MULTISPECIFIC NKp46 BINDING PROTEINS

Patent metadata

Field	Value
Publication number	US-2024141042-A1
Application number	US-202318490804-A
Country	US
Kind code	A1
Filing date	Oct 20, 2023
Priority date	Jun 27, 2014
Publication date	May 2, 2024
Grant date	—

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 infections disease.

First claim

What is claimed is:

1 . An isolated multispecific protein comprising a first antigen binding domain and a second antigen binding domain, wherein one of the first or second antigen binding domains binds to a human NKp46 polypeptide and the other binds an antigen of interest, wherein the multispecific protein binds the NKp46 polypeptide monovalently, and wherein the multispecific protein is capable of directing an NKp46-expressing NK cell to lyse a target cell expressing the antigen of interest.

2 - 49 . (canceled)

50 . The multispecific protein according to claim 1 , wherein
(i) said lysis of the target cell is mediated by NKp46-signaling; (ii) the multispecific protein does not exhibit activation of NKp46-expressing NK cells when incubated with such NK cells in the absence of cells expressing the antigen of interest; (iii) the multispecific protein does not exhibit activation of NKp46-negative, CD16-positive lymphocytes when incubated with such NK cells in the presence of cells expressing the antigen of interest; (iv) the multispecific protein (a) activates NK cells, when incubated with NKp46-expressing NK cells and target cells; and (b) does not activate NKp46-expressing NK cells when incubated with NK cells in the absence of target cells; (v) the multispecific protein does not exhibit activation of NKp46-expressing NK cells when incubated with NK cells and target cells, in the presence of Fcγ-expressing cells; (vi) the human NKp46 polypeptide is a polypeptide comprising the amino acid sequence of SEQ ID NO: 1; (vii) the protein comprises at least a portion of an Fc domain, is capable of binding to human neonatal Fc receptor (FcRn) and has decreased binding to a human Fcγ receptor compared to a full length wild type human IgG1 antibody, optionally wherein the Fc domain is interposed between the two antigen binding domains; (viii) the multispecific protein competes for binding to a NKp46 polypeptide with any one or any combination of monoclonal antibodies NKp46-1, NKp46-2, NKp46-3, NKp46-4, NKp46-6 or NKp46-9, or the Anti-CD19-IgG1-F2-NKp46-1, -2, -3, -4, -6 or -9 bispecific antibodies; (ix) the multispecific protein has decreased binding to a mutant NKp46 polypeptide selected from the group consisting of:
a. a mutant NKp46 polypeptide having a mutation at residues R101, V102, E104 and/or L105 compared to binding to the wild-type NKp46;
b. a mutant NKp46 polypeptide having a mutation any one or more of the residues K41, E42, μl 19, Y121 and/or Y194 compared to binding to the wild-type NKp46; and
c. a mutant NKp46 polypeptide having a mutation any one or more of the residues P132, E133, I135, and/or S136 compared to binding to the wild-type NKp46;
(x) the multispecific protein substantially lacks binding to a human Fcγ receptor; (xi) the multispecific protein is a single chain protein comprising:
(a) a first antigen binding domain that binds to NKp46;
(b) a second antigen binding domain that binds a polypeptide expressed on a target cell; and
(c) at least a portion of a human Fc domain, wherein the multispecific polypeptide is capable of binding to human neonatal Fc receptor (FcRn) and has decreased binding to a human Fcγ receptor compared to a full length wild type human IgG1 antibody;
(xii) the Fc domain comprises
(a) a CH2 domain, and
(b) a CH3 domain with a modification, optionally an amino acid mutation, to prevent CH3-CH3 dimerization.
(xiii) the CH3 domain comprises an amino acid substitution at 1, 2, 3, 4, 5, 6 or 7 of the positions L351, T366, L368, P395, F405, T407 and/or K409 (EU numbering as in Kabat); (xiv) the Fc domain comprises (i) a CH2 domain, and (ii) a first and a second CH3 domain separated by a linker peptide, wherein the two CH3 domains associate with one another via non-covalent interactions; (xv) the multispecific protein comprises an Fc domain interposed between the first antigen binding domain and the second binding domain; (xvi) the multispecific protein comprises an Fe domain interposed between the first antigen binding domain and the second binding domain, wherein the protein comprises a polypeptide having a domain arrangement: (ABD1)-CH2-CH3-(ABD2); (xvii) the multispecific protein comprises an Fc domain interposed between the first antigen binding domain and the second binding domain, wherein the protein comprises a polypeptide having a domain arrangement: (ABD1)-linker-CH2-CH3-linker-(ABD2); (xviii) the multispecific protein comprises an Fc domain interposed between the first antigen binding domain and the second binding domain, wherein the protein comprises a polypeptide having a domain arrangement: (ABD1)-linker-CH2-CH3-linker-CH3-linker-(ABD2); or (xix) any combination of the foregoing.

51 . The multispecific protein according to claim 1 , wherein the multispecific protein is an isolated heterodimeric polypeptide comprising:
(1) an isolated heterodimeric polypeptide comprising:
(a) a first polypeptide chain comprising, from N- to C-terminus, a first variable domain (V), a CH1 of CK constant region, a Fc domain or portion thereof, a second variable domain and a third variable domain; and
(b) a second polypeptide chain comprising, from N- to C-terminus, a first variable domain (V), a CH1 or CK constant region, and optionally a Fc domain or portion thereof, wherein the CH1 or CK constant region is selected to be complementary to the CH1 or CK constant region of the first polypeptide chain such that the first and second polypeptides form a CH1-CK heterodimer in which the first variable domain of the first polypeptide chain and the first variable domain of the second polypeptide form an antigen binding domain that binds the first antigen of interest; and wherein a second variable domain and third variable domain forms an antigen binding domain that binds the second antigen of interest; or
(2) an isolated heterodimeric polypeptide comprising:
(a) a first polypeptide having a domain arrangement selected from:
Va-1-(CH1 or CK)a-Fc domain-Va-2-Vb-2, and
Va-2-Vb-2-Fc domain-Va-1-(CH1 or CK)b,
and
(b) a second polypeptide chain having a domain arrangement:
Vb-1-(CH1 or CK)b, and
wherein one of Va-1 and Vb-1 is a light chain variable domain and the other is a heavy chain variable domain, one of Va-2 and Vb-2 is a light chain variable domain and the other is a heavy chain variable domain;
wherein (CH1 or CK)b dimerizes with the (CH1 or CK)a on the central chain, and the Vb-1 forms an antigen binding domain together with Va-1 of the central chain, and wherein Va-2 and Vb-2 together form an antigen binding domain; or
(3) an isolated heterodimeric polypeptide comprising: a first polypeptide having a domain arrangement:
V a-1 -(CH1 or CK) a -Fc domain-V a-2 -V b-2 ,
and
(b) a second polypeptide chain having a domain arrangement:
V b-1 -(CH1 or CK) b -Fc domain

wherein one of Va-1 and Vb-1 is a light chain variable domain and the other is a heavy chain variable domain, one of Va-2 and Vb-2 is a light chain variable domain and the other is a heavy chain variable domain; said (CH1 or CK)b dimerizes with the (CH1 or CK)a on the central chain, and the Vb-1 forms an antigen binding domain together with Va-1 of the central chain; and Va-2 and Vb-2 together form an antigen binding domain.

52 . The multispecific protein according to claim 51 , wherein Va-2 and Vb-2 together form an antigen binding domain that binds NKp46.

53 . The multispecific protein according to claim 1 , comprising:
(1) an isolated heterotrimeric polypeptide comprising
(a) a first polypeptide chain comprising, from N- to C-terminus, a first variable domain (V) fused to a first CH1 or CK constant region, an Fc domain or portion thereof, and a second variable domain (V) fused to a second CH1 or CK constant region;
(b) a second polypeptide c

Assignees

• Innate Pharma

Inventors

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

Classifications

• C07K16/2803 Primary

  against the immunoglobulin superfamily · CPC title

• C07K16/2866

  against receptors for cytokines, lymphokines, interferons · CPC title

• C07K2317/31

  multispecific · CPC title

• C07K2317/71

  Decreased effector function due to an Fc-modification · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title