# US 2020/0399344 A1 · Nk cell-directed chimeric proteins

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- Number: US 2020/0399344 A1
- Document ID: US20200399344A1
- Status: Published application
- Kind: A1
- Application number: 17/017,117
- Filed: 2020-09-10
- Published: 2020-12-24
- Earliest claimed: 2019-05-16
- Assignees: Shattuck Labs, Inc.
- Applicants: Shattuck Labs, Inc.
- Inventors: Taylor SCHREIBER; Casey SHUPTRINE; George FROMM; Suresh DE SILVA
- Web page: https://patentgrep.com/patent/US20200399344A1

## CPC

- C07K 14/70532: {B7 molecules, e.g. CD80, CD86}
- C07K 14/7056: {Lectin superfamily, e.g. CD23, CD72}
- 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 47/68: the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61P 35/00: Antineoplastic agents
- C07K 14/70503: {Immunoglobulin superfamily}
- C07K 14/70539: {MHC-molecules, e.g. HLA-molecules}
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C07K 2319/70: containing domain for protein-protein interaction
- 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 present invention relates, inter alia, to compositions and methods, including chimeric proteins comprising an extracellular domain of a Type I transmembrane protein or a portion of a membrane-anchored extracellular protein and a portion of the extracellular domain of a Type II transmembrane protein, wherein the Type II transmembrane protein is naturally expressed on the surface of a Natural Killer (NK) cell that find use in the treatment of disease, such as cancer and viral infections.

## Claims

### Claim at position 1

-93. (canceled)

### Claim 94

A chimeric protein of a general structure of:
N terminus-(a)-(b)-(c)-C terminus,
wherein:
(a) is a first domain comprising an extracellular domain of CD86, the first domain capable of binding a CD86 ligand/receptor,
(b) is a linker, and
(c) is a second domain comprising an extracellular domain of NKG2A, the second domain capable of binding an NKG2A ligand/receptor,
wherein the linker connects the first domain and the second domain.

### Claim 95 (depends on 94)

The chimeric protein of claim 94, wherein the NKG2A ligand/receptor is HLA-E.

### Claim 96 (depends on 94)

The chimeric protein of claim 94, wherein binding the NKG2A ligand/receptor blocks transmission of an immune inhibitory signal to an NK cell.

### Claim 97 (depends on 94)

The chimeric protein of claim 94, wherein the CD86 ligand/receptor is selected from CTLA-4 and CD28.

### Claim 98 (depends on 94)

The chimeric protein of claim 94, wherein the linker is a polypeptide selected from a flexible amino acid sequence, an IgG hinge region, and an antibody sequence.

### Claim 99 (depends on 94)

The chimeric protein of claim 94, wherein the linker comprises at least one cysteine residue capable of forming a disulfide bond and/or comprises a hinge-CH2-CH3 Fc domain.

### Claim 100 (depends on 99)

The chimeric protein of claim 99, wherein the hinge-CH2-CH3 Fc domain is derived from IgG, IgA, IgD, or IgE, optionally wherein the IgG is selected from IgG1, IgG2, IgG3, and IgG4, and the IgA is selected from IgA1 and IgA2.

### Claim 101 (depends on 100)

The chimeric protein of claim 100, wherein the IgG is IgG4.

### Claim 102 (depends on 101)

The chimeric protein of claim 101, wherein the IgG4 is a human IgG4.

### Claim 103 (depends on 94)

The chimeric protein of claim 94, wherein the linker comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

### Claim 104 (depends on 103)

The chimeric protein of claim 103, wherein the linker further comprises one or more joining linkers, such joining linkers being independently selected from SEQ ID NOs: 4-50.

### Claim 105 (depends on 94)

The chimeric protein of claim 94, wherein the first domain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 63.

### Claim 106 (depends on 94)

The chimeric protein of claim 94, wherein the second domain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 57.

### Claim 107 (depends on 94)

The chimeric protein of claim 94, wherein the chimeric protein has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 65.

### Claim 108 (depends on 94)

The chimeric protein of claim 94, wherein binding of the first domain to its ligand/receptor inhibits an immunosuppressive signal and/or activates another immunosuppressive signal.

### Claim 109 (depends on 94)

The chimeric protein of claim 94, wherein the chimeric protein is capable of forming a stable synapse between an NK cell and a tumor cell, or an NK cell and a virus-infected cell.

### Claim 110 (depends on 109)

The chimeric protein of claim 109, wherein the stable synapse between cells provides spatial orientation that favors tumor reduction or killing of virus-infected cells by the NK cell.

### Claim 111 (depends on 110)

The chimeric protein of claim 110, wherein the spatial orientation positions NK cells to attack target cells selected from tumor cells and virus-infected cells and/or sterically prevents the target cells from delivering signals, including immune inhibiting signals beyond those masked by the chimeric protein of the invention.

### Claim 112 (depends on 94)

The chimeric protein of claim 94, wherein binding of either or both of the first domain and the second domain to its ligand/receptor occurs with slow off rates (Koff), which provides a long interaction of a receptor and its ligand.

### Claim 113 (depends on 112)

The chimeric protein of claim 112, wherein the long interaction provides sustained negative signal masking effect, sustained inhibition of an immunosuppressive signal, and/or sustained activation of an immunosuppressive signal.

### Claim 114 (depends on 112)

The chimeric protein of claim 112, wherein the long interaction provides for NK cell proliferation and/or allows for anti-tumor attack or attack of a virus-infected cell.

### Claim 115 (depends on 112)

The chimeric protein of claim 112, wherein the long interaction allows sufficient signal transmission to provide release of a stimulatory signal.

### Claim 116 (depends on 115)

The chimeric protein of claim 115, wherein the stimulatory signal is a cytokine.

### Claim 117

A chimeric protein comprising:
(a) a first domain comprising a portion of CD86 comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 63,
(b) a second domain comprising a portion of NKG2A comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 57, and
(c) a linker linking the first domain and the second domain and comprising a hinge-CH2-CH3 Fc domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, and optionally further comprising one, two, or more joining linkers independently selected from SEQ ID NOs: 4-50.

### Claim 118 (depends on 117)

The chimeric protein of claim 117, wherein the chimeric protein comprises an amino acid sequence having at least about 95% sequence identity with the amino acid sequence of SEQ ID NO: 65.

### Claim 119 (depends on 94)

A pharmaceutical composition comprising the chimeric protein of claim 94.

### Claim 120 (depends on 119)

A method of treating cancer or an inflammatory disease, comprising administering an effective amount of a pharmaceutical composition of claim 119 to a subject in need thereof.