# US 11,993,657 B2 · Bifunctional molecules for treatment of immune disorders

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,993,657 B2
- Document ID: US11993657B2
- Status: Granted
- Kind: B2
- Application number: 17/694,570
- Filed: 2022-03-14
- Published: 2022-09-29
- Granted: 2024-05-28
- Earliest claimed: 2021-03-16
- Assignees: JN Biosciences LLC
- Applicants: Jn Biosciences LLC
- Inventors: Naoya Tsurushita; J. Yun Tso
- Web page: https://patentgrep.com/patent/US11993657B2

## CPC

- C07K 16/2866: {against receptors for cytokines, lymphokines, interferons}
- A61K 47/6811: {the drug being a protein or peptide, e.g. transferrin or bleomycin}
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- A61K 47/6889: {Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment}
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- C07K 14/70521: {CD28, CD152}
- C07K 14/7155: {for interleukins [IL]}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/31: multispecific
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2319/00: Fusion polypeptide
- 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

## Abstract

The invention provides bifunctional molecules including an antibody specifically binding to CD122 and an extracellular domain of CTLA-4. The bifunctional molecules specifically bind to CD122 and CTLA-4 ligands, CD80 and CD86 and inhibit their function in immune activation. The bifunctional molecules can inhibit interaction of CD122 with its ligands IL-2 and IL-15 and inhibit interaction of CD80 and CD86 with their counter-receptor, CD28. These bifunctional molecules can suppress Signals 2 and 3 of immune responses as a single therapeutic agent for treatment of immune disorders.

## Claims

### Claim 1

A bifunctional molecule comprising an antibody that binds to CD122 and inhibits its interaction with IL-2 and IL-15, comprising
(a) a mature light chain linked to a CTLA-4 extracellular domain as a chain comprising SEQ ID NO:34 or 42 and a mature heavy chain comprising SEQ ID NO:8 or 33 provided the C-terminal lysine can be omitted, or
(b) a mature heavy chain linked to a CTLA-4 extracellular domain as a chain comprising SEQ ID NO:39 provided the C-terminal lysine can be omitted, and a mature light chain comprising SEQ ID NO:9.

### Claim 2 (depends on 1)

The bifunctional molecule of claim 1, in the form of a heterodimer comprising two copies of the heavy chain and two copies of the light chain.

### Claim 3

A bifunctional molecule comprising a mature heavy chain linked to a CTLA-4 extracellular domain as a chain comprising SEQ ID NO:13 or 16 and a mature light chain comprising SEQ ID NO:9.

### Claim 4 (depends on 1)

The bifunctional molecule of claim 1, wherein the mature light chain is linked to the CTLA-4 extracellular domain as the chain comprising SEQ ID NO:34 and the mature heavy chain comprises SEQ ID NO:8 or 33 provided the C-terminal lysine can be omitted.

### Claim 5 (depends on 1)

The bifunctional molecule of claim 1, wherein the mature heavy chain is linked to the CTLA-4 extracellular domain as the chain comprising SEQ ID NO:39 provided the C-terminal lysine can be omitted, and the mature light chain comprises SEQ ID NO:9.

### Claim 6 (depends on 1)

The bifunctional molecule of claim 1, wherein the mature light chain is linked to the CTLA-4 extracellular domain as the chain comprising SEQ ID NO:42, and the mature heavy chain comprises SEQ ID NO:8 or 33 provided the C-terminal lysine can be omitted.

### Claim 7

A bifunctional molecule comprising a mature heavy chain linked to a CTLA-4 extracellular domain as a chain comprising SEQ ID NO:57 and a mature light chain comprising SEQ ID NO:54.

### Claim 8

A bifunctional molecule comprising a mature heavy chain linked to a CLTA-4 extracellular domain as a chain comprising SEQ ID NO:56 provided the C-terminal lysine can be omitted, and the mature light chain comprises SEQ ID NO:54.

### Claim 9

A bifunctional molecule comprising an antibody that binds to CD122 and inhibits its interaction with IL-2 and IL-15, wherein the antibody comprises a heavy chain comprising a mature heavy chain variable region linked to a heavy chain constant region, and a light chain comprising a mature light chain variable region linked to a light chain constant region, wherein the antibody is linked to a CTLA-4 extracellular domain having at least 90% sequence identity with SEQ ID NO:10 and the mature heavy chain variable region comprises three heavy chain CDRs comprising SEQ ID NOS:59-61 respectively and the mature light chain variable region comprises three light chain CDRs comprising SEQ ID NOS:63-65 respectively.

### Claim 10 (depends on 9)

The bifunctional molecule of claim 9, wherein the mature heavy chain variable region comprises SEQ ID NO:58 and the mature light chain variable region comprises SEQ ID NO:62.

### Claim 11 (depends on 9)

The bifunctional molecule of claim 9, wherein the mature heavy chain comprises SEQ ID NO:66 provided the C-terminal lysine can be omitted, and the mature light chain comprises SEQ ID NO:67.

### Claim 12 (depends on 9)

The bifunctional molecule of claim 9, wherein the antibody is linked to the extracellular domain of CTLA-4 via a polypeptide linker.

### Claim 13 (depends on 12)

The bifunctional molecule of claim 12, wherein the polypeptide linker comprises the amino acid sequence of SEQ ID NO:11, 36 or 50.

### Claim 14

A bifunctional molecule comprising a mature heavy chain linked to a CTLA-4 extracellular domain as a chain comprising SEQ ID NO:71 and a mature light chain comprising SEQ ID NO:67.

### Claim 15 (depends on 9)

The bifunctional molecule of claim 9, wherein the mature heavy chain is linked to the CTLA-4 extracellular domain as a chain comprising SEQ ID NO:70 provided the C-terminal lysine can be omitted, and the mature light chain comprises SEQ ID NO:67.

### Claim 16 (depends on 9)

The bifunctional molecule of claim 9, wherein the antibody has human IgG1 kappa isotype.

### Claim 17 (depends on 9)

The bifunctional molecule of claim 9, wherein the heavy chain constant region has one or more mutations to reduce effector function.

### Claim 18 (depends on 17)

The bifunctional molecule of claim 17, wherein the mutations are L234A and L235A.

### Claim 19 (depends on 9)

The bifunctional molecule of claim 9, wherein the extracellular domain of CTLA-4 comprises SEQ ID NO:10.

### Claim 20 (depends on 9)

The bifunctional molecule of claim 9, wherein the extracellular domain of CTLA-4 comprises SEQ ID NO:14.

### Claim 21 (depends on 1)

A pharmaceutical composition comprising a bifunctional molecule of claim 1 and a pharmaceutically acceptable carrier.

### Claim 22 (depends on 1)

A method of treating a subject having an autoimmune disease, comprising administering an effective regime of the bifunctional molecule of claim 1 to the subject.

### Claim 23 (depends on 22)

The method of claim 22, wherein the autoimmune disease is any of vitiligo, alopecia areata, type 1 diabetes, celiac disease, multiple sclerosis, polymyositis, allergic dermatitis, primary biliary cirrhosis, Behcet's disease, and ulcerative colitis.

### Claim 24 (depends on 1)

A method of treating a subject having non-alcoholic steatohepatitis, comprising administering an effective regime of the bifunctional molecule of claim 1 to the subject.

### Claim 25 (depends on 1)

A method of treating a subject having skin allograft rejection, comprising administering an effective regime of the bifunctional molecule of claim 1 to the subject.