid reactive peptide comprises an amino acid sequence having at least 85% sequence identity to any one of the amino acid sequences set forth in SEQ ID NOs:1-14.

### Claim 103 (depends on 101)

The modified immunoglobulin of claim 101, wherein the amyloid reactive peptide comprises an amino acid sequence having at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2.

### Claim 104 (depends on 101)

The modified immunoglobulin of claim 101, wherein the Ig antibody or functional fragment comprises human framework sequences.

### Claim 105 (depends on 101)

The modified immunoglobulin of claim 101, wherein the Ig antibody or functional fragment thereof comprises a human Fc region.

### Claim 106 (depends on 101)

The modified immunoglobulin of claim 101, wherein the modified immunoglobulin binds to rVλ6Wil fibrils, Per125 wtATTR extract, KEN hATTR extract, SHI ALλ liver extract, TAL ALκ liver extract, Aβ, Aβ(1-40), IAAP, ALκ4, A1λ1, or ATTR fibrils.

### Claim 107 (depends on 101)

The modified immunoglobulin of claim 101, wherein the modified immunoglobulin binds to human amyloid fibrils with a KD that is less than about 1000 nM.

### Claim 108 (depends on 101)

The modified immunoglobulin of claim 101, wherein the modified immunoglobulin has an EC50 that is less than 100 nM.

### Claim 109 (depends on 101)

The modified immunoglobulin of claim 101, wherein the modified immunoglobulin is conjugated to a detectable label.

### Claim 110 (depends on 101)

The modified immunoglobulin of claim 101, wherein the IgG antibody is humanized.

### Claim 111 (depends on 101)

The modified immunoglobulin of claim 101, wherein the VL comprises one or more amino acid residues selected from the group consisting of:
a. Tyr at position 36;
b. Leu at position 37;
c. Leu at position 46;
d. Leu at position 85; and
e. Phe at position 87,
wherein the amino acid positions are numbered according to the numbering system of Kabat.

### Claim 112 (depends on 101)

The modified immunoglobulin of claim 101,
wherein the VH comprises one or more amino acid residues selected from the group consisting of:
a. Val at position 37;
b. Leu at position 48;
c. Leu at position 67;
d. Ser at position 68;
e. Lys at position 71;
f. Ser at position 76;
g. Val at position 78;
h. Leu at position 79;
i. Phe at position 80;
j. Thr at position 89;
k. Val at position 93; and
l. Thr at position 94
wherein the amino acid positions are numbered according to the numbering system of Kabat.

### Claim 113 (depends on 112)

The modified immunoglobulin of claim 112, wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises Leu at position 48, Ser at position 76, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94.

### Claim 114 (depends on 101)

The modified immunoglobulin of claim 101, wherein the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32-42 and/or the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63.

### Claim 115 (depends on 101)

The modified immunoglobulin of claim 101, wherein the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32-42 and/or the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63.

### Claim 116 (depends on 101)

The modified immunoglobulin of claim 101, wherein the VL comprises the amino acid sequence set forth in of SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 55.

### Claim 117 (depends on 101)

The modified immunoglobulin of claim 101, wherein the
the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.

### Claim 118 (depends on 101)

A pharmaceutical composition comprising the modified immunoglobulin of claim 101.

### Claim 119 (depends on 101)

Nucleic acid(s) encoding the modified immunoglobulin of claim 101.

### Claim 120 (depends on 119)

A host cell comprising the nucleic acid(s) of claim 119.

### Claim 121 (depends on 120)

A method of making a modified immunoglobulin comprising culturing the host cell of claim 120 under conditions suitable for expression of the vector encoding the modified immunoglobulin and recovering the modified immunoglobulin.

### Claim 122 (depends on 101)

A method of treating a subject having an amyloid related disorder, comprising administering to the subject an effective amount of the modified immunoglobulin of claim 101.

### Claim 123 (depends on 101)

A method of identifying an amyloid deposit in a subject, comprising:
administering the modified immunoglobulin of claim 101 to the subject; and
detecting a signal from the modified immunoglobulin,
wherein the modified immunoglobulin comprises a detectable label.

### Claim 124

A humanized antibody that binds to human amyloid fibrils, wherein the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein
a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19;
b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19; or
c) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.

### Claim 125 (depends on 124)

A pharmaceutical composition comprising the humanized antibody of claim 124.

### Claim 126 (depends on 124)

Nucleic acid(s) encoding the humanized antibody of claim 124.

### Claim 127 (depends on 126)

A host cell comprising the nucleic acids(s) of claim 126.

### Claim 128 (depends on 127)

A method of making a humanized antibody comprising culturing the host cell of claim 127 under conditions suitable for expression of the vector encoding the humanized antibody and recovering the humanized antibody.

### Claim 129 (depends on 124)

A method of treating a subject having an amyloid related disorder, comprising administering to the subject an effective amount of the humanized antibody of claim 124.

### Claim 130 (depends on 124)

A method of identifying an amyloid deposit in a subject, comprising detectably labeling the humanized antibody of claim 124, administering the humanized antibody to the subject, and detecting a signal from the humanized antibody.