Cytokine-based therapies and methods
The disclosure relates to methods for redirecting an active form of an endogenous cytokine to a target cell or target tissue of interest in a biological system or subject in need thereof by administering a multi-specific binding molecule comprising (a) a binding domain that specifically binds to an active form of a cytokine and (b) a binding domain that specifically binds to an epitope on a molecule that is a marker on a target cell or tissue, wherein the multi-specific binding molecule, when bound to the cytokine, does not block or only partially blocks, the ability of the cytokine to bind to and agonize a cognate receptor for the cytokine.
1 . A method for developing a multi-specific binding molecule that redirects an active form of a cytokine to a target cell or a target tissue of interest, the method comprising:
(a) providing a panel that comprises a plurality of cytokine-binding domains that bind to the cytokine and a plurality of target-binding domains that bind to a target molecule, wherein the target molecule is a marker on the target cell in the target tissue;
(b) screening the panel using an assay that measures the ability of the cytokine, when complexed with at least one cytokine-binding domain, to bind to and/or agonize its cognate receptor compared to the ability of an unbound cytokine to bind to and/or agonize its cognate receptor;
(c) screening the plurality of target-binding domains for binding to the target molecule;
(d) selecting at least one cytokine-binding domain among the panel that does not block or only partially blocks the ability of the cytokine to bind to and/or agonize its cognate receptor;
(e) generating a panel of multi-specific binding molecules, wherein the panel of multispecific binding molecules comprise the at least one cytokine-binding domain selected in (d) and at least one target-binding domain selected in (c); and
(f) screening the panel of multi-specific binding molecules in an in vitro cell-based assay that measures the ability of the cytokine to bind to and agonize its cognate receptor in the presence of varying amounts of the multi-specific binding molecules.
2 . The method of claim 1 , further comprising screening the panel of multi-specific binding molecules in an in vivo assay in a non-human subject that measures the ability of the cytokine to bind to and agonize its cognate receptor when administered to the subject.
3 . The method of claim 1 , further comprising performing an epitope binning assay in conjunction with the screening of the panel of cytokine-binding domains to identify a region or regions on the cytokine that, when bound to the cytokine-binding domain, retain or partially retain the ability of the cytokine to bind to and agonize its cognate receptor.
4 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules comprises:
(a) one cytokine-binding domain and one target-binding domain;
(b) one cytokine-binding domain and two target-binding domains;
(c) two cytokine-binding domains and one target-binding domain; or
(d) two identical or non-identical cytokine-binding domains and two identical or non-identical target-binding domains.
5 . The method of claim 1 , wherein the multi-specific binding molecule comprises a cytokine-binding domain that specifically binds to a human interleukin.
6 . The method of claim 5 , wherein the human interleukin is human: IL-1, IL-2, IL-4, IL-5, Il-6, IL-8 IL-12, IL-18, or IL-23.
7 . The method of claim 1 , wherein the multi-specific binding molecule comprises a cytokine-binding domain that specifically binds to IL-15 or to IL-15 complexed with IL-15 receptor alpha.
8 . The method of claim 7 , wherein the cytokine-binding domain binds to an IL-15 or to IL-15-IL:15 receptor alpha complex with an affinity of less than 100 nM as measured by surface plasmon resonance (SPR).
9 . The method of claim 7 , wherein the cytokine-binding domain binds to an IL-15 or to IL-15-IL:15 receptor alpha complex with an affinity of less than 0.1 nM as measured by surface plasmon resonance (SPR).
10 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules comprises a target-binding domain that specifically binds to a protein expressed in a tumor microenvironment (TME).
11 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules comprises a target-binding domain that specifically binds to a tumor-associated antigen (TAA) expressed on the surface of a tumor cell, and wherein the cytokine is redirected to the location of the tumor cell.
12 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules comprises a target-binding domain that specifically binds to a receptor on an immune cell.
13 . The method of claim 12 , wherein the immune cell is a T cell.
14 . The method of claim 12 , wherein the immune cell is a macrophage.
15 . The method of claim 12 , wherein the immune cell is an NK-cell.
16 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules comprises a cytokine-binding domain and a target-binding domain that bind a cognate cytokine receptor and a target that are on the same cell.
17 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules comprises a cytokine-binding domain and a target-binding domain that bind to a cognate cytokine receptor and a target that are on different cells.
18 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules comprises a scaffold.
19 . The method of claim 18 , wherein the scaffold is an albumin-based scaffold, a fibronectin-based scaffold or an immunoglobin-based scaffold.
20 . The method of claim 19 , wherein the scaffold is an immunoglobulin-based scaffold, and wherein the immunoglobulin-based scaffold is derived from an IgG1, an IgG2, an IgG4, an IgM, or an IgA.
21 . The method of claim 19 , wherein the scaffold is an albumin-based scaffold or an immunoglobulin-based scaffold, and wherein the albumin-based or the immunoglobulin-based scaffold is capable of binding to a neonatal Fc receptor (FcRn).
22 . The method of claim 1 , wherein a multi-specific binding molecule of the panel of multi-specific binding molecules is a bi-specific antibody comprising a binding domain that specifically binds to an epitope on the active form of the cytokine and a binding domain that specifically binds to an epitope on a molecule that is not a cytokine receptor on the target cell or tissue.
23 . The method of claim 1 , wherein the multi-specific binding molecule treats a disease when administered to a subject in need thereof.