Activatable il-10 polypeptides and methods of use thereof

Abstract

Provided herein are IL-10 prodrugs comprising IL-10, an IL-10 blocking element and a protease cleavable linker. The IL-10 prodrugs can optionally include a half-life extension element. Also provided herein are pharmaceutical compositions thereof, as well as nucleic acids, recombinant expression vectors, host cells for making such polypeptide prodrugs. Also disclosed are methods of using the IL-10 prodrugs in the treatment of diseases, conditions and disorders.

Description

Attorney Docket No.: 761146.192320 ACTIVATABLE IL-10 POLYPEPTIDES AND METHODS OF USE THEREOF

[0001] The present application claims the benefit of U.S. Provisional Application No.63 / 518,736 filed on August 10, 2023, the entire contents of which are incorporated herein by reference in their entirety. 1. BACKGROUND

[0002] Interleukin-10 (IL-10) is a secreted cytokine mainly produced by monocytes, macrophages, T cells (regulatory T cells and type-II T helper cells (Th2)) and mast cells. IL-10 is primarily regarded as an anti-inflammatory cytokine, but depending on the cell type, tissue and disease context, IL-10 can have pleiotropic actions. Brooks et al., (2010), Proc. Natl. Acad. Sci. U.S.A, 107(7), pp.3018-3023. IL-10 suppresses host inflammatory processes by myeloid and CD4+ T cells by enhancing regulatory T cell function or decreasing myeloid cell pro-inflammatory processes. Id. IL-10 secretion can also lead to increased activation and proliferation of a host of cell types, including CD8+ T cells and B cells. Id. Human and mouse IL-10 are noncovalently linked polypeptide homodimers. Zdanov et al., (1995), Structure, 3(6):591-601. IL-10 is recognized as being a key regulator of inflammation and genetic studies have shown that deficiency in IL-10 or IL-10 receptors is associated with mucosal inflammation and inflammatory bowel disease. (Sraiva M, et al. J. Exp. Med.2020; 217(1): e20190418). IL-10 is also reported to have a role in inflammatory disease, neurodegenerative disease, infection and cancer. Id. IL- 10 is also expressed in the tumor microenvironment of some cancers and is associated with poor prognosis. Id. IL-10 has been reported to have both tumor promoting and tumor inhibiting activities. Id.

[0003] The functional receptor for IL-10 (IL-10R) is a tetrameric transmembrane receptor composed of two unique α subunits (IL10RA) and two β subunits (IL10RB). The β subunits are shared with other cytokine receptors including IL-22R, the interferon lambda receptor and IL-26R. Monocytes and macrophages are the main target cell types harboring IL-10R, and binding of IL-10 to its receptor results in activation of the JAK / STAT signaling pathway. Signaling downstream of IL-10R binding to IL-10 leads to a decrease in phagocytosis, antigen presentation and an inhibition of pro-inflammatory cytokine release. Moreover, IL-10 signaling boosts the inhibitory and tolerogenic features of cells by, for example, increasing the release of interleukin-1 receptor antagonist, soluble TNFα receptor and IL-27.

[0004] IL-10 is an important anti-inflammatory cytokine that can suppress inflammation and, as such, represents a promising therapeutic agent for a variety of indications characterized by inflammation. However, the hoped-for clinical potential of IL-10 has not been achieved, due to undesired side effects, poor pharmacokinetics, stimulatory effects of IL-10 on antibody production by B cells and other effects in clinical studies. (Wang X., et al. Cold Spring Harbor Perspect Biol.2019:11(2): a028548). While clinical experience with IL-10 is limited, thus far there has been no clear clinical benefit observed. (Id.) 1 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0005] Inducible IL-12, IL-2, and interferon prodrugs have been described in International Publication Nos.: WO2019 / 222294, WO2019 / 222295, WO2019 / 222296, WO2021 / 097376.

[0006] A need exists for improved forms of IL-10 that can be used, for example, to effectively treat inflammatory and autoimmune diseases. 2. SUMMARY

[0007] The disclosure relates to inducible IL-10 prodrugs that contain at least one polypeptide chain, and can contain two or more polypeptides, if desired. The inducible IL-10 prodrug comprises an IL-10 polypeptide, an IL-10 blocking element, and a protease cleavable polypeptide linker. The IL-10 prodrug can further comprise a half-life extension element, if desired. The IL-10 polypeptide and the IL-10 blocking element can be operably linked by the protease-cleavable polypeptide linker.

[0008] Inducible IL-10 prodrugs of this disclosure have attenuated IL-10 receptor agonist activity and the circulating half-life is extended (relative to IL-10). The inducible IL-10 receptor agonist activity is attenuated through the blocking element. The half-life extension element can also contribute to attenuation, for example through steric effects. The blocking element is capable of blocking all or some of the receptor agonist activity of the IL-10 by noncovalently binding to the IL-10 and / or sterically blocking receptor binding. Upon cleavage of the protease cleavable linker a form of the IL-10 is released from the prodrug that is active (e.g., more active than the IL-10 polypeptide prodrug). Typically, the released IL-10 is at least 10 x more active than the IL-10 polypeptide prodrug. Preferably, the released IL-10 is at least 20 x, at least 30 x, at least 50 x, at least 100 x, at least 200 x, at least 300 x, at least 500 x, at least 1000 x, at least about 10,000X or more active than the inducible IL-10 prodrug. Accordingly, when the protease cleavable linker is a substrate for a protease whose activity is enriched, for example, in one or more locations within the body (e.g., inflammatory site, tumor microenvironment, organ), the prodrug can be preferentially cleaved by that protease and deliver active IL-10 to the location.

[0009] The form of IL-10 that is released upon cleavage of the inducible IL-10 prodrug typically has a short half-life, which is often substantially similar to the half-life of naturally occurring cytokine. Even though the half-life of the inducible IL-10 prodrug is extended, toxicity is reduced or eliminated because the agonist activity of the circulating inducible cytokine prodrug is attenuated and short half-life active IL-10 is targeted to the desired site of activity (e.g., inflammatory site, tumor microenvironment). Receptor activating activity of the inducible IL-10 prodrugs can be assessed by HEK-Blue reporter cell assay. IL-10 receptor activation can be determined using any standard in vitro receptor activation assay and equal amounts on a mole basis of the IL-10 polypeptide, released IL-10 and the inducible IL-10 prodrug. 2 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0010] The form of IL-10 that is released upon cleavage of the inducible IL-10 prodrug typically has a short half-life, which is often substantially similar to the half-life of naturally occurring cytokine. Even though the half-life of the inducible IL-10 prodrug may be extended, toxicity is reduced or eliminated because the agonist activity of the circulating inducible cytokine prodrug is attenuated and active IL-10 is targeted to or released at the desired site of activity (e.g., inflammatory site or tumor microenvironment).

[0011] The inducible IL-10 prodrug can be of the formulae:

[0012] [D1]-[L1]-[A]-[L1]-[D2],

[0013] [D1]-[L1]-[A]-[L2]-[A]-[L1]-[D2], or

[0014] [D1]-[L1]-[A]-[L1]-[D2]-[L1]-[A]-[L1]-[D3]. [A] is an interleukin 10 (IL-10) polypeptide, [D1], [D2] and [D3] are each, independently, a blocking element, and [L1] and [L2] are each independently a polypeptide linker. [L1] is a protease-cleavable linker and [L2] is optionally protease cleavable. The inducible IL-10 prodrugs disclosed herein can comprise an amino acid sequence from the group consisting of the following sequences SEQ ID NOS.: 59, 60, 62, 90, or 91 or an amino acid sequence that has at least about 80% identity to SEQ ID NOS.:59, 60, 62, 90, or 91.

[0015] The inducible IL-10 prodrug can optionally contain a half-life extension element [H]. The half- life extension element can be a serum albumin binding domain, an antigen binding polypeptide which binds human serum albumin, a serum albumin, human serum albumin, transferrin, or an immunoglobulin Fc, or fragment thereof. The inducible IL-10 prodrugs disclosed herein can comprise a half-life extension element [H], and have the formulae:

[0016] [A]-[L1]-[D]-[L2]-[H]; or

[0017] [A]-[L1]-[H]-[L2]-[D]; or

[0018] [H]-[L1]-[A]-[L4]-[D]; or

[0019] [D]-[L1]-[A]-[L4]-[H]; or

[0020] [H]-[L2]-[D]-[L1]-[A]; or

[0021] [D]-[L2]-[H]-[L1]-[A].

[0022] [L1], [L2], [L4] are independently a polypeptide linker. [L1] is a protease cleavable polypeptide linker, [L2] is a polypeptide linker that is optionally protease cleavable, and [L4] is a protease-cleavable polypeptide linker.

[0023] The inducible IL-10 prodrug can comprise one IL-10 polypeptide, two IL-10 polypeptides, or three IL-10 polypeptides. In some examples, the inducible IL-10 prodrug comprises one IL-10 polypeptide. In some examples, the inducible IL-10 prodrug comprises two IL-10 polypeptides. In some examples, the inducible IL-10 prodrug comprises three IL-10 polypeptides. 3 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0024] The IL-10 polypeptide can, be an IL-10 dimer. The IL-10 dimer can be covalently linked (e.g., protease cleavable linker, protease non-cleavable linker) or noncovalently bound (e.g., intramolecular binding, intermolecular binding). When the IL-10 polypeptide comprises two IL-10 polypeptides, the IL- 10 polypeptides can form a dimer by covalent or intramolecular binding. Alternatively, the IL-10 polypeptides can be optionally operably linked by a polypeptide linker.

[0025] When two or more IL-10 polypeptides are present, they can be operably linked by an optionally protease-cleavable linker. The IL-10 polypeptide [A] can have the formula: [A1]-[L3]-[A2], wherein [A1] and [A2] are each an IL-10 polypeptide and [L3] is a polypeptide linker that is optionally protease cleavable. [L3] can be cleaved by the same or a different protease as [L1] and / or [L2] and / or [L4]. [A1] and [A2] can have the same or different amino acid sequence.

[0026] The IL-10 blocking element [D] can sterically inhibit or block activation and / or binding of the IL- 10 polypeptide to its cognate receptor. The blocking element [D] can bind to the IL-10 polypeptide. The blocking element [D] can be human serum albumin, an antigen binding protein, or an antigen binding polypeptide which binds human serum albumin or a fragment thereof. The blocking element [D] can be a ligand binding domain or fragment of a cognate receptor for IL-10 (e.g., IL-10Ralpha or IL-10Rbeta), an antibody, or antibody fragment that binds to IL-10. When the IL-10 blocking element [D] is an antibody or antigen-binding fragment, it can be a single domain antibody, a Fab, or a scFv that binds the IL-10 polypeptide (e.g., binds a human IL-10 polypeptide). The blocking element [D] can optionally also extend the in vivo half-life. The inducible IL-10 prodrug can comprise two or more blocking elements (e.g., two blocking elements, three blocking elements). For example, the inducible IL-10 prodrug can comprise one blocking element, two blocking elements, three blocking elements, or more blocking elements.

[0027] The blocking element can comprise Blocking Element 1 (SEQ ID NO: 127 / SEQ ID NO: 168), Blocking Element 2 (SEQ ID NO: 128 / SEQ ID NO: 169), Blocking Element 3 (SEQ ID NO: 129 / SEQ ID NO: 170), Blocking Element 4 (SEQ ID NO: 130 / SEQ ID NO: 171), Blocking Element 5 (SEQ ID NO: 131 / SEQ ID NO: 172), Blocking Element 6 (SEQ ID NO: 132 / SEQ ID NO: 173), Blocking Element 7 (SEQ ID NO: 133 / SEQ ID NO: 174), Blocking Element 8 (SEQ ID NO: 134 / SEQ ID NO: 175), Blocking Element 9 (SEQ ID NO: 135 / SEQ ID NO: 176), Blocking Element 10 (SEQ ID NO: 136 / SEQ ID NO: 177), Blocking Element 10 (SEQ ID NO: 136 / SEQ ID NO: 177), Blocking Element 11 (SEQ ID NO: 137 / SEQ ID NO: 178), Blocking Element 12 (SEQ ID NO: 138 / SEQ ID NO: 179), Blocking Element 13 (SEQ ID NO: 139 / SEQ ID NO: 180), Blocking Element 14 (SEQ ID NO: 140 / SEQ ID NO: 181), Blocking Element 15 (SEQ ID NO: 141 / SEQ ID NO: 182), Blocking Element 16 (SEQ ID NO: 142 / SEQ ID NO: 183), Blocking Element 17 (SEQ ID NO: 143 / SEQ ID NO: 184), Blocking Element 18 (SEQ ID NO: 144 / SEQ ID NO: 185), Blocking Element 19 (SEQ ID NO: 145 / SEQ ID NO: 186), Blocking 4 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Element 20 (SEQ ID NO: 146 / SEQ ID NO: 187), Blocking Element 21 (SEQ ID NO: 147 / SEQ ID NO: 188), Blocking Element 22 (SEQ ID NO: 148 / SEQ ID NO: 189), Blocking Element 23 (SEQ ID NO: 149 / SEQ ID NO: 190), Blocking Element 24 (SEQ ID NO: 150 / SEQ ID NO: 191), Blocking Element 25 (SEQ ID NO: 151 / SEQ ID NO: 192), Blocking Element 26 (SEQ ID NO: 152 / SEQ ID NO: 193), Blocking Element 27 (SEQ ID NO: 153 / SEQ ID NO: 194), Blocking Element 28 (SEQ ID NO: 154 / SEQ ID NO: 500), Blocking Element 29 (SEQ ID NO: 155 / SEQ ID NO: 501), Blocking Element 30 (SEQ ID NO: 156 / SEQ ID NO: 502), Blocking Element 31 (SEQ ID NO: 157 / SEQ ID NO: 503), Blocking Element 32 (SEQ ID NO: 158 / SEQ ID NO: 504), Blocking Element 33 (SEQ ID NO: 159 / SEQ ID NO: 505), Blocking Element 34 (SEQ ID NO: 160 / SEQ ID NO: 506), Blocking Element 35 (SEQ ID NO: 161 / SEQ ID NO: 507), Blocking Element 36 (SEQ ID NO: 162 / SEQ ID NO: 507), Blocking Element 37 (SEQ ID NO: 163 / SEQ ID NO: 508), Blocking Element 38 (SEQ ID NO: 164 / SEQ ID NO: 509), Blocking Element 39 (SEQ ID NO: 165 / SEQ ID NO: 510), Blocking Element 40 (SEQ ID NO: 166 / SEQ ID NO: 512), Blocking Element 41 (SEQ ID NO: 167 / SEQ ID NO: 513), or any combinations of the foregoing.

[0028] The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 127 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 127, and a light chain constant region that comprises or consists of SEQ ID NO: 168 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 168. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 128 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 128, and a light chain constant region that comprises or consists of SEQ ID NO: 169 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 169. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 130 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 130, and a light chain constant region that comprises or consists of SEQ ID NO: 171 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 171. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 139 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 139, and a light chain constant region that comprises or consists of SEQ ID NO: 180 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 180. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 140 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 140, and a light chain constant region that comprises or consists of SEQ ID NO: 181 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 181.The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 144 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 144, and a light chain 5 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 constant region that comprises or consists of SEQ ID NO: 185 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 185. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 149 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 149, and a light chain constant region that comprises or consists of SEQ ID NO: 190 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 190. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 149 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 149, and a light chain constant region that comprises or consists of SEQ ID NO: 190 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 190. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 151 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 151, and a light chain constant region that comprises or consists of SEQ ID NO: 193 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 193. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 152 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 152, and a light chain constant region that comprises or consists of SEQ ID NO: 194 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 194. The blocking element can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 153 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 153, and a light chain constant region that comprises or consists of SEQ ID NO: 195 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 195.

[0029] The antibody or antigen-binding fragment can comprise an antibody light chain (VL + CL) comprising the amino acid sequence of any one of SEQ ID NOs: 127-167 and an antibody heavy chain Fab fragment (VH + CH1) comprising the amino acid sequence of any one of SEQ ID NOs: 168-194, and 500-513.

[0030] The inducible IL-10 prodrugs can contain two polypeptide chains. The inducible IL-10 prodrug can comprise comprises an IL-10 polypeptide, a half-life extension element, an IL-10 blocking element, and a protease cleavable linker. The IL-10 blocking element can be an antigen binding fragment of an antibody with separate components of at least an antigen-binding portion of an antibody light chain and at least an antigen-binding portion of a complementary antibody heavy chain. The inducible IL-10 can comprise a first polypeptide that can comprise an IL-10 polypeptide, at least an antigen binding portion of an antibody light chain or an antigen binding portion of an antibody heavy chain and a half-life extension element. The IL-10 polypeptide and the antigen binding portion of the antibody light chain or the antigen binding portion of the antibody heavy chain and / or half-life extension element can be operably linked by the protease cleavable linker. A second polypeptide can comprise at least an antigen binding portion of an 6 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 antibody heavy chain that is complementary to the light chain in the first polypeptide, or an antibody light chain that is complementary to the heavy chain in the first polypeptide. Together, the heavy and the light chains can comprise an IL-10 binding site.

[0031] The first polypeptide of the inducible IL-10 prodrug can comprise the formula of:

[0032] [A]-[L1]-[D]-[L2]-[H] or

[0033] [A]-[L1]-[H]-[L2]-[D] or

[0034] [H]-[L1]-[A]-[L4]-[D] or

[0035] [D]-[L1]-[A]-[L4]-[H] or

[0036] [H]-[L2]-[D]-[L1]-[A] or

[0037] [D]-[L2]-[H]-[L1]-[A]

[0038] [L1], [L2], and [L4] are each independently a polypeptide linker, wherein [L1] is a protease- cleavable polypeptide linker, [L2] is a polypeptide linker that is optionally protease cleavable, and [L4] is a protease-cleavable polypeptide linker. The antigen binding fragment of the antibody can be an antigen- binding portion of an antibody light chain or at least an antigen-binding portion of a complementary antibody heavy chain. The inducible IL-10 prodrug is an attenuated IL-10 receptor agonist, but upon (i) cleavage of the L1 protease-cleavable polypeptide linker, or (ii) cleavage of both L1 and L2 when L2 is a protease cleavable polypeptide linker, the IL-10 polypeptide-containing fragment of the inducible IL-10 prodrug that is produced is an IL-10 receptor agonist of comparable potency and half-life to the naturally occurring IL-10 polypeptide. The second polypeptide can comprise an antigen-binding portion of an antibody light chain or at least an antigen-binding portion of a complementary antibody heavy chain. When the antigen-binding portion of an antibody light chain is on the first polypeptide chain, the antigen- binding portion of a complementary antibody heavy chain is on the second polypeptide chain. When the antigen-binding portion of a complementary antibody heavy chain is on the first polypeptide chain, the antigen-binding portion of an antibody light chain is on the second polypeptide.

[0039] The inducible IL-10 prodrugs may contain one or more optionally protease-cleavable polypeptide linkers (e.g., [L1], [L2], [L3], [L4]) that are cleaved by a specific protease. The protease-cleavable linker can be cleaved by a protease selected from a kallikrein, a thrombin, a chymase, a carboxypeptidase, a cathepsin, an elastase, PR-3, a granzyme (e.g., granzyme M), a calpain, a matrix metalloproteinase (MMP), a fibroblast activation protein (FAP), an ADAM metalloproteinase, a plasminogen activator, a caspase, a tryptase, and a tumor cell protease. The cathepsin can be cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, cathepsin S, or cathepsin G. The matrix metalloprotease (MMP) can be MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, MMP19, or MMP20. 7 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0040] The protease cleavage site for each protease cleavable polypeptide linker [L] may be the same or different between each [L1], [L2], and [L3] and [L4]. The protease cleavable peptide linker(s) [L] can independently contain two or more cleavage sites for the same protease, two or more cleavage sites that are cleaved by different proteases or at least one of the protease-cleavable polypeptides comprises a cleavage site for two or more different proteases. The IL-10 polypeptide can freely dissociate from the IL- 10 blocking element [D], and the half-life extension element [H], if present, following cleavage of the protease-cleavable sequence(s). In some embodiments, the optionally protease cleavable linker is not protease cleavable.

[0041] The protease-cleavable polypeptide linkers (e.g., [L1], [L2], [L3], [L4]) of the inducible IL-10 prodrug can comprise an amino acid sequence selected from the group consisting of SEQ ID NOS.: 5, 9- 11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249, or 259 or have a sequence identity that has at least 90% sequence identity to SEQ ID NOS.: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112- 115, 195-201, 249, or 259. The protease-cleavable polypeptide linker can comprise SEQ ID NO.: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249, or 259, or a functional variant of SEQ ID NO.: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249, or 259. The protease- cleavable polypeptide linkers can have the amino sequence of SEQ ID NOS.: 37, 31, 195, or 249. The protease-cleavable polypeptide linkers can have the amino sequence of SEQ ID NO.: 37. The protease- cleavable polypeptide linkers can have the amino sequence of SEQ ID NO.: 195. The protease-cleavable polypeptide linkers can have the amino sequence of SEQ ID NO.: 198. The protease-cleavable polypeptide linkers can have the amino sequence of SEQ ID NO.: 38. The protease-cleavable polypeptide linkers can have the amino sequence of SEQ ID NO.: 39.

[0042] The inducible IL-10 prodrug can comprise any one of Compounds 1-93 and 97-100 as shown in Table 10. The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 68, 72-76, 81-87, 93-104, 116-119, 123-126, and 514-567, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NOs: 69, 70, 71, 568-621.

[0043] The inducible IL-10 prodrug can comprise Compound 1 (SEQ ID NO: 514 / SEQ ID NO: 568), Compound 2 (SEQ ID NO: 515 / SEQ ID NO: 569), Compound 3 (SEQ ID NO: 516 / SEQ ID NO: 570), Compound 4 (SEQ ID NO: 517 / SEQ ID NO: 571), Compound 5 (SEQ ID NO: 518 / SEQ ID NO: 572), Compound 6 (SEQ ID NO: 519 / SEQ ID NO: 573), Compound 7 (SEQ ID NO: 520 / SEQ ID NO: 574), Compound 8 (SEQ ID NO: 521 / SEQ ID NO: 575), Compound 9 (SEQ ID NO: 522 / SEQ ID NO: 576), Compound 10 (SEQ ID NO: 523 / SEQ ID NO: 577), Compound 11 (SEQ ID NO: 524 / SEQ ID NO: 578), Compound 12 (SEQ ID NO: 525 / SEQ ID NO: 579), Compound 13 (SEQ ID NO: 526 / SEQ ID NO: 580), 8 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Compound 14 (SEQ ID NO: 527 / SEQ ID NO: 581), Compound 15 (SEQ ID NO: 528 / SEQ ID NO: 582), Compound 16 (SEQ ID NO: 529 / SEQ ID NO: 583), Compound 17 (SEQ ID NO: 530 / SEQ ID NO: 584), Compound 18 (SEQ ID NO: 531 / SEQ ID NO: 585), Compound 19 (SEQ ID NO: 532 / SEQ ID NO: 586), Compound 20 (SEQ ID NO: 533 / SEQ ID NO: 587), Compound 21 (SEQ ID NO: 534 / SEQ ID NO: 588), Compound 22 (SEQ ID NO: 535 / SEQ ID NO: 589), Compound 22 (SEQ ID NO: 535 / SEQ ID NO: 589), Compound 23 (SEQ ID NO: 536 / SEQ ID NO: 590), Compound 24 (SEQ ID NO: 537 / SEQ ID NO: 591), Compound 25 (SEQ ID NO: 538 / SEQ ID NO: 592), Compound 26 (SEQ ID NO: 539 / SEQ ID NO: 593), Compound 27 (SEQ ID NO: 540 / SEQ ID NO: 594), Compound 28 (SEQ ID NO: 541 / SEQ ID NO: 595), Compound 29 (SEQ ID NO: 542 / SEQ ID NO: 596), Compound 30 (SEQ ID NO: 543 / SEQ ID NO: 597), Compound 31 (SEQ ID NO: 544 / SEQ ID NO: 598), Compound 32 (SEQ ID NO: 545 / SEQ ID NO: 599), Compound 33 (SEQ ID NO: 546 / SEQ ID NO: 600), Compound 34 (SEQ ID NO: 547 / SEQ ID NO: 601), Compound 35 (SEQ ID NO: 548 / SEQ ID NO: 602), Compound 36 (SEQ ID NO: 549 / SEQ ID NO: 603), Compound 37 (SEQ ID NO: 550 / SEQ ID NO: 604), Compound 38 (SEQ ID NO: 551 / SEQ ID NO: 605), Compound 39 (SEQ ID NO: 552 / SEQ ID NO: 606), Compound 40 (SEQ ID NO: 553 / SEQ ID NO: 607), Compound 41 (SEQ ID NO: 554 / SEQ ID NO: 608), Compound 42 (SEQ ID NO: 555 / SEQ ID NO: 609), Compound 43 (SEQ ID NO: 556 / SEQ ID NO: 610), Compound 44 (SEQ ID NO: 557 / SEQ ID NO: 611), Compound 45 (SEQ ID NO: 558 / SEQ ID NO: 612), Compound 46 (SEQ ID NO: 559 / SEQ ID NO: 613), Compound 47 (SEQ ID NO: 560 / SEQ ID NO: 614), Compound 48 (SEQ ID NO: 561 / SEQ ID NO: 615), Compound 49 (SEQ ID NO: 562 / SEQ ID NO: 616), Compound 50 (SEQ ID NO: 563 / SEQ ID NO: 617), Compound 51 (SEQ ID NO: 564 / SEQ ID NO: 618), Compound 52 (SEQ ID NO: 565 / SEQ ID NO: 619), Compound 53 (SEQ ID NO: 566 / SEQ ID NO: 620), or Compound 54 (SEQ ID NO: 567 / SEQ ID NO: 621).

[0044] The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 531, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 585. The inducible IL-10 prodrug can comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 532, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 586. The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 533, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 587. The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 534, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 588. The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the 9 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 amino acid sequence of SEQ ID NO: 535, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 589. The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 536, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 590. The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 540, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 594. The inducible IL-10 prodrug can comprise a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 542, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO: 596.

[0045] The inducible IL-10 prodrug can comprise an IL-10 polypeptide, a half-life extension element, an IL-10 blocking element and a protease cleavable linker. The IL-10 blocking element can be an antigen binding fragment of an antibody, wherein the antigen binding fragment comprises as separate components, at least an antigen-binding portion of an antibody light chain and at least an antigen-binding portion of a complementary antibody heavy chain, and the inducible IL-10 prodrug comprises: a first polypeptide comprising the IL-10 polypeptide, at least an antigen binding portion of an antibody light chain, and the half-life-extension element; wherein the IL-10 polypeptide and the antigen binding portion of the antibody light chain and / or half-life extension element are operably linked by the protease cleavable linker; and a second polypeptide comprising at least an antigen binding portion of an antibody heavy chain that is complementary to the light chain in the first polypeptide, together with said light chain forms an IL-10 binding site. The antigen binding portion of an antibody light chain can comprise any one of SEQ ID NOs: 168-194 and 500-513 and the protease-cleavable peptide linker can comprises any one of SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249 or 259. The antigen binding portion of an antibody heavy chain comprises any one of SEQ ID NOs: 514-567.

[0046] The disclosure also relates to a nucleic acid composition comprising one or more nucleic acid sequences encoding an inducible IL-10 prodrug disclosed herein. For example, the nucleic acid compositions comprising one or more nucleic acid sequences encoding an inducible IL-10 prodrug can comprise at least one of each of: an IL-10 polypeptide [A], an IL-10 blocking element [D], and a protease-cleavable polypeptide linker [L]. The IL-10 polypeptide, and the IL-10 blocking element are operably linked by the protease-cleavable polypeptide linker and the inducible IL-10 has attenuated IL-10 receptor activating activity. The IL-10 receptor activating activity of the inducible IL-10 prodrug is at least about 10 x less than the IL-10 receptor activating activity of the polypeptide that contains the IL-10 polypeptide that is produced by cleavage of the protease-cleavable linker. For example, the nucleic acid 10 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 composition comprising one or more nucleic acid sequences encoding an inducible IL-10 prodrug can comprise an IL-10 polypeptide, a half-life extension element, an IL-10 blocking element, and a protease cleavable linker. The IL-10 blocking element can be an antigen binding fragment of an antibody. The antigen binding fragment can comprise as separate components, at least an antigen-binding portion of an antibody light chain and at least an antigen-binding portion of a complementary antibody heavy chain. The inducible IL-10 prodrugs disclosed herein can comprise a first polypeptide comprising the IL-10 polypeptide, at least an antigen binding portion of an antibody light chain or an antigen binding portion of an antibody heavy chain, and the half-life-extension element. The IL-10 polypeptide and the antigen binding portion of the antibody light chain or the antigen binding portion of the antibody heavy chain and / or half-life extension element are operably linked by the protease cleavable linker; and a second polypeptide comprising at least an antigen binding portion of an antibody heavy chain that is complementary to the light chain in the first polypeptide, or an antibody light chain that is complementary to the heavy chain in the first polypeptide and together with said light chain forms an IL-10 binding site.

[0047] The nucleic acid composition encoding inducible IL-10 prodrug(s) can be a circular vector, DNA, RNA, or an expression vector comprising a circular vector, DNA, or RNA. Additionally, the disclosure relates to an isolated host cell comprising an expression vector encoding for a nucleic acid composition of an inducible IL-10 prodrug.

[0048] The disclosure also relates to a pharmaceutical composition(s) of the inducible IL-10 prodrug and methods of making pharmaceutical compositions of the inducible IL-10 prodrug. Methods of making such a pharmaceutical composition can comprise culturing isolated host cells (comprising an expression vector of the inducible IL-10 prodrug) under suitable conditions for expression and collection of desired inducible IL-10 prodrugs as polypeptides. The methods disclosed herein can further comprise isolating the inducible IL-10 prodrug. The pharmaceutical composition can comprise an effective amount of the inducible IL-10 prodrug or nucleic acid encoding for the inducible IL-10 prodrug.

[0049] The disclosure also relates to methods for treating inflammatory conditions that comprise administering to a subject in need thereof an effective amount of the inducible IL-10 prodrug, a nucleic acid encoding the inducible IL-10 prodrug, an expression vector for the inducible IL-10 prodrug, or a pharmaceutical composition comprising an effective amount of any of these. 3. BRIEF DESCRIPTION OF DRAWINGS

[0050] The drawings are not necessarily to scale or exhaustive. Instead, the emphasis is generally placed upon illustrating the principles of the inventions described herein. The accompanying drawings, which 11 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 constitute a part of the specification, illustrate several embodiments consistent with the disclosure and, together with the description, serve to explain the principles of the disclosure. In the drawings:

[0051] FIGs.1A-1E are schematic illustrations depicting various inducible IL-10 prodrugs. The rectangles included between the elements indicates that the linker is a protease cleavable linker. When a rectangle is not present between the elements the linker is optionally protease cleavable.

[0052] FIGs.2A, 2C, 2E are graphs showing activity of IL-10 prodrugs in an HEK-Blue IL-10 reporter assay in the presence of HSA and FIGs.2B, 2D, and 2F are images of SDS-PAGE gels showing the results of protein cleavage assays. FIG.2A depicts reporter activation in a comparison of WW0857 inducible IL-10 prodrug to human IL-10 (control). FIG.2C depicts reporter activation in a comparison of WW0858 inducible IL-10 prodrug to human IL-10 (control). FIG.2E depicts reporter activation in comparison of WW0860 inducible IL-10 prodrug to human IL-10 (control). Squares depict activity of the uncleaved inducible IL-10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity of the control human IL-10. EC50 values for each are shown in the table (N.D. = not determined). Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible IL-10 prodrugs are active and inducible. FIGs.2B, 2D, and 2F show the results of protein cleavage assays. WW0857 (FIG.2B), WW0858 (FIG.2D), WW0860 (FIG.2F) were run on an SDS- PAGE gel. +CTSL refers to Cathepsin L run in non-reduced conditions, NR refers to non-reduced conditions, and R refers to reduced conditions. The inducible IL-10 prodrugs were run in both cleaved and uncleaved form.

[0053] FIG.3A is a graph showing activity of half-life extended IL-10 n a HEK-Blue IL-10 reporter assay in the presence of HSA. Triangles depict the activity of the WW00976 inducible IL-10 prodrug. Circles depict activity of the control human IL-10 (WW0882). EC50 values for each are shown in the table (N.D. = not determined). FIG.3B is an image of an SDS-PAGE gel showing the results of the protein cleavage assay. NR refers to non-reduced conditions, and R refers to reduced conditions.

[0054] FIGs.4A, 4C, and 4E are graphs showing activity of inducible IL-10 prodrugs containing an IL- 10 monomer in an HEK-Blue IL-10 reporter assay in the presence of HSA and FIGs.4B, 4D, and 4F are images of SDS-PAGE gels showing the results of protein cleavage assays. FIG.4A depicts reporter activation in a comparison of WW00909 / WW0911 inducible IL-10 prodrug to human IL-10 (control). FIG.4C depicts reporter activation in a comparison of WW0915 / WW0911 inducible IL-10 prodrug to human IL-10 (control). FIG.4E depicts reporter activation in a comparison of WW0916 / WW0911 inducible IL-10 prodrug to human IL-10 (control). Squares depict activity of the uncleaved inducible IL- 10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity 12 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 of the control human IL-10. EC50 values for each are shown in the table. Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible IL-10 prodrugs are active and inducible. FIGs.4B, 4D, and 4F show the results of protein cleavage assays. WW00909 / WW0911 (FIG.2B), WW0915 / WW0911 (FIG.2D), WW0916 / WW0911 (FIG.2F) were run on an SDS-PAGE gel. NR refers to non-reduced conditions, and R refers to reduced conditions. The inducible IL-10 prodrugs were run in both cleaved and uncleaved form.

[0055] FIGs.5A, 5C, and 5E are graphs showing activity of inducible IL-10 prodrugs containing a IL- 10 dimer in an HEK-Blue IL-10 reporter assay in the presence of HSA and FIGs.5B, 5D, and 5F are images of SDS-PAGE gels showing the results of protein cleavage assays. FIG.5A depicts reporter activation in a comparison of WW0915 / WW0912 inducible IL-10 prodrug to human IL-10 (control). FIG.5C depicts reporter activation in a comparison of WW0915 / WW0914 inducible IL-10 prodrug to human IL-10 (control). FIG.5E depicts reporter activation in a comparison of WW0916 / WW0912 inducible IL-10 prodrug to human IL-10 (control). Squares depict activity of the uncleaved inducible IL- 10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity of the control human IL-10. EC50 values for each are shown in the table. Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible IL-10 prodrugs are active and inducible. FIGs.5B, 5D, and 5F show the results of protein cleavage assays. WW0915 / WW0912 (FIG.2B), WW0915 / WW0914 (FIG. 2D), WW0916 / WW0912 (FIG.2F) were run on an SDS-PAGE gel. The inducible IL-10 prodrugs were run in both cleaved and uncleaved form.

[0056] FIGs.6A, 6C, and 6E are graphs showing activity of inducible IL-10 prodrugs containing an Fc domain for half-life extension and blockage in an HEK-Blue IL-10 reporter assay in the presence of HSA and FIGs.6B, 6D, and 6F are images of SDS-PAGE gels showing the results of protein cleavage assays. FIG.6A depicts reporter activation in a comparison of WW0918, a half-life extended IL-10, to human IL-10 (control). FIG.6C depicts reporter activation in a comparison of WW0919 inducible IL-10 prodrug to human IL-10 (control). FIG.6E depicts reporter activation in a comparison of WW0920, a half-life extended IL-10, to human IL-10 (control). Squares depict activity of the uncleaved inducible IL-10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity of the control human IL-10. EC50 values for each are shown in the table. Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible Fc fusions are active. FIGs.6B, 6D, and 6F show the results 13 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 of protein cleavage assays. WW0918 (FIG.2B), WW0919 (FIG.2D), WW0920 (FIG.2F) were run on an SDS-PAGE gel. The inducible IL-10 prodrugs were run in both cleaved and uncleaved form.

[0057] FIGs.7A, 7C, 7E, 7G, 7I, 7K, 7M, 7O, 7Q, 7S, 7U, 7W, 7Y, 7ZA, 7ZC, 7ZE, 7ZG, and 7ZI are graphs showing activity of inducible IL-10 prodrugs in an HEK-Blue IL-10 reporter assay in the presence of HSA. Squares depict activity of the uncleaved inducible IL-10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity of the control human IL-10. EC50 values for each are shown in the table (N.D. = not determined). Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible IL-10 prodrugs are active and inducible. FIGs.7B, 7D, 7F, 7H, 7J, 7L, 7N, 7P, 7R, 7T, 7V, 7X, 7Z, 7ZB, 7ZD, 7ZF, 7ZH, and 7ZJ are images of SDS-PAGE gels showing the results of protein cleavage assays.

[0058] FIGs.8A, 8C, 8E, 8G are graphs showing activity of inducible IL-10 prodrugs containing a protease cleavable linker comprising SEQ ID NO.: 195 (FIGs.8A-8B), SEQ ID NO: 259 (FIGs.8C-8D), SEQ ID NO: 249 (FIGs.8E-8F), or SEQ IN NO: 14 (FIGs.8G-8H) in a protease cleavable linker in an HEK-Blue IL-10 reporter assay in the presence of HSA. Squares depict activity of the uncleaved inducible IL-10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity of the control human IL-10. EC50 values for each are shown in the table (N.D. = not determined). Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible IL-10 prodrugs are active and inducible. FIGs.8B, 8D, 8F, and 8H are images of SDS-PAGE gels showing the results of protein cleavage assays.

[0059] FIGs.9A, 9C, and 9E are graphs showing activity of inducible IL-10 prodrugs containing a protease cleavable linker comprising SEQ ID NO: 37 (FIGs.9A-9B), SEQ ID NO: 38 (FIGs.9C-9D) or SEQ ID NO: 41 (FIGs.9E-9F) in an HEK-Blue IL-10 reporter assay in the presence of HSA. Squares depict activity of the uncleaved inducible IL-10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity of the control human IL-10. EC50 values for each are shown in the table (N.D. = not determined). Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible IL-10 prodrugs are active and inducible. FIGS.9B, 9D, and 8F are images of SDS-PAGE gels showing the results of protein cleavage assays.

[0060] FIGs.10A, 10C, 10E are graphs showing activity of inducible IL-10 prodrugs containing a protease cleavable linker comprising SEQ IN NO: 40 (FIG.10A), SEQ ID NO: 39 (FIG.10C), or SEQ ID NO: 43 (FIG.10E) in an HEK-Blue IL-10 reporter assay in the presence of HSA. Squares depict activity 14 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 of the uncleaved inducible IL-10 prodrug and triangles depict the activity of the cleaved inducible IL-10 prodrug. Circles depict activity of the control human IL-10. EC50 values for each are shown in the table (N.D. = not determined). Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue® (InvivoGen). Results confirm that inducible IL-10 prodrugs are active and inducible. FIGs.10B, 10D, 10F are images of an SDS-PAGE gel showing the results of protein cleavage assays.

[0061] FIGs.11A-11D are graphs showing the activity of inducible IL-10 prodrugs in a primary cell assay. PBMCs were cultured with IL-10 prodrugs (uncleaved or cleaved) and pStat3 protein levels (%pStat3) in CD8+ cells were measured by flow cytometry. Circles depict activity of the uncleaved inducible IL-10 prodrug (uncleaved), and squares depict the activity of the cleaved inducible IL-10 prodrugs (cleaved). Triangles depict activity of an IL-10 forced dimer control (WW00882). Unstimulated cells depicted by diamonds and single dose controls of IL-10 forced dimer control (WW00882) depicted by asterisk. EC50 values for each are shown in the adjacent tables (N.D. = not determined).

[0062] FIGs.12A-12D are graphs showing the activity of inducible IL-10 prodrugs in a primary cell assay in which pStat3 protein levels (%pStat3) in CD4+ T cells were measured. Circles depict activity of the uncleaved IL-10 prodrug (uncleaved), and squares depict the activity of the cleaved inducible IL-10 prodrugs (cleaved). Triangles depict activity of an IL-10 forced dimer control (WW00882). Unstimulated cells depicted by diamonds and single dose controls of IL-10 forced dimer control (WW00882) depicted by asterisk. EC50 values for each are shown in the adjacent tables (N.D. = not determined).

[0063] FIGs.13A-13C are images of capillary western gels showing cleavage of IL-10 prodrugs following ex vivo incubation with lung dissociated tissue from naïve or asthmatic mice with the IL-10 prodrugs. Uncleaved inducible IL-10 prodrugs detected at 65-75 kDa and fully cleaved released IL-10 was detected at 42 kDA. FIGs.13D-13E are graphs depicting the quantification of aforementioned cleavage by plotting the percentage of fully cleaved inducible IL-10 prodrugs following ex vivo incubation with lung dissociated tissue from naïve or asthmatic mice with the IL-10 prodrugs. The results show that inducible IL-10 prodrugs are preferentially processed by inflamed murine lung tissue.

[0064] FIGs.14A and 14B, and 14D are flow cytometry plots with gates showing pStat3 expression in CD8+ T cells after incubation with supernatants from ex vivo incubation of murine lung cells and IL-10 prodrug. FIGs.14C and 14E are graphs depicting the fold change over WW01000 (control not expected to be cleaved in this assay) in percentage of pStat3+ for CD8+T cells for inducible IL-10 prodrugs comprising different linkers.

[0065] FIGs.15A-15B are images of capillary western gels showing cleavage of IL-10 prodrugs following ex vivo incubation with colon dissociated tissue from naïve or DSS (dextran sodium sulfate) 15 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 treated mice with the IL-10 prodrugs. Uncleaved inducible IL-10 prodrugs detected at 65-75 kDa and fully cleaved released IL-10 was detected at 42 kDA. FIGs.15C is a graph depicting the quantification of aforementioned cleavage by plotting the percentage of fully cleaved inducible IL-10 prodrugs following ex vivo incubation with colon dissociated tissue from naïve or DSS treated mice with the IL-10 prodrugs. FIGs.15D is a graph depicting fold change in activity of cleaved IL-10 prodrugs over WW01006 / WW00911 (noncleavable control) as measured by SEPA activity in the HEK-Blue IL-10 Reporter assay. These results show that inducible IL-10 prodrugs are preferentially processed by inflamed murine colon tissue.

[0066] FIGs.16A-16B are images of capillary western gels showing cleavage of IL-10 prodrugs following ex vivo incubation with healthy human colon cells or dissociated colon tissue from ulcerative colitis or Crohn’s patients with the IL-10 prodrugs. Uncleaved inducible IL-10 prodrugs detected at 65-75 kDa and fully cleaved released IL-10 was detected at 42 kDA. FIGs.16C is a graph depicting the quantification of aforementioned cleavage by plotting the percentage of fully cleaved inducible IL-10 prodrugs following ex vivo incubation with healthy human colon cells or dissociated colon tissue from ulcerative colitis or Crohn’s patients with the IL-10 prodrugs. FIGs.16D is a graph depicting fold change in activity of cleaved IL-10 prodrugs over WW01006 / WW00911 (non-cleavable control) as measured by SEPA activity in the HEK-Blue IL-10 Reporter assay. These results show that inducible IL-10 prodrugs are preferentially processed by inflamed human colon tissue compared to healthy colon cells.

[0067] FIG.17 is a graph of percent weight change in a study using the DSS model of colitis. C57BL / 6 mice were given 4% DSS in drinking water on days 1-6 and returned to normal drinking water for days 6- 24. Mice monitored daily for body weight loss and overall body condition. Percent weight change calculated for each animal relative to weight on day 1 prior to dosing and DSS addition in water. Circles depict weight change of naïve animals (no DSS). Squares depict weight change of vehicle treated mice (with DSS). Triangles depict weigh change of mice treated with anti-mouse IL-12p40 (with DSS). Diamonds depict weight change of mice treated with WW0976 (with DSS). Statistical analysis determined by 2-way ANOVA Dunnett’s multiple comparison test comparing treatment groups to vehicle group. Asterisks represent statistical significance between WW0976 treated and vehicle group.

[0068] FIGs 18A-18C are graphs of percent weight change in a study using the DSS model of colitis. FIG.18A depicts percent weight change over 24 days in vehicle. FIG.18B depicts percent weight change over 24 days in anti-p40. FIG.18C depicts percent weight change over 24 days in WW0976.

[0069] FIGs.19A and 19B are graphs depicting the quantification of cleavage by plotting the percentage of fully cleaved inducible IL-10 prodrugs having different cleavable linker substrates following ex vivo incubation with naïve colon samples and colon samples from dextran sulfate sodium 16 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 (DSS)-induced colitis in mice (FIG.19A) or naïve lung samples and samples from ovalbumin (OVA) acute allergic asthma model in mice (FIG.19B). These results show that unique substrates are required for different inflammatory diseases.

[0070] FIGs.20A-20E are graphs depicting exposure of inducible IL-10 prodrugs in tissue from a DSS- induced colitis mouse model. FIG.20A shows the percentage of weight change in the DSS-induced colitis mouse model over time administered with inducible IL-10 prodrugs. FIG.20B is a graph showing the concentration (pg / ml) of inducible IL-10 prodrug in serum from DSS-induced colitis mice and healthy mice. The results show similar systemic exposure of inducible IL-10 prodrugs in naïve and diseased animals. FIG.20C is a graph showing the amount of inducible IL-10 prodrug (pg per µg tissue) over time in colon tissue lysate from DSS-induced colitis mice and healthy mice. The graph shows increased exposure of inducible IL-10 in colon of diseased animals. FIG.20D is a graph showing the amount of free cytokine (pg per µg tissue) over time in colon tissue lysate from DSS-induced colitis mice and healthy mice. The graph shows release of payload cytokine in colon of diseased animals only by cleavable inducible IL-10 prodrug. Free cytokine was not observed for the non-cleavable inducible IL-10 prodrug or naïve samples. FIG.20E is a graph showing the concentration (pg / ml) of free human IL-10 over time in serum from DSS-induced colitis mice and healthy mice treated with non-cleavable inducible IL-10 prodrug or an inducible IL-10 prodrug.

[0071] FIGs.21A-21E show the efficacy of inducible IL-10 prodrugs in DSS induced colitis model. FIG.21A is a graph showing the percentage weight change over time in IL-10 knock-out Balb / c mice (a DSS induced colitis model) administered with naïve, vehicle plus 4% DSS, half-life extended IL-10 plus 4% DSS, and POC inducible IL-10 prodrug plus 4% DSS. FIG.21B is a microscopy image of tissue administered vehicle and inducible IL-10 prodrug. FIG.21C is a graph showing colon weight in mice administered with naïve, vehicle plus 4% DSS, half-life extended IL-10 plus 4% DSS, and POC inducible IL-10 prodrug plus 4% DSS. The graphs show reduced colon weight with administration of inducible IL- 10 prodrug. FIG.21D is a graph showing the level of TNFalpha expression in the colon in mice administered with naïve, vehicle plus 4% DSS, half-life extended IL-10 plus 4% DSS, and POC inducible IL-10 prodrug plus 4% DSS. The results show reduced inflammatory cytokines with administration of inducible IL-10 prodrug. FIG.21E is a graph showing the level of TNFalpha expression in serum administered with naïve, vehicle plus 4% DSS, half-life extended IL-10 plus 4% DSS, and POC inducible IL-10 prodrug plus 4% DSS.

[0072] FIGs.22A-22E are graphs showing body weight (FIG.22A), stool score (FIG.22B), disease activity index (FIG.22C), colon weight (FIG.22D), and colon weight:length ratio, and (FIG.22E) over days post cell transfer in the adoptive cell therapy (ACT) mouse model of colitis. 17 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0073] FIGs.23A and 23B are graphs showing percent in weight change over days in ACT colitis mice administered with vehicle, anti-IL-12, and inducible IL-10 prodrugs, and mice with no colitis. The graphs show that mice administered with inducible IL-10 prodrugs do not lose weight.

[0074] FIGs.24A and 24B are graphs showing stool score endpoint (FIG.24A) and disease activity index (FIG.24B) in mice ACT colitis mice administered with vehicle, anti-IL-12, and inducible IL-10 prodrugs, and mice with no colitis. The graphs show that inducible IL-10 prodrugs produce a significant effect on endpoint stool and disease activity index (DAI) scores and reduced the stool and DAI scores over time.

[0075] FIGs.25A-25C are graphs showing colon weight (FIG.25A), colon length (FIG.25B), and colon weight / length ratio (FIG.25C) in mice administered with vehicle, anti-IL-12, and inducible IL-10 prodrugs, and mice with no colitis.

[0076] FIGs.26A-26E are graphs showing expression of cytokines (IFNγ, IL-1β, TNF-alpha, KC / GRO, IL-17A in mice administered with vehicle, anti-IL-12, and inducible IL-10 prodrugs, and mice with no colitis. The graphs show inflammatory cytokines were reduced in colons of mice treated with inducible IL-10 prodrugs.

[0077] FIG.27 are microscopy images from mice with no colitis, vehicle, and inducible IL-10 prodrugs. The images show that there are no signs of colitis after treatment with inducible IL-10 prodrugs.

[0078] FIGs.28A-28B are graphs showing cleavage of IBD specific linkers by colon samples from human IBD patient samples (FIG.28A) and healthy human primary cells (FIG.28B). The graphs show that the IBD linkers are preferentially cleaved by human IBD samples and are stable when incubated with human primary cells. 4. DETAILED DESCRIPTION A. Inducible IL-10 Prodrugs

[0079] The inducible IL-10 prodrug can comprise a single polypeptide chain, or two or more polypeptide chains. Typically, when the IL-10 prodrug is a single polypeptide chain, the single polypeptide chain comprises at least one IL-10 polypeptide (e.g., a single IL-10 monomer polypeptide, two IL-10 monomer polypeptides, or three or more IL-10 monomer polypeptides) or a mutein thereof [A], a blocking element [D], a protease cleavable linker [L], and optionally a half-life extension element [H]. In embodiments where the optional half-life extension element is absent, it is preferred that the blocking element can also function as half-life extending element as described herein, e.g., and antigen binding fragment of an antibody that binds HSA and sterically inhibits binding of the IL-10 in the prodrug to the IL-10 receptor. The IL-10 polypeptide [A] can be operably linked to the blocking element, the half-life extension element (when present), or both the blocking element and the half-life extension element (when present) by a 18 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 protease cleavable linker. The single polypeptide chain can comprise about one, about two, about three, about four, about five, or more IL-10 polypeptides (e.g., IL-10 polypeptide monomers) or muteins thereof. Typically, the single polypeptide chain comprises one IL-10 polypeptide or two IL-10 polypeptides. The IL-10 polypeptide can be located at any desired position in the single polypeptide chain. When one IL-10 polypeptide is present, the IL-10 polypeptide monomer can non-covalently dimerize with a free IL-10 polypeptide. When two IL-10 polypeptides are present, the IL-10 polypeptides can form a dimer. The IL-10 polypeptides can form a dimer by covalent or intermolecular binding and / or optionally be operably linked by a polypeptide linker, which can be cleavable or not cleavable. For example, the IL-10 prodrug can include an IL-10 polypeptide [A] that contains two IL-10 monomers and can, for example, have the formula: [A1]-[L3]-[A2], wherein [A1] and [A2] are each an IL-10 polypeptide and [L3] is a polypeptide linker that is optionally protease cleavable. [A1] and [A2] can have the same or different amino acid sequence.

[0080] The single polypeptide can comprise two or more blocking elements that also function as half-life extension elements (e.g., an antibody fragment that binds HSA). When two or more of such blocking elements are present in the IL-10 prodrug, they can block all or some of the receptor agonist activity of IL-10 and also extend serum half-life. When two or more such a blocking element are present in and IL- 10 prodrug, a separate half-life extension element or a separate blocking element are optional and are typically not present. The one or more IL-10 polypeptides are operably linked by a protease cleavable linker to such a blocking element. For example, the polypeptide can have the formula:

[0081] [D1]-[L1]-[A]-[L1]-[D2] (VII);

[0082] [D1]-[L1]-[A]-[L2]-[A]-[L1]-[ D2] (VIII); or

[0083] [D1]-[L1]-[A]-[L1]-[ D2]-[L1]-[A]-[L1]-[ D3] (IX), wherein: [A] is an IL-10 polypeptide. The IL-10 polypeptide can be one or more copies of IL-10, [D1], [D2], and [D3] are independently a blocking element, [L1] is a protease-cleavable polypeptide linker, [L2] is a polypeptide linker that is optionally protease-cleavable. [L1] and [L2] can have the same or different amino acid sequence and / or protease-cleavage site (when L2 is protease-cleavable) as desired. [D1], [D2], or [D3] can also function as a half-life extension element.

[0084] The IL-10 polypeptide and the blocking element and / or the half-life extension element (when present) can be operably linked by the protease-cleavable polypeptide. For example, the polypeptide can be any of Formulas (I)-(VI).

[0085] [A]-[L1]-[H]-[L2]-[D] (I);

[0086] [D]-[L2]-[H]-[L1]-[A] (II);

[0087] [A]-[L1]-[D]-[L2]-[H] (III); 19 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0088] [H]-[L2]-[D]-[L1]-[A] (IV);

[0089] [H]-[L1]-[A]-[L4]-[D] (V); and

[0090] [D]-[L1]-[A]-[L4]-[H] (VI); wherein [A] is an IL-10 polypeptide (e.g., a single IL-10 monomer polypeptide, two IL-10 monomer polypeptides, or three or more IL-10 monomer polypeptides), [D] is an IL-10 blocking element (e.g., IL- 10R (i.e., IL10Ralpha or IL10Rbeta) or an antibody or antigen-binding fragment), [H] is a half-life extension element, [L1] is a protease-cleavable polypeptide linker, [L2] is a polypeptide linker that is optionally protease-cleavable, and [L4] is a protease-cleavable polypeptide linker. [L1] and [L2], or [L1] and [L4], can have the same or different amino acid sequence and / or protease-cleavage site (when L4 is protease-cleavable) as desired. [H] can also optionally provide blocking.

[0091] The single polypeptide IL-10 prodrugs disclosed herein can comprise or consist of the amino acid sequence of SEQ ID NO: 59, 60, and 62. For example, the inducible IL-10 prodrug can comprise the amino acid sequence of SEQ ID NO: 59. For example, the inducible IL-10 prodrug can comprise the amino acid sequence of SEQ ID NO: 60. For example, the inducible IL-10 prodrug can comprise the amino acid sequence of SEQ ID NO: 62. In some instances, the inducible IL-10 prodrug comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NOs: 59, 60, or 62.

[0092] The inducible IL-10 prodrug can comprise two or more polypeptide chains. When the IL-10 prodrug comprises two or more polypeptide chain. Such inducible IL-10 prodrugs comprises at least one IL-10 polypeptide (e.g., a single IL-10 monomer polypeptide, two IL-10 monomer polypeptides, or three or more IL-10 monomer polypeptides that can form a dimer by covalent or intermolecular binding and / or optionally be operably linked by a polypeptide linker, that can be cleavable or not cleavable) or a mutein thereof [A], a blocking element [D], a protease cleavable linker [L], and optionally a half-life extension element [H], which can be present on the same polypeptide chain or different polypeptide chains. For example, a first IL-10 polypeptide monomer can be present on a first polypeptide chain and a second IL- 10 polypeptide monomer can be present on a second polypeptide chain. Similarly, the blocking element and half-life extension element (when present) can contain two or more components that are present on the same polypeptide chain or on different polypeptide chains. Illustrative of this, and as disclosed and exemplified herein, components of the blocking element can be present on separate polypeptide chains. For example, a first polypeptide chain can include an antibody light chain (VL+CL) or light chain variable domain (VL) and a second polypeptide can include an antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) that is complementary to the VL+ CL or VL on the first 20 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 polypeptide. In such situations, these components can associate in the peptide complex to form an antigen-binding site, such as a Fab that binds IL-10 and attenuates IL-10 activity.

[0093] For example, the inducible IL-10 prodrug can comprise a first polypeptide of any of Formulas X- XVI.

[0094] Formula X: [D]-[L1]-[A]-[L4]-[H];

[0095] Formula XI: [H]-[L1]-[A]-[L4]-[D];

[0096] Formula XII: [A]-[L1]-[H]-[L2]-[D];

[0097] Formula XIII: [D]-[L2]-[H]-[L1]-[A];

[0098] Formula XIV: [A]-[L1]-[D]-[L2]-[H]; or

[0099] Formula XVI: [H]-[L2]-[D]-[L1]-[A], wherein: [A] is a IL-10 polypeptide. The IL-10 polypeptide can be one or IL-10 polypeptides (i.e., one IL-10 polypeptide, two IL-10 polypeptides, three IL-10 polypeptides or more), [D] is an anti-IL-10 antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) or an anti-IL-10 light chain Fab fragment (VL +CL) or light chain variable domain (VL), [H] is a half-life extension element, [L1] is a protease-cleavable polypeptide linker, and [L4] is a protease-cleavable linker, and [L2] is a polypeptide linker that is optionally protease-cleavable. [L1] and [L4] can have the same or different amino acid sequence and or protease-cleavage site, as desired. The inducible IL-10 prodrug can also have a second polypeptide comprising an antibody light chain (VL+CL) or light chain variable domain (VL) that is complementary to the VH + CH1 or VH on the first polypeptide or an antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) that is complementary to the VL + VC or VL on the first polypeptide. When the first polypeptide comprises a VH + CH1 or a VH, the first polypeptide can associate with a complementary anti-IL-10 VL+ CL or VL to form an inducible IL-10 prodrug that contains an antibody fragment (e.g., Fab) that binds IL-10. When the first polypeptide comprises a VL + VC or VL, the first polypeptide can associate with a complementary anti-IL-10 antibody VH + CH1 or VH to form an inducible IL-10 prodrug that contains an antibody fragment (e.g., Fab) that binds IL-10.

[0100] In embodiments, the inducible IL-10 prodrug can comprise a first polypeptide, such as a polypeptide of any of Formulas X – XVI, which is bonded covalently or non-covalently to a second polypeptide chain. The second polypeptide chain can contain an antibody VL-CL that comprises or consists of the amino acid sequence of SEQ ID NO: 69, SEQ ID NO; 70 or SEQ ID NO: 71. Such a second polypeptide can bond with a complimentary VH-CH1 polypeptide contained within the IL-10 prodrug, e.g., as contained within SEQ ID NOs.68, 69, 73-76, 81-87, 92-104 or 622.

[0101] The inducible IL-10 prodrug disclosed herein can comprise a first polypeptide chain that includes an IL-10 polypeptide and a IL-10 antibody heavy chain Fab fragment (VH + CH1) or heavy chain 21 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 variable domain (VH) that are linked through a protease cleavable linker, and a second polypeptide chain that includes a half-life extension element and an IL-10 antibody light chain (VL+CL) or light chain variable domain (VL) that is complementary to the VH + CH1 or VH on the first polypeptide that are linked through an optionally protease cleavable linker.

[0102] The inducible IL-10 prodrug disclosed herein can comprise a first polypeptide chain that includes a IL-10 polypeptide and an antibody light chain (VL+CL) or light chain variable domain (VL) that are linked through a protease cleavable linker, and a second polypeptide chain that includes a half-life extension element and a IL-10 antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) that is complementary to the VL+CL or VL of the first polypeptide chain and which are linked through an optional protease cleavable linker.

[0103] In embodiments, at least one of an IL-10 polypeptide monomer, the optional half-life extension element, the blocking element, or a component of the half-life extension element or blocking element is on a separate polypeptide. For example, the first polypeptide can comprise IL-10 and either an antibody light chain (VL + CL) or light chain variable domain (VL), or an antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) that is operably linked to IL-10 through a protease cleavable linker. The second polypeptide can comprise the half-life extension element and either an antibody light chain (VL + CL) or light chain variable domain (VL), or an antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) that is operably linked to the half-life extension element through an optionally protease-cleavable linker. When the antibody heavy chain constant region is on the first polypeptide, the antibody light chain is on the second polypeptide and when the antibody heavy chain constant region is on the second polypeptide, the antibody light chain is on the first polypeptide. The portion of the antibody heavy chain together with the complementary light chain associate to form a binding site for IL-10. In an example, the first polypeptide can comprise IL-10 and a portion of an antibody light chain that are linked through a protease-cleavable linker. The second polypeptide comprises a half-life extension element and a portion of an antibody heavy chain that is complementary to the antibody light chain that are linked through an optionally protease-cleavable linker. The portion of the antibody heavy chain together with the complementary light chain associate to form a binding site for IL-10. In another example, the first polypeptide can comprise IL-10 and a portion of an antibody heavy chain that are linked through a protease-cleavable linker. The second polypeptide can comprise a half-life extension element and a portion of an antibody light chain that are linked through an optionally protease-cleavable linker. The portion of the antibody heavy chain together with the complementary light chain associate to form a binding site for IL-10. 22 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0104] As disclosed herein, the inducible IL-10 prodrugs disclosed herein at least one blocking element, and optionally at least one half-life extension element, and such elements can contain two or more components that are present on the same polypeptide chain or on different polypeptide chains. The first polypeptide chain can comprise a first half-life extension element, and a second polypeptide chain can comprise a second half-life extension element. The first half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that optionally comprises one or more amino acid mutations that creates a “knob,” and the second half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that optionally comprises one or more amino acid mutations that create a “hole.” The first half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that optionally comprises one or more amino acid mutations that creates a “hole,” and the second half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that optionally comprises one or more amino acid mutations that create a “knob.” The first half-life extension element and the second half-life extension element can form a heterodimer (i.e., heterodimerize). The first half-life extension element and the second half-life extension element can form a heterodimer through disulfide bonds or a optionally protease cleavable linker, for example.

[0105] The first polypeptide chain and second polypeptide chain can each comprise a half-life extension element. For example, the first polypeptide chain can comprise the first half-life extension element, an IL- 10 polypeptide, and a blocking element, and the second polypeptide chain can comprise the second half- life extension element. For example, the first polypeptide chain can comprise the first half-life extension element and an IL-10 polypeptide, and the second polypeptide chain can comprise the second half-life extension element and a blocking element. For example, the first polypeptide chain can comprise the first half-life extension element and a blocking element, and the second polypeptide chain can comprise the second half-life extension element and a IL-10 polypeptide.

[0106] For example, the inducible IL-10 prodrug can have a first polypeptide of Formula XVII: [D]- [L2]-[H], and a second polypeptide of Formula XVIIII: [A]-[L1]-[H]. For example, the inducible IL-10 prodrug can have a first polypeptide of Formula XIX: [H]-[L2]-[D], and a second polypeptide of Formula XX: [H]-[L1]-[A]. In Formulas [XV]-[XVII], [A] is an IL-10 polypeptide, [D] is a blocking element, [H] is a half-life extension element, [L1] is a protease cleavable linker, and [L2] is an optionally protease cleavable linker.

[0107] In embodiments, each polypeptide chain can contain one IL-10 polypeptide, one half-life extension element, and one blocking element. For example, a first polypeptide chain can comprise an IL- 23 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 10 polypeptide, a half-life extension element, and a blocking element, and the second polypeptide chain can comprise an IL-10 polypeptide, a half-life extension element, and a blocking element.

[0108] The inducible IL-10 prodrugs disclosed herein can also comprise one or more additional blocking elements. For example, the first polypeptide chain can comprise one or more additional blocking elements. For example, the second polypeptide chain can comprise one or more additional blocking elements.

[0109] In some embodiments, the inducible IL-10 prodrug comprises: (i) IL-10; (ii) a half-life extension element selected from an antibody or fragment thereof (particularly a dAb), e.g., an antibody or fragment that binds serum albumin (particularly HSA), an immunoglobulin Fc or fragment thereof, or serum albumin (particularly HSA); (iii) a blocking element selected from an IL-10 binding Fab, dAb, scFv, or cognate IL-10 receptor or fragment thereof; and a protease cleavable linker. B. Exemplary Inducible IL-10 Prodrugs

[0110] The inducible IL-10 prodrugs comprise an IL-10 polypeptide, a half-life extension element, a blocking element, and a protease cleavable linker. Preferred inducible IL-10 prodrugs comprise two polypeptide chains.

[0111] The inducible IL-10 prodrugs can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 68, 72-76, 81-87, 93-104, 116-119, 123-126, and 514- 567, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NOs: 69, 70, 71, 568-621.

[0112] Compounds 1-93 and 97-100 are specific examples of inducible IL-10 prodrugs. Further details of exemplary IL-10 prodrugs are described in Table 10. Table 10. Inducible IL-10 prodrugs Inducible IL-10 First Polypeptide Second Polypeptide\\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Compound 8 SEQ ID NO: 521 SEQ ID NO: 575 Compound 9 SEQ ID NO: 522 SEQ ID NO: 576\\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Compound 40 SEQ ID NO: 553 SEQ ID NO: 607 Compound 41 SEQ ID NO: 554 SEQ ID NO: 608\\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Compound 72 SEQ ID NO: 82 SEQ ID NO: 69 Compound 73 SEQ ID NO: 83 SEQ ID NO: 69

[0113] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 68, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 68, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 27 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 68, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 72, and a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 73, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 74, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 74, and 2) a second polypeptide chain can comprises or consists the amino acid sequence of SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 74, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 75, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 75, and 2) a second polypeptide chain can that comprises or consists the amino acid sequence of SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 75, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 76, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 75, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 76, and 2) a second polypeptide chain can comprise or consist the amino acid sequence of SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 76, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 81, and 28 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 81, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 81, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 82, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 83, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 84, and 2) a second polypeptide chain can comprise or consist the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 85, and 20 a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 86, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 87, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 92, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 93, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 94, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 95, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 96, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can 29 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 97, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 98, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 99, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 100, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 101, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 102, and 2) a second polypeptide chain comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 103, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consist the amino acid sequence of SEQ ID NO: 104, and 2) polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 622, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69.

[0114] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consist the amino acid sequence of SEQ ID NO: 116, and 2) polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 117, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 118, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 119, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid 30 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 sequence of SEQ ID NO: 123, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 124, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 125, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 126, and 2) a second polypeptide chain that comprises or consists the amino acid sequence of SEQ ID NO: 69.

[0115] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 514, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 568.

[0116] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 515, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 569.

[0117] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 516, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 570.

[0118] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 517, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 571.

[0119] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 518, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 572.

[0120] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 519, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 573.

[0121] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 520, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 574.

[0122] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 521, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 575. 31 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0123] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 522, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 576.

[0124] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 523, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 577.

[0125] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 524, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 578.

[0126] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 525, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 579.

[0127] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 526, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 580.

[0128] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 527, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 581.

[0129] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 528, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 582.

[0130] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 529, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 583.

[0131] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 530, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 584.

[0132] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 531, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 585.

[0133] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 532, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 586. 32 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0134] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 533, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 587.

[0135] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 534, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 588.

[0136] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 535, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 589.

[0137] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 536, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 590.

[0138] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 537, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 591.

[0139] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 538, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 592.

[0140] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 539, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 593.

[0141] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 540, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 594.

[0142] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 541, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 595.

[0143] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 542, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 596.

[0144] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 543, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 597. 33 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0145] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 544, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 598.

[0146] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 545, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 599.

[0147] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 546, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 600.

[0148] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 547, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 601.

[0149] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 548, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 602.

[0150] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 549, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 603.

[0151] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 550, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 604.

[0152] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 551, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 605.

[0153] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 552, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 606.

[0154] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 553, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 607.

[0155] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 554, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 608. 34 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0156] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 555, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 609.

[0157] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 556, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 610.

[0158] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 557, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 611.

[0159] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 558, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 612.

[0160] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 559, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 613.

[0161] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 560, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 614.

[0162] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 561, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 615.

[0163] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 562, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 616.

[0164] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 563, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 617.

[0165] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 564, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 618.

[0166] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 565, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 619. 35 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0167] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 566, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 620.

[0168] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 567, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 621.

[0169] Amino acid sequence variants of Compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 ,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 97, 98, 99, or 100 that retain attenuated IL-10 activity in the periphery and that release active IL-10 upon protease cleavage in the tumor microenvironment can also be used in accordance with this disclosure.

[0170] For example, the inducible IL-10 prodrugs can include 1) a first polypeptide that comprises or consists of an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 68, 72-76, 81-87, 93-104, 116- 119, 123-126, and 514-567 and 2) a second polypeptide that comprises or consists of an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ IND NOs: 69, 70, 71, 568-621.

[0171] In some embodiments, the first polypeptide chain of the inducible IL-10 prodrug comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NOs.: 68, 69, 73-76, 81-87, 92-104, or 622, and the second polypeptide chain of the inducible IL-10 prodrug comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69, SEQ ID NO: 70, or SEQ ID NO: 71.

[0172] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 68, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least 36 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 68, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 68, and 2) a second polypeptide chain that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 72, and 2) a second polypeptide chain that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 73, and 2) a second polypeptide chain that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 74, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For 37 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 example, the inducible IL-10 prodrug can include a 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 74, and 2) a second polypeptide chain that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 74, and 2) a second polypeptide chain that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 75, and 2) a second polypeptide chain that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 75, and 2) a second polypeptide chain that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 75, and 2) the second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at 38 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 76, and 2) the second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 75, and 2) the second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 76, and 2) the second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 76, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 81, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at 39 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 least about 99% identical that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 81, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 70. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 81, and 2) the second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 71. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 82, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 83, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 84, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 40 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 85, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence of that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 86, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 87, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 92, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 93, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 94, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 41 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 95, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 96, an 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 97, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 98, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 99, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least 42 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 100, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 101, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 102, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 103, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 104, and 2) a second polypeptide chain comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid 43 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 622, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69.

[0173] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 116, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 117, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 118, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 119, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least 44 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 123, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 124, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 125, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69. For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 126, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 69.

[0174] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 531, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 585.

[0175] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, 45 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 532, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 586.

[0176] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 533, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 587.

[0177] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 534, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 588.

[0178] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 535, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 589.

[0179] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 536, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 590. 46 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0180] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 540, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 594.

[0181] For example, the inducible IL-10 prodrug can include 1) a first polypeptide that comprises an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 542, and 2) a second polypeptide chain comprises the amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 596. C. Blocking Element

[0182] The blocking element can be any element that inhibits the ability of IL-10 to bind and / or activate its receptor e.g., by sterically inhibiting the binding of IL-10 within the prodrug to the IL-10 receptor and / or by noncovalently binding to IL-10 within the prodrug and inhibiting its binding to the IL-10 receptor. The blocking element disclosed herein can bind to IL-10. Examples of suitable blocking elements include a chain of the cognate receptor of IL-10 and an IL-10-binding fragment or mutein thereof. The blocking element can include one or both of the IL10Ralpha or a portion thereof and / or one or both of the IL10Rbeta or a portion thereof.

[0183] Antibodies and antigen-binding fragments thereof that bind IL-10, including an antigen-binding fragment (Fab), a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody a single chain variable fragment (scFv), single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL) and a variable domain of camelid-type nanobody (VHH), a dAb and the like, can also be used as blocking elements. Other suitable antigen-binding domains that bind IL- 10 can also be used, including non-immunoglobulin proteins that mimic antibody binding and / or structure such as, anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, DARPins, fynomers, kunitz domain peptides, monobodies, and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin and CTLA4 scaffolds. 47 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0184] Further examples of suitable blocking polypeptides include polypeptides that sterically inhibit or block binding of IL-10 to its cognate receptor. Advantageously, such moieties can also function as half- life extending elements. For example, a peptide that is modified by conjugation to a water-soluble polymer, such as PEG, can sterically inhibit or prevent binding of the cytokine to its receptor. Polypeptides, or fragments thereof, that have long serum half-lives can also be used, such as serum albumin (human serum albumin), immunoglobulin Fc, transferrin and the like, as well as fragments and muteins of such polypeptides. Similarly, proteins or polypeptides, such as antibodies of antibody fragments, which bind such proteins, such as an scFv or single variable domain that binds human serum albumin can be used as blocking elements.

[0185] IL-10 blocking elements that are particularly suitable are single chain variable fragments (scFv), Fab fragments, or full length or an IL-10-binding fragment or mutein of the cognate receptor of an IL-10 (e.g., IL10Ralpha or IL10Rbeta). Typically, the IL-10 blocking element will be an IL-10-binding Fab, dAb, scFv, or cognate IL-10 receptor or fragment thereof.

[0186] Also disclosed herein is an inducible IL-10 polypeptide that contains a blocking element having specificity for IL-10 and further contains a separate half-life extension element.

[0187] The blocking element can contain two or more components that are present on the same polypeptide chain or on separate polypeptide chains. For example, A first polypeptide chain can include an antibody light chain (VL+CL), or light chain variable domain (VL) and a second polypeptide can include an antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) that is complementary to the VL+ CL or VL on the first polypeptide. In such situations, these components can associate in the IL-10 prodrug (as a complex) to form an antigen-binding site, such as an Fab that binds IL-10 and attenuates IL-10 activity.

[0188] Blocking elements 1-41 are specific examples of blocking elements comprising a heavy chain variable domain and a light chain variable domain. Further details of exemplary IL-10 antibody blocking elements are described in Table 8. Table 8. Blocking Elements Blocking Element Heavy chain variable Light chain variable domain\\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Blocking Element 5 SEQ ID NO: 131 SEQ ID NO: 172 Blocking Element 6 SEQ ID NO: 132 SEQ ID NO: 173\\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Blocking Element 37 SEQ ID NO: 163 SEQ ID NO: 509 Blocking Element 38 SEQ ID NO: 164 SEQ ID NO: 510 [of the amino acid sequence of SEQ ID NO: 127, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 127, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 168, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 168.

[0190] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 128, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 128, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 169, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 169.

[0191] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 129, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 129, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 170, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 170.

[0192] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 130, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 130, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 171, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 171.

[0193] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 131, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 131, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 172, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 172.

[0194] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 132, or an amino acid sequence that has at least 95% identity to 50 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 SEQ ID NO: 132, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 173, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 173.

[0195] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 133, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 133, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 174, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 174.

[0196] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 134, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 134, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 175, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 175.

[0197] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 135, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 135, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 176, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 176.

[0198] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 136, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 136, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 177, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 177.

[0199] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 137, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 137, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 178, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 178.

[0200] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 138, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 138, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 179, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1789. 51 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0201] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 139, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 139, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 180, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 180.

[0202] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 140, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 140, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 181, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 181.

[0203] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 141, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 141, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 182, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 182.

[0204] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 142, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 142, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 183, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 183.

[0205] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 143, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 143, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 184, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 184.

[0206] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 144, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 143, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 185, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 185.

[0207] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 145, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 145, and 2) a second polypeptide chain that comprises or consists of the amino acid 52 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 sequence of SEQ ID NO: 186, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 186.

[0208] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 146, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 146, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 187, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 187.

[0209] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 147, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 147, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 188, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 188.

[0210] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 148, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 148, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 189, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 189.

[0211] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 149, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 149, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 190, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 190.

[0212] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 150, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 150, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 191, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 191.

[0213] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 151, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 151, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 192, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 192. 53 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0214] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 152, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 152, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 193, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 193.

[0215] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 153, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 153, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 194, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 194.

[0216] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 154, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 154, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 500, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 500.

[0217] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 155, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 155, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 501, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 501.

[0218] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 156, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 156, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 502, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 502.

[0219] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 157, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 157, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 503, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 503.

[0220] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 158, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 158, and 2) a second polypeptide chain that comprises or consists of the amino acid 54 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 sequence of SEQ ID NO: 504, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 504.

[0221] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 159, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 159, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 505, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 505.

[0222] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 160, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 160, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 507, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 507.

[0223] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 161, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 161, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 508, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 508.

[0224] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 162, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 162, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 509, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 509.

[0225] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 163, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 163, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 510, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 510.

[0226] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 164, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 164, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 511, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 511. 55 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0227] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 165, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 165, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 512, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 511.

[0228] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 166, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 166, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 512, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 512.

[0229] The blocking element can comprise 1) a first polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 167, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 167, and 2) a second polypeptide chain that comprises or consists of the amino acid sequence of SEQ ID NO: 513, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 513. D. Half-Life Extension Element

[0230] The half-life extension element increases the in vivo half-life and provides altered pharmacodynamics and pharmacokinetics of the inducible IL-10 prodrugs. Without being bound by theory, the half-life extension element alters pharmacodynamic properties including alteration of tissue distribution, penetration, and diffusion of the inducible IL-10 prodrug. In some embodiments, the half-life extension element can improve tissue targeting, tissue penetration, diffusion within the tissue, and enhanced efficacy as compared with a protein without a half-life extension element. Without being bound by theory, an exemplary way to improve the pharmacokinetics of a polypeptide is by expression of an element in the polypeptide chain that binds to receptors that are recycled to the plasma membrane of cells rather than degraded in the lysosomes, such as the FcRn receptor on endothelial cells and transferrin receptor. Three types of proteins, e.g., human IgGs, HSA (or fragments), and transferrin, persist for much longer in human serum than would be predicted just by their size, which is a function of their ability to bind to receptors that are recycled rather than degraded in the lysosome. These proteins, or fragments that retain FcRn binding, are routinely linked to other polypeptides to extend their serum half-life. HSA may also be directly bound to the pharmaceutical compositions or bound via a short linker. Fragments of HSA may also be used. HSA and fragments thereof can function as both a blocking element and a half-life extension element. Human IgGs and Fc fragments can also carry out a similar function. 56 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0231] The serum half-life extension element can also be an antigen-binding polypeptide that binds to a protein with a long serum half-life such as serum albumin (e.g., HSA), transferrin and the like. Examples of such polypeptides include antibodies and fragments thereof including, a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a single chain variable fragment (scFv), an antigen binding fragment (Fab), single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL) and a variable domain of camelid-type nanobody (VHH), a dAb and the like. Other suitable antigen-binding domains include non-immunoglobulin proteins that mimic antibody binding and / or structure such as, anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, DARPins, fynomers, kunitz domain peptides, monobodies, and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin and CTLA4 scaffolds. Further examples of antigen-binding polypeptides include a ligand for a desired receptor, a ligand-binding portion of a receptor, a lectin, and peptides that bind to or associate with one or more target antigens. The antibodies and fragments thereof can function as both a blocking element and a half-life extension element.

[0232] The half-life extension element can also function as both a blocking element and a half-life extension element. For instance, the half-life extension element (e.g., HSA binding polypeptide) can function as a blocking element (that sterically inhibits binding of IL-10 to the IL-10 receptor), for example, when adjacent to the IL-10 polypeptide.

[0233] The half-life extension element as provided herein is preferably a human serum albumin (HSA), an antigen binding polypeptide that binds human serum albumin, or an immunoglobulin Fc or fragment thereof. The half-life extension element can comprise an Fc domain or a fragment thereof. For example, the Fc domain can comprise a CH2 and CH3 domain or fragment thereof. For example, the Fc domain can comprise a constant domain of the heavy chain polypeptide. Typically, the half-life extension element is an antibody or fragment thereof (particularly a dAb), e.g., an antibody or fragment that binds serum albumin (particularly HSA), an immunoglobulin Fc or fragment thereof, or serum albumin (particularly HSA).

[0234] The half-life extension element disclosed herein can comprise a first half-life extension element and a second half-life extension element. The first and second half-life extension element can heterodimerize. The first and second half-life extension elements can include one or more modifications that promote heterodimerization of the first and second half-life extension elements. For example, one or more amino acid modifications can be made to the first half-life extension element. For example, one or more amino acid modifications can be made to the second half-life extension element. The inducible IL- 10 prodrugs disclosed herein can comprise a first half-life extension element and a second half-life 57 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 extension element, each of which comprises a CH3 domain. In embodiments, the half-life extension element comprising a CH3 domain is a heavy chain polypeptide or a fragment thereof (e.g., an Fc domain or fragment thereof). The CH3 domains of the first and second half-life extension elements can be altered by the “knobs-into-holes” technology. See, e.g., WO1996 / 027011; Ridgway et al., Protein Eng., 9(7):617- 621 (1996); Merchant et al., Nat. Biotechnol., 16(7):677-681 (1998). Using the knob-into-holes method, the interaction surfaces of the two CH3 domains are altered to increase the heterodimerization of the first half-life extension element and the second half-life extension element each containing an altered CH3 domain. This occurs by introducing a bulky residue into the CH3 domain of one of the half-life extension elements, which acts as the “knob.” Then, in order to accommodate the bulky residue, a “hole” is formed in the other half-life extension domain that can accommodate the knob. Either of the altered CH3 domains can be the “knob” while the other can be the “hole.” The introduction of a disulfide bridge can further stabilize the heterodimers.

[0235] In embodiments, the knobs-into-holes approach can be used to promote heterodimerization between two different half-life extension elements (e.g., between a first half-life extension element and a second half-life extension element).

[0236] In embodiments, the IL-10 prodrugs described herein can comprise two half-life extension elements (i.e., a first half-life element and a second-half-life extension element). In embodiments, the first half-life extension element and the second half-life extension element can be linked via a linker. The first half-life extension element and the second half-life extension element can be an Fc domain or fragment thereof. The first half-life extension element and the second half-life extension element can be an antibody, or a fragment, variant, or derivative thereof.

[0237] The first half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., the heavy chain constant regions of an Fc domain, or fragment thereof) that comprises one or more amino acid mutations that creates a “knob,” and the second half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., the heavy chain constant regions of an Fc domain, or fragment thereof) that comprises one or more amino acid mutations that create a “hole.” The first half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that comprises one or more amino acid mutations that creates a “hole,” and the second half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that comprises one or more amino acid mutations that create a “knob.” The techniques and procedures for introducing a hole or knob in a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) are known by those skilled in the art. 58 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0238] In embodiments, the first half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that comprises the amino acid mutations S354C and T366W (numbered according the Kabat EU numbering system), and the second half-life extension element comprises a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that comprises the amino acid mutations Y349C, T366S, L368A, and Y407V (numbered according to the Kabat EU numbering system). In embodiments, the first half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that comprises the amino acid mutations Y349C, T366S, L368A, and Y407V (numbered according to the Kabat EU numbering system), and the second half-life extension element can comprise a heavy chain polypeptide or portion thereof (e.g., an Fc domain or fragment thereof) that comprises the amino acid mutations S354C and T366W (numbered according to the Kabat EU numbering system).

[0239] In embodiments, any of the following amino acid substitutions can be made to a first half-life extension element and a paired second half-life extension element that each contain an Fc domain: (a) Y407T in the first half-life extension and T366Y in the second half-life extension element; (b) Y407A in the first half-life extension element and T366W in the second half-life extension element; (c) F405A in the first half-life extension element and T394W in the second half-life extension element; (d) F405W in the first half-life extension element and T394S in the second half-life extension element; (e) Y407T in the first half-life extension element and T366Y in the second half-life extension element; (f) T366Y and F405A in the first half-life extension element and T394W and Y407T in the second half-life extension element; (g) T366W and F405W in the first half-life extension element and T394S and Y407A in the second half-life extension element; (h) F405W and Y407A in the first half-life extension element and T366W and T394S in the second half-life extension element; or (i) T366W in the first half-life extension element and T366S, L368A, and Y407V in the second half-life extension element, numbered according to the Kabat EU numbering system.

[0240] In embodiments, any of the following amino acid substitutions can be made to a first half-life extension element and a paired second half-life extension element that each contain an Fc domain: (a) Y407T in the second half-life extension element and T366Y in the first half-life extension element; (b) Y407A in the second half-life extension element and T366W in the first half-life extension element; (c) F405A in the second half-life extension element and T394W in the first half-life extension element; (d) F405W in the second half-life extension element and T394S in the first half-life extension element; (e) Y407T in the second half-life extension element and T366Y in the first half-life extension element; (f) T366Y and F405A in the second half-life extension element and T394W and Y407T in the first half-life extension element; (g) T366W and F405W in the second half-life extension element and T394S and 59 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Y407A in the first half-life extension element; (h) F405W and Y407A in the second half-life extension element and T366W and T394S in the first half-life extension element; or (i) T366W in the second half- life extension element and T366S, L368A, and Y407V in the first half-life extension element, numbered according to the Kabat EU numbering system.

[0241] In some embodiments, the half-life extension element disclosed herein can comprise a heavy chain polypeptide that comprises one or more amino acid substitutions altering effector function. In some embodiments, the half-life extension element is an IgG1 heavy chain polypeptide and comprises the amino substitution(s): N297A, N297G, or N297Q; L234A and L235A; C220S, C226S, C229S, and P238S; C226S, C229S, E233P, L234V, and L235A; L234F, L235E, and P331S; S267E and L328F; D265A; and / or L234A, L235A, and P329G, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element disclosed herein is an IgG1 heavy chain polypeptide and comprises one or more amino acid substitutions selected from the group consisting of N297A, N297G, N297Q, L234A, L235A, C220S, C226S, C229S, P238S, E233P, L234V, L234F, L235E, P331S, S267E, L328F, D265A, and P329G, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element disclosed herein is an IgG2 heavy chain polypeptide and comprises the amino substitution(s): V234A and G237A; H268Q, V309L, A330S, and A331S; or V234A, G237A, P238S, H268A, V309L, and A330S, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element disclosed herein is an IgG2 heavy chain polypeptide and comprises one or more amino acid substitutions selected from the group consisting of V234A, G237A, H268Q, V309L, A330S, A331S, P238S, H268A, and V309L, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element disclosed herein is an IgG4 heavy chain polypeptide and comprises the amino substitution(s): L235A, G237A, and E318A; S228P, L234A, and L235A; H268Q, V309L, A330S, and P331S; and / or S228P and L235A, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element disclosed herein is an IgG2 heavy chain polypeptide and comprises one or more amino acid substitutions selected from the group consisting of L235A, G237A, E318A, S228P, L234A, H268Q, V309L, A330S, and P331S, numbered according to the Kabat EU numbering system.

[0242] In some embodiments, the half-life extension element disclosed herein comprises a heavy chain polypeptide that comprises one or more amino acid substitutions enhancing effector function. In some embodiments, the half-life extension element is an IgG1 heavy chain polypeptide and comprises the amino acid substitution(s): S298A, E333A, and K334A; S239D and I332E; S239D, A330L, and I332E; P247I and A339D or A339Q; D280H and K290S; D280H, K290S, and either S298D or S298V; F243L, R292P, and Y300L; F243L, R292P, Y300L, and P396L; F243L, R292P, Y300L, V305I, and P396L; 60 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 G236A, S239D, and I332E; K326A and E333A; K326W and E333S; K290E, S298G, and T299A; K290E, S298G, T299A, and K326E; K290N, S298G, and T299A; K290N, S298G, T299A, and K326E; K334V; L235S, S239D, and K334V; K334V and Q331M, S239D, F243V, E294L, or S298T; E233L, Q311M, and K334V; L234I, Q311M, and K334V; K334V and S298T, A330M, or A330F; K334V, Q311M, and either A330M or A330F; K334V, S298T, and either A330M or A330F; K334V, S239D, and either A330M or S298T; L234Y, Y296W, and K290Y, F243V, or E294L; Y296W and either L234Y or K290Y; S239D, A330S, and I332E, V264I; F243L and V264I; L328M; I332E; L328M and I332E; V264I and I332E; S239E and I332E; S239Q and I332E; S239E; A330Y; I332D; L328I and I332E; L328Q and I332E; V264T; V240I; V266I; S239D; S239D and I332D; S239D and I332N; S239D and I332Q; S239E and I332D; S239E and I332N; S239E and I332Q; S239N and I332D; S239N and I332E; S239Q and I332D; A330Y and I332E; V264I, A330Y, and I332E; A330L and I332E; V264I, A330L, and I332E; L234E, L234Y, or L234I; L235D, L235S, L235Y, or L235I; S239T; V240M; V264Y; A330I; N325T; I332E and L328D, L328V, L328T, or L328I; V264I, I332E, and either S239E or S239Q; S239E, V264I, A330Y, and I332E; A330Y, I332E, and either S239D or S239N; A330L, I332E, and either S239D or S239N; V264I, S298A, and I332E; S298A, I332E, and either S239D or S239N; S239D, V264I, and I332E; S239D, V264I, S298A, and I332E; S239D, V264I, A330L, and I332E; S239D, I332E, and A330I; P230A; P230A, E233D, and I332E; E272Y; K274T, K274E, K274R, K274L, or K274Y; F275W; N276L; Y278T; V302I; E318R; S324D, S324I or S324V; K326I or K326T; T335D, T335R, or T335Y; V240I and V266I; S239D, A330Y, I332E, and L234I; S239D, A330Y, I332E, and L235D; S239D, A330Y, I332E, and V240I; S239D, A330Y, I332E, and V264T; and / or S239D, A330Y, I332E, and either K326E or K326T, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element is an IgG1 heavy chain polypeptide and comprises one or more amino acid substitution(s) selected from the group consisting of: P230A, E233D, L234E, L234Y, L234I, L235D, L235S, L235Y, L235I, S239D, S239E, S239N, S239Q, S239T, V240I, V240M, F243L, V264I, V264T, V264Y, V266I, E272Y, K274T, K274E, K274R, K274L, K274Y, F275W, N276L, Y278T, V302I, E318R, S324D, S324I, S324V, N325T, K326I, K326T, L328M, L328I, L328Q, L328D, L328V, L328T, A330Y, A330L, A330I, I332D, I332E, I332N, I332Q, T335D, T335R, and T335Y.

[0243] In some embodiments, the half-life extension element comprises an Fc domain or fragment thereof that comprises one or more amino acid substitutions altering effector function. In some embodiments, the half-life extension element is an IgG1 Fc element or fragment thereof and comprises the amino substitution(s): N297A, N297G, or N297Q; F234A and F235A; C220S, C226S, C229S, and P238S; C226S, C229S, E233P, F234V, and F235A; F234F, F235E, and P331S; S267E and F328F; D265A; and / or F234A, F235A, and P329G, numbered according to the Rabat EU numbering system. In 61 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 some embodiments, the half-life extension element is an IgG1 Fc element or fragment thereof and comprises one or more amino acid substitutions selected from the group consisting of N297A, N297G, N297Q, L234A, L235A, C220S, C226S, C229S, P238S, E233P, L234V, L234F, L235E, P331S, S267E, L328F, D265A, and P329G, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element is an IgG2 Fc element or fragment thereof and comprises the amino substitution(s): V234A and G237A; F1268Q, V309L, A330S, and A331S; and / or V234A, G237A, P238S, F1268A, V309L, and A330S, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element is an IgG2 Fc element or fragment thereof and comprises one or more amino acid substitutions selected from the group consisting of V234A, G237A, F1268Q, V309L, A330S, A331S, P238S, F1268A, and V309L, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element is an IgG4 Fc element or fragment thereof and comprises the amino substitution(s): L235A, G237A, and E318A; S228P, L234A, and L235A; F1268Q, V309L, A330S, and P331S; and / or S228P and L235A, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element is an IgG2 Fc element or fragment thereof and comprises one or more amino acid substitutions selected from the group consisting of L235A, G237A, E318A, S228P, L234A, F1268Q, V309L, A330S, and P331S, numbered according to the Kabat EU numbering system.

[0244] In some embodiments, the half-life extension element comprises Fc domain? or fragment thereof that comprises one or more amino acid substitutions enhancing effector function. In some embodiments, the half-life extension element is an IgG1 Fc element or fragment thereof and comprises the amino acid substitution(s): S298A, E333A, and K334A; S239D and I332E; S239D, A330L, and I332E; P247I and A339D or A339Q; D280H and K290S; D280H, K290S, and either S298D or S298V; F243L, R292P, and Y300L; F243L, R292P, Y300L, and P396L; F243L, R292P, Y300L, V305I, and P396L; G236A, S239D, and I332E; K326A and E333A; K326W and E333S; K290E, S298G, and T299A; K290E, S298G, T299A, and K326E; K290N, S298G, and T299A; K290N, S298G, T299A, and K326E; K334V; L235S, S239D, and K334V; K334V and Q331M, S239D, F243V, E294L, or S298T; E233L, Q311M, and K334V; L234I, Q311M, and K334V; K334V and S298T, A330M, or A330F; K334V, Q311M, and either A330M or A330F; K334V, S298T, and either A330M or A330F; K334V, S239D, and either A330M or S298T; L234Y, Y296W, and K290Y, F243V, or E294L; Y296W and either L234Y or K290Y; S239D, A330S, and I332E, V264I; F243L and V264I; L328M; I332E; L328M and I332E; V264I and I332E; S239E and I332E; S239Q and I332E; S239E; A330Y; I332D; L328I and I332E; L328Q and I332E; V264T; V240I; V266I; S239D; S239D and I332D; S239D and I332N; S239D and I332Q; S239E and I332D; S239E and I332N; S239E and I332Q; S239N and I332D; S239N and I332E; S239Q and I332D; 62 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 A330Y and I332E; V264I, A330Y, and I332E; A330L and I332E; V264I, A330L, and I332E; L234E, L234Y, or L234I; L235D, L235S, L235Y, or L235I; S239T; V240M; V264Y; A330I; N325T; I332E and L328D, L328V, L328T, or L328I; V264I, I332E, and either S239E or S239Q; S239E, V264I, A330Y, and I332E; A330Y, I332E, and either S239D or S239N; A330L, I332E, and either S239D or S239N; V264I, S298A, and I332E; S298A, I332E, and either S239D or S239N; S239D, V264I, and I332E; S239D, V264I, S298A, and I332E; S239D, V264I, A330L, and I332E; S239D, I332E, and A330I; P230A; P230A, E233D, and I332E; E272Y; K274T, K274E, K274R, K274L, or K274Y; F275W; N276L; Y278T; V302I; E318R; S324D, S324I or S324V; K326I or K326T; T335D, T335R, or T335Y; V240I and V266I; S239D, A330Y, I332E, and L234I; S239D, A330Y, I332E, and L235D; S239D, A330Y, I332E, and V240I; S239D, A330Y, I332E, and V264T; and / or S239D, A330Y, I332E, and either K326E or K326T, numbered according to the Kabat EU numbering system. In some embodiments, the half-life extension element is an IgGl Fc domain or fragment thereof and comprises one or more amino acid substitution(s) selected from the group consisting of: P230A, E233D, L234E, L234Y, L234I, L235D, L235S, L235Y, L235I, S239D, S239E, S239N, S239Q, S239T, V240I, V240M, F243L, V264I, V264T, V264Y, V266I, E272Y, K274T, K274E, K274R, K274L, K274Y, F275W, N276L, Y278T, V302I, E318R, S324D, S324I, S324V, N325T, K326I, K326T, L328M, L328I, L328Q, L328D, L328V, L328T, A330Y, A330L, A330I, I332D, I332E, I332N, I332Q, T335D, T335R, and T335Y.

[0245] In some embodiments, the half-life extension element comprises one or more amino acid substitution(s) that enhance binding of the half-life extension element to FcRn. In some embodiments, the one or more amino acid substitution(s) increase binding affinity of an Fc-containing polypeptide (e.g., a heavy chain polypeptide or an Fc element or fragment thereof) to FcRn at acidic pH. In some embodiments, the half-life extension element comprises one or more amino acid substitution(s) selected from the group consisting of M428F; T250Q and M428F; M252Y, S254T, and T256E; P257I and N434H; D376V and N434H; P257I and Q3111; N434A; N434W; M428F and N434S; V259I and V308F; M252Y, S254T, and T256E; V259I, V308F and M428F; T307Q and N434A; T307Q and N434S; T307Q, E380A, and N434A; V308P and N434A; N434H; and V308P, numbered according to the Kabat EU numbering system?.

[0246] The half-life extension element of an inducible IL-10 prodrug extends the half-life of the inducible IL-10 prodrug by at least about two days, about three days, about four days, about five days, about six days, about seven days, about eight days, about nine days, about 10 days or more. E. Protease Cleavable Linker 63 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0247] As disclosed herein, the inducible IL-10 prodrug comprises one or more linker sequences. A linker sequence serves to provide flexibility between the polypeptides, such that, for example, the blocking element is capable of inhibiting the activity of IL-10. The linker can be located between the IL- 10 polypeptide, the half-life extension element, and / or the blocking element. As described herein the inducible IL-10 prodrug comprises a protease cleavable linker. The protease cleavable linker can comprise one or more cleavage sites for one or more desired proteases. Preferably, the desired protease is enriched or selectively expressed at the desired target site of IL-10 activity (e.g., a site of inflammation). Thus, the inducible IL-10 prodrug is preferentially or selectively cleaved at the target site of desired IL-10 activity.

[0248] Suitable linkers are typically less than about 100 amino acids. Such linkers can be of different lengths, such as from 1 amino acid (e.g., Gly) to 30 amino acids, from 1 amino acid to 40 amino acids, from 1 amino acid to 50 amino acids, from 1 amino acid to 60 amino acids, from 1 to 70 amino acids, from 1 to 80 amino acids, from 1 to 90 amino acids, and from 1 to 100 amino acids. In some embodiments, the linker is at least about 1, about 2, about 3, about 4, about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, or about 100 amino acids in length. Preferred linkers are typically from about 5 amino acids to about 35 amino acids. Preferably the lengths of linkers vary from 2 to 35 amino acids, optimized for each condition so that the linker does not impose any constraints on the conformation or interactions of the linked domain. In a preferred embodiment, the linker is cleavable by a cleaving agent, e.g., an enzyme. Preferably, the linker comprises a protease cleavage site. In some cases, the linker comprises one or more cleavage sites. The linker can comprise a single protease cleavage site. The linker can also comprise 2 or more protease cleavage sites. For example, 2 cleavage sites, 3 cleavage sites, 4, cleavage sites, 5 cleavage sites, or more. In cases the linker comprises 2 or more protease cleavage sites, the cleavage sites can be cleaved by the same protease or different proteases. A linker comprising two or more cleavage sites is referred to as a “tandem linker.” The two or more cleavage sites can be arranged in any desired orientation, including, but not limited to one cleavage site adjacent to another cleavage site, one cleavage site overlapping another cleavage site, or one cleavage site following by another cleavage site with intervening amino acids between the two cleavage sites.

[0249] Of particular interest in the present invention are disease specific protease-cleavable linkers. Also preferred are protease-cleavable linkers that are preferentially cleaved at a desired location in the body, such as the inflammatory, relative to the peripheral circulation. For example, the rate at which the protease-cleavable linker is cleaved in the tumor microenvironment can be at least about 10 times, at least about 100 times, at least about 1000 times or at least about 10,000 times faster in the desired location in 64 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 the body, e.g., the tumor microenvironment, in comparison to in the peripheral circulation (e.g., in plasma).

[0250] Proteases known to be associated with diseased cells or tissues include but are not limited to serine proteases, cysteine proteases, aspartate proteases, threonine proteases, glutamic acid proteases, metalloproteases, asparagine peptide lyases, serum proteases, cathepsins, Cathepsin B, Cathepsin C, Cathepsin D, Cathepsin E, Cathepsin G, Cathepsin S, Cathepsin K, Cathepsin L, kallikreins, hKl, hK10, hK15, plasmin, collagenase, Type IV collagenase, stromelysin, Factor Xa, chymotrypsin-like protease, chymase, granzyme M, mast-cell carboxypeptidase (MC-CPA), trypsin-like protease, elastase (e.g., a neutrophil elastase), elastase-like protease, subtilisin-like protease, actinidain, bromelain, calpain, caspases, caspase-3, Mirl-CP, papain, HIV-1 protease, HSV protease, CMV protease, chymosin, renin, pepsin, matriptase, legumain, plasmepsin, nepenthesin, metalloexopeptidases, metalloendopeptidases, matrix metalloproteases (MMP), MMP1, MMP2, MMP3, MMP8, MMP9, MMP13, MMP11, MMP14, MMP19, MMP20, urokinase plasminogen activator (uPA), enterokinase, proteinase (e.g. proteinase 3), prostate-specific antigen (PSA, hK3), interleukin-1β converting enzyme, thrombin, FAP (FAP ^), dipeptidyl peptidase, meprins, granzymes and dipeptidyl peptidase IV (DPPIV / CD26). Proteases capable of cleaving linker amino acid sequences (which can be encoded by the chimeric nucleic acid sequences provided herein) can, for example, be selected from the group consisting of a prostate specific antigen (PSA), a matrix metalloproteinase (MMP), an A Disintigrin and a Metalloproteinase (ADAM), a plasminogen activator, a cathepsin, a caspase, and a tumor cell surface protease. The MMP can, for example, be matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), matrix metalloproteinase 14 (MMP14), matrix metalloproteinase 19 (MMP19), or matrix metalloproteinase 20 (MMP20). In addition, or alternatively, the linker can be cleaved by a cathepsin, such as, Cathepsin B, Cathepsin C, Cathepsin D, Cathepsin S, Cathepsin E, Cathepsin G, Cathepsin K and / or Cathepsin L. Preferably, the linker can be cleaved by MMP14, Cathepsin L, ADAM, FAP, proteinase 3, neutrophil elastase or Cathepsin G.

[0251] Proteases useful for cleavage of linkers and for use in cleaving the inducible IL-10 polypeptide prodrug disclosed herein are presented in Table 1, and exemplary proteases and their cleavage site are presented in Table 2. Table 1. Proteases relevant to inflammation and cancer Protease Specificity Other aspects65 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Protease Specificity Other aspects Granzyme B (grB) Cleaves after Asp Type of serine protease; strongly r id ( - ) im li td in ind in rf rin-d ndnt l h e; ; e e n\\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Protease Specificity Other aspects PAI-1, RANTES, TGFβ, leukocytes; regulates inflammatory TNF r n r m t t i n TProtease Cleavage Domain Sequence SEQ ID NO: MMP7 KRALGLPG\\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Protease Cleavage Domain Sequence SEQ ID NO: Thrombin PPRSFL 405 C 3 DEVD 406 [0, ers, thrombin cleavable linkers, chymase cleavable linkers, carboxypeptidase A cleavable linkers, cathepsin cleavable linkers, elastase cleavable linkers, FAP cleavable linkers, ADAM cleavable linkers, PR-3 cleavable linkers, granzyme M cleavable linkers, a calpain cleavable linkers, a matrix metalloproteinase (MMP) cleavable linkers, a plasminogen activator cleavable linkers, a caspase cleavable linkers, a tryptase cleavable linkers, or a tumor cell surface protease. Specifically, MMP9 cleavable linkers, ADAM cleavable linkers, CTSL1 cleavable linkers, FAPα cleavable linkers, and cathepsin cleavable linkers. Some preferred protease-cleavable linkers are cleaved by a MMP, a cathepsin, a FAPalpha, a ADAM, a neutrophil elastase, a proteinase 3.

[0253] The linker sequences disclosed herein are typically less than 100 amino acids. Such linker sequences can be of different lengths, such as from 1 amino acid (e.g., Gly) to 30 amino acids, from 1 amino acid to 40 amino acids, from 1 amino acid to 50 amino acids, from 1 amino acid to 60 amino acids, from 1 to 70 amino acids, from 1 to 80 amino acids, from 1 to 90 amino acids, and from 1 to 100 amino acids. In some embodiments, the linker is at least about 1, about 2, about 3, about 4, about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, or about 100 amino acids in length. Preferred linkers are typically from about 5 amino acids to about 35 amino acids.

[0254] Preferably the lengths of linkers vary from 2 to 35 amino acids, optimized for each condition so that the linker does not impose any constraints on the conformation or interactions of the linked domains. 68 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0255] In some embodiments, the linker comprises the sequence GPAGLYAQ (SEQ ID NO: 195); GPAGMKGL (SEQ ID NO: 196); PGGPAGIG (SEQ ID NO: 197); ALFKSSFP (SEQ ID NO: 198); ALFFSSPP (SEQ ID NO: 199); LAQRLRSS (SEQ ID NO: 200); LAQKLKSS (SEQ ID NO; 201); GALFKSSFPSGGGPAGLYAQGGSGKGGSGK (SEQ ID NO: 202); RGSGGGPAGLYAQGSGGGPAGLYAQGGSGK (SEQ ID NO: 203); KGGGPAGLYAQGPAGLYAQGPAGLYAQGSR (SEQ ID NO: 204); RGGPAGLYAQGGPAGLYAQGGGPAGLYAQK (SEQ ID NO: 205); KGGALFKSSFPGGPAGIGPLAQKLKSSGGS (SEQ ID NO: 206); SGGPGGPAGIGALFKSSFPLAQKLKSSGGG (SEQ ID NO: 207); RGPLAQKLKSSALFKSSFPGGPAGIGGGGK (SEQ ID NO: 208); GGGALFKSSFPLAQKLKSSPGGPAGIGGGR (SEQ ID NO: 209); RGPGGPAGIGPLAQKLKSSALFKSSFPGGG (SEQ ID NO: 210); RGGPLAQKLKSSPGGPAGIGALFKSSFPGK (SEQ ID NO: 211); RSGGPAGLYAQALFKSSFPLAQKLKSSGGG (SEQ ID NO: 212); GGPLAQKLKSSALFKSSFPGPAGLYAQGGR (SEQ ID NO: 213); GGALFKSSFPGPAGLYAQPLAQKLKSSGGK (SEQ ID NO: 214); RGGALFKSSFPLAQKLKSSGPAGLYAQGGK (SEQ ID NO: 215); RGGGPAGLYAQPLAQKLKSSALFKSSFPGG (SEQ ID NO: 216); SGPLAQKLKSSGPAGLYAQALFKSSFPGSK (SEQ ID NO: 217); KGGPGGPAGIGPLAQRLRSSALFKSSFPGR (SEQ ID NO: 218); KSGPGGPAGIGALFFSSPPLAQKLKSSGGR (SEQ ID NO: 219); SGGFPRSGGSFNPRTFGSKRKRRGSRGGGG (SEQ ID NO: 220); PGGPAGIGALFKSSFPPLAQKLKSS (SEQ ID NO: 5); LAQAVKSS (SEQ ID NO: 9); TEGEARGN (SEQ ID NO: 10); EYAEFKGT (SEQ ID NO: 11); GPLGLRAQ (SEQ ID NO: 14); TSGPNQEQ (SEQ ID NO: 15); TSGPAQEQ (SEQ ID NO: 16); PGGPAGIGALFKSSFPPLAQRLRSS (SEQ ID NO: 17); PGGPAGIGALFFSSPPPLAQKLKSS (SEQ ID NO: 18); GPAGLYAQALFKSSFPPLAQKLKSS (SEQ ID NO: 19); GPAGLYAQALFFSSPPPLAQKLKSS (SEQ ID NO: 20); GPAGLYAQPLAQKLKSS (SEQ ID NO: 21); GPAGLYAQGPAGLYAQGPAGLYAQ (SEQ ID NO: 22); GPAGLYAQPLAQKLKSSALFFSSPP (SEQ ID NO: 23); PLAQKLKSSALFFSSPPGPAGLYAQ (SEQ ID NO: 24); PLAQKLKSSGPAGLYAQALFFSSPP (SEQ ID NO: 25); PLAQRLRSSGPAGLYAQALFFSSPP (SEQ ID NO: 26); GPAGLYAQPLAQKLKSSALFKSSFP (SEQ ID NO: 27); GPAGLYAQPLAQKLKSSALFKSSFP (SEQ ID NO: 28); GPAGLYAQGPLAQKLKSSALFFSSPP (SEQ ID NO: 29); PLAQKLKSSGPAGLYAQALFKSSFP 69 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 (SEQ ID NO: 30); PLAQKLKSSGPAGLYAQALFKSSFP (SEQ ID NO: 31); PLAQKLKSSGPAGLYAQgpALFFSSPP (SEQ ID NO: 32); ISSGLLSGRSDNH (SEQ ID NO: 33); MPYDLYHP (SEQ ID NO: 34); VPLSLY (SEQ ID NO: 35); VLLVSEVL (SEQ ID NO: 37); TLLVSGNA (SEQ ID NO: 38); VVLFSEVL (SEQ ID NO: 39); GLLVSLGL (SEQ ID NO: 40); MPVAWEY (SEQ ID NO: 41); YYNEPY (SEQ ID NO: 42); FVTSW (SEQ ID NO: 43); HLFKSFPF (SEQ ID NO: 46); HKFKSKFP (SEQ ID NO: 47); KLLFHLFP (SEQ ID NO: 48); KKLFHIFP (SEQ ID NO: 49); ALFKSFPF (SEQ ID NO: 52); AKFRHLFP (SEQ ID NO: 53); ALLKSIFP (SEQ ID NO: 54); AKLRSKFP (SEQ ID NO: 55); PLYAKLKSSFP (SEQ ID NO: 56); PLAQKVKSSFP (SEQ ID NO: 57); or PLAQKLRSSFP (SEQ ID NO: 58).

[0256] Certain preferred linkers comprise the sequence VLLVSEVL (SEQ ID NO: 37), MPVAWEY (SEQ ID NO: 41), GPAGLYAQ (SEQ ID NO: 195), or GPAGLKGA (SEQ ID NO: 249).

[0257] In some embodiments, the linker comprises the sequence PMPVLFEYP (SEQ ID NO: 105), PMPAFWEYP (SEQ ID NO: 106), PMPVKWEKP (SEQ ID NO: 107), PMKAYVEDGKSP (SEQ ID NO: 108), PMPASLYAQP (SEQ ID NO: 112), PMPVFWPYP (SEQ ID NO: 113), PMPIKWEYP (SEQ ID NO: 114), PGLLVSLFHP (SEQ ID NO: 115).

[0258] The linkers disclosed herein can comprise one or more cleavage motifs or functional variants that are the same or different. The linkers can comprise 1, 2, 3, 4, 5, or more cleavage motifs or functional variants. Linkers comprising 30 amino acids can contain 2 cleavage motifs or functional variants, 3 cleavage motifs or functional variants or more. A “functional variant” of a linker retains the ability to be cleaved with high efficiency at a target site (e.g., a tumor microenvironment that expresses high levels of the protease) and are not cleaved or cleaved with low efficiency in the periphery (e.g., serum). For example, the functional variants retain at least about 50%, about 55%, about 60%, about 70%, about 80%, about 85%, about 95% or more of the cleavage efficiency of a linker comprising any one of SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220.

[0259] The linkers comprising more than one cleavage motif can be selected from SEQ ID NOs: 9-11, 14-16, 33-35, 39-43, 46-49, 52-55, 105-108, 112-115, or 195-201 and combinations thereof. Linkers comprising more than one cleavage motif can comprise the amino acids selected from SEQ ID NOs: 5, 17-32, 37-38, 56-58105-108, 112-115 or 202-220.

[0260] The linker can comprise both ALFKSSFP (SEQ ID NO: 198) and GPAGLYAQ (SEQ ID NO: 195). The linker can comprise two cleavage motifs that each have the sequence GPAGLYAQ (SEQ ID NO: 195). Alternatively, or additionally, the linker can comprise two cleavage motifs that each have the sequence ALFKSSFP (SEQ ID NO: 198). The linker can comprise a third cleavage motif that is the same or different. 70 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0261] In some embodiments, the linker comprises an amino acid sequence that is at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least 99% identical to SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 112-115, or 195-220 over the full length of SEQ ID NO: 5, 9-11, 14-35, 37-43, 46-49, 52-585, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220.

[0262] The disclosure also relates to functional variants of the linkers comprising SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220. The functional variants of the linkers comprising SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220 generally differ from SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220 by one or a few amino acids (including substitutions, deletions, insertions, or any combination thereof), and substantially retain their ability to be cleaved by a protease.

[0263] The functional variants can contain at least one or more amino acid substitutions, deletions, or insertions relative to the linkers comprising SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220. The functional variant can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid alterations comparted to the linkers comprising SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105- 108, 112-115, or 195-220. In some preferred embodiments, the functional variant differs from the linker comprising SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220 by less than 10, less, than 8, less than 5, less than 4, less than 3, less than 2, or one amino acid alterations, e.g., amino acid substitutions or deletions. In other embodiments, the functional variant may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions compared to SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220. The amino acid substitution can be a conservative substitution or a non- conservative substitution, but preferably is a conservative substitution. In some embodiments, the functional variants comprise an amino acid sequence that is at least about 90% identical to SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220.

[0264] In other embodiments, the functional variants of the linkers may comprise 1, 2, 3, 4, or 5 or more non-conservative amino acid substitutions compared to the linkers comprising SEQ ID NOs: 5, 9-11, 14- 35, 37-43, 46-49, 52-58, 105-108, 112-115, or 195-220. Non-conservative amino acid substitutions could be recognized by one of skill in the art. The functional variant of the linker preferably contains no more than 1, 2, 3, 4, or 5 amino acid deletions.

[0265] The amino acid sequences disclosed in the linkers can be described by the relative linear position in the linker with respect to the scissile bond. As will be well-understood by persons skilled in the art, linkers comprising 8 amino acid protease substrates (e.g., SEQ ID NOs: 195-201) contain amino acid at positions P4, P3, P2, P1, P1’, P2’, P3’, P4’, wherein the scissile bond is between P1 and P1’. For 71 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 example, amino acid positions for the linker comprising the sequence GPAGLYAQ (SEQ ID NO: 195) can be described as follows: G P A G L Y A Q P4 P3 P2 P1 P1’ P2’ P3’ P4’

[0266] Amino acids positions for the linker comprising the sequence ALFKSSFP (SEQ ID NO: 198) can be described as follows: A L F K S S F P P4 P3 P2 P1 P1’ P2’ P3’ P4’

[0267] Preferabnd P1’for SEQ ID NOs: 9-11, 14-16, 33-35, 39-43, 46-49, 52-58, 105-108, 112-115, or 195-201) are not substituted.

[0268] In embodiments, the linker comprises the sequence GPAGLYAQ (SEQ ID NO: 195) or ALFKSSFP (SEQ ID NO: 198) or a functional variant of SEQ ID NO: 195 or a function variant of SEQ ID NO: 198. As described herein, a functional variant of GPAGLYAQ (SEQ ID NO: 195) or ALFKSSFP (SEQ ID NO: 198) can comprise one or more amino acid substitutions, and substantially retain their ability to be cleaved by a protease. Specifically, the functional variants of GPAGLYAQ (SEQ ID NO: 195) are cleaved by MMP14, and the functional variants of ALFKSSFP (SEQ ID NO: 198) are cleaved by Capthepsin L (CTSL1). The functional variants also retain their ability to be cleaved with high efficiency at a target site (e.g., a tumor microenvironment that expresses high levels of the protease). For example, the functional variants of GPAGLYAQ (SEQ ID NO: 195) or ALFKSSFP (SEQ ID NO: 198) retain at least about 50%, about 55%, about 60%, about 70%, about 80%, about 85%, about 95% or more of the cleavage efficiency of a linker comprising amino acid sequence GPAGLYAQ (SEQ ID NO: 195) or ALFKSSFP (SEQ ID NO: 198), respectively.

[0269] Preferably, the functional variant of GPAGLYAQ (SEQ ID NO: 195) or ALFKSSFP (SEQ ID NO: 198) comprise no more than 1, 2, 3, 4, or 5 conservative amino acid substitutions compared to GPAGLYAQ (SEQ ID NO: 195) or ALFKSSFP (SEQ ID NO: 198). Preferably, the amino acids at position P1 and P1’ are not substituted. The amino acids at positions P1 and P1’ in SEQ ID NO: 195 are G and L, and the amino acids at positions P1 and P1’ in SEQ ID NO: 198 are K and S.

[0270] The functional variant of GPAGLYAQ (SEQ ID NO: 195) can preferably comprise one or more of the following: a) an arginine amino acid substitution at position P4, b) a leucine, valine, asparagine, or proline amino acid substitution at position P3, c) a asparagine amino acid substitution at position P2, d) a 72 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 histidine, asparagine, or glycine amino acid substitution at position P1, e) a asparagine, isoleucine, or leucine amino acid substitution at position P1’, f) a tyrosine or arginine amino acid substitution at position P2’, g) a glycine, arginine, or alanine amino acid substitution at position P3’, h) or a serine, glutamine, or lysine amino acid substitution at position P4’. The following amino acid substitutions are disfavored in functional variants of GPAGLYAQ (SEQ ID NO: 195): a) arginine or isoleucine at position P3, b) alanine at position P2, c) valine at position P1, d) arginine, glycine, asparagine, or threonine at position P1’, e) aspartic acid or glutamic acid at position P2’, f) isoleucine at position P3’, g) valine at position P4’. In some embodiments, the functional variant of GPAGLYAQ (SEQ ID NO: 195) does not comprise an amino acid substitution at position P1 and / or P1’.

[0271] The amino acid substitution of the functional variant of GPAGLYAQ (SEQ ID NO: 195) preferably comprises an amino acid substitution at position P4 and / or P4’. For example, the functional variant of GPAGLYAQ (SEQ ID NO: 195) can comprise a leucine at position P4, or serine, glutamine, lysine, or phenylalanine at position P4. Alternatively, or additionally, the functional variant of GPAGLYAQ (SEQ ID NO: 195) can comprise a glycine, phenylalanine, or a proline at position P4’.

[0272] In some embodiments, the amino acid substitutions at position P2 or P2’ of GPAGLYAQ (SEQ ID NO: 195) are not preferred.

[0273] In some embodiments, the functional variant of GPAGLYAQ (SEQ ID NO: 195) comprises the amino acid sequence selected from SEQ ID NOs: 221- 295. Specific functional variants of GPAGLYAQ (SEQ ID NO: 195) include GPLGLYAQ (SEQ ID NO: 259), and GPAGLKGA (SEQ ID NO: 249).

[0274] The functional variants of ALFKSSFP (SEQ ID NO: 198) preferably comprises hydrophobic amino acid substitutions. The functional variant of ALFKSSFP (SEQ ID NO: 198) can preferably comprise one or more of the following: (a) lysine, histidine, serine, glutamine, leucine, proline, or phenylalanine at position P4; (b) lysine, histidine, glycine, proline, asparagine, phenylalanine at position P3; (c) arginine, leucine, alanine, glutamine, or histatine at position P2; (d) phenylalanine, histidine, threonine, alanine, or glutamine at position P1; (e) histidine, leucine, lysine, alanine, isoleucine, arginine, phenylalanine, asparagine, glutamic acid, or glycine at position P1’, (f) phenylalanine, leucine, isoleucine, lysine, alanine, glutamine, or proline at position P2’; (g) phenylalanine, leucine, glycine, serine, valine, histidine, alanine, or asparagine at position P3’; and phenylalanine, histidine, glycine, alanine, serine, valine, glutamine, lysine, or leucine.

[0275] The inclusion of aspartic acid and / or glutamic acid in functional variants of SEQ ID NO: 448 are generally disfavored and avoided. The following amino acid substitutions are also disfavored in functional variants of LFKSSFP (SEQ ID NO: 448): (a) alanine, serine, or glutamic acid at position P3; (b) proline, threonine, glycine, or aspartic acid at position P2; (c) proline at position P1; (d) proline at 73 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 position P1’; (e) glycine at position P2’; (f) lysine or glutamic acid at position P3’; (g) aspartic acid at position P4’.

[0276] The amino acid substitution of the functional variant of LFKSSFP (SEQ ID NO: 448) preferably comprises an amino acid substitution at position P4 and / or P1. In some embodiments, an amino acid substitution of the functional variant of LFKSSFP (SEQ ID NO: 448) at position P4’ is not preferred.

[0277] In some embodiments, the functional variant of LFKSSFP (SEQ ID NO: 448) comprises the amino acid sequence selected from SEQ ID NOs: 296-374. Specific functional variants of LFKSSFP (SEQ ID NO: 448) include ALFFSSPP (SEQ ID NO: 199), ALFKSFPP (SEQ ID NO: 346), ALFKSLPP (SEQ ID NO: 347); ALFKHSPP (SEQ ID NO: 335); ALFKSIPP (SEQ ID NO: 348); ALFKSSLP (SEQ ID NO: 356); or SPFRSSRQ (SEQ ID NO: 297).

[0278] The linkers disclosed herein can form a stable prodrug under physiological conditions with the amino acid sequences (e.g., domains) that they link, while being capable of being cleaved by a protease. For example, the linker is stable (e.g., not cleaved or cleaved with low efficiency) in the circulation and cleaved with higher efficiency at a target site (i.e., a tumor microenvironment). Accordingly, inducible IL-10 prodrugs that include the linkers disclosed herein can, if desired, have a prolonged circulation half- life and / or lower biological activity in the circulation in comparison to the components of the inducible IL-10 prodrugs as separate molecular entities. Yet, when in the desired location (e.g., tumor microenvironment) the linkers can be efficiently cleaved to release the components that are joined together by the linker and restoring or nearly restoring the half-life and biological activity of the components as separate molecular entities.

[0279] The linker desirably remains stable in the circulation for at least 2 hours, at least 5, hours, at least 10 hours, at least 15 hours, at least 20 hours, at least 24 hours, at least 30 hours, at least 35 hours, at least 40 hours, at least 45 hours, at least 50 hours, at least 60 hours, at least 65 hours, at least 70 hours, at least 80 hours, at least 90 hours, or longer.

[0280] In some embodiments, the linker is cleaved by less than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 20%, 5%, or 1% in the circulation as compared to the target location. The linker is also stable in the absence of an enzyme capable of cleaving the linker. However, upon expose to a suitable enzyme (i.e., a protease), the linker is cleaved resulting in separation of the linked domain. F. Pharmaceutical Compositions

[0281] Described herein is a vector comprising the nucleotide sequence(s) encoding the inducible IL-10 prodrugs disclosed herein or a host cell transformed by this vector and at least one pharmaceutically acceptable carrier. 74 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0282] Provided herein are pharmaceutical formulations or compositions containing the inducible IL-10 prodrugs as described herein and a pharmaceutically acceptable carrier. Compositions comprising the inducible IL-10 prodrugs as described herein are suitable for administration in vitro or in vivo. The term "pharmaceutically acceptable carrier" includes, but is not limited to, any carrier that does not interfere with the effectiveness of the biological activity of the ingredients and that is not toxic to the subject to whom it is administered. Examples of suitable pharmaceutical carriers are well known in the art and include phosphate buffered saline solutions, water, emulsions, such as oil / water emulsions, various types of wetting agents, sterile solutions etc. Such carriers can be formulated by conventional methods and can be administered to the subject at a suitable dose. Preferably, the compositions are sterile. These compositions may also contain adjuvants such as preservative, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents.

[0283] Suitable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy, 21st Edition, David B. Troy, ed., Lippicott Williams & Wilkins (2005). Typically, an appropriate amount of a pharmaceutically acceptable salt is used in the formulation to render the formulation isotonic, although the formulate can be hypertonic or hypotonic if desired. Examples of the pharmaceutically acceptable carriers include, but are not limited to, sterile water, saline, buffered solutions like Ringer's solution, and dextrose solution. The pH of the solution is generally about 5 to about 8 or from about 7 to 7.5. Other carriers include sustained release preparations such as semipermeable matrices of solid hydrophobic polymers containing the immunogenic polypeptides. Matrices are in the form of shaped articles, e.g., films, lipid nanoparticles (LNPs), ferritin globular proteins, liposomes, or microparticles. Certain carriers may be more preferable depending upon, for instance, the route of administration and concentration of composition being administered. Carriers are those suitable for administration of the inducible IL-10 prodrugs or nucleic acid sequences encoding the inducible IL-10 prodrugs to humans or other subjects.

[0284] In some embodiments of the pharmaceutical compositions, the inducible IL-10 prodrugs described herein are encapsulated in nanoparticles. Exemplary nanoparticles include, but are not limited to, fullerenes, liquid crystals, liposome, quantum dots, superparamagnetic nanoparticles, dendrimers, or nanorods. The inducible IL-10 prodrug can be attached to liposomes. The inducible IL-10 prodrug can be conjugated to the surface of liposomes. The inducible IL-10 prodrug can be encapsulated within the shell of a liposome. The liposome can be a cationic liposome. In other embodiments of pharmaceutical compositions, the inducible IL-10 prodrug can be encapsulated in a solid lipid nanoparticle or a nanostructured lipid carrier. The lipid nanoparticle can consist of a lipid and emulsifier. 75 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0285] The inducible IL-10 prodrugs described herein are contemplated for use as a medicament. Administration is affected by different ways, e.g., by intravenous, intraperitoneal, subcutaneous, intramuscular, topical, intradermal, oral, parenteral, intra-articularly, intracavity, transdermal, intrahepatical, intracranial, inhalation, or intratumoral administration. In some embodiments, the route of administration depends on the kind of therapy and the kind of compound contained in the pharmaceutical composition. The dosage regimen will be determined by the attending physician and other clinical factors. Dosages for any one patient depends on many factors, including the patient's size, body surface area, age, sex, the particular compound to be administered, time and route of administration, the kind of therapy, general health and other drugs being administered concurrently. An "effective dose" refers to amounts of the active ingredient that are sufficient to affect the course and the severity of the disease, leading to the reduction or remission of such pathology and may be determined using known methods.

[0286] One or more nucleic acids sequences can encode the inducible IL-10 prodrugs disclosed herein, and these may be provided in a vector. Any suitable nucleic acid can be suitable for encoding the inducible IL-10 prodrugs disclosed herein. Exemplary nucleic acids include, but are not limited to, ribonucleic acids (RNAs), messenger RNA (mRNA), deoxyribonucleic acids (DNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic acids (LNAs, including LNA having a β- D-ribo configuration, a-LNA having an a-L-ribo configuration (a diastereomer of LNA), 2'-amino-LNA having a 2 '-amino functionalization, and 2'-amino- a-LNA having a 2'-amino functionalization), ethylene nucleic acids (ENA), cyclohexenyl nucleic acids (CeNA) or hybrids or combinations thereof.

[0287] One or more nucleic acids sequences can encode the inducible IL-10 prodrugs disclosed herein, and these may be composed of modified nucleosides or nucleotides known by one skilled in the art to reduce immune activation, evade exo- or endonucleases or improve pH stability. Exemplary modified nucleosides include, but are not limited to, pseudouridine (Ψ), N1-methyl-pseudouridine (N1-methyl- Ψ), 2-thiouridine (s2U), 5-fluoro-2’-deoxyuridine (FUDR), N1-methyladenosine (m1A), N6-methyladenosine (m6A), N6,2’-O-dimethyladenosine (m6Am), 7-methylguanosine, 8-oxo-7,8-dihydroguanosine (8-oxoG), 5-methylcytidine (m5C), N4-acetylcytidine (ac4C), 2’-deoxy-2’-fluoro-ribonucleosides, 2’-O- methoxyribonucleosides, N-ethylpiperidine-7-EAA triazole modified adenine, N-ethylpiperidine-6- triazole modified adenosine, 6-phenylpyrrolo-cytosine (PhpC), 2,4-difluorotoluyl-ribonucleoside (rF), N1(5-nitroindole) ribonucleoside, 2’-O-methyl-ribonucleosides (2’-OMe), 2’-O-methoxyethyl- ribonucleosides (2’-O-MOE), 2’-deoxy-2’-fluororibonucleosides (2’-F), 2’-deoxyarbino-2’- fluoronucleosides (2’-Ara-F), 2’-O-benzyl-2’-deoxyribonucleosides, 2’-O-methyl-4- 76 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 pyridinylribonucleosides (2’-O-CH2Py(4)), 5-methoxyuridine, 5-methyluridine, or hybrids or combinations thereof.

[0288] Exemplary nucleotides include, but are not limited to, phosphorothioate (Rp and / or Sp isomer, PS)-, phosphorodithioate (PS2)-, 5’-vinylphosphonate (5’-VP)-, 5’-methylphosphonate (5’-MP)-, methylphosphonate (MP)-, methoxypropylphosphonate (MOP)-, (S)-5’-C-methyl with phosphate- nucelotides, 5’,5’-(7-methylguanosine)phosphotriester, 5’,5’-(2’-O-methyl-7-guanosine)diphosphoro- phosphorothioate or hybrids or combinations thereof.

[0289] Described herein is a vector comprising the nucleotide sequence(s) encoding the inducible IL-10 prodrugs disclosed herein or a host cell transformed by this vector and at least one pharmaceutically acceptable carrier. Optionally, the inducible IL-10 prodrug or nucleic acid sequences encoding the inducible IL-10 prodrug are administered by a vector. There are a number of compositions and methods which can be used to deliver the nucleic acid molecules and / or polypeptides to cells, either in vitro or in vivo via, for example, expression vectors. These methods and compositions can largely be broken down into two classes: viral based delivery systems and non-viral based delivery systems. Such methods are well known in the art and readily adaptable for use with the compositions and methods described herein. Such compositions and methods can be used to transfect or transduce cells in vitro or in vivo, for example, to produce cell lines that express and preferably secrete the encoded chimeric polypeptide or to therapeutically deliver nucleic acids to a subject. The components of the inducible IL-10 prodrug disclosed herein are typically operably linked in frame to encode an inducible IL-10 prodrug.

[0290] As used herein, plasmid or viral vectors are agents that transport the disclosed nucleic acids into the cell without degradation and include a promoter yielding expression of the nucleic acid molecule and / or polypeptide in the cells into which it is delivered. Viral vectors are, for example, Adenovirus, Adeno-associated virus, herpes virus, Vaccinia virus, Polio virus, Sindbis, and other RNA viruses, including those viruses with the HIV backbone. Also preferred are any viral families which share the properties of these viruses which make them suitable for use as vectors. Retroviral vectors, in general and methods of making them are described by Coffin et al., Retroviruses, Cold Spring Harbor Laboratory Press (1997). The construction of replication-defective adenoviruses has been described (Berkner et al., J. Virol.61:1213-20 (1987); Massie et al., Mol. Cell. Biol.6:2872-83 (1986); Haj-Ahmad et al., J. Virol. 57:267-74 (1986); Davidson et al., J. Virol.61:1226-39 (1987); Zhang et al., BioTechniques 15:868-72 (1993)). The benefit and the use of these viruses as vectors is that they are limited in the extent to which they can spread to other cell types, since they can replicate within an initial infected cell, but are unable to form new infectious viral particles. Recombinant adenoviruses have been shown to achieve high efficiency after direct, in vivo delivery to airway epithelium, hepatocytes, vascular endothelium, CNS 77 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 parenchyma, and a number of other tissue sites. Other useful systems include, for example, replicating and host-restricted non-replicating vaccinia virus vectors.

[0291] The inducible IL-10 prodrugs and / or nucleic acid molecules disclosed herein can be delivered via virus like particles. Virus like particles (VLPs) consist of viral protein(s) derived from the structural proteins of a virus. Methods for making and using virus like particles are described in, for example, Garcea and Gissmann, Current Opinion in Biotechnology 15:513-7 (2004).

[0292] The inducible IL-10 prodrugs disclosed herein can be delivered by subviral dense bodies (DBs). DBs transport proteins into target cells by membrane fusion. Methods for making and using DBs are described in, for example, Pepperl-Klindworth et al., Gene Therapy 10:278-84 (2003). The provided polypeptides can be delivered by tegument aggregates. Methods for making and using tegument aggregates are described in International Publication No. WO 2006 / 110728.

[0293] Non-viral based delivery methods can include expression vectors comprising nucleic acid molecules and nucleic acid sequences encoding polypeptides, wherein the nucleic acids are operably linked to an expression control sequence. Suitable vector backbones include, for example, those routinely used in the art such as plasmids, artificial chromosomes, BACs, YACs, or PACs. Numerous vectors and expression systems are commercially available from such corporations as Novagen (Madison, Wis.), Clonetech (Pal Alto, Calif.), Stratagene (La Jolla, Calif.), and Invitrogen / Life Technologies (Carlsbad, Calif.). Vectors typically contain one or more regulatory regions. Regulatory regions include, without limitation, promoter sequences, enhancer sequences, response elements, protein recognition sites, inducible elements, protein binding sequences, 5′ and 3′ untranslated regions (UTRs), transcriptional start sites, termination sequences, polyadenylation sequences, and introns. Such vectors can also be used to make the inducible IL-10 prodrugs by expression in a suitable host cell, such as CHO cells.

[0294] Preferred promoters controlling transcription from vectors in mammalian host cells may be obtained from various sources, for example, the genomes of viruses such as polyoma, Simian Virus 40 (SV40), adenovirus, retroviruses, hepatitis B virus, and most preferably cytomegalovirus (CMV), or from heterologous mammalian promoters, e.g., β-actin promoter or EF1α promoter, or from hybrid or chimeric promoters (e.g., CMV promoter fused to the β-actin promoter). Of course, promoters from the host cell or related species are also useful herein.

[0295] Enhancer generally refers to a sequence of DNA that functions at no fixed distance from the transcription start site and can be either 5′ or 3′ to the transcription unit. Furthermore, enhancers can be within an intron as well as within the coding sequence itself. They are usually between 10 and 300 base pairs (bp) in length, and they function in cis. Enhancers usually function to increase transcription from nearby promoters. Enhancers can also contain response elements that mediate the regulation of 78 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 transcription. While many enhancer sequences are known from mammalian genes (globin, elastase, albumin, fetoprotein, and insulin), typically one will use an enhancer from a eukaryotic cell virus for general expression. Preferred examples are the SV40 enhancer on the late side of the replication origin, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.

[0296] The promoter and / or the enhancer can be inducible (e.g., chemically or physically regulated). A chemically regulated promoter and / or enhancer can, for example, be regulated by the presence of alcohol, tetracycline, a steroid, or a metal. A physically regulated promoter and / or enhancer can, for example, be regulated by environmental factors, such as temperature and light. Optionally, the promoter and / or enhancer region can act as a constitutive promoter and / or enhancer to maximize the expression of the region of the transcription unit to be transcribed. In certain vectors, the promoter and / or enhancer region can be active in a cell type specific manner. Optionally, in certain vectors, the promoter and / or enhancer region can be active in all eukaryotic cells, independent of cell type. Preferred promoters of this type are the CMV promoter, the SV40 promoter, the β-actin promoter, the EF1α promoter, and the retroviral long terminal repeat (LTR).

[0297] The vectors also can include, for example, origins of replication and / or markers. A marker gene can confer a selectable phenotype, e.g., antibiotic resistance, on a cell. The marker product is used to determine if the vector has been delivered to the cell and once delivered is being expressed. Examples of selectable markers for mammalian cells are dihydrofolate reductase (DHFR), thymidine kinase, neomycin, neomycin analog G418, hygromycin, puromycin, and blasticidin. When such selectable markers are successfully transferred into a mammalian host cell, the transformed mammalian host cell can survive if placed under selective pressure. Examples of other markers include, for example, the E. coli lacZ gene, green fluorescent protein (GFP), and luciferase. In addition, an expression vector can include a tag sequence designed to facilitate manipulation or detection (e.g., purification or localization) of the expressed polypeptide. Tag sequences, such as GFP, glutathione S-transferase (GST), polyhistidine, c-myc, hemagglutinin, or FLAG™ tag (Kodak; New Haven, Conn.) sequences typically are expressed as a fusion with the encoded polypeptide. Such tags can be inserted anywhere within the polypeptide including at either the carboxyl or amino terminus. G. Therapeutic Applications

[0298] Also provided herein, are methods and uses for the treatment of a disease, disorder or condition comprising administering to a subject in need thereof an inducible IL-10 prodrug or nucleic acid encoding the same as described herein. Diseases, disorders, or conditions include, but are not limited to, cancer, 79 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 inflammatory disease, an immunological disorder, autoimmune disease, infectious disease (i.e., bacterial, viral, or parasitic disease). Preferably, the disease, disorder, or condition is an inflammatory disease or autoimmune disease. The inducible IL-10 prodrugs, nucleic acids, vectors and compositions disclosed herein can be used in therapy. The inducible IL-10 prodrugs, nucleic acids, vectors and compositions disclosed herein can be used as a medicament. The IL-10 prodrugs, nucleic acids, vectors and compositions disclosed herein can also be used in methods of treating specific diseases, disorders, or conditions.

[0299] The methods and compositions disclosed herein can be used to treat any suitable inflammatory disease, in particular long-term or chronic inflammatory diseases, such as inflammatory bowel diseases. In some embodiments, the methods and compositions disclosed herein can be used to treat inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), psoriasis, celiac disease, ankylosing spondylitis, gout, Sjogren’s syndrome, systemic lupus erythematosus, rheumatoid arthritis, myositis, vasculitis (Behçet’s disease), psoriatic arthritis, rosacea, primary biliary cholangitis, osteoarthritis, pemphigoid, pemphigus, myasthenia gravis, lupus nephritis, asthma, such as allergic asthma, eosinophilic asthma, or non-allergic asthma, bronchitis, atopic dermatitis, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic obstructive pulmonary disease (COPD), sarcoidosis, graft versus host disease (GVHD), periodontitis, endometriosis, conjunctivitis, uveitis, bursitis, seroma, multiple sclerosis (MS), diabetes mellitus, pancreatitis, encephalomyelitis, encephalitis, sepsis, cytokine storm, eczema, allergic rhinitis, eosinophilic esophagitis, neuromyelitis optica, idiopathic inflammatory demyelinating disease, diffuse myelinoclastic sclerosis, Guillain-Barré syndrome, Alzheimer’s disease, cirrhosis, nephritis, aplastic anemia, autoimmune thyroiditis, thyroiditis, fibromyalgia, myocarditis, endocarditis, pericarditis, pelvic inflammatory disease, hepatitis, glomerulonephritis, ischemia-reperfusion injury, fatty liver disease, obesity, sinusitis, scleroderma, Graves’ disease, tonsillitis, appendicitis, osteoporosis, Hashimoto’s thyroiditis, Addison’s disease, adult Still’s disease, agammaglobulinemia, amyloidosis, anti-glomerular basement membrane disease, autoimmune angioedema, autoimmune dysautonomia, autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear disease, autoimmune myocarditis, autoimmune oophoritis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal and neuronal neuropathy, Castleman disease, chronic inflammatory demyelinating polyneuropathy, chronic recurrent multifocal osteomyelitis, Churg-Strauss syndrome, eosinophilic granulomatosis, Cogan’s syndrome, CREST syndrome, dermatitis herpetiformis, discoid lupus, Dressler syndrome, eosinophilic fasciitis, erythema nodosum, Evans syndrome, idiopathic pulmonary fibrosis, giant cell arteritis, giant cell myocarditis, granulomatosis with polyangiitis, hemolytic anemia, Henoch-Schönlein purpura, hidradenitis suppurativa, IgA nephropathy, IgG4-related sclerosing disease, immune 80 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 thrombocytopenic purpura, inclusion body myositis, interstitial cystitis, Lewy body dementia, juvenile arthritis, juvenile myositis, Kawasaki disease, Lambert-Eaton syndrome, lichen planus, lichen sclerosus, chronic Lyme disease, microscopic polyangiitis, Mooren’s ulcer, Mucha-Habermann disease, neonatal lupus, palindromic rheumatism, paraneoplastic cerebellar degeneration, chronic intermediate uveitis, polyarteritis nodosa, Autoimmune polyendocrine syndromes (types I, II, III), polymyalgia rheumatica, Takatsuki disease, primary sclerosing cholangitis, progesterone dermatitis, pyoderma gangrenosum, reactive arthritis, complex regional pain syndrome, polychondritis, retroperitoneal fibrosis, scleritis, scleroderma, still person syndrome, autoimmune endotheliopathy, Takayasu’s arthritis, Alopecia areata, Wiskott-Aldrich syndrome, atopic dermatitis, vitiligo, vasculitis, IPEX syndrome, or transverse myelitis.

[0300] In certain embodiments, the methods and compositions disclosed herein can be used to treat chronic or episodic flares of inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), psoriasis, celiac disease, ankylosing spondylitis, gout, Sjogren’s syndrome, systemic lupus erythematosus, rheumatoid arthritis, myositis, vasculitis (Behçet’s disease), psoriatic arthritis, rosacea, primary biliary cholangitis, osteoarthritis, pemphigoid, pemphigus, myasthenia gravis, lupus nephritis, asthma, such as allergic asthma, eosinophilic asthma, non-allergic asthma, bronchitis, atopic dermatitis, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic obstructive pulmonary disease (COPD), sarcoidosis, graft versus host disease (GVHD), periodontitis, endometriosis, conjunctivitis, uveitis, bursitis, seroma, multiple sclerosis (MS), diabetes mellitus, pancreatitis, encephalomyelitis, encephalitis, sepsis, cytokine storm, eczema, allergic rhinitis, eosinophilic esophagitis, neuromyelitis optica, idiopathic inflammatory demyelinating disease, diffuse myelinoclastic sclerosis, Guillain-Barré syndrome, Alzheimer’s disease, cirrhosis, nephritis, aplastic anemia, autoimmune thyroiditis, thyroiditis, fibromyalgia, myocarditis, endocarditis, pericarditis, pelvic inflammatory disease, hepatitis, glomerulonephritis, ischemia-reperfusion injury, fatty liver disease, obesity, sinusitis, scleroderma, Graves’ disease, tonsillitis, appendicitis, osteoporosis, Hashimoto’s thyroiditis, Addison’s disease, adult Still’s disease, agammaglobulinemia, amyloidosis, anti-glomerular basement membrane disease, autoimmune angioedema, autoimmune dysautonomia, autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear disease, autoimmune myocarditis, autoimmune oophoritis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal and neuronal neuropathy, Castleman disease, chronic inflammatory demyelinating polyneuropathy, chronic recurrent multifocal osteomyelitis, Churg-Strauss syndrome, eosinophilic granulomatosis, Cogan’s syndrome, CREST syndrome, dermatitis herpetiformis, discoid lupus, Dressler syndrome, eosinophilic fasciitis, erythema nodosum, Evans syndrome, idiopathic pulmonary fibrosis, giant cell arteritis, giant cell myocarditis, granulomatosis with polyangiitis, hemolytic anemia, Henoch-Schönlein purpura, hidradenitis suppurativa, IgA nephropathy, 81 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 IgG4-related sclerosing disease, immune thrombocytopenic purpura, inclusion body myositis, interstitial cystitis, Lewy body dementia, juvenile arthritis, juvenile myositis, Kawasaki disease, Lambert-Eaton syndrome, lichen planus, lichen sclerosus, chronic Lyme disease, microscopic polyangiitis, Mooren’s ulcer, Mucha-Habermann disease, neonatal lupus, palindromic rheumatism, paraneoplastic cerebellar degeneration, chronic intermediate uveitis, polyarteritis nodosa, Autoimmune polyendocrine syndromes (types I, II, III), polymyalgia rheumatica, Takatsuki disease, primary sclerosing cholangitis, progesterone dermatitis, pyoderma gangrenosum, reactive arthritis, complex regional pain syndrome, polychondritis, retroperitoneal fibrosis, scleritis, scleroderma, still person syndrome, autoimmune endotheliopathy, Takayasu’s arthritis, or transverse myelitis.

[0301] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat rheumatoid arthritis, asthma, COPD, diabetes, Alzheimer’s disease, cirrhosis, inflammatory bowel diseases (e.g., Crohn’s disease, ulcerative colitis) or sepsis.

[0302] Preferably, the methods and compositions disclosed herein are used to treat rheumatoid arthritis, allergic asthma, COPD, diabetes, Alzheimer’s disease, cirrhosis, inflammatory bowel diseases (e.g., Crohn’s disease, ulcerative colitis) or sepsis.

[0303] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). As an example, the methods and compositions disclosed herein can be used to treat inflammatory bowel disease in subjects diagnosed with ulcerative colitis, such as relapsing or flaring ulcerative colitis. In another example, the methods and compositions disclosed herein can be used to treat proctitis, left-sided colitis or extensive (pan) colitis. As another example, the methods and compositions disclosed herein can be used for the adjuvant treatment of subjects with ulcerative colitis following or prior to proctocolectomy with ileal pouch-anal anastomosis or proctocolectomy with end ileostomy. As a further example, the methods and compositions disclosed herein can be used as maintenance treatment of ulcerative colitis in remission or prevention of ulcerative colitis relapse. As another example, the methods disclosed herein can be used for treatment of ulcerative colitis subjects following Mayo scoring or disease activity indexing or other disease scoring methods for disease severity or diagnosis of disease. As another example, the methods and compositions disclosed herein can be used in combination or concurrently with other treatments for ulcerative colitis well known to one skilled in the art including, but not limited to, immunomodulators (e.g., azathioprine, 6- mercaptopurine), corticosteroids (e.g., methylprednisolone, budesonide, prednisone), aminosalicylate anti-inflammatories (e.g., sulfasalazine, olsalazine, basalazide, mesalamine), biologics (e.g., adalimumab certolizumab, infliximab; natalizumab, vedolizumab; ustekinumab) or hybrids or combinations of the treatments thereof. 82 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0304] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). As an example, the methods and compositions disclosed herein can be used to treat inflammatory bowel disease in subjects diagnosed with Crohn’s disease. In another example, the methods and compositions disclosed herein can be used to treat ileoceacal Crohn’s disease, Crohn’s colitis, ileocolitis, gastroduodenal Crohn’s disease, ileitis, jejunoileitis or fistulating Crohn’s disease. As another example, the methods and compositions disclosed herein can be used for the adjuvant treatment of subjects with Crohn’s disease following or prior to ostomy, ileostomy, colostomy, colectomy, strictureplasty. As a further example, the methods and compositions disclosed herein can be used as maintenance treatment of Crohn’s disease in remission or prevention of Crohn’s disease relapse. As another example, the methods disclosed herein can be used for treatment of Crohn’s disease subjects following Crohn’s disease Endoscopic Index of Severity (CDEIS), Simple Endoscopic Score for Crohn’s disease (CES-CD), or other disease scoring for disease severity or diagnosis of disease. As another example, the methods and compositions disclosed herein can be used in combination or concurrently with other treatments for Crohn’s disease well known to one skilled in the art including, but not limited to, immunomodulators (e.g., azathioprine, 6-mercaptopurine), corticosteroids (e.g., methylprednisolone, budesonide, prednisone), aminosalicylate anti-inflammatories (e.g., sulfasalazine, olsalazine, basalazide, mesalamine), biologics (e.g., adalimumab certolizumab, infliximab, natalizumab, vedolizumab, ustekinumab) or hybrids or combinations of the treatments thereof.

[0305] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat celiac (coeliac) disease. As an example, the methods and compositions disclosed herein can be used to treat celiac disease in subjects with positive test result for HLA-DQ2 and / or HLA-DQ8 antigens, anti- endomysial (EMA) IgA, anti-transglutaminase antibodies, endoscopy / gastroscopy, biopsy, anti-reticulin (ARA) antibodies, anti-gliadin antibodies, or combinations thereof. As another example, the methods and compositions disclosed herein can be used to treat celiac disease in patients prior to, concurrent with or after cessation of gluten-free diet regimens. As another example, the methods and compositions disclosed herein can be used in combination with other therapies for celiac disease patients known to those skilled in the art to treat symptoms, achieve or maintain remission, or treat refractory disease including, but not limited to, antibiotics (e.g., dapsone), immunomodulators (e.g., azathioprine, 6-mercaptopurine), corticosteroids (e.g., methylprednisolone, budesonide, prednisone), hybrids, or combinations thereof.

[0306] In certain embodiments, the methods and compositions disclosed herein are used to treat chronic obstructive pulmonary disease (COPD). As an example, the methods and compositions disclosed herein can be used to treat COPD in subjects diagnosed with disease by pulmonary function tests, chest X-ray, CT scan, arterial blood gas analysis, test positive for alpha-1-antitrypsin (AAT) deficiency, or a hybrid or 83 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 combination thereof. As another example, the methods and compositions disclosed herein can be used to treat any form of COPD including chronic bronchitis and emphysema. As another example, the methods and compositions disclosed herein can be used to treat stable COPD, early diagnosis COPD, progressive COPD, frequent acute exacerbations of COPD (AECOPD), late-stage COPD, or any combination or hybrid therein. As another example, the methods and compositions disclosed herein can be used to treat patients with COPD undergoing surgery such as bullectomy, lung volume reduction surgery (LVRS) and lung transplantation; with the treatment being used prior to or following surgical intervention. As another example, the methods and compositions disclosed herein can be used in combination with other therapies for patients of COPD known to those skilled in the art including, but not limited to, short-acting bronchodilators (e.g., albuterol, ipratropium, levabuterol), long-acting bronchodilators (e.g., aclidinium, arformoterol, formoterol, indacaterol, tiotropium, salmeterol, umeclidinium), inhaled steroids (e.g., fluticasone, budesonide), phosphodiesterase-4 inhibitor (e.g., roflumilast), xanthines (e.g., theophylline), hybrids or combinations thereof.

[0307] In certain embodiments, the methods and compositions disclosed herein are used to treat rheumatoid arthritis. As an example, the methods and compositions disclosed herein can be used to treat rheumatoid arthritis in subjects either testing positive or negative for rheumatoid factor (RF). As another example, the methods and compositions disclosed herein can be used to treat rheumatoid arthritis in subjects diagnosed by Visual Analog Scale (VAS), Disease Activity Score at 28 joints (DAS28), other scale known to one skilled in the art, or a combination or hybrid thereof. As another example, the methods and compositions disclosed herein can be used to treat rheumatoid arthritis in subjects who have tested positive for elevated erythrocyte sedimentation rate (ESR, >27 mm / h), elevated C-reactive protein (CRP, >10.0 mg / L), elevated anti-cyclic citrullinated peptide (anti-CCP) antibodies, a hybrid, or combination thereof. As another example, the methods and compositions disclosed herein can be used in combination with treatments for rheumatoid arthritis known to one skilled in the art including, but not limited to, NSAIDs (e.g., ibuprofen, naproxen), corticosteroids (e.g., prednisone, methylprednisolone) immunomodulatory agents (e.g., methotrexate, leflunomide, baricitinib, tofacitinib, upadacitinib), biologics (e.g., adalimumab, etanercept; rituximab; tocilizumab; anakinra; abatacept), hybrids, or combinations thereof.

[0308] In certain embodiments, the methods and compositions disclosed herein are used to treat asthma. As an example, the methods and compositions disclosed herein can be used to treat all stages of asthma, including but not limited to, intermittent asthma, mild persistent asthma, moderate persistent asthma, severe persistent asthma or hybrids or combinations thereof. As another example, the methods and compositions disclosed herein can be used to treat asthma caused by common or uncommon allergens, 84 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 including but not limited to, dander, pollen, mold, dust mites, cockroaches, hybrids or combinations thereof. As another example, the methods and compositions disclosed herein can be used to treat asthma diagnosed by a positive test via spirometry, exhaled nitric oxide test, bronchoprovocation test, skin allergy test, blood IgE test, hybrids or combinations thereof. As another example, the methods and compositions disclosed herein can be used to treat asthma in remission, to maintain asymptomatic asthma or to maintain a low asthma severity stage. As another example, the methods and compositions disclosed herein can be used to treat a patient with asthma during an acute asthmatic attack, asthmatic flare-up, or as an asthmatic rescue agent. As another example, the methods and compositions disclosed herein can be used to treat a patient with asthma that has a disease resistant to first- or second-line treatments. As another example, the methods and compositions disclosed herein can be used in combination with treatments for asthma known to one skilled in the art including, but not limited to, inhaled short-acting beta-2-agonists (e.g., albuterol, levalbuterol, ipratropium bromide), inhaled steroids (e.g., fluticasone, budesonide, dexamethasone), inhaled long-acting beta-2-agonists (e.g., formoterol, salmeterol), leukotriene-modifying agents (e.g., Zafirlukast, Zileuton), xanthines (e.g., theophylline), beta-2-agonist tablets (levalbuterol, metaproterenol), IL-4 antagonist (e.g., reslizumab), steroid tablets (e.g., prednisone, beclomethasone dipropionate), IL-5 antagonist (e.g., mepolizumab), IL-5 receptor antagonist (e.g., benralizumab), hybrids, or combinations thereof.

[0309] In some embodiments, provided herein is a method of attenuating a disproportionate autoimmune or inflammatory response in a subject in need thereof by administering an effective amount of an inducible IL-10 prodrug provided herein, or nucleic acid encoding the same, to the subject. The attenuated immune response may prevent, delay, or treat the onset of an autoimmune or an inflammatory disease. Without being bound by theory, the inducible IL-10 prodrug attenuates the immune response by suppressing the innate and adaptive immunities. In some embodiments, the methods described herein suppress the activation or priming of Natural Killer Cells and T lymphocytes. In some embodiments, the inducible IL-10 prodrug provided herein, can reduce inflammatory cytokine release (e.g., IL-17a, IL-17f, IFNγ) from Natural Killer cells as well as CD4+ and CD8+ T cells. In some embodiments the inducible IL-10 prodrug can suppress the inflammatory activity or activation of myeloid cells including, but not limited to, monocytes, neutrophils, mononuclear phagocytes, macrophages and dendritic cells. In some embodiments the inducible IL-10 prodrug can suppress the phagocytosis, antigen presentation and T cell priming by dendritic cells, monocytes, macrophages and mononuclear phagocytes. In some embodiments the inducible IL-10 prodrug can suppress the cytokine (IL-6, IL-12, TNFα, IL-23) production by dendritic cells, monocytes, macrophages and mononuclear phagocytes. In some embodiments the inducible IL-10 85 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 prodrug can induce interleukin-1 receptor antagonist (IL-1RN) production by dendritic cells, monocytes, macrophages and mononuclear phagocytes.

[0310] This also disclosure relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with immunomodulator therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any desired immunomodulator therapeutic.

[0311] Exemplary immunomodulator therapeutics that can be combined with inducible IL-10 prodrugs or nucleic acids encoding the same include, but are not limited to, azathioprine, 6-mercaptopurine, methotrexate, tacrolimus, ozanimod, tofacitinib, udapacitinib, cladribine, montelukast, baricitinib, leflunomide, auranofin, misoprostol, ketoprofen, penicillamine, cyclophosphamide, a hybrid treatment, or combination thereof.

[0312] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with an immunomodulator therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination an immunomodulator therapeutic.

[0313] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with steroidal therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any desired steroidal therapeutic.

[0314] Exemplary steroidal therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, methylprednisolone, budesonide, prednisone, hydrocortisone, hydrocortisone acetate, prednisolone, beclomethasone dipropionate, dexamethasone, beclomethasone dipropionate, clobetasone butyrate, fluticasone, triamcinolone, cortisone, mometasone, ciclesonide, hybrids, or combination thereof.

[0315] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with a steroidal therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination a steroidal therapeutic.

[0316] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with non-steroidal anti-inflammatory 86 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 drugs (NSAIDs). The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any desired non-steroidal anti-inflammatory drug (NSAID).

[0317] Exemplary non-steroidal anti-inflammatory drugs (NSAIDs) that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, sulfasalazine, olsalazine, basalazide, mesalamine, aspirin, acetaminophen, ibuprofen, naproxen, celecoxib, meloxicam, diclofenac, nabumetone, etodolac, indomethacin, oxaprozin, piroxicam, salsalate, sulindac, methyl salicylate, diflunisal, fenoprofen, flurbiprofen, meclofenamate, tolmetin, or hybrids, or combination thereof.

[0318] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with a non-steroidal anti-inflammatory drugs (NSAIDs) as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination a non-steroidal anti- inflammatory drug (NSAID).

[0319] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with antimicrobial therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any antimicrobial therapeutic.

[0320] Exemplary antimicrobial therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, ciprofloxacin, metronidazole, dapsone, dapsone, sulfapyridine, sulfasalazine, or hybrids, or combination thereof.

[0321] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with an antimicrobial therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination an antimicrobial therapeutic.

[0322] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with biologic therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any biologic therapeutic.

[0323] Exemplary biologic therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, adalimumab, certolizumab pegol, inflixaimab, natalizumab, vedolizumab, ustekinumab, golimumab, alemtuzumab, PRV-015, AVX-176, 87 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 etanercept, rituximab, tocilizumab, anakinra, abatacept, sarilumab, reslizumab, mepolizumab, benralizumab, tezepelumab, dupilumab, omalizumab, or hybrids, or combination thereof.

[0324] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with a biologic therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination a biologic therapeutic.

[0325] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with enzyme therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any enzyme therapeutic.

[0326] Exemplary enzyme therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, latiglutenase, endoprotease B, prolyl endopeptidase, or hybrids, or combination thereof.

[0327] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with an enzyme therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination an enzyme therapeutic.

[0328] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with long- or short-acting bronchodilator therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any long- or short-acting bronchodilator therapeutic.

[0329] Exemplary long- or short-acting bronchodilator therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, albuterol, levabuterol, aclidinium, arformoterol, formoterol, indacaterol, olodaterol, revefenacin, tiotropium, salmeterol, umeclidinium, vilanterol, terbutaline, levalbuterol, metaproterenol, epinephrine, or hybrids, or combination thereof.

[0330] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with a long- or short-acting bronchodilator therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that 88 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination a long- or short- acting bronchodilator therapeutics.

[0331] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with xanthine therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any xanthine therapeutic.

[0332] Exemplary xanthine therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, theophylline, caffeine, hybrids, or combination thereof.

[0333] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with a xanthine therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination a xanthine therapeutic.

[0334] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with phosphodiesterase-4 inhibitor therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any phosphodiesterase-4 inhibitor therapeutic.

[0335] Exemplary phosphodiesterase-4 inhibitor therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, roflumilast, hybrids, or combination thereof.

[0336] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with a phosphodiesterase-4 inhibitor therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination a phosphodiesterase-4 inhibitor therapeutic.

[0337] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with leukotriene-modifying therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any leukotriene-modifying therapeutic. 89 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0338] Exemplary leukotriene-modifying therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, Zafirlukast, zileuton, montelukast, hybrids, or combination thereof.

[0339] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with a leukotriene-modifying therapeutic as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination a leukotriene-modifying therapeutic.

[0340] This disclosure also relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with other autoimmune or inflammatory disease modifying therapeutics. The inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any other autoimmune or inflammatory disease modifying therapeutics.

[0341] Exemplary other autoimmune or inflammatory disease modifying therapeutics that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, guaifenesin, dyphylline, cromolyn, hydroxychloroquine, glycopyrrolate, hybrids, or combination thereof.

[0342] This disclosure further relates to methods for treating inflammatory or autoimmune diseases using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with other autoimmune or inflammatory disease modifying therapeutics as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination other autoimmune or inflammatory disease modifying therapeutics.

[0343] The methods and compositions disclosed herein can be used to treat any suitable cancer. For example, the methods and compositions disclosed herein can be used to treat acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, brain tumor, bile duct cancer, bladder cancer, bone cancer, breast cancer, bronchial tumor, carcinoma of unknown primary origin, cardiac tumor, cervical cancer, chordoma, colon cancer, colorectal cancer, craniopharyngioma, ductal carcinoma, embryonal tumor, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, fibrous histiocytoma, Ewing sarcoma, eye cancer, germ cell tumor, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, gestational trophoblastic disease, glioma, head and neck cancer, hepatocellular cancer, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, 90 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 islet cell tumor, Kaposi sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ, lung cancer, macroglobulinemia, malignant fibrous histiocytoma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer with occult primary, midline tract carcinoma involving NUT gene, mouth cancer, multiple endocrine neoplasia syndrome, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, nasal cavity and par nasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytomas, pituitary tumor, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell cancer, renal pelvis and ureter cancer, retinoblastoma, rhabdoid tumor, salivary gland cancer, Sezary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, spinal cord tumor, stomach cancer, T-cell lymphoma, teratoid tumor, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, and Wilms tumor.

[0344] In certain embodiments, the methods and compositions disclosed herein are used to treat melanoma, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), head and neck squamous cell cancer (HNSCC), classical Hodgkin lymphoma (cHL), primary mediastinal large B cell lymphoma (PMBCL), urothelial carcinoma, microsatellite instability high or mismatch repair deficient cancer, microsatellite instability high or mismatch repair deficient colorectal cancer, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma (HCC), merkel cell carcinoma (MCC), renal cell carcinoma (RCC), endometrial carcinoma, tumor mutational burden high cancer, cutaneous squamous cell carcinoma (cSCC), triple negative breast cancer (TNBC), urothelial carcinoma, colorectal cancer or oesophageal carcinoma.

[0345] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat Merkel Cell Carcinoma (MCC), Urothelial Carcinoma (UC), Renal Cell Carcinoma (RCC), non- small cell lung cancer (NSCLC), small cell lung cancer (SCLC), triple negative breast cancer (TNBC), endometrial cancer, cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), melanoma, malignant pleural mesothelioma, classical Hodgkin lymphoma (cHL), squamous cell carcinoma of the head and neck (SCCHN), hepatocellular carcinoma (HCC), esophageal squamous cell carcinoma (ESCC), non-squamous non-small cell lung cancer, or nasopharyngeal carcinoma (NPC).

[0346] Preferably, the methods and compositions disclosed herein are used to treat colon cancer, lung cancer, melanoma, renal cell carcinoma, or breast cancer. 91 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0347] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat melanoma. As an example, the methods and compositions disclosed herein can be used to treat melanoma in subjects with unresectable or metastatic melanoma. As another example, the methods and compositions disclosed herein can be used for the adjuvant treatment of subjects with melanoma with involvement of lymph node(s) following complete resection.

[0348] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat non-small cell lung cancer (NSCLC). As an example, the methods and compositions disclosed herein can be used to treat NSCLC in subjects with NSCLC expressing PD-L1 (e.g., Tumor Proportion Score (TPS) ≥1%) as determined by an FDA-approved test, with no EGFR or ALK genomic tumor aberrations, and is: stage III where subjects are not candidates for surgical resection or definitive chemoradiation, or metastatic. As another example, the methods and compositions disclosed herein can be used to treat NSCLC in patients with metastatic NSCLC whose tumors express PD-L1 (TPS ≥1%) as determined by an FDA-approved test, with disease progression on or after platinum-containing chemotherapy. As another example, the methods and compositions disclosed herein can be used in combination with pemetrexed and platinum chemotherapy, as first-line treatment of patients with metastatic nonsquamous NSCLC, with no EGFR or ALK genomic tumor aberrations. As another example, the methods and compositions disclosed herein can be used in combination with carboplatin and either paclitaxel or paclitaxel protein- bound, as first-line treatment of patients with metastatic squamous NSCLC.

[0349] In certain embodiments, the methods and compositions disclosed herein are used to treat SCLC. As an example, the methods and compositions disclosed herein can be used to treat SCLC in subjects with metastatic SCLC with disease progression on or after platinum-based chemotherapy and at least one other prior line of therapy.

[0350] In certain embodiments, the methods and compositions disclosed herein are used to treat HNSCC. As an example, the methods and compositions disclosed herein can be used to treat HNSCC in subjects with metastatic or with unresectable, recurrent HNSCC whose tumors express PD-L1 (e.g., Combined Positive Score (CPS) ≥1) as determined by an FDA-approved test. As another example, the methods and compositions disclosed herein can be used to treat HNSCC in subjects with recurrent or metastatic HNSCC with disease progression on or after platinum-containing chemotherapy. As another example, the methods and compositions disclosed herein can be used in combination with platinum and fluorouracil for the first-line treatment of patients with metastatic or with unresectable, recurrent HNSCC.

[0351] In certain embodiments, the methods and compositions disclosed herein are used to treat cHL. As an example, the methods and compositions disclosed herein can be used to treat cHL in subjects with relapsed or refractory cHL. As another example, the methods and compositions disclosed herein can be 92 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 used to treat cHL in pediatric subjects with refractory cHL, or cHL that has relapsed after 2 or more lines of therapy.

[0352] In certain embodiments, the methods and compositions disclosed herein are used to treat urothelial carcinoma. As an example, the methods and compositions disclosed herein can be used to treat urothelial carcinoma in subjects with locally advanced or metastatic urothelial carcinoma who are not eligible for cisplatin-containing chemotherapy and whose tumors express PD-L1 (e.g., Combined Positive Score (CPS) ≥10) as determined by an FDA-approved test, or in subjects who are not eligible for any platinum-containing chemotherapy regardless of PD-L1 status. As another example, the methods and compositions disclosed herein can be used to treat urothelial carcinoma in subjects with locally advanced or metastatic urothelial carcinoma who have disease progression during or following platinum-containing chemotherapy or within 12 months of neoadjuvant or adjuvant treatment with platinum-containing chemotherapy. As another example, the methods and compositions disclosed herein can be used to treat urothelial carcinoma in subjects with Bacillus Calmette-Guerin (BCG)-unresponsive, high-risk, non- muscle invasive bladder cancer (NMIBC) with carcinoma in situ (CIS) with or without papillary tumors who are ineligible for or have elected not to undergo cystectomy.

[0353] In certain embodiments, the methods and compositions disclosed herein are used to treat Microsatellite Instability-High (MSI-H) or Mismatch Repair Deficient (dMMR) Cancer. As an example, the methods and compositions disclosed herein can be used to treat MSI-H or dMMR cancer in subjects with unresectable or metastatic MSI-H or dMMR cancer wherein the solid tumors have progressed following prior treatment and the subject has no satisfactory alternative treatment options, or wherein the colorectal cancer has progressed following treatment with a fluoropyrimidine, oxaliplatin, and irinotecan.

[0354] In certain embodiments, the methods and compositions disclosed herein are used to treat Microsatellite Instability-High (MSI-H) or Mismatch Repair Deficient (dMMR) Colorectal Cancer. As an example, the methods and compositions disclosed herein can be used to treat MSI-H or dMMR colorectal cancer in subjects with unresectable or metastatic MSI-H or dMMR colorectal cancer.

[0355] In certain embodiments, the methods and compositions disclosed herein are used to treat gastric cancer. As an example, the methods and compositions disclosed herein can be used to treat gastric cancer in subjects with recurrent locally advanced or metastatic gastric or gastroesophageal junction adenocarcinoma whose tumors express PD-L1 (e.g., Combined Positive Score (CPS) ≥1) as determined by an FDA-approved test, with disease progression on or after 2 or more prior lines of therapy including fluoropyrimidine- and platinum-containing chemotherapy and if appropriate, HER2 / neu-targeted therapy.

[0356] In certain embodiments, the methods and compositions disclosed herein are used to treat esophageal cancer. As an example, the methods and compositions disclosed herein can be used to treat 93 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 esophageal cancer in subjects with locally advanced or metastatic esophageal or gastroesophageal junction (GEJ) (e.g., tumors with epicenter 1 to 5 centimeters above the GEJ) carcinoma that is not amenable to surgical resection or definitive chemoradiation, in combination with platinum- and fluoropyrimidine-based chemotherapy. As another example, the methods and compositions disclosed herein can be used to treat esophageal cancer in subjects with locally advanced or metastatic esophageal or gastroesophageal junction (GEJ) (e.g., tumors with epicenter 1 to 5 centimeters above the GEJ) carcinoma that is not amenable to surgical resection or definitive chemoradiation, after one or more prior lines of systemic therapy for patients with tumors of squamous cell histology that express PD-L1 (CPS ≥10) as determined by an FDA-approved test.

[0357] In certain embodiments, the methods and compositions disclosed herein are used to treat cervical cancer. As an example, the methods and compositions disclosed herein can be used to treat cervical cancer in subjects with recurrent or metastatic cervical cancer with disease progression on or after chemotherapy whose tumors express PD-L1 (e.g., Combined Positive Score (CPS) ≥1) as determined by an FDA-approved test.

[0358] In certain embodiments, the methods and compositions disclosed herein are used to treat HCC. As an example, the methods and compositions disclosed herein can be used to treat HCC in subjects who have been previously treated with sorafenib.

[0359] In certain embodiments, the methods and compositions disclosed herein are used to treat MCC. As an example, the methods and compositions disclosed herein can be used to treat MCC in subjects with recurrent locally advanced or metastatic MCC.

[0360] In certain embodiments, the methods and compositions disclosed herein are used to treat RCC. As an example, the methods and compositions disclosed herein can be used in combination with axitinib, for the first-line treatment of patients with advanced RCC.

[0361] In certain embodiments, the methods and compositions disclosed herein are used to treat endometrial carcinoma. As an example, the methods and compositions disclosed herein can be used in combination with lenvatinib, for the treatment of subjects with advanced endometrial carcinoma that is not MSI-H or dMMR, who have disease progression following prior systemic therapy and are not candidates for curative surgery or radiation.

[0362] In certain embodiments, the methods and compositions disclosed herein are used to treat Tumor Mutational Burden-High (TMB-H) Cancer. As an example, the methods and compositions disclosed herein can be used to treat TMB-H cancer in subjects with unresectable or metastatic tumor mutational burden-high (e.g., ≥10 mutations / megabase (mut / Mb)) solid tumors, as determined by an FDA-approved 94 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 test, that have progressed following prior treatment and who have no satisfactory alternative treatment options.

[0363] In certain embodiments, the methods and compositions disclosed herein are used to treat Cutaneous Squamous Cell Carcinoma (cSCC). As an example, the methods and compositions disclosed herein can be used to treat cSCC in subjects with recurrent or metastatic cutaneous squamous cell carcinoma that is not curable by surgery or radiation.

[0364] In certain embodiments, the methods and compositions disclosed herein are used to treat Triple- Negative Breast Cancer (TNBC). As an example, the methods and compositions disclosed herein can be used in combination with chemotherapy, for the treatment of subjects with locally recurrent unresectable or metastatic TNBC whose tumors express PD-L1 (e.g., Combined Positive Score (CPS) ≥10) as determined by an FDA approved test.

[0365] In certain embodiments, the methods and compositions disclosed herein can be used to treat Merkel cell carcinoma (MCC). As an example, a combination comprising Avelumab can be used to treat MCC in subjects with metastatic MCC.

[0366] In certain embodiments, the methods and compositions disclosed herein can be used to treat Urothelial Carcinoma (UC). As an example, a combination comprising avelumab can be used to treat UC in subjects with locally advanced or metastatic UC who have disease progression during or following platinum-containing chemotherapy. As another example, a combination comprising avelumab can be used to treat UC in subjects with locally advanced or metastatic UC who have disease progression within 12 months of neoadjuvant or adjuvant treatment with platinum-containing chemotherapy.

[0367] In certain embodiments, the methods and compositions disclosed herein can be used to treat Renal Cell Carcinoma (RCC). As an example, a combination comprising avelumab and axitinib can be used in a subject with advanced RCC.

[0368] In certain embodiments, the methods and compositions disclosed herein can be used to treat urothelial carcinoma (UC). As an example, a combination comprising Durvalumab can be used to treat UC in subjects with locally advanced or metastatic urothelial carcinoma who have disease progression during or following platinum-containing chemotherapy. As another example, a combination comprising Durvalumab can be used to treat UC in subjects with locally advanced or metastatic urothelial carcinoma who have disease progression within 12 months of neoadjuvant or adjuvant treatment with platinum- containing chemotherapy.

[0369] In certain embodiments, the methods and compositions disclosed herein can be used to treat non- small cell lung cancer (NSCLC). As an example, a combination comprising Durvalumab can be used to 95 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 treat NSCLC in subjects with unresectable, Stage III non-small cell lung cancer (NSCLC) whose disease has not progressed following concurrent platinum-based chemotherapy and radiation therapy.

[0370] In certain preferred embodiments, the methods and compositions disclosed herein can be used to treat small cell lung cancer (SCLC). As an example, a combination comprising Durvalumab can be used in combination with etoposide and either carboplatin or cisplatin, as first-line treatment of adult subjects with extensive-stage small cell lung cancer (ES-SCLC).

[0371] In certain preferred embodiments, the methods and compositions disclosed herein can be used to treat urothelial carcinoma (UC). As an example, a combination comprising Atezolizumab can be used to treat UC in adult subjects with locally advanced or metastatic urothelial carcinoma who are not eligible for cisplatin-containing chemotherapy and whose tumors express PD-L1 (e.g., PD-L1 stained tumor- infiltrating immune cells [IC] covering ≥ 5% of the tumor area), as determined by an FDA-approved test, or are not eligible for any platinum-containing chemotherapy regardless of PD-L1 status, or have disease progression during or following any platinum-containing chemotherapy, or within 12 months of neoadjuvant or adjuvant chemotherapy.

[0372] In certain embodiments, the methods and compositions disclosed herein can be used to treat NSCLC. As an example, a combination comprising Atezolizumab can be used to treat NSCLC in adult subjects with metastatic NSCLC whose tumors have high PD-L1 expression (e.g., PD-L1 stained ≥ 50% of tumor cells [TC ≥ 50%] or PD-L1 stained tumor-infiltrating immune cells [IC] covering ≥ 10% of the tumor area [IC ≥ 10%]), as determined by an FDA-approved test, with no EGFR or ALK genomic tumor aberrations. As another example, a combination comprising Atezolizumab can be used in combination with bevacizumab, paclitaxel, and carboplatin, for the first-line treatment of adult subjects with metastatic non-squamous NSCLC with no EGFR or ALK genomic tumor aberrations. As another example, a combination comprising Atezolizumab can be used in combination with paclitaxel protein-bound and carboplatin for the first-line treatment of adult subjects with metastatic non-squamous NSCLC with no EGFR or ALK genomic tumor aberrations. As another example, a combination comprising Atezolizumab can be used to treat NSCLC in adult subjects with metastatic NSCLC who have disease progression during or following platinum-containing chemotherapy.

[0373] In certain embodiments, the methods and compositions disclosed herein can be used to treat triple negative breast cancer (TNBC). As an example, a combination comprising Atezolizumab can be used in combination with paclitaxel protein-bound for the treatment of adult subjects with unresectable locally advanced or metastatic TNBC whose tumors express PD-L1 (e.g., PD-L1 stained tumor-infiltrating immune cells [IC] of any intensity covering ≥ 1% of the tumor area), as determined by an FDA approved test. 96 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0374] In certain embodiments, the methods and compositions disclosed herein can be used to treat small cell lung cancer (SCLC). As an example, a combination comprising Atezolizumab can be used in combination with carboplatin and etoposide, for the first-line treatment of adult subjects with extensive- stage small cell lung cancer (ES-SCLC).

[0375] In certain embodiments, the methods and compositions disclosed herein can be used to treat endometrial cancer. As an example, a combination comprising Dostarlimab can be used to treat endometrial cancer in adult subjects with mismatch repair deficient (dMMR) recurrent or advanced endometrial cancer, as determined by an FDA-approved test, that has progressed on or following prior treatment with a platinum-containing regimen.

[0376] In certain embodiments, the methods and compositions disclosed herein can be used to treat cutaneous squamous cell carcinoma (CSCC). As an example, a combination comprising Cemiplimab-rwlc can be used to treat CSCC in subjects with metastatic cutaneous squamous cell carcinoma (mCSCC) or locally advanced CSCC (laCSCC) who are not candidates for curative surgery or curative radiation.

[0377] In certain embodiments, the methods and compositions disclosed herein can be used to treat basal cell carcinoma (BCC). As an example, a combination comprising Cemiplimab-rwlc can be used to treat BCC in subjects with locally advanced BCC (laBCC) previously treated with a hedgehog pathway inhibitor or for whom a hedgehog pathway inhibitor is not appropriate.

[0378] In certain embodiments, the methods and compositions disclosed herein can be used to treat NSCLC. As an example, a combination comprising Cemiplimab-rwlc can be used to treat NSCLC in subjects whose tumors have high PD-L1 expression (e.g., Tumor Proportion Score (TPS) ≥ 50%) as determined by an FDA-approved test, with no EGFR, ALK or ROS1 aberrations, and is locally advanced where subjects are not candidates for surgical resection or definitive chemoradiation, or metastatic.

[0379] In certain embodiments, the methods and compositions disclosed herein can be used to treat melanoma. As an example, a combination comprising Nivolumab can be used to treat melanoma in subjects with unresectable or metastatic melanoma, as a single agent or in combination with ipilimumab. As another example, a combination comprising Nivolumab can be used to treat melanoma in subjects with melanoma with lymph node involvement or metastatic disease who have undergone complete resection, in the adjuvant setting.

[0380] In certain embodiments, the methods and compositions disclosed herein can be used to treat NSCLC. As an example, a combination comprising Nivolumab can be used to treat NSCLC in adult subjects with metastatic non-small cell lung cancer expressing PD-L1 (≥1%) as determined by an FDA- approved test, with no EGFR or ALK genomic tumor aberrations, as first-line treatment in combination with ipilimumab. As another example, a combination comprising NSCLC can be used to treat melanoma 97 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 in adult subjects with metastatic or recurrent non-small cell lung cancer with no EGFR or ALK genomic tumor aberrations as first-line treatment, in combination with ipilimumab and 2 cycles of platinum- doublet chemotherapy. As another example, a combination comprising NSCLC can be used to treat melanoma in subjects with metastatic non-small cell lung cancer and progression on or after platinum- based chemotherapy.

[0381] In certain embodiments, the methods and compositions disclosed herein can be used to treat malignant pleural mesothelioma. As an example, a combination comprising Nivolumab can be used to treat malignant pleural mesothelioma in adult subjects with unresectable malignant pleural mesothelioma, as first-line treatment in combination with ipilimumab.

[0382] In certain embodiments, the methods and compositions disclosed herein can be used to treat RCC. As an example, a combination comprising Nivolumab can be used to treat RCC in subjects with intermediate or poor risk advanced renal cell carcinoma, as a first-line treatment in combination with ipilimumab. As another example, a combination comprising Nivolumab can be used to treat RCC in subjects with advanced renal cell carcinoma, as a first-line treatment in combination with cabozantinib. As another example, a combination comprising Nivolumab can be used to treat RCC in subjects with advanced renal cell carcinoma who have received prior anti-angiogenic therapy.

[0383] In certain embodiments, the methods and compositions disclosed herein can be used to treat classical Hodgkin lymphoma (cHL). As an example, a combination comprising Nivolumab can be used to treat cHL in adult subjects with cHL that has relapsed or progressed after autologous hematopoietic stem cell transplantation (HSCT) and brentuximab vedotin, or 3 or more lines of systemic therapy that includes autologous HSCT.

[0384] In certain embodiments, the methods and compositions disclosed herein can be used to treat squamous cell carcinoma of the head and neck (SCCHN). As an example, a combination comprising Nivolumab can be used to treat SCCHN in subjects with recurrent or metastatic squamous cell carcinoma of the head and neck with disease progression on or after a platinum-based therapy.

[0385] In certain embodiments, the methods and compositions disclosed herein can be used to treat urothelial carcinoma (UC). As an example, a combination comprising Nivolumab can be used to treat UC in subjects with locally advanced or metastatic urothelial carcinoma who have disease progression during or following platinum-containing chemotherapy or have disease progression within 12 months of neoadjuvant or adjuvant treatment with platinum-containing chemotherapy.

[0386] In certain embodiments, the methods and compositions disclosed herein can be used to treat colorectal cancer. As an example, a combination comprising Nivolumab can be used to treat colorectal cancer in subjects with microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR) 98 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 metastatic colorectal cancer that has progressed following treatment with a fluoropyrimidine, oxaliplatin, and irinotecan, as a single agent or in combination with ipilimumab.

[0387] In certain embodiments, the methods and compositions disclosed herein can be used to treat hepatocellular carcinoma (HCC). As an example, a combination comprising Nivolumab can be used to treat HCC in subjects with HCC who have been previously treated with sorafenib, as a single agent or in combination with ipilimumab.

[0388] In certain embodiments, the methods and compositions disclosed herein can be used to treat esophageal squamous cell carcinoma (ESCC). As an example, a combination comprising Nivolumab can be used to treat ESCC in subjects with unresectable advanced, recurrent or metastatic esophageal squamous cell carcinoma after prior fluoropyrimidine- and platinum-based chemotherapy.

[0389] In certain preferred embodiments, a combination comprising Camrelizumab can be used to treat cHL.

[0390] In certain preferred embodiments, a combination comprising Tislelizumab can be used to treat non-squamous non-small cell lung cancer. In certain preferred embodiments, a combination comprising Tislelizumab can be used to treat hepatocellular carcinoma (HCC).

[0391] In certain preferred embodiments, a combination comprising Toripalimab can be used to treat urothelial carcinoma. In certain preferred embodiments, a combination comprising Toripalimab can be used to treat melanoma. In certain preferred embodiments, a combination comprising Toripalimab can be used to treat nasopharyngeal carcinoma (NPC).

[0392] In certain preferred embodiments, a combination comprising Sintilimab can be used to treat non- squamous non-small cell lung cancer. In certain preferred embodiments, a combination comprising Sintilimab can be used to treat cHL.

[0393] The cancer to be treated using the methods and compositions of this disclosure can be metastatic cancer. The methods and compositions disclosed herein can be used to treat metastatic renal clear cell carcinoma or metastatic cutaneous malignant melanoma.

[0394] In some embodiments, provided herein is a method of enhancing an immune response in a subject in need thereof by administering an effective amount of an inducible IL-10 prodrug provided herein, or nucleic acid encoding the same, to the subject. The enhanced immune response may prevent, delay, or treat the onset of cancer, a tumor, or a viral disease. Without being bound by theory, the inducible IL-10 prodrug enhances the immune response by activating the innate and adaptive immunitiesThis also disclosure relates to a therapeutic combination of any of the inducible IL-10 prodrugs disclosed herein, or nucleic acids encoding the same, in combination with checkpoint inhibitors (e.g., PDL-1, PD-1). The 99 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 inducible IL-10 prodrugs and encoding nucleic acids disclosed herein can be combined with any desired anti-PD-1 antibody or any desired anti-PD-L1 antibody.

[0395] This disclosure further relates to methods for treating cancer using a combination therapy comprising an inducible IL-10 prodrug or nucleic acid encoding the same in combination with an anti- PD-1 antibody or an anti-PD-L1 antibody as disclosed herein, and to pharmaceutical compositions for use in such methods, including pharmaceutical compositions that contain an inducible IL-10 prodrug or nucleic acid encoding the same in combination an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0396] Exemplary anti-PD-1 antibodies that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same include, but are not limited to, AMP-224 (AstraZenica), 609A (3SBio), 704 (3SBio), 705 (3SBio), ABBV-181 (AbbVie), ADU-1503 / bion-004 (Chinook Therapeutics), AGEN2034 / balstilimab (Agenus), AK103 (Akeso), AK104 (Akeso), AK112 (Akeso), AK123 (Akeso), AMG 256 (Amgen), AMG 404 (Amgen), ANB030 (AnaptysBio), ANKEBIO Anti-PD1 product (Anhui Anke Biotechnology), Anti PD-1 / Anti-CD47 (DiNonA), ASKG915 (Ask Gene Pharmaceuticals), AV- MEL-1 (Aivita Biomedical), BCD-100 (Biocad CJSC), BI 754091 (Boehringer Ingelheim), BiCKI-IL-7 (OSE Immunotherapeutics), Boehringer-PD-1-unknown (Boehringer Ingelheim), BSK-050K01 (Biosion), Camrelizumab (Jiangsu Hengrui Medicine), CB201 (Crescendo Biologics), CB213 (Crescendo Biologics), CC-90006 (AnaptsBio), cetrelimab (J&J), chPD1 (Kiromic Biopharma), CMAB819 (Mabpharm), CS1003 (CStone Pharmaceuticals), CS17938 (Shenzhen Chipscreen Biosciences), CTX- 8371 (Compass Therapeutics), CX-072 (CytomX Therapeutics), CX-188 (CytomX Therapeutics), cypalizumab (Harbin Gloria Pharmaceuticals), DB004 (DotBio), EMB02 (EpimAb Biotherapeutics), Geptanblimab / genolimzumab (Apollomics), GS19 (Suzhou Zelgen Biopharmaceuticals), HLX10 (Shanghai Henlius Biotech), HX008 (Taizhou HanZhong Pharmaceuticals), HY003 (Juventas Cell Therapy), IBI315 / BH2950 (Innovent Biologics), IBI318 (Innovent Biologics), IBI319 (Innovent Biologics), IMM1802 (ImmuneOnco Biopharma), IMT200 (TrueBinding), Jemperli / dostarlimab (AnaptysBio), JTX-4014 (Jounce Therapeutics), Keytruda / pembrolizumab (Merck), LBL-006 (Nanjing Leads Biolabs), Libtayo / cemiplimab-rwlc (Regeneron Pharmaceuticals), LVGN3616 (Lyvgen Biopharma), LXF821 (Novartis), LY01015 (Luye Pharma Group), LY3462817 (Eli Lilly), MCLA-134 (Merus N.V.), MEDI5752 (AstraZenica), NIR178 (Novartis), ONCR-177 (Oncorus), ONO-4685 (Ono Pharmaceutical), Opdivo / nivolumab (Ono Pharmaceutical), MGD019 (MacroGenius), PD1-GDT CAR- T (Kiromic Biopharma), penpulimab (Akeso), PSB205 (Qilu Puget Sound Biotherapeutics), PT-001 (Merck), PT627 (Merck), RB-M1 (Refuge Biotechnologies), Retifanlimab (MacroGenics), RG6139 (Roche), RG6279 (Roche), RTX-002 (RubrYc Therapeutics), sasanlimab (Pfizer), Servier-PD1xLAG3- unknown (Servier), SL-279252 / TAK-252 (Shattuck Labs), Sofusa anti-PD1 (Sorrento Therapeutics), 100 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 spartalizumab (Novartis), SSI-361 (Lyvgen Biopharma), Sym021 (Servier), Tebotelimab (MacroGenics), tislelizumab (BeiGene), TSR-075 (AnaptsBio), Tuhura-DO / PD-1-unknown (Tuhura Biopharma), toripalimab (Shanghai Junshi Biosciences), sintilimab (Innovent Biologics), Unicar-CAR-T&PD-1- unknown (Shanghai Unicar-Therapy Bio-Medicine Technology), Xdivane (Xbrane Biopharma), XmAb20717 (Xencor), XmAb23104 (Xencor), YBL-006 (Y-Biologics), and zimberelimab (Arcus Biosciences).

[0397] The anti-PD-1 antibody that can be combined with the inducible cytokine prodrugs is typically an approved anti-PD-1 antibody. Approved anti-PD-1 antibodies include, but are not limited to, pembrolizumab (KEYTRUDA), dostarlimab (JEMPERLI), cemiplimab-rwlc (LIBATYO), nivolumab (OPDIVO), camrelizumab, tislelizumab, toripalimab, and sintilimab (TYVYT).

[0398] Exemplary anti-PD-L1 antibodies that can be combined with the inducible cytokine prodrugs include, but are not limited to, A167 (Sichuan Kelun), ABL501 (ABL Bio), ABL503 (ABL Bio), ABSK041 (Abbisko Therapeutics), ACE1708 (Acepodia), ACE-NK-PDL1 (Acepodia), ADG104 (Adagene), AK106 (Akeso), ALPN-202 (Alpine Immune Sciences), AN4005 (Adlai Nortye Biopharma), BMS-936559 / MDX-1105 (BMS), APL-502 / TQB2450 (Apollomics), Arbutus-PD-L1-unknown (Arbutus Biopharma), ASC22 (Ascletis Pharma), ATG-101 (Antengene), AVA-004 (Avacta Group), AVA021 (Avacta Group), AVA027 (Avacta Group), AVA-040-100 (Avacta Group), AVA04-Vbp (Avacta Group), Bavencio / avelumab (Merck), BCD-135 (Biocad CJSC), BGB-A333 (BeiGene), Bintrafusp alfa / GSK4045154 (Merck), CA-170 / aupm-170 (Dr. Reddy’s Laboratories), CCX559 (ChemoCentryx), CDR101 (CDR-Life), cosibelimab (Checkpoint Therapeutics), CTX-8371 (Compass Therapeutics), DiNonA-Solid Tumors-unknown (DiNonA), DR30207 (Zhejiang Doer Biologics), DuoBody-PD-L1x4-1BB (Ligand Pharmaceuticals), envafolimab (Alphamab Oncology), EPIM-001 (Elpis Biopharmaceuticals), ES101 (Elpiscience Biopharma), INBRX-105 (Inhibrx), FAZ053 (Novartis), FS118 (F-star Therapeutics), GB262 (Genor Biopharma), GS-4224 (Gilead), GT900008 (Kintor Pharmaceuticals), GX-P2 (Genexine), Hamni-PS-L1 / CD47-Unknown (Hanmi Pharmaceutical), HBM7015 (HBM Holdings), HBM9167 (HBM Holdings), HLX20 (Shanghai Henlius Biotech), HTI- 1088 (Jiangsu Hengrui Medicine), IBI318 (Innovent Biologics), IBI322 (Innovent Biologics), IBI323 (Innovent Biologics), IGM-7354 (IGM Biosciences), IMC-001 (Sorrento Therapeutics), Imfinzi / durvalumab (AstraZenica), IMM25 (ImmuneOnco Biopharma), IMM2502 (ImmuneOnco Biopharma), IMM2503 (ImmuneOnco Biopharma), IMM2504 (ImmuneOnco Biopharma), INCB86550 (Incyte), IO103 (IO Biotech), JS003 (Shanghai Junshi Biosciences), Jubilant-PD-L1-unknown (Jubilant Therapeutics), KD033 (Kadmon Holdings), KN046 (Alphamab Oncology), KY1003 (Sanofi), KY1043 (Sanofi), LY3300054 (Eli Lilly), LY3415244 (Eli Lilly), MRNA-6981 (Moderna), MSB2311 (Transcenta 101 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 Holding), MT-6035 (Molecular Templates), ND021 / NM21-1480 (Numab Therapeutics), OX001R (Oxford BioTherapeutics), PD-L1 based BsAbs (I-Mab), PD-L1 Boltbody ISAC (Bolt Biotherapeutics), PDL-GEX (Glycotope GmbH), PMC-122 (PharmAbcine), PMI06 (D&D Pharmatech), Protheragen-RV- scFv-PDL1-unknown (Protheragen), PRS-344 (Pieris Pharmaceuticals), Q-1802 (Merck), RC98 (Yantai Rongchang Pharmaceutical), RV-scFv-PDL1 (Protheragen), SenI_TAAx22P (Hebei Senlang Biotechnology), SHC020 (Nanjing Sanhome Pharmaceutical), sugemalimab (Ligand Pharmaceuticals), atezolizumab (Roche), TST005 (Transcenta Holding), TT-01 (Topmunnity Therapeutics), TTX-siPDL1 (TransCode Therapeutics), UniCAR-T-PD-L1 (GEMoaB monoclonals), Vaximm (VXM10), and YBL- 013 (Y-Biologics).

[0399] The anti-PD-L1 antibody that can be combined with the inducible IL-10 prodrugs or nucleic acid encoding the same, include is typically an approved anti-PD-L1 antibody. Approved anti-PD-1 antibodies include, but are not limited to, avelumab (BAVENCIO), durvalumab (IMFINZI), and atezolizumab (TECENTRIQ).

[0400] The disclosure relates to methods for treating cancer comprising administering to a subject in need thereof an effective amount of a combination therapy that includes an inducible IL-10 prodrug or nucleic acid encoding the same and an anti-PD-1 antibody or an anti-PD-L1 antibody. The inducible IL- 10 prodrug nucleic acid encoding the same and an anti-PD-1 antibody or an anti-PD-L1 antibody are administered to the subject so that there is overlap of the pharmacological activities of the two therapeutic agents. Accordingly, the inducible cytokine prodrug can be administered before, after, concurrently, or periprocedurally with an anti-PD-1 antibody or an anti-PD-L1 antibody. In some practices of the methods, an anti-PD-1 antibody or an anti-PD-L1 antibody is administered before the inducible IL- prodrug or nucleic acid encoding the same. In some practices of the methods, the anti-PD-1 antibody or the anti-PD- L1 antibody is administered, then after administration is completed, the inducible IL- prodrug nucleic acid encoding the same is administered.

[0401] The inducible IL-10 prodrug nucleic acid encoding the same and the anti-PD-1 / anti-PD- L1antibody are typically administered systemically, for example by intravenous injection or preferably intravenous infusion. Other types of administration can be used, such as orally, parenterally, intravenous, intravenously, intra-articularly, intraperitoneally, intramuscularly, subcutaneously, intracavity, transdermally, intrahepatically, intracranially, nebulization / inhalation, by installation via bronchoscopy, or intratumorally.

[0402] If desired, additional therapeutic agents can be administered to the subject. Typically such additional therapeutic agents are anti-cancer agents such as chemotherapeutic agents (e.g., adriamycin, cerubidine, bleomycin, alkeran, velban, oncovin, fluorouracil, thiotepa, methotrexate, bisantrene, 102 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 noantrone, thiguanine, cytaribine, procarabizine), other immunocheck point inhibitors (e.g., anti-CTLA4 (ipilimumab (YERVOY)), other cytokines (such as IL-12, inducible IL-12 prodrugs, IFN, inducible IFN prodrugs, IL-2, inducible IL-2 prodrugs, IL-21, inducible IL-21 prodrugs), angiogenesis inhibitors, antibody-drug conjugates (e.g., trastuzumab emtansine (KADCYLA), trastuzumab deruxtecan (ENHERTU), enfortumab vedotin (PADCEV), sacituzumab govitecan (TRODELVY), cellular therapies (e.g., CAR-T, TCT-T, T-cell therapy, such as tumor infiltrating lymphocyte (TIL) therapy), oncolytic viruses, radiation therapy and / or small molecules.

[0403] An additional therapeutic agent can be, for example, pemetrexed, a platinum chemotherapeutic agent, carboplatin, paclitaxel, protein-bound paclitaxel, fluorouracil, a fluoropyrimidine-based chemotherapeutic agent, or axitinib. H. Definitions

[0404] All publications and patents cited in this disclosure are incorporated by reference in their entirety. To the extent the material incorporated by reference contradicts or is inconsistent with this specification, the specification will supersede any such material. The citation of any references herein is not an admission that such references are prior art to the present disclosure. When a range of values is expressed, it includes embodiments using any particular value within the range. Further, reference to values stated in ranges includes each and every value within that range. All ranges are inclusive of their endpoints and combinable. When values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise. The use of “or” will mean “and / or” unless the specific context of its use dictates otherwise.

[0405] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein. The techniques and procedures described or referenced herein are generally well understood and commonly employed using conventional methodologies by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 4th ed. (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer-defined protocols and conditions unless otherwise noted. 103 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320

[0406] As used herein, the singular forms “a,” “an,” and “the” include plural forms unless the context clearly indicates otherwise. The terms “include,” “such as,” and the like are intended to convey inclusion without limitation, unless otherwise specifically indicated.

[0407] Unless otherwise indicated, the terms "at least," "less than," and "about," or similar terms preceding a series of elements, or a range are to be understood to refer to every element in the series or range. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

[0408] As used herein, the terms “activatable,” “activate,” “induce,” and “inducible” refers to a polypeptide complex that has an attenuated activity form (e.g., attenuated receptor binding and / or agonist activity) and an activated form. The polypeptide complex is activated by protease cleavage of the linker that causes the blocking element and half-life extension element to dissociate from the polypeptide complex. The induced / activated polypeptide complex can bind with increased affinity / avidity to the IL-10 receptor.

[0409] The terms “antibody” and “immunoglobulin” are used interchangeably herein. An antibody or immunoglobulin, as used herein, is intended to refer to immunoglobulin molecules comprised of two heavy (H) chains. Typically, antibodies in mammals (e.g., humans, rodents, and monkey’s) comprise four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CHI, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. Antibodies can include, for example, monoclonal antibodies, recombinantly produced antibodies, monospecifi...

Claims

Attorney Docket No.: 761146.192320 CLAIMS 1. An inducible IL-10 prodrug comprising at least one of each of: a) an IL-10 polypeptide [A]; b) an IL-10 blocking element [D]; c) a protease-cleavable polypeptide linker [L]; wherein the IL-10 polypeptide and the IL-10 blocking element are operably linked by the protease- cleavable polypeptide linker and the inducible IL-10 has attenuated IL-10 receptor activating activity, wherein the IL-10-receptor activating activity of the inducible IL-10 prodrug is at least about 10X less than the IL-10 receptor activating activity of the polypeptide that contains the IL-10 polypeptide that is produced by cleavage of the protease cleavable linker.

2. The inducible IL-10 prodrug of claim 1, wherein [A] comprises one IL-10 polypeptide, two IL-10 polypeptides, or three IL-10 polypeptides.

3. The inducible IL-10 prodrug of claim 2, wherein the inducible IL-10 prodrug comprises one IL- 10 polypeptide.

4. The inducible IL-10 prodrug of claim 2, wherein the inducible IL-10 prodrug comprises two IL- 10 polypeptides.

5. The inducible IL-10 prodrug of claim 2, wherein the inducible IL-10 prodrug comprises three IL- 10 polypeptides.

6. The inducible IL-10 prodrug of any one of claims 4 or 5, wherein the two IL-10 polypeptides or three IL-10 polypeptides are operably linked by an optionally protease-cleavable linker.

7. The inducible IL-10 prodrug of claim 2, wherein [A] has the formula: [A1]-[L3]-[A1], wherein [A1] is an IL-10 polypeptide; and [L3] is a polypeptide linker that is optionally protease cleavable. 178 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 8. The inducible IL-10 prodrug of any one of the preceding claims, wherein the blocking element [D] is an IL-10 blocking element that also extends in vivo half-life.

9. The inducible IL-10 prodrug of any one the preceding claims, wherein the IL-10 blocking element is human serum albumin, or an antigen binding protein or polypeptide which binds human serum albumin or a fragment thereof.

10. The inducible IL-10 prodrug of any one of the preceding claims, wherein the IL-10 blocking element sterically inhibits or blocks activation and / or binding of the IL-10 polypeptide to its cognate receptor.

11. The inducible IL-10 prodrug of any one of the preceding claims, wherein the inducible IL-10 prodrug comprises two blocking elements.

12. The inducible IL-10 prodrug of any one of the preceding claims, wherein the inducible IL-10 prodrug comprises three blocking elements.

13. The inducible IL-10 prodrug of any one of the preceding claims, wherein the IL-10 blocking element binds the human IL-10 polypeptide.

14. The inducible IL-10 prodrug of any one of the preceding claims, wherein the IL-10 blocking element comprises a ligand binding domain or fragment of a cognate receptor for the IL-10, an antibody or antibody fragment that binds the IL-10.

15. The inducible IL-10 prodrug of claim 14, wherein the antibody or antigen-binding fragment is a single domain antibody, a Fab, or a scFv that binds the IL-10 polypeptide.

16. The inducible IL-10 prodrug of claim 14, wherein the cognate receptor for IL-10 is the IL10Ralpha or IL10Rbeta.

17. The inducible IL-10 prodrug of any one of the preceding claims, further comprising an optional half-life extension element [H]. 179 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 18. The inducible IL-10 prodrug of claim 17, wherein the half-life extension element comprises a serum albumin binding domain, a serum albumin, transferrin, or an immunoglobulin Fc, or fragment thereof.

19. The inducible IL-10 prodrug of any one of the preceding claims, wherein the optionally protease- cleavable linker is not protease cleavable.

20. The inducible IL-10 prodrug of any one of the preceding claims, wherein each protease-cleavable polypeptide linker independently comprises a sequence that is capable of being cleaved by a protease selected from the group consisting of a kallikrein, a thrombin, a chymase, a carboxypeptidase, a cathepsin, an elastase, a PR-3, a granzyme, a calpain, a matrix metalloproteinase (MMP), a fibroblast activation protein (FAP), an ADAM metalloproteinase, a plasminogen activator, a caspase, a tryptase, and a tumor cell surface protease.

21. The inducible IL-10 prodrug of claim 20, wherein each protease-cleavable polypeptide independently comprises two or more cleavage sites for the same protease, or two or more cleavage sites that are cleaved by different proteases or at least one of the protease-cleavable polypeptides comprises a cleavage site for two or more different proteases.

22. The inducible IL-10 prodrug of claim 20 or 21, wherein the cathepsin is cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, cathepsin S, or cathepsin G.

23. The inducible IL-10 prodrug of claim 20 or 21, wherein the matrix metalloprotease (MMP) is MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, MMP19, or MMP20.

24. The inducible IL-10 prodrug of any one of the preceding claims, wherein the activating activity is assessed using a HEK-Blue reporter cell assay.

25. The inducible IL-10 prodrug of any one of the preceding claims, wherein IL-10 is free to dissociate from the IL-10 blocking element and when present, the half-life extension element after the protease-cleavable sequence is cleaved by a protease. 180 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 26. The inducible IL-10 prodrug of any one of the preceding claims, wherein the inducible IL-10 prodrug has the formula: [D1]-[L1]-[A]-[L1]-[D2]; or [D1]-[L1]-[A]-[L2]-[A]-[L1]-[ D2]; or ; wherein, , [D2] and [D3] are each, independently, a blockinga polypeptide linker, wherein L1 is a protease-cleavable polypeptide linker, and L2 is a polypeptide linker that is optionally protease cleavable.

27. The inducible IL-10 prodrug of claim 1, further comprising a half-life extension element [H] and having the formula: [A]-[L1]-[D]-[L2]-[H] or [A]-[L1]-[H]-[L2]-[D] or [H]-[L1]-[A]-[L4]-[D] or [D]-[L1]-[A]-[L4]-[H] or [H]-[L2]-[D]-[L1]-[A] or [D]-[L2]-[H]-[L1]-[A] wherein [L1] and [L2] and [L4] are each independently a polypeptide linker, wherein [L1] is a protease-cleavable polypeptide linker, [L2] is polypeptide linker that is optionally protease cleavable, and [L4] is a protease- cleavable polypeptide linker.

28. The inducible IL-10 prodrug of claim 26 or 27, wherein [A] has the formula: [A1]-[L3]-[A2], wherein [A1] and [A2] are each independently an IL-10 polypeptide, wherein [A1] and [A2] can have the same or different amino acid sequence; and [L3] is a polypeptide linker that is optionally protease cleavable.

29. The inducible IL-10 prodrug of claim 26 or 27, wherein [L2] is a protease cleavable linker.

30. The inducible polypeptide of any one of claim 26 or 27, wherein [L1] is a substrate for a first protease and [L2] is a substrate for a second protease. 181 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 31. The inducible IL-10 prodrug of any one of the preceding claims, wherein the protease-cleavable polypeptide linker comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249 or 259, or an amino acid sequence that has at least 90% identity to SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 195-201, 249 or 259.

32. The inducible IL-10 prodrug of any one of the preceding claims, wherein the protease-cleavable polypeptide linker comprises SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 195-201, 249 or 259 or a functional variant of SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249 or 259.

33. The inducible IL-10 prodrug of any one of the proceeding claims, wherein the IL-10 polypeptide is an IL-10 dimer.

34. The inducible IL-10 prodrug of any one of the preceding claims, wherein the IL-10 polypeptide comprises two IL-10 polypeptides, wherein the two IL-10 polypeptides form a dimer by covalent or intramolecular binding or optionally operably linked by a polypeptide linker.

35. The inducible IL-10 prodrug of any one of the preceding claims, wherein the protease-cleavable polypeptide linker comprises SEQ ID NO: 37, SEQ ID NO: 31, SEQ ID NO: 195, SEQ ID NO: 249, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, or SEQ ID NO:

115.

36. The inducible IL-10 prodrug of any one of the preceding claims, wherein the protease-cleavable polypeptide linker comprises SEQ ID NO:

195.

37. A nucleic acid encoding the inducible IL-10 prodrug of any of claims 1-36.

38. The nucleic acid of claim 37, wherein the nucleic acid comprises a circular vector.

39. The nucleic acid of claim 37, wherein the nucleic acid comprises DNA.

40. The nucleic acid of claim 37, wherein the nucleic acid comprises RNA. 182 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 41. An expression vector comprising the nucleic acid of any one of claims 37-40.

42. An isolated host cell comprising the vector of claim 41.

43. A method of making a pharmaceutical composition, comprising culturing the host cell of claim 42 under suitable conditions for expression and collection of desired polypeptides.

44. A pharmaceutical composition comprising an effective amount of the inducible IL-10 prodrug of any of claims 1-36 or a nucleic acid of any one of claims 37-40.

45. A method for treating an inflammatory condition, comprising administering to a subject in need thereof an effective amount of the inducible IL-10 prodrug of any one of claims 1-36, a nucleic acid of any one of claims 37-40, the expression vector of claim 41, or the pharmaceutical composition of claim 44.

46. An inducible IL-10 prodrug comprising the amino acid sequence selected from the group consisting of SEQ ID NOs.: 59, 60, 62, 90, or 91 or an amino acid sequence that has at least about 80% identity to SEQ ID NOs.: 59, 60, 62, 90, or 91.

47. A nucleic acid encoding the inducible IL-10 prodrug of claim 46.

48. The nucleic acid of claim 47, wherein the nucleic acid comprises a circular vector.

49. The nucleic acid of claim 47, wherein the nucleic acid comprises DNA.

50. The nucleic acid of claim 47, wherein the nucleic acid comprises RNA.

51. An expression vector comprising the nucleic acid of any one of claims 47-50.

52. An isolated host cell comprising the vector of claim 51.

53. A method of making a pharmaceutical composition, comprising culturing the isolated host cell of claim 52 under suitable conditions for expression of the inducible IL-10 prodrug. 183 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 54. The method of claim 53, further comprising isolating the inducible IL-10 prodrug.

55. A pharmaceutical composition comprising the inducible IL-10 prodrug of claim 46 or nucleic acid of any one of claims 47-50.

56. A method for treating an inflammatory condition, comprising administering to a subject in need thereof an effective amount of the inducible IL-10 prodrug of claim 46, a nucleic acid of any one of claims 47-50, the expression vector of claim 51, or the pharmaceutical composition of claim 55.

57. A nucleic acid composition comprising one or more nucleic acid sequences encoding an inducible IL-10 prodrug comprising at least one of each of: a) an IL-10 polypeptide [A]; b) an IL-10 blocking element [D]; c) a protease-cleavable polypeptide linker [L]; wherein the IL-10 polypeptide and the IL-10 blocking element are operably linked by the protease- cleavable polypeptide linker and the inducible IL-10 has attenuated IL-10 receptor activating activity, wherein the IL-10-receptor activating activity of the inducible IL-10 prodrug is at least about 10X less than the IL-10 receptor activating activity of the polypeptide that contains the IL-10 polypeptide that is produced by cleavage of the protease cleavable linker.

58. The nucleic acid composition of claim 57, wherein the nucleic acid comprises a circular vector.

59. The nucleic acid composition of claim 57, wherein the nucleic acid comprises DNA.

60. The nucleic acid composition of claim 57, wherein the nucleic acid comprises RNA.

61. An expression vector comprising the nucleic acid composition of any one of claims 58-60.

62. An isolated host cell comprising the vector of claim 61.

63. An inducible IL-10 prodrug comprising an IL-10 polypeptide, a half-life extension element, an IL-10 blocking element, and a protease cleavable linker, wherein the IL-10 blocking element is an antigen 184 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 binding fragment of an antibody, wherein the antigen binding fragment comprises as separate components, at least an antigen-binding portion of an antibody light chain and at least an antigen-binding portion of a complementary antibody heavy chain, and the inducible IL-10 prodrug comprises: i. a first polypeptide comprising the IL-10 polypeptide, at least an antigen binding portion of an antibody light chain or an antigen binding portion of an antibody heavy chain, and the half-life- extension element; wherein the IL-10 polypeptide and the antigen binding portion of the antibody light chain or the antigen binding portion of the antibody heavy chain and / or half-life extension element are operably linked by the protease cleavable linker; and ii. a second polypeptide comprising at least an antigen binding portion of an antibody heavy chain that is complementary to the light chain in the first polypeptide, or an antibody light chain that is complementary to the heavy chain in the first polypeptide and together with said light chain forms an IL-10 binding site.

64. The inducible IL-10 prodrug of claim 63, wherein the first polypeptide has a formula of: [A]-[L1]-[D]-[L2]-[H] or [A]-[L1]-[H]-[L2]-[D] or [H]-[L1]-[A]-[L4]-[D] or [D]-[L1]-[A]-[L4]-[H] or [H]-[L2]-[D]-[L1]-[A] or [D]-[L2]-[H]-[L1]-[A] wherein, [A] is an interleukin 10 (IL-10) polypeptide; [H] is a half-life extension element; [L1] and [L2] and [L4] are each independently a polypeptide linker, wherein [L1] is a protease-cleavable polypeptide linker,[L2] is a polypeptide linker that is optionally protease cleavable, and [L4] is a protease-cleavable polypeptide linker; [D] is an IL-10 antigen binding fragment of an antibody; wherein the antigen binding fragment of the antibody is an antigen-binding portion of an antibody light chain or at least an antigen- binding portion of a complementary antibody heavy chain, and the inducible IL-10 prodrug is an attenuated IL-10 receptor agonist, but upon (i) cleavage of the L1 protease-cleavable polypeptide linker, or (ii) cleavage of both L1 and L2 when L2 is a protease cleavable polypeptide linker, the IL-10 polypeptide-containing fragment of the inducible IL-10 prodrug 185 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 that is produced is an IL-10 receptor agonist of comparable potency and half-life to the naturally occurring IL-10 polypeptide.

65. The inducible IL-10 prodrug of claim 63, wherein the second polypeptide comprises an antigen- binding portion of an antibody light chain or at least an antigen-binding portion of a complementary antibody heavy chain; wherein when the antigen-binding portion of an antibody light chain is on the first polypeptide chain, the antigen-binding portion of a complementary antibody heavy chain is on the second polypeptide chain, and wherein when the antigen-binding portion of a complementary antibody heavy chain is on the first polypeptide chain, the antigen-binding portion of an antibody light chain is on the second polypeptide.

66. The inducible IL-10 prodrug any one of claims 63-65, wherein [A] has the formula: [A1]-[L3]-[A2], wherein [A1] and [A2] are each independently an IL-10 polypeptide, wherein [A1] and [A2] can have the same or different amino acid sequence; and [L3] is a polypeptide linker that is optionally protease cleavable.

67. The inducible IL-10 prodrug of claim 66, wherein [L3] is not protease cleavable.

68. The inducible IL-10 prodrug of any one of claims 63-67, wherein each protease-cleavable polypeptide linker independently comprises a sequence that is capable of being cleaved by a protease selected from the group consisting of a kallikrein, a thrombin, a chymase, a carboxypeptidase, a cathepsin, an elastase, PR-3, a granzyme, a calpain, a matrix metalloproteinase (MMP), a fibroblast activation protein (FAP), an ADAM metalloproteinase, a plasminogen activator, a caspase, a tryptase, and a tumor cell surface protease.

69. The inducible IL-10 prodrug any one of claims 63-67, each protease-cleavable polypeptide independently comprises two or more cleavage sites for the same protease, or two or more cleavage sites that are cleaved by different proteases or at least one of the protease-cleavable polypeptides comprises a cleavage site for two or more different proteases. 186 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 70. The inducible IL-10 prodrug of claim 68, wherein the protease is selected from cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, cathepsin S, or cathepsin G.

71. The inducible IL-10 prodrug of claim 68, wherein the protease is selected from matrix metalloprotease (MMP) is MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, MMP19, or MMP20.

72. The inducible IL-10 prodrug of any one of claims 63-71, wherein the blocking element binds the IL-10 polypeptide.

73. The inducible IL-10 prodrug of any one of claims 63-72, wherein the half-life extension element is human serum albumin, an immunoglobulin Fc or fragment thereof, or an antigen binding polypeptide which binds human serum albumin.

74. The inducible IL-10 prodrug of claim 73, wherein the half-life extension element is an immunoglobulin Fc.

75. A nucleic acid encoding the inducible prodrug of any one of claims 63-74.

76. The nucleic acid claim 75, wherein the nucleic acid comprises a circular vector.

77. The nucleic acid of claim 75, wherein the nucleic acid comprises DNA.

78. The nucleic acid of claim 75, wherein the nucleic acid comprises RNA.

79. An expression vector comprising the nucleic acid of any one of claims 75-78.

80. An isolated host cell comprising the vector of claim 79.

81. A method of making a pharmaceutical composition, comprising culturing the host cell of claim 80 under suitable conditions for expression and collection of the inducible IL-10 prodrugs. 187 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 82. A pharmaceutical composition comprising an effective amount of the inducible IL-10 prodrugs of any of claims 63-74 or a nucleic acid of any one of claims 75-78.

83. A method for treating an inflammatory condition, comprising administering to a subject in need thereof an effective amount of the inducible IL-10 prodrug of any one of claims 63-74, or a nucleic acid of any one of claims 75-78, or the pharmaceutical composition of claim 82.

84. The inducible IL-10 prodrug of any one of claims 63-74, wherein the protease-cleavable peptide linker comprises an amino acid sequence selected from the group consisting of SEQ ID NOs.: 5, 9-11, 14- 35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249 or 259, or an amino acid sequence that has at least 90% identity to SEQ ID NOs.: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249 or 259.

85. The inducible IL-10 prodrug of any one of claims 63-74, wherein the protease-cleavable peptide linker comprises SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249 or 259 or a functional variant of SEQ ID NOs.: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195- 201, 249 or 259.

86. The inducible IL-10 prodrug of any one of claims 63-74, wherein the protease-cleavable peptide linker comprises SEQ ID NO:

198.

87. The inducible IL-10 prodrug of any one of claims 63-74, wherein the protease-cleavable peptide linker comprises SEQ ID NO:

195.

88. The inducible IL-10 prodrug of any one of claims 63-74, wherein the protease-cleavable peptide linker comprises SEQ ID NO:

37.

89. The inducible 1L-10 prodrug of any one of claims 63-74, wherein the protease-cleavable peptide linker comprises SEQ ID NO:

38.

90. The inducible IL-10 prodrug of any one of claims 63-74, wherein the protease-cleavable peptide linker comprises SEQ ID NO: 39, SEQ ID NO: 249, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, or SEQ ID NO:

115. 188 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 91. The inducible IL-10 prodrug of any one of claims 63-74, wherein the first polypeptide comprises the amino acid sequence of SEQ ID NOs.: 68, 72-76, 81-87, 92-104, 116-119, 123-126, or 622 and the second polypeptide can comprise the amino acid sequence of SEQ ID NOs.: 69-71.

92. A nucleic acid composition comprising one or more nucleic acid sequences encoding an inducible IL-10 prodrug comprising an IL-10 polypeptide, a half-life extension element, an IL-10 blocking element, and a protease cleavable linker, wherein the IL-10 blocking element is an antigen binding fragment of an antibody, wherein the antigen binding fragment comprises as separate components, at least an antigen- binding portion of an antibody light chain and at least an antigen-binding portion of a complementary antibody heavy chain, and the inducible IL-10 prodrug comprises: i. a first polypeptide comprising the IL-10 polypeptide, at least an antigen binding portion of an antibody light chain or an antigen binding portion of an antibody heavy chain, and the half-life- extension element; wherein the IL-10 polypeptide and the antigen binding portion of the antibody light chain or the antigen binding portion of the antibody heavy chain and / or half-life extension element are operably linked by the protease cleavable linker; and ii. a second polypeptide comprising at least an antigen binding portion of an antibody heavy chain that is complementary to the light chain in the first polypeptide, or an antibody light chain that is complementary to the heavy chain in the first polypeptide and together with said light chain forms an IL-10 binding site.

93. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element comprises Blocking Element 1 (SEQ ID NO: 127 / SEQ ID NO: 168), Blocking Element 2 (SEQ ID NO: 128 / SEQ ID NO: 169), Blocking Element 3 (SEQ ID NO: 129 / SEQ ID NO: 170), Blocking Element 4 (SEQ ID NO: 130 / SEQ ID NO: 171), Blocking Element 5 (SEQ ID NO: 131 / SEQ ID NO: 172), Blocking Element 6 (SEQ ID NO: 132 / SEQ ID NO: 173), Blocking Element 7 (SEQ ID NO: 133 / SEQ ID NO: 174), Blocking Element 8 (SEQ ID NO: 134 / SEQ ID NO: 175), Blocking Element 9 (SEQ ID NO: 135 / SEQ ID NO: 176), Blocking Element 10 (SEQ ID NO: 136 / SEQ ID NO: 177), Blocking Element 10 (SEQ ID NO: 136 / SEQ ID NO: 177), Blocking Element 11 (SEQ ID NO: 137 / SEQ ID NO: 178), Blocking Element 12 (SEQ ID NO: 138 / SEQ ID NO: 179), Blocking Element 13 (SEQ ID NO: 139 / SEQ ID NO: 180), Blocking Element 14 (SEQ ID NO: 140 / SEQ ID NO: 181), Blocking Element 15 (SEQ ID NO: 141 / SEQ ID NO: 182), Blocking Element 16 (SEQ ID NO: 142 / SEQ ID NO: 183), Blocking Element 17 (SEQ ID NO: 143 / SEQ ID NO: 184), Blocking Element 18 (SEQ ID NO: 189 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 144 / SEQ ID NO: 185), Blocking Element 19 (SEQ ID NO: 145 / SEQ ID NO: 186), Blocking Element 20 (SEQ ID NO: 146 / SEQ ID NO: 187), Blocking Element 21 (SEQ ID NO: 147 / SEQ ID NO: 188), Blocking Element 22 (SEQ ID NO: 148 / SEQ ID NO: 189), Blocking Element 23 (SEQ ID NO: 149 / SEQ ID NO: 190), Blocking Element 24 (SEQ ID NO: 150 / SEQ ID NO: 191), Blocking Element 25 (SEQ ID NO: 151 / SEQ ID NO: 192), Blocking Element 26 (SEQ ID NO: 152 / SEQ ID NO: 193), Blocking Element 27 (SEQ ID NO: 153 / SEQ ID NO: 194), Blocking Element 28 (SEQ ID NO: 154 / SEQ ID NO: 500), Blocking Element 29 (SEQ ID NO: 155 / SEQ ID NO: 501), Blocking Element 30 (SEQ ID NO: 156 / SEQ ID NO: 502), Blocking Element 31 (SEQ ID NO: 157 / SEQ ID NO: 503), Blocking Element 32 (SEQ ID NO: 158 / SEQ ID NO: 504), Blocking Element 33 (SEQ ID NO: 159 / SEQ ID NO: 505), Blocking Element 34 (SEQ ID NO: 160 / SEQ ID NO: 506), Blocking Element 35 (SEQ ID NO: 161 / SEQ ID NO: 507), Blocking Element 36 (SEQ ID NO: 162 / SEQ ID NO: 507), Blocking Element 37 (SEQ ID NO: 163 / SEQ ID NO: 508), Blocking Element 38 (SEQ ID NO: 164 / SEQ ID NO: 509), Blocking Element 39 (SEQ ID NO: 165 / SEQ ID NO: 510), Blocking Element 40 (SEQ ID NO: 166 / SEQ ID NO: 512), Blocking Element 41 (SEQ ID NO: 167 / SEQ ID NO: 513).

94. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 127 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 127, and a light chain constant region that comprises or consists of SEQ ID NO: 168 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

168.

95. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 128 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 128, and a light chain constant region that comprises or consists of SEQ ID NO: 169 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

169.

96. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 130 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 130, and a light chain constant region that comprises or consists of SEQ ID NO: 171 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

171. 190 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 97. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 139 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 139, and a light chain constant region that comprises or consists of SEQ ID NO: 180 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

180.

98. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 140 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 140, and a light chain constant region that comprises or consists of SEQ ID NO: 181 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

181.

99. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 144 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 144, and a light chain constant region that comprises or consists of SEQ ID NO: 185 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

185.

100. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 149 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 149, and a light chain constant region that comprises or consists of SEQ ID NO: 190 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

190.

101. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 149 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 149, and a light chain constant region that comprises or consists of SEQ ID NO: 190 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

190.

102. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 151 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 151, and a light chain constant 191 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 region that comprises or consists of SEQ ID NO: 193 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

193.

103. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 152 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 152, and a light chain constant region that comprises or consists of SEQ ID NO: 194 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

194.

104. The inducible IL-10 prodrug of any one of claims 1-11, 13-36, and 63-74, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 153 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 153, and a light chain constant region that comprises or consists of SEQ ID NO: 195 or an amino acid sequence that has at least 90% identity to SEQ ID NO:

195.

105. The inducible IL-10 prodrug of claim 65, wherein the antibody or antigen-binding fragment comprises an antibody light chain (VL + CL) comprising the amino acid sequence of any one of SEQ ID NOs: 127-167 and an antibody heavy chain Fab fragment (VH + CH1) comprising the amino acid sequence of any one of SEQ ID NOs: 168-194, and 500-513.

106. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises any one of Compounds 1-93 and 97-100 as shown in Table 10.

107. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 68, 72-76, 81-87, 93-104, 116-119, 123-126, and 514-567, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NOs: 69, 70, 71, 568-621.

108. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises Compound 1 (SEQ ID NO: 514 / SEQ ID NO: 568), Compound 2 (SEQ ID NO: 515 / SEQ ID NO: 569), Compound 3 (SEQ ID NO: 516 / SEQ ID NO: 570), Compound 4 (SEQ ID NO: 517 / SEQ ID NO: 571), Compound 5 (SEQ ID NO: 518 / SEQ ID NO: 572), Compound 6 192 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 (SEQ ID NO: 519 / SEQ ID NO: 573), Compound 7 (SEQ ID NO: 520 / SEQ ID NO: 574), Compound 8 (SEQ ID NO: 521 / SEQ ID NO: 575), Compound 9 (SEQ ID NO: 522 / SEQ ID NO: 576), Compound 10 (SEQ ID NO: 523 / SEQ ID NO: 577), Compound 11 (SEQ ID NO: 524 / SEQ ID NO: 578), Compound 12 (SEQ ID NO: 525 / SEQ ID NO: 579), Compound 13 (SEQ ID NO: 526 / SEQ ID NO: 580), Compound 14 (SEQ ID NO: 527 / SEQ ID NO: 581), Compound 15 (SEQ ID NO: 528 / SEQ ID NO: 582), Compound 16 (SEQ ID NO: 529 / SEQ ID NO: 583), Compound 17 (SEQ ID NO: 530 / SEQ ID NO: 584), Compound 18 (SEQ ID NO: 531 / SEQ ID NO: 585), Compound 19 (SEQ ID NO: 532 / SEQ ID NO: 586), Compound 20 (SEQ ID NO: 533 / SEQ ID NO: 587), Compound 21 (SEQ ID NO: 534 / SEQ ID NO: 588), Compound 22 (SEQ ID NO: 535 / SEQ ID NO: 589), Compound 22 (SEQ ID NO: 535 / SEQ ID NO: 589), Compound 23 (SEQ ID NO: 536 / SEQ ID NO: 590), Compound 24 (SEQ ID NO: 537 / SEQ ID NO: 591), Compound 25 (SEQ ID NO: 538 / SEQ ID NO: 592), Compound 26 (SEQ ID NO: 539 / SEQ ID NO: 593), Compound 27 (SEQ ID NO: 540 / SEQ ID NO: 594), Compound 28 (SEQ ID NO: 541 / SEQ ID NO: 595), Compound 29 (SEQ ID NO: 542 / SEQ ID NO: 596), Compound 30 (SEQ ID NO: 543 / SEQ ID NO: 597), Compound 31 (SEQ ID NO: 544 / SEQ ID NO: 598), Compound 32 (SEQ ID NO: 545 / SEQ ID NO: 599), Compound 33 (SEQ ID NO: 546 / SEQ ID NO: 600), Compound 34 (SEQ ID NO: 547 / SEQ ID NO: 601), Compound 35 (SEQ ID NO: 548 / SEQ ID NO: 602), Compound 36 (SEQ ID NO: 549 / SEQ ID NO: 603), Compound 37 (SEQ ID NO: 550 / SEQ ID NO: 604), Compound 38 (SEQ ID NO: 551 / SEQ ID NO: 605), Compound 39 (SEQ ID NO: 552 / SEQ ID NO: 606), Compound 40 (SEQ ID NO: 553 / SEQ ID NO: 607), Compound 41 (SEQ ID NO: 554 / SEQ ID NO: 608), Compound 42 (SEQ ID NO: 555 / SEQ ID NO: 609), Compound 43 (SEQ ID NO: 556 / SEQ ID NO: 610), Compound 44 (SEQ ID NO: 557 / SEQ ID NO: 611), Compound 45 (SEQ ID NO: 558 / SEQ ID NO: 612), Compound 46 (SEQ ID NO: 559 / SEQ ID NO: 613), Compound 47 (SEQ ID NO: 560 / SEQ ID NO: 614), Compound 48 (SEQ ID NO: 561 / SEQ ID NO: 615), Compound 49 (SEQ ID NO: 562 / SEQ ID NO: 616), Compound 50 (SEQ ID NO: 563 / SEQ ID NO: 617), Compound 51 (SEQ ID NO: 564 / SEQ ID NO: 618), Compound 52 (SEQ ID NO: 565 / SEQ ID NO: 619), Compound 53 (SEQ ID NO: 566 / SEQ ID NO: 620), or Compound 54 (SEQ ID NO: 567 / SEQ ID NO: 621).

109. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 531, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

585.

110. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid 193 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 sequence of SEQ ID NO: 532, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

586.

111. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 533, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

587.

112. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 534, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

588.

113. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 535, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

589.

114. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 536, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

590.

115. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 540, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

594.

116. The inducible IL-10 prodrug of any one of claims 1-36, 63-74, 84-91, and 93-105, wherein the inducible IL-10 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 542, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NO:

596. 194 \\4147-0268-4242 v1Attorney Docket No.: 761146.192320 117. An inducible IL-10 prodrug comprising an IL-10 polypeptide, a half-life extension element, an IL-10 blocking element and a protease cleavable linker, wherein the IL-10 blocking element is an antigen binding fragment of an antibody, wherein the antigen binding fragment comprises as separate components, at least an antigen-binding portion of an antibody light chain and at least an antigen-binding portion of a complementary antibody heavy chain, and the inducible IL-10 prodrug comprises: i. a first polypeptide comprising the IL-10 polypeptide, at least an antigen binding portion of an antibody light chain, and the half-life-extension element; wherein the IL-10 polypeptide and the antigen binding portion of the antibody light chain and / or half-life extension element are operably linked by the protease cleavable linker; wherein antigen binding portion of an antibody light chain comprises any one of SEQ ID NOs: 168-194 and 500-513; wherein the protease-cleavable peptide linker comprises any one of SEQ ID NOs: 5, 9-11, 14-35, 37-43, 46-49, 52-58, 105-108, 112-115, 195-201, 249 or 259; and ii. a second polypeptide comprising at least an antigen binding portion of an antibody heavy chain that is complementary to the light chain in the first polypeptide, together with said light chain forms an IL-10 binding site; wherein the antigen binding portion of an antibody heavy chain comprises any one of SEQ ID NOs: 514-567. 195 \\4147-0268-4242 v1

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