Antagonistic interleukin-6 chimeric proteins

Abstract

The present invention relates, in part, to chimeric proteins, protein complexes, and Fc-based chimeric protein complexes comprising an IL-6 antagonist and their use as therapeutic agents. The present invention further relates to methods of treatment of various diseases, such as autoimmune disease or disorder.

Description

[0001] ORN-109PC / 114384-5109

[0002] ANTAGONISTIC INTERLEUKIN-6 CHIMERIC PROTEINS

[0003] FIELD

[0004] The present invention relates, in part, to chimeric proteins, or chimeric protein complexes (including Fc-based chimeric protein complexes) comprising an interleukin-6 (IL-6) antagonist and their use as therapeutic agents

[0005] CROSS-REFERENCE TO RELATED APPLICATIONS

[0006] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 703,304, filed October 4, 2024, and U.S Provisional Application No. 63 / 848,649, filed July 22, 2025, the entire contents of which are hereby incorporated by reference in their entireties.

[0007] SEQUENCE LISTING

[0008] The contents of the computer readable Sequence Listing in XM L format (“XML Document”) submitted electronically herewith are incorporated herein by reference in their entirety. A computer readable format copy of the Sequence Listing (filename: ORN-109PC_114384-5109.xml, date produced: September 21 , 2025; size: 589,306 bytes) is submitted per 37 C.F.R. §§ 1.831-1.835.

[0009] BACKGROUND

[0010] Interleukins are mostly secreted proteins that act as cytokines to mediate inter-cellular communication during immune responses. Interleukin-6 (IL-6) is a 184 amino acid polypeptide belonging to the class of helical cytokines. IL-6, which was originally identified as a B-cell differentiation factor, is now known to be a multifunctional cytokine that regulates the immune response, hematopoiesis, the acute phase response, and inflammation. Furthermore, IL-6 is an important regulator of the acute phase response, T cell function, and terminal B cell differentiation. Deregulation of IL-6 production is implicated in the pathology of several disease processes. For example, excessive or inappropriate production of this cytokine may be involved in a variety of autoimmune and neoplastic disorders.

[0011] Since its discovery more than 40 years ago, the IL-6 pathway has emerged as a pivotal pathway involved in immune regulation in health and dysregulation in many diseases. Targeting the IL-6 pathway may provide various innovative therapeutic approaches for a variety of disorders and diseases.

[0012] Accordingly, there remains a need for safe and effective IL-6 antagonist-based therapeutics with improved target selectivity, pharmacokinetic and therapeutic properties and minimal toxicity profiles.

[0013] SUMMARY

[0014] In some aspects, the present invention relates to chimeric proteins and chimeric protein complexes, including Fc- based chimeric protein complexes, comprising an interleukin-6 (IL-6) antagonist to inhibit IL-6 signaling. In some embodiments, the IL-6 antagonist is a modified wild type IL-6, wherein the wild type IL-6 has the amino acid sequence of SEQ ID NO: 1 In some embodiments, the IL-6 antagonist has one or more substitution mutations at a position selected from Y31, G35, L57, E59, N60, S118, V121, and W157, where the residue numbering is based

[0015] 1

[0016] DB1 / 162518159.2 ORN-109PC / 114384-5109 on SEQ ID NO: 426 (mature IL-6 after signal peptide cleavage after position 28). In some embodiments, the one or more of these mutations produce a modified human IL-6 with ablated activity and / or affinity for IL-6 receptor (gp130).

[0017] In some aspects, the present invention provides a chimeric protein comprising: (a) an IL-6 antagonist, and (b) one or more targeting moieties, said targeting moieties comprising recognition domains which specifically bind to an antigen or receptor of interest; wherein the IL-6 antagonist and the one or more targeting moieties are optionally connected with one or more linkers.

[0018] In another aspect, the present invention is related to a Fc-based chimeric protein complex comprising: a) an IL-6 antagonist, and b) one or more targeting moieties, said targeting moieties comprising recognition domains which specifically bind to an antigen or receptor of interest; and c) a Fc domain, the Fc domain optionally having one or more mutations that reduces or eliminates one or more effector functions of the Fc domain, promotes Fc chain pairing in the Fc domain, and / or stabilizes a hinge region in the Fc domain.

[0019] The present invention concerns a target-specific IL-6 antagonist having loss-of-function site II and / or site III mutations that confer reduced or ablated activity and / or affinity for the IL-6 receptor (gp130). In emobodiments, the antagonistic activity of IL-6 still binds strongly when attached to a targeting moiety, said targeting moiety comprising a recognition domain which specifically binds to an antigen or receptor of interest. In embodiments, the target-specific IL-6 antagonist comprises one or more site II and / or site III mutations that inhibit recruitment of two gp 130 receptor chains to the I L-6 / I L-6Ra complex and higher order clustering.

[0020] The present disclosure concerns, in part, findings that chimeric proteins or chimeric protein complexes (including Fc-based chimeric protein complexes) comprising an IL-6 antagonist, exhibit substantially targeted IL-6R (IL-6 Receptor) inhibition upon IL-6 induced activation. This targeted IL-6R-inhibition can be induced at a target cell when directed to such a cell through a targeting moiety. Surprisingly, the induced IL-6 inhibition activity at a target cell, achieved through targeting of chimeric proteins or chimeric protein complexes comprising an IL-6 antagonist may inhibit an excess of IL-6 (e.g., inhibit 3000 pg / mL of IL-6 with an IL-6 antagonist IC50 at 10 pg / ml). Importantly, the IL-6 antagonist chimeric proteins and chimeric protein complexes comprising IL-6 antagonist described herein, exhibit substantial and surprising selectivity for target cells as compared to non-target cells In summary, a unique combination of highly potent and highly cell target-selective IL-6R inhibition can be achieved with IL-6 antagonist compositions described herein.

[0021] In some embodiments, the IL-6 antagonist is a modified wild-type IL-6, i.e., is a variant and comprises one or more mutations in IL-6. In some embodiments, the one or more mutations reduce or ablate the biological activity of the IL-6 (sometimes referred to as "loss-of-function’’ mutation), creating an IL-6 antagonist. For example, the one or more mutations may inhibit IL-6 signaling. In an embodiment, the IL-6 antagonist comprises one or more mutations that ablate its affinity and / or activity for IL-6R (gp130). In some embodiments, the IL-6 inhibition activity can be induced upon directing or targeting of the chimeric protein or chimeric protein complex comprising the IL-6 antagonist to a target cell through a targeting moiety.

[0022] DB1 / 162518159.2 2 ORN-109PC / 114384-5109

[0023] In various embodiments, the chimeric proteins and chimeric protein complexes, including Fc-based chimeric protein complexes, comprise one or more targeting moieties which have recognition domains (e.g. antigen recognition domains, including without limitation various antibody formats, inclusive of single-domain antibodies) which specifically bind to a target (e.g. antigen, receptor) of interest. In various embodiments, the targeting moieties have recognition domains that specifically bind to a target (e.g. antigen, receptor) of interest, including those found on one or more immune cells, which can include, without limitation, T cells, cytotoxic T lymphocytes, T helper cells, T regulatory cells (Tregs), natural killer (NK) cells, natural killer T (NKT) cells, anti-tumor and tumor macrophages (e.g. M1 and M2 macrophages), B cells, B regulatory (Breg) cells, neutrophils, monocytes, myeloid derived cells, adipose stromal cells (ASC) and dendritic cells. In various embodiments, the targeting moieties have recognition domains that specifically bind to a target (e.g. antigen, receptor) of interest, including those found on one or more tumor cells, endothelial cells, epithelial cells, mesenchymal cells, stromal cells or other cell types that are characteristic of and / or unique for specific organs and / or tissues, including those specifically associated with disease. In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) of interest and effectively recruit one or more immune cells. In some embodiments, the targets (e.g. antigens, receptors) of interest can be found on one or more tumor cells. In some embodiments, the present chimeric proteins, chimeric protein complexes, including Fc-based chimeric protein complexes, may recruit an immune cell, e.g., an immune cell that can kill and / or suppress a tumor cell, or modulate other immune cells, to a site of action (such as, by way of non-limiting example, the tumor microenvironment). In some embodiments, the present chimeric proteins, chimeric protein complexes, including Fc-based chimeric protein complexes, may modulate an immune cell at a site of action, or recruit an immune cell to a site of action that is associated with an autoimmune disease, inflammatory disease, infection, metabolic and / or cardiovascular disease (such as, by way of non-limiting example, the disease microenvironment) In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) of interest that is part of a non-cellular structure.

[0024] In some embodiments, the chimeric proteins or the chimeric protein complexes described herein comprise IL-6 antagonists (which is an example of a cytokine / signaling agent inhibitor that may also be used in the present invention) with reduced and / or ablated biological activity that is coupled to one or more targeting moieties. In some embodiments, the chimeric proteins or chimeric protein complexes of the present invention include AnTakines (Antagonistic activity-on-Target cytokines) that have one or more mutated cytokines that have their antagonistic and / or inhibitive activity delivered upon target cell binding. In some embodiments, without limitation, the chimeric proteins or protein complexes or chimeric protein complexes target an IL-6 antagonist to hepatocytes, adipose stromal cells (ASC), NK cells, CD4 T cells, and CD8 T cells. In vivo, such chimeric proteins or chimeric protein complexes can potently promote the response of hepatocytes, adipose stromal cells (ASC), NK cells, CD4 T cells, and CD8 T cells to antigens, with a significantly reduced toxicity profile compared to wild-type (WT) IL-6.

[0025] In some embodiments, the targeting moiety binds to one or more of the follow targets: CD8, CTLA4, CD3, CD4, DNAM-1, Nrp1 (neurophilin), ASC, TNFR2, GITR, ICOS, CD20, CD70, Clec9a, NKp46, asialoglycoprotein receptor

[0026] DB1 / 162518159.2 3 ORN-109PC / 114384-5109

[0027] (ASGPR), PD-1 , PD-L1, PD-L2, SIRP1 a, FAP, XCR1, tenascin, prostate-specific membrane antigen (PSMA), and ECM proteins.

[0028] In various embodiments, the present chimeric proteins and chimeric protein complexes, including Fc-based chimeric protein complexes find use in the treatment of various diseases or disorders such as autoimmune diseases or disorders, immune disorders, cancer, hepatocellular carcinoma, pancreatic cancer / melanoma, obesity, Graft-versus-host disease (GVHD), viral infections, cardiovascular diseases, wound, ischemia-related diseases, neurodegenerative diseases, and / or metabolic diseases and many other diseases and disorders, and the present invention encompasses various methods of treatment.

[0029] Another aspect of the invention is related to a method for treating a subject afflicted with an autoimmune disease or disorder, comprising administering a chimeric protein or chimeric protein complex as described herein. In some embodiments, the chimeric protein or chimeric protein complex comprises: (i) an IL-6 antagonist, (ii) one or more targeting moieties, said targeting moieties comprising recognition domains which specifically bind to an antigen or receptor of interest; and (iii) a connector between (i) and (ii), the connector being: (1) an Fc domain, the Fc domain optionally having one or more mutations that reduces or eliminates one or more effector functions of the Fc domain, promotes Fc chain pairing in the Fc domain, and / or stabilizes a hinge region in the Fc domain that connects (i) and (ii) and / or (2) a flexible linker that connects (i) and (ii); wherein the IL-6 antagonist is characterized by reduced or ablated affinity and / or activity (e g., inhibition activity) at the IL-6 receptor (gp130).

[0030] Importantly, the chimeric proteins and chimeric protein complexes comprising an IL-6 antagonist described herein, exhibit substantial and surprising selectivity for target cells versus non-target cells. In summary, a unique combination of highly potent and highly cell target-selective signaling inhibition can be achieved with the chimeric proteins or chimeric protein complexes described herein.

[0031] In embodiments, the chimeric proteins and chimeric protein complexes comprising an IL-6 antagonist described herein comprises a polypeptide having an amino acid sequence selected from SEQ ID NO: 414-425, 439-456.

[0032] BRIEF DESCRIPTION OF THE DRAWINGS

[0033] FIGs. 1A-F, 2A-H, 3A-H, 4A-D, 5A-F, 6A-J, 7A-D, 8A-F, 9A-J, 10A-F, 11A-L, 12A-L, 13A-F, 14A-L, 15A-L, 16A- J, 17A-J, 18A-F, and 19A-F show various non-limiting illustrative schematics of the Fc-based chimeric protein complexes of the present invention. In embodiments, each schematic is a composition of the present invention. Where applicable in the figures, ”TM" refers to a “targeting moiety” as described herein, “SA” refers to a “signaling agent” as described herein, “*“l” is an optional “linker" as described herein, the two long parallel rectangles are human Fc domains, e.g. from lgG1 , from lgG2, or from lgG4, as described herein and optionally with effector knock-out and / or stabilization mutations as also described herein, and the two long parallel rectangles with one having a protrusion and the other having an indentation are human Fc domains, e.g. from I gG 1 , from lgG2, or from lgG4 as described herein, with knob-in-hole and / or ionic pair (a / k / a charged pairs, ionic bond, or charged residue pair) mutations as described herein and optionally with effector knock-out and / or stabilization mutations as also described herein.

[0034] DB1 / 162518159.2 4 ORN-109PC / 114384-5109

[0035] FIGs. 1A-F show illustrative homodimeric 2-chain complexes. These figures show illustrative configurations for the homodimeric 2-chain complexes.

[0036] FIGs. 2A-H show illustrative homodimeric 2-chain complexes with two targeting moieties (TM) (as described herein, more targeting moieties may be present in some embodiments). In embodiments, the position of TM1 and TM2 are interchangeable. In embodiments, the constructs shown in the box ( / .e., Figs. 2G and 2H) have signaling agent (SA) between TM1 and TM2 or between TM1 and Fc.

[0037] FIGs. 3A-H show illustrative homodimeric 2-chain complexes with two signaling agents (as described herein, more signaling agents may be present in some embodiments). In embodiments, the position of SA1 and SA2 are interchangeable. In embodiments, the constructs shown in the box (ie., Figs. 3G and 3H) have TM between SA1 and SA2 or TM at N- or C-terminus.

[0038] FIGs. 4A-D show illustrative heterodimeric 2-chain complexes with split TM and SA chains, namely the TM on the knob chain of the Fc and the SA on hole chain of the Fc.

[0039] FIGs. 5A-F show illustrative heterodimeric 2-chain complexes with split TM and SA chains, namely with both TMs on the knob chain of the Fc and with SA on hole chain of the Fc, with two targeting moieties (as described herein, more targeting moieties may be present in some embodiments) In embodiments, the position of TM1 and TM2 are interchangeable. In some embodiments, TM1 and TM2 can be identical.

[0040] FIGs. 6A-J show illustrative heterodimeric 2-chain complexes with split TM and SA chains, namely with TM on the knob chain of the Fc and with a SA on the hole chain of the Fc, with two signaling agents (as described herein, more signaling agents may be present in some embodiments). In these orientations and / or configurations, one SA is on the knob chain and one SA is on the hole chain. In embodiments, the position of SA1 and SA2 are interchangeable.

[0041] FIGs. 7A-D show illustrative heterodimeric 2-chain complexes with split TM and SA chains, namely the SA on the knob chain of the Fc and the TM on hole chain of the Fc.

[0042] FIGs. 8A-F show illustrative heterodimeric 2-chain complexes with split TM and SA chains, namely with SA on the knob chain of the Fc and both TMs on hole chain of the Fc, with two targeting moieties (as described herein, more targeting moieties may be present in some embodiments). In embodiments, the position of TM1 and TM2 are interchangeable. In some embodiments, TM1 and TM2 can be identical.

[0043] FIGs. 9A-J show illustrative heterodimeric 2-chain complexes with split TM and SA chains, namely with SA on the knob chain of the Fc and TM on hole chain of the Fc, with two signaling agents (as described herein, more signaling agents may be present in some embodiments). In these orientations and / or configurations, one SA is on the knob chain and one SA is on the hole chain. In embodiments, the position of SA1 and SA2 are interchangeable.

[0044] FIGs. 10A-F show illustrative heterodimeric 2-chain complexes with TM and SA on the same chain, namely the SA and TM both on the knob chain of the Fc.

[0045] DB1 / 162518159.2 5 ORN-109PC / 114384-5109

[0046] FIGs. 11A-L show illustrative heterodimeric 2-chain complexes with a TM and a SA on the same chain, namely with SA and with TM both on the knob chain of the Fc, with two targeting moieties (as described herein, more targeting moieties may be present in some embodiments). In embodiments, the position of TM1 and TM2 are interchangeable. In some embodiments, TM1 and TM2 can be identical.

[0047] FIGs. 12A-L show illustrative heterodimeric 2-chain complexes with a TM and a SA on the same chain, namely with SA and with TM both on the knob chain of the Fc, with two signaling agents (as described herein, more signaling agents may be present in some embodiments). In embodiments, the position of SA1 and SA2 are interchangeable.

[0048] FIGs. 13A-F show illustrative heterodimeric 2-chain complexes with TM and SA on the same chain, namely the SA and TM both on the hole chain of the Fc

[0049] FIGs. 14A-L show illustrative heterodimeric 2-chain complexes with a TM and a SA on the same chain, namely with SA and with TM both on the hole chain of the Fc, with two targeting moieties (as described herein, more targeting moieties are present in some embodiments). In embodiments, the position of TM1 and TM2 are interchangeable. In embodiments, TM1 and TM2 can be identical.

[0050] FIGs. 15A-L show illustrative heterodimeric 2-chain complexes with a TM and a SA on the same chain, namely with SA and with TM both on the hole chain of the Fc, with two signaling agents (as described herein, more signaling agents may be present in some embodiments). In embodiments, the position of SA1 and SA2 are interchangeable.

[0051] FIGs. 16A-J show illustrative heterodimeric 2-chain complexes with two targeting moieties (as described herein, more targeting moieties may be present in some embodiments) and with SA on knob Fc and TM on each chain. In embodiments, TM1 and TM2 can be identical.

[0052] FIGs. 17A-J show illustrative heterodimeric 2-chain complexes with two targeting moieties (as described herein, more targeting moieties may be present in some embodiments) and with SA on hole Fc and TM on each chain. In embodiments, TM1 and TM2 can be identical.

[0053] FIGs. 18A-F show illustrative heterodimeric 2-chain complexes with two signaling agents (as described herein, more signaling agents may be present in some embodiments) and with split SA and TM chains: SA on knob and TM on hole Fc.

[0054] FIGs. 19A-F show illustrative heterodimeric 2-chain complexes with two signaling agents (as described herein, more signaling agents may be present in some embodiments) and with split SA and TM chains: TM on knob and SA on hole Fc.

[0055] FIG. 20 shows STAT3 activation by CD20-IL-6 wild-type or the different CD20-IL-6 site II and / or site III mutants on MCF7-CD20 cells

[0056] FIG. 21 depicts inhibition of STAT3 activation by Belli 0 / CD20-IL-6 Y31 D / G35F + L57D / E59F / N60W in the presence of saturating IL-6 concentrations on MCF7 and MCF7-CD20 cells.

[0057] DB1 / 162518159.2 6 ORN-109PC / 114384-5109

[0058] FIG. 22 shows inhibition of STAT3 activation by CD20-IL-6 Y31D / G35F + L57D / E59F / N60W in the presence of different IL-6 concentrations (1 , 4, and 16 ng / ml) on MCF7-CD20 cells.

[0059] FIG. 23 depicts STAT3 activation by CD20-targeted IL-6 site II + / - site III mutants on MCF7 and MCF7-CD20 cells.

[0060] FIG. 24 shows inhibition of STAT3 activation by Bcll10 / CD20-l L-6 Y31G L57A W157A / G in the presence of saturating IL-6 concentrations on MCF7 and MCF7-CD20 cells.

[0061] FIG. 25 depicts inhibition of STAT3 activation by CD20-AnTa6 variants in the presence of different IL-6 concentrations (3 and 12 ng / ml) on MCF7-CD20 cells.

[0062] FIG. 26 shows sensorgrams of gp130 binding to the preformed complex of IL-6Ra with IL-6 or IL-6 antagonist variants.

[0063] FIG. 27 depicts schematic representations of the Fc-based ASGPR-IL-6 antagonist construct (left) and the untargeted IL-6 antagonist control (right).

[0064] FIG. 28 shows the inhibitory capacity of the Fc-based ASGPR VHH-IL-6 antagonist variants on IL-6-induced STAT signaling in HepG2 cells (pSTATI : left; pSTAT3: right).

[0065] FIGs. 29A-F depict schematic representations of the mono / bivalent single / double armed Fc-based targeted IL-6 antagonist configurations and the untargeted IL-6 antagonist controls.

[0066] FIG. 30 shows the inhibitory capacity of the Fc-based ASGPR VHH-IL-6 antagonist variants (mono / bivalent; single / double armed) on I L-6-induced STAT1 signaling in HepG2 cells.

[0067] FIG. 31 shows the inhibitory capacity of the Fc-based ASGPR VHH-IL-6 antagonist variants (mono / bivalent; single / double armed) on I L-6-induced STAT3 signaling in HepG2 cells.

[0068] FIG. 32 shows the inhibitory capacity of the Fc-based CD8 VHH-IL-6 antagonist variants on I L-6-induced STAT 1 signaling in CD8+ versus CD8- PBMCs.

[0069] FIG. 33 shows the inhibitory capacity of the Fc-based CD8 VHH-IL-6 antagonist variants on I L-6-induced STAT3 signaling in CD8+ versus CD8- PBMCs.

[0070] FIG. 34 shows the inhibitory the Fc-based ASGPR VHH-IL-6 antagonist variants with additional site II mutations on IL-6-induced STAT1 signaling in HepG2 cells.

[0071] FIG. 35 shows the inhibitory capacity of the Fc-based ASGPR VHH-IL-6 antagonist variants with additional site II mutations on I L-6-induced STAT3 signaling in HepG2 cells.

[0072] FIG. 36 shows the inhibitory capacity of Fc-based ASGPR VHH-IL-6 antagonist variants with an additional V121D or S118R_V121D mutation (mono / bivalent; single / double armed as shown in Figs. 29A-F) on IL-6-induced STAT1 signaling in HepG2 cells.

[0073] DB1 / 162518159.2 7 ORN-109PC / 114384-5109

[0074] FIG. 37 shows the Inhibitory capacity of Fc-based ASGPR VHH-Anta6 variants with an additional V121D or S118R_V121D mutation (mono / bivalent; single / double armed as show in Figs. 29A-F) on IL-6-induced STAT3 signaling in HepG2 cells.

[0075] FIG. 38 depicts an overlay of the pSTAT 1 response curves for the different IL-6 antagonist construct configurations (see Figs. 29A-F) with an additional V121 D or S118R_V121 D in the presence of 10 or 100 ng / mL hlL-6.

[0076] FIG. 39 depicts an overlay of the pSTAT3 response curves for the different IL-6 antagonist construct configurations (see Figs. 29A-F) with an additional V121 D or S118R V121D in the presence of 10 or 100 ng / mL hlL-6.

[0077] FIG. 40 shows phosphatase activity was measured using the Phospha-Li ght SEAP Reporter Gene Assay System when HEK-Blue-IL6 or HEK-Blue-IL6-hCLEC9a cells were stimulated with a serial dilution of different IL-6 antagonist-based constructs (in absence or presence of a constant concentration of recombinant human IL-6).

[0078] FIG. 41 depicts STAT3 phosphorylation on primary CLEC9a+ dendritic cells (cDC1) in PBMCs when bivalent, double-armed Fc-based IL-6 antagonist variant was tested for its antagonistic capacity against IL-6.

[0079] FIG. 42 shows pSTATI inhibition in CD8+ T cells when monovalent, single-armed (1x VHH + 1x IL-6 antagonist) and bivalent, double armed (2x VHH + 2x IL-6 antagonist) Fc-based CD8 VHH-IL- 6_Y31G_L57A_S118R_V121D_W157A variants were used.

[0080] DETAILED DESCRIPTION

[0081] In one aspect, the present invention provides a chimeric protein or a chimeric protein complex (including an Fc- based chimeric protein complex) that includes an interleukin-6 (IL-6) antagonist.

[0082] The present invention concerns a target-specific IL-6 antagonist having loss-of-function site II and / or site III mutations that confer reduced or ablated activity and / or affinity for the IL-6 receptor (gp130). In emobodiments, the antagonistic activity of IL-6 still binds strongly when attached to a targeting moiety, said targeting moiety comprising a recognition domain which specifically binds to an antigen or receptor of interest. In embodiments, the target-specific IL-6 antagonist comprises one or more site II and / or site III mutations that inhibit recruitment of two gp 130 receptor chains to the I L-6 / I L-6Ra complex and higher order clustering.

[0083] In various embodiments, the present invention provides a chimeric protein or chimeric protein complex, such as Fc-based chimeric protein complexes, that includes an IL-6 antagonist fused to one or more targeting moieties. In some embodiments, without limitation, the chimeric proteins or chimeric protein complexes target an IL-6 antagonist to hepatocytes, adipose stromal cells, NK cells, CD4 T cells, and CD8 T cells. In vivo, such chimeric proteins or chimeric protein complexes can potently inhibit the response of hepatocytes, adipose stromal cells, NK cells, CD4 T cells, and CDS T cells.

[0084] In various embodiments, the present chimeric proteins and chimeric protein complexes, including Fc-based chimeric protein complexes find use in the treatment of various diseases or disorders such autoimmune diseases or disorders, immune disorders, cancer, hepatocellular carcinoma, pancreatic cancer / melanoma, obesity, Graft- versus-host disease (GVHD), viral infections, cardiovascular diseases, wound, ischemia-related diseases,

[0085] DB1 / 162518159.2 8 ORN-109PC / 114384-5109 neurodegenerative diseases, and / or metabolic diseases and many other diseases and disorders, and the present invention encompasses various methods of treatment.

[0086] In some embodiments, the IL-6 antagonist is a human IL-6 antagonist. In some embodiments, the IL-6 antagonist has ablated or reduced affinity and / or activity for IL-6 receptor (gp130) (e.g., inhibition activity). In some embodiments, the IL-6 antagonist inhibits activity at the IL-6 receptor. In some embodiments, the inhibition activity of the IL-6 antagonist at the IL-6 receptor is induced by attachment to one or more targeting moieties or upon inclusion in the chimeric protein complex.

[0087] In an embodiment, the wild type IL-6 has the amino acid sequence of:

[0088] IL-6 (wild type) (SEQ ID NO: 1)

[0089] MNSFSTSAFGPVAFSLGLLLVLPAAFPAPVPPGEDSKDVAAPHRQPLTSSERIDKQIRYIL DGISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLE FEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQ NQWLQDMTTHLILRSFKEFLQSSLRALRQM.

[0090] In some embodiments, the human IL-6 antagonist has an amino acid sequence of at least 90%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1.

[0091] In some embodiments, the human IL-6 antagonist has the amino acid sequence of wildtype human IL-6.

[0092] Signal peptide cleavage resulting in mature IL-6 has been reported at various positions including cleavage after amino acid 27, 28 or 29. Most commonly accepted in literature is the cleavage after position 28 resulting in the mature IL-6 of SED ID NO: 426:

[0093] PVPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAEN NLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEEQARAVQMSTK VLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKEFLQSSLRAL RQM.

[0094] In some embodiments, the human IL-6 antagonist has an amino acid sequence of at least 90%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 426.

[0095] In some embodiments, the human IL-6 antagonist has one or more substitution mutations at a position selected from Y31 , G35, L57, E59, N60, S118, V121 , and W157, where the residue numbering is based on SEQ ID NO: 426. In some embodiments, one or more of these mutations produce a human IL-6 antagonist with ablated or reduced binding activity and / or affinity for IL-6 receptor (gp130) and / or ablated or reduced biological activity.

[0096] In some embodiments, the mutation comprises a hydrophilic or hydrophobic amino acid residue In embodiments, the hydrophilic amino acid residue is a polar and positively charged hydrophilic residue selected from arginine (R) and lysine (K) or an aromatic, polar and positively charged hydrophilic including histidine (H). In some embodiments, the hydrophilic amino acid residue is a polar and neutral of charge hydrophilic residue selected from

[0097] DB1 / 162518159.2 9 ORN-109PC / 114384-5109 asparagine (N), glutamine (Q), serine (S), threonine (T), proline (P), and cysteine (C). In embodiments, the hydrophilic amino acid residue is a polar and negatively charged hydrophilic residue selected from aspartate (D) and glutamate (E). In some embodiments, the hydrophobic amino acid is a hydrophobic, aliphatic amino acid selected from glycine (G), alanine (A), leucine (L), isoleucine (I), methionine (M), and valine (V) or a hydrophobic, aromatic amino acid selected from phenylalanine (F), tryptophan (W), and tyrosine (Y).

[0098] In some embodiments, the IL-6 antagonist has one or more substitution mutations selected from Y31 D, Y31 F, Y31G, G35F, L57A, L57D, E59F, N60W, S118R, V121D, W157A, and W157G, where the residue numbering is based on SEQ ID NO: 426.

[0099] In embodiments, the IL-6 antagonist has a V121D substitution mutation and further comprises one or more mutations selected from Y31G, L57A, W157A, and W157G, where the residue numbering is based on SEQ ID NO: 426.

[0100] In various embodiments, the mutations allow for IL-6 antagonist to have inhibition activity relative to unmodified or unmutated, i.e., the wild type form of, IL-6 (e.g. comparing IL-6 in a wild type form versus a modified (e.g. mutant) form). Consequentially, in various embodiments, the mutations allow for IL-6 antagonist to have improved safety, e.g. have reduced systemic toxicity, reduced side effects, and reduced off-target effects relative to unmutated, i.e. wild type, IL-6 (e.g. comparing IL-6 in a wild type form versus a modified (e.g. mutant) form).

[0101] In various embodiments, the IL-6 antagonist comprises one or more mutations that reduce or ablate its binding affinity and / or activity for one or more of its receptors. In some embodiments, the IL-6 antagonist comprises one or more mutations that substantially ablate binding affinity or activity for the receptors. In some embodiments, the activity provided by the wild type IL-6 is agonism at the receptor (e.g. activation of a cellular effect at a site of therapy). For example, the wild type IL-6 may activate its receptor. In such embodiments, the mutations of wild type IL-6 result in the IL-6 antagonist having ablated activating activity at the receptor. For example, the mutations may result in the IL-6 antagonist to inhibit an IL-6 activating signal to a target cell.

[0102] In various embodiments, the IL-6 antagonist comprises one or more mutations that cause IL-6 to have ablated affinity, e.g. binding (e.g. KD) and / or activation. In various embodiments, the ablated affinity at I L-6’s receptor allows for inhibition of activity In such embodiments, the IL-6 antagonist has about 1 %, or about 3%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 10%-20%, about 20%-40%, about 50%, about 40%-60%, about 60%-80%, about 80%-100% of the affinity for the receptor relative to the wild type IL-6. In some embodiments, the binding affinity is at least about 2-fold lower, about 3-fold lower, about 4-fold lower, about 5-fold lower, about 6-fold lower, about 7-fold lower, about 8-fold lower, about 9-fold lower, at least about 10-fold lower, at least about 15-fold lower, at least about 20-fold lower, at least about 25-fold lower, at least about 30-fold lower, at least about 35-fold lower, at least about 40-fold lower, at least about 45-fold lower, at least about 50-fold lower, at least about 100-fold lower, at least about 150-fold lower, or about 10-50-fold lower,

[0103] DB1 / 162518159.2 10 ORN-109PC / 114384-5109 about 50-100-fold lower, about 100-150-fold lower, about 150-200-fold lower, or more than 200-fold lower relative to the wild type IL-6.

[0104] In various embodiments, the IL-6 antagonist comprises one or more mutations that reduce the endogenous activity of IL-6 to about 75%, or about 70%, or about 60%, or about 50%, or about 40%, or about 30%, or about 25%, or about 20%, or about 10%, or about 5%, or about 3%, or about 1 %, e.g., relative to the wild type IL-6.

[0105] Receptor binding activity may be measured using methods known in the art For example, affinity and / or binding activity may be assessed by Scatchard plot analysis and computer-fitting of binding data (e.g. Scatchard, 1949) or by reflectometric interference spectroscopy under flow through conditions, as described by Brecht et al. (1993), the entire contents of all of which are hereby incorporated by reference.

[0106] In various embodiments the IL-6 antagonist comprises an amino acid sequence that has at least about 60%, or at least about 61%, or at least about 62%, or at least about 63%, or at least about 64%, or at least about 65%, or at least about 66%, or at least about 67%, or at least about 68%, or at least about 69%, or at least about 70%, or at least about 71%, or at least about 72%, or at least about 73%, or at least about 74%, or at least about 75%, or at least about 76%, or at least about 77%, or at least about 78%, or at least about 79%, or at least about 80%, or at least about 81%, or at least about 82%, or at least about 83%, or at least about 84%, or at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99% sequence identity with the known wild type IL-6 having SEQ ID NO: 1 or mature IL-6 (with a cleaved signal peptide) having SEQ ID NO: 426 (e.g. about 60%, or about 61 %, or about 62%, or about 63%, or about 64%, or about 65%, or about 66%, or about 67%, or about 68%, or about 69%, or about 70%, or about 71%, or about 72%, or about 73%, or about 74%, or about 75%, or about 76%, or about 77%, or about 78%, or about 79%, or about 80%, or about 81 %, or about 82%, or about 83%, or about 84%, or about 85%, or about 86%, or about 87%, or about 88%, or about 89%, or about 90%, or about 91%, or about 92%, or about 93%, or about 94%, or about 95%, or about 96%, or about 97%, or about 98%, or about 99% sequence identity).

[0107] In various embodiments the IL-6 antagonist comprises an amino acid sequence that has at least about 60%, or at least about 61%, or at least about 62%, or at least about 63%, or at least about 64%, or at least about 65%, or at least about 66%, or at least about 67%, or at least about 68%, or at least about 69%, or at least about 70%, or at least about 71%, or at least about 72%, or at least about 73%, or at least about 74%, or at least about 75%, or at least about 76%, or at least about 77%, or at least about 78%, or at least about 79%, or at least about 80%, or at least about 81%, or at least about 82%, or at least about 83%, or at least about 84%, or at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99% sequence identity with any of amino acid sequences of the IL-6 disclosed herein (e.g. about 60%, or about 61%, or about 62%, or about 63%,

[0108] DB1 / 162518159.2 11 ORN-109PC / 114384-5109 or about 64%, or about 65%, or about 66%, or about 67%, or about 68%, or about 69%, or about 70%, or about 71%, or about 72%, or about 73%, or about 74%, or about 75%, or about 76%, or about 77%, or about 78%, or about 79%, or about 80%, or about 81 %, or about 82%, or about 83%, or about 84%, or about 85%, or about 86%, or about 87%, or about 88%, or about 89%, or about 90%, or about 91 %, or about 92%, or about 93%, or about 94%, or about 95%, or about 96%, or about 97%, or about 98%, or about 99% sequence identity)

[0109] In various embodiments, the IL-6 antagonist comprises an amino acid sequence having one or more amino acid mutations. In some embodiments, the one or more amino acid mutations may be independently selected from substitutions, insertions, deletions, and truncations. In some embodiments, the amino acid mutations are amino acid substitutions, and may include conservative and / or non-conservative substitutions.

[0110] “Conservative substitutions” may be made, for instance, on the basis of similarity in polarity, charge, size, solubility, hydrophobicity, hydrophilicity, and / or the amphipathic nature of the amino acid residues involved. The 20 naturally occurring amino acids can be grouped into the following six standard amino acid groups: (1) hydrophobic: Met, Ala, Vai, Leu, lie; (2) neutral hydrophilic: Cys, Ser, Thr; Asn, Gin; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

[0111] As used herein, “conservative substitutions” are defined as exchanges of an amino acid by another amino acid listed within the same group of the six standard amino acid groups shown above. For example, the exchange of Asp by Glu retains one negative charge in the so modified polypeptide. In addition, glycine and proline may be substituted for one another based on their ability to disrupt a-helices.

[0112] As used herein, “non-conservative substitutions” are defined as exchanges of an amino acid by another amino acid listed in a different group of the six standard amino acid groups (1) to (6) shown above.

[0113] In various embodiments, the substitutions may also include non-classical amino acids (e.g. selenocysteine, pyrrolysine, W-formylmethionine [3-alanine, GABA and 6-Aminolevulinic acid, 4-aminobenzoic acid (PABA), D- isomers of the common amino acids, 2,4-diaminobutyric acid, a-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, y-Abu, s-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosme, citrulline, homocitrulline, cysteic acid, t-butylglycine, t- butylalanine, phenylglycine, cyclohexylalanine, p-alanine, fluoro-amino acids, designer amino acids such as |3 methyl amino acids, C a-methyl amino acids, N a-methyl amino acids, and amino acid analogs in general).

[0114] In some embodiments, the inhibition activity of the IL-6 antagonist at the therapeutic receptor is inducible by attachment to a targeting moiety or upon inclusion of a targeting moiety in a chimeric protein or a chimeric protein complex, e g., a Fc-based chimeric protein complex as disclosed herein.

[0115] In embodiments, the chimeric protein or the Fc-based chimeric protein complex of present invention comprises a polypeptide having an amino acid sequence selected from SEQ ID NO: 414-425, 439-456. In embodiments, the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex comprises a His tag

[0116] DB1 / 162518159.2 12 ORN-109PC / 114384-5109 or do not comprise a His tag. In embodiments, the chimeric proteins or chimeric protein complexes such as Fc- based chimeric protein complex comprise a leader sequence or do not comprise a leader sequence

[0117] Targeting Moiety

[0118] In various embodiments, the chimeric protein or chimeric protein complex of the present invention additionally comprise one or more targeting moieties having recognition domains, which specifically bind to a target (e.g. antigen, receptor) of interest. In some embodiments, the chimeric protein or chimeric protein complex may comprise two, three, four, five, six, seven, eight, nine, ten or more targeting moieties. In illustrative embodiments, the chimeric protein or chimeric protein complex of the invention comprise two or more targeting moieties. In such embodiments, the chimeric protein or chimeric protein complex can target two different cells (e.g. to make a synapse) or the same cell (e.g. to get a more concentrated signaling agent effect).

[0119] In various embodiments, the target (e.g. antigen, receptor) of interest can be found on one or more immune cells, which can include, without limitation, T cells, cytotoxic T lymphocytes, T helper cells, natural killer (NK) cells, natural killer T (NKT) cells, anti-tumor or tumor-associated macrophages (e.g. M1 or M2 macrophages), B cells, Breg cells, dendritic cells, adipose stromal cells (ASC), or subsets thereof. In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) of interest and effectively, directly or indirectly, recruit one of more immune cells. In some embodiments, the target (e.g. antigen, receptor) of interest can be found on one or more tumor cells. In some embodiments, the chimeric protein or chimeric protein complex may directly or indirectly recruit an immune cell, e.g., in some embodiments, to a therapeutic site (e.g. a locus with one or more disease cell or cell to be modulated for a therapeutic effect). In some embodiments, the chimeric protein or chimeric protein complex may directly or indirectly recruit an immune cell, e.g. an immune cell that can kill and / or suppress a tumor cell, to a site of action (such as, by way of non-limiting example, the tumor microenvironment).

[0120] In some embodiments, the targeting moiety binds to one or more of the follow targets: CD8, CTLA4, CD3, CD4, DNAM-1 , Nrp1 (neurophilin), TNFR2, GITR, ICOS, CD20, CD70, Clec9a, NKp46, asialoglycoprotein receptor (ASGPR), PD-1 , PD-L1 , PD-L2, SIRPIa, FAP, XCR1 , tenascin, ASC, prostate-specific membrane antigen (PSMA), and ECM proteins.

[0121] In various embodiments, the chimeric protein or chimeric protein complex have targeting moieties having recognition domains, which specifically bind to a target (e.g. antigen, receptor) which is part of a non-cellular structure. In some embodiments, the antigen or receptor is not an integral component of an intact cell or cellular structure. In some embodiments, the antigen or receptor is an extracellular antigen or receptor. In some embodiments, the target is a non-proteinaceous, non-cellular marker, including, without limitation, nucleic acids, inclusive of DNA or RNA, such as, for example, DNA released from necrotic tumor cells or extracellular deposits such as cholesterol.

[0122] In some embodiments, the target (e.g. antigen, receptor) of interest is part of the non-cellular component of the stroma or the extracellular matrix (ECM) or the markers associated therewith As used herein, stroma refers to the connective and supportive framework of a tissue or organ. Stroma may include a compilation of cells such as

[0123] DB1 / 162518159.2 13 ORN-109PC / 114384-5109 fibroblasts / myofibroblasts, glial, epithelia, fat, immune, vascular, smooth muscle, and immune cells along with the extracellular matrix (ECM) and extracellular molecules. In various embodiments, the target (e.g. antigen, receptor) of interest is part of the non-cellular component of the stroma such as the extracellular matrix and extracellular molecules. As used herein, the ECM refers to the non-cellular components present within all tissues and organs. The ECM is composed of a large collection of biochemically distinct components including, without limitation, proteins, glycoproteins, proteoglycans, and polysaccharides. These components of the ECM are usually produced by adjacent cells and secreted into the ECM via exocytosis. Once secreted, the ECM components often aggregate to form a complex network of macromolecules. In various embodiments, the chimeric protein or chimeric protein complex of the invention comprises a targeting moiety that recognizes a target (e.g., an antigen or receptor or non- proteinaceous molecule) located on any component of the ECM. Illustrative components of the ECM include, without limitation, the proteoglycans, the non-proteoglycan polysaccharides, fibers, and other ECM proteins or ECM non-proteins, e.g. polysaccharides and / or lipids, or ECM associated molecules (e.g. proteins or non-proteins, e.g. polysaccharides, nucleic acids and / or lipids).

[0124] In some embodiments, the targeting moiety recognizes a target (e.g. antigen, receptor) on ECM proteoglycans. Proteoglycans are glycosylated proteins. The basic proteoglycan unit includes a core protein with one or more covalently attached glycosaminoglycan (GAG) chains. Proteoglycans have a net negative charge that attracts positively charged sodium ions (Na+), which attracts water molecules via osmosis, keeping the ECM and resident cells hydrated. Proteoglycans may also help to trap and store growth factors within the ECM. Illustrative proteoglycans that may be targeted by the chimeric protein or chimeric protein complex of the invention include, but are not limited to, heparan sulfate, chondroitin sulfate, and keratan sulfate. In an embodiment, the targeting moiety recognizes a target (e.g. antigen, receptor) on non-proteoglycan polysaccharides such as hyaluronic acid.

[0125] In some embodiments, the targeting moiety recognizes a target (e.g. antigen, receptor) on ECM fibers. ECM fibers include collagen fibers and elastin fibers. In some embodiments, the targeting moiety recognizes one or more epitopes on collagens or collagen fibers. Collagens are the most abundant proteins in the ECM. Collagens are present in the ECM as fibrillar proteins and provide structural support to resident cells. In one or more embodiments, the targeting moiety recognizes and binds to various types of collagens present within the ECM including, without limitation, fibrillar collagens (types I, II, III, V, XI), facit collagens (types IX, XII, XIV), short chain collagens (types VIII, X), basement membrane collagens (type IV), and / or collagen types VI, VII, or XIII. Elastin fibers provide elasticity to tissues, allowing them to stretch when needed and then return to their original state. In some embodiments, the target moiety recognizes one or more epitopes on elastins or elastin fibers.

[0126] In some embodiments, the targeting moiety recognizes one or more ECM proteins including, but not limited to, a tenascin, a fibronectin, a fibrin, a laminin, or a nidogen / entactin.

[0127] In an embodiment, the targeting moiety recognizes and binds to tenascin The tenascin (TN) family of glycoproteins includes at least four members, tenascin-C, tenascin-R, tenascin-X, and tenascin W. The primary structures of tenascin proteins include several common motifs ordered in the same consecutive sequence: amino-terminal

[0128] DB1 / 162518159.2 14 ORN-109PC / 114384-5109 heptad repeats, epidermal growth factor (EGF)-like repeats, fibronectin type III domain repeats, and a carboxyl- terminal fibrinogen-like globular domain. Each protein member is associated with typical variations in the number and nature of EGF-like and fibronectin type III repeats. Isoform variants also exist particularly with respect to tenascin-C Over 27 splice variants and / or isoforms of tenascin-C are known. In a particular embodiment, the targeting moiety recognizes and binds to tenascin-CA1 . Similarly, tenascin-R also has various splice variants and isoforms Tenascin-R usually exists as dimers or trimers. Tenascin-X is the largest member of the tenascin family and is known to exist as trimers. Tenascin-W exists as trimers. In some embodiments, the targeting moiety recognizes one or more epitopes on a tenascin protein. In some embodiments, the targeting moiety recognizes the monomeric and / or the dimeric and / or the trimeric and / or the hexameric forms of a tenascin protein.

[0129] In an embodiment, the targeting moieties recognize and bind to fibronectin. Fibronectins are glycoproteins that connect cells with collagen fibers in the ECM, allowing cells to move through the ECM. Upon binding to integrins, fibronectins unfolds to form functional dimers In some embodiments, the targeting moiety recognizes the monomeric and / or the dimeric forms of fibronectin. In some embodiments, the targeting moiety recognizes one or more epitopes on fibronectin. In illustrative embodiments, the targeting moiety recognizes fibronectin extracellular domain A (EDA) or fibronectin extracellular domain B (EDB). Elevated levels of EDA are associated with various diseases and disorders including psoriasis, rheumatoid arthritis, diabetes, and cancer. In some embodiments, the targeting moiety recognizes fibronectin that contains the EDA isoform and may be utilized to target the chimeric protein or chimeric protein complex to diseased cells including cancer cells. In some embodiments, the targeting moiety recognizes fibronectin that contains the EDB isoform. In various embodiments, such targeting moieties may be utilized to target the chimeric protein or chimeric protein complex to tumor cells including the tumor neovasculature.

[0130] In an embodiment, the targeting moiety recognizes and binds to fibrin. Fibrin is another protein substance often found in the matrix network of the ECM. Fibrin is formed by the action of the protease thrombin on fibrinogen, which causes the fibrin to polymerize. In some embodiments, the targeting moiety recognizes one or more epitopes on fibrin. In some embodiments, the targeting moiety recognizes the monomeric as well as the polymerized forms of fibrin.

[0131] In an embodiment, the targeting moiety recognizes and binds to laminin Laminin is a major component of the basal lamina, which is a protein network foundation for cells and organs Laminins are heterotrimeric proteins that contain an a-chain, a [3-chain, and a y-chain. In some embodiments, the targeting moiety recognizes one or more epitopes on laminin. In some embodiments, the targeting moiety recognizes the monomeric, the dimeric as well as the trimeric forms of laminin.

[0132] In an embodiment, the targeting moiety recognizes and binds to a nidogen or entactin. Nidogens / entactins are a family of highly conserved, sulfated glycoproteins. They make up the major structural component of the basement membranes and function to link laminin and collagen IV networks in basement membranes. Members of this family

[0133] DB1 / 162518159.2 15 ORN-109PC / 114384-5109 include nidogen-1 and nidogen-2. In various embodiments, the targeting moiety recognizes an epitope on nidogen- 1 and / or nidogen-2.

[0134] In various embodiments, the targeting moiety comprises an antigen recognition domain that recognizes an epitope present on any of the targets described herein In an embodiment, the antigen-recognition domain recognizes one or more linear epitopes present on the protein. As used herein, a linear epitope refers to any continuous sequence of amino acids present on the protein. In another embodiment, the antigen-recognition domain recognizes one or more conformational epitopes present on the protein. As used herein, a conformation epitope refers to one or more sections of amino acids (which may be discontinuous) which form a three-dimensional surface with features and / or shapes and / or tertiary structures capable of being recognized by an antigen recognition domain.

[0135] In various embodiments, the targeting moiety may bind to the full-length and / or mature forms and / or isoforms and / or splice variants and / or fragments and / or any other naturally occurring or synthetic analogs, variants, or mutants of any of the targets described herein. In various embodiments, the targeting moiety may bind to any forms of the proteins described herein, including monomeric, dimeric, trimeric, tetrameric, heterodimeric, multimeric and associated forms. In various embodiments, the targeting moiety may bind to any post-transl ationally modified forms of the proteins described herein, such as glycosylated and / or phosphorylated forms.

[0136] In various embodiments, the targeting moiety comprises an antigen recognition domain that recognizes extracellular molecules such as DNA. In some embodiments, the targeting moiety comprises an antigen recognition domain that recognizes DNA. In an embodiment, the DNA is shed into the extracellular space from necrotic or apoptotic tumor cells or other diseased cells.

[0137] In various embodiments, the targeting moiety comprises an antigen recognition domain that recognizes one or more non-cellular structures associated with atherosclerotic plaques. Two types of atherosclerotic plaques are known. The fibro-lipid (fibro-fatty) plaque is characterized by an accumulation of lipid-laden cells underneath the intima of the arteries Beneath the endothelium there is a fibrous cap covering the atheromatous core of the plaque. The core includes lipid-laden cells (macrophages and smooth muscle cells) with elevated tissue cholesterol and cholesterol ester content, fibrin, proteoglycans, collagen, elastin, and cellular debris In advanced plaques, the central core of the plaque usually contains extracellular cholesterol deposits (released from dead cells), which form areas of cholesterol crystals with empty, needle-like clefts At the periphery of the plaque are younger foamy cells and capillaries. A fibrous plaque is also localized under the intima, within the wall of the artery resulting in thickening and expansion of the wall and, sometimes, spotty localized narrowing of the lumen with some atrophy of the muscular layer. The fibrous plaque contains collagen fibers (eosinophilic), precipitates of calcium (hematoxylinophilic) and lipid-laden cells. In some embodiments, the targeting moiety recognizes and binds to one or more of the non-cellular components of these plaques such as the fibrin, proteoglycans, collagen, elastin, cellular debris, and calcium or other mineral deposits or precipitates. In some embodiments, the cellular debris is a nucleic acid, e.g. DNA or RNA, released from dead cells.

[0138] DB1 / 162518159.2 16 ORN-109PC / 114384-5109

[0139] In various embodiments, the targeting moiety comprises an antigen recognition domain that recognizes one or more non-cellular structures found in the brain plaques associated with neurodegenerative diseases. In some embodiments, the targeting moiety recognizes and binds to one or more non-cellular structures located in the amyloid plaques found in the brains of patients with Alzheimer’s disease. For example, the targeting moiety may recognize and bind to the peptide amyloid beta, which is a major component of the amyloid plaques. In some embodiments, the targeting moiety recognizes and binds to one or more non-cellular structures located in the brains plaques found in patients with Huntington’s disease. In various embodiments, the targeting moiety recognizes and binds to one or more non-cellular structures found in plaques associated with other neurodegenerative or musculoskeletal diseases such as Lewy body dementia and inclusion body myositis.

[0140] In some embodiments, the chimeric protein or chimeric protein complex of the invention may have two or more targeting moieties that bind to non-cellular structures. In some embodiments, there are two targeting moieties and one targets a cell while the other targets a non-cellular structure In various embodiments, the targeting moieties can directly or indirectly recruit cells, such as disease cells and / or effector cells. In some embodiments, chimeric protein or chimeric protein complex are capable of, or find use in, methods involving, shifting the balance of immune cells in favor of immune attack of a tumor For instance, chimeric protein or chimeric protein complex can shift the ratio of immune cells at a site of clinical importance in favor of cells that can kill and / or suppress a tumor (e.g. T cells, cytotoxic T lymphocytes, T helper cells, natural killer (NK) cells, natural killer T (NKT) cells, anti-tumor macrophages (e.g. M1 macrophages), B cells, dendritic cells, or subsets thereof) and in opposition to cells that protect tumors (e.g. myeloid-derived suppressor cells (MDSCs), regulatory T cells (Tregs); tumor associated neutrophils (TANs), M2 macrophages, tumor associated macrophages (TAMs), or subsets thereof). In some embodiments, chimeric protein or chimeric protein complex are capable of increasing a ratio of effector T cells to regulatory T cells.

[0141] For example, in some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with T cells. In some embodiments, the recognition domains directly or indirectly recruit T cells. In an embodiment, the recognition domains specifically bind to effector T cells. In some embodiments, the recognition domain directly or indirectly recruits effector T cells, e.g., in some embodiments, to a therapeutic site (e.g. a locus with one or more disease cell or cell to be modulated for a therapeutic effect). Illustrative effector T cells include cytotoxic T cells (e.g. o|3 TCR, CD3-, CD8-, CD45RO ); CD4- effector T cells (e.g. o|3 TCR, CD3\ CD4-, CCR7-, CD62Lhi, IL7R / CD127-); CD8+effector T cells (e.g. a(3 TCR, CD3+, CD8+, CCR7+, CD62Lhi, IL7R / CD127+); effector memory T cells (e.g. CD62Llow, CD44-, TCR, CD3+, IL7R / CD127-, IL-15R-, CCR7low); central memory T cells (e.g. CCR7+, CD62L+, CD27+; or CCR7hi, CD44+, CD62Lhi, TCR, CD3+, IL-7R / CD127+, IL-15R+); CD62L+effector T cells; CD8+effector memory T cells (TEM) including early effector memory T cells (CD27- CD62L-) and late effector memory T cells (CD27- CD62L-) (TemE and TemL, respectively); CD127(+)CD25(low / -) effector T cells; CD127(jCD25(j effector T cells; CD8+stem cell memory effector cells (TSCM) (e.g. CD44(low)CD62L(high)CD122(high)sca(+)); TH1 effector T-cells (e.g. CXCR3+, CXCR6+and CCR5+; or a|3 TCR, CD3+, CD4+, I L-12R+, IFNyR-, CXCR3+), TH2 effector T cells (e.g. CCR3+, CCR4+and CCR8+; or a0 TCR, CD3+,

[0142] DB1 / 162518159.2 17 ORN-109PC / 114384-5109

[0143] CD4-, I L-4R+, IL-33R-, CCR4+, I L-17RB+, CRTH2+); TH9 effector T cells (e.g. a0 TCR, CD3+, CD4+); TH17 effector T cells (e.g. a0 TCR, CD3+, CD4+, IL-23R+, CCR6+, IL-1R+); CD4+CD45RO+CCR7+effector T cells, ICOS+effector T cells; CD4*CD45RO*CCR7( ) effector T cells; and effector T cells secreting IL-2, IL-4 and / or IFN-y.

[0144] Illustrative T cell antigens of interest include, for example (and inclusive of the extracellular domains, where applicable): CD8, CD3, SLAMF4, IL-2Ra, 4-1 BB / TN FRSF9, IL-2 R 0, ALCAM, B7-1, IL-4 R, B7-H3, BLAME / SLAMFS, CEACAM1 , IL-6 R, CCR3, IL-7 Ra, CCR4, CXCRI / IL-S RA, CCR5, CCR6, IL-10R a, CCR 7, IL- I 0 R [3, CCRS, IL-12 R 0 1, CCR9, IL-12 R 0 2, CD2, IL-13 R a 1, IL-13, CD3, CD4, ILT2 / CDS5j, ILT3 / CDS5k, ILT4 / CDS5d, ILT5 / CDS5a, lutegrin a 4 / CD49d, CDS, Integrin a E / CD103, CD6, Integrin a M / CD 11 b, CDS, Integrin a X / CD11c, Integrin 0 2 / CDIS, KIR / CD15S, CD27 / TNFRSF7, KIR2DL1, CD2S, KIR2DL3, CD30 / TNFRSFS, KIR2DL4 / CD15Sd, CD31 / PECAM-1, KIR2DS4, CD40 Ligand / TNFSF5, LAG-3, CD43, LAIR1 , CD45, LAIR2, CDS3, Leukotriene B4-R1 , CDS4 / SLAMF5, NCAM-L1 , CD94, NKG2A, CD97, NKG2C, CD229 / SLAMF3, NKG2D, CD2F-10 / SLAMF9, NT-4, CD69, NTB-A / SLAMF6, Common y Chain / IL-2 R y, Osteopontin, CRACC / SLAMF7, PD-1, CRTAM, PSGL-1, CTLA-4, RANK / TNFRSF11A, CX3CR1 , CX3CL1, L- Selectin, CXCR3, SIRP 0 1, CXCR4, SLAM, CXCR6, TCCR / WSX-1, DNAM-1, Thymopoietin, EMMPRIN / CD147, TIM-1 , EphB6, TIM-2, Fas / TNFRSF6, TIM-3, Fas Ligand / TNFSF6, TIM-4, Fey RIII / CD16, TIM-6, TNFR1 / TNFRSF1A, Granulysin, TNF RIII / TNFRSF1B, TRAIL RI / TNFRSFIOA, ICAM-1 / CD54, TRAIL R2 / TNFRSF10B, ICAM-2 / CD102, TRAILR3 / TNFRSF10C,IFN-yR1 , TRAILR4 / TNFRSF10D, IFN-y R2, TSLP, IL-1 R1 and TSLP R. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these illustrative T cell antigens

[0145] By way of non-limiting example, in various embodiments, the chimeric protein or chimeric protein complex have a targeting moiety directed against a checkpoint marker expressed on a T cell, e.g. one or more of PD-1 , CD28, CTLA4, ICOS, BTLA, KIR, LAG3, CD137, 0X40, CD27, CD40L, TIM3, and A2aR.

[0146] For example, in some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with B cells In some embodiments, the recognition domains directly or indirectly recruit B cells, e.g., in some embodiments, to a therapeutic site (e.g. a locus with one or more disease cell or cell to be modulated for a therapeutic effect). Illustrative B cell antigens of interest include, for example, CD10, CD19, CD20, CD21, CD22, CD23, CD24, CD37, CD38, CD39, CD40, CD70, CD72, CD73, CD74, CDw75, CDw76, CD77, CD78, CD79a / b, CD80, CD81 , CD82, CD83, CD84, CD85, CD86, CD89, CD98, CD126, CD127, CDw130, CD138, CDw150, CS1 , and B-cell maturation antigen (BCMA). In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these illustrative B cell antigens

[0147] By way of further example, in some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with Natural Killer cells. In some embodiments, the recognition domains directly or indirectly recruit Natural Killer cells, e.g. , in some embodiments, to a therapeutic site (e.g. a locus with one or more disease cell or cell to be modulated for a therapeutic effect). Illustrative Natural Killer cell antigens of interest include, for example TIGIT, 2B4 / SLAMF4, KIR2DS4, CD155 / PVR, KIR3DL1, CD94, LMIR1 / CD300A, CD69,

[0148] DB1 / 162518159.2 18 ORN-109PC / 114384-5109

[0149] LMIR2 / CD300c, CRACC / SLAMF7, LM I R3 / CD300LF, DNAM-1, LMIR5 / CD300LB, Fc-epsilon RII, LMIR6 / CD300LE, Fc-y RI / CD64, MICA, Fc-y RIIB / CD32b, MICB, Fc-y RIIC / CD32c, MULT-1 , Fc-y RIIA / CD32a, Nectin-2 / CD112, Fc-y RIII / CD16, NKG2A, FcRH 1 / IRTA5, NKG2C, FcRH2 / IRTA4, NKG2D, FcRH4 / IRTA1, NKp30, FcRH5 / IRTA2, NKp44, Fc-Receptor-like 3 / CD16-2, NKp46 / NCR1, NKp80 / KLRF1, NTB-A / SLAMF6, Rae-1 , Rae-1 a, Rae-1 p, Rae-1 delta, H60, Rae-1 epsilon, ILT2 / CD85j, Rae-1 y, ILT3 / CD85k, TREM-1, ILT4 / CD85d, TREM-2, ILT5 / CD85a, TREM-3, KIR / CD158, TREML1 / TLT-1 , KIR2DL1 , ULBP-1, KIR2DL3, ULBP-2, KIR2DL4 / CD 158d and ULBP-3. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these illustrative NK cell antigens.

[0150] Also, in some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with macrophages / monocytes. In some embodiments, the recognition domains directly or indirectly recruit macrophages / monocytes, e.g., in some embodiments, to a therapeutic site (e.g. a locus with one or more disease cell or cell to be modulated for a therapeutic effect). Illustrative macrophages / monocyte antigens of interest include, for example SIRPI a, B7-1 / CD80, ILT4 / CD85d, B7-H1 , ILT5 / CD85a, Common p Chain, Integrin a 4 / CD49d, BLAME / SLAMF8, Integrin a X / CDIIc, CCL6 / C10, Integrin p 2 / CD18, CD155 / PVR, Integrin p 3 / CD61 , CD31 / PECAM-1 , Latexin, CD36 / SR-B3, Leukotriene B4 R1, CD40 / TNFRSF5, LIMPIIISR-B2, CD43, LMIR1 / CD300A, CD45, LMIR2 / CD300c, CD68, LMIR3 / CD300LF, CD84 / SLAMF5, LMIR5 / CD300LB, CD97, LMIR6 / CD300LE, CD163, LRP-1 , CD2F-10 / SLAMF9, MARCO, CRACC / SLAMF7, MD-1, ECF-L, MD-2, EMMPRIN / CD147, MGL2, Endoglin / CD105, Osteoactivin / GPNMB, Fc-y RI / CD64, Osteopontin, Fc-y RIIB / CD32b, 4 / CD49d, CCR5, Integrin a M / CDII b, CCR8, Integrin a X / CDIIc, CD155 / PVR, Integrin p 2 / CD18, CD14, Integrin p 3 / CD61, CD36 / SR-B3, LAIR1, CD43, LAIR2, CD45, Leukotriene B4-R1, CD68, LIMPIIISR-B2, CD84 / SLAMF5, LM IR1 / CD300A, CD97, LMIR2 / CD300c, LMIR3 / CD300LF, Coagulation Factor II l / Tissue Factor, LMIR5 / CD300LB, CX3CR1 , CX3CL1 , LMIR6 / CD300LE, CXCR4, LRP-1 , CXCR6, M-CSF R, DEP-1 / CD148, MD-1 , DNAM-1 , MD-2, EMMPRIN / CD147, MMR, Endoglin / CD105, NCAM-L1 , Fc-y RI / CD64, PSGL-1, Fc-y RIIIICD16, RP105, G-CSF R, L-Selectin, GM-CSF R a, Siglec-3 / CD33, HVEM / TNFRSF14, SLAM, ICAM-1 / CD54, TCCR / WSX-1 , ICAM- 2 / CD102, TREM-I, IL-6 R, TREM-2, CXCRI / IL-8 RA, TREM-3 and TREMLI / TLT-1 . In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these illustrative macrophage / monocyte antigens.

[0151] Also, in some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with dendritic cells. In some embodiments, the recognition domains directly or indirectly recruit dendritic cells, e.g., in some embodiments, to a therapeutic site (e.g. a locus with one or more disease cell or cell to be modulated for a therapeutic effect). Illustrative dendritic cell antigens of interest include, for example, CLEC9A, XCR1 , RANK, CD36 / SRB3, LOX-1 / SR-E1, CD68, MARCO, CD163, SR-A1 / MSR, CD5L, SREC-1, CL-PI / COLEC12, SREC-II,

[0152] DB1 / 162518159.2 19 ORN-109PC / 114384-5109

[0153] LIMPIIISRB2, RP105, TLR4, TLR1 , TLR5, TLR2, TLR6, TLR3, TLR9, 4-IBB Ligand / TN FSF9, IL-12 / IL-23 p40, 4- Amino-1,8-naphthalimide, ILT2 / CD85j, CCL21 / 6Ckine, ILT3 / CD85k, 8-oxo-dG, ILT4 / CD85d, 8D6A, ILT5 / CD85a, A2B5, lutegrin a 4 / CD49d, Aag, Integrin p 2 / CD18, AMIGA, Langerin, B7-2 / CD86, Leukotriene B4 Rl, B7-H3, LMIR1 / CD300A, BLAME / SLAMF8, LMIR2 / CD300c, Clq R1 / CD93, LMIR3 / CD300LF, CCR6, LMIR5 / CD300LB CCR7, LMIR6 / CD300LE, CD40 / TNFRSF5, MAG / Siglec-4-a, CD43, MCAM, CD45, MD-1, CD68, MD-2, CD83, MDL-1 / CLEC5A, CD84 / SLAMF5, MMR, CD97, NCAMLI, CD2F-10 / SLAMF9, Osteoactivin GPNMB, Chern 23, PD- L2, CLEC-1, RP105, CLEC-2, CLEC-8, Siglec-2 / CD22, CRACC / SLAMF7, Siglec-3 / CD33, DC-SIGN, Siglec-5, DC- SIGNR / CD299, Siglec-6, DCAR, Siglec-7, DCIR / CLEC4A, Siglec-9, DEC-205, Siglec-10, Dectin-1 / CLEC7A, Siglec-F, Dectin-2 / CLEC6A, SIGNR1 / CD209, DEP-1 / CD148, SIGNR4, DLEC / CLEC4C, SLAM, EMMPRIN / CD147, TCCR / WSX-1, Fc-y R1 / CD64, TLR3, Fc-y RIIB / CD32b, TREM-1 , Fc-y RIIC / CD32c, TREM-2, Fc-y RIIA / CD32a, TREM-3, Fc-y RIII / CD16, TREML1 / TLT-1, ICAM-2 / CD102 and Vanilloid R1. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these illustrative DC antigens.

[0154] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) on immune cells selected from, but not limited to, megakaryocytes, thrombocytes, erythrocytes, mast cells, basophils, neutrophils, myeloid cells, monocytes, eosinophils, or subsets thereof. In some embodiments, the recognition domains directly or indirectly recruit megakaryocytes, thrombocytes, erythrocytes, mast cells, basophils, neutrophils, myeloid cells, monocytes, eosinophils, or subsets thereof, e.g., in some embodiments, to a therapeutic site (e.g. a locus with one or more disease cell or cell to be modulated for a therapeutic effect) In some embodiments, the immune cell is selected from a T cell, a B cell, a dendritic cell, a macrophage, a neutrophil, a mast cell, a monocyte, a red blood cell, myeloid cell, myeloid derived suppressor cell, a NKT cell, and a NK cell, or derivatives thereof.

[0155] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with megakaryocytes and / or thrombocytes. Illustrative megakaryocyte and / or thrombocyte antigens of interest include, for example, GP llb / llla, GPIb, vWF, PF4, and TSP. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these illustrative megakaryocyte and / or thrombocyte antigens

[0156] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with erythrocytes. Illustrative erythrocyte antigens of interest include, for example, CD34, CD36, CD38, CD41 a (platelet glycoprotein llb / llla), CD41 b (GPIIb), CD71 (transferrin receptor), CD105, glycophorin A, glycophorin C, c-kit, HLA- DR, H2 (MHC-II), and Rhesus antigens. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these illustrative erythrocyte antigens.

[0157] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with mast cells. Illustrative mast cells antigens of interest include, for example, SCFR / CD117, FceRI, CD2, CD25, CD35,

[0158] DB1 / 162518159.2 20 ORN-109PC / 114384-5109

[0159] CD88, CD203c, C5R1, CMAI, FCERIA, FCER2, TPSABI. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these mast cell antigens.

[0160] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with basophils. Illustrative basophils antigens of interest include, for example, FcERI, CD203c, CD123, CD13, CD107a, CD107b, and CD164. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these basophil antigens.

[0161] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with neutrophils. Illustrative neutrophils antigens of interest include, for example, 7D5, CD10 / CALLA, CD13, CD16 (FcRIII), CD18 proteins (LFA-1, CR3, and p150, 95), CD45, CD67, and CD177. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these neutrophil antigens.

[0162] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with eosinophils. Illustrative eosinophils antigens of interest include, for example, CD35, CD44 and CD69 In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these eosinophil antigens.

[0163] In various embodiments, the recognition domain may bind to any appropriate target, antigen, receptor, or cell surface markers known by the skilled artisan. In some embodiments, the antigen or cell surface marker is a tissuespecific marker. Illustrative tissue-specific markers include, but are not limited to, endothelial cell surface markers such as ACE, CD14, CD34, CDH5, ENG, ICAM2, MCAM, NOS3, PECAMI, PROCR, SELE, SELP, TEK, THBD, VCAMI, VWF; smooth muscle cell surface markers such as ACTA2, MYHIO, MYH1 1 , MYH9, MYOCD; fibroblast (stromal) cell surface markers such as ALCAM, CD34, COLIAI, COL1A2, COL3A1, FAP, PH-4; epithelial cell surface markers such as CDID, K6IRS2, KRTIO, KRT13, KRT17, KRT18, KRT19, KRT4, KRT5, KRT8, MUCI, TACSTDI; neovasculature markers such as CD13, TFNA, Alpha-v beta-3 (dv ), E-selectin; and adipocyte surface markers such as ADIPOQ, FABP4, and RETN. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these antigens. In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of cells having these antigens

[0164] In some embodiments, the recognition domains specifically bind to a target (e.g. antigen, receptor) associated with tumor cells. In some embodiments, the recognition domains directly or indirectly recruit tumor cells. For instance, in some embodiments, the direct or indirect recruitment of the tumor cell is to one or more effector cell (e.g. an immune cell as described herein) that can kill and / or suppress the tumor cell

[0165] Tumor cells or cancer cells refer to an uncontrolled growth of cells or tissues and / or an abnormal increase in cell survival and / or inhibition of apoptosis, which interferes with the normal functioning of bodily organs and systems. For example, tumor cells include benign and malignant cancers, polyps, hyperplasia, as well as dormant tumors or micrometastases. Illustrative tumor cells include, but are not limited to cells of: basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and central nervous system cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the

[0166] DB1 / 162518159.2 21 ORN-109PC / 114384-5109 digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma; hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or renal cancer; larynx cancer; leukemia; liver cancer; lung cancer (e.g., small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); melanoma; myeloma; neuroblastoma; oral cavity cancer (lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary gland carcinoma; sarcoma; skin cancer; squamous cell cancer; stomach cancer; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary system; vulval cancer; lymphoma including Hodgkin's and non-Hodgkin's lymphoma, as well as B-cell lymphoma (including low grade / follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade / follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblastic leukemia; as well as other carcinomas and sarcomas; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal vascular proliferation associated with phakomatoses, edema (e.g. that associated with brain tumors), and Meigs' syndrome.

[0167] Tumor cells, or cancer cells also include, but are not limited to, carcinomas, e.g. various subtypes, including, for example, adenocarcinoma, basal cell carcinoma, squamous cell carcinoma, and transitional cell carcinoma), sarcomas (including, for example, bone and soft tissue), leukemias (including, for example, acute myeloid, acute lymphoblastic, chronic myeloid, chronic lymphocytic, and hairy cell), lymphomas and myelomas (including, for example, Hodgkin and non-Hodgkin lymphomas, light chain, non-secretory, MGUS, and plasmacytomas), and central nervous system cancers (including, for example, brain (e.g. gliomas (e.g. astrocytoma, oligodendroglioma, and ependymoma), meningioma, pituitary adenoma, and neuromas, and spinal cord tumors (e.g. meningiomas and neurofibroma).

[0168] Illustrative tumor antigens include, but are not limited to, MART-1 / Melan-A, gp100, Dipeptidyl peptidase IV (DPPIV), adenosine deaminase-binding protein (ADAbp), cyclophilin b, Colorectal associated antigen (CRC)-0017- 1A / GA733, Carcinoembryonic Antigen (CEA) and its immunogenic epitopes CAP-1 and CAP-2, etv6, aml1 , Prostate Specific Antigen (PSA) and its immunogenic epitopes PSA-1, PSA-2, and PSA-3, prostate-specific membrane antigen (PSMA), T-cell receptor / CD3-zeta chain, MAGE-family of tumor antigens (e.g., MAGE-A1 , MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE- A11, MAGE-A12, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE-C1 , MAGE- 02, MAGE-C3, MAGE-C4, MAGE-C5), GAGE-family of tumor antigens (e.g., GAGE-1 , GAGE-2, GAGE-3, GAGE- 4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, GAGE-9), BAGE, RAGE, LAGE-1 , NAG, GnT-V, MUM-1, CDK4, tyrosinase, p53, MUC family, HER2 / neu, p21 ras, RCAS1, a-fetoprotein, E-cadherin, a-catenin, [3-catenin and y- catenin, p120ctn, gp100 Pmell 17, PRAME, NY-ESO-1, cdc27, adenomatous polyposis coli protein (APC), fodrin, Connexin 37, Ig-idiotype, p15, gp75, GM2 and GD2 gangliosides, viral products such as human papilloma virus

[0169] DB1 / 162518159.2 22 ORN-109PC / 114384-5109 proteins, Smad family of tumor antigens, Imp-1, NA, EBV-encoded nuclear antigen (EBNA)-1 , brain glycogen phosphorylase, SSX-1, SSX-2 (HOM-MEL-40), SSX-1, SSX-4, SSX-5, SCP-1 CT-7, c-erbB-2, CD19, CD20, CD22, CD30, CD33, CD37, CD47, CS1, CD38, ASGPR, CD56, CD70, CD74, CD138, AGS16, MUC1 , GPNMB, Ep-CAM, PD-L1, PD-L2, PMSA, and BCMA (TNFRSF17). In various embodiments, a targeting moiety of the chimeric protein or chimeric protein complex binds one or more of these tumor antigens. In an embodiment, the chimeric protein or chimeric protein complex binds to HER2. In another embodiment, the chimeric protein or chimeric protein complex binds to PD-L2.

[0170] In some embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have (i) one or more of the targeting moieties which is directed against an immune cell selected from a T cell, a B cell, a dendritic cell, a macrophage, a NK cell, or subsets thereof and (ii) one or more of the targeting moieties which is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In one embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have (i) a targeting moiety directed against a T cell (including, without limitation an effector T cell) and (ii) a targeting moiety is directed against a tumor cell, along with any of the signaling agents described herein. In one embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have (i) a targeting moiety directed against a B cell and (ii) a targeting moiety is directed against a tumor cell, along with any of the signaling agents described herein. In one embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have (i) a targeting moiety directed against a dendritic cell and (ii) a targeting moiety is directed against a tumor cell, along with any of the signaling agents described herein. In one embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have (i) a targeting moiety directed against a macrophage and (ii) a targeting moiety is directed against a tumor cell, along with any of the signaling agents described herein. In one embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have (i) a targeting moiety directed against a NK cell and (ii) a targeting moiety is directed against a tumor cell, along with any of the signaling agents described herein.

[0171] By way of non-limiting example, in various embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have (i) a targeting moiety directed against a T cell, for example, mediated by targeting to CD8, SLAMF4, IL-2 R a, 4-1 BB / TNFRSF9, IL-2 R 0, ALCAM, B7-1, IL-4 R, B7- H3, BLAME / SLAMFS, CEACAM1, IL-6 R, CCR3, IL-7 Ra, CCR4, CXCRI / IL-S RA, CCR5, CCR6, IL-10R a, CCR 7, IL-1 0 R 0, CCRS, IL-12 R 01 , CCR9, IL-12 R 02, CD2, IL-13 R a 1 , IL-13, CD3, CD4, ILT2 / CDS5j, ILT3 / CDS5k, ILT4 / CDS5d, ILT5 / CDS5a, lutegrin a 4 / CD49d, CDS, Integrin a E / CD103, CD6, Integrin a M / CD 11 b, CDS, Integrin a X / CD11c, Integrin 0 2 / CDIS, KIR / CD15S, CD27 / TNFRSF7, KIR2DL1, CD2S, KIR2DL3, CD30 / TNFRSFS, KIR2DL4 / CD15Sd, CD31 / PECAM-1, KIR2DS4, CD40 Ligand / TNFSF5, LAG-3, CD43, LAIR1 , CD45, LAIR2, CDS3, Leukotriene B4-R1 , CDS4 / SLAMF5, NCAM-L1 , CD94, NKG2A, CD97, NKG2C, CD229 / SLAMF3, NKG2D, CD2F-10 / SLAMF9, NT-4, CD69, NTB-A / SLAMF6, Common y Chain / IL-2 R y, Osteopontin, CRACC / SLAMF7, PD-1, CRTAM, PSGL-1, CTLA-4, RANK / TNFRSF11A, CX3CR1, CX3CL1 , L-

[0172] DB1 / 162518159.2 23 ORN-109PC / 114384-5109

[0173] Selectin, CXCR3, SIRP 1, CXCR4, SLAM, CXCR6, TCCR / WSX-1, DNAM-1, Thymopoietin, EMMPRIN / CD147, TIM-1 , EphB6, TIM-2, Fas / TNFRSF6, TIM-3, Fas Ligand / TNFSF6, TIM-4, Fey RIII / CD16, TIM-6, TNFR1 / TNFRSF1A, Granulysin, TNF RIII / TNFRSF1B, TRAIL RI / TNFRSFIOA, ICAM-1 / CD54, TRAIL R2 / TNFRSF10B, ICAM-2 / CD102, TRAILR3 / TNFRSF10C,IFN-yR1, TRAILR4 / TNFRSF10D, IFN-y R2, TSLP, IL-1 R1, or TSLP R; and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0174] By way of non-limiting example, in various embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have a targeting moiety directed against (i) a checkpoint marker expressed on a T cell, e.g. one or more of PD-1, CD28, CTLA4, IGOS, BTLA, KIR, LAG3, CD137, 0X40, CD27, CD40L, TIM3, and A2aR and (ii) a targeting moiety is directed against a tumor cell, along with any of the signaling agents described herein.

[0175] In various embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have one or more targeting moieties directed against PD-1. In some embodiments, the chimeric protein or chimeric protein complex have one or more targeting moieties, which selectively bind a PD-1 polypeptide. In some embodiments, the chimeric protein or chimeric protein complex comprise one or more antibodies, antibody derivatives or formats, peptides or polypeptides, or fusion proteins that selectively bind a PD-1 polypeptide.

[0176] In an embodiment, the targeting moiety comprises the anti-PD-1 antibody, nivolumab (aka BMS-936558, MDX- 1106, ONO-4538, OPDIVO), or fragments thereof. Nivolumab (clone 504) and other human monoclonal antibodies that specifically bind to PD-1 are disclosed in US 8,008,449 and WO 2006 / 121168, the entire disclosures of which are hereby incorporated by reference In illustrative embodiments, nivolumab or an antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEO ID NO: 9 and / or a light chain comprising the amino acid sequence of SEO ID NO: 10.

[0177] In an embodiment, the targeting moiety comprises the anti-PD-1 antibody pidilizumab (aka CT-011 , hBAT or hBAT- 1), or fragments thereof. Pidilizumab and other humanized anti-PD-1 monoclonal antibodies are disclosed in US 2008 / 0025980 and WO 2009 / 101611 , the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a light chain variable regions comprising an amino acid sequence selected from SEO ID NOS: 15-18 of US 2008 / 0025980: SEO ID No: 15 of US 2008 / 0025980 (SEO ID NO: 11); SEQ ID No: 16 of US 2008 / 0025980(SEQ ID NO: 12);SEQ ID No: 17 of US 2008 / 0025980 (SEQ ID NO: 13); and SEQ ID No: 18 of US 2008 / 0025980 (SEQ ID NO: 14) ;and / or a heavy chain comprising an amino acid sequence selected from SEQ ID NOS: 20-24 of US 2008 / 0025980: SEQ ID No: 20 of US 2008 / 0025980 (SEQ ID NO: 15); SEQ ID No: 21 of US 2008 / 0025980 (SEQ ID NO: 16); SEQ ID No: 22 of US 2008 / 0025980 (SEQ ID NO: 17); SEQ ID No: 23 of US 2008 / 0025980 (SEQ ID NO: 18); and SEQ ID No: 24 of US 2008 / 0025980 (SEQ ID NO: 19).

[0178] In an embodiment, the targeting moiety comprises a light chain comprising SEQ ID NO: 18 of US 2008 / 0025980 (SEQ ID NO: 14) and a heavy chain comprising SEQ ID NO: 22 of US 2008 / 0025980 (SEQ ID NO: 17).

[0179] DB1 / 162518159.2 24 ORN-109PC / 114384-5109

[0180] In an embodiment, the targeting moiety comprises AMP-514 (aka MEDI-0680).

[0181] In an embodiment, the targeting moiety comprises the PD-L2-Fc fusion protein AMP-224, which is disclosed in W02010 / 027827 and WO 2011 / 066342, the entire disclosures of which are hereby incorporated by reference. In such an embodiment, the targeting moiety may include a targeting domain which comprises SEQ ID NO:4 of W02010 / 027827 (SEQ ID NO :20) and / or the B7-DC fusion protein which comprises SEQ ID NO:83 of W02010 / 027827 (SEQ ID NO: 21).

[0182] In an embodiment, the targeting moiety comprises the peptide AUNP 12 or any of the other peptides disclosed in US 2011 / 0318373 or 8,907,053. For example, the targeting moiety may comprise AUNP 12 ( / .e., Compound 8 or SEQ ID NO:49 of US 2011 / 0318373) which has the sequence of:

[0183] SEQ ID NO: 22).

[0184] In an embodiment, the targeting moiety comprises the anti-PD-1 antibody 1 E3, or fragments thereof, as disclosed in US 2014 / 0044738, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 1 E3 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23; and / or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24

[0185] In an embodiment, the targeting moiety comprises the anti-PD-1 antibody 1 E8, or fragments thereof, as disclosed in US 2014 / 0044738, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 1 E8 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 25) and / or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 26

[0186] In an embodiment, the targeting moiety comprises the anti-PD-1 antibody 1 H3, or fragments thereof, as disclosed in US 2014 / 0044738, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 1 H3 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 27 and / or light chain variable region comprising the amino acid sequence of SEQ ID NO: 28

[0187] DBl / 162518159.2 25 ORN-109PC / 114384-5109

[0188] In an embodiment, the targeting moiety comprises a VHH directed against PD-1 as disclosed, for example, in US 8,907,065 and WO 2008 / 071447, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, the VHHs against PD-1 comprise SEQ ID NOS: 347-351 of US 8,907,065 (SEQ ID No: 347 of US 8,907,065 (SEQ ID NO: 29); SEQ ID No: 348 of US 8,907,065 (SEQ ID NQ:30); SEQ ID No: 349 of US 8,907,065 (SEQ ID NO:31); SEQ ID No: 350 of US 8,907,065 (SEQ ID NO:32); and SEQ ID No: 351 of US 8,907,065 (SEQ ID NO:33).

[0189] In an embodiment, the targeting moiety comprises any one of the anti-PD-1 antibodies, or fragments thereof, as disclosed in US2011 / 0271358 and WQ2010 / 036959, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID NOS: 25-29 of US2011 / 0271358 (SEQ ID No: 25 of US2011 / 0271358 (SEQ ID NO:34); SEQ ID No: 26 of US2011 / 0271358 (SEQ ID NO:35); SEQ ID No: 27 of US2011 / 0271358 (SEQ ID NO:36); SEQ ID No: 28 of US2011 / 0271358 (SEQ ID NO:37);and SEQ ID No: 29 of US2011 / 0271358 (SEQ ID NO:38));and / or a light chain comprising an amino acid sequence selected from SEQ ID NOS: 30-33 of US2011 / 0271358 (SEQ ID No: 30 of US2011 / 0271358(SEQ ID NO :39); SEQ ID No: 31 of US2011 / 0271358 (SEQ ID NQ:40); SEQ ID No: 32 of US2011 / 0271358 (SEQ ID NO:41); and SEQ ID No: 33 of US2011 / 0271358 (SEQ ID NO:42)).

[0190] In various embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes comprise one or more antibodies directed against PD-1 , or antibody fragments thereof, selected from TSR-042 (Tesaro, Inc.), REGN2810 (Regeneron Pharmaceuticals, Inc.), PDR001 (Novartis Pharmaceuticals), and BGB-A317 (BeiGene Ltd.)

[0191] In various embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have one or more targeting moieties directed against PD-L1. In some embodiments, the chimeric proteins or chimeric protein complexes have one or more targeting moieties, which selectively bind a PD- L1 polypeptide. In some embodiments, the chimeric proteins or chimeric protein complexes comprise one or more antibodies, antibody derivatives or formats, peptides or polypeptides, or fusion proteins that selectively bind a PD- L1 polypeptide.

[0192] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody MEDI4736 (aka durvalumab), or fragments thereof. MEDI4736 is selective for PD-L1 and blocks the binding of PD-L1 to the PD-1 and CD80 receptors. MEDI4736 and antigen-binding fragments thereof for use in the methods provided herein comprises a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. The sequence of MEDI4736 is disclosed in WQ / 2016 / 06272, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, MEDI4736 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising the amino acid sequence of (SEQ ID NO:43); and / or a light chain comprising the amino acid sequence of (SEQ ID NO:44).

[0193] DB1 / 162518159.2 26 ORN-109PC / 114384-5109

[0194] In illustrative embodiments, the M EDI4736 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4 of WO / 2016 / 06272 (SEQ ID NO:45); and / or a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 of WO / 2016 / 06272 (SEQ ID NO:46).

[0195] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody atezolizumab (aka MPDL3280A, RG7446), or fragments thereof. In illustrative embodiments, atezolizumab or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising the amino acid sequence of (SEQ ID NO:47); and / or a light chain comprising the amino acid sequence of (SEQ ID NO:48).

[0196] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody avelumab (aka MSB0010718C), or fragments thereof In illustrative embodiments, avelumab or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising the amino acid sequence of (SEQ ID NO:49); and / or a light chain comprising the amino acid sequence of (SEQ ID NQ:50).

[0197] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody BMS-936559 (aka 12A4, MDX-1105), or fragments thereof, as disclosed in US 2013 / 0309250 and WQ2007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, BMS-936559 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of: (SEQ ID NO:51); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO:52).

[0198] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 3G10, or fragments thereof, as disclosed in US 2013 / 0309250 and WQ2007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 3G10 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 53); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 54).

[0199] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 10A5, or fragments thereof, as disclosed in US 2013 / 0309250 and WQ2007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 10A5 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 55); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 56).

[0200] In an embodiment, the targeting moiety comprises the anti-PD-L 1 antibody 5F8, or fragments thereof, as disclosed in US 2013 / 0309250 and WQ2007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 5F8 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 57); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 58).

[0201] DB1 / 162518159.2 27 ORN-109PC / 114384-5109

[0202] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 10H10, or fragments thereof, as disclosed in US 2013 / 0309250 and WQ2007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 10H10 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 59); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 60).

[0203] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 1 B12, or fragments thereof, as disclosed in US 2013 / 0309250 and W02007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 1 B12 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 61); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 62).

[0204] In an embodiment, the targeting moiety comprises the an ti-PD-L 1 antibody 7H 1 , or fragments thereof, as disclosed in US 2013 / 0309250 and WQ2007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 7H1 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 63); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 64).

[0205] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 11 E6, or fragments thereof, as disclosed in US 2013 / 0309250 and WQ2007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 11 E6 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 65); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 66).

[0206] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 12B7, or fragments thereof, as disclosed in US 2013 / 0309250 and W02007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 12B7 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 67); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 68).

[0207] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 13G4, or fragments thereof, as disclosed in US 2013 / 0309250 and W02007 / 005874, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 13G4 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 69); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 70).

[0208] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 1 E12, or fragments thereof, as disclosed in US 2014 / 0044738, the entire disclosures of which are hereby incorporated by reference In illustrative embodiments, 1 E12 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 71); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 72).

[0209] DB1 / 162518159.2 28 ORN-109PC / 114384-5109

[0210] In an embodiment, the targeting moiety comprises the anti-PD-L 1 antibody 1 F4, or fragments thereof, as disclosed in US 2014 / 0044738, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 1 F4 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 73); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 74).

[0211] In an embodiment, the targeting moiety comprises the anti-PD-L 1 antibody 2G11 , or fragments thereof, as disclosed in US 2014 / 0044738, the entire disclosures of which are hereby incorporated by reference In illustrative embodiments, 2G11 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 75); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 76).

[0212] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 3B6, or fragments thereof, as disclosed in US 2014 / 0044738, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 3B6 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 77); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 78).

[0213] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 3D10, or fragments thereof, as disclosed in US 2014 / 0044738 and WO2012 / 145493, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 3D10 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of (SEQ ID NO: 79); and / or a light chain variable region comprising the amino acid sequence of (SEQ ID NO: 80).

[0214] In an embodiment, the targeting moiety comprises any one of the anti-PD-L1 antibodies disclosed in US2011 / 0271358 and WO2010 / 036959, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 34-38 of US2011 / 0271358 (SEQ ID No: 34 of US2011 / 0271358 (SEQ ID NO: 81); SEQ ID No: 35 of US2011 / 0271358 (SEQ ID NO: 82); SEQ ID No: 36 of US2011 / 0271358 (SEQ ID NO: 83); SEQ ID No: 37 of US2011 / 0271358 (SEQ ID NO: 84); and SEQ ID No: 38 of US2011 / 0271358 (SEQ ID NO: 85)); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 39-42 of US2011 / 0271358 (SEQ ID No: 39 of US2011 / 0271358 (SEQ ID NO: 86); SEQ ID No: 40 of US2011 / 0271358 (SEQ ID NO: 87); SEQ ID No: 41 of US2011 / 0271358 (SEQ ID NO: 88); and SEQ ID No: 42 of US2011 / 0271358(SEQ ID NO: 89)).

[0215] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 2.7A4, or fragments thereof, as disclosed in WO 2011 / 066389, US8,779,108, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 2.7A4 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID

[0216] DB1 / 162518159.2 29 ORN-109PC / 114384-5109

[0217] No: 2 of WO 2011 / 066389 (SEQ ID NO: 90); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 7 of WO 2011 / 066389 (SEQ ID NO: 91).

[0218] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 2.9D10, or fragments thereof, as disclosed in WO 2011 / 066389, US8,779,108, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 2.9D10 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No: 12 of WO 2011 / 066389 (SEQ ID NO: 92); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 17 of WO 2011 / 066389 (SEQ ID NO: 93).

[0219] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 2.14H9, or fragments thereof, as disclosed in WO 2011 / 066389, US8,779,108, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 2.14H9 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No: 22 of WO 2011 / 066389 (SEQ ID NO: 94); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 27 of WO 2011 / 066389 (SEQ ID NO: 95).

[0220] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 2.20A8, or fragments thereof, as disclosed in WO 2011 / 066389, US8,779,108, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 2.20A8 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No: 32 of WO 2011 / 066389 (SEQ ID NO: 96); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 37 of WO 2011 / 066389 (SEQ ID NO: 97).

[0221] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 3.15G8, or fragments thereof, as disclosed in WO 2011 / 066389, US8,779,108, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 3.15G8 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No: 42 of WO 2011 / 066389 (SEQ ID NO: 98); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 47 of WO 2011 / 066389 (SEQ ID NO: 99).

[0222] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 3.18G1 , or fragments thereof, as disclosed in WO 2011 / 066389, US8,779,108, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 3.18G1 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No: 52 of WO 2011 / 066389 (SEQ ID NO: 100); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 57 of WO 2011 / 066389 (SEQ ID NO: 101).

[0223] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 2.7A4OPT, or fragments thereof, as disclosed in WO 2011 / 066389, US8.779, 108, and US2014 / 0356353, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 2 7A4OPT or an antigen-binding

[0224] DB1 / 162518159.2 30 ORN-109PC / 114384-5109 fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No: 62 of WO 2011 / 066389 (SEQ ID NO:102); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 67 of WO 2011 / 066389 (SEQ ID NO: 103).

[0225] In an embodiment, the targeting moiety comprises the anti-PD-L1 antibody 2.14H9OPT, or fragments thereof, as disclosed in WO 2011 / 066389, US8,779,108, and US2014 / 0356353, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, 2.14H9OPT or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No: 72 of WO 2011 / 066389 (SEQ ID NO: 104); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 77 of WO 2011 / 066389 (SEQ ID NO: 105).

[0226] In an embodiment, the targeting moiety comprises any one of the anti-PD-L1 antibodies disclosed in W02016 / 061142, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 18, 30, 38, 46, 50, 54, 62, 70, and 78 of WQ2016 / 061142 (SEQ ID No: 18 of WQ2016 / 061142 (SEQ ID NQ:106); SEQ ID No: 30 of WQ2016 / 061142 (SEQ ID NQ:107); SEQ ID No: 38 of WQ2016 / 061142 (SEQ ID NO:108); SEQ ID No: 46 of WQ2016 / 061142 (SEQ ID NO:109); SEQ ID No: 50 of WQ2016 / 061142 (SEQ ID NQ:110); SEQ ID No: 54 of WQ2016 / 061142 (SEQ ID NO:111); SEQ ID No: 62 of WQ2016 / 061142 (SEQ ID NO:112); SEQ ID No: 70 of WQ2016 / 061142 (SEQ ID NO:113); and SEQ ID No: 78 of WQ2016 / 061142 (SEQ ID NO:114)); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 22, 26, 34, 42, 58, 66, 74, 82, and 86 of WQ2016 / 061142 (SEQ ID No: 22 of WQ2016 / 061142 (SEQ ID NO:115); SEQ ID No: 26 of WQ2016 / 061142 (SEQ ID NO:116); SEQ ID No: 34 of WQ2016 / 061142 (SEQ ID NO: 117); SEQ ID No: 42 of WQ2016 / 061142 (SEQ ID NO:118);SEQ ID No: 58 of WQ2016 / 061142 (SEQ ID NO:119); SEQ ID No: 66 of WQ2016 / 061142(SEQ ID NQ:120); SEQ ID No: 74 of WQ2016 / 061142 (SEQ ID NO: 121 ); SEQ ID No: 82 of WQ2016 / 061142 (SEQ ID NO: 122); and SEQ ID No: 86 of WQ2016 / 061142 (SEQ ID NO:123)).

[0227] In an embodiment, the targeting moiety comprises any one of the anti-PD-L1 antibodies disclosed in WQ2016 / 022630, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, and 46 of WQ2016 / 022630 ( SEQ ID No: 2 of WQ2016 / 022630 (SEQ ID NO: 124); SEQ ID No: 6 of WQ2016 / 022630 (SEQ ID NO: 125); SEQ ID No: 10 of WQ2016 / 022630 (SEQ ID NO: 126); SEQ ID No: 14 of WQ2016 / 022630 (SEQ ID NO:127); SEQ ID No: 18 of WQ2016 / 022630 (SEQ ID NO:128); SEQ ID No: 22 of WQ2016 / 022630 (SEQ ID NO:129); SEQ ID No: 26 of WQ2016 / 022630 (SEQ ID NO: 130); SEQ ID No: 30 of WQ2016 / 022630 (SEQ ID NO:131); SEQ ID No: 34 of WQ2016 / 022630 (SEQ ID NO: 132); SEQ ID No: 38 of WQ2016 / 022630 (SEQ ID NO:133); SEQ ID No: 42 of WQ2016 / 022630 (SEQ ID NO: 134); and SEQ ID No: 46 of WQ2016 / 022630 (SEQ ID NO: 135)); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, and 48 of WQ2016 / 022630 (SEQ ID No: 4 of WQ2016 / 022630 (SEQ ID NO: 136); SEQ ID No:

[0228] DB1 / 162518159.2 31 ORN-109PC / 114384-5109

[0229] 8 of W02016 / 022630 (SEQ ID NO:137); SEQ ID No: 12 of W02016 / 022630 (SEQ ID NO:138); SEQ ID No: 16 of W02016 / 022630 (SEQ ID NO:139); SEQ ID No: 20 of WQ2016 / 022630 (SEQ ID NQ: 140); SEQ ID No: 24 of

[0230] WQ2016 / 022630 (SEQ ID NO:141); SEQ ID No: 28 of WQ2016 / 022630 (SEQ ID NO: 142); SEQ ID No: 32 of

[0231] WQ2016 / 022630 (SEQ ID NO:143); SEQ ID No: 36 of WQ2016 / 022630 (SEQ ID NO: 144); SEQ ID No: 40 of

[0232] WQ2016 / 022630 (SEQ ID NO: 145); SEQ ID No: 44 of WQ2016 / 022630 (SEQ ID NO: 146); and SEQ ID No: 48 of

[0233] WQ2016 / 022630 (SEQ ID NO: 147)).

[0234] In an embodiment, the targeting moiety comprises any one of the anti-PD-L1 antibodies disclosed in WO2015 / 112900, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 38, 50, 82, and 86 of WO 2015 / 112900 (SEQ ID No: 38 of WQ2015 / 112900 (SEQ ID NO:148); SEQ ID No: 50 of WO 2015 / 112900 (SEQ ID NO:149); SEQ ID No: 82 of WO 2015 / 112900 (SEQ ID NO: 150); and SEQ ID No: 86 of WO 2015 / 112900 (SEQ ID NO:151 )); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 42, 46, 54, 58, 62, 66, 70, 74, and 78 of WO 2015 / 112900 (SEQ ID No: 42 of WO2015 / 112900 (SEQ ID NO:152); SEQ ID No: 46 of WO 2015 / 112900: (SEQ ID NO:153); SEQ ID No: 54 of WO 2015 / 112900 (SEQ ID NO:154); SEQ ID No: 58 of WO

[0235] 2015 / 112900 (SEQ ID NO:155); SEQ ID No: 62 of WO 2015 / 112900 (SEQ ID NO:156); SEQ ID No: 66 of WO

[0236] 2015 / 112900 (SEQ ID NO:157); SEQ ID No: 70 of WO 2015 / 112900 (SEQ ID NO:158); SEQ ID No: 74 of WO

[0237] 2015 / 112900 (SEQ ID NO:159); and SEQ ID No: 78 of WO 2015 / 112900 (SEQ ID NQ:160)).

[0238] In an embodiment, the targeting moiety comprises any one of the anti-PD-L1 antibodies disclosed in WO 2010 / 077634 and US 8,217,149, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, the anti-PD-L1 antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain region comprising the amino acid sequence of SEQ ID No: 20 of WO 2010 / 077634 (SEQ ID NO: 161); and / or a light chain variable region comprising the amino acid sequence of SEQ ID No: 21 of WO 2010 / 077634 (SEQ ID NO: 162)

[0239] In an embodiment, the targeting moiety comprises any one of the anti-PD-L1 antibodies obtainable from the hybridoma accessible under CNCM deposit numbers CNCM 1-4122, CNCM I-4080 and CNCM 1-4081 as disclosed in US 20120039906, the entire disclosures of which are hereby incorporated by reference.

[0240] In an embodiment, the targeting moiety comprises a VHH directed against PD-L1 as disclosed, for example, in US 8,907,065 and WO 2008 / 071447, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, the VHHs against PD-L1 comprise SEQ ID NOS: 394-399 of US 8,907,065 (SEQ ID No: 394 of US 8,907,065 (SEQ ID NO: 163); SEQ ID No: 395 of US 8,907,065 (SEQ ID NOU 64); SEQ ID No: 396 of US 8,907,065 (SEQ ID NO: 165); SEQ ID No: 397 of US 8,907,065 (SEQ ID NO:166); SEQ ID No: 398 of US 8,907,065 (SEQ ID NO: 167); and SEQ ID No: 399 of US 8,907,065 (SEQ ID NO: 168)).

[0241] In various embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have one or more targeting moieties directed against PD-L2. In some embodiments, the

[0242] DB1 / 162518159.2 32 ORN-109PC / 114384-5109 chimeric proteins or chimeric protein complexes have one or more targeting moieties which selectively bind a PD- L2 polypeptide. In some embodiments, the chimeric proteins or chimeric protein complexes comprise one or more antibodies, antibody derivatives or formats, peptides or polypeptides, or fusion proteins that selectively bind a PD- L2 polypeptide.

[0243] In an embodiment, the targeting moiety comprises a VHH directed against PD-L2 as disclosed, for example, in US 8,907,065 and WO 2008 / 071447, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, the VHHs against PD-L2 comprise SEQ ID Nos: 449-455 of US 8,907,065 (SEQ ID No: 449 of US 8,907,065 (SEQ ID NO: 169); SEQ ID No: 450 of US 8,907,065 (SEQ ID NQ:170); SEQ ID No: 451 of US 8,907,065 (SEQ ID NO:171); SEQ ID No: 452 of US 8,907,065 (SEQ ID NO:172); SEQ ID No: 453 of US 8,907,065 (SEQ ID NO: 173); SEQ ID No: 454 of US 8,907,065 (SEQ ID NO: 174); and SEQ ID No: 455 of US 8,907,065 (SEQ ID NO:175)).

[0244] In an embodiment, the targeting moiety comprises any one of the anti-PD-L2 antibodies disclosed in US2011 / 0271358 and WQ2010 / 036959, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 43-47 of US2011 / 0271358 (SEQ ID No: 43 of US2011 / 0271358 (SEQ ID NO:176); SEQ ID No: 44 of US2011 / 0271358 (SEQ ID NO:177); SEQ ID No: 45 of US2011 / 0271358 (SEQ ID NO:178); SEQ ID No: 46 of US2011 / 0271358 (SEQ ID NO: 179); and SEQ ID No: 47 of US2011 / 0271358 (SEQ ID NO: 180)); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 48-51 of US2011 / 0271358 (SEQ ID No: 48 of US2011 / 0271358 (SEQ ID NO:181); SEQ ID No: 49 of US2011 / 0271358 (SEQ ID NO:182); SEQ ID No: 50 of US2011 / 0271358 (SEQ ID NO: 183); and SEQ ID No: 51 of US2011 / 0271358 (SEQ ID NO:184)).

[0245] In various embodiments, the targeting moieties of the invention may comprise a sequence that targets PD-1 , PD- L1 , and / or PD-L2 which is at least about 60%, at least about 61 %, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any of the sequences disclosed herein (e.g. about 60%, or about 61%, or about 62%, or about 63%, or about 64%, or about 65%, or about 66%, or about 67%, or about 68%, or about 69%, or about 70%, or about 71%, or about 72%, or about 73%, or about 74%, or about 75%, or about 76%, or about 77%, or about 78%, or about 79%, or about 80%, or about 81%, or about 82%, or about 83%, or about 84%, or about 85%, or about 86%, or about 87%, or about 88%, or about 89%, or about 90%, or about 91 %, or about 92%, or about 93%, or about 94%, or about 95%, or about 96%, or about 97%, or about 98%, about 99% or about 100% sequence identity with any of the sequences disclosed herein).

[0246] DB1 / 162518159.2 33 ORN-109PC / 114384-5109

[0247] In various embodiments, the targeting moieties of the invention may comprise any combination of heavy chain, light chain, heavy chain variable region, light chain variable region, complementarity-determining region (CDR), and framework region sequences that target PD-1 , PD-L1, and / or PD-L2 as disclosed herein.

[0248] Additional antibodies, antibody derivatives or formats, peptides or polypeptides, or fusion proteins that selectively bind or target PD-1, PD-L1 and / or PD-L2 are disclosed in WO 2011 / 066389, US 2008 / 0025980, US 2013 / 0034559, US 8,779,108, US 2014 / 0356353, US 8,609,089, US 2010 / 028330, US 2012 / 0114649, WO 2010 / 027827, WO 2011. / 066342, US 8,907,065, WO 2016 / 062722, WO 2009 / 101611 , WO2010 / 027827, WO 2011 / 066342, WO 2007 / 005874 , WO 2001 / 014556, US2011 / 0271358, WO 2010 / 036959, WO 2010 / 077634, US 8,217,149, US 2012 / 0039906, WO 2012 / 145493, US 2011 / 0318373, U.S. Patent No. 8,779,108, US 20140044738, WO 2009 / 089149, WO 2007 / 00587, WO 2016061142, WO 2016,02263, WO 2010 / 077634, and WO 2015 / 112900, the entire disclosures of which are hereby incorporated by reference.

[0249] In one embodiment, the present chimeric proteins or chimeric protein complexes have (i) a targeting moiety directed against a T cell, for example, mediated by targeting to CD8 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein In an embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have a targeting moiety directed against CD8 on T cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0250] In one embodiment, the present chimeric proteins or chimeric protein complexes have (i) a targeting moiety directed against a T cell, for example, mediated by targeting to CD4 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein In an embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have a targeting moiety directed against CD4 on T cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0251] In one embodiment, the present chimeric proteins or chimeric protein complexes have (i) a targeting moiety directed against a T cell, for example, mediated by targeting to CD3, CXCR3, CCR4, CCR9, CD70, CD103, or one or more immune checkpoint markers and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have a targeting moiety directed against CD3 on T cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0252] In some embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complexes have one or more targeting moieties directed against CD3 expressed on T cells. In some embodiments, the chimeric protein or chimeric protein complex have one or more targeting moieties, which selectively bind a CD3 polypeptide. In some embodiments, the chimeric protein or chimeric protein complex comprise one or more antibodies, antibody derivatives or formats, peptides or polypeptides, or fusion proteins that selectively bind a CD3 polypeptide.

[0253] DB1 / 162518159.2 34 ORN-109PC / 114384-5109

[0254] In an embodiment, the targeting moiety comprises the anti-CD3 antibody muromonab-CD3 (aka Orthoclone OKT3), or fragments thereof. Muromonab-CD3 is disclosed in U.S. Patent No. 4,361,549 and Wilde et al. (1996) 51 :865-894, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, muromonab-CD3 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising the amino acid sequence of (SEQ ID NO:185); and / or a light chain comprising the amino acid sequence of (SEQ ID NO: 186).

[0255] In an embodiment, the targeting moiety comprises the anti-CD3 antibody otelixizumab, or fragments thereof. Otelixizumab is disclosed in U.S Patent Publication No. 20160000916 and Chatenoud et al. (2012) 9:372-381, the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, otelixizumab or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising the amino acid sequence of: SEQ ID NO: 187; and / or a light chain comprising the amino acid sequence of SEQ ID NO:188.

[0256] In an embodiment, the targeting moiety comprises the anti-CD3 antibody teplizumab (AKA MGA031 and hOKT3y1 (Ala-Ala)), or fragments thereof. Teplizumab is disclosed in Chatenoud et al. (2012) 9:372-381 , the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, teplizumab or an antigenbinding fragment thereof for use in the methods provided herein comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:189; and / or a light chain comprising the amino acid sequence of SEQ ID NQ:190.

[0257] In an embodiment, the targeting moiety comprises the anti-CD3 antibody visilizumab (AKA Nuvion®; HuM291), or fragments thereof. Visilizumab is disclosed in U.S. 5,834,597 and WQ2004052397, and Cole eta / ., Transplantation (1999) 68:563-571 , the entire disclosures of which are hereby incorporated by reference. In illustrative embodiments, visilizumab or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:191 ; and / or a light chain variable region comprising the amino acid sequence of SEQ ID NO:192.

[0258] In an embodiment, the targeting moiety comprises the anti-CD3 antibody foralumab (aka N 1-0401 ), or fragments thereof. In various embodiments, the targeting moiety comprises any one of the anti-CD3 antibodies disclosed in US20140193399, US 7,728,114, US20100183554, and US 8,551 ,478, the entire disclosures of which are hereby incorporated by reference.

[0259] In illustrative embodiments, the anti-CD3 antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID Nos: 2 and 6 of US 7,728,114 (SEQ ID No: 2 of US 7,728,114 (SEQ ID NO:193) and SEQ ID No: 6 of US 7,728,114 (SEQ ID NO: 194)); and / or a light chain variable region comprising the amino acid sequence of SEQ ID NOs 4 and 8 of US 7,728,114 (SEQ ID No: 4 of US 7,728,114 (SEQ ID NO:195) and SEQ ID No: 8 of US 7,728,114 (SEQ ID NO:196)).

[0260] In an embodiment, the targeting moiety comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2 of US 7,728,114 and a light chain variable region comprising the amino acid sequence

[0261] DB1 / 162518159.2 35 ORN-109PC / 114384-5109 of SEQ ID NO:4 of US 7,728,114. In an embodiment, the targeting moiety comprises any one of the anti-CD3 antibodies disclosed in US2016 / 0168247, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 6-9 of US2016 / 0168247 (SEQ ID No: 6 of US2016 / 0168247 (SEQ ID NO:197); SEQ ID No: 7 of US2016 / 0168247 (SEQ ID NO: 198); SEQ ID No: 8 of US2016 / 0168247 (SEQ ID NO:199); and SEQ ID No: 9 of US2016 / 0168247 (SEQ ID NQ:200)); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 10-12 of US2016 / 0168247 (SEQ ID No: 10 of US2016 / 0168247 (SEQ ID NQ:201); SEQ ID No: 11 of US2016 / 0168247 (SEQ ID NQ:202); and SEQ ID No: 12 of US2016 / 0168247 (SEQ ID NQ:203)).

[0262] In an embodiment, the targeting moiety comprises any one of the anti-CD3 antibodies disclosed in US2015 / 0175699, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID No: 9 of US2015 / 0175699 (SEQ ID NQ:204); and / or a light chain comprising an amino acid sequence selected from SEQ ID No: 10 of US2015 / 0175699 (SEQ ID NQ:205).

[0263] In an embodiment, the targeting moiety comprises any one of the anti-CD3 antibodies disclosed in US 8,784,821 , the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 2, 18, 34, 50, 66, 82, 98 and 114 of US 8,784,821 (SEQ ID No: 2 of US 8,784,821 (SEQ ID NQ:206); SEQ ID No: 18 of US 8,784,821 (SEQ ID NQ:207); SEQ ID No: 34 of US 8,784,821 (SEQ ID NQ:208); SEQ ID No: 50 of US 8,784,821 (SEQ ID NQ:209); SEQ ID No: 66 of US 8,784,821 (SEQ ID NQ:210); SEQ ID No: 82 of US 8,784,821 (SEQ ID NO:211); SEQ ID No: 98 of US 8,784,821 (SEQ ID NO:212); and SEQ ID No: 114 of US 8,784,821 (SEQ ID NO:213)); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 10, 26, 42, 58, 74, 90, 106 and 122 of US 8,784,821 (SEQ ID No: 10 of US 8,784,821 (SEQ ID NO:214); SEQ ID No: 26 of US 8,784,821 (SEQ ID NO:215); SEQ ID No: 42 of US 8,784,821 (SEQ ID NO:216); SEQ ID No: 58 of US 8,784,821 (SEQ ID NO:217); SEQ ID No: 74 of US 8,784,821 (SEQ ID NO:218); SEQ ID No: 90 of US 8,784,821 (SEQ ID NO:219); SEQ ID No: 106 of US 8,784,821 (SEQ ID NQ:220); and SEQ ID No: 122 of US 8,784,821 (SEQ ID NO:221).

[0264] In an embodiment, the targeting moiety comprises any one of the anti-CD3 binding constructs disclosed in US20150118252, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 6 and 86 of US20150118252 (SEQ ID No: 6 of US20150118252 (SEQ ID NO :222) and SEQ ID No: 86 of US20150118252 (SEQ ID NO:223)) and / or a light chain comprising an amino acid sequence selected from SEQ ID No: 3 of US2015 / 0175699 (SEQ ID No: 3 of US20150118252 (SEQ ID NO:224)).

[0265] DB1 / 162518159.2 36 ORN-109PC / 114384-5109

[0266] In an embodiment, the targeting moiety comprises any one of the anti-CD3 binding proteins disclosed in US2016 / 0039934, the entire contents of which are hereby incorporated by reference. In illustrative embodiments, the antibody or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain comprising an amino acid sequence selected from SEQ ID Nos: 6-9 of US2016 / 0039934 (SEQ ID No: 6 of US2016 / 0039934 (SEQ ID NO:225); SEQ ID No: 7 of US2016 / 0039934 (SEQ ID NO:226); SEQ ID No: 8 of US2016 / 0039934 (SEQ ID NO:227); and SEQ ID No: 9 of US2016 / 0039934 (SEQ ID NO:228)); and / or a light chain comprising an amino acid sequence selected from SEQ ID Nos: 1-4 of US2016 / 0039934 (SEQ ID No: 1 of US2016 / 0039934 (SEQ ID NO:229); SEQ ID No: 2 of US2016 / 0039934 (SEQ ID NQ:230); SEQ ID No: 3 of US2016 / 0039934 (SEQ ID NO:231); and SEQ ID No: 4 of US2016 / 0039934 (SEQ ID NO:232)).

[0267] In various embodiments, the targeting moieties of the invention may comprise a sequence that targets CD3 which is at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71 %, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any of the sequences disclosed herein (e.g. about 60%, or about 61 %, or about 62%, or about 63%, or about 64%, or about 65%, or about 66%, or about 67%, or about 68%, or about 69%, or about 70%, or about 71%, or about 72%, or about 73%, or about 74%, or about 75%, or about 76%, or about 77%, or about 78%, or about 79%, or about 80%, or about 81 %, or about 82%, or about 83%, or about 84%, or about 85%, or about 86%, or about 87%, or about 88%, or about 89%, or about 90%, or about 91%, or about 92%, or about 93%, or about 94%, or about 95%, or about 96%, or about 97%, or about 98%, about 99% or about 100% sequence identity with any of the sequences disclosed herein)

[0268] In various embodiments, the targeting moieties of the invention may comprise any combination of heavy chain, light chain, heavy chain variable region, light chain variable region, complementarity determining region (CDR), and framework region sequences that target CD3 as disclosed herein. In various embodiments, the targeting moieties of the invention may comprise any heavy chain, light chain, heavy chain variable region, light chain variable region, complementarity determining region (CDR), and framework region sequences of the CD3-specific antibodies including, but not limited to, X35-3, VIT3, BMA030 (BW264 / 56), CLB-T3 / 3, CRIS7, YTH12.5, Fl 11-409, CLB-T3.4.2, TR-66, WT32, SPv-T3b, 11 D8, XIII-141, XIII-46, XIII-87, 12F6, T3 / RW2-8C8, T3 / RW2-4B6, OKT3D, M-T301, SMC2, WT31 and F101 01. These CD3-specific antibodies are well known in the art and, inter alia, described in Tunnacliffe (1989), Int. Immunol. 1 , 546-550, the entire disclosures of which are hereby incorporated by reference.

[0269] Additional antibodies, antibody derivatives or formats, peptides or polypeptides, or fusion proteins that selectively bind or target CD3 are disclosed in US Patent Publication No. 2016 / 0000916, US Patent Nos. 4,361,549,

[0270] DB1 / 162518159.2 37 ORN-109PC / 114384-5109

[0271] 5,834,597, 6,491,916, 6,406,696, 6,143,297, 6,750,325 and International Publication No. WO 2004 / 052397, the entire disclosures of which are hereby incorporated by reference.

[0272] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a T cell, for example, mediated by targeting to PD-1 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0273] By way of non-limiting example, in various embodiments, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a B cell, for example, mediated by targeting to CD10, CD19, CD20, CD21 , CD22, CD23, CD24, CD37, CD38, CD39, CD40, CD70, CD72, CD73, CD74, CDw75, CDw76, CD77, CD78, CD79a / b, CD80, CD81 , CD82, CD83, CD84, CD85, CD86, CD89, CD98, CD126, CD127, CDw130, CD138, or CDw150; and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric proteins or chimeric protein complexes have a targeting moiety directed against CD20.

[0274] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a B cell, for example, mediated by targeting to CD19, CD20 or CD70 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0275] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a B cell, for example, mediated by targeting to CD20 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric proteins or chimeric protein complexes have a targeting moiety directed against CD20 on B cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells. By way of example, in some embodiments, the CD20 targeting moiety is a recombinant heavy-chain-only antibody (VHH) having the sequence of:

[0276] QVQLQESGGGLAQAGGSLRLSCAASGRTFSMGWFRQAPGKEREFVAAITYSGGSPYYASSVRGRFTISRDNAK NTVYLQMNSLKPEDTAVYYCAANPTYGSDWNAENWGQGTQVTVSS (SEQ ID NO: 288).

[0277] By way of non-limiting example, in various embodiments, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a NK cell, for example, mediated by targeting to 2B4 / SLAMF4, KIR2DS4, CD155 / PVR, KIR3DL1 , CD94, LMIR1 / CD300A, CD69, LMIR2 / CD300c, CRACC / SLAMF7, LMIR3 / CD300LF, DNAM-1 , LMIR5 / CD300LB, Fc-epsilon RII, LMIR6 / CD300LE, Fc-y RI / CD64, MICA, Fc-y Rl I B / CD32b, MICB, Fc-y RIIC / CD32c, MULT-1, Fc-y RIIA / CD32a, Nectin-2 / CD112, Fc-y RIII / CD16, NKG2A, FcRH1 / IRTA5, NKG2C, FcRH2 / IRTA4, NKG2D, FcRH4 / IRTA1 , NKp30, FcRH5 / IRTA2, NKp44, Fc-Receptor-like 3 / CD16-2, NKp46 / NCR1, NKp80 / KLRF1, NTB-A / SLAMF6, Rae-1 , Rae-1 a, Rae-1 [3, Rae-1 delta, H60, Rae-1 epsilon, ILT2 / CD85j, Rae-1 y, ILT3 / CD85k, TREM-1, ILT4 / CD85d, TREM-2, ILT5 / CD85a, TREM-3, KIR / CD158, TREML1 / TLT-1 , KIR2DL1 , ULBP-1 , KIR2DL3, ULBP-2, KIR2DL4 / CD 158d, or ULBP-3; and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0278] DB1 / 162518159.2 38 ORN-109PC / 114384-5109

[0279] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a NK cell, for example, mediated by targeting to Kiri alpha, DNAM-1 or CD64 and (ii) a targeting moiety is directed against a tumor cell, along with any of the signaling agents (e.g., IL-6 or variant thereof) described herein.

[0280] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a NK cell, for example, mediated by targeting to KIR1 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric proteins or chimeric protein complexes have a targeting moiety directed against KIR1 on NK cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0281] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a NK cell, for example, mediated by targeting to TIGIT or KIR1 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric proteins or chimeric protein complexes have a targeting moiety directed against TIGIT on NK cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0282] By way of non-limiting example, in various embodiments, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a dendritic cell, for example, mediated by targeting to CLEC-9A, XCR1 , RANK, CD36 / SRB3, LOX-1 / SR-E1, CD68, MARCO, CD163, SR-A1 / MSR, CD5L, SREC-1, CL-PI / COLEC 12, SREC-II, LIMPIIISRB2, RP105, TLR4, TLR1 , TLR5, TLR2, TLR6, TLR3, TLR9, 4-IBB Ligand / TN FSF9, IL-12 / IL-23 p40, 4- Amino-1,8-naphthalimide, ILT2 / CD85j, CCL21 / 6Ckine, ILT3 / CD85k, 8-oxo-dG, ILT4 / CD85d, 8D6A, ILT5 / CD85a, A2B5, lutegrin a 4 / CD49d, Aag, Integrin 2 / CD18, AMICA, Langerin, B7-2 / CD86, Leukotriene B4 Rl, B7-H3, LMIR1 / CD300A, BLAME / SLAMF8, LM I R2 / CD300c, Clq R1 / CD93, LMIR3 / CD300LF, CCR6, LMIR5 / CD300LB CCR7, LMIR6 / CD300LE, CD40 / TNFRSF5, MAG / Siglec-4-a, CD43, MCAM, CD45, MD-1, CD68, MD-2, CD83, MDL-1 / CLEC5A, CD84 / SLAMF5, MMR, CD97, NCAMLI, CD2F-10 / SLAMF9, Osteoactivin GPNMB, Chern 23, PD- L2, CLEC-1 , RP105, CLEC-2, Siglec-2 / CD22, CRACC / SLAMF7, Siglec-3 / CD33, DC-SIGN, Siglec-5, DC- SIGNR / CD299, Siglec-6, DCAR, Siglec-7, DCIR / CLEC4A, Siglec-9, DEC-205, Siglec-10, Dectin-1 / CLEC7A, Siglec-F, Dectin-2 / CLEC6A, SIGNR1 / CD209, DEP-1 / CD148, SIGNR4, DLEC, SLAM, EMMPRIN / CD147, TCCR / WSX-1, Fc-y R1 / CD64, TLR3, Fc-y RIIB / CD32b, TREM-1, Fc-y RIIC / CD32c, TREM-2, Fc-y RIIA / CD32a, TREM-3, Fc-y RIII / CD16, TREML1 / TLT-1, ICAM-2 / CD102, or Vanilloid R1 ; and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0283] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a dendritic cell, for example, mediated by targeting to CLEC-9A, DC-SIGN, CD64, CLEC4A, or DEC205 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric protein or chimeric protein complex have a targeting moiety directed against CLEC9A on dendritic cells and a second targeting moiety directed against PD-L1 or PD- L2 on tumor cells.

[0284] DB1 / 162518159.2 39 ORN-109PC / 114384-5109

[0285] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a dendritic cell, for example, mediated by targeting to CLEC9A and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric proteins or chimeric protein complexes have a targeting moiety directed against CLEC9A on dendritic cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0286] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a dendritic cell, for example, mediated by targeting to XCR1 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric proteins or chimeric protein complexes have a targeting moiety directed against XCR1 on dendritic cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0287] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a dendritic cell, for example, mediated by targeting to RANK and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric protein or chimeric protein complex have a targeting moiety directed against RANK on dendritic cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0288] By way of non-limiting example, in various embodiments, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a monocyte / macrophage, for example, mediated by targeting to SIRP1 a, B7- 1 / CD80, ILT4 / CD85d, B7-H1 , ILT5 / CD85a, Common p Chain, Integrin a 4 / CD49d, BLAME / SLAMF8, Integrin a X / CDIIc, CCL6 / C10, Integrin p 2 / CD18, CD155 / PVR, Integrin p 3 / CD61, CD31 / PECAM-1 , Latexin, CD36 / SR-B3, Leukotriene B4 R1 , CD40 / TNFRSF5, LIMPIIISR-B2, CD43, LMIR1 / CD300A, CD45, LMIR2 / CD300c, CD68, LMIR3 / CD300LF, CD84 / SLAMF5, LMIR5 / CD300LB, CD97, LMIR6 / CD300LE, CD163, LRP-1 , CD2F-10 / SLAMF9, MARCO, CRACC / SLAMF7, MD-1, ECF-L, MD-2, EMMPRIN / CD147, MGL2, Endoglin / CD105, Osteoactivin / GPNMB, Fc-y RI / CD64, Osteopontin, Fc-y Rl I B / CD32b, PD-L2, Fc-y RIIC / CD32c, Siglec-3 / CD33, Fc- y RIIA / CD32a, SIGNR1 / CD209, Fc-y RIII / CD16, SLAM, GM-CSF R a, TCCR / WSX-1, ICAM-2 / CD102, TLR3, IFN- y Rl, TLR4, IFN- y R2, TREM-I, IL-I RII, TREM-2, ILT2 / CD85j, TREM-3, ILT3 / CD85k, TREML1 / TLT-1 , 2B4 / SLAMF 4, IL-10 R a, ALCAM, IL-10 R p, AminopeptidaseN / ANPEP, ILT2 / CD85j, Common p Chain, ILT3 / CD85k, Clq R1 / CD93, ILT4 / CD85d, CCR1, ILT5 / CD85a, CCR2, CD206, Integrin a 4 / CD49d, CCR5, Integrin a M / CDII b, CCR8, Integrin a X / CDIIc, CD155 / PVR, Integrin p 2 / CD18, CD14, Integrin p 3 / CD61 , CD36 / SR-B3, LAIR1, CD43, LAIR2, CD45, Leukotriene B4-R1, CD68, LIMPIIISR-B2, CD84 / SLAMF5, LMIR1 / CD300A, CD97, LMIR2 / CD300c, CD163, LMIR3 / CD300LF, Coagulation Factor lll / Tissue Factor, LMIR5 / CD300LB, CX3CR1 , CX3CL1 , LMIR6 / CD300LE, CXCR4, LRP-1, CXCR6, M-CSF R, DEP-1 / CD148, MD-1 , DNAM-1 , MD-2, EMMPRIN / CD147, MMR, Endoglin / CD105, NCAM-L1, FC-Y RI / CD64, PSGL-1, Fc-y RIIIICD16, RP105, G-CSF R, L-Selectin, GM-CSF R a, Siglec-3 / CD33, HVEM / TNFRSF14, SLAM, ICAM-1 / CD54, TCCR / WSX-1, ICAM-2 / CD102, TREM-I, IL-6 R, TREM- 2, CXCRI / I L-8 RA, TREM-3, or TREMLI / TLT-1 ; and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0289] DB1 / 162518159.2 40 ORN-109PC / 114384-5109

[0290] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a monocyte / macrophage, for example, mediated by targeting to B7-H1, CD31 / PECAM-1, CD163, CCR2, or Macrophage Mannose Receptor CD206 and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0291] In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a monocyte / macrophage, for example, mediated by targeting to SIRP1 a and (ii) a targeting moiety is directed against a tumor cell, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein. In an embodiment, the chimeric protein or chimeric protein complex have a targeting moiety directed against SIRP1 a on macrophage cells and a second targeting moiety directed against PD-L1 or PD-L2 on tumor cells.

[0292] In various embodiments, the chimeric protein or chimeric protein complex have one or more targeting moieties directed against a checkpoint marker, e.g. one or more of PD-1 / PD-L1 or PD-L2, CD28 / CD80 or CD86, CTLA4 / CD80 or CD86, ICOS / ICOSL or B7RP1 , BTLA / HVEM, KIR, LAG3, CD137 / CD137L, OX40 / OX40L, CD27, CD40L, TIM3 / Gal9, CD47, CD70, and A2aR. In one embodiment, the chimeric protein or chimeric protein complex have (i) a targeting moiety directed against a checkpoint marker on a T cell, for example, PD-1 and (ii) a targeting moiety directed against a tumor cell, for example, PD-L1 or PD-L2, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein In an embodiment, the chimeric protein or chimeric protein complex have a targeting moiety directed against PD-1 on T cells and a second targeting moiety directed against PD-L1 on tumor cells. In another embodiment, the chimeric protein or chimeric protein complex have a targeting moiety directed against PD-1 on T cells and a second targeting moiety directed against PD-L2 on tumor cells.

[0293] In some embodiments, the chimeric protein or chimeric protein complex comprises two or more targeting moieties directed to the same or different immune cells. In some embodiments the chimeric protein or chimeric protein complex have (i) one or more targeting moieties directed against an immune cell selected from a T cell, a B cell, a dendritic cell, a macrophage, a NK cell, or subsets thereof and (ii) one or more targeting moieties directed against either the same or another immune cell selected from a T cell, a B cell, a dendritic cell, a macrophage, a NK cell, or subsets thereof, along with any of the antagonistic signaling agents (e.g., IL-6 antagonist) described herein.

[0294] In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a T cell and one or more targeting moieties directed against the same or another T cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a T cell and one or more targeting moieties directed against a B cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a T cell and one or more targeting moieties directed against a dendritic cell In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against a T cell and one or more targeting moieties directed against a macrophage. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against a T cell and one or more targeting moieties directed against a NK cell. For example, in an illustrative embodiment, the chimeric protein or chimeric protein complex may include a targeting

[0295] DB1 / 162518159.2 41 ORN-109PC / 114384-5109 moiety against CD8 and a targeting moiety against Clec9A. In another illustrative embodiment, the chimeric protein or chimeric protein complex may include a targeting moiety against CD8 and a targeting moiety against CD3. In another illustrative embodiment, the chimeric protein or chimeric protein complex may include a targeting moiety against CD8 and a targeting moiety against PD-1.

[0296] In various embodiments, the chimeric proteins or the chimeric protein complexes disclosed herein include a CD8 binding agent that is a protein-based agent capable of specific binding to CD8. In various embodiments, the present CDS binding agent is a protein-based agent capable of specific binding to CD8 without functionally modulating (e.g partial or complete neutralization) CD8.

[0297] In various embodiments, the present CD8 binding agent comprises a targeting moiety capable of specific binding. In various embodiments, the CD8 binding agent comprises a targeting moiety having an antigen recognition domain such as an antibody or derivatives thereof. In an embodiment, the CD8 binding agent comprises a targeting moiety, which is an antibody. In various embodiments, the antibody is a full-length multimeric protein that includes two heavy chains and two light chains. Each heavy chain includes one variable region (e.g., VH) and at least three constant regions (e.g., CH 1 , CH2and CH3), and each light chain includes one variable region (VL) and one constant region (CL). The variable regions determine the specificity of the antibody. Each variable region comprises three hypervariable regions also known as complementarity-determining regions (CDRs) flanked by four relatively conserved framework regions (FRs). The three CDRs, referred to as CDR1, CDR2, and CDR3, contribute to the antibody binding specificity. In some embodiments, the antibody is a chimeric antibody In some embodiments, the antibody is a humanized antibody.

[0298] In some embodiments, the CD8 binding agent comprises a targeting moiety, which is an antibody derivative or format. In some embodiments, the present CD8 binding agent comprises a targeting moiety which is a singledomain antibody, a recombinant heavy-chain-only antibody (VHH), a single-chain antibody (scFv), a shark heavy- chain-only antibody (VNAR), a microprotein (cysteine knot protein, knottin), a DARPin; a Tetranectin; an Affibody; a Transbody; an Anticalin; an AdNectin; an Affilin; an Affimer, a Microbody; an aptamer; an alterase; a plastic antibody; a phylomer; a stradobody; a maxibody; an evibody; a fynomer, an armadillo repeat protein, a Kunitz domain, an avimer, an atrimer, a probody, an immunobody, a triomab, a troybody; a pepbody; a vaccibody, a UniBody; a DuoBody, a Fv, a Fab, a Fab', a F(ab')2, a peptide mimetic molecule, or a synthetic molecule, as described in US Patent Nos. or Patent Publication Nos. US 7,417,130, US 2004 / 132094, US 5,831 ,012, US 2004 / 023334, US 7,250,297, US 6,818,418, US 2004 / 209243, US 7,838,629, US 7,186,524, US 6,004,746, US 5,475,096, US 2004 / 146938, US 2004 / 157209, US 6,994,982, US 6,794,144, US 2010 / 239633, US 7,803,907, US 2010 / 119446, and / or US 7,166,697, the contents of which are hereby incorporated by reference in their entireties. See also, Storz MAbs. 2011 May-Jun; 3(3): 310-317.

[0299] In some embodiments, the CD8 binding agent comprises a targeting moiety, which is a single-domain antibody, such as a VHH. The VHH may be derived from, for example, an organism that produces VHH antibody such as a camelid, a shark, or the VHH may be a designed VHH VHHs are antibody-derived therapeutic proteins that contain

[0300] DB1 / 162518159.2 42 ORN-109PC / 114384-5109 the unique structural and functional properties of naturally-occurring heavy-chain antibodies. VHH technology is based on fully functional antibodies from camelids that lack light chains. These heavy-chain antibodies contain a single variable domain (VHH) and two constant domains (CH2 and CH3).

[0301] In an embodiment, the CD8 binding agent comprises a VHH. In some embodiments, the VHH is a humanized VHH or camelized VHH.

[0302] In some embodiments, the VHH comprises a fully human VH domain, e.g. a HUMABODY (Crescendo Biologies, Cambridge, UK). In some embodiments, fully human VH domain, e.g. a HUMABODY is monovalent, bivalent, or trivalent. In some embodiments, the fully human VH domain, e.g. a HUMABODY is mono- or multi-specific such as monospecific, bispecific, or trispecific. Illustrative fully human VH domains, e.g. a HUMABODIES are described in, for example, WO2016 / 113555 and WO2016 / 113557, the entire disclosure of which is incorporated by reference.

[0303] In some embodiments, the CD8 binding agent comprises a targeting moiety which is a VHH comprising a single amino acid chain having four “framework regions” or FRs and three “complementary determining regions” or CDRs. As used herein, “framework region” or “FR” refers to a region in the variable domain, which is located between the CDRs. As used herein, “complementary determining region” or “CDR” refers to variable regions in VHHs that contains the amino acid sequences capable of specifically binding to antigenic targets. In various embodiments, the CD8 binding agent comprises a VHH having a variable domain comprising at least one CDR1 , CDR2, and / or CDR3 sequences.

[0304] In some embodiments, the targeting moiety comprises anti-CD8 antibody as described in WO 2019033043, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD8 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDR H 1 : GFNI KDTYIH (SEQ ID NO: 293); CDR H2: RIDPANDNTLYASKFQG (SEO ID NO: 294); CDR H2: RIDPANDNTLYARKFQG (SEQ ID NO: 295); CDR H3: GRGYGYYVFDH (SEQ ID NO: 296); or CDR H3: TRGYGYYVFDT (SEQ ID NO: 297).

[0305] In some embodiments, the anti-CD8 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDR L1 : SISQY (SEQ ID NO: 298); CDR L1 : SISKY (SEQ ID NO: 299); CDR L2: SGSTLQ (SEQ ID NO: 300); CDR L3: HNENPL (SEQ ID NO: 301); CDR L3: HNEFPV (SEQ ID NO: 302); CDR L3: HNEFPP (SEQ ID NO: 303); CDR L3: VNEFPP (SEQ ID NO: 304); CDR L3: VNEFPV (SEQ ID NO: 305).

[0306] In some embodiments, the targeting moiety comprises anti-CD8 antibody as described in WO2019023148, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD8 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDR H1 : GFIFSNYG (SEQ ID NO: 306); CDR H2: IWYDGSNK (SEQ ID NO: 307); CDR H3: ARSYDMLTGSGDYYGL (SEQ ID NO: 308). In some embodiments, the anti-CD8 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDR L1 : QDITNY (SEQ ID NO: 309); CDR L2: GAS; CDR L3: QQYNNYPLT (SEQ ID NO: 310).

[0307] DB1 / 162518159.2 43 ORN-109PC / 114384-5109

[0308] In some embodiments, the targeting moiety comprises anti-CD8 antibody as described in WO2015184203, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD8 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDR H1 : SGYTGTDYNMH (SEQ ID NO: 311); CDR H2: YIYPYTGGTGYNQKFKN (SEQ ID NO: 312); CDR H1 : DFGMN (SEQ ID NO: 313); CDR H2: LIYYDGSNKFY (SEQ ID NO: 314); CDR H3: PHYDGYYHFFDS (SEQ ID NO: 315). In some embodiments, the anti-CD8 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDR L1 : RASESVDSYDNSLMH (SEQ ID NO: 316); CDR L2: LASNLES (SEQ ID NO: 317); CDR L3: QQNNEDPYT (SEQ ID NO: 318); CDR L1 : KGSQDINNYLA (SEQ ID NO: 319); CDR L2: NTDILHT (SEQ ID NO: 320); CDR L3: YQYNNGYT (SEQ ID NO: 321).

[0309] In some embodiments, the targeting moiety comprises anti-CD8 antibody as described in WO2018170096, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD8 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDR H1 : GYTFTSY (SEQ ID NO: 322); CDR H2: DPSDNY (SEQ ID NO: 333); CDR H3: PKSAYAFDVGGYAMDY (SEQ ID NO: 334). In some embodiments, the anti-CD8 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDR L1: RTSENIDSYLT (SEQ ID NO: 335); CDR L2: AATLLAD (SEQ ID NO: 336); CDR L3: QHYYSTPWT (SEQ ID NO: 337).

[0310] In some embodiments, the targeting moiety comprises anti-CD8 antibody as described in WO2014164553, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD8 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDR H1 : GFNIKD (SEQ ID NO: 338); CDR H2: RIDPANDNT (SEQ ID NO: 339); CDR H3: GYGYYVFDH (SEQ ID NO: 340). In some embodiments, the anti-CD8 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDR L1 : RTSRSISQYLA (SEQ ID NO: 341); CDR L2: SGSTLQS (SEQ ID NO: 342); CDR L3: QQHNENPLT (SEQ ID NO: 343).

[0311] In various embodiments, the chimeric proteins or the chimeric protein complexes disclosed herein include a CD4 binding agent that is a protein-based agent capable of specific binding to CD4. In various embodiments, the present CD4 binding agent is a protein-based agent capable of specific binding to CD4 without functionally modulating (e.g partial or complete neutralization) CD4.

[0312] In various embodiments, the present CD4 binding agent comprises a targeting moiety capable of specific binding. In various embodiments, the CD4 binding agent comprises a targeting moiety having an antigen recognition domain such as an antibody or derivatives thereof. In an embodiment, the CD4 binding agent comprises a targeting moiety, which is an antibody. In various embodiments, the antibody is a full-length multimeric protein that includes two heavy chains and two light chains. Each heavy chain includes one variable region (e.g., VH) and at least three constant regions (e.g., CH 1 , CH2and CH3), and each light chain includes one variable region (VL) and one constant region (CL). The variable regions determine the specificity of the antibody. Each variable region comprises three hypervariable regions also known as complementarity-determining regions (CDRs) flanked by four relatively

[0313] DB1 / 162518159.2 44 ORN-109PC / 114384-5109 conserved framework regions (FRs). The three CDRs, referred to as CDR1, CDR2, and CDR3, contribute to the antibody binding specificity. In some embodiments, the antibody is a chimeric antibody In some embodiments, the antibody is a humanized antibody.

[0314] In some embodiments, the CD4 binding agent comprises a targeting moiety, which is an antibody derivative or format. In some embodiments, the present CD4 binding agent comprises a targeting moiety which is a singledomain antibody, a recombinant heavy-chain-only antibody (VHH), a single-chain antibody (scFv), a shark heavy- chain-only antibody (VNAR), a microprotein (cysteine knot protein, knottin), a DARPin; a Tetranectin; an Affibody; a Transbody; an Anticalin; an AdNectin; an Affilin; an Affimer, a Microbody; an aptamer; an alterase; a plastic antibody; a phylomer; a stradobody; a maxibody; an evibody; a fynomer, an armadillo repeat protein, a Kunitz domain, an avimer, an atrimer, a probody, an immunobody, a triomab, a troybody; a pepbody; a vaccibody, a UniBody; a DuoBody, a Fv, a Fab, a Fab1, a F(ab')2, a peptide mimetic molecule, or a synthetic molecule, as described in US Patent Nos. or Patent Publication Nos. US 7,417,130, US 2004 / 132094, US 5,831 ,012, US 2004 / 023334, US 7,250,297, US 6,818,418, US 2004 / 209243, US 7,838,629, US 7,186,524, US 6,004,746, US 5,475,096, US 2004 / 146938, US 2004 / 157209, US 6,994,982, US 6,794,144, US 2010 / 239633, US 7,803,907, US 2010 / 119446, and / or US 7,166,697, the contents of which are hereby incorporated by reference in their entireties. See also, Storz MAbs. 2011 May-Jun; 3(3): 310-317.

[0315] In some embodiments, the CD4 binding agent comprises a targeting moiety, which is a single-domain antibody, such as a VHH. The VHH may be derived from, for example, an organism that produces VHH antibody such as a camelid, a shark, or the VHH may be a designed VHH VHHs are antibody-derived therapeutic proteins that contain the unique structural and functional properties of naturally-occurring heavy-chain antibodies. VHH technology is based on fully functional antibodies from camelids that lack light chains. These heavy-chain antibodies contain a single variable domain (VHH) and two constant domains (CH2 and CH3).

[0316] In an embodiment, the CD4 binding agent comprises a VHH. In some embodiments, the VHH is a humanized VHH or camelized VHH. In some embodiments, the VHH comprises a fully human V domain, e.g. a HUMABODY (Crescendo Biologies, Cambridge, UK). In some embodiments, fully human V domain, e.g. a HUMABODY is monovalent, bivalent, or trivalent. In some embodiments, the fully human V domain, e.g. a HUMABODY is mono- or multi-specific such as monospecific, bispecific, or trispecific. Illustrative fully human VHdomains, e.g. a HUMABODIES are described in, for example, WO2016 / 113555 and WO2016 / 113557, the entire disclosure of which is incorporated by reference.

[0317] In some embodiments, the CD4 binding agent comprises a targeting moiety which is a VHH comprising a single amino acid chain having four “framework regions” or FRs and three “complementary determining regions” or CDRs. As used herein, “framework region" or “FR” refers to a region in the variable domain, which is located between the CDRs. As used herein, “complementary determining region” or “CDR” refers to variable regions in VHHs that contains the amino acid sequences capable of specifically binding to antigenic targets. In various embodiments, the CD4 binding agent comprises a VHH having a variable domain comprising at least one CDR1, CDR2, and / or

[0318] DB1 / 162518159.2 45 ORN-109PC / 114384-5109

[0319] CDR3 sequences. In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in W02020082045, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDRH1: GYTFTAHI (SEQ ID NO: 344); CDRH2: IKPQYGAV (SEQ ID NO: 345); or CDRH3: AR. In some embodiments, the anti-CD4 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDRL1: QGVGSD (SEQ ID NO: 346); CDRL2: HTS; or CDRL3: QVLQF (SEQ ID NO: 347).

[0320] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in WO2018170096, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDRH1 : GYTFTSN (SEQ ID NO: 348); CDRH2: YPRSGN (SEQ ID NO: 349); or CDRH3: RVPYFDH (SEQ ID NO: 350). In some embodiments, the anti-CD4 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDRL1 : KASQSVGNNVA (SEQ ID NO: 351); CDRL2: YASNRYT (SEQ ID NO: 352); or CDRL3: QQHYSSPFT (SEQ ID NO: 353).

[0321] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in WO2016156570, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one CDR1 comprising the amino acid sequence of: GYWMY (SEQ ID NO: 354); CDR1 : SYSMG (SEQ ID NO: 355); CDR1 : FNAMG (SEQ ID NO: 356); or CDR1 : VMG. In some embodiments, the anti-CD4 antibody comprises at least one CDR2 comprising the amino acid sequence of CDR2: AISPGGGSTYYPDSVK (SEQ ID NO: 357); CDR2: AISWSGDETSYADSVK (SEQ ID NO: 358); CDR2: TIARAGATKYADSVKG (SEQ ID NO: 359); or CDR2: AVRWSSTGIYYTQYAD (SEQ ID NO: 360). In some embodiments, the anti-CD4 antibody comprises at least one CDR3 comprising the amino acid sequence of CDR3: SLTATHTYEYDY (SEQ ID NO: 361); CDR3: DRWWRPAGLQWDY (SEQ ID NO: 362); CDR3: RVFDLPNDY (SEQ ID NO: 363); or CDR3: DTYNSNPARWDGYDF (SEQ ID NO: 364).

[0322] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in WO2012145238, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDRH1 : AYVIS (SEQ ID NO: 365); CDRH2: EIYPGSGSSYYNEKFKG (SEQ ID NO: 366); or CDRH3: SGDGSKFVY (SEQ ID NO: 367). In some embodiments, the anti-CD4 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDRL1 : KASQSVDYCGDSYMN (SEQ ID NO: 368); CDRL2: VASNLES (SEQ ID NO: 369); or CDRL3: QQSLQDPPT (SEQ ID NO: 370).

[0323] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in W02008134046, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDRH1 : GYTFTSYVIH (SEQ ID NO: 371); CDRH2: YINPYNDGTDYDEKFK (SEQ ID NO: 372); or CDRH3: EKDNYATGAWFAY (SEQ ID NO: 373). In some embodiments, the anti-CD4 antibody comprises at least one light chain variable region

[0324] DB1 / 162518159.2 46 ORN-109PC / 114384-5109 comprising the amino acid sequence of CDRL1 : KSSQSLLYSTNQKNY (SEQ ID NO: 374); CDRL2: WASTRES (SEQ ID NO: 375); or CDRL3: QQYYSYRT (SEQ ID NO: 376).

[0325] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in W02009012944, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDRH1 : SYVIH (SEQ ID NO: 377); CDRH1: GFTFSNYAMS (SEQ ID NO: 378); or CDRH2: AISDHSTNTYYP (SEQ ID NO: 379); CDRH3: EKDNYATGAWFAY (SEQ ID NO: 380); or CDRH3: ARKYGGDYDPF (SEQ ID NO: 381). In some embodiments, the anti-CD4 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDRL1 : KSSQSLLYSTNQKNYL (SEQ ID NO: 382); CDRL1 : KSSGSLLYSTNQKNYL (SEQ ID NO: 383); CDRL1 : KASQDINNY (SEQ ID NO: 384); CDRL2: WASTRES (SEQ ID NO: 385); CDRL2: YTSTLQPGVPS (SEQ ID NO: 386); CDRL3: QQYYSYRT (SEQ ID NO: 387); or CDRL3: YDNLLF (SEQ ID NO: 388).

[0326] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in W02004005350, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDRH1 : TFGVH (SEQ ID NO: 389); CDRH1 : TAGVH (SEQ ID NO: 390); or CDRH1 : TFGVA (SEQ ID NO: 391); CDRH2: VIWRSGITDYNVPFMS (SEQ ID NO: 392); CDRH2: VIARSGITDYNVPFMS (SEQ ID NO: 393); CDRH2: VIWASGITDYNVPFMS (SEQ ID NO: 394); CDRH3: NDPGTGFAY (SEQ ID NO: 395); CDRH3: NDPGTGAAY (SEQ ID NO: 396); or CDRH3: NDPGTGFAA (SEQ ID NO: 397). In some embodiments, the anti-CD4 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDRL1 : RASENIYSYLA (SEQ ID NO: 398); CDRL1 : RASENIYSALA (SEQ ID NO: 399); CDRL2: DAKTLAE (SEQ ID NO: 400); CDRL3: QHHYGNPPT (SEQ ID NO: 401); CDRL3: QHAYGNPPT (SEQ ID NO: 402); or CDRL3: QHHAGNPPT (SEQ ID NO: 403).

[0327] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in W02004083247, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain variable region comprising the amino acid sequence of CDRH1 : DYVIN (SEQ ID NO: 404); CDRH2: EIYPGSGSDYYNENLKD (SEQ ID NO: 405); or CDRH3: KGENGNSLAFAY (SEQ ID NO: 406). In some embodiments, the anti-CD4 antibody comprises at least one light chain variable region comprising the amino acid sequence of CDRL1 : QSVDYDGDSYMN (SEQ ID NO: 407); CDRL2: AASNLES (SEQ ID NO: 408); or CDRL3: QQSIQDPCT (SEQ ID NO: 409).

[0328] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in W02014100139, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain comprising the following amino acid sequence:

[0329] Anti-CD4 antibody MV1, Heavy Chain

[0330] MEWSGVFMFLLSVTAGVHSQVQLQQSGPEWKPGASVKMSCKASGYTFTSYVIHWVRQKP

[0331] GQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNY

[0332] DB1 / 162518159.2 47 ORN-109PC / 114384-5109

[0333] ATGAWFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREP QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 410)

[0334] In some embodiments, the anti-CD4 antibody comprises at least one light chain comprising the following amino acid sequence:

[0335] Anti-CD4 antibody MV1, Light Chain

[0336] MEWSGVFIFLLSVTAGVHSDIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQ QKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYYRTFGGG TKLEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 411)

[0337] In some embodiments, the targeting moiety comprises anti-CD4 antibody as described in WQ2004083247, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the anti-CD4 antibody comprises at least one heavy chain comprising the following amino acid sequence:

[0338] EEQLVESGGGLVKPGGSLRLSCAASGFSFSDCRMYWLRQAPGKGLEWIGVISVKSENYGAN YAESVRGRFTISRDDSKNTVYLQMNSLKTEDTAVYYCSASYYRYDVGAFAYGQGTLVTVSS (SEQ ID NO: 412)

[0339] In some embodiments, the anti-CD4 antibody comprises at least one light chain comprising the following amino acid sequence:

[0340] DIVMTQSPDSLAVSLGERATINCRASKSVSTSGYSYIYWYQQKPGQPPKLLIYLASILESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQHSRELPTFGQGTKVEIK (SEQ ID NO: 413)

[0341] In one embodiment, the chimeric proteins or chimeric protein complexes comprises one or more targeting moieties directed against a B cell and one or more targeting moieties directed against the same or another B cell. In one embodiment, the chimeric proteins or chimeric protein complexes comprises one or more targeting moieties directed against a B cell and one or more targeting moieties directed against a T cell. In one embodiment, the chimeric proteins or chimeric protein complexes comprises one or more targeting moieties directed against a B cell and one or more targeting moieties directed against a dendritic cell. In one embodiment, the chimeric proteins or chimeric protein complexes comprises one or more targeting moieties against a B cell and one or more targeting moieties directed against a macrophage. In one embodiment, the chimeric proteins or chimeric protein complexes comprises one or more targeting moieties against a B cell and one or more targeting moieties directed against a NK cell.

[0342] DB1 / 162518159.2 48 ORN-109PC / 114384-5109

[0343] In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a dendritic cell and one or more targeting moieties directed against the same or another dendritic cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a dendritic cell and one or more targeting moieties directed against a T cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a dendritic cell and one or more targeting moieties directed against a B cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against a dendritic cell and one or more targeting moieties directed against a macrophage In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against a dendritic cell and one or more targeting moieties directed against a NK cell.

[0344] In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a macrophage and one or more targeting moieties directed against the same or another macrophage. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a macrophage and one or more targeting moieties directed against a T cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against a macrophage and one or more targeting moieties directed against a B cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against a macrophage and one or more targeting moieties directed against a dendritic cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against a macrophage and one or more targeting moieties directed against a NK cell.

[0345] In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against an NK cell and one or more targeting moieties directed against the same or another NK cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against an NK cell and one or more targeting moieties directed against a T cell. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties directed against an NK cell and one or more targeting moieties directed against a B cell In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against an NK cell and one or more targeting moieties directed against a macrophage. In one embodiment, the chimeric protein or chimeric protein complex comprises one or more targeting moieties against an NK cell and one or more targeting moieties directed against a dendritic cell.

[0346] In one embodiment, the chimeric protein or chimeric protein complex comprises a targeting moiety directed against a tumor cell and a second targeting moiety directed against the same or a different tumor cell. In such embodiments, the targeting moieties may bind to any of the tumor antigens described herein.

[0347] In some embodiments, the chimeric protein or chimeric protein complex of the invention comprises one or more targeting moieties having recognition domains that bind to a target (e.g. antigen, receptor) of interest including

[0348] DB1 / 162518159.2 49 ORN-109PC / 114384-5109 those found on one or more cells selected from adipocytes (e.g., white fat cell, brown fat cell), liver lipocytes, hepatic cells, kidney cells (e.g., kidney parietal cell, kidney salivary gland, mammary gland, etc.), duct cells (of seminal vesicle, prostate gland, etc.), intestinal brush border cells (with microvilli), exocrine gland striated duct cells, gall bladder epithelial cells, ductulus efferens nonciliated cells, epididymal principal cells, epididymal basal cells, endothelial cells, ameloblast epithelial cells (tooth enamel secretion), planum semilunatum epithelial cells of vestibular system of ear (proteoglycan secretion), organ of Corti interdental epithelial cells (secreting tectorial membrane covering hair cells), loose connective tissue fibroblasts, corneal fibroblasts (corneal keratocytes), tendon fibroblasts, bone marrow reticular tissue fibroblasts, nonepithelial fibroblasts, pericytes, nucleus pulposus cells of intervertebral disc, cementoblasts / cementocytes (tooth root bonelike ewan cell secretion), odontoblasts / odontocytes (tooth dentin secretion), hyaline cartilage chondrocytes, fi brocartilage chondrocytes, elastic cartilage chondrocytes, osteoblasts / osteocytes, osteoprogenitor cells (stem cell of osteoblasts), hyalocytes of vitreous body of eye, stellate cells of perilymphatic space of ear, hepatic stellate cells (Ito cell), pancreatic stelle cells, skeletal muscle cells, satellite cells, heart muscle cells, smooth muscle cells, myoepithelial cells of iris, myoepithelial cells of exocrine glands, exocrine secretory epithelial cells (e.g. , salivary gland cells, mammary gland cells, lacrimal gland cells, sweat gland cells, sebaceious gland cells, prostate gland cells, gastric glad cells, pancreatic acinar cells, pneumocytes), a hormone secreting cells (e.g., pituitary cells, neurosecretory cells, gut and respiratory tract cells, thyroid gland cells, parathyroid glad cells, adrenal gland cells, Leydig cells of testes, pancreatic islet cells), keratinizing epithelial cells, wet stratified barrier epithelial cells, neuronal cells (e.g., sensory transducer cells, autonomic neuron cells, sense organ and peripheral neuron supporting cells, and central nervous system neurons and glial cells such as interneurons, principal cells, astrocytes, oligodendrocytes, and ependymal cells).

[0349] Targeting Moiety Formats

[0350] In various embodiments, the targeting moiety of the chimeric protein or chimeric protein complex is a protein-based agent capable of specific binding, such as an antibody or derivatives thereof. In an embodiment, the targeting moiety comprises an antibody In various embodiments, the antibody is a full-length multimeric protein that includes two heavy chains and two light chains. Each heavy chain includes one variable region (e.g., VH) and at least three constant regions (e.g., CH 1 , CH2 and CH3), and each light chain includes one variable region (VL) and one constant region (CL) The variable regions determine the specificity of the antibody. Each variable region comprises three hypervariable regions also known as complementarity-determining regions (CDRs) flanked by four relatively conserved framework regions (FRs). The three CDRs, referred to as CDR1 , CDR2, and CDR3, contribute to the antibody binding specificity. In some embodiments, the antibody is a chimeric antibody In some embodiments, the antibody is a humanized antibody.

[0351] In some embodiments, the targeting moiety comprises antibody derivatives or formats. In some embodiments, the targeting moiety of the chimeric protein or chimeric protein complex is a single-domain antibody, a recombinant heavy-chain-only antibody (VHH), a single-chain antibody (scFv), a shark heavy-chain-only antibody (VNAR), a microprotein (cysteine knot protein, knottin), a DARPin; a Tetranectin; an Affibody; a Transbody; an Anticalin; an

[0352] DB1 / 162518159.2 50 ORN-109PC / 114384-5109

[0353] AdNectin; an Affilin; a Microbody; a peptide aptamer; an alterases; a plastic antibodies; a phylomer; a stradobodies; a maxibodies; an evibody; a fynomer, an armadillo repeat protein, a Kunitz domain, an avimer, an atrimer, a probody, an immunobody, a triomab, a troybody; a pepbody; a vaccibody, a UniBody; affimers, a DuoBody, a Fv, a Fab, a Fab', a F(ab')2, a peptide mimetic molecule, or a synthetic molecule, as described in US Patent Nos. or Patent Publication Nos. US 7,417,130, US 2004 / 132094, US 5,831 ,012, US 2004 / 023334, US 7,250,297, US 6,818,418, US 2004 / 209243, US 7,838,629, US 7,186,524, US 6,004,746, US 5,475,096, US 2004 / 146938, US 2004 / 157209, US 6,994,982, US 6,794,144, US 2010 / 239633, US 7,803,907, US 2010 / 119446, and / or US 7,166,697, the contents of which are hereby incorporated by reference in their entireties. See also, Storz MAbs. 2011 May-Jun; 3(3): 310-317.

[0354] In one embodiment, the targeting moiety comprises a single-domain antibody, such as VHH from, for example, an organism that produces VHH antibody such as a camelid, a shark, or a designed VHH. VHHs are antibody-derived therapeutic proteins that contain the unique structural and functional properties of naturally-occurring heavy-chain antibodies. VHH technology is based on fully functional antibodies from camelids that lack light chains. These heavy-chain antibodies contain a single variable domain (VHH) and two constant domains (CH2 and CH3). VHHs are commercially available under the trademark of NANOBODY or NANOBODIES.

[0355] In an embodiment, the targeting moiety comprises a VHH. In some embodiments, the VHH is a humanized VHH or camelized VHH.

[0356] In some embodiments, the VHH comprises a fully human VH domain, e.g. a HUMABODY (Crescendo Biologies, Cambridge, UK). In some embodiments, fully human VH domain, e.g. a HUMABODY is monovalent, bivalent, or trivalent. In some embodiments, the fully human VH domain, e.g. a HUMABODY is mono- or multi-specific such as monospecific, bispecific, or trispecific. Illustrative fully human VH domains, e.g. a HUMABODIES are described in, for example, WO 2016 / 113555 and WO2016 / 113557, the entire disclosure of which is incorporated by reference.

[0357] In various embodiments, the targeting moiety of the chimeric protein or chimeric protein complex is a protein-based agent capable of specific binding to a cell receptor, such as a natural ligand for the cell receptor. In various embodiments, the cell receptor is found on one or more immune cells, which can include, without limitation, T cells, cytotoxic T lymphocytes, T helper cells, natural killer (NK) cells, natural killer T (NKT) cells, anti-tumor macrophages (e.g. M1 macrophages), B cells, dendritic cells, or subsets thereof. In some embodiments, the cell receptor is found on megakaryocytes, thrombocytes, erythrocytes, mast cells, basophils, neutrophils, eosinophils, or subsets thereof.

[0358] In some embodiments, the targeting moiety is a natural ligand such as a chemokine. Illustrative chemokines that may be included in the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex of the invention include, but are not limited to, CCL1 , CCL2, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9, CCL10, CCL11 , CCL12, CCL13, CCL14, CCL15, CCL16, CL17, CCL18, CCL19, CCL20, CCL21, CCL22, CCL23, CCL24, CLL25, CCL26, CCL27, CXCL1 , CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10,

[0359] DB1 / 162518159.2 51 ORN-109PC / 114384-5109

[0360] CXCL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCL17, XCL1, XCL2, CX3CL1, HCC-4, and LDGF- PBP. In an illustrative embodiment, the targeting moiety may be XCL1 , which is a chemokine that recognizes and binds to the dendritic cell receptor XCR1. In another illustrative embodiment, the targeting moiety is CCL1 , which is a chemokine that recognizes and binds to CCR8. In another illustrative embodiment, the targeting moiety is CCL2, which is a chemokine that recognizes and binds to CCR2 or CCR9. In another illustrative embodiment, the targeting moiety is CCL3, which is a chemokine that recognizes and binds to CCR1 , CCR5, or CCR9. In another illustrative embodiment, the targeting moiety is CCL4, which is a chemokine that recognizes and binds to CCR1 or CCR5 or CCR9. In another illustrative embodiment, the targeting moiety is CCL5, which is a chemokine that recognizes and binds to CCR1 or CCR3 or CCR4 or CCR5. In another illustrative embodiment, the targeting moiety is CCL6, which is a chemokine that recognizes and binds to CCR1. In another illustrative embodiment, the targeting moiety is CCL7, which is a chemokine that recognizes and binds to CCR2 or CCR9. In another illustrative embodiment, the targeting moiety is CCL8, which is a chemokine that recognizes and binds to CCR1 or CCR2 or CCR2B or CCR5 or CCR9. In another illustrative embodiment, the targeting moiety is CCL9, which is a chemokine that recognizes and binds to CCR1 . In another illustrative embodiment, the targeting moiety is CCL10, which is a chemokine that recognizes and binds to CCR1. In another illustrative embodiment, the targeting moiety is CCL11 , which is a chemokine that recognizes and binds to CCR2 or CCR3 or CCR5 or CCR9 In another illustrative embodiment, the targeting moiety is CCL13, which is a chemokine that recognizes and binds to CCR2 orCCR3 or CCR5 or CCR9. In another illustrative embodiment, the targeting moiety is CCL14, which is a chemokine that recognizes and binds to CCR1 or CCR9. In another illustrative embodiment, the targeting moiety is CCL15, which is a chemokine that recognizes and binds to CCR1 or CCR3. In another illustrative embodiment, the targeting moiety is CCL16, which is a chemokine that recognizes and binds to CCR1, CCR2, CCR5, or CCR8. In another illustrative embodiment, the targeting moiety is CCL17, which is a chemokine that recognizes and binds to CCR4. In another illustrative embodiment, the targeting moiety is CCL19, which is a chemokine that recognizes and binds to CCR7 In another illustrative embodiment, the targeting moiety is CCL20, which is a chemokine that recognizes and binds to CCR6. In another illustrative embodiment, the targeting moiety is CCL21, which is a chemokine that recognizes and binds to CCR7. In another illustrative embodiment, the targeting moiety is CCL22, which is a chemokine that recognizes and binds to CCR4. In another illustrative embodiment, the targeting moiety is CCL23, which is a chemokine that recognizes and binds to CCR1 . In another illustrative embodiment, the targeting moiety is CCL24, which is a chemokine that recognizes and binds to CCR3. In another illustrative embodiment, the targeting moiety is CCL25, which is a chemokine that recognizes and binds to CCR9. In another illustrative embodiment, the targeting moiety is CCL26, which is a chemokine that recognizes and binds to CCR3. In another illustrative embodiment, the targeting moiety is CCL27, which is a chemokine that recognizes and binds to CCR10. In another illustrative embodiment, the targeting moiety is CCL28, which is a chemokine that recognizes and binds to CCR3 or CCR10. In another illustrative embodiment, the targeting moiety is CXCL1 , which is a chemokine that recognizes and binds to CXCR1 or CXCR2. In another illustrative embodiment, the targeting moiety is CXCL2, which is a chemokine that recognizes and binds to CXCR2. In another illustrative embodiment, the targeting moiety is CXCL3, which is a chemokine that recognizes and binds to CXCR2. In another illustrative embodiment, the

[0361] DB1 / 162518159.2 52 ORN-109PC / 114384-5109 targeting moiety is CXCL4, which is a chemokine that recognizes and binds to CXCR3B. In another illustrative embodiment, the targeting moiety is CXCL5, which is a chemokine that recognizes and binds to CXCR2 In another illustrative embodiment, the targeting moiety is CXCL6, which is a chemokine that recognizes and binds to CXCR1 orCXCR2. In another illustrative embodiment, the targeting moiety is CXCL8, which is a chemokine that recognizes and binds to CXCR1 or CXCR2. In another illustrative embodiment, the targeting moiety is CXCL9, which is a chemokine that recognizes and binds to CXCR3. In another illustrative embodiment, the targeting moiety is CXCL10, which is a chemokine that recognizes and binds to CXCR3. In another illustrative embodiment, the targeting moiety is CXCL11, which is a chemokine that recognizes and binds to CXCR3 or CXCR7. In another illustrative embodiment, the targeting moiety is CXCL12, which is a chemokine that recognizes and binds to CXCR4 or CXCR7 In another illustrative embodiment, the targeting moiety is CXCL13, which is a chemokine that recognizes and binds to CXCR5. In another illustrative embodiment, the targeting moiety is CXCL16, which is a chemokine that recognizes and binds to CXCR6. In another illustrative embodiment, the targeting moiety is LDGF- PBP, which is a chemokine that recognizes and binds to CXCR2. In another illustrative embodiment, the targeting moiety is XCL2, which is a chemokine that recognizes and binds to XCR1 . In another illustrative embodiment, the targeting moiety is CX3CL1, which is a chemokine that recognizes and binds to CX3CR1 .

[0362] In some embodiments, the targeting moiety is a natural ligand such as FMS-like tyrosine kinase 3 ligand (Flt3L) or a truncated region thereof (e.g., which is able to bind Flt3). In some embodiments, the targeting moiety is an extracellular domain of Flt3L. In some embodiments, the targeting moiety comprising a Flt3L domain, wherein the Flt3L domain is a single chain dimer, optionally where one Flt3L domain is connoted to the other Flt3L domain via one or more linkers, wherein the linker is a flexible linker. In some embodiments, the targeting moiety of the present invention comprises Flt3L domain, wherein the Flt3L domain is a single chain dimer and an Fc domain, the Fc domain optionally having one or more mutations that reduces or eliminates one or more effector functions of the Fc domain, promotes Fc chain pairing in the Fc domain, and / or stabilizes a hinge region in the Fc domain. In some embodiments, the targeting moiety recognizes CD20. In some embodiments, the targeting moiety recognizes PD- L1 . In some embodiments, the targeting moiety recognizes Clec9A.

[0363] In various embodiments, the chimeric protein or chimeric protein complex comprises targeting moieties in various combinations. In an illustrative embodiment, the chimeric protein or chimeric protein complex may comprise two targeting moieties, wherein both targeting moieties are antibodies or derivatives thereof In another illustrative embodiment, the chimeric protein or chimeric protein complex may comprise two targeting moieties, wherein both targeting moieties are natural ligands for cell receptors. In a further illustrative embodiment, the chimeric protein or chimeric protein complex may comprise two targeting moieties, wherein one of the targeting moieties is an antibody or derivative thereof, and the other targeting moiety is a natural ligand for a cell receptor

[0364] In various embodiments, the recognition domain of the chimeric protein or chimeric protein complex functionally modulates (by way of non-limitation, partially or completely neutralizes) the target (e.g. antigen, receptor) of interest, e.g. substantially inhibiting, reducing, or neutralizing a biological effect that the antigen has. For example, various recognition domains may be directed against one or more tumor antigens that are actively suppressing, or

[0365] DB1 / 162518159.2 53 ORN-109PC / 114384-5109 have the capacity to suppress, the immune system of, for example, a patient bearing a tumor. For example, in some embodiments, the chimeric protein or chimeric protein complex functionally modulates immune inhibitory signals (e.g. checkpoint inhibitors), for example, one or more of TIM-3, BTLA, PD-1, CTLA-4, B7-H4, GITR, galectin-9, HVEM, PD-L1, PD-L2, B7-H3, CD244, CD160, TIGIT, SIRPa, ICOS, CD172a, and TMIGD2. For example, in some embodiments, the chimeric protein or chimeric protein complex is engineered to disrupt, block, reduce, and / or inhibit the transmission of an immune inhibitory signal, by way of non-limiting example, the binding of PD-1 with PD-L1 or PD-L2 and / or the binding of CTLA-4 with one or more of AP2M1, CD80, CD86, SHP-2, and PPP2R5A.

[0366] In various embodiments, the recognition domain of the chimeric protein or chimeric protein complex binds but does not functionally modulate the target (e.g. antigen, receptor) of interest, e.g. the recognition domain is, or is akin to, a binding antibody. For instance, in various embodiments, the recognition domain simply targets the antigen or receptor but does not substantially inhibit, reduce or functionally modulate a biological effect that the antigen or receptor has. For example, some of the smaller antibody formats described above (e.g. as compared to, for example, full antibodies) have the ability to target hard to access epitopes and provide a larger spectrum of specific binding locales. In various embodiments, the recognition domain binds an epitope that is physically separate from an antigen or receptor site that is important for its biological activity (e.g. the antigen’s active site).

[0367] Such non-neutralizing binding finds use in various embodiments of the present invention, including methods in which the chimeric protein or chimeric protein complex is used to directly or indirectly recruit active immune cells to a site of need via an effector antigen, such as any of those described herein. For example, in various embodiments, the chimeric protein or chimeric protein complex such as Fc-based chimeric protein complex may be used to directly or indirectly recruit cytotoxic T cells via CD8 to a tumor cell in a method of reducing or eliminating a tumor (e.g. the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex may comprise an anti-CD8 recognition domain and a recognition domain directed against a tumor antigen) In such embodiments, it is desirable to directly or indirectly recruit CD8-expressing cytotoxic T cells but not to functionally modulate the CD8 activity. On the contrary, in these embodiments, CD8 signaling is an important piece of the tumor reducing or eliminating effect. By way of further example, in various methods of reducing or eliminating tumors, the chimeric protein or chimeric protein complex is used to directly or indirectly recruit dendritic cells (DCs) via CLEC9A (e.g. the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex may comprise an anti-CLEC9A recognition domain and a recognition domain directed against a tumor antigen). In such embodiments, it is desirable to directly or indirectly recruit CLEC9A-expressing DCs but not to functionally modulate the CLEC9A activity. On the contrary, in these embodiments, CLEC9A signaling is an important piece of the tumor reducing or eliminating effect.

[0368] In various embodiments, the recognition domain of the chimeric protein or chimeric protein complex binds to XCR1 e.g. on dendritic cells. For instance, the recognition domain, in some embodiments comprises all or part of XCL1 or a non-neutralizing anti-XCR1 agent.

[0369] DB1 / 162518159.2 54 ORN-109PC / 114384-5109

[0370] In various embodiments, the recognition domain of the chimeric protein or chimeric protein complex binds to an immune modulatory antigen (e.g. immune stimulatory or immune inhibitory). In various embodiments, the immune modulatory antigen is one or more of 4-1 BB, OX-40, HVEM, GITR, CD27, CD28, CD30, CD40, ICOS ligand; OX- 40 ligand, LIGHT (CD258), GITR ligand, CD70, B7-1, B7-2, CD30 ligand, CD40 ligand, ICOS, ICOS ligand, CD137 ligand and TL1A. In various embodiments, such immune stimulatory antigens are expressed on a tumor cell. In various embodiments, the recognition domain of the chimeric protein or chimeric protein complex binds but does not functionally modulate such immune stimulatory antigens and therefore allows recruitment of cells expressing these antigens without the reduction or loss of their potential tumor reducing or eliminating capacity.

[0371] In various embodiments, the recognition domain of the chimeric protein or chimeric protein complex may be in the context of chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex that comprises two recognition domains that have neutralizing activity, or comprises two recognition domains that have non-neutralizing (e.g. binding) activity, or comprises one recognition domain that has neutralizing activity and one recognition domain that has non-neutralizing (e.g. binding) activity.

[0372] Fc Domains

[0373] The fragment crystallizable domain (Fc domain) is the tail region of an antibody that interacts with Fc receptors located on the cell surface of cells that are involved in the immune system, e.g., B lymphocytes, dendritic cells, natural killer cells, macrophages, neutrophils, eosinophils, basophils, and mast cells. In IgG, IgA and IgD antibody isotypes, the Fc domain is composed of two identical protein fragments, derived from the second and third constant domains of the antibody's two heavy chains In IgM and IgE antibody isotypes, the Fc domain contains three heavy chain constant domains (CH domains 2—4) in each polypeptide chain.

[0374] In some embodiments, the Fc-based chimeric protein of complex the present technology includes a Fc domain. In some embodiments, the Fc domains are from selected from IgG, IgA, IgD, IgM or IgE. In some embodiments, the Fc domains are from selected from lgG1 , lgG2, lgG3, or lgG4.

[0375] In some embodiments, the Fc domains are from selected from human IgG, IgA, IgD, IgM or IgE. In some embodiments, the Fc domains are from selected from human IgG 1 , lgG2, lgG3, or lgG4

[0376] In some embodiments, the Fc domains of the Fc-based chimeric protein complex comprise the CH2 and CH3 regions of IgG. In some embodiments, the IgG is human IgG. In some embodiments, the human IgG is selected from lgG1 , lgG2, lgG3, or lgG4.

[0377] In some embodiments, the Fc domains comprise one or more mutations. In some embodiments, the mutation(s) to the Fc domains reduces or eliminates the effector function the Fc domains. In some embodiments, the mutated Fc domain has reduced affinity, activity, or binding to a target receptor. By way of example, in some embodiments, the mutation to the Fc domains reduces or eliminates the binding of the Fc domains to FcyR. In some embodiments, the FcyR is selected from FcyRI; FcyRlla, 131 R / R; FcyRlla, 131 H / H, FcyRllb; and FcyRIII. In some embodiments, the mutation to the Fc domains reduces or eliminated binding to complement proteins, such as, e.g., C1q. In some

[0378] DB1 / 162518159.2 55 ORN-109PC / 114384-5109 embodiments, the mutation to the Fc domains reduces or eliminated binding to both FcyR and complement proteins, such as, e.g., C1q.

[0379] In some embodiments, the Fc domains comprise the LALA mutation to reduce or eliminate the effector function of the Fc domains. By way of example, in some embodiments, the LALA mutation comprises L234A and L235A substitutions in human IgG (e.g., lgG1) (wherein the numbering is based on the commonly used numbering of the CH2 residues for human lgG1 according to EU convention (PNAS, Edelman et al., 1969; 63 (1) 78-85)).

[0380] In some embodiments, the Fc domains of human IgG comprise a mutation to reduce or eliminate the effector function of the Fc domains. By way of example, in some embodiments, the mutations are selected from L234A, L234F, L235A, L235E, L235Q, K322A, K322Q, D265A, P329G, P329A, P331G, and P331S.

[0381] In some embodiments, the Fc domains comprise the FALA mutation to reduce or eliminate the effector function of the Fc domains. By way of example, in some embodiments, the FALA mutation comprises F234A and L235A substitutions in human lgG4.

[0382] In some embodiments, the Fc domains of human I gG4 comprise a mutation at one or more of F234, L235, K322, D265, and P329 to reduce or eliminate the effector function of the Fc domains. By way of example, in some embodiments, the mutations are selected from F234A, L235A, L235E, L235Q, K322A, K322Q, D265A, P329G, and P329A.

[0383] In some embodiments, the mutation(s) to the Fc domain stabilize a hinge region in the Fc domain. By way of example, in some embodiments, the Fc domain comprises a mutation at S228 of human IgG to stabilize a hinge region In some embodiments, the mutation is S228P.

[0384] In some embodiments, the mutation(s) to the Fc domain promote chain pairing in the Fc domain. In some embodiments, chain pairing is promoted by ionic pairing (a / k / a charged pairs, ionic bond, or charged residue pair).

[0385] In some embodiments, the Fc domain comprises a mutation at one more of the following amino acid residues of IgG to promote of ionic pairing: D356, E357, L368, K370, K392, D399, and K409.

[0386] By way of example, in some embodiments, the human IgG Fc domain comprise one of the mutation combinations in Table 1 to promote of ionic pairing.

[0387] DBl / 162518159.2 56 ORN-109PC / 114384-5109

[0388] DB1 / 162518159.2 57 ORN-109PC / 114384-5109

[0389] In some embodiments, chain pairing is promoted by a knob-in-hole mutations. In some embodiments, the Fc domain comprises one or more mutations to allow for a knob-in-hole interaction in the Fc domain. In some embodiments, a first Fc chain is engineered to express the “knob" and a second Fc chain is engineered to express the complementary “hole.” By way of example, in some embodiments, human IgG Fc domain comprises the mutations of Table 2 to allow for a knob-in-hole interaction.

[0390] In some embodiments, the Fc domains in the Fc-based chimeric protein complexes of the present technology comprise any combination of the above-disclosed mutations. By way of example, in some embodiments, the Fc domain comprises mutations that promote ionic pairing and / or a knob-in-hole interaction. By way of example, in some embodiments, the Fc domain comprises mutations that have one or more of the following properties: promote ionic pairing, induce a knob-in-hole interaction, reduce or eliminate the effector function of the Fc domain, and cause Fc stabilization (e.g. at hinge).

[0391] DBl / 162518159.2 58 ORN-109PC / 114384-5109

[0392] By way of example, in some embodiments, a human IgG Fc domains comprise mutations disclosed in Table 3, which promote ionic pairing and / or promote a knob-in-hole interaction in the Fc domain.

[0393] DBl / 162518159.2 59 ORN-109PC / 114384-5109

[0394] DB1 / 162518159.2 60 ORN-109PC / 114384-5109

[0395] By way of example, in some embodiments, a human IgG Fc domains comprise mutations disclosed in Table 4, which promote ionic pairing, promote a knob-in-hole interaction, or a combination thereof in the Fc domain. In embodiments, the “Chain 1” and "Chain 2” of Table 4 can be interchanged (e.g. Chain 1 can have Y407T and Chain 2 can have T366Y).

[0396] DBl / 162518159.2 61 ORN-109PC / 114384-5109

[0397] DB1 / 162518159.2 62 ORN-109PC / 114384-5109

[0398] DB1 / 162518159.2 63 ORN-109PC / 114384-5109

[0399] By way of example, in some embodiments, a human IgG Fc domains comprise mutations disclosed in Table 5, which reduce or eliminate FcyR and / or complement binding in the Fc domain. In embodiments, the Table 5 mutations are in both chains.

[0400] DBl / 162518159.2 64 ORN-109PC / 114384-5109

[0401] DB1 / 162518159.2 65 ORN-109PC / 114384-5109

[0402] In some embodiments, the Fc domains in the Fc-based chimeric protein complexes of the present technology are homodimeric, i.e., the Fc region in the chimeric protein complex comprises two identical protein fragments.

[0403] In some embodiments, the Fc domains in the Fc-based chimeric protein complexes of the present technology are heterodimeric, i.e., the Fc domain comprises two non-identical protein fragments.

[0404] In some embodiments, heterodimeric Fc domains are engineered using ionic pairing and / or knob-in-hole mutations described herein. In some embodiments, the heterodimeric Fc-based chimeric protein complexes have a trans orientation / configuration. In a trans orientation / configuration, the targeting moiety and signaling agent, in embodiments, not found on the same polypeptide chain in the present Fc-based chimeric protein complexes. In some embodiments, the Fc domains includes or starts with the core hinge region of wild-type human lgG1 , which contains the sequence Cys-Pro-Pro-Cys. In some embodiments, the Fc domains also include the upper hinge, or parts thereof [e.g., DKTHTCPPC; see WO 2009053368), EPKSCDKTHTCPPC, or EPKSSDKTHTCPPC; see Lo et al., Protein Engineering vol.11 no.6 pp.495-500, 1998)).

[0405] Fc-based Chimeric Protein Complexes

[0406] DBl / 162518159.2 66 ORN-109PC / 114384-5109

[0407] The Fc-based chimeric protein complexes of the present technology comprise at least one Fc domain disclosed herein, at least one antagonistic signaling agent, e.g. IL-6 antagonist, disclosed herein and at least one targeting moiety (TM) disclosed herein.

[0408] It is understood that, the present Fc-based chimeric protein complexes may encompass a complex of two fusion proteins, each comprising an Fc domain. In some embodiments, the Fc-based chimeric protein complex is heterodimeric. In some embodiments, the heterodimeric Fc-based chimeric protein complex has a trans orientation / configuration. In some embodiments, the heterodimeric Fc-based chimeric protein complex has a cis orientation / configuration.

[0409] In some embodiments, heterodimeric Fc domains are engineered using ionic pairing and / or knob-in-hole mutations described herein. In some embodiments, the heterodimeric Fc-based chimeric protein complexes have a trans orientation. In a trans orientation, the targeting moiety and signaling agent are, in embodiments, not found on the same polypeptide chain in the present Fc-based chimeric protein complexes. In a trans orientation, the targeting moiety and signaling agent are, in embodiments, found on separate polypeptide chains in the Fc-based chimeric protein complexes. In a cis orientation, the targeting moiety and signaling agent are, in embodiments, found on the same polypeptide chain in the Fc-based chimeric protein complexes.

[0410] In some embodiments, where more than one targeting moiety is present in the heterodimeric protein complexes described herein, one targeting moiety may be in trans orientation (relative to the signaling agent), whereas another targeting moiety may be in cis orientation (relative to the signaling agent). In some embodiments, the signaling agent and target moiety are on the same ends / sides (N-terminal or C-terminal ends) of an Fc domain. In some embodiments, the signaling agent and targeting moiety are on different sides / ends of a Fc domain (N-terminal and C-terminal ends).

[0411] In some embodiments, where more than one targeting moiety is present in the heterodimeric protein complexes described herein, the targeting moieties may be found on the same Fc chain or on two different Fc chains in the heterodimeric protein complex (in the latter case the targeting moieties would be in trans relative to each other, as they are on different Fc chains). In some embodiments, where more than one targeting moiety is present on the same Fc chain, the targeting moieties may be on the same or different sides / ends of a Fc chain (N-terminal or / and C-terminal ends).

[0412] In some embodiments, where more than one signaling agent is present in the heterodimeric protein complexes described herein, the signaling agents may be found on the same Fc chain or on two different Fc chains in the heterodimeric protein complex (in the latter case the signaling agents would be in trans relative to each other, as they are on different Fc chains) In some embodiments, where more than one signaling agent is present on the same Fc chain, the signaling agents may be on the same or different sides / ends of a Fc chain (N-terminal or / and C-terminal ends).

[0413] DB1 / 162518159.2 67 ORN-109PC / 114384-5109

[0414] In some embodiments, where more than one signaling agent is present in the heterodimeric protein complexes described herein, one signaling agent may be in trans orientation (as relates to the targeting moiety), whereas another signaling agent may be in cis orientation (as relates to the targeting moiety).

[0415] In some embodiments, the heterodimeric Fc-based chimeric protein complex does not comprise the antagonistic signaling agent, e.g. IL-6 antagonist, and targeting moiety on a single polypeptide.

[0416] In some embodiments, the Fc-based chimeric protein has an improved in vivo half-life relative to a chimeric protein lacking an Fc or a chimeric protein, which is not a heterodimeric complex. In some embodiments, the Fc-based chimeric protein has an improved solubility, stability and other pharmacological properties relative to a chimeric protein lacking an Fc or a chimeric protein, which is not a heterodimeric complex.

[0417] Heterodimeric Fc-based chimeric protein complexes are composed of two different polypeptides. In embodiments described herein, the targeting domain is on a different polypeptide than the antagonistic signaling agent, e.g. IL- 6 antagonist, and accordingly, proteins that contain only one targeting domain copy, and also only one antagonistic signaling agent, e.g. IL-6 antagonist, copy can be made (this provides a configuration in which potential interference with desired properties can be controlled). Further, in embodiments, one targeting domain (e.g. VHH) only can avoid cross-linking of the antigen on the cell surface (which could elicit undesired effects in some cases). Further, in embodiments, one antagonistic signaling agent, e.g. IL-6 antagonist, may alleviate molecular “crowding” and potential interference with avidity mediated induction or restoration of effector function in dependence of the targeting domain. Further, in embodiments, heterodimeric Fc-based chimeric protein complexes can have two targeting moieties and these can be placed on the two different polypeptides. For instance, in embodiments, the C-terminus of both targeting moieties (e.g. VHHs) can be masked to avoid potential autoantibodies or pre-existing antibodies (e.g. VHH autoantibodies or pre-existing antibodies). Further, in embodiments, heterodimeric Fc-based chimeric protein complexes, e.g. with the targeting domain on a different polypeptide than the antagonistic signaling agent, e.g. IL-6 antagonist, may favor “cross-linking” of two cell types (e.g. a tumor cell and an immune cell). Further, in embodiments, heterodimeric Fc-based chimeric protein complexes can have two signaling agent, each on different polypeptides to allow more complex effector responses.

[0418] Further, in embodiments, heterodimeric Fc-based chimeric protein complexes, e.g. with the targeting domain on a different polypeptide than the antagonistic signaling agent, e.g. IL-6 antagonist, combinatorial diversity of targeting moiety and antagonistic signaling agent, e.g. IL-6 antagonist, is provided in a practical manner. For instance, in embodiments, polypeptides with any of the targeting moieties described herein can be combined “off the shelf” with polypeptides with any of the signaling agents described herein to allow rapid generation of various combinations of targeting moieties and signaling agents in single Fc-based chimeric protein complexes.

[0419] In some embodiments, the Fc-based chimeric protein complex comprises one or more linkers. In some embodiments, the Fc-based chimeric protein complex includes a linker that connects the Fc domain, antagonistic signaling agent, e.g. IL-6 antagonist(s) and targeting moiety(ies). In some embodiments, the Fc-based chimeric protein complex includes a linker that connects each antagonistic signaling agent, e.g. IL-6 antagonist, and

[0420] DB1 / 162518159.2 68 ORN-109PC / 114384-5109 targeting moiety (or, if more than one targeting moiety, an antagonistic signaling agent, e.g. IL-6 antagonist, to one of the targeting moieties). In some embodiments, the Fc-based chimeric protein complex includes a linker that connects each antagonistic signaling agent, e.g. IL-6 antagonist, to the Fc domain. In some embodiments, the Fc- based chimeric protein complex includes a linker that connects each targeting moiety to the Fc domain. In some embodiments, the Fc-based chimeric protein complex includes a linker that connects a targeting moiety to another targeting moiety. In some embodiments, the Fc-based chimeric protein complex includes a linker that connects an antagonistic signaling agent, e.g. IL-6 antagonist, to another signaling agent.

[0421] In some embodiments, a Fc-based chimeric protein complex comprises two or more targeting moieties. In such embodiments, the targeting moieties can be the same targeting moiety or they can be different targeting moieties.

[0422] In some embodiments, a Fc-based chimeric protein complex comprises two or more signaling agents In such embodiments, the signaling agents can be the same targeting moiety or they can be different targeting moieties.

[0423] By way of example, in some embodiments, the Fc-based chimeric protein complex comprise a Fc domain, at least two signaling agents (SA), and at least two targeting moieties (TM), wherein the Fc domain, signaling agents, and targeting moieties are selected from any of the Fc domains, signaling agents, and targeting moieties disclosed herein. In some embodiments, the Fc domain is homodimeric.

[0424] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 1A-F.

[0425] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 2A-H.

[0426] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 3A-H.

[0427] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 4A-D.

[0428] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 5A-F.

[0429] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 6A-J.

[0430] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 7A-D.

[0431] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 8A-F.

[0432] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs.

[0433] 9A-J.

[0434] DB1 / 162518159.2 69 ORN-109PC / 114384-5109

[0435] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 10A-F

[0436] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 11A-L.

[0437] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 12A-L.

[0438] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 13A-F

[0439] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 14A-L.

[0440] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 15A-L.

[0441] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 16A-J.

[0442] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 17A-J.

[0443] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 18A-F

[0444] In various embodiments, the Fc-based chimeric protein complex takes the form of any of the schematics of Figs. 19A-F

[0445] In some embodiments, the signaling agents are linked to the targeting moieties and the targeting moieties are linked to the Fc domain on the same terminus (see FIGs. 1A-F). In some embodiments, the Fc domain is homodimeric.

[0446] In some embodiments, the signaling agents and targeting moieties are linked to the Fc domain, wherein the targeting moieties and signaling agents are linked on the same terminus (see FIGs. 1A-F). In some embodiments, the Fc domain is homodimeric.

[0447] In some embodiments, the targeting moieties are linked to signaling agents and the signaling agents are linked to the Fc domain on the same terminus (see FIGs. 1A-F). In some embodiments, the Fc domain is homodimeric.

[0448] In some embodiments, the homodimeric Fc-based chimeric protein complex has two or more targeting moieties. In some embodiments, there are four targeting moieties and two signaling agents, the targeting moieties are linked to the Fc domain and the signaling agents are linked to targeting moieties on the same terminus (see FIGS. 2A- H). In some embodiments, the Fc domain is homodimeric. In some embodiments, where there are four targeting moieties and two signaling agents, two targeting moieties are linked to the Fc domain and two targeting moieties

[0449] DB1 / 162518159.2 70 ORN-109PC / 114384-5109 are linked to the signaling agents, which are linked to the Fc domain on the same terminus (see FIGS. 2A-H). In some embodiments, the Fc domain is homodimeric In some embodiments, where there are four targeting moieties and two signaling agents, two targeting moieties are linked to each other and one of the targeting moieties of from each pair is linked to the Fc domain on the same terminus and the signaling agents are linked to the Fc domain on the same terminus (see FIGS. 2A-H). In some embodiments, the Fc domain is homodimeric. In some embodiments, where there are four targeting moieties and two signaling agents, two targeting moieties are linked to each other, wherein one of the targeting moieties of from each pair is linked to an antagonistic signaling agent, e.g. IL-6 antagonist, and the other targeting moiety of the pair is linked the Fc domain, wherein the targeting moieties linked to the Fc domain are linked on the same terminus (see FIGS. 2A-H). In some embodiments, the Fc domain is homodimeric.

[0450] In some embodiments, the homodimeric Fc-based chimeric protein complex has two or more signaling agents. In some embodiments, where there are four signaling agents and two targeting moieties, two signaling agents are linked to each other and one of the signaling agents of from pair is linked to the Fc domain on the same terminus and the targeting moieties are linked to the Fc domain on the same terminus (see FIGs. 3A-H). In some embodiments, the Fc domain is homodimeric. In some embodiments, where there are four signaling agents and two targeting moieties, two signaling agents are linked to the Fc domain one the same terminus and two of the signaling agents are each linked to a targeting moiety, wherein the targeting moieties are linked to the Fc domain at the same terminus (see FIGs. 3A-H). In some embodiments, the Fc domain is homodimeric. In some embodiments, where there are four signaling agents and two targeting moieties, two signaling agents are linked to each other and one of the signaling agents of from pair is linked to a targeting moiety and the targeting moieties are linked to the Fc domain on the same terminus (see FIGs. 3A-H). In some embodiments, the Fc domain is homodimeric.

[0451] By way of example, in some embodiments, the Fc-based chimeric protein complex comprise a Fc domain, wherein the Fc domain comprises ionic pairing mutation(s) and / or knob-in-hole mutation(s), at least one antagonistic signaling agent, e.g. IL-6 antagonist, and at least one targeting moiety, wherein the ionic pairing motif and / or a knob-in-hole motif, antagonistic signaling agent, e.g. IL-6 antagonist, and targeting moiety are selected from any of the ionic pairing motif and / or a knob-in-hole motif, signaling agents, and targeting moieties disclosed herein. In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0452] In some embodiments, the antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the targeting moiety, which is linked to the Fc domain (see FIGs. 10A-F and 13A-F). In some embodiments, the targeting moiety is linked to the antagonistic signaling agent, e.g. IL-6 antagonist, which is linked to the Fc domain (see FIGs. 10A-F and 13A- F). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0453] DB1 / 162518159.2 71 ORN-109PC / 114384-5109

[0454] In some embodiments, the antagonistic signaling agent, e.g. IL-6 antagonist, and targeting moiety are linked to the Fc domain (see FIGs. 4A-D, 7A-D, 10A-F, and 13A-F). In some embodiments, the targeting moiety and the antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains on the same terminus (see FIGs. 4A-D and 7A-D). In some embodiments, the targeting moiety and the antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains on different termini (see FIGs. 4A-D and 7A-D). In some embodiments, the targeting moiety and the antagonistic signaling agent, e.g. IL-6 antagonist, are linked to the same Fc chain (see FIGs. 10A-F and 13A-F). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0455] In some embodiments, where there are one antagonistic signaling agent, e.g. IL-6 antagonist, and two targeting moieties, the antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the Fc domain and two targeting moieties can be: 1) linked to each other with one of the targeting moieties linked to the Fc domain; or 2) each linked to the Fc domain (see FIGs. 5A-F, 8A-F, 11A-L, 14A-L, 16A-J, and 17A-J). In some embodiments, the targeting moieties are linked on one Fc chain and the antagonistic signaling agent, e.g. IL-6 antagonist, is on the other Fc chain (see FIGs. 5A-F and 8A-F). In some embodiments, the paired targeting moieties and the antagonistic signaling agent, e.g. IL-6 antagonist, are linked to the same Fc chain (see FIGs 11A-L and 14A-L). In some embodiments, a targeting moiety is linked to the Fc domain and the other targeting moiety is linked to the antagonistic signaling agent, e.g. IL-6 antagonist, and the paired targeting moiety is linked to the Fc domain (see FIGs. 11A-L, 14A-L, 16A-J, and 17A-J). In some embodiments, the unpaired targeting moiety and paired targeting moiety are linked to the same Fc chain (see FIGs. 11A-L and 14A-L). In some embodiments, the unpaired targeting moiety and paired targeting moiety are linked to different Fc chains (see FIGs. 16A-J and 17A-J). In some embodiments, the unpaired targeting moiety and paired targeting moiety are linked on the same terminus (see FIGs. 16A-J and 17A-J). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0456] In some embodiments, where there are one antagonistic signaling agent, e.g. IL-6 antagonist, and two targeting moieties, a targeting moiety is linked to the antagonistic signaling agent, e.g. IL-6 antagonist, which is linked to the Fc domain, and the unpaired targeting moiety is linked the Fc domain (see FIGs. 11A-L, 14A-L, 16A-J, and 17A- J). In some embodiments, the paired antagonistic signaling agent, e.g. IL-6 antagonist, and unpaired targeting moiety are linked to the same Fc chain (see FIGs. 11 A-L and 14A-L). In some embodiments, the paired antagonistic signaling agent, e.g. IL-6 antagonist, and unpaired targeting moiety are linked to different Fc chains (see FIGs. 16A-J and 17A-J). In some embodiments, the paired antagonistic signaling agent, e.g. IL-6 antagonist, and unpaired targeting moiety are linked on the same terminus (see FIGs. 16A-J and 17A-J). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0457] In some embodiments, where there are one antagonistic signaling agent, e.g. IL-6 antagonist, and two targeting moieties, the targeting moieties are linked together and the antagonistic signaling agent, e.g. IL-6 antagonist, is linked to one of the paired targeting moieties, wherein the targeting moiety not linked to the antagonistic signaling

[0458] DB1 / 162518159.2 72 ORN-109PC / 114384-5109 agent, e.g. IL-6 antagonist, is linked to the Fc domain (see FIGs. 11 A-L and 14A-L). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0459] In some embodiments, where there are one antagonistic signaling agent, e.g. IL-6 antagonist, and two targeting moieties, the targeting moieties are linked together and the antagonistic signaling agent, e.g. IL-6 antagonist, is linked to one of the paired targeting moieties, wherein the antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the Fc domain (see FIGs. 11 A-L and 14A-L). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0460] In some embodiments, where there are one antagonistic signaling agent, e.g. IL-6 antagonist, and two targeting moieties, the targeting moieties are both linked to the antagonistic signaling agent, e.g. IL-6 antagonist, wherein one of the targeting moieties is linked to the Fc domain (see FIGs. 11 A-L and 14A-L). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0461] In some embodiments, where there are one antagonistic signaling agent, e.g. IL-6 antagonist, and two targeting moieties, the targeting moieties and the antagonistic signaling agent, e.g. IL-6 antagonist, are linked to the Fc domain (see FIGs. 16A-J and 17A-J). In some embodiments, the targeting moieties are linked on the terminus (see FIGs. 16A-J and 17A-J). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0462] In some embodiments, where there are two signaling agents and one targeting moiety, the signaling agents are linked to the Fc domain on the same terminus and the targeting moiety is linked to the Fc domain (see FIGs. 6A-J and 9A-J). In some embodiments, the signaling agents are linked to the Fc domain on the same Fc chain and the targeting moiety is linked on the other Fc chain (see FIGs. 18A-F and 19A-F). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0463] In some embodiments, where there are two signaling agents and one targeting moiety, an antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the targeting moiety, which is linked to the Fc domain and the other antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the Fc domain (see FIGs. 6A-J, 9A-J, 12A-L, and 15A-L). In some embodiments, the targeting moiety and the unpaired antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains (see FIGs. 6A-J and 9A-J) In some embodiments, the targeting moiety and the unpaired antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains on the same terminus (see FIGs. 6A-J and 9A-J) In some embodiments, the targeting moiety and the unpaired antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains on different termini (see FIGs. 6A-J and 9A- J). In some embodiments, the targeting moiety and the unpaired antagonistic signaling agent, e.g. IL-6 antagonist, are linked to the same Fc chains (see FIGs. 12A-L and 15A-L). In some embodiments, the Fc domain is

[0464] DB1 / 162518159.2 73 ORN-109PC / 114384-5109 heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0465] In some embodiments, where there are two signaling agents and one targeting moiety, the targeting moiety is linked to an antagonistic signaling agent, e.g. IL-6 antagonist, which is linked to the Fc domain and the other antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the Fc domain (see FIGs. 6A-J and 9A-J). In some embodiments, the paired antagonistic signaling agent, e.g. IL-6 antagonist, and the unpaired antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains (see FIGs 6A-J and 9A-J). In some embodiments, the paired antagonistic signaling agent, e.g. IL-6 antagonist, and the unpaired antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains on the same terminus (see FIGs. 6A-J and 9A-J). In some embodiments, the paired antagonistic signaling agent, e.g. IL-6 antagonist, and the unpaired antagonistic signaling agent, e.g. IL-6 antagonist, are linked to different Fc chains on different termini (see FIGs 6A-J and 9A-J). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0466] In some embodiments, where there are two signaling agents and one targeting moiety, the signaling agents are linked together and the targeting moiety is linked to one of the paired signaling agents, wherein the targeting moiety is linked to the Fc domain (see FIGs. 12A-L and 15A-L). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0467] In some embodiments, where there are two signaling agents and one targeting moiety, the signaling agents are linked together and one of the signaling agents is linked to the Fc domain and the targeting moiety is linked to the Fc domain (see FIGs. 12A-L, 15A-L, 18A-F, and 19A-F). In some embodiments, the paired signaling agents and targeting moiety are linked to the same Fc chain (see FIGs. 12A-L and 15A-L). In some embodiments, the paired signaling agents and targeting moiety are linked to different Fc chains (see FIGs 18A-F and 19A-F). In some embodiments, the paired signaling agents and targeting moiety are linked to different Fc chains on the same terminus (see FIGs. 18A-F and 19A-F). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0468] In some embodiments, where there are two signaling agents and one targeting moiety, the signaling agents are both linked to the targeting moiety, wherein one of the signaling agents is linked to the Fc domain (see FIGs. 12k- L and 15A-L). In some embodiments, the Fc domain is heterodimeric. In some embodiments, the Fc domain comprises a mutation that reduces or eliminates its effector function.

[0469] In some embodiments, where there are two signaling agents and one targeting moiety, the signaling agents are linked together and one of the signaling agents is linked to the targeting moiety and the other antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the Fc domain (see FIGs. 12A-L and 15A-L).

[0470] In some embodiments, where there are two signaling agents and one targeting moiety, each antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the Fc domain and the targeting moiety is linked to one of the signaling

[0471] DB1 / 162518159.2 74 ORN-109PC / 114384-5109 agents (see FIGs. 12A-L and 15A-L). In some embodiments, the signaling agents are linked to the same Fc chain (see FIGs. 12A-L and 15A-L).

[0472] In some embodiments, a targeting moiety or antagonistic signaling agent, e.g. IL-6 antagonist, is linked to the Fc domain, comprising one or both of CH2 and CH3 domains, and optionally a hinge region For example, vectors encoding the targeting moiety, antagonistic signaling agent, e.g. IL-6 antagonist, or combination thereof, linked as a single nucleotide sequence to an Fc domain can be used to prepare such polypeptides.

[0473] Linkers and Functional Groups

[0474] In some embodiments, the chimeric protein or chimeric protein complex optionally comprises one or more linkers. In some embodiments, the chimeric protein or chimeric protein complex comprise a linker connecting the targeting moiety and the antagonistic signaling agent (e.g., IL-6 antagonist). In some embodiments, the chimeric protein or chimeric protein complex comprise a linker within the antagonistic signaling agent (e.g., IL-6 antagonist). In some embodiments, the linker may be utilized to link various functional groups, residues, or moieties as described herein to the chimeric protein or chimeric protein complex. In some embodiments, the linker is a single amino acid or a plurality of amino acids that does not affect or reduce the stability, orientation, binding, neutralization, and / or clearance characteristics of the binding regions and the binding protein. In various embodiments, the linker is selected from a peptide, a protein, a sugar, or a nucleic acid.

[0475] In some embodiments, vectors encoding the chimeric protein or chimeric protein complex are linked as a single nucleotide sequence to any of the linkers described herein are provided and may be used to prepare such the chimeric protein or chimeric protein complex. In embodiments, the substituents of the Fc-based chimeric protein complex are expressed as nucleotide sequences in a vector

[0476] In some embodiments, the linker length allows for efficient binding of a targeting moiety and the antagonistic signaling agent (e.g., IL-6 antagonist) to their receptors. For instance, in some embodiments, the linker length allows for efficient binding of one of the targeting moieties and the signaling agent to receptors on the same cell.

[0477] In some embodiments, the linker length is at least equal to the minimum distance between the binding sites of one of the targeting moieties and the signaling agent to receptors on the same cell. In some embodiments the linker length is at least twice, or three times, or four times, or five times, or ten times, or twenty times, or 25 times, or 50 times, or one hundred times, or more the minimum distance between the binding sites of one of the targeting moieties and the signaling agent to receptors on the same cell

[0478] As described herein, the linker length allows for efficient binding of one of the targeting moieties and the signaling agent to receptors on the same cell, the binding being sequential, e.g. targeting moiety / receptor binding preceding signaling agent / receptor binding.

[0479] In some embodiments, there are two linkers in a single chimera, each connecting the signaling agent to a targeting moiety. In various embodiments, the linkers have lengths that allow for the formation of a site that has a disease cell and an effector cell without steric hindrance that would prevent modulation of the either cell.

[0480] DB1 / 162518159.2 75 ORN-109PC / 114384-5109

[0481] The invention contemplates the use of a variety of linker sequences. In various embodiments, the linker may be derived from naturally-occurring multi-domain proteins or are empirical linkers as described, for example, in Chichili ef al., (2013), Protein Sci. 22(2): 153-167, Chen et al., (2013), Adv Drug Deliv Rev. 65(10): 1357-1369, the entire contents of which are hereby incorporated by reference. In some embodiments, the linker may be designed using linker designing databases and computer programs such as those described in Chen et al., (2013), Adv Drug Deliv Rev. 65(10): 1357-1369 and Crasto et al., (2000), Protein Eng. 13(5):309-312, the entire contents of which are hereby incorporated by reference. In various embodiments, the linker may be functional For example, without limitation, the linker may function to improve the folding and / or stability, improve the expression, improve the pharmacokinetics, and / or improve the bioactivity of the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex

[0482] In some embodiments, the linker is a polypeptide. In some embodiments, the linker is less than about 100 amino acids long. For example, the linker may be less than about 100, about 95, about 90, about 85, about 80, about 75, about 70, about 65, about 60, about 55, about 50, about 45, about 40, about 35, about 30, about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 12, about 11 , about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, or about 2 amino acids long. In some embodiments, the linker is a polypeptide. In some embodiments, the linker is greater than about 100 amino acids long. For example, the linker may be greater than about 100, about 95, about 90, about 85, about 80, about 75, about 70, about 65, about 60, about 55, about 50, about 45, about 40, about 35, about 30, about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 12, about 11 , about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, or about 2 amino acids long. In some embodiments, the linker is flexible. In another embodiment, the linker is rigid.

[0483] In some embodiments directed to the chimeric protein or chimeric protein complex having two or more targeting moieties, a linker connects the two targeting moieties to each other and this linker has a short length and a linker connects a targeting moiety and a signaling agent this linker is longer than the linker connecting the two targeting moieties. For example, the difference in amino acid length between the linker connecting the two targeting moieties and the linker connecting a targeting moiety and a signaling agent may be about 100, about 95, about 90, about 85, about 80, about 75, about 70, about 65, about 60, about 55, about 50, about 45, about 40, about 35, about 30, about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 12, about 11, about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, or about 2 amino acids.

[0484] In some embodiments, the connector between the signaling moiety and the targeting moiety is a flexible linker. In various embodiments, the linker is substantially comprised of glycine and serine residues (e.g. about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90%, or about 95%, or about 97% glycines and serines). For example, in some embodiments, the linker is (Gly4Ser)n, where n is from about 1 to about 8, e.g. 1 , 2, 3, 4, 5, 6, 7, or 8 (SEQ ID NO:249 -SEQ ID NO:256, respectively). In an embodiment, the linker sequence is GGSGGSGGGGSGGGGS (SEQ ID NO:257). Additional illustrative linkers include, but are not limited to, linkers having the sequence LE, GGGGS (SEQ ID NO:249), (GGGGS)n(n=1-4) (SEQ ID NO:249 -SEQ ID

[0485] DB1 / 162518159.2 76 ORN-109PC / 114384-5109

[0486] NO:252). (Gly)8(SEQ ID NO:258), (Gly)6(SEQ ID NO:259), (EAAAK)n(n=1-3) (SEQ ID NO:260 -SEQ ID NO:262), A(EAAAK)nA (n = 2-5) (SEQ ID NO:263 - SEQ ID NO:266), AEAAAKEAAAKA (SEQ ID NO:263), A(EAAAK) ALEA(EAAAK)4A (SEQ ID NO:267), PAPAP (SEQ ID NO:268), KESGSVSSEQLAQFRSLD (SEQ ID NO:269), EGKSSGSGSESKST (SEQ ID NQ:270), GSAGSAAGSGEF (SEQ ID NO:271), and (XP)n, with X designating any amino acid, e.g., Ala, Lys, or Glu. In various embodiments, the linker is GGS or a repeat thereof wherein the GGS sequence is repeated 1 to 8 times (SEQ ID NO: 304 - 311). In some embodiments, the linker is GGGS or a repeat thereof wherein the GGGS sequence is repeated 1 to 8 times (SEQ ID NO: 312 - 319). In some embodiments, the linker has the sequence (GGS)n(n=9-20) (SEQ ID NO: 427-SEQ ID NO: 438).

[0487] In some embodiments, the linker is one or more of GGGSE (SEQ ID NO: 272), GSESG (SEQ ID NO: 273), GSEGS (SEQ ID NO: 274), GEGGSGEGSSGEGSSSEGGGSEGGGSEGGGSEGGS (SEQ ID NO: 275), and a linker of randomly placed G, S, and E every 4 amino acid intervals

[0488] In some embodiments, the linker is a synthetic linker such as PEG.

[0489] In various embodiments, the linker may be functional. For example, without limitation, the linker may function to improve the folding and / or stability, improve the expression, improve the pharmacokinetics, and / or improve the bioactivity of the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex. In another example, the linker may function to target the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex to a particular cell type or location.

[0490] In various embodiments, the present chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex may include one or more functional groups, residues, or moieties. In various embodiments, the one or more functional groups, residues, or moieties are attached or genetically fused to any of the signaling agents or targeting moieties described herein. In some embodiments, such functional groups, residues or moieties confer one or more desired properties or functionalities to the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex of the invention. Examples of such functional groups and of techniques for introducing them into the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex are known in the art, for example, see Remington's Pharmaceutical Sciences, 16th ed , Mack Publishing Co., Easton, Pa. (1980).

[0491] In various embodiments, each of the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex may by conjugated and / or fused with another agent to extend half-life or otherwise improve pharmacodynamic and pharmacokinetic properties. In some embodiments, the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex may be fused or conjugated with one or more of PEG, XTEN (e.g., as rPEG), polysialic acid (POLYXEN), albumin (e.g., human serum albumin or HAS), elastinlike protein (ELP), PAS, HAP, GLK, CTP, transferrin, and the like. In various embodiments, each of the individual chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex is fused to one or more of the agents described in BioDrugs (2015) 29:215-239, the entire contents of which are hereby incorporated by reference.

[0492] DB1 / 162518159.2 77 ORN-109PC / 114384-5109

[0493] In some embodiments, the functional groups, residues, or moieties comprise a suitable pharmacologically acceptable polymer, such as poly(ethyleneglycol) (PEG) or derivatives thereof (such as methoxypoly(ethyleneglycol) or mPEG). In some embodiments, attachment of the PEG moiety increases the halflife and / or reduces the immunogenecity of the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex. Generally, any suitable form of pegylation can be used, such as the pegylation used in the art for antibodies and antibody fragments (including but not limited to single domain antibodies such as VHHs); see, for example, Chapman, Nat. Biotechnol., 54, 531-545 (2002); by Veronese and Harris, Adv. Drug Deliv. Rev. 54, 453-456 (2003), by Harris and Chess, Nat. Rev. Drug. Discov., 2, (2003) and in WG04 / 060965, the entire contents of which are hereby incorporated by reference. Various reagents for pegylation of proteins are also commercially available, for example, from Nektar Therapeutics, USA. In some embodiments, site-directed pegylation is used, in particular via a cysteine-residue (see, for example, Yang et a / ., Protein Engineering, 16, 10, 761-770 (2003), the entire contents of which is hereby incorporated by reference). In some embodiments, the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex of the invention is modified so as to suitably introduce one or more cysteine residues for attachment of PEG, or an amino acid sequence comprising one or more cysteine residues for attachment of PEG may be fused to the amino-and / or carboxy-terminus of the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex, using techniques known in the art.

[0494] In some embodiments, the functional groups, residues, or moieties comprise N-linked or O-linked glycosylation. In some embodiments, the N-linked or O-linked glycosylation is introduced as part of a co-translational and / or post- translational modification

[0495] In some embodiments, the functional groups, residues, or moieties comprise one or more detectable labels or other signal-generating groups or moieties. Suitable labels and techniques for attaching, using and detecting them are known in the art and, include, but are not limited to, fluorescent labels (such as fluorescein, isothiocyanate, rhodamine, phycoerythrin, phycocyanin, allophycocyanin, o-phthaldehyde, and fluorescamine and fluorescent metals such as Eu or others metals from the lanthanide series), phosphorescent labels, chemiluminescent labels or bioluminescent labels (such as luminal, isoluminol, theromatic acridinium ester, imidazole, acridinium salts, oxalate ester, dioxetane or GFP and its analogs), radio-isotopes, metals, metals chelates or metallic cations or other metals or metallic cations that are particularly suited for use in in vivo, in vitro or in situ diagnosis and imaging, as well as chromophores and enzymes (such as malate dehydrogenase, staphylococcal nuclease, delta-V-steroid isomerase, yeast alcohol dehydrogenase, alpha-glycerophosphate dehydrogenase, triose phosphate isomerase, biotinavidin peroxidase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, betagalactosidase, ribonuclease, urease, catalase, glucose-VI-phosphate dehydrogenase, glucoamylase and acetylcholine esterase). Other suitable labels include moieties that can be detected using NMR or ESR spectroscopy. Such labeled VHHs and polypeptides of the invention may, for example, be used for in vitro, in vivo or in situ assays (including immunoassays known per se such as ELISA, RIA, EIA and other “sandwich assays,” etc.) as well as in vivo diagnostic and imaging purposes, depending on the choice of the specific label.

[0496] DB1 / 162518159.2 78 ORN-109PC / 114384-5109

[0497] In some embodiments, the functional groups, residues, or moieties comprise a tag that is attached or genetically fused to the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex. In some embodiments, the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex may include a single tag or multiple tags. The tag for example is a peptide, sugar, or DNA molecule that does not inhibit or prevent binding of the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex to its target or any other antigen of interest such as tumor antigens. In various embodiments, the tag is at least about: three to five amino acids long, five to eight amino acids long, eight to twelve amino acids long, twelve to fifteen amino acids long, or fifteen to twenty amino acids long. Illustrative tags are described for example, in U.S. Patent Publication No. US2013 / 0058962 In some embodiment, the tag is an affinity tag such as glutathione-S- transferase (GST) and histidine (His) tag. In an embodiment, the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex comprises a His tag.

[0498] In some embodiments, the functional groups, residues, or moieties comprise a chelating group, for example, to chelate one of the metals or metallic cations. Suitable chelating groups, for example, include, without limitation, diethyl-enetriaminepentaacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA).

[0499] In some embodiments, the functional groups, residues, or moieties comprise a functional group that is one part of a specific binding pair, such as the bioti n-(strept) avidi n binding pair. Such a functional group may be used to link the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex of the invention to another protein, polypeptide or chemical compound that is bound to the other half of the binding pair, / .e., through formation of the binding pair. For example, a chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex of the invention may be conjugated to biotin, and linked to another protein, polypeptide, compound or carrier conjugated to avidin or streptavidin For example, such a conjugated chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex may be used as a reporter, for example, in a diagnostic system where a detectable signal-producing agent is conjugated to avidin or streptavidin Such binding pairs may, for example, also be used to bind the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex to a carrier, including carriers suitable for pharmaceutical purposes. One nonlimiting example are the liposomal formulations described by Cao and Suresh, Journal of Drug Targeting, 8, 4, 257 (2000). Such binding pairs may also be used to link a therapeutically active agent to the chimeric proteins or chimeric protein complexes such as Fc-based chimeric protein complex of the invention.

[0500] Production of Chimeric Proteins or Chimeric Protein Complexes

[0501] Methods for producing the chimeric protein or chimeric protein complex of the invention are described herein. For example, DNA sequences encoding the chimeric protein or chimeric protein complex of the invention (e.g., DNA sequencesencoding the antagonistic signaling agent (e.g., IL-6 antagonist) and the targeting moiety and the linker) can be chemically synthesized using methods known in the art.

[0502] Synthetic DNA sequences can be ligated to other appropriate nucleotide sequences, including, e.g., expression control sequences, to produce gene expression constructs encoding the chimeric protein or chimeric protein

[0503] DB1 / 162518159.2 79 ORN-109PC / 114384-5109 complex. Accordingly, in various embodiments, the present invention provides for isolated nucleic acids comprising a nucleotide sequence encoding the chimeric protein or chimeric protein complex of the invention.

[0504] Nucleic acids encoding the chimeric protein or chimeric protein complex of the invention can be incorporated (ligated) into expression vectors, which can be introduced into host cells through transfection, transformation, or transduction techniques. For example, nucleic acids encoding the chimeric protein or chimeric protein complex of the invention can be introduced into host cells by retroviral transduction. Illustrative host cells are E. coli cells, Chinese hamster ovary (CHO) cells, human embryonic kidney 293 (HEK 293) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), and myeloma cells. Transformed host cells can be grown under conditions that permit the host cells to express the genes that encode the chimeric protein or chimeric protein complex of the invention. Accordingly, in various embodiments, the present invention provides expression vectors comprising nucleic acids that encode the chimeric protein or chimeric protein complex of the invention. In various embodiments, the present invention additional provides host cells comprising such expression vectors.

[0505] Specific expression and purification conditions will vary depending upon the expression system employed. For example, if a gene is to be expressed in E. coli, it is first cloned into an expression vector by positioning the engineered gene downstream from a suitable bacterial promoter, e.g., Trp or Tac, and a prokaryotic signal sequence. In another example, if the engineered gene is to be expressed in eukaryotic host cells, e.g., CHO cells, it is first inserted into an expression vector containing for example, a suitable eukaryotic promoter, a secretion signal, enhancers, and various introns. The gene construct can be introduced into the host cells using transfection, transformation, or transduction techniques.

[0506] The chimeric protein, the chimeric protein or chimeric protein complex of the invention can be produced by growing a host cell transfected with an expression vector encoding the chimeric protein or chimeric protein complex under conditions that permit expression of the protein. Following expression, the protein can be harvested and purified using techniques well known in the art, e.g., affinity tags such as glutathione-S-transferase (GST) and histidine tags or by chromatography.

[0507] Accordingly, in various embodiments, the present invention provides for a nucleic acid encoding a chimeric protein or chimeric protein complex of the present invention. In various embodiments, the present invention provides for a host cell comprising a nucleic acid encoding the chimeric protein or chimeric protein complex of the present invention.

[0508] In various embodiments, IL-6 antagonist, or the chimeric protein or chimeric protein complex comprising the IL-6 antagonist, may be expressed in vivo, for instance, in a patient. For example, in various embodiments, the IL-6 antagonist, or the chimeric protein or chimeric protein complex comprising the IL-6 antagonist, may be administered in the form of nucleic acid which encodes for the IL-6 antagonist or the chimeric protein or chimeric protein complex comprising IL-6 antagonist. In various embodiments, the nucleic acid is DNA or RNA. In some embodiments, the IL-6 antagonist, or the chimeric protein or chimeric protein complex comprising the IL-6

[0509] DB1 / 162518159.2 80 ORN-109PC / 114384-5109 antagonist, is encoded by a modified mRNA, i.e. an mRNA comprising one or more modified nucleotides. In some embodiments, the modified mRNA comprises one or modifications found in U.S. Patent No. 8,278,036, the entire contents of which are hereby incorporated by reference. In some embodiments, the modified mRNA comprises one or more of m5C, m5U, m6A, s2U, , and 2'-O-methyl-U. In some embodiments, the present invention relates to administering a modified mRNA encoding one or more of the chimeric protein or chimeric protein complex. In some embodiments, the present invention relates to gene therapy vectors comprising the same. In some embodiments, the present invention relates to gene therapy methods comprising the same. In various embodiments, the nucleic acid is in the form of an oncolytic virus, e.g. an adenovirus, reovirus, measles, herpes simplex, Newcastle disease virus or vaccinia.

[0510] In various embodiments, the chimeric protein or chimeric protein complex comprises a targeting moiety that is a VHH. In various embodiments, the VHH is not limited to a specific biological source or to a specific method of preparation. For example, the VHH can generally be obtained: (1) by isolating the HH domain of a naturally occurring heavy chain antibody; (2) by expression of a nucleotide sequence encoding a naturally occurring VHH domain; (3) by “humanization” of a naturally occurring VHH domain or by expression of a nucleic acid encoding a such humanized VHH domain; (4) by “camelization” of a naturally occurring VH domain from any animal species, such as from a mammalian species, such as from a human being, or by expression of a nucleic acid encoding such a camelized VH domain; (5) by “camelization” of a “domain antibody” or “Dab” as described in the art, or by expression of a nucleic acid encoding such a camelized VH domain; (6) by using synthetic or semi-synthetic techniques for preparing proteins, polypeptides or other amino acid sequences known in the art; (7) by preparing a nucleic acid encoding a VHH using techniques for nucleic acid synthesis known in the art, followed by expression of the nucleic acid thus obtained; and / or (8) by any combination of one or more of the foregoing.

[0511] In an embodiment, chimeric protein, the chimeric protein or chimeric protein complex comprises a VHH that corresponds to the VHH domains of naturally occurring heavy chain antibodies directed against a target of interest. In some embodiments, such VHH sequences can generally be generated or obtained by suitably immunizing a species of Camelid with a molecule of based on the target of interest (e.g., XCR1 , Clec9a, CD8, SIRPIa, FAP, etc.) (i.e., so as to raise an immune response and / or heavy chain antibodies directed against the target of interest), by obtaining a suitable biological sample from the Camelid (such as a blood sample, or any sample of B-cells), and by generating VHH sequences directed against the target of interest, starting from the sample, using any suitable known techniques. In some embodiments, naturally occurring VHH domains against the target of interest can be obtained from naive libraries of Camelid VHH sequences, for example, by screening such a library using the target of interest or at least one part, fragment, antigenic determinant or epitope thereof using one or more screening techniques known in the art. Such libraries and techniques are, for example, described in WO 9937681 , WO 0190190, WO 03025020 and WO 03035694, the entire contents of which are hereby incorporated by reference. In some embodiments, improved synthetic or semi-synthetic libraries derived from naive VHH libraries may be used, such as VHH libraries obtained from naive VHH libraries by techniques such as random mutagenesis and / or CDR shuffling, as for example, described in WO 0043507, the entire contents of which are hereby incorporated by

[0512] DB1 / 162518159.2 81 ORN-109PC / 114384-5109 reference. In some embodiments, another technique for obtaining V H sequences directed against a target of interest involves suitably immunizing a transgenic mammal that is capable of expressing heavy chain antibodies ( / .e., so as to raise an immune response and / or heavy chain antibodies directed against the target of interest), obtaining a suitable biological sample from the transgenic mammal (such as a blood sample, or any sample of B- cells), and then generating VHH sequences directed against XCR1 starting from the sample, using any suitable known techniques. For example, for this purpose, the heavy chain antibody-expressing mice and the further methods and techniques described in WO 02085945 and in WO 04049794 (the entire contents of which are hereby incorporated by reference) can be used.

[0513] In an embodiment, the chimeric protein or chimeric protein complex of the present invention comprises a VHH that has been “humanized” / .e., by replacing one or more amino acid residues in the amino acid sequence of the naturally occurring V H sequence (and in particular in the framework sequences) by one or more of the amino acid residues that occur at the corresponding position(s) in a VH domain from a conventional 4-chain antibody from a human being. This can be performed using humanization techniques known in the art. In some embodiments, possible humanizing substitutions or combinations of humanizing substitutions may be determined by methods known in the art, for example, by a comparison between the sequence of a VHH and the sequence of a naturally occurring human VH domain. In some embodiments, the humanizing substitutions are chosen such that the resulting humanized VHHs still retain advantageous functional properties. Generally, as a result of humanization, the VHHs of the invention may become more “human-like,” while still retaining favorable properties such as a reduced immunogenicity, compared to the corresponding naturally occurring VHH domains. In various embodiments, the humanized VHHs of the invention can be obtained in any suitable manner known in the art and thus are not strictly limited to polypeptides that have been obtained using a polypeptide that comprises a naturally occurring VHH domain as a starting material.

[0514] In an embodiment, the chimeric protein or chimeric protein complex comprises a VHH that has been “camelized,” / .e., by replacing one or more amino acid residues in the amino acid sequence of a naturally occurring VH domain from a conventional 4-chain antibody by one or more of the amino acid residues that occur at the corresponding position(s) in a VHH domain of a heavy chain antibody of a camelid. In some embodiments, such “camelizing” substitutions are inserted at amino acid positions that form and / or are present at the VH-VL interface, and / or at the so-called Camelidae hallmark residues (see, for example, WO9404678, the entire contents of which are hereby incorporated by reference). In some embodiments, the VH sequence that is used as a starting material or starting point for generating or designing the camelized VHH is a VH sequence from a mammal, for example, the VH sequence of a human being, such as a VH3 sequence. In various embodiments, the camelized VHHs can be obtained in any suitable manner known in the art ( / .e., as indicated under points (1)-(8) above) and thus are not strictly limited to polypeptides that have been obtained using a polypeptide that comprises a naturally occurring VH domain as a starting material.

[0515] In various embodiments, both “humanization” and “camelization” can be performed by providing a nucleotide sequence that encodes a naturally occurring VHH domain or VH domain, respectively, and then changing, in a

[0516] DB1 / 162518159.2 82 ORN-109PC / 114384-5109 manner known in the art, one or more codons in the nucleotide sequence in such a way that the new nucleotide sequence encodes a “humanized” or “camelized” VHH, respectively. This nucleic acid can then be expressed in a manner known in the art, so as to provide the desired VHH of the invention. Alternatively, based on the amino acid sequence of a naturally occurring V H domain or H domain, respectively, the amino acid sequence of the desired humanized or camelized VHH of the invention, respectively, can be designed and then synthesized de novo using techniques for peptide synthesis known in the art Also, based on the amino acid sequence or nucleotide sequence of a naturally occurring VHH domain or VH domain, respectively, a nucleotide sequence encoding the desired humanized or camelized VHH, respectively, can be designed and then synthesized de novo using techniques for nucleic acid synthesis known in the art, after which the nucleic acid thus obtained can be expressed in a manner known in the art, so as to provide the desired VHH of the invention. Other suitable methods and techniques for obtaining the VHHs of the invention and / or nucleic acids encoding the same, starting from naturally occurring VH sequences or VHH sequences, are known in the art, and may, for example, comprise combining one or more parts of one or more naturally occurring VH sequences (such as one or more FR sequences and / or CDR sequences), one or more parts of one or more naturally occurring VHH sequences (such as one or more FR sequences or CDR sequences), and / or one or more synthetic or semi-synthetic sequences, in a suitable manner, so as to provide a VHH of the invention or a nucleotide sequence or nucleic acid encoding the same.

[0517] Pharmaceutically Acceptable Salts and Excipients

[0518] Chimeric proteins or chimeric protein complexes described herein can possess a sufficiently basic functional group, which can react with an inorganic or organic acid, or a carboxyl group, which can react with an inorganic or organic base, to form a pharmaceutically acceptable salt. A pharmaceutically acceptable acid addition salt is formed from a pharmaceutically acceptable acid, as is well known in the art. Such salts include the pharmaceutically acceptable salts listed in, for example, Journal of Pharmaceutical Science, 66, 2-19 (1977) and The Handbook of Pharmaceutical Salts; Properties, Selection, and Use P. H. Stahl and C. G. Wermuth (eds.), Verlag, Zurich (Switzerland) 2002, which are hereby incorporated by reference in their entirety.

[0519] Pharmaceutically acceptable salts include, by way of non-limiting example, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, pamoate, phenylacetate, trifluoroacetate, acrylate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, isobutyrate, phenylbutyrate, a-hydroxybutyrate, butyne-1,4-dicarboxylate, hexyne-1 ,4- dicarboxylate, caprate, caprylate, cinnamate, glycollate, heptanoate, hippurate, malate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate, phthalate, teraphthalate, propiolate, propionate, phenylpropionate, sebacate, suberate, p-bromobenzenesulfonate, chlorobenzenesulfonate, ethylsulfonate, 2-hydroxyethylsulfonate, methylsulfonate, naphthalene-1 -sulfonate, naphthalene-2-sulfonate, naphthalene-1 ,5-sulfonate, xylenesulfonate, and tartarate salts.

[0520] DB1 / 162518159.2 83 ORN-109PC / 114384-5109

[0521] The term “pharmaceutically acceptable salt” also refers to a salt of the compositions of the present invention having an acidic functional group, such as a carboxylic acid functional group, and a base. Suitable bases include, but are not limited to, hydroxides of alkali metals such as sodium, potassium, and lithium; hydroxides of alkaline earth metal such as calcium and magnesium; hydroxides of other metals, such as aluminum and zinc; ammonia, and organic amines, such as unsubstituted or hydroxy-substituted mono-, di-, or tri-alkylamines, dicyclohexylamine; tributyl amine; pyridine; N-methyl, N-ethylamine; diethylamine; triethylamine; mono-, bis-, or tris-(2-OH-lower alkylamines), such as mono-; bis-, or tris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butylamine, or tris- (hydroxymethyl)methylamine, N,N-di-lower alkyl-N-(hydroxyl-lower alkylj-amines, such as N,N-dimethyl-N-(2- hydroxyethyl)amine or tri-(2-hydroxyethyl)am ine; N-methyl-D-glucamine; and amino acids such as arginine, lysine, and the like.

[0522] In some embodiments, the compositions described herein are in the form of a pharmaceutically acceptable salt.

[0523] Pharmaceutical Compositions and Formulations

[0524] In various embodiments, the present invention pertains to pharmaceutical compositions comprising the chimeric protein or chimeric protein complex described herein and a pharmaceutically acceptable carrier or excipient. Any pharmaceutical compositions described herein can be administered to a subject as a component of a composition that comprises a pharmaceutically acceptable carrier or vehicle. Such compositions can optionally comprise a suitable amount of a pharmaceutically acceptable excipient so as to provide the form for proper administration.

[0525] In various embodiments, pharmaceutical excipients can be liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. The pharmaceutical excipients can be, for example, saline, gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, urea and the like. In addition, auxiliary, stabilizing, thickening, lubricating, and coloring agents can be used. In one embodiment, the pharmaceutically acceptable excipients are sterile when administered to a subject. Water is a useful excipient when any agent described herein is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid excipients, specifically for injectable solutions. Suitable pharmaceutical excipients also include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. Any agent described herein, if desired, can also comprise minor amounts of wetting or emulsifying agents, or pH buffering agents. Other examples of suitable pharmaceutical excipients are described in Remington’s Pharmaceutical Sciences 1447-1676 (Alfonso R. Gennaro eds., 19th ed. 1995), incorporated herein by reference.

[0526] The present invention includes the described pharmaceutical compositions (and / or additional therapeutic agents) in various formulations. Any inventive pharmaceutical composition (and / or additional therapeutic agents) described herein can take the form of solutions, suspensions, emulsion, drops, tablets, pills, pellets, capsules, capsules containing liquids, gelatin capsules, powders, sustained-release formulations, suppositories, emulsions, aerosols, sprays, suspensions, lyophilized powder, frozen suspension, desiccated powder, or any other form suitable for

[0527] DB1 / 162518159.2 84 ORN-109PC / 114384-5109 use. In one embodiment, the composition is in the form of a capsule. In another embodiment, the composition is in the form of a tablet. In yet another embodiment, the pharmaceutical composition is formulated in the form of a soft-gel capsule. In a further embodiment, the pharmaceutical composition is formulated in the form of a gelatin capsule. In yet another embodiment, the pharmaceutical composition is formulated as a liquid.

[0528] Where necessary, the inventive pharmaceutical compositions (and / or additional agents) can also include a solubilizing agent. Also, the agents can be delivered with a suitable vehicle or delivery device as known in the art. Combination therapies outlined herein can be co-delivered in a single delivery vehicle or delivery device.

[0529] The formulations comprising the inventive pharmaceutical compositions (and / or additional agents) of the present invention may conveniently be presented in unit dosage forms and may be prepared by any of the methods well known in the art of pharmacy. Such methods generally include the step of bringing the therapeutic agents into association with a carrier, which constitutes one or more accessory ingredients. Typically, the formulations are prepared by uniformly and intimately bringing the therapeutic agent into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product into dosage forms of the desired formulation (e.g., wet or dry granulation, powder blends, etc., followed by tableting using conventional methods known in the art).

[0530] In various embodiments, any pharmaceutical compositions (and / or additional agents) described herein is formulated in accordance with routine procedures as a composition adapted for a mode of administration described herein

[0531] Routes of administration include, for example: oral, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, to the lung, by inhalation, sublingual, intranasal, intracerebral, intravaginal, transdermal, rectally, by inhalation, or topically. Administration can be local or systemic. In some embodiments, the administering is effected orally. In another embodiment, the administration is by parenteral injection or via aerosol or nebulizer. The mode of administration can be left to the discretion of the practitioner, and depends inpart upon the site of the medical condition. In most instances, administration results in the release of any agent described herein into the bloodstream In some embodiments, the administration is via liquid nebulization, dry powder dispersion or meter-dose administration.

[0532] In one embodiment, the chimeric protein or chimeric protein complex described herein is formulated in accordance with routine procedures as a composition adapted for oral administration. Compositions for oral delivery can be in the form of tablets, lozenges, aqueous or oily suspensions, granules, powders, emulsions, capsules, syrups, or elixirs, for example. Orally administered compositions can comprise one or more agents, for example, sweetening agents such as fructose, aspartame or saccharin; flavoring agents such as peppermint, oil of Wintergreen, or cherry; coloring agents; and preserving agents, to provide a pharmaceutically palatable preparation. Moreover, where in tablet or pill form, the compositions can be coated to delay disintegration and absorption in the gastrointestinal tract thereby providing a sustained action over an extended period of time. Selectively permeable membranes surrounding an osmotically active driving any chimeric protein or chimeric protein complex described herein are

[0533] DB1 / 162518159.2 85 ORN-109PC / 114384-5109 also suitable for orally administered compositions. In these latter platforms, fluid from the environment surrounding the capsule is imbibed by the driving compound, which swells to displace the agent or agent composition through an aperture. These delivery platforms can provide an essentially zero order delivery profile as opposed to the spiked profiles of immediate release formulations. A time-delay material such as glycerol monostearate or glycerol stearate can also be useful Oral compositions can include standard excipients such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, and magnesium carbonate. In one embodiment, the excipients are of pharmaceutical grade. Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, tragacanth, etc., and mixtures thereof.

[0534] Dosage forms suitable for parenteral administration (e.g. intravenous, intramuscular, intraperitoneal, subcutaneous and intra-articular injection and infusion) include, for example, solutions, suspensions, dispersions, emulsions, and the like. They may also be manufactured in the form of sterile solid compositions (e.g. lyophilized composition), which can be dissolved or suspended in sterile injectable medium immediately before use. They may contain, for example, suspending or dispersing agents known in the art. Formulation components suitable for parenteral administration include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as EDTA; buffers such as acetates, citrates or phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose.

[0535] For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). The carrier should be stable under the conditions of manufacture and storage, and should be preserved against microorganisms. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol), and suitable mixtures thereof.

[0536] The compositions provided herein, alone or in combination with other suitable components, can be made into aerosol formulations ( / .e., “nebulized”) to be administered via inhalation. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like.

[0537] Any inventive pharmaceutical compositions (and / or additional agents) described herein can be administered by controlled-release or sustained-release means or by delivery devices that are well known to those of ordinary skill in the art. Examples include, but are not limited to, those described in U.S. Patent Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591 ,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; and 5,733,556, each of which is incorporated herein by reference in its entirety. Such dosage forms can be useful for providing controlled-or sustained-release of one or more active ingredients using, for example, hydropropyl cellulose, hydropropylmethyl cellulose, polyvinylpyrrolidone, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or a combination thereof to provide the desired release profile in varying proportions. Suitable controlled- or sustained-release formulations known to

[0538] DB1 / 162518159.2 86 ORN-109PC / 114384-5109 those skilled in the art, including those described herein, can be readily selected for use with the active ingredients of the agents described herein. The invention thus provides single unit dosage forms suitable for oral administration such as, but not limited to, tablets, capsules, gelcaps, and caplets that are adapted for controlled- or sustained- release.

[0539] Controlled- or sustained-release of an active ingredient can be stimulated by various conditions, including but not limited to, changes in pH, changes in temperature, stimulation by an appropriate wavelength of light, concentration or availability of enzymes, concentration or availability of water, or other physiological conditions or compounds.

[0540] In another embodiment, a controlled-release system can be placed in proximity of the target area to be treated, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Other controlled-release systems discussed in the review by Langer, 1990, Science 249:1527-1533) may be used.

[0541] Pharmaceutical formulations preferably are sterile. Sterilization can be accomplished, for example, by filtration through sterile filtration membranes. Where the composition is lyophilized, filter sterilization can be conducted prior to or following lyophilization and reconstitution.

[0542] Administration and Dosage

[0543] It will be appreciated that the actual dose of the chimeric protein or chimeric protein complex to be administered according to the present invention will vary according to the particular dosage form, and the mode of administration. Many factors that may modify the action of the chimeric protein or chimeric protein complex (e.g., body weight, gender, diet, time of administration, route of administration, rate of excretion, condition of the subject, drug combinations, genetic disposition and reaction sensitivities) can be taken into account by those skilled in the art. Administration can be carried out continuously or in one or more discrete doses within the maximum tolerated dose. Optimal administration rates for a given set of conditions can be ascertained by those skilled in the art using conventional dosage administration tests.

[0544] In some embodiments, a suitable dosage of the chimeric protein or chimeric protein complex is in a range of about 0.01 pig / kg to about 100 mg / kg of body weight of the subject, about 0.01 pig / kg to about 10 mg / kg of body weight of the subject, or about 0.01 pig / kg to about 1 mg / kg of body weight of the subject for example, about 0.01 pig / kg, about 0.02 pig / kg, about 0.03 pig / kg, about 0.04 pig / kg, about 0.05 pig / kg, about 0.06 pig / kg, about 0.07 pig / kg, about 0.08 pig / kg, about 0.09 pig / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1 .1 mg / kg, about 1.2 mg / kg, about 1 3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1 .7 mg / kg, about 1.8 mg / kg, 1.9 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg body weight, or about 100 mg / kg body weight, inclusive of all values and ranges therebetween.

[0545] DB1 / 162518159.2 87 ORN-109PC / 114384-5109

[0546] Individual doses of the chimeric protein or chimeric protein complex can be administered in unit dosage forms (e.g., tablets, capsules, or liquid formulations) containing, for example, from about 1 pig to about 100 mg, from about 1 pig to about 90 mg, from about 1 pig to about 80 mg, from about 1 pig to about 70 mg, from about 1 pg to about 60 mg, from about 1 pg to about 50 mg, from about 1 pg to about 40 mg, from about 1 pg to about 30 mg, from about 1 pg to about 20 mg, from about 1 pg to about 10 mg, from about 1 pg to about 5 mg, from about 1 pg to about 3 mg, from about 1 pg to about 1 mg per unit dosage form, or from about 1 pg to about 50 pg per unit dosage form. For example, a unit dosage form can be about 1 pg, about 2 pg, about 3 pg, about 4 pg, about 5 pg, about 6 pg, about / pg, about 8 pg, about 9 pg, about 10 pg, about 11 pg, about 12 pg, about 13 pg, about 14 pg, about 15 pg, about 16 pg, about 17 pg, about 18 pg, about 19 pg, about 20 pg, about 21 pg, about 22 pg, about 23 pg, about 24 pg, about 25 pg, about 26 pg, about 27 pg, about 28 pg, about 29, about 30 pg, about 35 pg, about 40 pg, about 45 pg, about 50 pg, about 60 pg, about 70 pg, about 80 pg, about 90 pg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg, inclusive of all values and ranges therebetween.

[0547] In one embodiment, the chimeric protein or chimeric protein complex is administered at an amount of from about 1 pg to about 100 mg daily, from about 1 pg to about 90 mg daily, from about 1 pg to about 80 mg daily, from about 1 pg to about 70 mg daily, from about 1 pg to about 60 mg daily, from about 1 pg to about 50 mg daily, from about 1 pg to about 40 mg daily, from about 1 pg to about 30 mg daily, from about 1 pg to about 20 mg daily, from about 01 pg to about 10 mg daily, from about 1 pg to about 5 mg daily, from about 1 pg to about 3 mg daily, or from about 1 pg to about 1 mg daily. In various embodiments, the chimeric protein or chimeric protein complex is administered at a daily dose of about 1 pg, about 2 pg, about 3 pg, about 4 pg, about 5 pg, about 6 pg, about 7 pg, about 8 pg, about 9 pg, about 10 pg, about 11 pg, about 12 pg, about 13 pg, about 14 pg, about 15 pg, about 16 pg, about 17 pg, about 18 pg, about 19 pg, about 20 pg,, about 21 pg, about 22 pg, about 23 pg, about 24 pg, about 25 pg, about 26 pg, about 27 pg, about 28 pg, about 29, about 30 pg, about 35 pg, about 40 pg, about 45 pg, about 50 pg, about 60 pg, about 70 pg, about 80 pg, about 90 pg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, about 05 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg, inclusive of all values and ranges therebetween.

[0548] Combination Therapy and Additional Therapeutic Agents

[0549] In various embodiments, the pharmaceutical composition of the present invention is co-administered in conjunction with additional therapeutic agent(s). Co-administration can be simultaneous or sequential.

[0550] DB1 / 162518159.2 88 ORN-109PC / 114384-5109

[0551] In one embodiment, the additional therapeutic agent and chimeric protein or chimeric protein complex of the present invention are administered to a subject simultaneously. The term “simultaneously” as used herein, means that the additional therapeutic agent and chimeric protein or chimeric protein complex are administered with a time separation of no more than about 60 minutes, such as no more than about 30 minutes, no more than about 20 minutes, no more than about 10 minutes, no more than about 5 minutes, or no more than about 1 minute. Administration of the additional therapeutic agent and chimeric protein or chimeric protein complex can be by simultaneous administration of a single formulation (e.g., a formulation comprising the additional therapeutic agent and chimeric protein or chimeric protein complex) or of separate formulations (e.g., a first formulation including the additional therapeutic agent and a second formulation including chimeric protein or chimeric protein complex).

[0552] Co-administration does not require the therapeutic agents to be administered simultaneously, if the timing of their administration is such that the pharmacological activities of the additional therapeutic agent and chimeric protein or chimeric protein complex overlap in time, thereby exerting a combined therapeutic effect. For example, the additional therapeutic agent and chimeric protein or chimeric protein complex can be administered sequentially. The term “sequentially” as used herein means that the additional therapeutic agent and chimeric protein or chimeric protein complex are administered with a time separation of more than about 60 minutes. For example, the time between the sequential administration of the additional therapeutic agent and chimeric protein or chimeric protein complex can be more than about 60 minutes, more than about 2 hours, more than about 5 hours, more than about 10 hours, more than about 1 day, more than about 2 days, more than about 3 days, more than about 1 week apart, more than about 2 weeks apart, or more than about one month apart. The optimal administration times will depend on the rates of metabolism, excretion, and / or the pharmacodynamic activity of the additional therapeutic agent and chimeric protein or chimeric protein complex being administered. Either the additional therapeutic agent or chimeric protein or chimeric protein complex cell may be administered first.

[0553] Co-administration also does not require the therapeutic agents to be administered to the subject by the same route of administration. Rather, each therapeutic agent can be administered by any appropriate route, for example, parenterally or non-parenterally.

[0554] In some embodiments, chimeric protein or chimeric protein complex described herein acts synergistically when coadministered with another therapeutic agent. In such embodiments, chimeric protein or chimeric protein complex and the additional therapeutic agent may be administered at doses that are lower than the doses employed when the agents are used in the context of monotherapy

[0555] In some embodiments, the present invention pertains to chemotherapeutic agents as additional therapeutic agents. For example, without limitation, such combination of chimeric protein or chimeric protein complex and chemotherapeutic agent find use in the treatment of cancers, as described elsewhere herein. Examples of chemotherapeutic agents include, but are not limited to, alkylating agents such as thiotepa and CYTOXAN cydosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine,

[0556] DB1 / 162518159.2 89 ORN-109PC / 114384-5109 triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (e.g., bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; cally statin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (e.g., cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB 1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammall and calicheamicin omegall (see, e.g., Agnew, Chem. Inti. Ed. Engl., 33: 183-186 (1994)); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN doxorubicin (including morpholino- doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxy doxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as minoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2, 2', 2"- trichlorotriethylamine; trichothecenes (e.g., T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g., TAXOL paclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J ), ABRAXANE Cremophor-free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, 111.), and TAXOTERE doxetaxel (Rhone-Poulenc Rorer, Antony, France); chloranbucil; GEMZAR gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE, vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (Camptosar, CPT-11 ) (including the treatment regimen of irinotecan with 5-FU and leucovorin);

[0557] DB1 / 162518159.2 90 ORN-109PC / 114384-5109 topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; combretastatin; leucovorin (LV); oxaliplatin, including the oxaliplatin treatment regimen (FOLFOX); lapatinib (Tykerb); inhibitors of PKC-ci, Raf, H-Ras, EGFR (e.g., erlotinib (Tarceva)) and VEGF-A that reduce cell proliferation and pharmaceutically acceptable salts, acids or derivatives of any of the above. In addition, the methods of treatment can further include the use of radiation In addition, the methods of treatment can further include the use of photodynamic therapy.

[0558] In some embodiments, the present invention relates to combination therapies using chimeric protein or chimeric protein complex and an immunosuppressive agent. In some embodiments, the present invention relates to administration of the Clec9A binding agent to a patient undergoing treatment with an immunosuppressive agent.

[0559] In an embodiment, the immunosuppressive agent is TNF. In illustrative embodiments, chimeric protein or chimeric protein complex act synergistically when co-administered with TNF. In an illustrative embodiment, chimeric protein or chimeric protein complex act synergistically when co-administered with TNF for use in treating tumor or cancer. For example, co-administration of chimeric protein or chimeric protein complex of the present invention and TNF may act synergistically to reduce or eliminate the tumor or cancer, or slow the growth and / or progression and / or metastasis of the tumor or cancer. In some embodiments, the combination of chimeric protein or chimeric protein complex and TNF may exhibit improved safety profiles when compared to the agents used alone in the context of monotherapy In some embodiments, chimeric protein or chimeric protein complex and TNF may be administered at doses that are lower than the doses employed when the agents are used in the context of monotherapy.

[0560] In some embodiments, inclusive, without limitation, of autoimmune applications, the additional therapeutic agent is an immunosuppressive agent that is an anti-inflammatory agent such as a steroidal anti-inflammatory agent or a non-steroidal anti-inflammatory agent (NSAID). Steroids, particularly the adrenal corticosteroids and their synthetic analogues, are well known in the art. Examples of corticosteroids useful in the present invention include, without limitation, hydroxyltriamcinolone, alpha-methyl dexamethasone, beta-methyl betamethasone, beclomethasone dipropionate, betamethasone benzoate, betamethasone dipropionate, betamethasone valerate, dobetasol valerate, desonide, desoxymethasone, dexamethasone, diflorasone diacetate, diflucortolone valerate, fluadrenolone, fluclorolone acetonide, flumethasone pivalate, fluosinolone acetonide, fluocinonide, flucortine butylester, fluocortolone, fluprednidene (fluprednylidene) acetate, flurandrenolone, halcinonide, hydrocortisone acetate, hydrocortisone butyrate, methylprednisolone, triamcinolone acetonide, cortisone, cortodoxone, flucetonide, fludrocortisone, difluorosone diacetate, fluradrenolone acetonide, medrysone, amcinafel, amcinafide, betamethasone and the balance of its esters, chloroprednisone, clocortelone, clescinolone, dichlorisone, difluprednate, flucloronide, flunisolide, fluoromethalone, fluperolone, fluprednisolone, hydrocortisone, meprednisone, paramethasone, prednisolone, prednisone, beclomethasone dipropionate. (NSAIDS) that may be used in the present invention, include but are not limited to, salicylic acid, acetyl salicylic acid, methyl salicylate, glycol salicylate, salicylmides, benzyl-2,5-diacetoxybenzoic acid, ibuprofen, fulindac, naproxen, ketoprofen, etofenamate, phenylbutazone, and indomethacin. In some embodiments, the immunosupressive agent may be cytostatics such as alkylating agents, antimetabolites (e.g., azathioprine, methotrexate), cytotoxic antibiotics,

[0561] DB1 / 162518159.2 91 ORN-109PC / 114384-5109 antibodies (e.g., basiliximab, daclizumab, and muromonab), anti-immunophilins (e.g., cyclosporine, tacrolimus, sirolimus), inteferons, opioids, TNF binding proteins, mycophenolates, and small biological agents (e.g., fingolimod, myriocin). Additional anti-inflammatory agents are described, for example, in U.S. Patent No. 4,537,776, the entire contents of which is incorporated by reference herein.

[0562] In some embodiments, the present invention pertains to various agents used for treating obesity as additional therapeutic agents. Illustrative agents used for treating obesity include, but are not limited to, orlistat (e.g. ALL1 , XENICAL), loracaserin (e.g. BELVIQ), phentermine-topiramate (e.g. QSYMIA), sibutramme (e.g. REDUCTIL or MERJDIA), rimonabant (ACOMPLLA), exenatide (e.g. BYETTA), pramlintide (e.g. SYMLIN) phentermine, benzphetamine, diethylpropion, phendimetrazme, bupropion, and metformin. Agents that interfere with the body's ability to absorb specific nutrients in food are among the additional agents, e.g. orlistat (e.g. ALU, XENICAL), glucomannan, and guar gum. Agents that suppress appetite are also among the additional agents, e.g. catecholamines and their derivatives (such as phenteimine and other amphetamine-based drugs), various antidepressants and mood stabilizers (e.g. bupropion and topiramate), anorectics (e.g. dexedrine, digoxin). Agents that increase the body's metabolism are also among the additional agents

[0563] In some embodiments, additional therapeutic agents may be selected from among appetite suppressants, neurotransmitter reuptake inhibitors, dopaminergic agonists, serotonergic agonists, modulators of GABAergic signaling, anticonvulsants, antidepressants, monoamine oxidase inhibitors, substance P (NK1) receptor antagonists, melanocortin receptor agonists and antagonists, lipase inhibitors, inhibitors of fat absorption, regulators of energy intake or metabolism, cannabinoid receptor modulators, agents for treating addiction, agents for treating metabolic syndrome, peroxisome proliferator-activated receptor (PPAR) modulators; dipeptidyl peptidase 4 (DPP- 4) antagonists, agents for treating cardiovascular disease, agents for treating elevated triglyceride levels, agents for treating low HDL, agents for treating hypercholesterolemia, and agents for treating hypertension. Some agents for cardiovascular disease include statins (e.g. lovastatin, atorvastatin, fluvastatin, rosuvastatin, simvastatin and pravastatin) and omega-3 agents (e.g. LOVAZA, EPANQVA, VASCEPA, esterified omega-3's in general, fish oils, krill oils, algal oils). In some embodiments, additional agents may be selected from among amphetamines, benzodiazepines, suifonyl ureas, meglitinides, thiazolidinediones, biguanides, betablockers, XCE inhibitors, diuretics, nitrates, calcium channel blockers, phenlermine, sibutramine, iorcaserin, cetilistat, rimonabant, taranabant, topiramate, gabapentin, valproate, vigabatrin, bupropion, tiagabine, sertraline, fluoxetine, trazodone, zonisamide, methylphenidate, varenicline, naltrexone, diethylpropion, phendimetrazine, rcpaglini.de, nateglinide, glimepiride, metformin, pioglitazone, rosiglilazone, and sitagliptin.

[0564] In some embodiments, the present invention pertains to an agent used for treating diabetes as additional therapeutic agents. Illustrative anti-diabetic agents include, but are not limited to, sulfonylurea (e.g., DYMELOR (acetohexamide), DIABINESE (chlorpropamide), ORINASE (tolbutamide), and TOLINASE (tolazamide), GLUCOTROL (glipizide), GLUCOTROL XL (extended release), DIABETA (glyburide), MICRONASE (glyburide), GLYNASE PRESTAB (glyburide), and AMARYL (glimepiride)); a Biguanide (e.g. metformin (GLUCOPHAGE, GLUCOPHAGE XR, RIOMET, FORTAMET, and GLUMETZA)); a thiazolidinedione (e.g. ACTOS (pioglitazone)

[0565] DB1 / 162518159.2 92 ORN-109PC / 114384-5109 and AVANDIA (rosiglitazone); an alpha-glucosidase inhibitor (e.g., PRECOSE (acarbose) and GLYSET (miglitol); a Meglitinide (e.g., PRANDIN (repaglinide) and STARLIX (nateglinide)); a Dipeptidyl peptidase IV (DPP-IV) inhibitor (e.g., JANUVIA (sitagliptin), NESINA (alogliptin), ONGLYZA (saxagliptin), and TRADJENTA (linagliptin)); Sodium-glucose co-transporter 2 (SGLT2) inhibitor (e.g. INVOKANA (canaglifozin)); and a combination pill (e.g. GLUCOVANCE, which combines glyburide (a sulfonylurea) and metformin, METAGLIP, which combines glipizide (a sulfonylurea) and metformin, and AVANDAMET, which uses both metformin and rosiglitazone (AVANDIA) in one pill, KAZANO (alogliptin and metformin), OSENI (alogliptin plus pioglitazone), METFORMIN oral, ACTOS oral, BYETTA subcutaneous, JANUVIA oral, WELCHOL oral, JANUMET oral, glipizide oral, glimepiride oral, GLUCOPHAGE oral, LANTUS subcutaneous, glyburide oral, ONGLYZA oral, AMARYI oral, LANTUS SOLOSTAR subcutaneous, BYDUREON subcutaneous, LEVEMIR FLEXPEN subcutaneous, ACTOPLUS MET oral, GLUMETZA oral, TRADJENTA oral, bromocriptine oral, KOMBIGLYZE XR oral, INVOKANA oral, PRANDIN oral, LEVEMIR subcutaneous, PARLODEL oral, pioglitazone oral, NOVOLOG subcutaneous, NOVOLOG FLEXPEN subcutaneous, VICTOZA 2-PAK subcutaneous, HUMALOG subcutaneous, STARLIX oral, FORTAMET oral, GLUCOVANCE oral, GLUCOPHAGE XR oral, NOVOLOG Mix 70-30 FLEXPEN subcutaneous, GLYBURIDE- METFORMIN oral, acarbose oral, SYMUNPEN 60 subcutaneous, GLUCOTROI XL oral, NOVOLIN R inj, GLUCOTROL oral, DUETACT oral, sitagliptin oral, SYMLINPEN 120 subcutaneous, HUMALOG KWIKPEN subcutaneous, JANUMET XR oral, GLIPIZIDE-METFORMIN oral, CYCLOSET oral, HUMALOG MIX 75-25 subcutaneous, nateglinide oral, HUMALOG Mix 75-25 KWIKPEN subcutaneous, HUMULIN 70 / 30 subcutaneous, PRECOSE oral, APIDRA subcutaneous, Humulin R inj, Jentadueto oral, Victoza 3-Pak subcutaneous, Novolin 70 / 30 subcutaneous, NOVOLIN N subcutaneous, insulin detemir subcutaneous, glyburide micronized oral, GLYNASE oral, HUMULIN N subcutaneous, insulin glargine subcutaneous, RIOMET oral, pioglitazone-metformin oral, APIDRA SOLOSTAR subcutaneous, insulin lispro subcutaneous, GLYSET oral, HUMULIN 70 / 30 Pen subcutaneous, colesevelam oral, sitagliptin-metformin oral, DIABETA oral, insulin regular human inj, HUMULIN N Pen subcutaneous, exenatide subcutaneous, HUMALOG Mix 50-50 KWIKPEN subcutaneous, liraglutide subcutaneous, KAZANO oral, repaglinide oral, chlorpropamide oral, insulin aspart subcutaneous, NOVOLOG Mix 70-30 subcutaneous, HUMALOG Mix 50-50 subcutaneous, saxagliptin oral, ACTOPLUS MetXR oral, miglitol oral, NPH insulin human recomb subcutaneous, insulin NPH and regular human subcutaneous, tolazamide oral, mifepristone oral, insulin aspart protam-insulin aspart subcutaneous, repaglinide-metformin oral, saxagliptin- metformin oral, linagliptin-metformin oral, NESINA oral, OSENI oral, tolbutamide oral, insulin lispro protamine and lispro subcutaneous, pramlintide subcutaneous, insulin glulisine subcutaneous, pioglitazone-glimepiride oral, PRANDIMET oral, NOVOLOG PenFill subcutaneous, linagliptin oral, exenatide microspheres subcutaneous, KORLYM oral, alogliptin oral, alogliptin-pioglitazone oral, alogliptin-metformin oral, canagliflozin oral, Lispro (HUMALOG); Aspart (NOVOLOG); Glulisine (APIDRA); Regular (NOVOLIN R or HUMULIN R); NPH (NOVOLIN N or HUMULIN N); Glargine (LANTUS); Detemir (LEVEMIR); HUMULIN or NOVOLIN 70 / 30; and NOVOLOG Mix 70 / 30 HUMALOG Mix 75 / 25 or 50 / 50.

[0566] DB1 / 162518159.2 93 ORN-109PC / 114384-5109

[0567] In some embodiments, chimeric protein or chimeric protein complex of the present invention act synergistically when used in combination with Chimeric Antigen Receptor (CAR) T-cell therapy. In an illustrative embodiment, chimeric protein or chimeric protein complex act synergistically when used in combination with CAR T-cell therapy in treating tumor or cancer. In an embodiment, chimeric protein or chimeric protein complex act synergistically when used in combination with CAR T-cell therapy in treating blood-based tumors. In an embodiment, chimeric protein or chimeric protein complex act synergistically when used in combination with CAR T-cell therapy in treating solid tumors. For example, use of chimeric protein or chimeric protein complex and CAR T-cells may act synergistically to reduce or eliminate the tumor or cancer, or slow the growth and / or progression and / or metastasis of the tumor or cancer. In various embodiments, chimeric protein or chimeric protein complex of the invention induces CAR T-cell division. In various embodiments, chimeric protein or chimeric protein complex of the invention induces CAR T-cell proliferation. In various embodiments, chimeric protein or chimeric protein complex of the invention prevents anergy of the CAR T cells.

[0568] In various embodiments, the CAR T-cell therapy comprises CAR T cells that target antigens (e.g., tumor antigens) such as, but not limited to, carbonic anhydrase IX (CAIX), 5T4, CD19, CD20, CD22, CD30, CD33, CD38, CD47, CS1, CD138, Lewis-Y, L1-CAM, MUC16, ROR-1, IL13Ro2, gp100, prostate stem cell antigen (PSCA), prostatespecific membrane antigen (PSMA), B-cell maturation antigen (BCMA), human papillomavirus type 16 E6 (HPV- 16 E6), CD171 , folate receptor alpha (FR-a), GD2, human epidermal growth factor receptor 2 (HER2), mesothelin, EGFRvlll, fibroblast activation protein (FAP), carcinoembryonic antigen (CEA), and vascular endothelial growth factor receptor 2 (VEGF-R2), as well as other tumor antigens well known in the art. Additional illustrative tumor antigens include, but are not limited to MART-1 / Melan-A, gp100, Dipeptidyl peptidase IV (DPPIV), adenosine deaminase-binding prote...

Claims

ORN-109PC / 114384-5109CLAIMSWhat is claimed is:

1. A chimeric protein or protein complex comprising:(a) an interleukin-6 (IL-6) antagonist comprising an amino acid sequence having at least about 90% identity with SEQ ID NO: 426, and one or more mutations at an amino acid position selected from V121 , Y31, G35, L57, E59, N60, S118, and W157, and(b) one or more targeting moieties, the targeting moieties comprising recognition domains which specifically bind to an antigen or receptor of interest; wherein the IL-6 antagonist and the one or more targeting moieties are optionally connected with one or more linkers, wherein the one or more mutations of the IL-6 antagonist confer ablated activity and / or affinity for IL-6 receptor (gp130).

2. The chimeric protein or protein complex of claim 1 , wherein the IL-6 antagonist comprises an amino acid sequence having at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity with SEQ ID NO: 426.

3. The chimeric protein or protein complex of claim 1 or 2, wherein the mutation comprises a hydrophilic or hydrophobic amino acid residue.

4. The chimeric protein or protein complex of any one of the above claims, wherein the hydrophilic amino acid residue is a polar and positively charged hydrophilic residue selected from arginine (R) and lysine (K) or an aromatic, polar and positively charged hydrophilic including histidine (H).

5. The chimeric protein or protein complex of any one of claims 1-3, wherein the hydrophilic amino acid residue is a polar and neutral of charge hydrophilic residue selected from asparagine (N), glutamine (Q), serine (S), threonine (T), proline (P), and cysteine (C).

6. The chimeric protein or protein complex of any one of claims 1-3, wherein the hydrophilic amino acid residue is a polar and negatively charged hydrophilic residue selected from aspartate (D) and glutamate (E).

7. The chimeric protein or protein complex of any one of claims 1-3, wherein the hydrophobic amino acid is a hydrophobic, aliphatic amino acid selected from glycine (G), alanine (A), leucine (L), isoleucine (I), methionine (M), and valine (V) or a hydrophobic, aromatic amino acid selected from phenylalanine (F), tryptophan (W), and tyrosine (Y).

8. The chimeric protein or protein complex of any one of claims 1-7, wherein the one or more mutations of the IL-6 antagonist is selected from V121 D, Y31 D, Y31 F, Y31G, G35F, L57A, L57D, E59F, N60W, S118R, W157A, W157G, V121 D S118R, Y31 G_L57A_V121 D_W157A, and Y31G_L57A_S118R_V121 D_W157A, where the residue numbering is based on SEQ ID NO: 426,126DBl / 162518159.2ORN-109PC / 114384-5109 optionally wherein the mutation is V121D, and the IL-6 antagonist further comprises one or more mutations selected from Y31G, L57A, S118R, W157A, and W157G.

9. The chimeric protein or protein complex of any one of claims 1-8, wherein the one or more mutations confer ablated activity and / or affinity that is inducible by attachment to one or more targeting moieties.

10. The chimeric protein or protein complex of any one of claims 1-9, wherein the targeting moiety is directed against a tumor cell.

11. The chimeric protein orprotein complexof any one of claims 1-10, wherein the targeting moiety comprises a recognition domain that recognizes and / or binds an antigen or receptor on a tumor cell, endothelial cell, epithelial cell, mesenchymal cell, tumor stroma or adipose stromal cell, ECM and / or immune cell, organ cells, and / or tissue cells.

12. The chimeric protein or protein complex of claim 11, wherein the immune cell is selected from a T cell, a B cell, a dendritic cell, a macrophage, a neutrophil, a mast cell, a monocyte, a red blood cell, myeloid cell, myeloid derived suppressor cell, a NKT cell, and a NK cell, or derivatives thereof.

13. The chimeric protein or protein complex of any one of claims 1-12, wherein the targeting moiety comprises a recognition domain that is a recombinant heavy-chain-only antibody (VHH), a full-length antibody or a fragment thereof, a single-domain antibody, a single-chain antibody (scFv), a Humabody, a shark heavy-chain-only antibody (VNAR), a microprotein (e.g. cysteine knot protein, knottin), a darpin, an anticalin, an adnectin, an aptamer, a Fv, a Fab, a Fab', a F(ab')2, a peptide mimetic molecule, a natural ligand for a receptor, or a synthetic molecule.

14. The chimeric protein or protein complex of claim 13, wherein the recognition domain is a single-domain antibody or a single-chain antibody (scFv).

15. The chimeric protein or protein complex of claim 14, wherein the single domain antibody is a recombinant heavy-chain-only antibody (VHH) or humanized VHH16. The chimeric protein or protein complex of any one of claims 1-15, wherein the recognition domain functionally modulates the antigen or receptor of interest.

17. The chimeric protein or protein complex of any one of claims 1-15, wherein the recognition domain binds but does not functionally modulate the antigen or receptor of interest.

18. The chimeric protein or protein complex of any one of claims 1-17, comprising two or more targeting moieties.

19. The chimeric protein or protein complex of any one of the above claims, wherein the targeting moiety binds to one or more of the follow targets: CD8, CTLA4, CD3, CD4, DNAM-1 , Nrp1 (neurophilin), TNFR2, GITR, ICOS, CD20, CD70, Clec9a, NKp46, asialoglycoprotein receptor (ASGPR), PD-1 , PD-L1 , PD-L2, SIRPIa, FAP, XCR1, tenascin, ASC, prostate-specific membrane antigen (PSMA), or ECM proteins.DBl / 162518159.2 127ORN-109PC / 114384-510920. The chimeric protein or protein complex of any one of claims 1-19, wherein the connector between (a) and (b) is a flexible linker.21 . The chimeric protein or protein complex of claim 20, wherein the flexible linker is substantially comprised of glycine and serine residues, optionally wherein i) the flexible linker comprises (Gly4Ser)n, where n is from about 1 to about 8, or ii) the flexible linker comprises one or more of SEQ ID NOs: 304-319.

22. The chimeric protein or protein complex of claim 20, wherein the flexible linker is or comprises GGSGGSGGGGSGGGGS (SEQ ID NO: 257).

23. The chimeric protein or protein complex of claim 20, wherein the flexible linker is or comprises LE, GGGGS (SEQ ID NO:249), (GGGGS)n (n=1-4) (SEQ ID NO:249-SEQ ID NO:252), (Gly)8 (SEQ ID NO:258), (Gly)6 (SEQ ID NO:259), (EAAAK)n (n=1-3) (SEQ ID NQ:260-SEQ ID NO:262), A(EAAAK)nA (n = 2-5) (SEQ ID NO:263- SEQ ID NO:266), AEAAAKEAAAKA (SEQ ID NO:263), A(EAAAK)4ALEA(EAAAK)4A (SEQ ID NO:267), PAPAP (SEQ ID NO:268), KESGSVSSEQLAQFRSLD (SEQ ID NO:269), EGKSSGSGSESKST (SEQ ID NQ:270), GSAGSAAGSGEF (SEQ ID NO:271), (GGS)n(n=1-8) (SEQ ID NO: 304-311), (GGGS)n(n=1-8) (SEQ ID NO: 312- 319), (GGS)n (n=9-20) (SEQ ID NO: 427-SEQ ID NO: 438), and (XP)n, with X designating any amino acid, e.g., Ala, Lys, or Glu24. The chimeric protein or protein complex of any one of claims 1 -23, wherein the chimeric protein or protein complex is suitable for use in a patient having one or more of: autoimmune diseases or disorders, immune disorders, cancer, hepatocellular carcinoma, pancreatic cancer / melanoma, obesity, Graft-versus-host disease (GVHD), viral infections, cardiovascular diseases, wound, ischemia-related diseases, neurodegenerative diseases, and / or metabolic diseases25. A recombinant nucleic acid encoding one or more chimeric proteins or protein complexes of any one of claims 1-23.

26. A host cell comprising the nucleic acid of claim 25.

27. A pharmaceutical composition comprising one or more chimeric proteins or protein complexes of any one of claims 1-23 and a pharmaceutical excipient28. A method for treating an autoimmune disease or disorder, comprising administering an effective amount of i) the chimeric protein or protein complex of any one of the claims 1-23; ii) the recombinant nucleic acid of claim 25; Hi) the host cell of claim 26; or (iv) the pharmaceutical composition of claim 27 to a patient in need thereof29. The method of claim 28, wherein the autoimmune disease or disorder is selected from multiple sclerosis, diabetes mellitus, lupus, celiac disease, Crohn's disease, ulcerative colitis, Guillain-Barre syndrome, scleroderms, Goodpasture's syndrome, Wegener's granulomatosis, autoimmune epilepsy, Rasmussen's encephalitis, Primary biliary sclerosis, Sclerosing cholangitis, Autoimmune hepatitis, Addison's disease, Hashimoto's thyroiditis, Fibromyalgia, Menier's syndrome; transplantation rejection (e.g., prevention of allograft rejection) pernicious anemia, rheumatoid arthritis, systemic lupus erythematosus, dermatomyositis, Sjogren's syndrome, lupusDBl / 162518159.2 128ORN-109PC / 114384-5109 erythematosus, multiple sclerosis, myasthenia gravis, Reiter's syndrome, Grave's disease, and systemic lupus erythematosis.

30. The method of claim 29, wherein the autoimmune disease or disorder is multiple sclerosis.31 . A method for treating cancer, comprising administering an effective amount of i) the chimeric protein or protein complex of any one of the claims 1 -23; ii) the recombinant nucleic acid of claim 25; iii) the host cell of claim 26; or (iv) the pharmaceutical composition of claim 27, to a patient in need thereof.

32. The method of claim 31, wherein the cancer is selected form one or more of myeloma, multiple myeloma, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system cancer, breast cancer, cancer of the peritoneum, cervical cancer, choriocarcinoma, colon and rectum cancer, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, cancer of the head and neck, gastric cancer, gastrointestinal cancer, glioblastoma, hepatic carcinoma, hepatoma, intra-epithelial neoplasm, kidney or renal cancer, larynx cancer, leukemia, liver cancer, lung cancer, small-cell lung cancer, nonsmall cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, melanoma, neuroblastoma, oral cavity cancer, ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer, cancer of the respiratory system, salivary gland carcinoma, sarcoma, Kaposi’s sarcoma, skin cancer, squamous cell cancer, stomach cancer, testicular cancer, thyroid cancer, uterine or endometrial cancer, cancer of the urinary system, vulval cancer, lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, B-cell lymphoma, low grade / follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate grade / follicular NHL, intermediate grade diffuse NHL, high grade immunoblastic NHL, high grade lymphoblastic NHL, high grade small non-cleaved cell NHL, bulky disease NHL, mantle cell lymphoma, AIDS-related lymphoma, Waldenstrom's Macroglobulinemia, chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia as well as other carcinomas and sarcomas, post-transplant lymphoproliferative disorder (PTLD), abnormal vascular proliferation associated with phakomatoses, edema, and Meigs' syndrome.

33. The method of claim 32, wherein the cancer is multiple myeloma.

34. The chimeric protein or protein complex of any one of claims 1-23 or the pharmaceutical composition of claim Tl for use as a medicament.

35. Use of the chimeric protein or protein complex of any one of claims 1-23 or the pharmaceutical composition of claim 27 in the manufacture of a medicament.

36. The chimeric protein or protein complex of any one of claims 1-23 or the pharmaceutical composition of claim 27 for use in the treatment of autoimmune diseases or disorders, immune disorders, cancer, hepatocellular carcinoma, pancreatic cancer / melanoma, obesity, Graft-versus-host disease (GVHD), viral infections, cardiovascular diseases, wound, ischemia-related diseases, neurodegenerative diseases, and / or metabolic diseases.

37. A Fc-based chimeric protein complex comprising:DBl / 162518159.2 129ORN-109PC / 114384-5109(a) an interleukin-6 (IL-6) antagonist comprising an amino acid sequence having at least about 90% identity with SEQ ID NO: 426, and one or more mutations at an amino acid position selected from V121 , Y31, G35, L57, E59, N60, S118, and W157, and(b) one or more targeting moieties, said targeting moieties comprising recognition domains which specifically bind to an antigen or receptor of interest; and(c) a Fc domain, the Fc domain optionally having one or more mutations that reduces or eliminates one or more effector functions of the Fc domain, promotes Fc chain pairing in the Fc domain, and / or stabilizes a hinge region in the Fc domain, wherein the one or more mutations of the IL-6 antagonist confer ablated affinity and / or activity for IL-6 receptor (gp130).

38. The Fc-based chimeric protein complex of claim 37, wherein the IL-6 antagonist comprises an amino acid sequence having at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity with SEQ ID NO: 426.

39. The Fc-based chimeric protein complex of claim 37 or 38, wherein the mutation comprises a hydrophilic or hydrophobic amino acid residue.

40. The Fc-based chimeric protein complex of any one of claims 37-39, wherein the hydrophilic amino acid residue is a polar and positively charged hydrophilic residue selected from arginine (R) and lysine (K) or an aromatic, polar and positively charged hydrophilic including histidine (H).41 . The Fc-based chimeric protein complex of any one of claims 37-39, wherein the hydrophilic amino acid residue is a polar and neutral of charge hydrophilic residue selected from asparagine (N), glutamine (Q), serine (S), threonine (T), proline (P), and cysteine (C).

42. The Fc-based chimeric protein complex of any one of claims 37-39, wherein the hydrophilic amino acid residue is a polar and negatively charged hydrophilic residue selected from aspartate (D) and glutamate (E).

43. The Fc-based chimeric protein complex of any one of claims 37-39, wherein the hydrophobic amino acid is a hydrophobic, aliphatic amino acid selected from glycine (G), alanine (A), leucine (L), isoleucine (I), methionine (M), and valine (V) or a hydrophobic, aromatic amino acid selected from phenylalanine (F), tryptophan (W), and tyrosine (Y).

44. The Fc-based chimeric protein complex of any one of claims 37-43, wherein the one or more mutations of the IL-6 antagonist is selected from V121 D, Y31 D, Y31 F, Y31G, G35F, L57A, L57D, E59F, N60W, S118R, W157A, W157G, V121D S118R, Y31G_L57A_V121D_W157A, and Y31G_L57A_S118R_V121D_W157A, where the residue numbering is based on SEQ ID NO: 426, optionally wherein the mutation is V121D, and the IL-6 antagonist further comprises one or more mutations selected from Y31G, L57A, S118R, W157A, and W157G.DBl / 162518159.2 130ORN-109PC / 114384-510945. The Fc-based chimeric protein complex of any one of claims 37-44, wherein the one or more mutations confer ablated affinity and / or activity that is inducible by attachment to one or more targeting moieties or upon inclusion in the Fc-based chimeric protein complex.

46. The Fc-based chimeric protein complex of any one of 37-45, wherein the targeting moiety is directed against a tumor cell.

47. The Fc-based chimeric protein complex of any one of 37-45, wherein the targeting moiety comprises a recognition domain that recognizes and / or binds an antigen or receptor on a tumor cell, endothelial cell, epithelial cell, mesenchymal cell, tumor stroma or adipose stromal cell, ECM and / or immune cell, organ cells, and / or tissue cells.

48. The Fc-based chimeric protein complex of claim 47, wherein the immune cell is selected from a T cell, a B cell, a dendritic cell, a macrophage, a neutrophil, a mast cell, a monocyte, a red blood cell, myeloid cell, myeloid derived suppressor cell, a NKT cell, and a NK cell, or derivatives thereof.

49. The Fc-based chimeric protein complex of any one of claims 37-48, wherein the targeting moiety comprises a recognition domain that is a recombinant heavy-chain-only antibody (VHH), a single-domain antibody, a single-chain antibody (scFv), a shark heavy-chain-only antibody (VNAR), a microprotein (e.g. cysteine knot protein, knottin), a darpin, an anticalin, an adnectin, an aptamer, a Fv, a Fab, a Fab', a F(ab')2, a peptide mimetic molecule, a natural ligand for a receptor, or a synthetic molecule.

50. The Fc-based chimeric protein complex of any one of claims 37-49, wherein the recognition domain is a single-domain antibody, optionally a VHH or a humanized VHH.51 . The Fc-based chimeric protein complex of any one of claims 37-50, wherein the recognition domain functionally modulates the antigen or receptor of interest.

52. The Fc-based chimeric protein complex of any one of claims 37-51 , wherein the recognition domain binds but does not functionally modulate the antigen or receptor of interest.

53. The Fc-based chimeric protein complex of any one of claims 37-52, comprising two or more targeting moieties.

54. The Fc-based chimeric protein complex of any one of claims 37-53, wherein the targeting moiety binds to one or more of the follow targets: CDS, CTLA4, CD3, CD4, DNAM-1 , Nrp1 (neurophilin), TNFR2, GITR, ICOS, CD20, CD70, Clec9a, NKp46, asialoglycoprotein receptor (ASGPR), PD-1 , PD-L1, PD-L2, SIRPIa, FAP, XCR1 , tenascin, prostate-specific membrane antigen (PSMA), or ECM proteins.

55. A recombinant nucleic acid encoding one or more Fc-based chimeric protein complexes of any one of claims 37-54, or a constituent polypeptide thereof.

56. A host cell comprising the nucleic acid of claim 55.DBl / 162518159.2 131ORN-109PC / 114384-510957. A pharmaceutical composition comprising one or more one or more Fc-based chimeric protein complexes of any one of claims 37-54 and a pharmaceutical excipient.

58. A method for treating an autoimmune disease or disorder, comprising administering an effective amount of i) the Fc-based chimeric protein of any one of the claims 37-54; ii) the recombinant nucleic acid of claim 55; Hi) the host cell of claim 56; or (iv) the pharmaceutical composition of claim 57, to a patient in need thereof.

59. The method of claim 58, wherein the autoimmune disease or disorder is selected from multiple sclerosis, diabetes mellitus, lupus, celiac disease, Crohn's disease, ulcerative colitis, Guillain-Barre syndrome, scleroderms, Goodpasture's syndrome, Wegener's granulomatosis, autoimmune epilepsy, Rasmussen's encephalitis, Primary biliary sclerosis, Sclerosing cholangitis, Autoimmune hepatitis, Addison's disease, Hashimoto's thyroiditis, Fibromyalgia, Menier's syndrome; transplantation rejection (e.g., prevention of allograft rejection) pernicious anemia, rheumatoid arthritis, systemic lupus erythematosus, dermatomyositis, Sjogren's syndrome, lupus erythematosus, multiple sclerosis, myasthenia gravis, Reiter's syndrome, Grave's disease, and systemic lupus erythematosis.

60. The method of claim 59, wherein the autoimmune disease or disorder is multiple sclerosis.61 . A method for treating cancer, comprising administering an effective amount of i) the Fc-based chimeric protein of any one of the claims 37-54; ii) the recombinant nucleic acid of claim 55; iii) the host cell of claim 56; or (iv) the pharmaceutical composition of claim 57, to a patient in need thereof.

62. The method of claim 61, wherein the cancer is selected form one or more of myeloma, multiple myeloma, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system cancer, breast cancer, cancer of the peritoneum, cervical cancer, choriocarcinoma, colon and rectum cancer, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, cancer of the head and neck, gastric cancer, gastrointestinal cancer, glioblastoma, hepatic carcinoma, hepatoma, intra-epithelial neoplasm, kidney or renal cancer, larynx cancer, leukemia, liver cancer, lung cancer, small-cell lung cancer, nonsmall cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, melanoma, neuroblastoma, oral cavity cancer, ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer, cancer of the respiratory system, salivary gland carcinoma, sarcoma, Kaposi’s sarcoma, skin cancer, squamous cell cancer, stomach cancer, testicular cancer, thyroid cancer, uterine or endometrial cancer, cancer of the urinary system, vulval cancer, lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, B-cell lymphoma, low grade / follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate grade / follicular NHL, intermediate grade diffuse NHL, high grade immunoblastic NHL, high grade lymphoblastic NHL, high grade small non-cleaved cell NHL, bulky disease NHL, mantle cell lymphoma, AIDS-related lymphoma, Waldenstrom's Macroglobulinemia, chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia as well as other carcinomas and sarcomas, post-transplant lymphoproliferative disorder (PTLD), abnormal vascular proliferation associated with phakomatoses, edema, and Meigs' syndrome.

63. The method of claim 62, wherein the cancer is multiple myeloma.DBl / 162518159.2 132ORN-109PC / 114384-510964. The Fc-based chimeric protein complex of any one of claims 37-54 or the pharmaceutical composition of claim 57 for use as a medicament.

65. Use of the Fc-based chimeric protein complex of any one of claims 37-54 or the pharmaceutical composition of claim 57 in the manufacture of a medicament.

66. The Fc-based chimeric protein complex of any one of claims 37-54 or the pharmaceutical composition of claim 57 for use in the treatment of autoimmune diseases or disorders, immune disorders, cancer, hepatocellular carcinoma, pancreatic cancer / melanoma, obesity, Graft-versus-host disease (GVHD), viral infections, cardiovascular diseases, wound, ischemia-related diseases, neurodegenerative diseases, and / or metabolic diseases.

67. The Fc-based chimeric protein complex of any one of claims 37-54, wherein the Fc domain is from IgG, IgA, IgD, IgM or IgE.

68. The Fc-based chimeric protein complex of claim 67, wherein the IgG is selected from IgG 1 , lgG2, lgG3, or lgG4.

69. The Fc-based chimeric protein complex of any one of claims 37-54, wherein the Fc domain is from human IgG, IgA, IgD, IgM or IgE70. The Fc-based chimeric protein complex of claim 69, wherein the human IgG is selected from human lgG1 , lgG2, lgG3, or lgG4.

71. The Fc-based chimeric protein complex of any one of claims 37-54 or 67-70, wherein the Fc chain pairing is promoted by ionic pairing and / or a knob-in-hole pairing.

72. The Fc-based chimeric protein complex of any one of claims 37-54 or 67-71 , wherein the one or more mutations to the Fc domain results in an ionic pairing between the Fc chains in the Fc domain.

73. The Fc-based chimeric protein complex of any one of claims 37-54 or 67-72, wherein the one or more mutations to the Fc domain results in a knob-in-hole pairing in the Fc domain.

74. The Fc-based chimeric protein complex of any one of claims 37-54 or 67-73, wherein the one or more mutations to the Fc domain results in the reduction or elimination of an effector function of the Fc domain.

75. The Fc based chimeric protein complex of any one of claims 37-54 or 67-74, wherein the Fc-based chimeric protein complex is a heterodimer and has a trans orientation / configuration, as relates to any targeting moiety and signaling agent, relative to each other, or any targeting moieties relative to each other, or any signaling agents relative to each other.

76. The Fc based chimeric protein complex of any one of claims 37-54 or 67-75, wherein the Fc-based chimeric protein complex is a heterodimer and has a cis orientation, as relates to any targeting moiety and signaling agent, relative to each other, or any targeting moieties relative to each other, or any signaling agents relative to each other.DBl / 162518159.2 133ORN-109PC / 114384-510977. The Fc based chimeric protein complex of any one of claims 37-54 or 67-76, wherein the Fc comprises L234A, L235A, and K322Q substitutions in human IgG 1 (according to EU numbering).

78. The Fc-based chimeric protein complex of any one of claims 37-54 or 67-77, wherein the Fc is human lgG1 , and optionally contains one or more mutations of L234, L235, K322, D265, P329, and P331 (according to EU numbering).

79. The Fc-based chimeric protein complex of any one of claims 37-54 or 67-78, wherein the Fc-based chimeric protein complex has an orientation and / or configuration of any one of FIGs 1A-F, 2A-H, 3A-H, 4A-D, 5A- F, 6A-J, 7A-D, 8A-F, 9A-J, 10A-F, 11A-L, 12A-L, 13A-F, 14A-L, 15A-L, 16A-J, 17A-J, 18A-F, and 19A-F.

80. The chimeric protein or protein complex of any one of claims 1-23 or the Fc-based chimeric protein complex of any one of claims 37-54 or 67-79, wherein the chimeric protein or protein complex or Fc-based chimeric protein complex comprises a polypeptide comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity with one of SEQ ID NO: 447, 414-425, 439-446, 448-456.81 . The chimeric protein or protein complex or the Fc-based chimeric protein complex of claim 80, wherein the chimeric protein or Fc-based chimeric protein complex comprises a polypeptide having an amino acid sequence selected from SEQ ID NO: 447, 414-425, 439-446, 448-456.

82. The chimeric protein or protein complex or the Fc-based chimeric protein complex of claim 81 , wherein the chimeric protein or protein complex or Fc-based chimeric protein complex comprises a polypeptide having an amino acid sequence selected from one of SEQ ID NO: 439, 444, 453, and 455 and a polypeptide having an amino acid sequence selected from one of SEQ ID NO: 440-443 and 445-452.DBl / 162518159.2 134

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