Multispecific NK engager protein

Patent metadata

Field	Value
Publication number	US-12252543-B2
Application number	US-201816173150-A
Country	US
Kind code	B2
Filing date	Oct 29, 2018
Priority date	Jun 23, 2015
Publication date	Mar 18, 2025
Grant date	Mar 18, 2025

Abstract

Multispecific proteins that bind and specifically redirect NK cells to lyse a target cell of interest are provided without non-specific activation of NK cells in absence of target cells. The proteins have utility in the treatment of disease, notably cancer or infectious disease.

First claim

The invention claimed is:

1. A nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein comprising:
(a) a first antigen binding domain (ABD) which:
(i) monovalently binds to a human NKp46 polypeptide having the amino acid sequence of SEQ ID NO: 1; and
(ii) comprises a first immunoglobulin heavy chain variable region (VH1) and a first immunoglobulin light chain variable region (VL1);

(b) a second ABD which:
(i) binds to a first antigen of interest other than an internalizing cytokine receptor alpha chain or B-cell maturation antigen (BCMA); and
(ii) comprises a second immunoglobulin heavy chain variable region (VH2) and a second immunoglobulin light chain variable region (VL2); and

(c) a dimeric human Fc polypeptide capable of binding to CD16A,

or a vector or vectors containing the nucleic acid or nucleic acids.

2. An isolated cell which comprises a nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein according to claim 1 .

3. A method of producing a multispecific antigen binding protein comprising culturing an isolated cell according to claim 2 .

4. A nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein or a vector or vectors containing the nucleic acid or nucleic acids according to claim 1 , wherein the first antigen of interest is expressed by a solid tumor.

5. A nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein or a vector or vectors containing the nucleic acid or nucleic acids according to claim 1 , wherein the first antigen of interest is expressed by a hematological cancer.

6. A nucleic acid or nucleic acids encoding a multispecific antigen binding protein or a vector or vectors containing the nucleic acid or nucleic acids according to claim 1 , wherein the antigen of interest is expressed by an infectious agent.

7. A nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein or a vector or vectors containing the nucleic acid or nucleic acids according to claim 1 , wherein:
(i) the dimeric human Fc polypeptide is modified to enhance CD16A binding relative to the corresponding wild-type Fc polypeptide;
(ii) the dimeric human Fc polypeptide is capable of binding to human neonatal Fc receptor (FcRn);
(iii) the multispecific protein binds to the first antigen of interest monovalently;
(iv) the multispecific antigen binding protein mediates lysis of target cells by a combination of NKp46 signaling and CD16A-mediated antibody-dependent cell-mediated cytotoxicity (ADCC);
(v) the multispecific antigen binding protein mediates NKp46-signaling-mediated lysis and CD16-mediated lysis resulting in a synergistic enhancement in the lysis of target cells;
(vi) the multispecific antigen binding protein mediates lysis of target cells by a combination of NKp46-signaling, CD137-signaling and CD16A-mediated ADCC;
(vii) the second ABD comprises a monovalent or bivalent antibody or antibody fragment comprising the VH2 and the VL2;
(viii) (ix) the spacing between the first ABD and the second ABD is less than 80 angstroms;
(ix) the spacing between the first ABD and the second ABD is about 55 angstroms;
(x) the multispecific antigen binding protein is a heterodimeric, or heterotrimeric protein;
(xi) the multispecific antigen binding protein comprises at least one polypeptide chain that comprises an ABD bound to the dimeric human Fc polypeptide by a flexible linker;
(xii) the multispecific antigen binding protein comprises at least one polypeptide chain that comprises an ABD bound to the dimeric human Fc polypeptide by a flexible polypeptide linker;
(xiii)
(xiii-1) the multispecific antigen binding protein comprises: a first polypeptide comprising an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL) separated by a polypeptide linker; and a second polypeptide comprising a VH and a VL separated by a polypeptide linker;
(xiii-2) the multispecific antigen binding protein comprises:
a first polypeptide comprising the VH1 and the VL1 separated by a polypeptide linker; and a second polypeptide comprising the VH2 and the VL2 separated by a polypeptide linker;

(xiii-3) the multispecific antigen binding protein comprises: a first polypeptide comprising the VH1 and the VL2; and a second polypeptide comprising the VH2 and the VL1;
(xiii-4) the first ABD comprises a Fab comprising the VH1 and the VL1 or comprises the VH1 and the VL1 separated by a linker, optionally by a polypeptide linker;
(xiii-5) the second ABD comprises a Fab comprising the VH2 and the VL2 or comprises the VH2 and VL2 separated by a linker, optionally by a linker comprising a linear or cyclic peptide;
(xiii-6) the first ABD comprises a Fab comprising the VH1 and the VL1 or comprises the VH1 and the VL1 separated by a linker; and the second ABD comprises a Fab comprising the VH2 and the VL2 or comprises the VH2 and the VL2 separated by a linker;
(xiii-7) the first ABD comprises a Fab comprising the VH1 and the VL1; and the second ABD comprises the VH2 and the VL2 separated by a linker; or
(xiii-8) the first ABD comprises the VH1 and the VL1 separated by a linker; and the second ABD comprises a Fab comprising the VH2 and the VL2;
(xiv) the multispecific antigen binding protein further comprises a third ABD which binds to a second antigen of interest, wherein the second antigen of interest is same as or different from the first antigen of interest and is different from said human NKp46 polypeptide;
(xv) the first antigen of interest comprises a cell surface polypeptide which is, when bound by a full-length IgG1 antibody,
(xv-1) capable of undergoing intracellular internalization or
(xv-2) not capable of undergoing intracellular internalization;
(xvi) the multispecific protein does not substantially increase or induce intracellular internalization of the first antigen of interest on target cells;
(xvii) the multispecific antigen binding protein upregulates CD137 expression on NK cells;
(xviii) the multispecific antigen binding protein elicits the expression of CD137 on the surface of NKp46 + CD16 + NK cells; or
(xix) any combination of the foregoing.

8. A nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein or a vector or vectors containing the nucleic acid or nucleic acids according to claim 1 , wherein the dimeric human Fc polypeptide is capable of binding to human FcRn.

9. A nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein or a vector or vectors containing the nucleic acid or nucleic acids according to claim 8 , wherein the dimeric human Fc polypeptide comprises one or more amino acid modifications that increase binding affinity for CD16A.

10. A nucleic acid which encodes or nucleic acids which separately or in combination encode a multispecific antigen binding protein or a vector or vectors containing the nucleic acid or nucleic acids according to claim 1 , wherein the multispecific antigen binding protein comprises a first (central) polypeptide chain comprising one variable domain that, together with a complementary variable domain comprised in a second polypeptide chain, forms an antigen binding domain specific for one antigen of interest, and the first (central) polypeptide chain further comprises a second variable domain that is paired with a complementary variable domain

Assignees

• Innate Pharma

Inventors

• Gauthier Laurent
• Anceriz Nadia
• Morel Ariane
• Rossi Benjamin

Classifications

• C07K2317/75

  Agonist effect on antigen · CPC title

• C07K2317/732

  Antibody-dependent cellular cytotoxicity [ADCC] · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• C07K2317/526

  CH3 domain · CPC title