Inducible il-18 prodrugs

Abstract

Provided herein are IL- 18 polypeptide prodrugs comprising IL- 18, a half-life extension element, an IL- 18 blocking element and a protease cleavable linker. Also provided herein are pharmacal compositions thereof, as well as nucleic acids, recombinant expression vectors, host cells for making such polypeptide prodrugs. Also disclosed are methods of using the polypeptide prodrugs in the treatment of diseases, conditions and disorders.

Description

[0001] Attorney Docket No.: 761146.362320 INDUCIBLE IL-18 PRODRUGS [1] The present application claims the benefit of U.S. Provisional Application No.: 63 / 618,839, filed on January 8, 2024, and U.S. Provisional Application No.: 63 / 566,642, filed on March 18, 2024, U.S. Provisional Application No.: 63 / 575,107, filed on April 5, 2024, U.S. Provisional Application No.: 63 / 717,066, filed November 6, 2024, each of which are incorporated by reference in their entirety. 1. BACKGROUND [2] Interleukin-18 (IL-18) is a pro-inflammatory cytokine that plays a crucial role in the immune system. It is known for its ability to induce cell-mediated immunity following infections by various pathogens. IL-18 is involved in numerous immune responses, including the production of interferon- gamma (IFN-γ) in T-cells and natural killer cells. See, Ihim et al., (2022), Front Immunol., 13:919973. [3] IL-18 has potential as an anticancer drug, but its interactions with the inhibitory decoy receptor IL-18 binding protein (IL-18BP), which is frequently upregulated in tumors, has limited its clinical utility as an antitumor agent. See, Deckers et al., (2023), Nature, 1:286-3033. [4] Unfortunately, due to the biology of cytokines and the inability to effectively target and control their activity, cytokines have not achieved the hoped for clinical advantages in the treatment of tumors. This is exacerbated by the need to administer large quantities of cytokines (i.e., IL-18) in order to achieve the desired levels of cytokine at the intended site of cytokine action (e.g., a tumor microenvironment). [5] Inducible IL-12, IL-2, and interferon prodrugs have been described in International Publication Nos.: WO2019 / 222294, WO2019 / 222295, WO2019 / 222296, WO2021 / 097376. [6] There remains an unmet medical need for improved methods for treating cancer. 2. SUMMARY [7] The disclosure relates to inducible IL-18 prodrugs that contain at least one polypeptide chain, and can contain two or more polypeptides, if desired. The inducible IL-18 prodrug comprises an IL-18 polypeptide, an IL-18 blocking element, a half-life extension element, and a protease cleavable polypeptide linker. [8] Inducible IL-18 prodrugs of this disclosure have attenuated IL-18 receptor agonist activity and the circulating half-life is extended. The inducible IL-18 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-18 by noncovalently binding to the IL-18 and / or sterically blocking receptor binding. Upon cleavage of the protease cleavable linker a form of the IL-18 is released that is active (e.g., more active than the IL-18 polypeptide prodrug). Typically, the released IL-18 is at least 10 x more active than 1 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 the IL-18 polypeptide prodrug. Preferably, the released IL-18 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 10,000 x or more active than the inducible IL-18 prodrug. [9] The form of IL-18 that is released upon cleavage of the inducible IL-18 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-18 prodrug is extended, toxicity is reduced or eliminated because the agonist activity of the circulating inducible cytokine prodrug is attenuated and active IL-18 is targeted to the desired site of activity (e.g., tumor microenvironment).

[0010] The inducible IL-18 prodrug can comprise at least one of each of an IL-18 polypeptide [A], an IL-18 polypeptide that comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245, [D] a IL-18 blocking element, a half-life extension element [H], and a protease-cleavable polypeptide linker [L]. The IL-18 polypeptide and the IL-18 blocking element or the half-life extension element can be operably linked by the protease-cleavable polypeptide linker and the inducible IL-18 prodrug has attenuated IL-18 receptor activating activity. The IL-18 receptor activating activity of the inducible IL-18 prodrug is at least about 10X less than the IL-18 receptor activating activity of the polypeptide that contains the IL-18 polypeptide that is produced by cleavage of the protease cleavable linker.

[0011] The inducible IL-18 prodrug can comprise an IL-18 polypeptide [A], an IL-18 polypeptide that comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245, [H], a half-life extension element that comprises any one of SEQ ID NOs: 602-606 and 608-654, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-684, [D] a IL-18 blocking element, and a protease-cleavable polypeptide linker [L] that comprises any one of SEQ ID NOs: 354-379 or an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of 354-379.

[0012] The inducible IL-18 prodrug can have the formula:

[0013] [A]-[L1]-[H]-[L2]-[D];

[0014] [D]-[L2]-[H]-[L1]-[A];

[0015] [A]-[L1]-[D]-[L2]-[H];

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

[0017] [H]-[L1]-[A]-[L2’]-[D]; or

[0018] [D]-[L1]-[A]-[L2’]-[H].

[0019] [A] is an IL-18 polypeptide, [D] is an IL-18 blocking element, [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 [L2’] is a protease-cleavable polypeptide linker. L2 can be a protease-cleavable polypeptide linker. L1 or L2 or both L1 and L2 are cleaved by two or more different proteases. 2 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0020] The inducible IL-18 prodrug disclosed herein can comprise an antigen binding fragment of an antibody as the IL-18 blocking element. The antigen binding fragment of an antibody 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-18 prodrug can comprise: a first polypeptide comprising the IL-18 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-18 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 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-18 binding site.

[0021] The half-life life extension element can be a serum albumin binding domain, a serum albumin, transferrin, or immunoglobulin Fc, or fragment thereof. The half-life extension element is preferably a serum albumin binding domain. The serum albumin binding domain can be a single domain antibody fragment (sdAb). When the half-life extension element is a sdAb, the sdAb can comprise the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-654, an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-654.

[0022] IL-18 blocking domain can be a ligand-binding domain or fragment of a cognate receptor for the cytokine polypeptide, or an antibody or antigen-binding fragment of an antibody that binds to the cytokine polypeptide. The antibody or antigen-binding fragment is a single domain antibody, a Fab, or a scFv that binds the cytokine polypeptide. Preferably, the antibody or antigen-binding fragment is a Fab.

[0023] The blocking element can comprise an antibody light chain (VL + CL) comprising the amino acid sequence of SEQ ID NO: 172 or SEQ ID NO: 174, and an antibody heavy chain Fab fragment (VH + CH1) comprising the amino acid sequence of SEQ ID NO: 171 or SEQ ID NO: 173.

[0024] The blocking element can comprise an antibody light chain (VL + CL) comprising the amino acid sequence of SEQ ID NO: 172 or SEQ ID NO: 174, and antibody heavy chain Fab fragment (VH + CH1) comprising the amino acid sequence of SEQ ID NO: 171 or SEQ ID NO: 173.

[0025] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide 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 and 2) a second polypeptide that comprises or consists of SEQ ID NO: 172, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 172.

[0026] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 173, or an amino acid sequence 3 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 that has at least 95% identity to SEQ ID NO: 173 and 2) a second polypeptide that comprises or consists of SEQ ID NO: 174, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 174.

[0027] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises the amino acid sequence of SEQ ID NO: 318, and 2) a second polypeptide that comprises of SEQ ID NO: 319, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 319.

[0028] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises the amino acid sequence of SEQ ID NO: 320, and 2) a second polypeptide that comprises SEQ ID NO: 321.

[0029] The blocking element can comprise 1) a first polypeptide chain that comprises the amino acid sequence of SEQ ID NO: 322, and 2) a second polypeptide that comprises SEQ ID NO: 323.

[0030] The blocking element disclosed herein can be Blocking Element 1 (SEQ ID NO: 1033 / SEQ ID NO: 1058), Blocking Element 2 (SEQ ID NO: 1034 / SEQ ID NO: 1059), Blocking Element 3 (SEQ ID NO: 1035 / SEQ ID NO: 1060), Blocking Element 4 (SEQ ID NO: 1036 / SEQ ID NO: 1061), Blocking Element 5 (SEQ ID NO: 1037 / SEQ ID NO: 1062), Blocking Element 6 (SEQ ID NO: 1038 / SEQ ID NO: 1063), Blocking Element 7 (SEQ ID NO: 1039 / SEQ ID NO: 1064), Blocking Element 8 (SEQ ID NO: 1040 / SEQ ID NO: 1065), Blocking Element 9 (SEQ ID NO: 1041 / SEQ ID NO: 1066), Blocking Element 10 (SEQ ID NO: 1042 / SEQ ID NO: 1067), Blocking Element 11 (SEQ ID NO: 1043 / SEQ ID NO: 1068), Blocking Element 12 (SEQ ID NO: 1044 / SEQ ID NO: 1069), Blocking Element 13 (SEQ ID NO: 1045 / SEQ ID NO: 1070), Blocking Element 14 (SEQ ID NO: 1046 / SEQ ID NO: 1071), Blocking Element 15 (SEQ ID NO: 1047 / SEQ ID NO: 1072), Blocking Element 16 (SEQ ID NO: 1048 / SEQ ID NO: 1073), Blocking Element 17 (SEQ ID NO: 1049 / SEQ ID NO: 1074), Blocking Element 18 (SEQ ID NO: 1050 / SEQ ID NO: 1075), Blocking Element 19 (SEQ ID NO: 1051 / SEQ ID NO: 1076), Blocking Element 20 (SEQ ID NO: 1052 / SEQ ID NO: 1077), Blocking Element 21 (SEQ ID NO: 1053 / SEQ ID NO: 1078), Blocking Element 22 (SEQ ID NO: 1054 / SEQ ID NO: 1079), Blocking Element 23 (SEQ ID NO: 1055 / SEQ ID NO: 1080), Blocking Element 24 (SEQ ID NO: 1056 / SEQ ID NO: 1081), or Blocking Element 25 (SEQ ID NO: 1057 / SEQ ID NO: 1082).

[0031] The IL-18 blocking element disclosed herein inhibits activation of the cytokine polypeptide receptor by the inducible cytokine prodrug.

[0032] The protease cleavable linker can comprise a sequence that is capable of being cleaved by a protease selected from kallikrein, thrombin, chymase, carboxypeptidase A, cathepsin, elastase, PR-3, granzyme M, a calpain, a matrix metalloproteinase (MMP), an ADAM, a FAP, a plasminogen activator, a caspase, a tryptase, or a tumor protease. The protease cleavable linker can be selected from cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, or cathepsin G. The protease cleavable linker can be selected from matrix metalloprotease (MMP) is MMP1, MMP2, MMP3, MMP8, MMP9, 4 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 MMP10, MMP11, MMP12, MMP13, or MMP14. The protease cleavable linker can comprise at least two sequences that are independently capable of being cleaved by a protease. The protease cleavable linker can comprise a synthetic sequence.

[0033] The protease cleavable linker can comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 354-379 or an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of 354-379. The protease cleavable linker can comprise an amino acid sequence of SEQ ID NO: 354. The protease cleavable linker can comprise an amino acid sequence of SEQ ID NO: 357.

[0034] The inducible IL-18 prodrug disclosed herein can be Compound 1 (SEQ ID NO: 175 / SEQ ID NO: 187), Compound 2 (SEQ ID NO: 176 / SEQ ID NO: 187), Compound 3 (SEQ ID NO: 177 / SEQ ID NO: 187), Compound 4 (SEQ ID NO: 178 / SEQ ID NO: 187), Compound 5 (SEQ ID NO: 179 / SEQ ID NO: 187), Compound 6 (SEQ ID NO: 180 / SEQ ID NO: 187), Compound 7 (SEQ ID NO: 181 / SEQ ID NO: 187), Compound 8 (SEQ ID NO: 182 / SEQ ID NO: 187), Compound 9 (SEQ ID NO: 183 / SEQ ID NO: 188), Compound 10 (SEQ ID NO: 184 / SEQ ID NO: 188), Compound 11 (SEQ ID NO: 185 / SEQ ID NO: 188), or Compound 12 (SEQ ID NO: 186 / SEQ ID NO: 188), Compound 13 (SEQ ID NO: 246 / SEQ ID NO: 311), Compound 14 (SEQ ID NO: 247 / SEQ ID NO: 312), Compound 15 (SEQ ID NO: 248 / SEQ ID NO: 312), Compound 16 (SEQ ID NO: 249 / SEQ ID NO: 314), Compound 17 (SEQ ID NO: 250 / SEQ ID NO: 313), Compound 18 (SEQ ID NO: 251 / SEQ ID NO: 313), Compound 19 (SEQ ID NO: 252 / SEQ ID NO: 313), Compound 20 (SEQ ID NO: 253 / SEQ ID NO: 313), Compound 21 (SEQ ID NO: 254 / SEQ ID NO: 313), Compound 22 (SEQ ID NO: 255 / SEQ ID NO: 311), Compound 23 (SEQ ID NO: 256 / SEQ ID NO: 311), Compound 24 (SEQ ID NO: 257 / SEQ ID NO: 313), Compound 25 (SEQ ID NO: 258 / SEQ ID NO: 313), Compound 26 (SEQ ID NO: 259 / SEQ ID NO: 314), Compound 27 (SEQ ID NO: 260 / SEQ ID NO: 314), Compound 28 (SEQ ID NO: 261 / SEQ ID NO: 314), Compound 29 (SEQ ID NO: 262 / SEQ ID NO: 314), Compound 30 (SEQ ID NO: 263 / SEQ ID NO: 313), Compound 31 (SEQ ID NO: 264 / SEQ ID NO: 313), Compound 32 (SEQ ID NO: 265 / SEQ ID NO: 315), Compound 33 (SEQ ID NO: 266 / SEQ ID NO: 315), Compound 34 (SEQ ID NO: 267 / SEQ ID NO: 315), Compound 35 (SEQ ID NO: 268 / SEQ ID NO: 315), Compound 36 (SEQ ID NO: 269 / SEQ ID NO: 316), Compound 37 (SEQ ID NO: 270 / SEQ ID NO: 316), Compound 38 (SEQ ID NO: 271 / SEQ ID NO: 316), Compound 39 (SEQ ID NO: 272 / SEQ ID NO: 316), Compound 40 (SEQ ID NO: 273 / SEQ ID NO: 315), Compound 41 (SEQ ID NO: 274 / SEQ ID NO: 315), Compound 42 (SEQ ID NO: 275 / SEQ ID NO: 315), Compound 43 (SEQ ID NO: 276 / SEQ ID NO: 315), Compound 44 (SEQ ID NO: 277 / SEQ ID NO: 315), Compound 45 (SEQ ID NO: 278 / SEQ ID NO: 315), Compound 46 (SEQ ID NO: 279 / SEQ ID NO: 315), Compound 47 (SEQ ID NO: 280 / SEQ ID NO: 315), Compound 48 (SEQ ID NO: 281 / SEQ ID NO: 317), Compound 49 (SEQ ID NO: 282 / SEQ ID NO: 317), Compound 50 (SEQ ID NO: 283 / SEQ ID NO: 317), Compound 51 (SEQ ID NO: 284 / SEQ 5 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 ID NO: 317), Compound 52 (SEQ ID NO: 285 / SEQ ID NO: 317), Compound 53 (SEQ ID NO: 286 / SEQ ID NO: 317), Compound 54 (SEQ ID NO: 287 / SEQ ID NO: 313), Compound 55 (SEQ ID NO: 288 / SEQ ID NO: 313), Compound 56 (SEQ ID NO: 289 / SEQ ID NO: 313), Compound 57 (SEQ ID NO: 290 / SEQ ID NO: 313), Compound 58 (SEQ ID NO: 291 / SEQ ID NO: 313), Compound 59 (SEQ ID NO: 292 / SEQ ID NO: 313), Compound 60 (SEQ ID NO: 293 / SEQ ID NO: 313), Compound 61 (SEQ ID NO: 294 / SEQ ID NO: 313), Compound 62 (SEQ ID NO: 295 / SEQ ID NO: 314), Compound 63 (SEQ ID NO: 296 / SEQ ID NO: 314), Compound 64 (SEQ ID NO: 297 / SEQ ID NO: 314), Compound 65 (SEQ ID NO: 298 / SEQ ID NO: 314), Compound 66 (SEQ ID NO: 299 / SEQ ID NO: 314), Compound 67 (SEQ ID NO: 300 / SEQ ID NO: 314), Compound 68 (SEQ ID NO: 301 / SEQ ID NO: 316), Compound 69 (SEQ ID NO: 302 / SEQ ID NO: 316), Compound 70 (SEQ ID NO: 303 / SEQ ID NO: 316), Compound 71 (SEQ ID NO: 304 / SEQ ID NO: 316), Compound 72 (SEQ ID NO: 305 / SEQ ID NO: 311), Compound 73 (SEQ ID NO: 306 / SEQ ID NO: 315), Compound 74 (SEQ ID NO: 307 / SEQ ID NO: 315), Compound 75 (SEQ ID NO: 308 / SEQ ID NO: 316), Compound 76 (SEQ ID NO: 309 / SEQ ID NO: 316), or Compound 77 (SEQ ID NO: 310 / SEQ ID NO: 313).

[0035] The inducible IL-18 prodrugs disclosed herein can be Compound 78, Compound 79, Compound 80, Compound 81, Compound 82, Compound 83, Compound 84, Compound 85, Compound 86, Compound 87, Compound 88, Compound 89, Compound 90, Compound 91, Compound 92, Compound 93, Compound 94, Compound 95, Compound 96, Compound 97, Compound 98, Compound 99, Compound 100, Compound 101, Compound 102, Compound 103, Compound 104, Compound 105, Compound 106, Compound 107, Compound 108, Compound 109, Compound 110, Compound 111, Compound 112, Compound 113, Compound 114, Compound 115, Compound 116, Compound 117, Compound 118, Compound 119, Compound 120, Compound 121, Compound 122, Compound 123, Compound 124, Compound 125, Compound 126, Compound 127, Compound 128, Compound 129, Compound 130, Compound 131, Compound 132, Compound 133, Compound 134, Compound 134, Compound 135, Compound 136, Compound 137, Compound 138, Compound 139, Compound 140, Compound 141, Compound 142, Compound 143, Compound 144, Compound 145, Compound 146, Compound 147, Compound 148, Compound 149, Compound 150, Compound 151, Compound 152, Compound 153, Compound 154, Compound 155, Compound 156, Compound 157, Compound 158, Compound 159, Compound 160, Compound 161, Compound 162, Compound 163, Compound 164, Compound 165, Compound 166, Compound 167, Compound 168, Compound 169, Compound 170, Compound 171, Compound 172, Compound 173, Compound 174, Compound 175, Compound 176, Compound 177, Compound 178, Compound 179, Compound 180, Compound 181, Compound 182, Compound 183, Compound 184, Compound 184, Compound 185, Compound 186, Compound 187, Compound 188, Compound 189, Compound 190, Compound 191, Compound 192, Compound 193, 6 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 194, Compound 195, Compound 196, Compound 197, Compound 198, Compound 199, Compound 200, Compound 201, Compound 202, Compound 203, Compound 204, Compound 205, Compound 206, Compound 207, Compound 208, Compound 209, Compound 210, Compound 211, Compound 212, Compound 213, Compound 214, Compound 215, Compound 216, Compound 217, Compound 218, Compound 219, Compound 220, Compound 221, Compound 222, Compound 223, Compound 224, Compound 225, Compound 226, Compound 227, Compound 228, Compound 229, Compound 230, Compound 231, Compound 232, Compound 233, Compound 234, Compound 235, Compound 236, Compound 237, Compound 238, Compound 239, Compound 240, Compound 241, Compound 242, Compound 243, Compound 244, Compound 245, Compound 246, Compound 247, Compound 248, Compound 249, Compound 250, Compound 251, Compound 252, Compound 253, Compound 254, Compound 255, Compound 256, Compound 257, Compound 258, Compound 259, Compound 260, Compound 261, Compound 262, Compound 263, Compound 264, Compound 265, Compound 266, Compound 267, Compound 268, Compound 269, Compound 270, Compound 271, Compound 272, Compound 273, Compound 274, Compound 275, Compound 276, Compound 277, Compound 278, or Compound 279.

[0036] The inducible IL-18 prodrug disclosed herein can comprise at least one of each of: a) an IL-18 polypeptide [A], wherein the IL-18 polypeptide comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245; b) a half-life extension element [H], wherein the half-life extension element comprises any one of SEQ ID NOs: 602-606 and 608-654, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-684; c) an IL-18 blocking element [D]; and d) a protease cleavable linker [L], wherein the protease cleavable linker comprises any one of SEQ ID NOs: 355-379 or 426 or an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of 355-379 or 426. The IL-18 polypeptide and the IL-18 blocking element or the half-life extension element are operably linked by the protease-cleavable polypeptide linker and the inducible IL-18 prodrug has attenuated IL-18 receptor activating activity, wherein the IL-18 receptor activating activity of the inducible IL-18 prodrug is at least about 10X less than the IL-18 receptor activating activity of the polypeptide that contains the IL-18 polypeptide that is produced by cleavage of the protease cleavable linker.

[0037] The inducible IL-18 prodrug described herein can comprise an IL-18 polypeptide, a half-life extension element, an IL-18 blocking element and a protease cleavable linker, wherein the IL-18 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-18 prodrug comprises: i) a first polypeptide comprising the IL-18 polypeptide, wherein the IL-18 polypeptide 7 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245; a half-life extension element, wherein the half-life extension element comprises any one of SEQ ID NOs: 602-606 and 608-654, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-654; an antigen binding portion of an antibody heavy chain, wherein the antibody binding portion of the heavy chain comprises any one of SEQ ID NOs: 171, 173, 318, 320, 331, 322, 329, 330, 337, 339, 353, 354, 655-660, 662, 664, 733-736, 738-742, 820-823, 875-877, 920, 966-969, 1005, 1006, 1010, or 1011;wherein the IL-18 polypeptide and the antigen binding portion of the antibody heavy chain and / or half-life extension element are operably linked by the protease cleavable linker, wherein the protease cleavable linker comprises any one of SEQ ID NOs: 354-379; and 2) a second polypeptide comprising at least an antigen binding portion of an antibody light chain that is complementary to the heavy chain in the first polypeptide and together with said light chain forms an IL-18 binding site, wherein the antibody light chain comprises any one of SEQ ID NOs: 172, 174, 187, 188, 311, 312, 314- 316, 319, 321, 323, 815, 816, 862-872, 1012-1016, or 1019.

[0038] The disclosure also relates to a nucleic acid encoding the inducible IL-18 prodrugs. The nucleic acid can comprise a circular vector. The nucleic acid can comprise DNA. The nucleic acid can comprise RNA. Disclosed herein are methods of making the pharmaceutical composition comprising culturing the host cell under suitable conditions for expression and collection of the inducible IL-18 prodrug. Disclosed herein is a nucleic acid composition comprising one or more nucleic acid sequences encoding an inducible IL-18 prodrug. The inducible IL-18 prodrug comprises at least one of each of an IL-18 polypeptide [A]; an IL-18 blocking element [D]; a half-life extension element [H]; and a protease- cleavable polypeptide linker [L]. The IL-18 polypeptide and the IL-18 blocking element or the half-life extension element are operably linked by the protease-cleavable polypeptide linker and the inducible IL- 18 prodrug has attenuated IL-18 receptor activity.

[0039] The nucleic composition can comprise one or more nucleic acid sequences encoding an inducible IL-18 prodrug containing two polypeptide chains. The inducible IL-18 prodrug can comprise an antigen binding fragment of an antibody as the IL-18 blocking element. The antigen binding fragment of an antibody 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- 18 prodrug can comprise: a first polypeptide comprising the IL-18 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-18 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 second polypeptide comprising at least an antigen binding portion of an antibody heavy chain that is complementary to the light chain in the first 8 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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-18 binding site.

[0040] The disclosure also relates to therapeutic methods that include administering to a subject in need thereof an effective amount of an inducible IL-18 prodrug, nucleic acid that encodes the inducible IL-18 prodrug, vector or host cells that contain such a nucleic acid, and pharmaceutical compositions of any of the foregoing. Typically, the subject has, or is at risk of developing cancer, a proliferative disease, a tumorous disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, a graft-versus-host disease, or a host-versus-graft disease. The methods disclosed herein are particularly suitable for treating cancer. The inducible IL-18 prodrug can be administered intravenously. 3. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] 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 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:

[0042] FIGs.1A-1P are schematics of inducible IL-18 prodrugs.

[0043] FIG.2A is a graph showing the anti-tumor activity of WW50941 / WW50632 (an inducible IL-18 prodrug containing binding protein resistant IL-18 (“BPR”) as compared to WW50629 / WW50632 (an inducible IL-18 prodrug containing wild-type IL-18) in mice bearing MC38 tumors. It shows the average tumor volume over time in mice treated with the doses documented in the figure legend. FIGs.2B-2D are graphs showing individual spider plots on mice treated with the inducible BPR IL-18 prodrug, inducible WT IL-18 prodrug, and PBS. CR = complete regression.

[0044] FIGs.3A, 3D, 3G, 3J, 3M, 3P, 3S, and 3V are graphs depicting results from HEK-Blue IL-18 reporter assay for inducible IL-18 prodrugs. Analysis was performed on quantification of Secreted Alkaline Phosphate (SEAP) activity using the reagent QUANTI-Blue (InvivoGen). FIGs.3B, 3E, 3H, 3K, 3N, 3Q, 3T, and 3W are graphs showing the activity of IL-18 proteins in an IL-18 PBMC assay. EC50 values for each protein are shown. Analysis was performed based on quantification of IFNγ in cell supernatants. Circles represent various recombinant IL-18 cytokines representative of the inducible IL-18 prodrugs, squares represent the intact inducible IL-18 prodrugs, and triangles represent the cleaved inducible IL-18 prodrugs.

[0045] FIGs.3C, 3F, 3I, 3L, 3O, 3R, 3U, and 3X show the results of protein cleavage assays of inducible IL-18 prodrugs run on an SDS-PAGE gel. The inducible IL-18 prodrugs were run in both the intact and cleaved (elastase cleaved )form. All samples were run under non-reducing conditions. 9 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0046] FIGs.4A-4O are graphs depicting the activity of inducible IL-18 prodrugs in an IL-18 human PBMC assay. Analysis was performed based on quantification of IFNγ in cell supernatants, and EC50 values for each protein are shown. Circles represent various recombinant IL-18 cytokines representative of the IL-18 cytokine variant in the inducible IL-18 prodrugs, squares represent the intact inducible IL-18 prodrugs, and triangles represent the cleaved inducible IL-18 prodrugs.

[0047] FIGs.5A-5E are graphs showing results of analyzing inducible IL-18 prodrug, WW50629 / WW50632, in a syngeneic MC38 mouse tumor model. FIG.5A shows average tumor volume over time in mice treated with 37.5 µg, 75 µg, 150 µg, and 300 µg of WW50629 / WW50632. FIGs.5B- 5E shows tumor volume over study days in mice for each dose of WW50629 / WW50632 at 37.5 µg (FIG. 5B), 75 µg (FIG.5C), 150 µg (FIG.5D), and 300 µg (FIG.5E). FIG.5F is a graph showing body weight averages for the animals treated in FIGs.5A-5E. Each line in the plot represents the body weight average of the group over time.

[0048] FIGs.6A-6E are graphs showing results of analyzing inducible IL-18 prodrug, WW50938 / WW50632, in a syngeneic MC38 mouse tumor model. FIG.6A shows average tumor volume over time in mice treated with 37.5 µg, 75 µg, 150 µg, and 300 µg of WW50938 / WW50632. FIGs.6B- 6E shows tumor volume over study days in mice for each dose of WW50938 / WW50632 at 37.5 µg (FIG. 6B), 75 µg (FIG.6C), 150 µg (FIG.6D), and 300 µg (FIG.6E). FIG.6F is a graph showing body weight averages for the animals treated in FIGs.6A-6E. Each line in the plot represents the body weight average of the group over time.

[0049] FIGs.7A-7E are graphs showing results of analyzing inducible IL-18 prodrug, WW50941 / WW50632, in a syngeneic MC38 mouse tumor model. FIG.7A shows average tumor volume over time in mice treated with 37.5 µg, 75 µg, 150 µg, and 300 µg of WW50941 / WW50632. FIGs.7B- 7E shows tumor volume over study days in mice for each dose of WW50941 / WW50632 at 37.5 µg (FIG. 7B), 75 µg (FIG.7C), 150 µg (FIG.7D), and 300 µg (FIG.7E). FIG.7F is a graph showing body weight averages for the animals treated in FIGs.7A-7E. Each line in the plot represents the body weight average of the group over time.

[0050] FIG.8 is a table showing the number of deaths of mice treated with the inducible IL-18 prodrugs in FIGs.5A-5E, 6A-6E, and 7A-7E.

[0051] FIGs.9A-9C are graphs showing the activity of IL-18 proteins in an IL-18 murine splenocyte assay in media without human serum albumin. FIG.9A shows the activity of WW50295 (a wild-type murine IL-18 protein) in comparison to the activity of either intact or cleaved (activated) WW50482 / WW50487 (an inducible IL-18 prodrug containing wild-type mIL-18 protein). FIG.9B shows the activity of WW50249, a murine IL-18 protein that does not bind IL-18BP (binding protein resistant IL-18, (BPR IL-18)) in comparison to the activity of either intact or cleaved (activated) 10 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 WW50480 / WW50487 (an inducible IL-18 prodrug containing BPR mIL-18 protein). FIG.9C shows the activity of WW50249 (a BPR mIL-18 protein) in comparison to the activity of either intact or cleaved (activated) WW50345 (a cleavable half-life extended molecule containing a BPR mIL-18 protein). Circles represent the recombinant protein, while squares depict the activity of intact molecules and triangles depict the activity of the cleaved molecules. EC50 values for each protein are shown in the associated table, and biological replicates are documented in Tables 8 and 9. Analysis was performed based on the quantification of cell proliferation using Cell Titer Glo. Results confirm that both WW50482 / WW50487 and WW50480 / WW50487 have inducible activity in murine splenocytes.

[0052] FIGs.10A-10C are graphs showing the activity of IL-18 proteins in an IL-18 murine splenocyte assay in media with 0.5% weight / volume human serum albumin added. FIG.10A shows the activity of WW50295 (a wild-type murine IL-18 protein) in comparison to the activity of either intact or cleaved (activated) WW50482 / WW50487 (an inducible IL-18 prodrug containing wild-type mIL-18 protein). FIG.10B shows the activity of WW50249 (a BPR murine IL-18 protein) in comparison to the activity of either intact or cleaved (activated) WW50480 / WW50487 (an inducible IL-18 prodrug containing a BPR mIL-18 protein). FIG.10C shows the activity of WW50249 (a BPR murine IL-18 protein) in comparison to the activity of either intact or cleaved (activated) WW50345 (a cleavable half-life extended molecule containing a BPR mIL-18 protein). Circles represent the recombinant protein, while squares depict the activity of intact molecules and triangles depict the activity of the cleaved molecules. EC50 values for each protein are shown in the associated table, and biological replicates are documented in Tables 8 and 9. Analysis was performed based on the quantification of cell proliferation using Cell Titer Glo. Results confirm that WW50482 / WW50487, WW50480 / WW50487, and WW50345 have inducible activity in murine splenocytes.

[0053] FIG.11A is a graph showing body weight over time in mice administered BPR mIL-18 (WW50247) administered at 20 µg, 100 µg, 160 µg on a 5 / 2 / 5 / 2 dosing regimen and 100 µg bi-weekly. FIG.11B ia a graph showing body weight over time in mice administered mIL-18-anti-HSA (WW50159) at 33.5µg, inducible mIL-18 prodrug (WW50226) administered at 75.6µg, BPR mIL-18-HLE (WW50195) administered at 33.5µg and inducible BPR mIL-18 prodrug (WW50220) administered at 75.6 µg on a biwk x 2 dosing regimen. BPR mIL18, administered at 20µg on a 5 / 2 / 5 / 2 dosing regimen, showed a minimally efficacious tumor growth delay, while BPR mIL-18-HLE (WW50195) administered at 33.5µg on a biwk x 2 dosing regimen, showed tumor control. FIGs.11C shows tumor volume over time in inducible mIL-18 prodrug and inducible BPR mIL-18 prodrug. FIGs.11D shows tumor volume over time in inducible mIL-18 BPR and BPR mIL-18. FIGs.11E shows tumor volume over time BPR mIL-18-HLE. FIGs.11F shows tumor volume over time in inducible mIL-18 prodrug and inducible BPR mIL-18 prodrug. FIG.11G shows the effects on tumor growth delay of for equimolar amounts (total drug 11 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 dosed over 2 week dosing regimen) for BPR mIL-18 (WW50247) administered at 20µg on a 5 / 2 / 5 / 2 dosing regimen, BPR mIL-18-inducible prodrug (WW50226) administered at 76.6 µg on a biwk x 2 dosing regimen. BPR mIL-18-HLE (WW50195) at 33.5 µg on a biwk x 2, and BPR mIL-18 prodrug at 76.6 µg on a biwk x 2. Only BPR mIL-18-HLE (WW50195) administered at 33.5 µg on a biwk x 2 dosing regimen, showed tumor control.

[0054] FIG.11H shows the effects on tumor growth delay of for equimolar amounts (total drug dosed over 2 week dosing regimen) for BPR mIL-18 (WW50247) administered at 100µg on a 5 / 2 / 5 / 2 dosing regimen, and BPR mIL-18-HLE (WW50195) administered at 168 µg on a biwk x 2 dosing regimen. Only BPR mIL-18-HLE (WW50195) administered at 168 µg on a biwk x 2 dosing regimen, showed tumor control.

[0055] FIGs.12A-12D are graphs showing results of comparing inducible BPR mIL-18 prodrug (WW50480 / WW50487) to the inducible WT mIL-18 prodrug (WW50482 / WW50487) for efficacy and tolerability in the MC38 mouse model. FIG.12A shows tumor volume over time for inducible BPR mIL- 18 prodrug (WW50480 / WW50487) relative to vehicle. FIG.12B shows tolerability, as measured by weight loss for inducible BPR mIL-18 prodrug (WW50480 / WW50487). Treatment related deaths are noted. All doses showed efficacy as observed by tumor growth inhibition and regression. Four animals died in the 110µg dose group, 8 animals died in the 330µg dose group, and 8 animals died in the 1000µg dose group. FIG.12C shows tumor volume over time for inducible WT IL-18 prodrug (WW50482 / WW500487) relative to vehicle. FIG.12D shows tolerability, as measured by weight loss for the inducible WT mIL-18 prodrug (WW50482 / WW50487). Treatment related deaths are noted. The 330 µg and 1000 µg doses showed efficacy as observed by tumor growth inhibition and regression. Weight loss was evident in the 1000µg dose level. Three animals died in the 110µg dose group, 1 animal died in the 330µg dose group, and 6 animals died in the 1000µg dose group.

[0056] FIGs.13A, 13C, 13E, 13G are graphs showing tumor volume over time for BPR mIL-18 HLE (WW50217) (FIG.13A), inducible BPR mIL-18 prodrug (WW50481 / WW50487) (FIG.13C), inducible BPR mIL-18 prodrug (WW50480 / WW50487) (FIG.13E), and inducible wild-type mIL-18 prodrug (WW50482 / WW50487) (FIG.13G). FIG.s 13B, 13D, and 13F are graphs showing shows tolerability, as measured by weight loss for BPR mIL-18 HLE (WW50217) (FIG.13B), inducible BPR mIL-18 prodrug (WW50481 / WW50487) (FIG.13D), inducible BPR mIL-18 prodrug (WW50480 / WW50487) (FIG.13F), and inducible wild-type mIL-18 prodrug (WW50482 / WW50487) (FIG.13H).

[0057] FIGs.14A-14C are graphs showing serum concentration over time relationship for inducible BPR mIL-18 prodrug (at 50 µg) compared to inducible WT mIL-18 prodrug (300 µg). FIG.14A shows total serum concentration over time. There was a 2-fold reduction in T1 / 2 of WT compared to inducible BPR mIL-18 prodrug. FIG.14B is a graph showing free IL-18 measured in the serum at 24h, 72h, and 12 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 120h after dosing. Peak exposure of ~8nM and ~34nM respectively was detected at 24 hours for inducible BPR mIL-18 prodrug and inducible WT prodrug, respectively, which is proportional to the dose administered. FIG.14C is a graph showing serum IL-18:IL-18BP measured at 24h, 72h, and 120h after dosing. No IL-18:IL-18BP complex was detected in the inducible BPR mIL-18 prodrug dosed animals, whereas the IL-18:IL-18BP complex was detected in the inducible WT mIL-18 prodrug dosed animals which is first seen at ~5nM at 24 hours, peaks at 22.5nM at 72 hours, and detected at 11nM at 120 hours.

[0058] FIGs.15A and 15B are graphs showing serum IFNγ levels (FIG.15A) and serum IL-18BP levels (FIG.15B) after dosing with vehicle, inducible BPR mIL-18 prodrug, or inducible WT mIL-18 prodrug. Serum IFNγ was similar (~200 pg / mL) for both inducible BPR mIL-18 prodrug and inducible WT mIL18 prodrug at 24 hours. At 72 hours serum IFNγ increased to approximately 450pg / mL in inducible BPR mIL-18 prodrug treated animals and drops to barely detectable levels in the inducible WT mIL-18 prodrug treated animals. High levels (approximately 245pg / mL) were detectable at 120 hours in the inducible WT mIL-18 prodrug treated animals. Serum IL-18BP upregulation was higher in the inducible BPR mIL-18 prodrug treated animals which peaked at approximately 78 ng / mL at 120 hours compared to inducible WT mIL-18 prodrug which peaked at 23 ng / mL at 96 hours.

[0059] FIGs.16A-16D are graphs showing the efficacy of BPR IL-18 (WW50249) (FIG.16A), half-life extended (HLE) BPR IL-18 (non-cleavable, WW50217 (FIG.16B) and cleavable, WW50345 (FIG.16C), respectively), and inducible BPR IL-18 prodrug (WW50480 / 50487) as determined by tumor volume in the MC38 mouse model. Treatment days are shown with upward triangles on the x-axis.

[0060] FIGs.17A-17D are graphs showing the tolerability, as determined by body-weight changes, of BPR IL-18 (WW50249) (FIG.17A), BPR IL-18 HLE constructs (non-cleavable and cleavable, WW50217 (FIG.17B) and WW50345 (FIG.17C) respectively), and inducible BPR IL-18 prodrug (WW50480 / 50487) (FIG.17D) as determined by body weight changes in the MC38 mouse model. Treatment days are shown with upward triangles on the x-axis.

[0061] FIGs.18A and 18C are graphs showing anti-tumor activity of WW50480 / 50487 (an inducible IL-18 prodrug containing binding protein resistant (BPR) IL-18 in mice bearing MC38 tumors implanted with 50% Matrigel (FIG.18A) and in mice bearing MC38 tumors implanted in the absence of Matrigel (FIG.18C). It shows the average tumor volume over time in mice treated with the doses and complete responses (CRs) in the figure legend. FIGs.18B and 18D are graphs showing the change in body weight of MC38 tumor bearing animals treated with inducible IL-18 prodrug, WW50480 / 50487, in mice bearing MC38 tumors implanted with 50% Matrigel (FIG.18B) and in mice bearing MC38 tumors implanted in the absence of Matrigel (FIG.18D). Treatment related deaths are documented in the figure legend.

[0062] FIGs.19A-19F are graphs showing the levels of IL-18 binding protein (BP) (FIGs.19A and 19D), IFNγ (FIGs.19B and 19E), and IL-5 (FIGs.19C and 19F) measured in the tumor (FIGs.19A-19C), 13 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 or plasma (FIGs.19D-19F) of tumor bearing animals after treatment with an inducible IL-18 prodrug containing binding protein resistant (BPR) IL-18) at 24 hrs and 72hrs. The data show that treatment with WW50480 / 50487 increases the amount of IL-18BP and IFNγ found in the tumor 24 hours after dosing, and increases the amount of IL-18BP plasma at both timepoints, consistent with ongoing IL-18 signaling in vivo. Statistical analysis was conducted using 2-way ANOVA with Tukey’s multiple comparisons test. Significance is *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0063] FIGs.20A-20E are graphs showing levels of additional cytokines in the plasma at 24 hrs and 72 hrs after treatment with an inducible BPR IL-18 prodrug, WW50480 / WW50487. The data show that WW50480 / 50487 treatment increased the levels of levels of TNF (FIG.20A), IL-10 (FIG.20B), IL-4 (FIG.20C), IL-6 (FIG.20D) and KC / Gro (FIG.20E) in the plasma. Statistical analysis was conducted using conducted using 2-way ANOVA with Tukey’s multiple comparisons test. Significance is *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0064] FIGs.21A-21C are graphs showing the anti-tumor activity of WW50249 (a recombinant BPR mIL-18 cytokine) (FIG.21A), WW50345 (a recombinant BPR mIL-18 cytokine linked in a cleavable manner to a half-life extension domain) (FIG.21B), or WW50480 / 50487 (an inducible mIL-18 prodrug containing a BPR mIL-18 cytokine) (FIG.21C) in mice bearing EMT6 tumors. It shows average tumor volume over time in mice treated with the doses documented in the figure legend. In all three graphs, the vehicle treated animals are shown as black circles. The ascending dose levels are represented by the lines with square symbol (lowest dose), the upwards pointing triangle, the diamond, and in some cases, a downward pointing triangle. The specific doses are documented in each figure legend next to their representative symbol. The data show that the growth in tumor volume was minimally inhibited by dosing with WW50249, while the two highest doses of WW50345 inhibited tumor growth. In contrast, tumor growth was inhibited in all except the lowest dosing level of mice treated with WW50480 / 50487.

[0065] FIGs.22A-22L are graphs showing a series of spider plots showing activity of fusion proteins in an EMT6 mouse model corresponding to the data shown in FIGs.21A-21C. Each line in the plots is the tumor volume over time for a single mouse.

[0066] FIGs.23A-23E are graphs showing the efficacy of inducible IL-18 prodrugs in the MC38 model. FIG.23A is a graph showing the anti-tumor activity of WW50480 / 50487 (an inducible IL-18 prodrug containing binding protein resistant IL-18) or WW50482 / 50487 (an inducible IL-18 prodrug containing wildtype IL-18) in mice bearing MC38 tumors. It shows the average tumor volume over time in mice treated at doses 3 µg and 300 µg, respectively. FIG.23B is a graph showing the percent change in body weight of MC38 tumor bearing animals treated with inducible IL-18 prodrugs. FIG.23C-23E are spider plots showing tumor growth of individual animals. The data show that the growth in tumor volume was inhibited by both inducible IL-18 prodrugs. The anti-tumor activity of WW50480 / 50487 (BPR) was 14 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 better than that of WW50482 / 50487 (WT). WW50480 / 50487 (BPR) was well tolerated by tumor bearing mice, whereas treatment with WW50482 / 50487 resulted in the death of 2 / 8 animals.

[0067] FIGs.24A-24C are graphs showing the percent of MC38 tumor infiltrating immune cells that were either CD8+ T cells (FIGs.24A and 24C) or M2 macrophages (FIGs.24B and 24D) on Day 5 (FIGs.24A and 24B) or Day 11 (FIGs.24C and 24D) post treatment with WW50480 / WW50487 (BPR prodrug) or WW50482 / WW50487 (WT prodrug). The data show that treatment with inducible IL-18 prodrugs increase the frequency of CD8+ T cells and decreases the frequency of M2 macrophages at both timepoints. The effect was greater with WW50480 / 50487.

[0068] FIGs.25A and 25C show the frequency of MC38 tumor infiltrating Natural Killer (NK) cells that were producing IFNγ or Granzyme B on the either Day 5 (FIG.25A) or Day 11 (FIG.25C). FIGs. 25B and 25D shows the frequency of MC38 tumor infiltrating Natural Killer (NK) cells that were producing either 0, 1, 2, or 3 effector cytokines simultaneously on either Day 5 (FIG.25B) or Day 11 (FIG.25D) after ex vivo restimulation for intracellular cytokine staining. The data show that treatment with WW50480 / 50487 increases the frequency of NK cells producing effector cytokines on Day 5. Similarly, treatment with inducible IL-18 prodrugs increased the frequency of NK cells producing effector cytokines on Day 11. The effect was greater with WW50480 / 50487. On Day 5, treatment with WW50480 / 50487 drove NK cells to produce more than one effector cytokine simultaneously (polyfunctionality).

[0069] FIGs.26A and 26C are graphs showing the frequency of MC38 tumor infiltrating Natural Killer T (NKT) cells that were producing IFNγ, Granzyme B on either Day 5 (FIG.26A) or Day 11 (FIG.26C). FIGs.26B and 26D shows the frequency of MC38 tumor infiltrating NKT cells that were producing either 0, 1, 2, or 3 effector cytokines simultaneously on either Day 5 (FIG.26B) or Day 11 (FIG.26D) after ex vivo restimulation for intracellular cytokine staining. The data show that treatment with WW50480 / 50487 increases the frequency of NKT cells producing effector cytokines on Day 5, as well as the frequency of NKT cells producing multiple effector cytokines (polyfunctionality). Similarly, treatment with inducible IL-18 prodrug molecules increases the frequency of NKT cells producing individual effector cytokines or multiple effector cytokines at once on Day 11, though the effect was greater with WW50480 / 50487.

[0070] FIGs.27A and 26C are graphs showing the frequency of MC38 tumor infiltrating tetramer positive CD8+ T cells that were producing IFNγ, TNF, or Granzyme B on either Day 5 (FIG.27A) or Day 11 (FIG.27C). FIGs.26B and 26D shows the frequency of MC38 tumor infiltrating tetramer positive CD8+ T cells that were producing either 0, 1, 2, 3, or 4 effector cytokines simultaneously on either Day 5 (FIG.27B) or Day 11 (FIG.27D) after ex vivo restimulation for intracellular cytokine staining. The data show that treatment with WW50480 / 50487 increases the frequency of tetramer positive 15 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 CD8+ T cells producing effector cytokines on Day 5, as well as the frequency of tetramer+ CD8+ T cells producing multiple effector cytokines (polyfunctionality). Similarly, treatment with inducible IL-18 prodrugs increases the frequency of tetramer positive CD8+ T cells producing individual effector cytokines or multiple effector cytokines at once on Day 11, though the effect was greater with WW50480 / 50487.

[0071] FIGs.28A and 28C are graphs showing the frequency of MC38 tumor infiltrating B cells, M1 macrophages, or M2 macrophages producing IFNγ on either Day 5 (FIG.27A) or Day 11 (FIG.27C). FIGs.28B and 28D are graphs showing the frequency of M1 and M2 macrophages (FIG.27B) or the frequency of M2 macrophages producing arginase and iNOS (FIG.27D). The data show that treatment with WW50480 / 50487 increases the frequency of B cells, M1 macrophages, and M2 macrophages producing IFNγ. Similarly, treatment with WW50480 / 50487 increases the frequency of M1 macrophages, and decreases the frequency M2 macrophages, while increasing the frequency of activated M2 macrophages producing arginase and iNOS.

[0072] FIGs.29A-29C are graphs showing the levels of IL-18 binding protein (BP) measured in the tumor (FIG.29A), spleen (FIG.29B), or plasma (FIG.29C) of tumor bearing animals after treatment on day 5 and day 11 with either WW50480 / WW50487 (WT prodrug) or WW50482 / WW50487 (BPR prodrug). The data show that treatment with WW50480 / 50487 increases the amount of IL-18BP found in the tumor on Day 5, and increases the amount of IL-18BP the spleen and plasma at both timepoints, consistent with ongoing IL-18 signaling in vivo. In contrast, increased IL-18BP was found in the spleen of animals treated with WW50482 / 50487 on Day 11, but not in the other location at the other tested timepoints, consistent with an IL-18BP enforced negative feedback loop inhibiting ongoing IL-18 signaling.

[0073] FIGs.30A-30I are graphs showing the levels of IFNγ measured in the tumor (FIG.30A) and plasma (FIG.30E) or the levels of other cytokines (TNFα (FIG.30B), IP-10 (FIG.30C), IL-6 (FIG.30D), IL-5 (FIG.30F), IL-4 (FIG.30G), IL-1β (FIG.30H), and IL-10 (FIG.30I) in the plasma after treatment on day 5 and day 11 with either WW50480 / WW50487 (WT prodrug) or WW50482 / WW50487 (BPR prodrug) . The data show that WW50480 / 50487 treatment increased the levels of IFNγ in the tumor on Day 5, with limited effects on the plasma levels of this cytokine. In contrast, treatment with WW50482 / 50487 increased the level of IFNγ detected in the plasma at both timepoints. Similar increases in plasma levels of TNFα, IL-5, IL-4, IL-1β, and IL-10 were observed at various timepoints selectively following WW50482 / 50487 treatment. Increased plasma levels of IL-6 were observed following WW50480 / 50487 treatment, while both inducible IL-18 prodrugs containing molecules increased plasma levels of IP-10. 16 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0074] FIG.31 is a graph showing the tumor level of IL-18 bound to IL-18BP (IL18:IL18BP) after administration of vehicle, BPR inducible mIL-18 prodrug, or WT inducible mIL-18 prodrug. Dosing with WT inducible mIL-18 prodrug resulted in a nearly 2-fold increased initial IL18:IL18BP exposure in the tumor by Day 5 and decayed to baseline by the end of the observation period. In contrast, tumor level of IL18:IL18BP was consistently low and barely detectable above the limit of quantification either on Day 5 or Day 12 in any of the animals treated with inducible BPR mIL-18 prodrug, indicating that BPR IL-18 is resistant to inhibition by IL-18 BP.

[0075] FIGs.32A-32D show inducible BPR mouse IL-18 prodrug drives more infiltration of activated polyfunctional CD8+ T-cells. FIG.32A shows the infiltration of polyfunctional (expressing both Granzyme B and Perforin) CD8+ T cells into the tumor after treatment with either vehicle, a WT IL-18 containing inducible IL-18 prodrug (WW50629 / WW50632), or an inducible BPR IL-18 prodrug (WW50941 / WW50632) using density heatmaps (HALO software). FIG.32B shows density plot representation of tumor cells, CD8+ polyfunctional T cells and stem-like CD8+ T cells and the spatial location of these cells within the tumor tissue. FIG.32C is a graph showing quantification of the overall density with 3 tumors per treatment. FIG.32D is a graph showing tumor penetration of polyfunctional (Granzyme B+ Perforin+) CD8+ T cells. (C) One-Way or D) Two-Way ANOVAs were performed, and significance is reported as follows: *p<0.05, **p<0.005 , **** p<0.0001,*** p<0.0005. The data demonstrate that only treatment with an inducible BPR IL-18 prodrug (WW50941 / WW50632) was able to drive polyfunctional CD8+ T cells deep into the tumor tissue, while treatment with an inducible WT IL-18 prodrug (WW50629 / WW50632) failed to generate any activity distinct from vehicle treatment.

[0076] FIGs.33A-33C show the infiltration of stem-like TCF1+ CD8+ T cells into the tumor after treatment with either vehicle, a WT inducible IL-18 prodrug (WW50629 / WW50632), or a BPR inducible IL-18 prodrug (WW50941 / WW50632). FIG.33A is a density heatmap of TCF1+ CD8+ T cells generated using HALO software (3 tumors per treatment) FIG.33B is a graph showing quantification of TCF1+ CD8+ T cell frequency. FIG.33C is a graph showing density within the tumor of TCF1+ CD8+ T cell frequency. One-Way ANOVAs were performed, and significance is reported as follows: *= p<0.05. The data demonstrate that only treatment with a BPR inducible IL-18 prodrug (WW50941 / WW50632) was able to induce dense pockets of TCF1+ CD8+ T cells in the tumor tissue, while treatment with a WT inducible IL-18 prodrug (WW50629 / WW50632) failed to generate any activity distinct from vehicle treatment.

[0077] FIGs.34A-34C show the interactions and relative distances of polyfunctional CD8+ T cells with tumor cells after treatment with either vehicle, an inducible WT IL-18 prodrug (WW50629 / WW50632), or an inducible BPR IL-18 prodrug (WW50941 / WW50632). FIG.34A shows representative multiplex immunofluorescence (M-IF) images of polyfunctional Granzyme B+ Perforin+ CD8+ T cells interacting 17 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 with SOX2+ tumor cells. FIG.34B is a graph showing the quantification of the frequency of tumor cells with an adjacent polyfunctional CD8+ T cell. FIG.34C is a graph showing average distances between tumor cells and polyfunctional CD8+ T cells. One-Way ANOVAs were performed, and significance is reported as follows: ** p<0.005. The data demonstrate that only treatment with an inducible BPR IL-18 prodrug (WW50941 / WW50632) was able to increase the frequency of tumor cells with directly adjacent (≤10µm) polyfunctional CD8+ T cells, while treatment with an inducible WT IL-18 prodrug (WW50629 / WW50632) failed to generate any activity distinct from vehicle treatment.

[0078] FIGs.35A and 35B show the activation of tumor infiltrating NK cells after treatment with either vehicle, an inducible WT IL-18 prodrug (WW50629 / WW50632), or an inducible BPR IL-18 prodrug (WW50941 / WW50632). FIG.35A shows representative M-IF images. FIG.35B is a graph showing quantification of activated NK cells. A One-Way ANOVA was performed, and significance is reported as follows: *p<0.05; ** p<0.005. The data demonstrate that treatment with either an inducible WT IL-18 prodrug (WW50629 / WW50632) or a inducible BPR IL-18 prodrug (WW50941 / WW50632) was sufficient to increase the frequency of NK cells producing both perforin and Granzyme B.

[0079] FIGs.36A and 36B are graphs showing the levels of IFNgamma (FIG.36A) and IL-18 binding protein (BP) (FIG.36B) measured tumor lysates on day 5, 24 hrs after the second dose in MC38 tumor bearing mice dosed with vehicle, inducible WT-IL-18 prodrug, or inducible IL-18BP prodrug. The data show that treatment with WW50629 / WW50632 or WW50941 / WW50632 increases the amount of IL- 18BP and IFNγ found in the tumor 24 hours after dosing.

[0080] FIGs.37A-37D show a series of graphs showing the activity of a inducible BPR IL-18 prodrug on tumor infiltrating immune cells. FIGs.37A and 37B shows the effects of BPR IL-18 on the frequency of tumor infiltrating NK cells producing at least three of the following four effector cytokines IFNγ, TNF, Perforin, Granzyme B (polyfunctionality). Statistics represent a two-way ANOVA where significance is reported as follows: *= p<0.05; ***= p<0.001. FIG.37c shows a representation of the molecular definitions of Naïve (TCF1+TOX-), Tpex (TCF1+TOX+), Tex-eff (TCF1-TOX-), and Tex (TCF1- TOX+). FIG.37d shows the average frequency of those populations within the CD8+ T cell population upon each treatment at 2 different time points. Results confirm that treatment of MC38 tumor bearing mice with an induclble BPR IL-18 prodrug has a unique and robust effect on NK cell activation in the tumor microenvironment. Furthermore, these results demonstrate that only treatment with a inducible BPR IL-18 prodrug was able to generate and maintain a population of CD8+ T cells with a stem-cell like T Progenitor of Exhaustion (Tpex) phenotype (TCF1+TOX+).

[0081] FIGs.38A-38D are graphs showing the activity of an inducible BPR IL-18 prodrug on several individual transcripts measured in bulk RNA from the dissociated tumors on Day 8. The individual transcripts measured are labeled on each graph, and include Ccl24, Batf, Slamf6, and Ccl17. Each of 18 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 these transcripts has been associated with the formation of immune hubs, indicating that inducible BPR IL-18 prodrug treatment uniquely induces a transcriptional environment conducive to immune hub formation. 4. DETAILED DESCRIPTION A. Inducible Cytokine Prodrugs

[0082] The disclosure relates to inducible IL-18 prodrugs that contain at least one polypeptide chain, and can contain two or more polypeptide chains, if desired. The inducible IL-18 prodrugs comprise an IL-18 or a variant thereof, a half-life extension element, an IL-18 blocking element, and a protease cleavable linker. The inducible IL-18 prodrugs can be encoded by the nucleic acids disclosed herein.

[0083] The inducible IL-18 prodrugs of this disclosure have attenuated IL-18 receptor agonist activity and the circulating half-life is extended. The IL-18 receptor agonist activity is attenuated through the blocking element, which is capable of blocking all or some of the receptor agonist activity of the IL-18 or variant within the prodrug, typically by noncovalently binding to the IL-18 or variant within the prodrug and / or sterically blocking their binding to the receptor. The half-life extension element can also contribute to attenuation, for example through steric effects. The half-life extension element can also act as a blocking element that is capable of blocking all or some of the receptor agonist activity of IL-18. For instance, the half-life extension element can contribute to blocking when the half-life extension element is adjacent to the IL-18 polypeptide.

[0084] Upon cleavage of the protease cleavable linker, a form of IL-18 is released that is active (e.g., more active than the inducible IL-18 prodrug). Typically, the released IL-18 is at least 10 x more active than the inducible IL-18 prodrug. Preferably, the released IL-18 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-18 prodrug. Typically, cleavage of the protease cleavable linker releases IL- 18 from both the IL-18 blocking element and the half-life extension element.

[0085] The form of IL-18 that is released upon cleavage of the inducible IL-18 prodrug typically has a short half-life, which is often substantially similar to the half-life of naturally occurring IL-18. Even though the half-life of the inducible IL-18 prodrug is extended, toxicity is reduced or eliminated because the agonist activity of the circulating inducible IL-18 prodrug is attenuated and active IL-18 is targeted to the desired site of activity (e.g., tumor microenvironment).

[0086] It will be appreciated by those skilled in the art, that the number of polypeptide chains, and the location of the elements, the half-life extension element, the protease cleavable linker(s), and the blocking element (and components of such elements, such as a VH or VL domain) on the polypeptide chains can vary and is often a matter of design preference. All such variations are encompassed by this disclosure. 19 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0087] The inducible IL-18 prodrug can comprise a single polypeptide chain. Typically, the single polypeptide chain comprises an IL-18 polypeptide [A], a half-life extension element [H], a blocking domain [D], and a protease cleavable linker [L]. The IL-18 polypeptide [A] can be operably linked to the half-extension element, the blocking domain, or both the half-life extension element and blocking domain by a protease cleavable linker. Typically, the single polypeptide chain comprises one IL-18 polypeptide or two IL-18 cytokine polypeptides. The IL-18 polypeptide can be located at any desired position in the single polypeptide chains.

[0088] The single polypeptide can comprise two or more half-life extension elements that also function as a blocking domain. When two or more half-life extension elements are present in the inducible IL-18 prodrug, they can block all or some of the receptor agonist activity of the IL-18 and also extend serum half-life. When two or more half-life extension elements are present in the inducible cytokine prodrug, a separate blocking domain is optional and is typically not present.

[0089] The inducible IL-18 prodrug can be of any of Formulas (I)-(VI): [A]-[L1]-[H]-[L2]-[D] (I); [D]-[L2]-[H]-[L1]-[A] (II); [A]-[L1]-[D]-[L2]-[H] (III); [H]-[L2]-[D]-[L1]-[A] (IV); [H]-[L1]-[A]-[L2’]-[D] (V); and [D]-[L1]-[A]-[L2’]-[H] (VI).

[0090] In Formulas (I) – (VI), [A] is an IL-18 polypeptide, [D] is a blocking domain, [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 [L2’] is a protease-cleavable polypeptide linker. [L1] and [L2] or [L1] and [L2’] can have the same or different amino acid sequence and or protease-cleavage site (when L2 is protease-cleavable) as desired. The protease cleavable linker can be a linker as disclosed herein and, for example, can comprise the sequence GPAGLYAQ (SEQ ID NO: 426) or ALFKSSFP (SEQ ID NO: 357), particularly ALFKSSFP (SEQ ID NO: 357). Preferably, each of [L1] and [L2’] comprises SEQ ID NO: 354 or 357.

[0091] While the inducible IL-18 prodrugs disclosed herein may contain one half-life extension element and one blocking element, such elements 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 (i.e., the inducible IL-18 prodrugs can contain two or more polypeptide chains). Such inducible IL-18 prodrugs comprise an IL-18 polypeptide, a half-life extension element, a blocking domain, and a protease cleavable linker. For example, a first polypeptide chain can include an antibody light chain (VL+CL) or 20 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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 peptide complex to form an antigen-binding site, such as a Fab that binds to the IL-18 and attenuates the IL-18 activity.

[0092] For example, the inducible IL-18 prodrug can have a) a first polypeptide of Formulas (I)-(VI), wherein [A] is a IL-18 polypeptide, [D] is an antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH), [H] is a half-life extension element, [L1] is a protease-cleavable polypeptide linker, [L2] is an polypeptide linker that is optionally protease-cleavable, and [L2’] is a protease-cleavable polypeptide linker; and b) a second polypeptide chain comprising an antibody light chain (VL+CL) or light chain variable domain (VL) that is complementary to the VH + CH1 or VH. [L1] and [L2] or [L1] and [L2’] can have the same or different amino acid sequence and or protease-cleavage site (when L2 is protease-cleavable) as desired.

[0093] For example, the inducible IL-18 prodrug can have a) a first polypeptide of Formulas (I)-(VI), wherein [A] is a IL-18 polypeptide, [D] is an antibody light chain (VL + CL) or light chain variable domain (VL), [H] is a half-life extension element, [L1] is a protease-cleavable polypeptide linker, [L2] is an polypeptide linker that is optionally protease-cleavable, and [L2’] is a protease-cleavable polypeptide linker; and b) a second polypeptide an antibody heavy chain Fab fragment (VH + CH1) or heavy chain variable domain (VH) that is complementary to the VL + CL or VL. [L1] and [L2] or [L1] and [L2’] can have the same or different amino acid sequence and or protease-cleavage site (when L2 is protease- cleavable) as desired.

[0094] For instance, the first polypeptide chain can comprise from the amino terminal to the carboxy terminal the IL-18 polypeptide, a protease cleavable linker, a half-life extension element, and a VH and CH1 of an antibody that binds the IL-18. The second polypeptide chain can comprise a VL and CL of an antibody that binds the IL-18 and that together with the VH and CH1 of the first polypeptide chain form a Fab that binds the IL-18 polypeptide.

[0095] The inducible IL-18 prodrugs disclosed herein can comprise two blocking elements on two different polypeptide chains. The first polypeptide chain can comprise IL-18Rα and the second polypeptide chain can comprise the IL-18Rβ.

[0096] In embodiments, at least one of the 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-18 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-18 through a protease cleavable linker. The second polypeptide can comprise the half-life extension element and either an antibody light chain (VL + CL) or 21 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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-18. In an example, the first polypeptide can comprise IL-18 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-18. In another example, the first polypeptide can comprise IL-18 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-18.

[0097] 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- 18 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-18 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-18 polypeptide.

[0098] In embodiments, each polypeptide chain can contain one IL-18 polypeptide, one half-life extension element, and one blocking element. For example, a first polypeptide chain can comprise an IL- 18 polypeptide, a half-life extension element, and a blocking element, and the second polypeptide chain can comprise an IL-18 polypeptide, a half-life extension element, and a blocking element i. IL-18 polypeptide

[0099] As disclosed herein, the inducible IL-18 prodrug includes an IL-18 polypeptide. Any suitable IL- 18 polypeptide can be used in the IL-18 prodrugs disclosed herein. The IL-18 polypeptide can comprise or consist of the amino acid sequence of any one of SEQ ID NOs: 2-170, 200-209, or 211-245, or an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 22 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 2-170, 200-209, or 211-245. The IL-18 polypeptide can comprise or consist of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 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, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 200, 201, 202, 203, 204, 204, 206, 207, 208, 209, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, or 245.

[0100] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 2, or an amino acid sequence that has at least about 95% identity, about 90%, preferably about 95%, about 96%, about 97%, about 98%, or about 99% to SEQ ID NO: 2. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 2, wherein the amino acid at position 56 is deleted.

[0101] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 3, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 3. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 3 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 3 are not substituted.

[0102] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 4, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 4. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 4 wherein the amino acid at position 10 of SEQ ID NO: 4 is not substituted.

[0103] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 5, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 5. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 5 wherein the amino acids at positions 10, 49 and 62 of SEQ ID NO: 5 are not substituted.

[0104] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 6, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 6. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% 23 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 identity to SEQ ID NO: 6 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO:6 are not substituted.

[0105] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 7. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 7 wherein the amino acid at position 10 of SEQ ID NO: 7 is not substituted.

[0106] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8 wherein the amino acids at positions 57 and 62 of SEQ ID NO: 8 are not substituted.

[0107] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 9. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 9 wherein the amino acids at positions 62 and 91 of SEQ ID NO: 9 are not substituted.

[0108] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 10. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 10 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 10 are not substituted.

[0109] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 11. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 11 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 11 are not substituted.

[0110] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 12. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 12 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 12are not substituted.

[0111] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 13, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 13. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 24 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 95% identity to SEQ ID NO: 13 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 13 are not substituted.

[0112] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 14, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 14 are not substituted.

[0113] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 15, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 15. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 15 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 15 are not substituted.

[0114] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 16, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 16. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 16 wherein the amino acid at position 10 of SEQ ID NO: 16 is not substituted.

[0115] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 17, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 17. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 17 wherein the amino acids at positions 10, 49, 62 and 91 of SEQ ID NO: 17 are not substituted.

[0116] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 18, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 18. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 18 wherein the amino acid at position 91 of SEQ ID NO: 18 is not substituted.

[0117] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 19, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 19. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 19 wherein the amino acid at position 91 of SEQ ID NO: 19 is not substituted.

[0118] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 20, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 20. For example, 25 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 20 wherein the amino acid at position 91 of SEQ ID NO: 20 is not substituted.

[0119] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 21, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 21. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 21 wherein the amino acid at positions 10 and 91 of SEQ ID NO: 21 is not substituted.

[0120] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 22, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 22. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 22 wherein the amino acids at position 10, 51, and 91 of SEQ ID NO: 22 is not substituted.

[0121] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 23, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 23. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 23 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 23 are not substituted.

[0122] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 24, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 24. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 24 wherein the amino acid at position 91 of SEQ ID NO: 24 is not substituted.

[0123] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 25, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 25 are not substituted.

[0124] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 26, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 26. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 26 wherein the amino acid at position 91 of SEQ ID NO: 26 is not substituted. 26 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0125] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 27, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27 wherein the amino acids at positions 49 and 57 of SEQ ID NO: 27 is not substituted.

[0126] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 28, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 28. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 28 wherein the amino acid at position 10 or 57 of SEQ ID NO: 28 is not substituted.

[0127] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 29, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 29. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 29 wherein the amino acid at position 10 of SEQ ID NO: 29 is not substituted.

[0128] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 30, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 30. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 30 wherein the amino acid at position 91 of SEQ ID NO: 30 is not substituted.

[0129] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 31, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 31. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 31 wherein the amino acid at position 49 and 57 of SEQ ID NO: 31 is not substituted.

[0130] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 32, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 32. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 32 wherein the amino acids at positions 60 and 91 of SEQ ID NO: 32 are not substituted.

[0131] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 33, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 33. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 27 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 95% identity to SEQ ID NO: 33 wherein the amino acids at positions 10, 49, 57, and 91 of SEQ ID NO: 33 are not substituted.

[0132] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 34, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 34. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 34 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 34 are not substituted.

[0133] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 35, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 35. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 35 wherein the amino acid at position 91 of SEQ ID NO: 35 is not substituted.

[0134] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 36, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 36. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 36 wherein the amino acid at position 91 of SEQ ID NO: 36 is not substituted.

[0135] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 37, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 37. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 37 wherein the amino acid at position 91 of SEQ ID NO: 37 is not substituted.

[0136] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 38, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 38. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 38 wherein the amino acids at position 10 of SEQ ID NO: 38 is not substituted.

[0137] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 39, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 39. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 39 wherein the amino acid at position 91 of SEQ ID NO: 39 is not substituted.

[0138] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 40, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 40. For example, 28 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 40 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 50 are not substituted.

[0139] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 41, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 41. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 41 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 41 are not substituted.

[0140] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 42, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 42. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 42 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 42 are not substituted.

[0141] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 43, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 43. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 43 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 43 are not substituted.

[0142] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 44, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 44. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 44 wherein the amino acids at positions 10 and 49 of SEQ ID NO: 44 are not substituted.

[0143] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 45, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 45. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 45 wherein the amino acid at position 60 and 91 of SEQ ID NO: 45 is not substituted.

[0144] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 46, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 46. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 46 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 46 are not substituted. 29 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0145] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 47, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 47. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 47 wherein the amino acids at positions 3 and 54 of SEQ ID NO: 47 are not substituted.

[0146] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 48, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 48. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 48 wherein the amino acid at position 54 of SEQ ID NO: 48 is not substituted.

[0147] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 49, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 49. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 49 wherein the amino acids at positions 3 and 54 of SEQ ID NO: 49 are not substituted.

[0148] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 50, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 50. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 50 wherein the amino acids at positions 3 and 6 of SEQ ID NO: 50 are not substituted.

[0149] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 51, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 51. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 51 wherein the amino acid at positions 3 and 6 of SEQ ID NO: 51 is not substituted.

[0150] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 52, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 52. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 52 wherein the amino acid at position 3 of SEQ ID NO: 52 is not substituted.

[0151] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 53, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 53. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 30 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 95% identity to SEQ ID NO: 53 wherein the amino acids at positions 3 and 6 of SEQ ID NO: 53 are not substituted.

[0152] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 54, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 54. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 54 wherein the amino acid at position 3 of SEQ ID NO: 54 is not substituted.

[0153] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 55, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 55. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 55 wherein the amino acids at positions 3 and 54 of SEQ ID NO: 55 are not substituted.

[0154] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 56. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 56 wherein the amino acids at positions 3 and 6 of SEQ ID NO: 56 are not substituted.

[0155] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 57, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 57. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 57 wherein the amino acid at position 3 of SEQ ID NO: 57 is not substituted.

[0156] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 58, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 58. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 58 wherein the amino acids at positions 36 and 37 of SEQ ID NO: 58 is not substituted.

[0157] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 59, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 59. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 59 wherein the amino acid at positions 36 and 40 of SEQ ID NO: 59 is not substituted.

[0158] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 60, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 60. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 31 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 95% identity to SEQ ID NO: 60 wherein the amino acids at positions 6, 36, and 40 of SEQ ID NO: 60 are not substituted.

[0159] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 61, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 61. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 61 wherein the amino acids at positions 36 and 37 of SEQ ID NO: 61 are not substituted.

[0160] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 62, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 62. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 62 wherein the amino acids at positions 36 and 37 of SEQ ID NO: 62 are not substituted.

[0161] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 63, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 63. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 63 wherein the amino acid at positions 36 of SEQ ID NO: 63 is not substituted.

[0162] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 64, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 64. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 64 wherein the amino acid at position 36 and 40 of SEQ ID NO: 64 is not substituted.

[0163] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 65, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 65. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 65 wherein the amino acids at positions 1, 51 and 53 of SEQ ID NO: 65 are not substituted.

[0164] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 66, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 66 are not substituted.

[0165] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 67, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 67. For example, 32 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 67 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 67 are not substituted.

[0166] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 68, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 68. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 68 wherein the amino acids at positions 56, 57 and 60 of SEQ ID NO: 68 are not substituted.

[0167] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 69, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 69. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 69 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 6 are not substituted.

[0168] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 70, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 70. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 70 wherein the amino acids at positions 36, 49, and 57 of SEQ ID NO: 70 are not substituted.

[0169] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 71, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71 wherein the amino acids at positions 36, 40, 49, and 57 of SEQ ID NO: 71 are not substituted.

[0170] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 72, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 72. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 72 wherein the amino acids at positions 36 and 54 of SEQ ID NO: 72 are not substituted.

[0171] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 73, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73 wherein the amino acids at positions 36, 40, and 54 of SEQ ID NO: 73 are not substituted.

[0172] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 74, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 74. For example, the IL-18 33 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 74 wherein the amino acids at positions 36, 51, and 53 of SEQ ID NO: 74 are not substituted.

[0173] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 75, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75 wherein the amino acids at positions 36, 40, 51, and 53 of SEQ ID NO: 75 are not substituted.

[0174] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 76, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 76. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 76 wherein the amino acids at positions 36, 51, and 53 of SEQ ID NO: 76 are not substituted.

[0175] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 77, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 77. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 77 wherein the amino acids at positions 36, 40, 51, and 53 of SEQ ID NO: 77 are not substituted.

[0176] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 78, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 78. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 78 wherein the amino acids at positions 10, 49, and 57 of SEQ ID NO: 78 are not substituted.

[0177] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 79, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 79. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 79 wherein the amino acids at positions 10, 36, 49, and 57 of SEQ ID NO: 79 are not substituted.

[0178] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 80, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 80. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 80 wherein the amino acids at positions 10, 36, 40, 49, and 57 of SEQ ID NO: 80 are not substituted. 34 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0179] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 81, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 81. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 80 wherein the amino acids at positions 1 is deleted.

[0180] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 82, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 82. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 82 wherein the amino acids at position 36 of SEQ ID NO: 82 is not substituted.

[0181] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 83, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83 wherein the amino acids at positions 36 and 40 of SEQ ID NO: 83 are not substituted.

[0182] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 84, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 84. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 84 wherein the amino acids at positions 51 and 57 of SEQ ID NO: 84 are not substituted.

[0183] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 85, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 85. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 85 wherein the amino acids at positions 36, 51 and 57 of SEQ ID NO: 85 are not substituted.

[0184] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 86, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 86. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 86 wherein the amino acids at positions 36, 40, 51 and 57 of SEQ ID NO: 86 are not substituted.

[0185] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 87, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 87. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 87 wherein the amino acids at positions 49 and 53 of SEQ ID NO: 87 are not substituted.

[0186] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 88, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 88. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 88 wherein the amino acids at positions 36, 49 and 53 of SEQ ID NO: 88 are not substituted. 35 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0187] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 89, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 89. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 89 wherein the amino acids at positions 36, 40, 49 and 53 of SEQ ID NO: 89 are not substituted.

[0188] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 90, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 90. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 90 wherein the amino acid at position 38 SEQ ID NO: 90 is not substituted.

[0189] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 91, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 91. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 91 wherein the amino acids at positions 38, 49 and 57 of SEQ ID NO: 91 are not substituted.

[0190] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 92, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 92. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 92 wherein the amino acids at positions 36, 38, 49 and 57 of SEQ ID NO: 92 are not substituted.

[0191] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 93, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 93. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 93 wherein the amino acids at positions 36, 38, 40, 49 and 57 of SEQ ID NO: 93 are not substituted.

[0192] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 94, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 94. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 94 wherein the amino acids at positions 38 and 54 of SEQ ID NO: 94 are not substituted.

[0193] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 95, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 95. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 95 wherein the amino acids at positions 36, 38 and 54 of SEQ ID NO: 95 are not substituted.

[0194] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 96, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 96. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 36 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 SEQ ID NO: 96 wherein the amino acids at positions 36, 38, 40 and 54 of SEQ ID NO: 96 are not substituted.

[0195] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 97, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 97. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 97 wherein the amino acids at positions 38, 51 and 53 of SEQ ID NO: 97 are not substituted.

[0196] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 98, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 98. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 98 wherein the amino acids at positions 36, 38, 51 and 53 of SEQ ID NO: 98 are not substituted.

[0197] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 99, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 99. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 99 wherein the amino acids at positions 36, 38, 40, 51 and 53 of SEQ ID NO: 99 are not substituted.

[0198] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 100, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 100. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 100 wherein the amino acids at positions 38, 51 and 53 of SEQ ID NO: 100 are not substituted.

[0199] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 101, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 101. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 101 wherein the amino acids at positions 36, 38, 51 and 53 of SEQ ID NO: 101 are not substituted.

[0200] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 102, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 102. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 102 wherein the amino acids at positions 36, 38, 40, 51 and 53 of SEQ ID NO: 102 are not substituted.

[0201] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 103, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 103. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 37 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 SEQ ID NO: 103 wherein the amino acids at positions 10, 49, 57, and 38 of SEQ ID NO: 103 are not substituted.

[0202] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 104, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 104. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 104 wherein the amino acids at positions 10, 36, 38, 49, and 57 of SEQ ID NO: 104 are not substituted.

[0203] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 105, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 105. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 105 wherein the amino acids at positions 10, 36, 38, 40, 49, and 57 of SEQ ID NO: 105 are not substituted.

[0204] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 106, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 106. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 106 wherein the amino acid at position 38 of SEQ ID NO: 106 is not substituted.

[0205] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 107, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 107. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 107 wherein the amino acids at positions 36 and 38 of SEQ ID NO: 107 are not substituted.

[0206] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 108, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 108. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 108 wherein the amino acids at positions 36, 38 and 40 of SEQ ID NO: 108 are not substituted.

[0207] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 109, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 109. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 109 wherein the amino acids at positions 36, 38 and 40 of SEQ ID NO: 109 are not substituted.

[0208] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 110, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 110. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 110 wherein the amino acids at positions 36 and 38 of SEQ ID NO: 110 are not substituted. 38 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0209] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 111, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 111. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 111 wherein the amino acids at positions 38, 51 and 57 of SEQ ID NO: 111 are not substituted.

[0210] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 112, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 112. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 112 wherein the amino acids at positions 36, 38, 51 and 57 of SEQ ID NO: 112 are not substituted.

[0211] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 113, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 113. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 113 wherein the amino acids at positions 36, 38, 40, 51 and 57 of SEQ ID NO: 113 are not substituted.

[0212] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 114, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 114. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 114 wherein the amino acids at positions 38, 49 and 53 of SEQ ID NO: 114 are not substituted.

[0213] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 115, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 115. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 115 wherein the amino acids at positions 36, 38, 49 and 53 of SEQ ID NO: 115 are not substituted.

[0214] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 116, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 116. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 116 wherein the amino acids at positions 36, 38, 40, 49 and 53 of SEQ ID NO: 116 are not substituted.

[0215] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 117, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 117. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 117 wherein the amino acid at position 185 of SEQ ID NO: 117 is not substituted. 39 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0216] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 118, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 118. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 118 wherein the amino acids at positions 49, 57, 162 and 185 of SEQ ID NO: 118 are not substituted.

[0217] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 119, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 119. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 119 wherein the amino acids at positions 36, 49, 57, 162 and 185 of SEQ ID NO: 119 are not substituted.

[0218] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 120, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120 wherein the amino acids at positions 36, 40, 49, 57, 162 and 185 of SEQ ID NO: 120 are not substituted.

[0219] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 121, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 121. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 121 wherein the amino acids at positions 54, 162 and 185 of SEQ ID NO: 121 are not substituted.

[0220] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 122, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 122. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 122 wherein the amino acids at positions 36, 54, 162 and 185 of SEQ ID NO: 122 are not substituted.

[0221] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 123, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 123. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 123 wherein the amino acids at positions 36, 40, 54, 162 and 185 of SEQ ID NO: 123 are not substituted.

[0222] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 124, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 124. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 40 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 SEQ ID NO: 124 wherein the amino acids at positions 51, 53, 162 and 185 of SEQ ID NO: 124 are not substituted.

[0223] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 125, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 125. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 125 wherein the amino acids at positions 36, 51, 53, 162 and 185 of SEQ ID NO: 125 are not substituted.

[0224] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 126, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 126. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 126 wherein the amino acids at positions 36, 40, 51, 53, 162 and 185 of SEQ ID NO: 126 are not substituted.

[0225] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 127, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 127. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 127 wherein the amino acids at positions 51, 53, 162 and 185 of SEQ ID NO: 127 are not substituted.

[0226] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 128, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 128. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 128 wherein the amino acids at positions 36, 162 and 185 of SEQ ID NO: 128 are not substituted.

[0227] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 129, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 129. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 129 wherein the amino acids at positions 36, 40, 51, 53, 162 and 185 of SEQ ID NO: 129 are not substituted.

[0228] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 130, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 130. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 130 wherein the amino acids at positions 10, 49, 57, 162, and 185 of SEQ ID NO: 130 are not substituted.

[0229] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 131, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 131. For example, the IL- 41 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 131 wherein the amino acids at positions 10, 36, 49, 57, 162, and 185 of SEQ ID NO: 131 are not substituted.

[0230] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 132, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 132. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 132 wherein the amino acids at positions 10, 36, 40, 49, 57162, and 185 of SEQ ID NO: 132 are not substituted.

[0231] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 133, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 133. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 133 wherein the amino acids at positions 162 and 185 of SEQ ID NO: 133 are not substituted.

[0232] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 134, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 134. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 134 wherein the amino acids at positions 36, 162 and 185 of SEQ ID NO: 134 are not substituted.

[0233] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 135, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 135. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 135 wherein the amino acids at positions 36, 40, 162 and 185 of SEQ ID NO: 135 are not substituted.

[0234] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 136, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 136. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 136 wherein the amino acids at positions 36, 40, 162 and 185 of SEQ ID NO: 136 are not substituted.

[0235] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 137, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 137. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 137 wherein the amino acids at positions 36, 162 and 185 of SEQ ID NO: 137 are not substituted. 42 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0236] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 138, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 138. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 138 wherein the amino acids at positions 51, 162 and 185 of SEQ ID NO: 138 are not substituted.

[0237] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 139, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 139. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 139 wherein the amino acids at positions 36, 51, 57, 162 and 185 of SEQ ID NO: 139 are not substituted.

[0238] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 140, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 140. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 140 wherein the amino acids at positions 36, 51, 57, 40, 162 and 185 of SEQ ID NO: 140 are not substituted.

[0239] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 141, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 141. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 141 wherein the amino acids at positions 49, 53, 162 and 185 of SEQ ID NO: 141 are not substituted.

[0240] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 142, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 142. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 142 wherein the amino acids at positions 36, 49, 53, 162 and 185 of SEQ ID NO: 142 are not substituted.

[0241] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 143, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 143. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 143 wherein the amino acids at positions 36, 40, 49, 53, 162 and 185 of SEQ ID NO: 143 are not substituted.

[0242] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 144, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 144. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 43 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 SEQ ID NO: 144 wherein the amino acids at positions 162 and 185 of SEQ ID NO: 144 are not substituted.

[0243] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 145, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 145. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 145 wherein the amino acids at positions 49, 38, 57, 162 and 185 of SEQ ID NO: 145 are not substituted.

[0244] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 146, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 146. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 146 wherein the amino acids at positions 36, 38, 49, 57, 162 and 185 of SEQ ID NO: 146 are not substituted.

[0245] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 147, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 147. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 147 wherein the amino acids at positions 36, 38, 40, 49, 57, 162 and 185 of SEQ ID NO: 147 are not substituted.

[0246] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 148, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 148. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 148 wherein the amino acids at positions 38, 54, 162 and 185 of SEQ ID NO: 148 are not substituted.

[0247] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 149, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 149. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 149 wherein the amino acids at positions 36, 38, 54, 162 and 185 of SEQ ID NO: 149 are not substituted.

[0248] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 150, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150 wherein the amino acids at positions 36, 38, 40, 54, 162 and 185 of SEQ ID NO: 150 are not substituted.

[0249] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 151, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 151. For example, the IL- 44 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 151 wherein the amino acids at positions 38, 51, 53, 162 and 185 of SEQ ID NO: 151 are not substituted.

[0250] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 152, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 152. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 152 wherein the amino acids at positions 36, 38, 51, 53, 162 and 185 of SEQ ID NO: 152 are not substituted.

[0251] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 153, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 153. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 153 wherein the amino acids at positions 36, 38, 40, 51, 53, 162 and 185 of SEQ ID NO: 153 are not substituted.

[0252] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 154, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 154. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 154 wherein the amino acids at positions 38, 51, 53, 162 and 185 of SEQ ID NO: 154 are not substituted.

[0253] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 155, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 155. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 155 wherein the amino acids at positions 36, 38, 51, 53, 162 and 185 of SEQ ID NO: 155 are not substituted.

[0254] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 156, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 156. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 156 wherein the amino acids at positions 36, 38, 40, 51, 53, 162 and 185 of SEQ ID NO: 156 are not substituted.

[0255] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 157, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 157. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 157 wherein the amino acids at positions 10, 38, 49, 57, 162 and 185 of SEQ ID NO: 157 are not substituted. 45 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0256] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 158, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 158. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 158 wherein the amino acids at positions 10, 36, 38, 49, 57, 162 and 185 of SEQ ID NO: 158 are not substituted.

[0257] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 159, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 159. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 159 wherein the amino acids at positions 10, 36, 38, 40, 49, 57, 162 and 185 of SEQ ID NO: 159 are not substituted.

[0258] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 160, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 160. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 160 wherein the amino acids at positions 38, 162 and 185 of SEQ ID NO: 160 are not substituted.

[0259] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 161, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 161. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 161 wherein the amino acids at positions 36, 38, 162 and 185 of SEQ ID NO: 161 are not substituted.

[0260] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 162, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 162. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 162 wherein the amino acids at positions 36, 38, 40, 162 and 185 of SEQ ID NO: 162 are not substituted.

[0261] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 163, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 163. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 163 wherein the amino acids at positions 36, 38, 40, 162 and 185 of SEQ ID NO: 163 are not substituted.

[0262] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 164, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 164. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 46 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 SEQ ID NO: 164 wherein the amino acids at positions 36, 38, 162 and 185 of SEQ ID NO: 164 are not substituted.

[0263] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 165, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 165. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 165 wherein the amino acids at positions 38, 51, 57, 162 and 185 of SEQ ID NO: 165 are not substituted.

[0264] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 166, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 166. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 166 wherein the amino acids at positions 36, 38, 51, 57, 162 and 185 of SEQ ID NO: 166 are not substituted.

[0265] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 167, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 167. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 167 wherein the amino acids at positions 36, 38, 40, 51, 57, 162 and 185 of SEQ ID NO: 167 are not substituted.

[0266] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 168, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 168. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 168 wherein the amino acids at positions 38, 49, 53, 162, and 185 of SEQ ID NO: 168 are not substituted.

[0267] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 169, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 169. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 169 wherein the amino acids at positions 36, 38, 49, 53, 162, and 185 of SEQ ID NO: 169 are not substituted.

[0268] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 170, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 170. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 170 wherein the amino acids at positions 36, 38, 40, 49, 53, 162, and 185 of SEQ ID NO: 170 are not substituted.

[0269] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 200, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 200. 47 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0270] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 201, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 201.

[0271] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 202, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 202.

[0272] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 203, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 203.

[0273] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 204, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 204.

[0274] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 205, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 205.

[0275] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 206, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 206.

[0276] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 207, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 207.

[0277] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 208, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 208.

[0278] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 209, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 209.

[0279] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 211, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 211. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 211 wherein the amino acids at positions 51, 53, and 60 of SEQ ID NO: 211 are not substituted.

[0280] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 212, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 212. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 212 wherein the amino acids at positions 36, 37, 56, 57, and 60 of SEQ ID NO: 212 are not substituted.

[0281] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 213, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 213. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 213 wherein the amino acids at positions 36, 37, 56, 60, and 133 of SEQ ID NO: 213 are not substituted. 48 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0282] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 214, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 214. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 214 wherein the amino acids at positions 36, 37, 56, and 60 of SEQ ID NO: 214 are not substituted.

[0283] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 215, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 215. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 215 wherein the amino acids at positions 36, 37, 56, 60, and 133 of SEQ ID NO: 215 are not substituted.

[0284] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 216, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 216. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 216 wherein the amino acids at positions 36, 37, 56, 60, and 133 of SEQ ID NO: 216 are not substituted.

[0285] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 217, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 217. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 217 wherein the amino acids at positions 36, 37, 56, and 60 of SEQ ID NO: 217 are not substituted.

[0286] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 218, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 218. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 218 wherein the amino acids at positions 36, 37, 56, and 60 of SEQ ID NO: 218 are not substituted.

[0287] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 219, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 219. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 219 wherein the amino acids at positions 36, 37, 57, and 60 of SEQ ID NO: 219 are not substituted.

[0288] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 220, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 220. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least 49 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 about 95% identity to SEQ ID NO: 220 wherein the amino acids at positions 36, 37, and 60 of SEQ ID NO: 220 are not substituted.

[0289] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 221, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 221. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 221 wherein the amino acids at positions 36, 37, 60, and 133 of SEQ ID NO: 221 are not substituted.

[0290] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 222, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 222. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 222 wherein the amino acids at positions 36, 37, 60, and 133 of SEQ ID NO: 222 are not substituted.

[0291] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 223, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 223. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 223 wherein the amino acids at positions 36, 37, 57, and 60 of SEQ ID NO: 223 are not substituted.

[0292] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 224, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 224. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 224 wherein the amino acids at positions 36, 37, 59, and 60 of SEQ ID NO: 224 are not substituted.

[0293] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 225, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 225. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 225 wherein the amino acids at positions 36, 37, 57, and 60 of SEQ ID NO: 225 are not substituted.

[0294] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 226, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 226. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 226 wherein the amino acids at positions 36, 37, and 60 of SEQ ID NO: 226 are not substituted.

[0295] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 227, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 227. For 50 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 227 wherein the amino acids at positions 36, 37, 57, 59, and 60 of SEQ ID NO: 227 are not substituted.

[0296] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 228, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 228. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 228 wherein the amino acids at positions 36, 37, 57, 59, and 60 of SEQ ID NO: 228 are not substituted.

[0297] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 229, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 229. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 229 wherein the amino acids at positions 36, 51, and 53 of SEQ ID NO: 229 are not substituted.

[0298] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 230, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 230. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 230 wherein the amino acids at positions 51, 53, 129, 131, and 134 of SEQ ID NO: 230 are not substituted.

[0299] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 231, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 231. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 231 wherein the amino acids at positions 36, 37, 51, and 53 of SEQ ID NO: 231 are not substituted.

[0300] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 232, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 232. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 232 wherein the amino acids at positions 51, 53, and 145 of SEQ ID NO: 232 are not substituted.

[0301] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 233, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 233. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 233 wherein the amino acids at positions 51, 53, and 150 of SEQ ID NO: 233 are not substituted. 51 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0302] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 234, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 234. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 234 wherein the amino acids at positions 51, 53, 131, and 132 of SEQ ID NO: 234 are not substituted.

[0303] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 235, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 235. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 235 wherein the amino acids at positions 51, 53, and 149 of SEQ ID NO: 235 are not substituted.

[0304] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 236, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 236. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 236 wherein the amino acids at positions 36, 37, 49, and 57 of SEQ ID NO: 236 are not substituted.

[0305] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 237, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 237. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 237 wherein the amino acids at positions 36, 49, and 57 of SEQ ID NO: 237 are not substituted.

[0306] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 238, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 238. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 238 wherein the amino acids at positions 37, 49, 57, 93, and 134 of SEQ ID NO: 238 are not substituted.

[0307] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 239, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 239. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 239 wherein the amino acids at positions 24, 27, 57, 49, and 111 of SEQ ID NO: 239 are not substituted.

[0308] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 240, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 240. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least 52 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 about 95% identity to SEQ ID NO: 240 wherein the amino acids at positions 49 and 57 of SEQ ID NO: 240 are not substituted.

[0309] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 241, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 241. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 241 wherein the amino acids at positions 34, 49, and 57 of SEQ ID NO: 241 are not substituted.

[0310] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 242, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 242. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 242 wherein the amino acids at positions 49, 57, and 129 of SEQ ID NO: 242 are not substituted.

[0311] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 243, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 243. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 243 wherein the amino acids at positions 49, 57, 129, 131, and 134 of SEQ ID NO: 243 are not substituted.

[0312] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 244, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 244. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 244 wherein the amino acids at positions 36, 37, and 54 of SEQ ID NO: 244 are not substituted.

[0313] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 245, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 245. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 245 wherein the amino acids at positions 36, 37, 49, and 57 of SEQ ID NO: 245 are not substituted.

[0314] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8 wherein the amino acids at positions 57 and 62 of SEQ ID NO: 8 are not substituted.

[0315] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 14, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14. For 53 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 14 are not substituted.

[0316] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 18, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 18. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 18 wherein the amino acid at position 91 of SEQ ID NO: 18 is not substituted.

[0317] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 25, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 25 are not substituted.

[0318] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 27, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27 wherein the amino acids at positions 49 and 57 of SEQ ID NO: 27 is not substituted.

[0319] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 66, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 66 are not substituted.

[0320] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 71, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71 wherein the amino acids at positions 36, 40, 49, and 57 of SEQ ID NO: 71 are not substituted.

[0321] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 75, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75 wherein the amino acids at positions 36, 40, 51, and 53 of SEQ ID NO: 75 are not substituted. 54 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0322] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 73, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73 wherein the amino acids at positions 36, 40, and 54 of SEQ ID NO: 73 are not substituted.

[0323] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 83, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83 wherein the amino acids at positions 36 and 40 of SEQ ID NO: 83 are not substituted.

[0324] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 96, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 96. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 96 wherein the amino acids at positions 36, 38, 40 and 54 of SEQ ID NO: 96 are not substituted.

[0325] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 103, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 103. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 103 wherein the amino acids at positions 10, 49, 57, and 38 of SEQ ID NO: 103 are not substituted

[0326] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 120, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120 wherein the amino acids at positions 36, 40, 49, 57, 162 and 185 of SEQ ID NO: 120 are not substituted.

[0327] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 150, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150 wherein the amino acids at positions 36, 38, 40, 54, 162 and 185 of SEQ ID NO: 150 are not substituted. ii. Half-life Extension Element

[0328] As disclosed herein, the inducible IL-18 prodrugs include a half-life extension element. The half- life extension element increases the in vivo half-life and provides altered pharmacodynamics and 55 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 pharmacokinetics of the inducible IL-18 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-18 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.

[0329] 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.

[0330] 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 when adjacent to the IL-18 polypeptide.

[0331] 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 56 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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 sdAb), e.g., an antibody or fragment that binds serum albumin (particularly HSA), an immunoglobulin Fc or fragment thereof, or serum albumin (particularly HSA).

[0332] Preferably, the half-life extension element is a single domain antibody fragment (sdAb). The sdAb can comprise or consist of SEQ ID NO: 602-606, 608-654, or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 602 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 603 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 604 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 605 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 606 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 608 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 609 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 610 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 611 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 612 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 613 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 614 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 615 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 616 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 617 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 618 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 619 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 620 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 621 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 622 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 623 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 624 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 625 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 625 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 626 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 627 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 628 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 629 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 630 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 631 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 632 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 633 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 634 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 635 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 636 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 637 or a variant thereof. 57 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 The sdAb can comprise or consist of SEQ ID NO: 638 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 639 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 640 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 641 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 642 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 643 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 644 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 645 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 46 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 647 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 648 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 647 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 649 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 647 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 650 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 647 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 651 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 647 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 652 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 647 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 653 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 647 or a variant thereof. The sdAb can comprise or consist of SEQ ID NO: 654 or a variant thereof.

[0333] The sdAb can comprise or consist of the amino acid sequence of any of SEQ ID NOs: 602-606, 608-654, or an amino acid sequence that has at least about 80% identity to any of SEQ ID NOs: 608-654. For example, the sdAbs can comprise or consist of an amino acid sequence that has at least about 81% identity, at least about 82% identity, at least about 83% identity, at least about 84% identity, at least about 85% identity, at least about 86% identity, at least about 87% identity, at least about 88% identity, at least about 89% identity, at least about 90% identity, at least about 91% identity, at least about 92% identity, at least about 93% identity, at least about 94% identity, at least about 95% identity, at least about 96% identity, at least about 97% identity, at least about 98% identity, or at least about 99% identity to any of SEQ ID NOs: 602, 603, 604, 605, 606, 608, 609, 610, 611, 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622, 623, 624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650, 651, 652, 653, or 654..

[0334] 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- 58 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 18 prodrugs disclosed herein can comprise a first half-life extension element and a second half-life 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.

[0335] 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).

[0336] In embodiments, the IL-18 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.

[0337] 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. 59 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0338] The half-life extension element of an inducible IL-18 prodrug extends the half-life of the inducible IL-18 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. iii. Blocking Element

[0339] The inducible IL-18 prodrugs disclosed herein comprise a blocking domain. The blocking domain can be any element that binds to the IL-18 and inhibits the ability of the IL-18 polypeptide to bind and activate its receptor. The blocking domain can inhibit the ability of the IL-18 to bind and / or activate its receptor e.g., by sterically blocking and / or by noncovalently binding to the IL-18 polypeptide. The blocking domain disclosed herein can bind to IL-18.

[0001] Examples of suitable blocking domains include the full length extracellular domain of IL-18 Receptor and an IL-18-binding fragment or variant thereof. The cognate receptor for IL-18 can be IL- 18Rα (IL18R1) and / or IL-18Rβ (IL18RAP). Antibodies and antigen-binding fragments thereof including, 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 sdAb and the like that bind to IL-18 can also be used. Other suitable antigen-binding domain that bind to the IL-18 polypeptide can also be used, 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, lipocallin and CTLA4 scaffolds.

[0340] Further examples of suitable blocking polypeptides include polypeptides that sterically inhibit or block binding of IL-18 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 IL-18 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 variant of such polypeptides. Blocking domains that are particularly suitable are single chain variable fragments (scFv) or Fab fragments.

[0341] IL-18 blocking elements that are particularly suitable are single chain variable fragments (scFv), Fab fragments, or full length or an IL-18-binding fragment or variant of the cognate receptor of an IL-18 (e.g., IL-18Rα receptor or the IL-18Rβ). Typically, the IL-18 blocking element will be an IL-18-binding Fab, dAb, scFv, or cognate IL-18 receptor or fragment thereof. 60 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0342] The blocking element can contain two or more components that are present on the same polypeptide chain or on separate polypeptide chains. 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 peptide complex to form an antigen-binding site, such as a Fab that binds IL-18 and attenuates IL-18 activity.

[0343] A first polypeptide chain can include a full length heavy chain (e.g., hIgG) and a second polypeptide can include an antibody light chain (VL+CL) or light chain variable domain (VL) that is complementary to the full length heavy chain (e.g., hIgG) on the first polypeptide. In such situations, these components can associate in the peptide complex to form an antigen-binding site that binds IL-18 and attenuates IL-18 activity.

[0344] The sdAbs disclosed herein can also act as blocking domains that are capable of blocking all or some of the receptor agonist activity of IL-18. For instance, the sdAb can contribute to blocking when the sdAb is adjacent to the IL-18 polypeptide.

[0345] Certain preferred IL-18 blocking elements are single chain variable fragments (scFv). The scFv can comprise or consist of the amino acid sequence of any one of SEQ ID NOs: 342, 343, 346, 347, 956, 957, 962, or 963 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 342, 343, 346, 347, 956, 957, 962, or 963.

[0346] The blocking element can comprise 1) a first polypeptide chain that comprises a heavy chain variable domain that can comprise or consist of an amino acid sequence of any one of SEQ ID NOs: 1033-1057 and 2) a second polypeptide chain that comprises or consists of a variable light chain that can comprise or consist of an amino acid sequence of any one of SEQ ID NOs: 1058-1082. The light chain variable is complementary to the heavy chain variable domain on the first polypeptide. These components can associate in the peptide complex to form an antigen-binding site that binds to IL-18 and attenuates IL- 18 activity. If desired, the heavy chain variable domain on the first polypeptide can comprise a heavy chain constant region that comprises or consists of SEQ ID NO: 1083. When present, the heavy chain variable domain and heavy chain constant domain form an antibody heavy chain Fab fragment (VH + CH1) on the first polypeptide chain. If desired, the light chain variable domain on the second polypeptide can comprise a light chain constant region that comprises or consists of SEQ ID NO: 1984. When present, the light chain variable domain and light chain constant domain form an antibody light chain Fab fragment on the second polypeptide chain(VL + CL). In such situations, these components (e.g., VH + CH1 and VL + CL) can associate in the peptide complex to form an antigen-binding site, such as a Fab that binds IL-18 and attenuates IL-18 activity. 61 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0347] Blocking elements 1-25 are specific examples of blocking elements comprising a heavy chain variable domain and a light chain variable domain. Further details of exemplary IL-18 antibody blocking elements are described in Table 1. Blocking Element Heavy chain variable domain Light chain variable domain Blocking Element 1 SEQ ID NO: 1033 SEQ ID NO: 1058

[0348] The blocking element can comprise 1) a first 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 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 172, or an amino acid sequence that has at least 95% identity SEQ ID NO: 172. 62 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0349] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide 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 and 2) a second polypeptide that comprises or consists 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.

[0350] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 318, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 318 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 319, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 319.

[0351] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 320, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 320 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 321, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 321.

[0352] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 322, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 322 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 323, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 323.

[0353] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 331, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 331 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 312, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 312.

[0354] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 353, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 353 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 864, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 864.

[0355] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 354, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 354 and 2) a second polypeptide that comprises or consists 63 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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.

[0356] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 655, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 655 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 865, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 865.

[0357] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 656, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 656 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 866, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 866.

[0358] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 657, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 657 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 314, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 314.

[0359] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 658, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 658 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 867, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 867.

[0360] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 734, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 734 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 869, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 869.

[0361] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 736, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 736 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 870, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 870.

[0362] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 738, or an amino acid sequence 64 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 that has at least 95% identity to SEQ ID NO: 738 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 871, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 871.

[0363] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 740, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 740 and 2) a second polypeptide that comprises or consists 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.

[0364] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 742, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 742 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 872, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 872.

[0365] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 875, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 875 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1012, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1012.

[0366] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 876, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 876 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1013, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1013.

[0367] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 877, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 877 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1014, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1014.

[0368] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 920, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 920 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1015, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1015. 65 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0369] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 329, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 329 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 311, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 311.

[0370] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 330, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 330 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 312, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 312.

[0371] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 337, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 337 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 862, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 862.

[0372] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 339, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 339 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 863, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 363.

[0373] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 659, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 659 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 864, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 864.

[0374] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 660, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 660 and 2) a second polypeptide that comprises or consists 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.

[0375] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 661, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 661 and 2) a second polypeptide that comprises or consists 66 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 the amino acid sequence of SEQ ID NO: 865, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 865.

[0376] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 662, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 662 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 866, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 866.

[0377] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 663, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 663 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 314, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 314.

[0378] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 664, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 664 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 867, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 867.

[0379] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 733, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 733 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 869, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 869.

[0380] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 735, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 735 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 870, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 870.

[0381] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 737, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 737 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 871, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 871.

[0382] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 739, or an amino acid sequence 67 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 that has at least 95% identity to SEQ ID NO: 739 and 2) a second polypeptide that comprises or consists 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.

[0383] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 741, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 741 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 872, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 872.

[0384] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 822, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 822 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 815, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 815.

[0385] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 823, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 823 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 816, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 816.

[0386] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 966, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 966 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1013, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1013.

[0387] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 967, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 967 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1014, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1014.

[0388] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 968, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 968 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1015, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1015. 68 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0389] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 969, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 969 and 2) a second polypeptide that comprises or consists the amino acid sequence of SEQ ID NO: 1016, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1016.

[0390] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1085, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1085 and 2) a second polypeptide 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 the amino acid sequence of SEQ ID NO: 187.

[0391] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1086, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1086, and 2) a second polypeptide 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 the amino acid sequence of SEQ ID NO: 187.

[0392] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1087, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1087, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 315.

[0393] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1088, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1088, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 315.

[0394] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1089, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1089, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1100, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 1100.

[0395] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1090, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1090, and 2) a second polypeptide that comprises or consists 69 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 of the amino acid sequence of SEQ ID NO: 1095, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 1095.

[0396] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1091, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1091, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1096, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 1096.

[0397] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1092, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1092, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1097, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 1097.

[0398] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1093, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1093, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1098, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 1098.

[0399] The blocking element can comprise 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1094, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 1094, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1099, or an amino acid sequence that has at least 95% identity to the amino acid sequence of SEQ ID NO: 1099. iii. Protease Cleavable Linker

[0400] As disclosed herein, the inducible IL-18 prodrug comprises one or more linker sequences. A linker sequence serves to provide flexibility between the polypeptides, such that, for example, the blocking domain is capable of inhibiting the activity of the IL-18. The linker can be located between the IL-18 subunit, the sdAb, and / or the blocking domain. As described herein the inducible IL-18 prodrug comprises a protease cleavable linker. The protease cleavable linker can comprise one or more cleavage sites for one or more desired protease. Preferably, the desired protease is enriched or selectively expressed at the desired target site of the IL-18 activity (e.g., the tumor microenvironment). Thus, the inducible IL- 18 prodrug is preferentially or selectively cleaved at the target site of desired IL-18 activity.

[0401] 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, 70 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, about 55, about 56, about 57, about 58, about 59, about 60, about 61, about 62, about 63, about 64, about 65, about 66, about 67, about 68, about 69, about 70, about 71, about 72, about 73, about 74, about 75, about 76, about 77, about 78, about 79, about 80, about 81, about 83, about 83, about 84, about 85, about 86, about 87, about 88, about 89, about 90, about 91, about 92, about 93, about 94, about 95, about 96, about 97, about 98, about 99m or about 100 amino acids in length. Preferred linkers are typically from about 5 amino acids to about 30 amino acids.

[0402] Preferably the lengths of linkers vary from 2 to 30 amino acids, optimized for each condition so that the linker does not impose any constraints on the conformation or interactions of the linked amino acid sequence of interest. 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 where 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 tom 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.

[0403] 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 tumor microenvironment, relative to the peripheral circulation.

[0404] 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 K, Cathepsin L, kallikreins, hKl, hK10, hK15, plasmin, collagenase, Type IV collagenase, stromelysin, Factor Xa, chymotrypsin-like protease, trypsin- like protease, 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, 71 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 matriptase, legumain, plasmepsin, nepenthesin, metalloexopeptidases, metalloendopeptidases, matrix metalloproteases (MMP), MMP1, MMP2, MMP3, MMP8, MMP9, MMP13, MMP11, MMP14, urokinase plasminogen activator (uPA), enterokinase, 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, a tumor cell surface protease, and an elastase. The MMP can, for example, be matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), matrix metalloproteinase 14 (MMP14). In addition, or alternatively, the linker can be cleaved by a cathepsin, such as, Cathepsin B, Cathepsin C, Cathepsin D, Cathepsin E, Cathepsin G, Cathepsin K and / or Cathepsin L. Preferably, the linker can be cleaved by MMP14 or Cathepsin L.

[0405] Proteases useful for cleavage of linkers and for use in the inducible IL-18 prodrugs herein are presented in Table 2, and exemplary proteases and their cleavage site are presented in Table 3. Table 2. Proteases relevant to inflammation and cancer Protease Specificity Other aspects Secreted by killer T cells: nt l h e; 72 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Protease Specificity Other aspects Associated with inflammation: Thr mbin Tr t: FGF2 T f rin r t m d lt ; e e n n Table 3. Exemplary Proteases and Protease Recognition Sequences Protease Cleavage Domain Sequence SEQ ID NO: 73 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Protease Cleavage Domain Sequence SEQ ID NO: MMP14 SGRIGFLRTA 385 74 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Protease Cleavage Domain Sequence SEQ ID NO: Kallikrein 2 GKAFRR 417 [ rs, 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 and / or a cathepsin.

[0407] Exemplary linkers that may be suitable for the inducible IL-18 prodrugs disclosed herein are disclosed in International Publication No.: WO2020 / 232305. For example, the linker can comprise the sequence GPAGLYAQ (SEQ ID NO: 426); GPAGMKGL (SEQ ID NO: 355); PGGPAGIG (SEQ ID NO: 356); ALFKSSFP (SEQ ID NO: 357); ALFFSSPP (SEQ ID NO: 358); LAQRLRSS (SEQ ID NO: 359); LAQKLKSS (SEQ ID NO: 360); GALFKSSFPSGGGPAGLYAQGGSGKGGSGK (SEQ ID NO: 361); RGSGGGPAGLYAQGSGGGPAGLYAQGGSGK (SEQ ID NO: 362); KGGGPAGLYAQGPAGLYAQGPAGLYAQGSR (SEQ ID NO: 363); RGGPAGLYAQGGPAGLYAQGGGPAGLYAQK (SEQ ID NO: 364); KGGALFKSSFPGGPAGIGPLAQKLKSSGGS (SEQ ID NO: 365); SGGPGGPAGIGALFKSSFPLAQKLKSSGGG (SEQ ID NO: 366); RGPLAQKLKSSALFKSSFPGGPAGIGGGGK (SEQ ID NO: 367); GGGALFKSSFPLAQKLKSSPGGPAGIGGGR (SEQ ID NO: 368); RGPGGPAGIGPLAQKLKSSALFKSSFPGGG (SEQ ID NO: 369); RGGPLAQKLKSSPGGPAGIGALFKSSFPGK (SEQ ID NO: 370); RSGGPAGLYAQALFKSSFPLAQKLKSSGGG (SEQ ID NO: 371); 75 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 GGPLAQKLKSSALFKSSFPGPAGLYAQGGR (SEQ ID NO: 372); GGALFKSSFPGPAGLYAQPLAQKLKSSGGK (SEQ ID NO: 373); RGGALFKSSFPLAQKLKSSGPAGLYAQGGK (SEQ ID NO: 374); RGGGPAGLYAQPLAQKLKSSALFKSSFPGG (SEQ ID NO: 375); SGPLAQKLKSSGPAGLYAQALFKSSFPGSK (SEQ ID NO: 376); KGGPGGPAGIGPLAQRLRSSALFKSSFPGR (SEQ ID NO: 377); KSGPGGPAGIGALFFSSPPLAQKLKSSGGR (SEQ ID NO: 378); or SGGFPRSGGSFNPRTFGSKRKRRGSRGGGG (SEQ ID NO: 379)

[0408] Certain preferred inducible IL-18 prodrugs comprise the sequence GPAGLYAQ (SEQ ID NO: 426) or ALFKSSFP (SEQ ID NO: 357).

[0409] Preferred linkers comprising more than one cleavage motif comprise the amino acids selected from SEQ ID NOs: 354 to 379. 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: 354 to 379.

[0410] The linker can comprise one or more cleavage motif or functional variants that are the same or different. The linker can comprise 1, 2, 3, 4, 5, or more cleavage motifs or functional variants. Linker 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: 354 to 379.

[0411] The length, degree of flexibility and other properties of the linkers used in the inducible IL-18 prodrugs may have some influence on properties, including, but not limited to the affinity, the specificity, or avidity for one or more components. In some instances, the linker can comprise flexible residues (such as glycine or serine) so that the adjacent sdAbs or amino acid sequence of interest are free to move relative to each other.

[0412] Other linker considerations include, but are not limited to, the effect on physical or pharmacokinetic properties of the resulting compound, such as solubility, lipophilicity, hydrophilicity, hydrophobicity, stability, rigidity, flexibility, immunogenicity, modulation of antibody binding, the ability to be incorporated into a micelle or liposome.

[0413] In some instances, a non-peptide linker may be suitable. For example, such linkage can be through covalent binding, affinity binding, intercalation, coordinate binding , and complexation. Covalent linking can be achieved by direct condensation of existing side chains or by the incorporation of external 76 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 bridging molecules. Linking agents can be useful for linking the sdAb thereof and the amino acid sequence together. For example, representative coupling agents include organic compounds such as thioesters, carbodiimide, succinimide esters, diisocyanate, glutaraldehyde, diazobenzenes, and hexamethylene diamines. Non-peptide linkers are well established in the art and may be suitable for the inducible IL-18 prodrugs described herein.

[0414] 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.

[0415] 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. B. Exemplary Inducible IL-18 Prodrugs

[0416] The inducible IL-18 prodrugs comprise an IL-18 polypeptide, a half-life extension element, an IL-18 blocking domain, and a protease cleavable linker. Preferred inducible IL-18 prodrugs comprise two polypeptide chains.

[0417] The inducible IL-18 prodrug can comprise: 1) a first polypeptide chain comprising: i) an IL-18 polypeptide chain that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 2- 170, 200-209, or 211-245, or an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 2-170, 200-209, or 211-245; ii) a half-life extension element that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 602-606, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-684; iii) a protease cleavable linker that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 354-379; and iv) an antigen binding portion of an antibody heavy chain that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 171, 173, 318, 320, 322, 331, 353, 354, 655-658, 734, 736, 738, 740, 742, 820, 821, 875, 920, 1005, 1006, 1010, 1011, or 1019 or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 171, 173, 318, 320, 322, 331, 353, 354, 655-658, 734, 736, 738, 740, 742, 820, 821, 875, 920, 1005, 1006, 1010, 1011, or 1019, in which the IL-18 polypeptide and the antigen binding portion of the antibody heavy chain and / or half-life extension element are operably linked by the protease cleavable linker; and 77 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 2) a second polypeptide comprising at least an antigen binding portion of an antibody light chain that is complementary to the heavy chain in the first polypeptide chain that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 172, 174, 187, 188, 312, 314, 319, 864, 865, 866, 867, 869, 870, 871, or 872, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 172, 174, 187, 188, 312, 314, 319, 864, 865, 866, 867, 869, 870, 871, or 872. The inducible IL-18 prodrug can comprise: 1) a first polypeptide chain comprising: i) an IL-18 polypeptide chain that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 2- 170, 200-209, or 211-245, or an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 2-170, 200-209, or 211-245; ii) a half-life extension element that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 602-606, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-684; iii) a protease cleavable linker that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 354-379; and iv) an antigen binding portion of an antibody light chain that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 172, 174, 187, 188, 312, 314, 319, 864, 865, 866, 867, 869, 870, 871, or 872 or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 172, 174, 187, 188, 312, 314, 319, 864, 865, 866, 867, 869, 870, 871, or 872, in which the IL-18 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 2) 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 chain that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 171, 173, 318, 320, 322, 331, 353, 354, 655-658, 734, 736, 738, 740, 742, 820, 821, 875, 920, 1005, 1006, 1010, 1011, or 1019, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 172, 171, 173, 318, 320, 322, 331, 353, 354, 655-658, 734, 736, 738, 740, 742, 820, 821, 875, 920, 1005, 1006, 1010, 1011, or 1019.

[0418] The inducible IL-18 prodrugs can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 175-186, 246-310, 332-345, 665-676, 678-696, 703- 732, 743-793, 798-811, 838-851, 878-880, 9191, 921-923, 925, 927-929, 940-949, 981-993, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NOs: 187, 188, 311-314, 315-317, 863-866, 869-872, or 1012-1016.

[0419] The inducible IL-18 prodrugs can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 324-327, 348, 677-683, 697-702, 794-797, 812-819, 824-837, 852-861, or 1021-1032, and 2) a second polypeptide that comprises or consists of the amino acid sequence of any one of SEQ IND NOs: 187, 311, 315, 316, 317, 868, 1013, 1015, 1017, or 1019. 78 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0420] The inducible IL-18 prodrugs can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 289, 1101, or 1103, and 2) a second polypeptide that comprises or consists of any one of SEQ ID NOs: 187, 1102, or 1104.

[0421] Compounds 1-77 are specific examples of inducible IL-18 prodrugs. Further details of exemplary inducible IL-18 prodrugs are described in Table 4. Compounds 78-276 are also specific examples of inducible IL-18 prodrugs. Further details of exemplary inducible IL-18 prodrugs are described in Table 4. Compounds 277-279 are further examples of inducible IL-18 prodrugs and details of these are described in Table 4. Table 4. Inducible IL-18 prodrugs Inducible IL-18 First Polypeptide Second Polypeptide Prodrug 79 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 21 SEQ ID NO: 254 SEQ ID NO: 312 Compound 22 SEQ ID NO: 255 SEQ ID NO: 311 80 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 54 SEQ ID NO: 287 SEQ ID NO: 313 Compound 55 SEQ ID NO: 288 SEQ ID NO: 313 81 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 89 SEQ ID NO: 345 SEQ ID NO: 863 Compound 90 SEQ ID NO: 665 SEQ ID NO: 864 82 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 122 SEQ ID NO: 711 SEQ ID NO: 870 Compound 123 SEQ ID NO: 712 SEQ ID NO: 870 83 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 155 SEQ ID NO: 754 SEQ ID NO: 870 Compound 156 SEQ ID NO: 755 SEQ ID NO: 188 84 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 188 SEQ ID NO: 786 SEQ ID NO: 187 Compound 189 SEQ ID NO: 787 SEQ ID NO: 187 85 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 221 SEQ ID NO: 849 SEQ ID NO: 314 Compound 222 SEQ ID NO: 850 SEQ ID NO: 314 86 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Compound 254 SEQ ID NO: 982 SEQ ID NO: 1015 Compound 255 SEQ ID NO: 983 SEQ ID NO: 1015

[0422] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 175, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187. comprises or consists of the amino acid sequence of SEQ ID NO: 176, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0424] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 177, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187. 87 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0425] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 178, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0426] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 179, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0427] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 180, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0428] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 181, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0429] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 182, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0430] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 183, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 188.

[0431] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 184, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 188.

[0432] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 185, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 188.

[0433] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 186, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 188.

[0434] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 250, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0435] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 257, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313. 88 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0436] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 259, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0437] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 260, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0438] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 261, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0439] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 262, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0440] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 287, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0441] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 288, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0442] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 289, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0443] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 290, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0444] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 297, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0445] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 298, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0446] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 246, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 311. 89 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0447] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 247, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 312.

[0448] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 248, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0449] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 249, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0450] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 250, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0451] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 250, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 312.

[0452] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 251, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 312.

[0453] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 252, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 312.

[0454] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 253, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 312.

[0455] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 254, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 312.

[0456] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 255, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 311.

[0457] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 256, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 311. 90 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0458] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 257, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0459] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 258, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0460] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 259, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0461] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 260, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0462] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 261, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0463] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 262, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0464] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 263, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0465] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 264, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0466] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 265, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0467] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 266, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0468] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 267, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315. 91 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0469] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 268, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0470] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 269, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0471] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 270, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0472] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 271, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0473] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 272, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0474] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 273, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0475] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 274, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0476] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 275, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0477] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 276, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0478] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 277, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0479] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 277, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315. 92 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0480] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 278, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0481] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 279, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0482] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 280, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0483] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 281, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 317.

[0484] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 282, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 317.

[0485] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 283, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 317.

[0486] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 284, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 317.

[0487] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 285, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 317.

[0488] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 286, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 317.

[0489] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 287, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0490] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 288, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313. 93 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0491] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 289, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0492] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 290, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0493] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 291, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0494] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 292, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0495] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 293, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0496] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 294, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313.

[0497] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 295, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0498] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 296, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0499] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 297, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0500] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 298, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0501] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 299, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314. 94 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0502] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 300, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0503] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 301, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0504] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 302, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0505] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 303, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0506] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 304, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0507] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 305, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 311.

[0508] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 306, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0509] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 307, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 315.

[0510] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 308, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0511] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 309, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 316.

[0512] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 310, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 313. 95 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0513] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1021, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0514] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1022, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0515] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1023, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0516] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1024, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0517] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1025, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0518] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1026, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0519] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1027, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0520] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1028, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0521] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1029, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0522] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1030, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0523] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1031, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314. 96 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320

[0524] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1032, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 314.

[0525] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1101, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1102.

[0526] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1103, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 187.

[0527] Inducible IL-18 prodrug can comprise 1) a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 289, and 2) a second polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 1104.

[0528] 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, or 78-276 that retain attenuated IL-18 activity in the periphery and that release active IL-18 upon protease cleavage in the tumor microenvironment can also be used in accordance with this disclosure. For example, the inducible IL-18 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: 175-186 and 246-310 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 SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 312, SEQ ID NO: 314, SEQ ID NO: 863, SEQ ID NO: 864, SEQ ID NO: 865, SEQ ID NO: 866, SEQ ID NO: 869, SEQ ID NO: 870, SEQ ID NO: 871, SEQ ID NO: 872, SEQ ID NO: 1012, SEQ ID NO: 1013, SEQ ID NO: 1014, SEQ ID NO: 1015 or SEQ ID NO: 1016.For example, the inducible IL-18 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: 332-345, 665-676, 678-696, 703-732, 743-793, 798-811, 838-851, 878-880, 9191, 921-923, 925, 927-929, 940-949, 981-993 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 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 97%, at least about 98%, or at least about 99% identical, to any one of SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 312, SEQ ID NO: 314, SEQ ID NO: 863, SEQ ID NO: 864, SEQ ID NO: 865, SEQ ID NO: 866, SEQ ID NO: 869, SEQ ID NO: 870, SEQ ID NO: 871, SEQ ID NO: 872, SEQ ID NO: 1012, SEQ ID NO: 1013, SEQ ID NO: 1014, SEQ ID NO: 1015 or SEQ ID NO: 1016.

[0529] Amino acid sequence variants of Compounds 277-279 that retain attenuated IL-18 activity in the periphery and that release active IL-18 upon protease cleavage in the tumor microenvironment can also be used in accordance with this disclosure. For example, the inducible IL-18 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 NO: 289, SEQ ID NO: 1101, or SEQ ID NO: 1103, 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 ID NO: 187, SEQ ID NO: 1102, or SEQ ID NO: 1104. E. Pharmaceutical Compositions

[0530] The disclosure further relates to pharmaceutical compositions comprising inducible IL-18 prodrugs as disclosed herein. Also provided herein are pharmaceutical formulations or compositions comprising the inducible IL-18 prodrugs described herein as described herein and typically a pharmaceutically acceptable carrier.

[0531] Compositions comprising the inducible IL-18 prodrugs 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.

[0532] 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 98 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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, 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-18 prodrugs described herein or nucleic acid sequences encoding the inducible IL-18 prodrugs described herein to humans or other subjects.

[0533] In some embodiments of the pharmaceutical compositions, inducible IL-18 prodrugs described herein is encapsulated in nanoparticles. In some embodiments, the nanoparticles are fullerenes, liquid crystals, liposome, quantum dots, superparamagnetic nanoparticles, dendrimers, or nanorods. In other embodiments of the pharmaceutical compositions, the inducible IL-18 prodrugs as described herein. In some instances, inducible IL-18 prodrugs described herein are conjugated to the surface of liposomes. In some instances, inducible IL-18 prodrugs described herein are encapsulated within the shell of a liposome. In some instances, the liposome is a cationic liposome.

[0534] The inducible IL-18 prodrugs described herein are suitable for use as a medicament and can be administered by an appropriate route of administration e.g. by intravenous, intraperitoneal, subcutaneous, intramuscular, topical or intradermal 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.

[0535] While parenteral administration is generally preferred, optionally, the inducible IL-18 prodrugs described herein, or nucleic acid sequences encoding the inducible IL-18 prodrugs 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 99 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 express and preferably secrete the encoded chimeric polypeptide or to therapeutically deliver nucleic acids to a subject.

[0536] 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 these 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 parenchyma, and a number of other tissue sites. Other useful systems include, for example, replicating and host-restricted non-replicating vaccinia virus vectors.

[0537] The inducible IL-18 prodrugs and / or nucleic acid molecules 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).

[0538] The inducible IL-18 prodrugs described 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.

[0539] 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 100 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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-18 prodrugs described herein by expression in a suitable host cell, such as CHO cells.

[0540] 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.

[0541] 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 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.

[0542] 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).

[0543] 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 101 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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.

[0544] If desired, an immune cell can be engineered to express a desired IL-18 polypeptide, or preferably a desired inducible IL-18 prodrug. For example, a T cell (e.g., TIL, CAR-T, TCR-T) can be engineered to express an inducible IL-18 prodrug as described herein. Similarly, other immune cells, such as NK cell can be engineered to express an inducible IL-18, and if desired, an antigen binding protein that directs NK cell activity to cells that express a selected antigen. Preferably, an inducible IL-18 prodrug is chosen such that its protease cleavable linker is cleaved by a protease with higher activity in the microenvironment of a tumor in comparison to other locations, and an immune cell capable of killing cells in the same tumor is chosen to express the inducible IL-18 prodrug. An immune cell may be engineered to express an inducible IL-18 prodrug using any suitable method, such as via one or more viral vectors. An immune cell engineered to express an inducible IL-18 prodrug may be autologous or allogeneic with respect to a subject receiving it as a therapy. In the case of an allogeneic cell, it may be advantageous to further engineer the cell such that it lacks functional endogenous TCRs, MHCs, or both. An immune cell may express inducible IL-18 prodrugs temporarily (such as if the immune cell is transduced with mRNA encoding the inducible IL-18 prodrug) or over the course of its lifespan (such as if the immune cell is transduced with DNA encoding the inducible IL-18 prodrug). An immune cell may be engineered to express an inducible IL-18 prodrug and an antigen binding protein (such as a CAR) by transducing it with a single polynucleotide encoding both the inducible IL-18 prodrug and the antigen binding protein. Alternatively, an immune cell may be engineered to express an inducible IL-18 prodrug and an antigen binding protein by transducing it with two or more polynucleotides, for example, one or more polynucleotide encoding the inducible IL-18 prodrug and one or more polynucleotide encoding the antigen binding protein.

[0545] In some embodiments, an immune cell expresses an inducible IL-18 prodrug as described herein. Generally, the inducible IL-18 prodrug comprises a IL-18 polypeptide, a protease cleavable linker, and a blocking element, as described herein. In some embodiments, a cell contains a single polynucleotide 102 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 encoding both an antigen binding protein and an inducible IL-18 prodrug as described herein. In some embodiments, a cell contains a first polynucleotide encoding an antigen binding protein and a second polynucleotide encoding an inducible IL-18 prodrug as described herein. In some embodiments, a cell contains a first polynucleotide encoding an antigen binding protein and a second polynucleotide encoding a inducible IL-18 prodrug comprising a IL-18 polypeptide, a protease cleavable linker, and a blocking element as described herein. Preferably, an antigen binding protein expressed by an immune cell directs the immune cell and its immune effector functions to cells that express a desired antigen. In some embodiments, a cell contains one or more polynucleotides encoding the inducible IL-18 prodrug and one or more polynucleotides encoding the antigen binding protein. The polynucleotides encoding the antigen binding protein and / or the inducible IL-18 prodrug are typically engineered for expression in a desired cell and are introduced into the cell using suitable methods, e.g., transduction, transfection. Accordingly, such polynucleotides are not found in the naturally occurring cell type or species, e.g., human. For example, an immune cell can contain at least three or more polynucleotides that collectively encode the antigen binding protein and the inducible IL-18 prodrug. The immune cell may contain two or more polynucleotides encoding an inducible IL-18 prodrug and at least one polypeptide encoding an antigen binding protein.

[0546] The inducible IL-18 prodrug can be expressed by the cell as a soluble protein, which is secreted by the cell into the extracellular space. The inducible IL-18 prodrug can also be expressed by the cell as a membrane-associated protein, which can optionally be removed from the membrane by proteolytic cleavage. Methods for designing membrane-associated proteins are well-known in the art and include inducible IL-18 prodrugs that include a suitable transmembrane or membrane anchoring sequence, and optionally an intracellular domain that is fused to the inducible IL-18 prodrug, optionally through a suitable spacer sequence. The spacer sequence can include a cleavage site for a protease that has greater activity in the tumor microenvironment than in other locations. Upon cleavage of such a spacer sequence, the IL-18 or inducible IL-18 prodrug can be released from the cell membrane. Suitable spacer sequences include the linkers disclosed herein, and amino acid sequences that include cleavage sites disclosed herein. Suitable transmembrane or membrane anchoring sequences are well-known in the art and include the transmembrane segments of a platelet derived growth factor receptor (PDGFR,e.g., PDGFRbeta, PDGFRalpha), CD4, CD8, IL-18R and the like. Preferably, the membrane associate inducible IL-18 prodrug lacks a cytoplasmic portion or has a cytoplasmic portion that does not transduce activation signals to the engineered cell. In one example, a CAR-T cell also expresses an inducible IL-18 prodrug that can be a secreted molecule or membrane-associated. The engineered cell can be a T cell that expresses an inducible IL-18 prodrug. Preferably the engineered T cell is a TIL or a CAR-T cell. The inducible IL-18 prodrug can be soluble or membrane tethered. An inducible IL-18 prodrug that is 103 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 membrane tethered can comprise a IL-18 polypeptide, a first protease cleavable linker, and a blocking element as described herein. The inducible IL-18 prodrug can contain a suitable transmembrane domain or membrane anchoring sequence. Typically, the suitable transmembrane domain or membrane anchoring sequence is linked to the IL-18 peptide, directly or indirectly, through a protease cleavable linker and can be released from the membrane upon cleavage of such protease cleavable linker. For example, a membrane tethered inducible IL-18 prodrug can comprise from amino to carboxy terminus: the IL-18 polypeptide – a protease cleavable linker – a blocking element (e.g., a scFv, a VH and CH1, or VL and CL) – a cleavable linker that is preferably protease cleavable – a suitable transmembrane or membrane anchoring sequence. As another example, a membrane tethered inducible IL-18 prodrug can comprise from amino to carboxy terminus: a blocking element (e.g., e.g., a scFv, a VH and CH1, or VL and CL) – a protease cleavable linker – the IL-18 polypeptide – a cleavable linker that is preferably protease cleavable – a suitable transmembrane or membrane anchoring sequence. In some preferred embodiments, the inducible IL-18 prodrug is membrane tethered and comprises the transmembrane domain of PDGFR, CD4, CD8 or IL-18R.

[0547] The compositions disclosed herein can comprise nucleic acids encoding the inducible IL-18 prodrugs described herein. Nucleic acids include DNA and RNA molecules. Exemplary RNA molecules include, but are not limited to mRNA, siRNA, circular RNA, self-replication RNA, transfer RNA, small RNA, miRNA, short hairpin RNAs, double-stranded RNAs, long noncoding RNAs, small cell nuclear RNAs (Y NRA), short interfering RNAs (siRNA), antisense oligonucleotide (ASO), messenger RNA (mRNA), or guide RNA on a ribonucleoprotein (RNP).

[0548] The composition can comprise DNA encoding the inducible IL-18 prodrugs described herein, or the fusion polypeptides or conjugates described herein. The composition can comprise RNA encoding the inducible IL-18 prodrugs described herein, or the fusion polypeptides or conjugates described herein. In particular, the composition can comprise mRNA encoding the inducible IL-18 prodrugs described herein, or the fusion polypeptides or conjugates described herein. The composition can comprise an RNA sequence (e.g., mRNA) transcribed from a DNA sequence encoding the inducible IL-18 prodrugs described herein, or the fusion polypeptides or conjugates described herein.

[0549] The RNA can comprise a 5’ untranslated region (UTR). The 5’ UTR can be upstream of an initiation codon. Without wishing to be bound by theory or mechanism, the 5’UTR can regulate translation of RNA, stabilizes the RNA, and / or increases the half-life of the RNA. Suitable 5’ UTRs are well-known in the art. The RNA can comprise a 3’ untranslated region (UTR). The 3’ UTR can follow the translation termination codon. Without wishing to be bound by theory or mechanism, the 3’ UTR can regulate polyadenylation, translation efficiency, localization, or stability of the RNA. Suitable 3’ UTRs 104 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 are well-known in the art. The composition generally comprises both a 5’ UTR and a 3’ UTR. However, if desired the composition can comprise only a 5’ UTR or only a 3’ UTR.

[0550] The RNA can comprise a poly-A tail. The poly-A tail can be at least about 25 nucleotides, at least about 30 nucleotides, at least about 40 nucleotides, at least about 50 nucleotides, at least about 70 nucleotides, at least about 80 nucleotides, at least about 90 nucleotides, at least about 100 nucleotides, or more. Suitable poly-A tails are well known in the art.

[0551] In certain embodiments, the RNA can comprise a 5’ cap, a 5’UTR, a nucleic acid encoding the inducible IL-18 prodrugs disclosed herein, or the fusion polypeptides or conjugates described herein, a 3’ UTR, and a poly-A tail.

[0552] The nucleic acids and compositions disclosed herein can comprise any desired modification. For example, one or more nucleosides can be modified or one or more ribose can be modified. The modifications can improve stability of the nucleic acids and compositions disclosed herein. Suitable modifications are well-known in the art.

[0553] For instance, one or more uridine in the RNA can be replaced by a modified nucleoside. The modified nucleoside can be a modified uridine. The modified uridine replacing uridine is pseudouridine (ψ), N1-methyl-pseudouridine (m1ψ), or 5-methyl-uridine (m5U).

[0554] One or more cytosine, adenine or guanine in the RNA can be replaced by modified nucleobase(s). The modified nucleobase replacing cytosine can be 5-methylcytosine (m5C). The modified nucleobase replacing adenine can be N6-methyladenine (m6A). Any other modified nucleobase known in the art for reducing the immunogenicity of the molecule can be used.

[0555] The modified nucleoside replacing one or more uridine in the RNA can include, but is not limited to, one or more of 3-methyl-uridine (m3U), 5-methoxy-uridine (mo5U), 5-aza-uridine, 6-aza- uridine, 2- thio-5-aza-uridine, 2-thio-uridine (s2U), 4-thio-uridine (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho5U), 5-aminoallyl-uridine, 5-halo-uridine (e.g., 5- iodo-uridineor 5-bromo-uridine), uridine 5-oxyacetic acid (cmo5U), uridine 5-oxyacetic acid methyl ester (mcmo5U), 5-carboxymethyl- uridine (cm5U), 1-carboxymethyl-pseudouridine, 5-carboxyhydroxymethyl-uridine (chm5U), 5- carboxyhydroxymethyl-uridine methyl ester (mchm5U), 5-methoxycarbonylmethyl-uridine (mcm5U), 5- methoxycarbonylmethyl-2-thio- uridine (mcm5s2U), 5-aminomethyl-2-thio-uridine (nm5s2U), 5- methylaminomethyl-uridine (mnm5U), 1-ethyl-pseudouridine, 5-methylaminomethyl-2-thio-uridine (mnm5s2U), 5- methylaminomethyl-2-seleno-uridine (mnm5se2U), 5-carbamoylmethyl-uridine (ncm5U), 5- carboxymethylaminomethyl-uridine (cmnm5U), 5-carboxymethylaminomethyl-2-thio-uridine (cmnm5s2U), 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyl-uridine (τm5U), 1- taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine(τm5s2U), 1-taurinomethyl-4- thio- pseudouridine), 5-methyl-2-thio-uridine (m5s2U), 1-methyl-4-thio-pseudouridine (m1s4ψ), 4-thio-1- 105 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 methyl-pseudouridine, 3-methyl-pseudouridine (m3ψ), 2-thio-1-methyl- pseudouridine, 1-methyl-1-deaza- pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine (D), dihydropseudouridine, 5,6- dihydrouridine, 5-methyl-dihydrouridine (m5D), 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2- methoxy-uridine, 2-methoxy- 4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl- pseudouridine, 3-(3-amino-3-carboxypropyl)uridine (acp3U), 1-methyl-3-(3-amino-3- carboxypropyl)pseudouridine (acp3ψ), 5-(isopentenylaminomethyl)uridine (inm5U), 5- (isopentenylaminomethyl)-2-thio-uridine (inm5s2U), α-thio-uridine, 2′-O-methyl-uridine (Um), 5,2′-O- dimethyl-uridine (m5Um), 2′-O-methyl-pseudouridine (ψm), 2-thio-2′-O- methyl-uridine (s2Um), 5- methoxycarbonylmethyl-2′-O-methyl-uridine (mcm5Um), 5- carbamoylmethyl-2′-O-methyl-uridine (ncm5Um), 5-carboxymethylaminomethyl-2′-O- methyl-uridine (cmnm5Um), 3,2′-O-dimethyl-uridine (m3Um), 5-(isopentenylaminomethyl)- 2′-O-methyl-uridine (inm5Um), 1-thio-uridine, deoxythymidine, 2′-F-ara-uridine, 2′-F- uridine, 2′-OH-ara-uridine, 5-(2-carbomethoxyvinyl) uridine, 5-[3-(1-E- propenylamino)uridine, or any other modified uridine known in the art.

[0556] The compositions disclosed herein can comprising one or more nucleic acids (e.g. DNA or RNA) as described herein. For example, the composition can comprise one RNA, two RNAs, three RNAs, four RNAs, five RNAs, or more. F. Therapeutic Uses

[0557] 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-18 prodrug as described herein. Diseases, disorders, or conditions include, but are not limited to, cancer, inflammatory disease, an immunological disorder, autoimmune disease, infectious disease (i.e., bacterial, viral, or parasitic disease). Preferably, the disease, disorder, or condition is cancer.

[0558] Any suitable cancer may be treated with the inducible IL-18 prodrugs described herein. Illustrative suitable cancers, in particular solid tumors, such as sarcomas and carcinomas. For examples, 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, 106 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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.

[0559] In certain embodiments, the methods and compositions disclosed herein can be used to treat 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, 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, 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, mycosis fungoides, 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 107 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 carcinoma, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, non-Hodgkin lymphoma, squamous carcinoma of the head and neck, malignant pleural mesothelioma, and Wilms tumor.

[0560] In certain preferred 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.

[0561] 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).

[0562] Preferably, the methods and compositions disclosed herein are used to treat colon cancer, lung cancer, melanoma, renal cell carcinoma, or breast cancer.

[0563] 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.

[0564] 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-18 prodrug described herein to the subject. The enhanced immune response may prevent, delay, or treat the onset of cancer, a tumor, or a viral disease. In some embodiments, the methods described herein increase the activity of Natural Killer Cells and T lymphocytes.

[0565] The method can further involve the administration of one or more additional agents to treat cancer, such as chemotherapeutic agents (e.g., Adriamycin, Cerubidine, Bleomycin, Alkeran, Velban, Oncovin, Fluorouracil, Thiotepa, Methotrexate, Bisantrene, Noantrone, Thiguanine, Cytaribine, 108 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 Procarabizine), immuno-oncology agents (e.g., anti-PD-L1, anti-CTLA4, anti-PD-1, anti-LAG3, anti- CD47, anti-GD2), cellular therapies (e.g., CAR-T, T-cell therapy), oncolytic viruses and the like. Non- limiting examples of anti-cancer agents that can be used include acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantrone acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; ilmofosine; interleukin II (including recombinant interleukin II, or rIL2), interferon alpha-2a; interferon alpha-2b; interferon alpha-nl interferon alpha-n3; interferon beta-I; interferon gamma-I b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestolone acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; 109 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinzolidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicin hydrochloride.

[0566] In some embodiments of the methods described herein, the s inducible IL-18 prodrugs described herein is administered in combination with an agent for the treatment of the particular disease, disorder, or condition. Agents include, but are not limited to, therapies involving antibodies, small molecules (e.g., chemotherapeutics), hormones (steroidal, peptide, and the like), radiotherapies (γ-rays, C-rays, and / or the directed delivery of radioisotopes, microwaves, UV radiation and the like), gene therapies (e.g., antisense, retroviral therapy and the like) and other immunotherapies. In some embodiments, the inducible IL-18 prodrugs described herein or is administered in combination with anti-diarrheal agents, anti-emetic agents, analgesics and / or non-steroidal anti-inflammatory agents.

[0567] This disclosure relates to a therapeutic combination of any of the sdAbs as described herein, or a inducible IL-18 prodrug described herein in combination with one or more additional agents to treat cancer (such as lymphoma), such as chemotherapeutic agents (e.g., cyclophosphamide, mechlorethamine, melphalan, chlorambucil, ifosfamide, busulfan, N-Nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, streptozotocin, dacarbazine, mitozolomide, temozolomide, thiotepa, mitomycin, diaziquone (AZQ), cisplatin, carboplatin, oxaliplatin, procarbazine, hexamethylmelamine, methotrexate, pemetrexed, fluorouracil (e.g.5-fluorouracil), capecitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, pentostatin, thioguanine, mercaptopurine, vincristine, vinblastine, vinorelbine, vindesine, vinflunine, paclitaxel, docetaxel, etoposide, teniposide, doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, mitoxantrone, actinomycin, bleomycin, bisantrene, gemcitabine, cytarabine, and the like), immuno-oncology agents and immune checkpoint inhibitors (e.g., anti-PD-L1, anti-CTLA4, anti-PD-1, anti-LAG3, anti-CD47, anti-GD2), oncolytic viruses and the like.

[0568] The inducible IL-18 prodrugs described herein can be combined with any desired additional anti- cancer agent. The s inducible IL-18 prodrugs described herein can be combined with any desired anti-PD- 1 antibody or any desired anti-PD-L1 antibody.

[0569] Exemplary anti-PD-1 antibodies that can be combined with the sdAbs as described herein, or a inducible IL-18 prodrug described herein, 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), 110 \\4127-7386-4278 v1 Attorney Docket No.: 761146.362320 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 Biologi...

Claims

Attorney Docket No.: 761146.362320 CLAIMS 1. An inducible IL-18 prodrug comprising at least one of each of: a) an IL-18 polypeptide [A], wherein the IL-18 polypeptide comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245; b) a half-life extension element [H]; c) an IL-18 blocking element [D]; and d) a protease cleavable linker [L]; wherein the IL-18 polypeptide and the IL-18 blocking element or the half-life extension element are operably linked by the protease-cleavable polypeptide linker and the inducible IL-18 prodrug has attenuated IL-18 receptor activating activity, wherein the IL-18 receptor activating activity of the inducible IL-18 prodrug is at least about 10X less than the IL-18 receptor activating activity of the polypeptide that contains the IL-18 polypeptide that is produced by cleavage of the protease cleavable linker.

2. The inducible IL-18 prodrug of claim 1, wherein the inducible IL-18 prodrug has the formula: [A]-[L1]-[H]-[L2]-[D] [D]-[L2]-[H]-[L1]-[A] [A]-[L1]-[D]-[L2]-[H] [H]-[L2]-[D]-[L1]-[A] [H]-[L1]-[A]-[L2’]-[D] [D]-[L1]-[A]-[L2’]-[H] wherein [A] is an IL-18 polypeptide, [D] is an IL-18 blocking element, [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 [L2’] is a protease-cleavable polypeptide linker.

3. An inducible IL-18 prodrug comprising an IL-18 polypeptide, a half-life extension element, an IL-18 blocking element and a protease cleavable linker, wherein the IL-18 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-18 prodrug comprises: i. a first polypeptide comprising the IL-18 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-18 polypeptide and the antigen binding portion of the antibody 269 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 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; wherein the IL-18 polypeptide comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245; 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-18 binding site.

4. The inducible IL-18 prodrug of any one of the preceding claims, wherein the half-life extension element comprises a serum albumin binding domain, a serum albumin, transferrin, or immunoglobulin Fc, or fragment thereof.

5. The inducible IL-18 prodrug of claim 4, wherein the half-life extension element is a serum albumin binding domain.

6. The inducible IL-18 prodrug of claim 5, wherein the serum albumin binding domain is a single domain antibody fragment (sdAb).

7. The inducible IL-18 prodrug of claim 6, wherein the sdAb comprises the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-654, an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-654.

8. The inducible IL-18 prodrug of any one of claims 1 or 2, wherein the half-life extension element comprises the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-654, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-684.

9. The inducible IL-18 prodrug of any one of the preceding claims, wherein the IL-18 blocking domain comprises a ligand-binding domain or fragment of a cognate receptor for the cytokine polypeptide, or an antibody or antigen-binding fragment of an antibody that binds to the cytokine polypeptide. 270 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 10. The inducible IL-18 prodrug of claim 9, wherein the antibody or antigen-binding fragment is a single domain antibody, a Fab, or a scFv that binds the cytokine polypeptide.

11. The inducible IL-18 prodrug of claim 10, wherein the antibody or antigen-binding fragment is an scFv.

12. The inducible IL-18 prodrug of claim 10, wherein the antibody or antigen-binding fragment is a Fab.

13. The inducible IL-18 prodrug of any one of the preceding claims, wherein the blocking element comprises Blocking Element 1 (SEQ ID NO: 1033 / SEQ ID NO: 1058), Blocking Element 2 (SEQ ID NO: 1034 / SEQ ID NO: 1059), Blocking Element 3 (SEQ ID NO: 1035 / SEQ ID NO: 1060), Blocking Element 4 (SEQ ID NO: 1036 / SEQ ID NO: 1061), Blocking Element 5 (SEQ ID NO: 1037 / SEQ ID NO: 1062), Blocking Element 6 (SEQ ID NO: 1038 / SEQ ID NO: 1063), Blocking Element 7 (SEQ ID NO: 1039 / SEQ ID NO: 1064), Blocking Element 8 (SEQ ID NO: 1040 / SEQ ID NO: 1065), Blocking Element 9 (SEQ ID NO: 1041 / SEQ ID NO: 1066), Blocking Element 10 (SEQ ID NO: 1042 / SEQ ID NO: 1067), Blocking Element 11 (SEQ ID NO: 1043 / SEQ ID NO: 1068), Blocking Element 12 (SEQ ID NO: 1044 / SEQ ID NO: 1069), Blocking Element 13 (SEQ ID NO: 1045 / SEQ ID NO: 1070), Blocking Element 14 (SEQ ID NO: 1046 / SEQ ID NO: 1071), Blocking Element 15 (SEQ ID NO: 1047 / SEQ ID NO: 1072), Blocking Element 16 (SEQ ID NO: 1048 / SEQ ID NO: 1073), Blocking Element 17 (SEQ ID NO: 1049 / SEQ ID NO: 1074), Blocking Element 18 (SEQ ID NO: 1050 / SEQ ID NO: 1075), Blocking Element 19 (SEQ ID NO: 1051 / SEQ ID NO: 1076), Blocking Element 20 (SEQ ID NO: 1052 / SEQ ID NO: 1077), Blocking Element 21 (SEQ ID NO: 1053 / SEQ ID NO: 1078), Blocking Element 22 (SEQ ID NO: 1054 / SEQ ID NO: 1079), Blocking Element 23 (SEQ ID NO: 1055 / SEQ ID NO: 1080), Blocking Element 24 (SEQ ID NO: 1056 / SEQ ID NO: 1081), Blocking Element 25 (SEQ ID NO: 1057 / SEQ ID NO: 1082).

14. The inducible IL-18 prodrug of claim 13, wherein the blocking element further comprises a heavy chain constant region that comprises or consists of SEQ ID NO: 1083 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 1083, and a light chain constant region that comprises or consists of SEQ ID NO: 1084 or an amino acid sequence that has at least 90% identity to SEQ ID NO: 1084.

15. The inducible IL-18 prodrug of any one of claims 1, 2, 10, 13, or 14, wherein the antibody or antigen-binding fragment comprises an antibody light chain (VL + CL) comprising the amino acid 271 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 sequence of SEQ ID NO: 172, SEQ ID NO: 174, or SEQ ID NO: 322 and an antibody heavy chain Fab fragment (VH + CH1) comprising the amino acid sequence of SEQ ID NO: 171, SEQ ID NO: 173, or SEQ ID NO:

323.

16. The inducible IL-18 prodrug of any one of claims 1, 2, 10, 13, or 14, wherein the blocking element comprises an antibody light chain (VL + CL) comprising the amino acid sequence of SEQ ID NO: 172, SEQ ID NO: 174, or SEQ ID NO: 322, and antibody heavy chain Fab fragment (VH + CH1) comprising the amino acid sequence of SEQ ID NO: 171, SEQ ID NO: 173, or SEQ ID NO:

322.

17. The inducible IL-18 prodrug of any one of claims 1, 2, 10, 13, or 14, wherein the blocking element comprises 1) a first polypeptide chain that comprises a first polypeptide 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 and 2) a second polypeptide that comprises or consists of SEQ ID NO: 172, or an amino acid sequence that has at least 95% identity to SEQ ID NO:

172.

18. The blocking element can of any one of claims 1, 2, 10, 13, or 14, wherein the blocking element comprises 1) a first polypeptide chain that comprises a first polypeptide 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 and 2) a second polypeptide that comprises or consists of SEQ ID NO: 174, or an amino acid sequence that has at least 95% identity to SEQ ID NO:

174.

19. The blocking element can of any one of claims 1, 2, 10, 13, or 14, wherein the blocking element comprises 1) a first polypeptide chain that comprises a first polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 322, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 322 and 2) a second polypeptide that comprises or consists of SEQ ID NO: 323, or an amino acid sequence that has at least 95% identity to SEQ ID NO:

323.

20. The inducible IL-18 prodrug of any one of the preceding claims, wherein the cytokine blocking domain inhibits activation of the cytokine polypeptide receptor by the inducible cytokine prodrug.

21. The inducible IL-18 prodrug of any one of the preceding claims, wherein the protease cleavable linker comprises a sequence that is capable of being cleaved by a protease selected from kallikrein, thrombin, chymase, carboxypeptidase A, cathepsin, elastase, PR-3, granzyme M, a calpain, a matrix 272 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 metalloproteinase (MMP), an ADAM, a FAP, a plasminogen activator, a caspase, a tryptase, or a tumor protease.

22. The inducible IL-18 prodrug of claim 20, wherein the protease is selected from cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, cathepsin G, MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, or MMP14.

23. The inducible IL-18 prodrug of any one of the preceding claims, wherein the protease cleavable linker comprises at least two sequences that are independently capable of being cleaved by a protease.

24. The inducible IL-18 prodrug of any one of the preceding claims, wherein the protease cleavable linker comprises a synthetic sequence.

25. The inducible IL-18 prodrug of any one of the preceding claims, wherein the protease cleavable linker comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 355-379 or 426, or an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of 355-379 or 426.

26. The inducible IL-18 prodrug of claim 25, wherein the protease cleavable linker comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 355-379 or 426.

27. The inducible IL-18 prodrug of claim 26, wherein the protease cleavable linker comprises an amino acid sequence of SEQ ID NO:

426.

28. The inducible IL-18 prodrug of claim 26, wherein the protease cleavable linker comprises an amino acid sequence of SEQ ID NO:

357.

29. The inducible IL-18 prodrug of any one of the preceding claims, wherein the inducible IL-18 prodrug comprises any one of Compounds 1-264 as shown in Table 4.

30. The inducible IL-18 prodrug of any one of the preceding claims, wherein the inducible IL-18 prodrug comprises a 1) a first polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 175-186, 246-310, 332-345, 665-676, 678-696, 703-732, 743-793, 798-811, 838- 851, 878-880, 9191, 921-923, 925, 927-929, 940-949, 981-993, and 2) a second polypeptide that 273 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 comprises or consists of the amino acid sequence of any one of SEQ IND NOs: 187, 188, 311-314, 315- 317, 863-866, 869-872, or 1012-1016.

31. The inducible IL-18 prodrug of any one of the preceding claims, wherein the inducible IL-18 prodrug comprises Compound 1 (SEQ ID NO: 175 / SEQ ID NO: 187), Compound 2 (SEQ ID NO: 176 / SEQ ID NO: 187), Compound 3 (SEQ ID NO: 177 / SEQ ID NO: 187), Compound 4 (SEQ ID NO: 178 / SEQ ID NO: 187), Compound 5 (SEQ ID NO: 179 / SEQ ID NO: 187), Compound 6 (SEQ ID NO: 180 / SEQ ID NO: 187), Compound 7 (SEQ ID NO: 181 / SEQ ID NO: 187), Compound 8 (SEQ ID NO: 182 / SEQ ID NO: 187), Compound 9 (SEQ ID NO: 183 / SEQ ID NO: 188), Compound 10 (SEQ ID NO: 184 / SEQ ID NO: 188), Compound 11 (SEQ ID NO: 185 / SEQ ID NO: 188), or Compound 12 (SEQ ID NO: 186 / SEQ ID NO: 188), Compound 13 (SEQ ID NO: 246 / SEQ ID NO: 311), Compound 14 (SEQ ID NO: 247 / SEQ ID NO: 312), Compound 15 (SEQ ID NO: 248 / SEQ ID NO: 312), Compound 16 (SEQ ID NO: 249 / SEQ ID NO: 314), Compound 17 (SEQ ID NO: 250 / SEQ ID NO: 313), Compound 18 (SEQ ID NO: 251 / SEQ ID NO: 313), Compound 19 (SEQ ID NO: 252 / SEQ ID NO: 313), Compound 20 (SEQ ID NO: 253 / SEQ ID NO: 313), Compound 21 (SEQ ID NO: 254 / SEQ ID NO: 313), Compound 22 (SEQ ID NO: 255 / SEQ ID NO: 311), Compound 23 (SEQ ID NO: 256 / SEQ ID NO: 311), Compound 24 (SEQ ID NO: 257 / SEQ ID NO: 313), Compound 25 (SEQ ID NO: 258 / SEQ ID NO: 313), Compound 26 (SEQ ID NO: 259 / SEQ ID NO: 314), Compound 27 (SEQ ID NO: 260 / SEQ ID NO: 314), Compound 28 (SEQ ID NO: 261 / SEQ ID NO: 314), Compound 29 (SEQ ID NO: 262 / SEQ ID NO: 314), Compound 30 (SEQ ID NO: 263 / SEQ ID NO: 313), Compound 31 (SEQ ID NO: 264 / SEQ ID NO: 313), Compound 32 (SEQ ID NO: 265 / SEQ ID NO: 315), Compound 33 (SEQ ID NO: 266 / SEQ ID NO: 315), Compound 34 (SEQ ID NO: 267 / SEQ ID NO: 315), Compound 35 (SEQ ID NO: 268 / SEQ ID NO: 315), Compound 36 (SEQ ID NO: 269 / SEQ ID NO: 316), Compound 37 (SEQ ID NO: 270 / SEQ ID NO: 316), Compound 38 (SEQ ID NO: 271 / SEQ ID NO: 316), Compound 39 (SEQ ID NO: 272 / SEQ ID NO: 316), Compound 40 (SEQ ID NO: 273 / SEQ ID NO: 315), Compound 41 (SEQ ID NO: 274 / SEQ ID NO: 315), Compound 42 (SEQ ID NO: 275 / SEQ ID NO: 315), Compound 43 (SEQ ID NO: 276 / SEQ ID NO: 315), Compound 44 (SEQ ID NO: 277 / SEQ ID NO: 315), Compound 45 (SEQ ID NO: 278 / SEQ ID NO: 315), Compound 46 (SEQ ID NO: 279 / SEQ ID NO: 315), Compound 47 (SEQ ID NO: 280 / SEQ ID NO: 315), Compound 48 (SEQ ID NO: 281 / SEQ ID NO: 317), Compound 49 (SEQ ID NO: 282 / SEQ ID NO: 317), Compound 50 (SEQ ID NO: 283 / SEQ ID NO: 317), Compound 51 (SEQ ID NO: 284 / SEQ ID NO: 317), Compound 52 (SEQ ID NO: 285 / SEQ ID NO: 317), Compound 53 (SEQ ID NO: 286 / SEQ ID NO: 317), Compound 54 (SEQ ID NO: 287 / SEQ ID NO: 313), Compound 55 (SEQ ID NO: 288 / SEQ ID NO: 313), Compound 56 (SEQ ID NO: 289 / SEQ ID NO: 313), Compound 57 (SEQ ID NO: 290 / SEQ ID NO: 313), Compound 58 (SEQ ID NO: 291 / SEQ ID NO: 313), Compound 59 (SEQ ID NO: 292 / SEQ ID NO: 313), Compound 60 (SEQ ID 274 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 NO: 293 / SEQ ID NO: 313), Compound 61 (SEQ ID NO: 294 / SEQ ID NO: 313), Compound 62 (SEQ ID NO: 295 / SEQ ID NO: 314), Compound 63 (SEQ ID NO: 296 / SEQ ID NO: 314), Compound 64 (SEQ ID NO: 297 / SEQ ID NO: 314), Compound 65 (SEQ ID NO: 298 / SEQ ID NO: 314), Compound 66 (SEQ ID NO: 299 / SEQ ID NO: 314), Compound 67 (SEQ ID NO: 300 / SEQ ID NO: 314), Compound 68 (SEQ ID NO: 301 / SEQ ID NO: 316), Compound 69 (SEQ ID NO: 302 / SEQ ID NO: 316), Compound 70 (SEQ ID NO: 303 / SEQ ID NO: 316), Compound 71 (SEQ ID NO: 304 / SEQ ID NO: 316), Compound 72 (SEQ ID NO: 305 / SEQ ID NO: 311), Compound 73 (SEQ ID NO: 306 / SEQ ID NO: 315), Compound 74 (SEQ ID NO: 307 / SEQ ID NO: 315), Compound 75 (SEQ ID NO: 308 / SEQ ID NO: 316), Compound 76 (SEQ ID NO: 309 / SEQ ID NO: 316), or Compound 77 (SEQ ID NO: 310 / SEQ ID NO: 313).

32. A nucleic acid encoding the inducible IL-18 prodrug of any one of claims 1-31.

33. The nucleic acid of claim 32, wherein the nucleic acid comprises a circular vector.

34. The nucleic acid of claim 32, wherein the nucleic acid comprises DNA.

35. The nucleic acid of claim 32, wherein the nucleic acid comprises RNA.

36. A vector comprising the nucleic acid of claim 32-35.

37. A host cell comprising the vector of claim 36.

38. A method of making a pharmaceutical composition, comprising culturing the host cell of claim 37 under suitable conditions for expression of the inducible IL-18 prodrug.

39. A pharmaceutical composition comprising the inducible IL-18 prodrug of any one of claims 1-31, or nucleic acid of any one of claims 32-36.

40. A method of using the inducible IL-18 prodrug of any one of claims 1-31 comprising administering an effective amount of the inducible cytokine prodrug to a subject in need thereof.

41. A method for treating cancer, comprising administering to a subject in need thereof an effective amount of the inducible IL-18 prodrug of any one of claims 1-31. 275 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 42. A inducible IL-18 prodrug of any one of claims 1-31, wherein the inducible IL-18 prodrug can be selected from the group consisting of Compounds 1-77.

43. A inducible IL-18 prodrug of any one of claims 1-31, wherein the inducible IL-18 prodrug can be selected from the group consisting of Compounds 78-264.

44. An inducible IL-18 prodrug comprising at least one of each of: a) an IL-18 polypeptide [A], wherein the IL-18 polypeptide comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245; b) a half-life extension element [H], wherein the half-life extension element comprises any one of SEQ ID NOs: 602-606 and 608-654, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-684; c) an IL-18 blocking element [D]; and d) a protease cleavable linker [L], wherein the protease cleavable linker comprises any one of SEQ ID NOs: 355-379 or 426 or an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of 355-379 or 426; wherein the IL-18 polypeptide and the IL-18 blocking element or the half-life extension element are operably linked by the protease-cleavable polypeptide linker and the inducible IL-18 prodrug has attenuated IL-18 receptor activating activity, wherein the IL-18 receptor activating activity of the inducible IL-18 prodrug is at least about 10X less than the IL-18 receptor activating activity of the polypeptide that contains the IL-18 polypeptide that is produced by cleavage of the protease cleavable linker.

45. The inducible IL-18 prodrug of claim 44, wherein the IL-18 blocking element is selected from the group consisting of: i) a first polypeptide chain that comprises a first polypeptide that comprises the amino acid sequence of SEQ ID NO: 171 and a second polypeptide that comprises of SEQ ID NO: 172, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 172; ii) a first polypeptide chain that comprises a first polypeptide that comprises the amino acid sequence of SEQ ID NO: 173, and a second polypeptide that comprises of SEQ ID NO: 174, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 174; iii) a first polypeptide chain that comprises a first polypeptide that comprises the amino acid sequence of SEQ ID NO: 318, and a second polypeptide that comprises of SEQ ID NO: 319, or an amino acid sequence that has at least 95% identity to SEQ ID NO: 319; or 276 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 iv) a first polypeptide chain that comprises a first polypeptide that comprises the amino acid sequence of SEQ ID NO: 320, and a second polypeptide that comprises SEQ ID NO: 321; and v) a first polypeptide chain that comprises the amino acid sequence of SEQ ID NO: 322, and a second polypeptide that comprises SEQ ID NO:

323.

46. The inducible IL-18 prodrug of claim 44, wherein the blocking element is selected from: Blocking Element 1 (SEQ ID NO: 1033 / SEQ ID NO: 1058), Blocking Element 2 (SEQ ID NO: 1034 / SEQ ID NO: 1059), Blocking Element 3 (SEQ ID NO: 1035 / SEQ ID NO: 1060), Blocking Element 4 (SEQ ID NO: 1036 / SEQ ID NO: 1061), Blocking Element 5 (SEQ ID NO: 1037 / SEQ ID NO: 1062), Blocking Element 6 (SEQ ID NO: 1038 / SEQ ID NO: 1063), Blocking Element 7 (SEQ ID NO: 1039 / SEQ ID NO: 1064), Blocking Element 8 (SEQ ID NO: 1040 / SEQ ID NO: 1065), Blocking Element 9 (SEQ ID NO: 1041 / SEQ ID NO: 1066), Blocking Element 10 (SEQ ID NO: 1042 / SEQ ID NO: 1067), Blocking Element 11 (SEQ ID NO: 1043 / SEQ ID NO: 1068), Blocking Element 12 (SEQ ID NO: 1044 / SEQ ID NO: 1069), Blocking Element 13 (SEQ ID NO: 1045 / SEQ ID NO: 1070), Blocking Element 14 (SEQ ID NO: 1046 / SEQ ID NO: 1071), Blocking Element 15 (SEQ ID NO: 1047 / SEQ ID NO: 1072), Blocking Element 16 (SEQ ID NO: 1048 / SEQ ID NO: 1073), Blocking Element 17 (SEQ ID NO: 1049 / SEQ ID NO: 1074), Blocking Element 18 (SEQ ID NO: 1050 / SEQ ID NO: 1075), Blocking Element 19 (SEQ ID NO: 1051 / SEQ ID NO: 1076), Blocking Element 20 (SEQ ID NO: 1052 / SEQ ID NO: 1077), Blocking Element 21 (SEQ ID NO: 1053 / SEQ ID NO: 1078), Blocking Element 22 (SEQ ID NO: 1054 / SEQ ID NO: 1079), Blocking Element 23 (SEQ ID NO: 1055 / SEQ ID NO: 1080), Blocking Element 24 (SEQ ID NO: 1056 / SEQ ID NO: 1081), Blocking Element 25 (SEQ ID NO: 1057 / SEQ ID NO: 1082).

47. An inducible IL-18 prodrug comprising an IL-18 polypeptide, a half-life extension element, an IL-18 blocking element and a protease cleavable linker, wherein the IL-18 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-18 prodrug comprises: i. a first polypeptide comprising the IL-18 polypeptide, wherein the IL-18 polypeptide comprises any one of SEQ ID NOs: 2-170, 200-209, or 211-245; a half-life extension element, wherein the half-life extension element comprises any one of SEQ ID NOs: 602-606 and 608-654, or an amino acid sequence of at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 602-606 and 608-654; an antigen binding portion of an antibody heavy chain, wherein the antibody binding portion of the heavy chain comprises any one of SEQ ID NOs: 171, 173, 318, 320, 331, 322, 329, 330, 337, 277 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 339, 353, 354, 655-660, 662, 664, 733-736, 738-742, 820-823, 875-877, 920, 966-969, 1005, 1006, 1010, or 1011. wherein the IL-18 polypeptide and the antigen binding portion of the antibody heavy chain and / or half-life extension element are operably linked by the protease cleavable linker, wherein the protease cleavable linker comprises any one of SEQ ID NOs: 354-379; and ii. a second polypeptide comprising at least an antigen binding portion of an antibody light chain that is complementary to the heavy chain in the first polypeptide and together with said light chain forms an IL-18 binding site, wherein the antibody light chain comprises any one of SEQ ID NOs: 172, 174, 187, 188, 311, 312, 314-316, 319, 321, 323, 815, 816, 862-872, 1012-1016, or 1019.

48. The inducible IL-18 prodrug of claim 47, wherein the IL-18 blocking element is selected: Blocking Element 1 (SEQ ID NO: 1033 / SEQ ID NO: 1058), Blocking Element 2 (SEQ ID NO: 1034 / SEQ ID NO: 1059), Blocking Element 3 (SEQ ID NO: 1035 / SEQ ID NO: 1060), Blocking Element 4 (SEQ ID NO: 1036 / SEQ ID NO: 1061), Blocking Element 5 (SEQ ID NO: 1037 / SEQ ID NO: 1062), Blocking Element 6 (SEQ ID NO: 1038 / SEQ ID NO: 1063), Blocking Element 7 (SEQ ID NO: 1039 / SEQ ID NO: 1064), Blocking Element 8 (SEQ ID NO: 1040 / SEQ ID NO: 1065), Blocking Element 9 (SEQ ID NO: 1041 / SEQ ID NO: 1066), Blocking Element 10 (SEQ ID NO: 1042 / SEQ ID NO: 1067), Blocking Element 11 (SEQ ID NO: 1043 / SEQ ID NO: 1068), Blocking Element 12 (SEQ ID NO: 1044 / SEQ ID NO: 1069), Blocking Element 13 (SEQ ID NO: 1045 / SEQ ID NO: 1070), Blocking Element 14 (SEQ ID NO: 1046 / SEQ ID NO: 1071), Blocking Element 15 (SEQ ID NO: 1047 / SEQ ID NO: 1072), Blocking Element 16 (SEQ ID NO: 1048 / SEQ ID NO: 1073), Blocking Element 17 (SEQ ID NO: 1049 / SEQ ID NO: 1074), Blocking Element 18 (SEQ ID NO: 1050 / SEQ ID NO: 1075), Blocking Element 19 (SEQ ID NO: 1051 / SEQ ID NO: 1076), Blocking Element 20 (SEQ ID NO: 1052 / SEQ ID NO: 1077), Blocking Element 21 (SEQ ID NO: 1053 / SEQ ID NO: 1078), Blocking Element 22 (SEQ ID NO: 1054 / SEQ ID NO: 1079), Blocking Element 23 (SEQ ID NO: 1055 / SEQ ID NO: 1080), Blocking Element 24 (SEQ ID NO: 1056 / SEQ ID NO: 1081), Blocking Element 25 (SEQ ID NO: 1057 / SEQ ID NO: 1082).

49. The inducible IL-18 prodrug of any one of claims 1-30, wherein the inducible IL-18 prodrug comprises Compound 80 (SEQ ID NO: 334 / SEQ ID NO: 312), Compound 87 (SEQ ID NO: 341 / SEQ ID NO: 862), Compound 90 (SEQ ID NO: 665 / SEQ ID NO: 864), Compound 94 (SEQ ID NO: 670 / SEQ ID 278 \\4127-7386-4278 v1Attorney Docket No.: 761146.362320 NO: 867), Compound 101 (SEQ ID NO: 864 / SEQ ID NO: 312), Compound 107 (SEQ ID NO: 690 / SEQ ID NO: 312), Compound 128 (SEQ ID NO: 717 / SEQ ID NO: 871), Compound 129 (SEQ ID NO: 718 / SEQ ID NO: 871), Compound 148 (SEQ ID NO: 747 / SEQ ID NO: 870), Compound 160 (SEQ ID NO: 759 / SEQ ID NO: 188), Compound 178 (SEQ ID NO: 776 / SEQ ID NO: 872), Compound 185 (SEQ ID NO: 783 / SEQ ID NO: 187), Compound 199 (SEQ ID NO: 801 / SEQ ID NO: 187), Compound 206 (SEQ ID NO: 808 / SEQ ID NO: 312), Compound 212 (SEQ ID NO: 840 / SEQ ID NO: 187), Compound 213 (SEQ ID NO: 841 / SEQ ID NO: 187), Compound 223 (SEQ ID NO: 851 / SEQ ID NO: 314), Compound 228 (SEQ ID NO: 921 / SEQ ID NO: 1015), Compound 232 (SEQ ID NO: 927 / SEQ ID NO: 1016), Compound 237 (SEQ ID NO: 942 / SEQ ID NO: 1015), or Compound 244 (SEQ ID NO: 949 / SEQ ID NO: 1015).

50. A inducible IL-18 prodrug of any one of claims 1-31, wherein the inducible IL-18 prodrug can be selected from the group consisting of Compounds 277-279.

51. The inducible IL-18 prodrug of any one of claims 1-30, wherein the inducible IL-18 prodrug comprises Compound 277 (SEQ ID NO: 1101 / SEQ ID NO: 1102), Compound 278 (SEQ ID NO: 1103 / SEQ ID NO: 187), or Compound 279 (SEQ ID NO: 289 / SEQ ID NO: 1104). 279 \\4127-7386-4278 v1

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