# US 12,215,131 B2 · IgG Fc-IL2-Rα-IL2 fusions and methods of use thereof

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- Number: US 12,215,131 B2
- Document ID: US12215131B2
- Status: Granted
- Kind: B2
- Application number: 18/314,205
- Filed: 2023-05-09
- Published: 2023-10-19
- Granted: 2025-02-04
- Earliest claimed: 2019-12-20
- Assignees: Regeneron Pharmaceuticals, Inc.
- Applicants: Regeneron Pharmaceuticals, Inc.
- Inventors: Jiaxi Wu; Tong Zhang; Maria del Pilar Molina-Portela; Eric Smith; Chia-Yang Lin; Thomas Craig Meagher
- Web page: https://patentgrep.com/patent/US12215131B2

## CPC

- C07K 14/55: IL-2
- A61K 35/17: Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00)
- A61K 38/2013: {IL-2}
- A61K 39/0011: {Cancer antigens}
- A61K 39/12: Viral antigens
- A61K 39/39: characterised by the immunostimulating additives, e.g. chemical adjuvants
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/31: Chimeric antigen receptors [CAR]
- A61K 40/4234: {Interleukins [IL]}
- A61K 40/46: Viral antigens
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- A61P 31/14: for RNA viruses
- A61P 31/20: for DNA viruses
- A61P 35/00: Antineoplastic agents
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- C07K 14/5443: {IL-15}
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/70517: {CD8}
- C07K 14/70539: {MHC-molecules, e.g. HLA-molecules}
- C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)}
- C07K 14/7155: {for interleukins [IL]}
- C07K 16/2818: {against CD28 or CD152}
- 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}
- A61K 2039/585: {wherein the target is cancer}
- A61K 2239/50: Colon
- A61K 2239/55: Lung
- A61K 2239/57: Skin; melanoma
- C07K 16/084: Papillomaviridae (F); Polyomaviridae (F), e.g. SV40, BK virus or JC virus
- 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 2317/622: Single chain antibody (scFv)
- C07K 2319/00: Fusion polypeptide
- C07K 2319/02: containing a signal sequence
- C07K 2319/03: containing a transmembrane segment
- 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
- C12N 2710/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2710/20034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2740/13034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- 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

The present disclosure relates to IL2 agonists with improved therapeutic profiles.

## Claims

### Claim 1

A fusion protein comprising a polypeptide chain which comprises, in an N- to C-terminal orientation:
(a) a variant IgG1 Fc domain or a variant IgG4 Fc domain comprising one or more amino acid substitutions that reduce binding to an Fc receptor and/or reduce effector function; and
(b) an IL2 moiety comprising, in an N- to C-terminal orientation:
(i) an IL2-Rα domain comprising the amino acid sequence of SEQ ID NO:4; and
(ii) an IL2 domain comprising the amino acid sequence of SEQ ID NO:2.

### Claim 2 (depends on 1)

The fusion protein of claim 1, wherein the variant IgG1 Fc domain or variant IgG4 Fc domain has reduced binding to an Fcγ receptor.

### Claim 3 (depends on 2)

The fusion protein of claim 2, wherein the Fcγ receptor is human FcγRIIIa, FcγRI or FcγRIIa.

### Claim 4 (depends on 1)

The fusion protein of claim 1, wherein the polypeptide chain comprises a variant IgG1 Fc domain.

### Claim 5 (depends on 1)

The fusion protein of claim 1, wherein the variant IgG1 Fc domain or variant IgG4 Fc domain has reduced complement dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), or cytokine secretion.

### Claim 6 (depends on 1)

The fusion protein of claim 1, wherein the polypeptide chain comprises a variant IgG4 Fc domain.

### Claim 7 (depends on 4)

The fusion protein of claim 4, wherein the variant IgG1 Fc domain comprises the amino acid mutations L234A and L235A, numbered according to Kabat EU index.

### Claim 8 (depends on 1)

The fusion protein of claim 1, wherein the variant IgG1 Fc domain or variant IgG4 Fc domain comprises a hinge region.

### Claim 9 (depends on 8)

The fusion protein of claim 8, wherein the hinge region is a modified hinge region that reduces binding affinity for an Fcγ receptor relative to a wild-type hinge region of the same isotype.

### Claim 10 (depends on 8)

The fusion protein of claim 8, wherein positions 233-236 within the hinge region are G, G, G and unoccupied; G, G, unoccupied, and unoccupied; G, unoccupied, unoccupied, and unoccupied; or all unoccupied, with positions numbered by EU numbering.

### Claim 11 (depends on 9)

The fusion protein of claim 9, wherein the modified hinge region comprises the amino acid sequence CPPCPAPGGG-GPSVF (SEQ ID NO:82), CPPCPAPGG-GPSVF (SEQ ID NO:83), CPPCPAPG-GPSVF (SEQ ID NO:84), or CPPCPAP-GPSVF (SEQ ID NO:85).

### Claim 12 (depends on 1)

The fusion protein of claim 1, wherein the IL2-Rα domain and the IL2 domain are separated by a linker.

### Claim 13 (depends on 12)

The fusion protein of claim 12, wherein the linker separating the IL2 domain and the IL2-Rα domain is 2 to 60 amino acids in length.

### Claim 14 (depends on 1)

The fusion protein of claim 1, wherein the variant IgG1 Fc domain or variant IgG4 Fc domain and the IL2 moiety are separated by a linker.

### Claim 15 (depends on 14)

The fusion protein of claim 14, wherein the linker separating the variant IgG1 Fc domain or variant IgG4 Fc domain and the IL2 moiety is 2 to 60 amino acids in length.

### Claim 16 (depends on 1)

The fusion protein of claim 1, which is a dimer.

### Claim 17 (depends on 16)

The fusion protein of claim 16, which comprises a second polypeptide chain, said second polypeptide chain comprising in an N- to C-terminal orientation:
(a) a variant IgG1 Fc domain comprising one or more amino acid substitutions that reduce binding to an Fc receptor and/or reduce effector function; and
(b) an IL2 moiety comprising, in an N- to C-terminal orientation:
(i) an IL2-Rα domain comprising the amino acid sequence of SEQ ID NO:4; and
(ii) an IL2 domain comprising the amino acid sequence of SEQ ID NO:2 and separated from the IL2-Rα domain only by a noncleavable linker.

### Claim 18 (depends on 16)

The fusion protein of claim 16, which is a homodimer.

### Claim 19 (depends on 1)

A nucleic acid or plurality of nucleic acids encoding the fusion protein of claim 1.

### Claim 20 (depends on 1)

A host cell engineered to express the fusion protein of claim 1.

### Claim 21 (depends on 17)

The fusion protein of claim 17, which is a homodimer.

### Claim 22 (depends on 16)

The fusion protein of claim 16, which comprises a second polypeptide chain, said second polypeptide chain comprising in an N- to C-terminal orientation:
(a) a variant IgG4 Fc domain comprising one or more amino acid substitutions that reduce binding to an Fc receptor and/or reduce effector function; and
(b) an IL2 moiety comprising, in an N- to C-terminal orientation:
(i) an IL2-Rα domain comprising the amino acid sequence of SEQ ID NO:4; and
(ii) an IL2 domain comprising the amino acid sequence of SEQ ID NO:2 and separated from the IL2-Rα domain only by a noncleavable linker.

### Claim 23 (depends on 22)

The fusion protein of claim 22, which is a homodimer.

### Claim 24 (depends on 23)

A pharmaceutical composition comprising the fusion protein of claim 23 and an excipient.

### Claim 25 (depends on 23)

A method of treating skin, breast, lung, or colon cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the fusion protein of claim 23.