Administration of engineered interleukin-18 therapeutics

Abstract

Methods of administering an engineered interleukin-18 (IL-18) polypeptide to a subject are provided. Embodiments of the methods generally involve administering the engineered IL-18 polypeptide via an intravenous route of administration to the subject in order to deliver an effective amount of the engineered IL-18 polypeptide to the subject. In some embodiments, the administered engineered IL-18 polypeptide is a decoy-resistant IL-18 polypeptide. The methods of the present disclosure are useful for administering engineered IL-18 polypeptides to subjects in need of treatment with such agents, including for the treatment of cancer, including solid tumors. Also provided are compositions and kits useful in practicing the herein disclosed methods.

Description

ADMINISTRATION OF ENGINEERED INTERLEUKIN- 18 THERAPEUTICSCROSS-REFERENCE

[0001] This application claims benefit of U.S. Provisional Patent Application No. 63 / 508,694 filed June 16, 2023, which application is incorporated herein by reference in its entirety.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The sequence listing of the present application is submitted electronically on as an ASCII formatted sequence listing with a file name “ST-013-WOl.xml”, creation date of June 9, 2024, and a size of 112kb. This sequence listing submitted is part of the specification and is herein incorporated by reference in its entirety.INTRODUCTION

[0003] According to the National Cancer Institute (NCI) Surveillance, Epidemiology, and End Results Program (SEER), approximately 606,520 people will have died of cancer in the United States (US) in 2020. Despite advances in cancer prevention, diagnosis, and treatment, an estimated 1,806,590 people in the US will have been diagnosed with a new cancer in 2020. Thus, cancer remains a continuing public health concern of major proportions.

[0004] While significant progress has been made in the treatment of many blood cancers, advances in the treatment of many solid cancers have lagged behind those for blood cancers. Solid tumors, including melanoma of the skin, kidney and renal pelvis cancers, squamous cell carcinoma of the head and neck, breast cancers, and lung and bronchitis cancers, collectively account for over 40% of all new cancer cases in the US and were responsible for more than an estimated 200,000 deaths in 2020. Also, despite advancements in treatment, the 5-year relative survival rate for some solid tumors remains low.

[0005] Immune checkpoint inhibitors have made a huge impact in cancer treatment but are associated with tumor shrinkage only in a small proportion of patients, and some tumors are completely refractory to this type of treatment, at least as a single agent.SUMMARY

[0006] Methods of administering an engineered interleukin- 18 (IL-18) polypeptide to a subject are provided. Embodiments of the methods generally involve administering the engineered IL- 18 polypeptide via an intravenous route of administration to the subject in order to deliver an effective amount of the engineered IL- 18 polypeptide to the subject. In some embodiments, the administered engineered IL-18 polypeptide is a decoy-resistant IL-18 polypeptide. The methods of the present disclosure are useful for administering engineered IL-18 polypeptides to subjects in need of treatment with such agents, including for the treatment of cancer, including solidtumors. Also provided are compositions and kits useful in practicing the herein disclosed methods.

[0007] One aspect of the present disclosure provides a method of delivering an effective amount of an engineered IL- 18 polypeptide to a subject in need thereof, the method comprising: administering an amount of the engineered IL-18 polypeptide from 20 pg / kg to 3000 pg / kg to the subject by an intravenous (IV) route of administration, thereby delivering an effective amount of the engineered IL- 18 polypeptide to the subject.

[0008] In some embodiments, the engineered IL-18 polypeptide is a decoy-resistant (DR) IL-18 polypeptide. In some embodiments, the engineered IL- 18 polypeptide comprises an amino acid sequence having at least 85% sequence identity with at least one of SEQ ID NOs: 1, 7, 17-22, 55-57, 62. In some embodiments, the engineered IL-18 polypeptide comprises an amino acid sequence having at least 90% sequence identity with at least one of SEQ ID NOs: 1, 7, 17-22, 55-57, 62. In some embodiments, the engineered IL-18 polypeptide comprises one or more mutations at positions Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DUO, Nl l l, Ml 13, V153, and N155 relative to the wild-type IL-18 amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the engineered IL-18 polypeptide comprises at least 5 mutations relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the at least 5 mutations are at positions selected from Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DUO, Nl l l, Ml 13, V153, and N155. In some embodiments, the engineered IL-18 polypeptide comprises at least 10 mutations relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO:1. In some embodiments, the at least 10 mutations are at positions selected from Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DI 10, N111, Ml 13, V153, and N155. In some embodiments, the engineered IL- 18 polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 7, 17-22, 55-57, 62.

[0009] In some embodiments, the DR IL- 18 polypeptide DR- 18 polypeptide has less than 100% and at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 88%, at least 87%, at least 86%, or at least 85% sequence identity to WT human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO: 102), wherein the X at position (hereinafter “pos ”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K,R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H.

[0010] In some embodiments, the amount of the engineered IL-18 polypeptide administered is from 1 pg / kg to 3000 pg / kg, 5 pg / kg to 3000 pg / kg, 10 pg / kg to 3000 pg / kg, 20 pg / kg to 3000 pg / kg, 30 pg / kg to 3000 pg / kg, 1 pg / kg to 2000 pg / kg, 5 pg / kg to 2000 pg / kg, 10 pg / kg to 2000 pg / kg, 20 pg / kg to 2000 pg / kg, 30 pg / kg to 2000 pg / kg, 1 pg / kg to 1200 pg / kg, 5 pg / kg to 1200 pg / kg, 10 pg / kg to 1200 pg / kg, 20 pg / kg to 1200 pg / kg, 30 pg / kg to 1200 pg / kg, 1 pg / kg to 600 pg / kg, 5 pg / kg to 600 pg / kg, 10 pg / kg to 600 pg / kg, 20 pg / kg to 600 pg / kg, or 30 pg / kg to 600 pg / kg. In some embodiments, the amount is administered as a single IV infusion. In some embodiments, administering the effective amount of the engineered IL- 18 polypeptide comprises achieving a maximum concentration observed (Cmax) of at least 1000 ng / mL of the engineered IL-18 polypeptide in plasma of the subject within 1 day of the administration. In some embodiments, the method comprises achieving a Cmax of at least 1000 ng / mL of the engineered IL-18 polypeptide in plasma of the subject within 12 hours of the administration. In some embodiments, the method comprises achieving a Cmax of at least 1000 ng / mL of the engineered IL- 18 polypeptide in plasma of the subject within 6 hours of the administration.

[0011] In some embodiments, the method comprises repeating the administration weekly, once every two weeks, or once every three weeks. In some embodiments, administration is repeated for at least four weeks. In some embodiments, administration is repeated for at least seven weeks. In some embodiments, the administering results in an engineered IL- 18 polypeptide area under the curve (AUC) accumulation ratio (AR) that is at least five times higher than the engineered IL-18 polypeptide AUC AR resulting from a corresponding subcutaneous route of administration. In some embodiments, the administering results in an engineered IL- 18 polypeptide Cmax AR that is at least five times higher than the engineered IL- 18 polypeptide Cmax AR resulting from a corresponding subcutaneous route of administration.

[0012] In some instances, the amount of an engineered IL- 18 polypeptide that is delivered by an IV route of administration is equivalent to the amount of engineered IL-18 polypeptide administered by a different route of administration, such as e.g., subcutaneously, intramuscularly, intradermally, or the like. In some instances, two amounts delivered by different routes of administration may be equivalent because an equal (i.e., the same) amount isdelivered by both routes of administration. For example, 60 pg / kg delivered intravenously as compared to 60 pg / kg delivered subcutaneously. In some instances, two amounts delivered by different routes of administration may be equivalent because the two amounts result in analogous physiological concentrations of the engineered IL- 18 polypeptide within a subject, including e.g., within the blood or plasma of the subject. For example, 120 pg / kg delivered subcutaneously results in a Cmax that is analogous to the Cmax resulting from 60 pg / kg delivered intravenously. Parameters that may be compared to identify analogous physiological concentrations between two different routes of administration are not necessarily limited to Cmax and may, e.g., include other parameters such as e.g., area under the curve (AUC) parameters, such as e.g., AUCiast, AUCinf , AUCT, AUC Accumulation Ratio (AR), and the like.

[0013] As described elsewhere herein, administration of an engineered IL- 18 polypeptide by an IV ROA has advantages over administration of the engineered IL- 18 polypeptide by other RO As, such as subcutaneous, intramusular, interdermal, and the like. One advantage is superior pharmokinetic parameters, as compared to other delivery methods, facilitating delivery of an amnout of the engineered IL- 18 polypeptide to the subject effective to treat the subject for a condition, such as cancer. One advantage is reduced risks related to injection site reactions (ISRs), such as reduced risk of developing an ISR, reduced risk of developing one or more symptoms of an ISR, reduced risk of developing a Grade 3 ISR, reduced risk of developing a Grade 2 or higher ISR, and the like.

[0014] Risk reduction (or increase) may be quantified be any convenient method including, but not limited to, e.g., a “risk ratio” (i.e., a measure of the risk of a certain event happening in one subject or group of subjects compared to the risk of the same event happening in another subject or group of subjects, sometimes referred to as “relative risk”). A risk ratio of one (1) means there is no difference between two groups in terms of their risk of the event (e.g., occurance of an ISR), whereas a risk ratio of greater than one (>1) or of less than one (<1) usually means that the variable being tested either increases (risk ratio >1) or decreases (risk ratio <1) the risk of the event.

[0015] In some embodiments, methods of the present disclosure reduce the risk of developing an ISR (e.g., as compared to a corresponding method using a ROA other than IV adminstration) by a risk ratio of 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, or 0.1 or less. In some embodiments, methods of the present disclosure reduce the risk of developing one or more symtoms of an ISR (e.g., ulcers, necrosis, tissue damage, pain, lipodystrophy, edema, phlebitis, tenderness, erythema, and itching) by a risk ratio of 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, or 0.1 or less. In some embodiments, methods of the present disclosure reduce the risk of developing an a Grade3 (e.g., according to CTCAEv5) ISR by a risk ratio of 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, or 0.1 or less. In some embodiments, methods of the present disclosure reduce the risk of developing an a Grade 2 (e.g., according to CTCAEv5) or higher ISR by a risk ratio of 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, or 0.1 or less. In some instances, such reduction in risk may be expressed by other means, such as e.g., absolute risk, odds ratio, and the like. Comparison between two different methods, to assess the relative risk, will generally involve comparison of two equivalent amounts of administered engineered IL- 18 polypeptide, including e.g., where the amounts administered are equivalent in that equal amounts of are administered, in that the amounts administered results in analogous physilogical concentrations within the subject, or the like. In some instances, the methods of the present disclosure result in less Grade 3 ISRs, less Grade 2 or higher ISRs, fewer ISR symptoms, less severe ISR symptoms, and / or less ISRs, e.g., as compared to methods employing delivery methods other than IV, such as e.g., subcutaneous injection.

[0016] In some embodiments, the method further comprising pre-treating the subject with an effective amount of a B-cell depleter prior to administering the amount of the engineered IL- 18 polypeptide, optionally wherein the B-cell depleter comprises an anti-CD20 antibody, optionally wherein the anti-CD20 antibody is obinutuzumab.

[0017] One aspect of the disclosure provides a method of administering an engineered interleukin 18 (IL-18) polypeptide to a subject in need thereof, the method comprising: pretreating the subject with an effective amount of a B-cell depleter to deplete B-cells in the subject; and administering the engineered IL- 18 polypeptide to the B-cell depleted subject.

[0018] In some embodiments, the engineered IL-18 polypeptide is a decoy-resistant (DR) IL-18 polypeptide. In some embodiments, the B-cell depleter is a B-cell depleting antibody. In some embodiments, the antibody is an anti-CD20 antibody. In some embodiments, the B-cell depleting antibody is obinutuzumab. In some embodiments, the engineered IL-18 polypeptide or DR IL-18 polypeptide is administered by an IV ROA, including e.g., at a dose of at least 1 pg / kg, at least 5 pg / kg, at least 10 pg / kg, at least 20 pg / kg, at least 30 pg / kg, at least 60 pg / kg, at least 120 pg / kg, or at least 240 pg / kg.

[0019] In some embodiments, of any of the herein described methods, the subject has a cancer and the method comprises administering the engineered IL-18 polypeptide to treat the subject for the cancer. In some embodiments, the cancer comprises a solid tumor. In some embodiments, the cancer is selected from the group consisting of melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-hightumors, high tumor mutation burden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer. In some embodiments, the cancer has lost surface expression of MHC class I. In some embodiments, the cancer is checkpoint inhibitor resistant. In some embodiments, the cancer is an anti-PDl -resistant tumor.BRIEF DESCRIPTION OF THE FIGURES

[0020] FIG. 1 depicts plasma concentrations of a test compound in individual non-human primates over the first six hours of Day 1 following a single 1 mg / kg dose of the test compound by either subcutaneous injection (SC) or intravenous infusion (IV).DEFINITIONS

[0021] The terms “subject,” “individual,” or “patient” are often used interchangeably herein. A “subject” comprises a biological entity containing expressed genetic materials. In some embodiments, the subject comprises an animal, mammal, primate, or human. In some embodiments, the subject is diagnosed with cancer. In some embodiments, the subject has a solid tumor. In some embodiments, the subject has a liquid tumor.

[0022] The term “subject in need thereof’ as used herein refers to a subject diagnosed with or suspected of having cancer as defined herein.

[0023] As used herein, the terms “treat,” “treated,” “treating,” “treatment,” and the like are used in reference to an intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to prevention or amelioration of symptoms or of an underlying disorder being treated. Also, a therapeutic benefit can be achieved with prevention or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or a condition, delaying or eliminating the onset of symptoms of a disease or a condition, slowing, halting, or reversing the progression of a disease or a condition, or any combination thereof. For prophylactic benefit, a subject at risk of developing a particular disease or a condition, or to a subject reporting one or more of the physiological symptoms of a disease or a condition may undergo treatment.

[0024] The term “prevent”, “preventing”, “prevention” and their grammatical equivalents as used herein, means avoiding or delaying the onset of symptoms associated with a disease or condition in a subject that has not developed such symptoms at the time the administering of an agent or compound commences.

[0025] The term “therapeutic effect” refers to some extent of relief of one or more of the symptoms of a disorder (e.g., a neoplasia, tumor, or infection by an infectious agent or an autoimmune disease) or its associated pathology. “Therapeutically effective amount” as used herein refers to an amount of an agent which is effective, upon single or multiple dose administration to the cell or subject, in prolonging the survivability of the patient with such a disorder, reducing one or more signs or symptoms of the disorder, preventing or delaying, and the like beyond that expected in the absence of such treatment. “Therapeutically effective amount” is intended to qualify the amount required to achieve a therapeutic effect. A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the “therapeutically effective amount” (e.g., ED50) of the pharmaceutical composition required.

[0026] The terms “neoplasia” and “cancer” refers to any disease that is caused by or results in inappropriately high levels of cell division, inappropriately low levels of apoptosis, or both. Merkel cell carcinoma is one non-limiting example of a cancer. The terms “cancer” or “tumor” or “hyperproliferative disorder” refer to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. Cancer cells are often in the form of a tumor, but such cells can exist alone within an animal, or can be a nontumorigenic cancer cell, such as a leukemia cell.

[0027] The term “liquid tumor” refers to cancers occurring, or the presence of cancerous cells, in bodily fluids. Accordingly, liquid tumors are cancers that affect the blood, bone marrow, and / or lymphatic systems. Liquid tumors include hematological (or “heme” or blood) cancers, such as e.g., a leukemia, a lymphoma, a myelodysplastic syndrome, a myeloproliferative disorder, and myeloma. The most common types of hematological cancers include leukemia, lymphoma, and myeloma. By contrast, a “solid tumor” refers to a grouping of cancerous cells that forms a solid mass in a tissue of a subject, such as bone tissue, muscle tissue, epithelial tissue, organ tissue (e.g., liver, lung, stomach, colon, kidney, or brain tissue), and the like. Types of solid tumors include e.g., carcinomas, sarcomas, lymphomas, melanomas, neuroendocrine tumors, germ cell tumors, gliomas, and the like.

[0028] An “agent” can include any type of molecule and includes, but is not limited to, an antibody, a peptide, a protein, a polynucleotide (e.g., an oligonucleotide, RNA, or DNA), a small molecule, derivatives thereof and analogs thereof.

[0029] The terms “peptide”, “polypeptide” and “protein” are used herein interchangeably to describe a series of at least two amino acids covalently linked by peptide bonds or modified peptide bonds such as isosteres. No limitation is placed on the maximum number of amino acids which may comprise a peptide or protein. Furthermore, the term polypeptide extends tofragments, analogues and derivatives of a peptide, wherein said fragment, analogue or derivative retains the same biological functional activity as the peptide from which the fragment, derivative or analogue is derived. A polypeptide as used herein may be encoded by a recombinant nucleic acid.

[0030] A “fragment” is a portion of a protein or nucleic acid that is substantially identical to a reference protein or nucleic acid. In some embodiments, the portion retains at least 50%, 75%, or 80%, or 90%, 95%, or even 99% of the biological activity of the reference protein or nucleic acid described herein.

[0031] A "functional derivative" or “functional fragment” of a native sequence polypeptide (e.g., antibody) is a compound having a qualitative biological property in common with a native sequence polypeptide. Functional derivatives or fragments include, but are not limited to, fragments of a native sequence and derivatives of a native sequence polypeptide and its fragments, provided that they have a biological activity in common with a corresponding native sequence polypeptide. The terms "derivative" and “fragment” encompass both amino acid sequence variants of polypeptide and covalent modifications thereof.

[0032] As used herein, the term “antibody” refers to any form of immunoglobulin molecule that exhibits the desired biological or binding activity. Thus, it is used in the broadest sense and specifically covers, but is not limited to, monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies e.g., bispecific antibodies), humanized, fully human antibodies, and chimeric antibodies, and may include post- translational modifications thereof (e.g., C-terminal Lysine clipping in the heavy chain, conversion of glutamine or glutamic acid to pyroglutamate) that may occur when an antibody is recombinantly expressed in host cells (e.g., CHO cells), or during purification / storage. “Parental antibodies” are antibodies obtained by exposure of an immune system to an antigen prior to modification of the antibodies for an intended use, such as humanization of an antibody for use as a human therapeutic. As used herein, the term “antibody” encompasses not only intact polyclonal or monoclonal antibodies, but also, unless otherwise specified, fusion proteins comprising an antigen binding fragment thereof that competes with the intact antibody for specific.

[0033] In general, the basic antibody structural unit comprises a tetramer. Each tetramer includes two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The variable regions of each light / heavy chain pair form the antibody binding site. Thus, in general, an intact antibody has two binding sites. The carboxy -terminal portion of theheavy chain may define a constant region primarily responsible for effector function. Typically, human light chains are classified as kappa and lambda light chains. Furthermore, human heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon, and define the antibody’s isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a “J” region of about 12 or more amino acids, with the heavy chain also including a “D” region of about 10 more amino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, N.Y. (1989).

[0034] “Variable regions” or “V region” or “V chain” as used herein means the segment of IgG chains which is variable in sequence between different antibodies. A “variable region” of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. The variable region of the heavy chain may be referred to as “VH ” The variable region of the light chain may be referred to as “VL.”

[0035] Typically, the variable regions of both the heavy and light chains comprise three hypervariable regions, also called complementarity determining regions (CDRs), which are located within relatively conserved framework regions (FR). The CDRs are usually aligned by the framework regions, enabling binding to a specific epitope. In general, from N-terminal to C- terminal, both light and heavy chains variable domains comprise FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. As referred to herein the light chain CDRs are CDRL1, CDRL2 and CDRL3, respectively, and the heavy chain CDRs are CDRH1, CDRH2 and CDRH3, respectively. The assignment of amino acids to each domain is, generally, in accordance with the definitions of Sequences of Proteins of Immunological Interest, Kabat, et al. National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ. No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32: 1-75; Kabat, et al., (1977) J. Biol. Chem. 252:6609-6616; Chothia, et al., (1987) J Mol. Biol. 196:901- 917 or Chothia, et al., (1989) Nature 342:878-883.

[0036] A “CDR” refers to one of three hypervariable regions (Hl, H2, or H3) within the nonframework region of the antibody VH P-sheet framework, or one of three hypervariable regions (LI, L2, or L3) within the non-framework region of the antibody VL P-sheet framework. Accordingly, CDRs are variable region sequences interspersed within the framework region sequences. CDR regions are well known to those skilled in the art and have been defined by, for example, Kabat as the regions of most hypervariability within the antibody variable domains. CDR region sequences also have been defined structurally by Chothia as those residues that are not part of the conserved P -sheet framework, and thus are able to adapt to different conformations. Both terminologies are well recognized in the art. CDR region sequences have also been defined by AbM, Contact, and IMGT. The positions of CDRs within a canonical antibody variable region have been determined by comparison of numerous structures (Al-Lazikani et aL, 1997, J. Mol. Biol. 273:927-48; Morea et aL, 2000, Methods 20:267-79). Because the number of residues within a hypervariable region varies in different antibodies, additional residues relative to the canonical positions are conventionally numbered with a, b, c and so forth next to the residue number in the canonical variable region numbering scheme (Al- Lazikani et cd.. supra). Such nomenclature is similarly well known to those skilled in the art. Correspondence between the numbering system, including, for example, the Kabat numbering and the IMGT unique numbering system, is well known to one skilled in the art and shown below in Table 1. In some embodiments, the CDRs are as defined by the Kabat numbering system. In other embodiments, the CDRs are as defined by the IMGT numbering system. In yet other embodiments, the CDRs are as defined by the AbM numbering system. In still other embodiments, the CDRs are as defined by the Chothia numbering system. In yet other embodiments, the CDRs are as defined by the Contact numbering system.Table 1. Correspondence between the CDR Numbering Systems

[0037] Chimeric antibody” refers to an antibody in which a portion of the heavy and / or light chain contains sequences derived from a particular species (e.g., human) or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is derived from another species (e.g., mouse) or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity.

[0038] “Human antibody” refers to an antibody that comprises human immunoglobulin protein sequences or derivatives thereof. A human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, “mouse antibody” or “rat antibody” refer to an antibody that comprises only mouse or rat immunoglobulin sequences or derivatives thereof, respectively.

[0039] “Humanized antibody” refers to forms of antibodies that contain sequences from nonhuman (e.g., murine) antibodies as well as human antibodies. Such antibodies contain minimal sequence derived from non-human immunoglobulin. In general, the humanized antibody willcomprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. The prefix “hum”, “hu” or “h” may be added to antibody clone designations when necessary to distinguish humanized antibodies from parental rodent antibodies. The humanized forms of rodent antibodies will generally comprise the same CDR sequences of the parental rodent antibodies, although certain amino acid substitutions may be included to increase affinity, increase stability of the humanized antibody, or for other reasons.

[0040] “Monoclonal antibody” or “mAb” or “Mab”, as used herein, refers to a population of substantially homogeneous antibodies, z.e., the antibody molecules comprising the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include a multitude of different antibodies having different amino acid sequences in their variable domains, particularly their CDRs, which are often specific for different epitopes. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present disclosure may be made by the hybridoma method first described by Kohler et al. (1975) Nature 256: 495, or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597, for example. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0041] As used herein, unless otherwise indicated, “antibody fragment” or “antigen binding fragment” refers to a fragment of an antibody that retains the ability to bind specifically to the antigen, e.g., fragments that retain one or more CDR regions and the ability to bind specifically to the antigen. An antibody that “specifically binds to” CD20 is an antibody that exhibits preferential binding to CD20 (as appropriate) as compared to other proteins, but this specificity does not require absolute binding specificity. An antibody is considered “specific” for its intended target if its binding is determinative of the presence of the target protein in a sample, e.g., without producing undesired results such as false positives. Antibodies, or binding fragments thereof, will bind to the target protein with an affinity that is at least two-fold greater,preferably at least ten times greater, more preferably at least 20-times greater, and most preferably at least 100-times greater than the affinity with non-target proteins.

[0042] Antigen binding portions include, for example, Fab, Fab’, F(ab’)2, Fd, Fv, fragments including CDRs, and single chain variable fragment antibodies (scFv), and polypeptides that contain at least a portion of an immunoglobulin that is sufficient to confer specific antigen binding to the antigen (e.g., CD20). An antibody includes an antibody of any class, such as IgG, IgA, or IgM (or sub-class thereof), and the antibody need not be of any particular class. Depending on the antibody amino acid sequence of the constant region of its heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The heavy-chain constant regions that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0043] An "antigen" is a structure to which an antibody can selectively bind. A target antigen may be a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. In some embodiments, the target antigen is a polypeptide. In certain embodiments, an antigen is associated with a cell, for example, is present on or in a cell, for example, a cancer cell.

[0044] An "intact" antibody is one comprising an antigen-binding site as well as a constant domain (CL) and at least heavy chain constant regions, CHI, CH2 and CH3. The constant regions may include human constant regions or amino acid sequence variants thereof. In certain embodiments, an intact antibody has one or more effector functions.

[0045] As used herein, the term “immune response” relates to any one or more of the following: specific immune response, non-specific immune response, both specific and non-specific response, innate response, primary immune response, adaptive immunity, secondary immune response, memory immune response, immune cell activation, immune cell-proliferation, immune cell differentiation, and cytokine expression.

[0046] The term “enteral route” of administration refers to the administration via any part of the gastrointestinal tract. Examples of enteral routes include oral, mucosal, buccal, and rectal route, or intragastric route. “Parenteral route” of administration refers to a route of administration other than enteral route. Examples of parenteral routes of administration include intravenous, intramuscular, intradermal, intraperitoneal, intratumor, intravesical, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, transtracheal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal, subcutaneous, or topical administration. Therapeutic agentsand compositions can be administered using any suitable method, such as by oral ingestion, nasogastric tube, gastrostomy tube, injection, infusion, implantable infusion pump, and osmotic pump. A suitable route and method of administration can vary depending on a number of factors such as the specific therapeutic agent being used, the rate of absorption desired, specific formulation or dosage form used, type or severity of the disorder being treated, the specific site of action, and conditions of the patient, and can be readily selected by a person skilled in the art.

[0047] “Chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Classes of chemotherapeutic agents include, but are not limited to: alkylating agents, antimetabolites, kinase inhibitors, spindle poison plant alkaloids, cytoxic / antitumor antibiotics, topoisomerase inhibitors, photosensitizers, anti -estrogens and selective estrogen receptor modulators (SERMs), anti-progesterones, estrogen receptor down-regulators (ERDs), estrogen receptor antagonists, leutinizing hormone-releasing hormone agonists, anti -androgens, aromatase inhibitors, EGFR inhibitors, VEGF inhibitors, and anti-sense oligonucleotides that inhibit expression of genes implicated in abnormal cell proliferation or tumor growth. Chemotherapeutic agents useful in the treatment methods of the invention include cytostatic and / or cytotoxic agents.

[0048] “Conservatively modified variants” or “conservative substitution” refers to substitutions of amino acids in a protein with other amino acids having similar characteristics (e.g., charge, sidechain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity, etc.), such that the changes can frequently be made without altering the biological activity or other desired property of the protein, such as antigen affinity and / or specificity. Those of skill in this art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th Ed.)). In addition, substitutions of structurally or functionally similar amino acids are less likely to disrupt biological activity. Exemplary conservative substitutions include: for Ala (A): Gly, Ser; for Arg (R): Lys, His; for Asn (N): Gin, His; for Asp (D): Glu, Asn; for Cys (C): Ser, Ala; for Gin (Q): Asn; for Glu (E): Asp, Gin; for Gly (G): Ala; for His (H): Asn, Gin; for He (I): Leu, Vai; for Leu (L): He, Vai; for Lys (K): Arg, His; for Met (M): Leu, lie, Tyr; for Phe (F): Tyr, Met, Leu; for Pro (P): Ala; for Ser (S): Thr; for Thr (T): Ser; for Trp (W): Tyr, Phe; for Tyr (Y): Trp, Phe; and for Vai (V): He, Leu.

[0049] “RECIST 1.1 Response Criteria”, and related terms, as used herein means the definitions set forth in Eisenhauer, E.A. et al., Eur. J. Cancer 45:228-247 (2009) for target lesions or nontarget lesions, as appropriate based on the context in which response is being measured.

[0050] Sustained response” means a sustained therapeutic effect after cessation of treatment as described herein. In some embodiments, the sustained response has a duration that is at least the same as the treatment duration, or at least 1.5, 2.0, 2.5 or 3 times longer than the treatment duration.

[0051] “Non-responder patient”, when referring to a specific anti-tumor response to a treatment described herein, means the patient did not exhibit an anti-tumor response.

[0052] “Responder patient” when referring to a specific anti-tumor response to a treatment described herein, means the patient exhibited an anti-tumor response.

[0053] Treat” or “treating” cancer as used herein means to administer an agent to a subject having cancer or diagnosed with cancer to achieve at least one positive therapeutic effect, such as, for example, reduced number of cancer cells, reduced tumor size, reduced rate of cancer cell infiltration into peripheral organs, or reduced rate of tumor metastasis or tumor growth, comprising administration by parenteral or enteral routes, including e.g., oral, mucosal, intradermal, intravenous, subcutaneous, intramuscular delivery, and / or any other methods of physical delivery described herein or known in the art. Typically, the agent(s) of the treatment method are administered in an amount effective to alleviate one or more disease symptoms in the treated subject or population, whether by inducing the regression of or inhibiting the progression of such symptom(s) by any clinically measurable degree. The amount of the agent(s) of the treatment method that is effective to alleviate any particular disease symptom can vary according to factors such as the disease state, age, and weight of the patient, and the ability of the therapeutic combination to elicit a desired response in the subject. Whether a disease symptom has been alleviated can be assessed by any clinical measurement typically used by physicians or other skilled healthcare providers to assess the severity or progression status of that symptom. “Treatment” can include one or more of the following: inducing / increasing an antitumor immune response, decreasing the number of one or more tumor markers, halting or delaying the growth of a tumor or blood cancer or progression of disease such as cancer, stabilization of disease, inhibiting the growth or survival of tumor cells, eliminating or reducing the size of one or more cancerous lesions or tumors, decreasing the level of one or more tumor markers, ameliorating or abrogating the clinical manifestations of disease, reducing the severity or duration of the clinical symptoms, prolonging the survival or patient relative to the expected survival in a similar untreated patient, and inducing complete or partial remission of a cancerous condition, wherein the disease is cancer, more specifically e.g., a solid tumor.

[0054] The amount of a therapeutic agent that is effective to alleviate any particular disease symptom can vary according to factors such as the disease state, age, and weight of the patient, and the ability of the drug to elicit a desired response in the subject. Whether a disease symptomhas been alleviated can be assessed by any clinical measurement typically used by physicians or other skilled healthcare providers to assess the severity or progression status of that symptom.

[0055] Positive therapeutic effects in cancer can be measured in a number of ways (See, W. A. Weber, J. NucL Med. 50: 1 S-10S (2009)). For example, with respect to tumor growth inhibition, according to NCI standards, a T / C = 42% is the minimum level of anti-tumor activity. A T / C < 10% is considered a high anti -tumor activity level, with T / C (%) = Median tumor volume of the treated / Median tumor volume of the control x 100. In some embodiments, the treatment achieved by a therapy of the disclosure is any of partial response (PR), complete response (CR), objective response (OR), progression-free survival (PFS), disease-free survival (DFS), and overall survival (OS). PFS, also referred to as “Time to Tumor Progression” indicates the length of time during and after treatment that the cancer does not grow, and includes the amount of time patients have experienced a CR or PR, as well as the amount of time patients have experienced SD. DFS refers to the length of time during and after treatment that the patient remains free of disease. OS refers to a prolongation in life expectancy as compared to naive or untreated individuals or patients. In some embodiments, response to a therapy of the disclosure is any of PR, CR, PFS, DFS, or OR that is assessed using RECIST 1.1 response criteria. The treatment regimen for a therapy of the disclosure that is effective to treat a cancer patient can vary according to factors such as the disease state, age, and weight of the patient, and the ability of the therapy to elicit an anti-cancer response in the subject. While an embodiment of any of the aspects of the disclosure may not be effective in achieving a positive therapeutic effect in every subject, it should do so in a statistically significant number of subjects as determined by any statistical test known in the art such as the Student’s t-test, the chi2-test, the U-test according to Mann and Whitney, the Kruskal-Wallis test (H-test), Jonckheere-Terpstra-test and the Wilcoxon test. In some instances, e.g., relating to randomized oncology studies, other tests may be employed, including e.g., log-rank testing, hazard ratios, and the like.

[0056] A "biological sample" encompasses a variety of sample types obtained from an individual or a population of individuals and can be used in a diagnostic, monitoring or screening assay. The definition encompasses blood and other liquid samples of biological origin, solid tissue samples such as a biopsy specimen or tissue cultures or cells derived therefrom and the progeny thereof. The definition also includes samples that have been manipulated in any way after their procurement, such as by mixing or pooling of individual samples, treatment with reagents, solubilization, or enrichment for certain components, such as cells, polynucleotides, polypeptides, etc. The term "biological sample" encompasses a clinical sample, and also includes cells in culture, cell supernatants, cell lysates, serum, plasma, biological fluid, and tissue samples. The term "biological sample" includes urine, saliva, cerebrospinal fluid,interstitial fluid, ocular fluid, synovial fluid, blood fractions such as plasma and serum, and the like. The term "biological sample" also includes solid tissue samples, tissue culture samples, and cellular samples.

[0057] The term “unit dosage form,” as used herein, refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a subject active agent calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for a subject active agent depend on the particular compound employed and the effect to be achieved, and the pharmacodynamics associated with each compound in the host.

[0058] “Cytokine Release Syndrome” or CRS, as used herein, refers to an acute systemic inflammatory syndrome that can be triggered by a variety of factors such as infections and may occur after treatment with some types of immunotherapy, such as monoclonal antibodies and chimeric antigen receptor (CAR) T cell therapies, as well as some non-protein-based cancer drugs. CRS is characterized by a large, rapid increase in cytokines and inflammatory response. Signs and symptoms of CRS include fever, fatigue, nausea, headache, rash, arthralgia, myalgia, rapid heartbeat, hypotension, and trouble breathing. CRS can progress to an uncontrolled systemic inflammatory response with vasopressor-requiring circulatory shock, vascular leakage, disseminated intravascular coagulation, and multi-organ system failure. Patients may have a mild reaction, or reactions may be severe or life threatening. Grading of CRS can be performed according to the Consensus American Society for Transplantation and Cellular Therapy (ASTCT) grading system, such as e.g., that described in Lee et al. Biology of Blood andMarrow Transplantation. 25 (2019) 625 - 639; the disclosure of which is incorporated herein by reference in its entirety.DETAILED DESCRIPTION

[0059] Methods of administering an engineered interleukin- 18 (IL-18) polypeptide to a subject are provided. Embodiments of the methods generally involve administering the engineered IL- 18 polypeptide via an intravenous route of administration to the subject in order to deliver an effective amount of the engineered IL- 18 polypeptide to the subject. In some embodiments, the administered engineered IL-18 polypeptide is a decoy -resistant IL-18 polypeptide. The methods of the present disclosure are useful for administering engineered IL-18 polypeptides to subjects in need of treatment with such agents, including for the treatment of cancer, including solid tumors. Also provided are compositions and kits useful in practicing the herein disclosed methods.

[0060] Before the present invention is described in greater detail, it is to be understood that this invention is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0061] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges can independently be included in the smaller ranges and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0062] Certain ranges are presented herein with numerical values being preceded by the term "about." The term "about" is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating unrecited number can be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number. The term “about” or “approximately” can when referring to a measurable value such as a parameter, an amount, a temporal duration, and the like, encompass variations of + / -20% or less, + / -10% or less, + / -5% or less, or + / -1% or less of and from the specified value, insofar such variations are appropriate to perform in the present disclosure.

[0063] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, representative illustrative methods and materials are now described.

[0064] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of priorinvention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0065] It is noted that, as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.

[0066] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0067] While the apparatus and method has or will be described for the sake of grammatical fluidity with functional explanations, it is to be expressly understood that the claims, unless expressly formulated under 35 U.S.C. §112, are not to be construed as necessarily limited in any way by the construction of "means" or "steps" limitations, but are to be accorded the full scope of the meaning and equivalents of the definition provided by the claims under the judicial doctrine of equivalents, and in the case where the claims are expressly formulated under 35 U.S.C. §112 are to be accorded full statutory equivalents under 35 U.S.C. §112.METHODS

[0068] The present disclosure includes methods of administering an engineered IL- 18 polypeptide to a subject in need thereof. As described in more detail below, methods of the present disclosure find use in treating a variety of conditions in various subjects, including but not limited to the treatment of cancer.

[0069] IL-18 is a pro-inflammatory cytokine that can stimulate T cells, NK cells, and myeloid cells. IL- 18 has been proposed as an immunotherapeutic agent for the treatment of cancer, given its ability to stimulate anti -tumor cells. IL- 18 binds, and signals through, IL- 18 receptor (IL- 18R; also referred to as Interleukin- 18 receptor 1, IL18R1, IL-18R-1, IL-18R1, IL1 receptor- related protein, IL1RRP, Interleukin- 18 receptor alpha, IL-18Ra, and CD218a). Binding of IL- 18 to IL-18Ra allows for formation of the IL-18 / IL-18Ra / IL-18Rp complex which results in activation of IL-18 signaling and simulation of immune cells. IL-18 binding to IL-18R, and thus signaling through the receptor, may be blocked by IL- 18 binding protein (IL-18BP), which has limited the clinical efficacy of recombinant human wild-type (WT) IL- 18.

[0070] Engineered IL-18 polypeptides, defined as an IL-18 polypeptide having at least one mutated (i.e., artificially changed) amino acid residue as compared to the corresponding WT sequence, are under various stages of development. Exemplary engineered IL-18 polypeptides (also referred to, in some instances, as IL-18 “variants” or simply “engineered IL-18”) include those having at least one mutated residue relative to the WT human IL-18 amino acid sequence of SEQ ID NO: 1. Engineered IL- 18 polypeptides can be engineered to maintain, enhance, reduce, and / or abrogate one or more functions or activities of a WT IL-18 polypeptide, or add an additional function or activity not present in a WT IL-18 polypeptide. Any activity present in a WT IL- 18 polypeptide can be maintained, enhanced, reduced, or abrogated in an engineered IL- 18 polypeptide. Additional functions or activities that can be added to a WT IL- 18 polypeptide in an engineered IL- 18 polypeptide include essentially any protein function not normally or naturally present in the WT IL-18 polypeptide, e.g., by mutation of the WT IL-18 sequence and fusing (e.g., terminal (e.g., N- or C-terminal) fusions or chemically conjugating (e.g., N- or C- terminal conjugation or conjugation to a non-terminal amino acid) additional amino acid sequence (that provides an added function) to the engineered IL- 18 polypeptide. In some instances, useful engineered IL- 18 polypeptides will not include any additional fused amino acid sequence and can consist essentially of the WT IL- 18 amino acid sequence with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) mutated residues. WT human IL-18, after processing of the inactive precursor, is 157 amino acids in length. An engineered IL- 18 polypeptide may be 157 amino acids in length, or may be engineered to be shorter (e.g., by truncation or deletion of one or more amino acids) or longer (e.g., by fusion or conjugation) than 157 amino acids.

[0071] A WT IL-18 polypeptide can be engineered (e.g., by mutation, including e.g., substitution and / or deletion mutation) to reduce (including partially reduce or completely abrogate) its binding to IL-18BP while maintaining its binding to IL-18R and formation of the IL- 18 signaling complex. Such engineered IL- 18 polypeptides, having reduced binding, or no binding, to IL-BP while maintaining binding to IL-18R, and signaling through the receptor, are referred to as decoy-resistant (DR) IL-18 polypeptides. Put another way, DR IL-18 polypeptides are IL-18R agonists that are resistant to inhibition by IL-18BP.

[0072] Useful DR IL-18 polypeptides include, but are not limited to, e.g., those described in PCT Pub. No. W02019 / 051015A1 and US Pat. Pub. No. US20210015891A1, the disclosures of which are incorporated herein by reference in their entirety. As described in more detail below, DR IL-18 polypeptides are useful in medical treatment. In some instances, a decoy -to-the-decoy (D2D) engineered IL-18 polypeptide, as described in more detail below, can be employed, alone or in combination with other agents.

[0073] Useful engineered IL-18 polypeptides, including DR IL-18 polypeptides, also include those which have been stabilized through the addition of one or more stabilizing mutations, e.g., as described in PCT Pub. No. WO2022 / 094473A1, the disclosure of which is incorporated herein by reference in its entirety. As such, polypeptides useful in the herein described methods include stabilized engineered IL-18 polypeptides, including stabilized DR IL-18 polypeptides and stabilized D2D IL- 18 polypeptides, and the like. In some instances, polypeptides useful in the herein described methods include engineered IL- 18 polypeptides, including DR IL- 18 polypeptides and D2D IL- 18 polypeptides, having one or more mutations of cysteine residues present in human WT IL- 18, such as e.g., mutations at C38, C68, both C38 and C68, or the like.

[0074] Engineered IL- 18 polypeptides, including DR IL- 18 polypeptides and D2D IL- 18 polypeptides (as well as stabilized and / or cysteine-mutated versions thereof), useful in the methods of the present disclosure are described in more detail below, and include e.g., engineered IL-18 polypeptides comprising amino acid sequences of any one of SEQ ID NOs:2- 97 and, in some instances, engineered IL-18 polypeptides comprising DR-IL-18 polypeptide amino acid sequences of any one of SEQ ID NOs: 7, 17-22, 55-57, 62 and 63. Dosage amounts, frequency, and other factors and considerations related to the administration of engineered IL-18 polypeptides are described in more detail below.

[0075] As described in the Examples below, subjects treated with an engineered IL- 18 polypeptide by IV ROA experience various advantages, including e.g., advantages as compared to subjects treated with the engineered IL-18 polypeptide by a ROA other than IV, such as e.g., subcutaneous injection (SC). For example, in some instances, subjects experience less or fewer injection-site reactions, e.g., as compared to those treated with an engineered IL- 18 polypeptide by SC ROA (either in a concurrent study arm or from reference data). Fewer injection site reactions (ISRs) may be seen within a population of subjects administered an engineered IL- 18 polypeptide by IV as compared to a comparable population of subjects administered the engineered IL-18 by another route, such as e.g., SC. Less ISRs may be seen in a single subject over a course of therapy that includes multiple doses of an engineered IL- 18 delivered by IV, e.g., as compared to a comparable subject administered a comparable course of therapy delivered by SC. In some instances, both fewer subjects experiencing ISRs and less ISRs per subject are observed. In some instances, fewer Grade 3, including no Grade 3, ISRs are observed in a subject or subjects administered an engineered IL-18 polypeptide, e.g., as compared to a comparable subject or comparable subjects administered the engineered IL- 18 by SC. In some instances, fewer Grade 2, including no Grade 2, ISRs are observed in a subject or population of subjects administered an engineered IL-18 polypeptide, e.g., as compared to a comparable subject or comparable subjects administered the engineered IL-18 by SC. In some instances,ISRs are less severe, e.g., fewer Grade 3 ISRs are observed, fewer Grade 2 ISRs are observed, etc.

[0076] In some instances, a subject, or a population of subjects, experience fewer symptoms or less severe symptoms of ISRs when administered an engineered IL- 18 polypeptide by IV, e.g., as compared to SC, including e.g., fewer instances of or less severe ulcers / ulcerations, necrosis, tissue damage, pain, lipodystrophy, edema, phlebitis, tenderness, erythema, and / or itching. In some instances, subjects treated with an engineered IL-18 polypeptide by IV ROA experience less injection site ulcers and / or increased patient comfort and safety. In some instances, improvements in patient comfort, such as e.g., reduction and / or avoidance of ISRs, can lead to better and / or longer adherence to a treatment protocol and, accordingly, better therapeutic outcomes.

[0077] ISRs resulting from injection of biologies can be severe and require treatment, including e.g., surgical intervention. In some instances, subjects treated according to the methods described herein are at reduced risk of needing treatment, such as e.g., surgical intervention, for an injection site reaction. In some instances, the methods of the present disclosure reduce the need for treatment, such as e.g., surgical intervention, in patients treated with an engineered IL- 18 polypeptide.

[0078] In some instances, subjects experience a combination of advantages, including e.g., improved safety and comfort through a reduction in injection-site reactions and also advantageous pharmacokinetic (PK) profiles (e.g., as compared to an SC ROA). Other combinations of advantages may include, e.g., improved safety and comfort through a reduction in injection-site reactions and also improved therapeutic effect or outcome, such as e.g., an increase in the frequency or duration of a PR, CR, OR, PFS, DFS, and / or OS. Other combinations of advantages may include, e.g., reduced risk of an ISR, and / or reduced need for ISR treatment, such as e.g., surgical intervention, in combination with an improved PK profile of the administered engineered IL-18 polypeptide. Other combinations of advantages may include, e.g., reduced risk of an ISR, and / or reduced need for ISR treatment, such as e.g., surgical intervention, in combination with an improved therapeutic effect or outcome as compared to another ROA (such as SC), including e.g., an increase in the frequency or duration of a PR, CR, OR, PFS, DFS, and / or OS.

[0079] Methods of the present disclosure provide numerous advantages, including but not limited to e.g., improved measures of anti-tumor activity, improved treatment biomarkers, various improved pharmacokinetic parameters with regards to the administered engineered IL- 18 polypeptides, improved treatment with engineered IL-18 polypeptides, improved efficacywith engineered IL- 18 polypeptides, and the like. Advantages of the herein described methods are not limited to those specifically described.

[0080] In some instances, the subject receiving IV administration of an engineered IL- 18 is a naive subject. As used herein, a naive subject refers to a subject that has not previously been treated for their condition, e.g., cancer, tumor, etc. Accordingly, a naive subject having a cancer will have not previously been administered a cancer therapeutic to treat the cancer, such as e.g., chemotherapy, a checkpoint inhibitor, an engineered IL- 18 polypeptide, etc. In some instances, a subject can be “non-naive”, meaning the subject has previously received or undergone treatment for their condition, e.g., having been administered a therapeutic to treat a cancer or tumor. The term “engineered IL- 18 naive” refers to subjects that have not previously received treatment with an engineered IL-18 polypeptide, which includes e.g., subjects that are naive (i.e., they have not previously received any treatment for their condition) and subjects that are non-naive (i.e., they have previously received treatment for their condition, but not treatment with an engineered IL- 18).

[0081] As will be readily understood, early clinical trials of new compounds generally only enroll subjects for which prior standard of care therapies, if available, have failed. Accordingly, in some instances, a subject treated in a trial setting can be naive to an engineered IL- 18 polypeptide (i.e., “engineered IL-18 naive”), but not necessarily naive to prior treatment for their condition, including e.g., cancer therapy. In later trials and following approval, drugs can, in some instances, supplant a prior standard of care and become a first-administered agent for one or more conditions, such that the drug, e.g., an engineered IL- 18, can be administered as a first treatment for a subject newly diagnosed with a condition, such as a cancer, solid tumor, or any non-limiting example listed herein.

[0082] In some instances, the subject receiving IV administration of an engineered IL- 18 has previously been treated with or otherwise previously received an engineered IL-18 polypeptide. In some instances, methods of the present disclosure can include testing for a likelihood of developing an ISR to the engineered IL-18 if administered subcutaneously. In some instances, a biological sample obtained from the subject can be used in such testing.

[0083] Other agents useful in the methods of the present disclosure, e.g., besides an engineered IL- 18 polypeptide, can be administered by various RO As. In some instances, the engineered IL- 18 and one or more other agents can be administered by the same ROA, i.e., intravenously. In some instances, the engineered IL- 18 and one or more other agents can be administered by different ROAs, i.e., IV engineered IL-18 administration and administration of the one or more other agents by a ROA other than IL- 18. In some instances, employing the same ROA for administration of an engineered IL-18 and one or more other agents can provide certainadvantages, including e.g., a convenience for the subject in e.g., receiving multiple agents by an IV ROA.

[0084] The amount of an engineered IL- 18 administered to a subject can vary, including e.g., where the amount of agent administered to a subject in a single dose can range from about 1 pg / kg (of body weight of the subject) or less to about 3000 pg / kg or more, including e.g., from about 1 pg / kg to about 3000 pg / kg, about 5 pg / kg to about 3000 pg / kg, about 10 pg / kg to about 3000 pg / kg, about 15 pg / kg to about 3000 pg / kg, from about 20 pg / kg to about 3000 pg / kg, from about 30 pg / kg to about 3000 pg / kg, about 1 pg / kg to about 2500 pg / kg, about 5 pg / kg to about 2500 pg / kg, about 10 pg / kg to about 2500 pg / kg, from about 15 pg / kg to about 2500 pg / kg, from about 20 pg / kg to about 2500 pg / kg, from about 30 pg / kg to about 2500 pg / kg, about 1 pg / kg to about 2000 pg / kg, about 5 pg / kg to about 2000 pg / kg, about 10 pg / kg to about 2000 pg / kg, about 15 pg / kg to about 2000 pg / kg, from about 20 pg / kg to about 2000 pg / kg, from about 30 pg / kg to about 2000 pg / kg, about 1 pg / kg to about 1500 pg / kg, about 5 pg / kg to about 1500 pg / kg, about 10 pg / kg to about 1500 pg / kg, about 15 pg / kg to about 1500 pg / kg, from about 20 pg / kg to about 1500 pg / kg, from about 30 pg / kg to about 1500 pg / kg, about 1 pg / kg to about 1000 pg / kg, about 5 pg / kg to about 1000 pg / kg, about 10 pg / kg to about 1000 pg / kg, about 15 pg / kg to about 1000 pg / kg, from about 20 pg / kg to about 1000 pg / kg, from about 30 pg / kg to about 1000 pg / kg, about 1 pg / kg to about 500 pg / kg, about 5 pg / kg to about 500 pg / kg, about 10 pg / kg to about 500 pg / kg, about 15 pg / kg to about 500 pg / kg, from about 20 pg / kg to about 500 pg / kg, from about 30 pg / kg to about 500 pg / kg, about 1 pg / kg to about 250 pg / kg, about 5 pg / kg to about 250 pg / kg, about 10 pg / kg to about 250 pg / kg, about 15 pg / kg to about 250 pg / kg, from about 20 pg / kg to about 250 pg / kg, from about 30 pg / kg to about 250 pg / kg, etc. In some instances, the engineered IL- 18 polypeptide is administered at a dose of at least 1 pg / kg, at least 5 pg / kg, at least 10 pg / kg, at least 15 pg / kg, at least 20 pg / kg, at least 30 pg / kg, at least 60 pg / kg, at least 120 pg / kg, at least 240 pg / kg, at least 360 pg / kg, at least 480 pg / kg, at least 600 pg / kg, at least 720 pg / kg, at least 840 pg / kg, at least 960 pg / kg, at least 1080 pg / kg, or at least 1200 pg / kg or more.

[0085] Where administered as multiple doses, the frequency of the administration of engineered IL-18 polypeptides can vary. For example, in some embodiments, each dose of successive doses of an engineered IL-18 composition is administered to the subject weekly, about every 7 days, at least 6 days after a previous dose of the composition, or at most 9 days after a previous dose. In some embodiments, each dose of successive doses of an engineered IL-18 composition is administered to the subject less frequently than weekly, including e.g., once every 2 weeks, once every three weeks, or once monthly.

[0086] In some embodiments, a dose of the engineered IL- 18 composition comprises at least about 1 pg / kg, at least about 5 pg / kg, at least about 10 pg / kg, at least about 15 pg / kg, at least about 20 pg / kg, at least about 30 pg / kg, at least about 90 pg / kg, at least about 180 pg / kg, at least about 360 pg / kg, at least about 600 pg / kg, at least about 900 pg / kg, or at least about 1200 pg / kg of the polypeptide per body weight of the subject. In some embodiments, each dose of the engineered IL-18 composition comprises between about 1 pg / kg to about 1200 pg / kg, between about 10 pg / kg to about 1200 pg / kg, between about 1 pg / kg to about 30 pg / kg, between about 10 pg / kg to about 30 pg / kg, between about 15 pg / kg to about 1200 pg / kg, between about 15 pg / kg to about 30 pg / kg, about 30 pg / kg to about 90 pg / kg, about 90 pg / kg to about 180 pg / kg, about 180 pg / kg to about 360 pg / kg, about 360 pg / kg to about 600 pg / kg, about 600 pg / kg to about 900 pg / kg, or about 900 pg / kg to about 1200 pg / kg of the polypeptide per body weight of the subject. In some embodiments, a dose of the engineered IL-18 composition comprises about 1 pg / kg, about 10 pg / kg, about 15 pg / kg, about 30 pg / kg, about 60 pg / kg, about 90 pg / kg, about 120 pg / kg, about 180 pg / kg, about 240 pg / kg, about 360 pg / kg, about 600 pg / kg, about 900 pg / kg, or about 1200 pg / kg of the polypeptide per body weight of the subject.

[0087] In some embodiments, each dose of the engineered IL- 18 composition comprises between about 60 pg / kg to about 1200 pg / kg, about 60 pg / kg to about 90 pg / kg, about 60 pg / kg to about 120 pg / kg, about 60 pg / kg to about 180 pg / kg, about 60 pg / kg to about 240 pg / kg, about 60 pg / kg to about 360 pg / kg, about 60 pg / kg to about 420 pg / kg, about 60 pg / kg to about 480 pg / kg, about 60 pg / kg to about 540 pg / kg, about 60 pg / kg to about 600 pg / kg, about 60 pg / kg to about 660 pg / kg, about 60 pg / kg to about 720 pg / kg, about 60 pg / kg to about 800 pg / kg, about 60 pg / kg to about 860 pg / kg, about 60 pg / kg to about 920 pg / kg, about 60 pg / kg to about 980 pg / kg, about 60 pg / kg to about 1040 pg / kg, or about 60 pg / kg to about 1200 pg / kg.

[0088] In some embodiments, each dose of the engineered IL- 18 composition comprises between about 120 pg / kg to about 1200 pg / kg, about 120 pg / kg to about 180 pg / kg, about 120 pg / kg to about 240 pg / kg, about 120 pg / kg to about 360 pg / kg, about 120 pg / kg to about 420 pg / kg, about 120 pg / kg to about 480 pg / kg, about 120 pg / kg to about 540 pg / kg, about 120 pg / kg to about 600 pg / kg, about 120 pg / kg to about 660 pg / kg, about 120 pg / kg to about 720 pg / kg, about 120 pg / kg to about 800 pg / kg, about 120 pg / kg to about 860 pg / kg, about 120 pg / kg to about 920 pg / kg, about 120 pg / kg to about 980 pg / kg, about 120 pg / kg to about 1040 pg / kg, or about 120 pg / kg to about 1200 pg / kg.

[0089] Amounts administered and doses described herein can, in some instances reflect the amount used in initial dosing of engineered IL- 18 naive subjects, repeat dosing of previously dosed subject, or both. Accordingly, in some instances, the methods involve administering to subjects that have not been previously administered the engineered IL-18 polypeptide. In someembodiments, the methods involve administering an engineered IL- 18 polypeptide to subjects that have been previously administered the engineered IL- 18 polypeptide. In some embodiments, a dose described herein can be employed in initial dosing or repeat dosing. In some embodiments, repeat dosing can employ the same dose as used for initial dosing. In some embodiments, repeat dosing can employ one of more different doses as compared to what was employed for initial dosing.

[0090] In some instances, the dose administered to a subject can be escalated, e.g., during a course of therapy. In some instances, a subject can be administered a dose based on prior dose escalation studies (where successive doses administered to the subject, if administered, can vary). Subjects can exhibit a tolerance for a dose of the engineered IL- 18 composition comprising the polypeptide and can be administered a higher dose of the polypeptide in a subsequent dose of the successive doses of the engineered IL-18 composition, provided, e.g., that the tolerability of the escalated dose has already been demonstrated in other subjects. In some instances, doses are not escalated and all successive doses administered, or some potion thereof, are the same. For example, in some instances, a subject is administered a set dose (e.g., about 1 pg / kg, about 10 pg / kg, about 15 pg / kg, about 30 pg / kg, about 60 pg / kg, about 90 pg / kg, about 120 pg / kg, about 180 pg / kg, about 240 pg / kg, about 360 pg / kg, etc.) at a regular frequency (e.g., weekly, every two weeks, etc.) for some period of time (e.g., 4 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, 28 weeks, 32 weeks, 36 weeks, 40 weeks, 44 weeks, 48 weeks, 52 weeks, etc.)

[0091] In some embodiments, a subsequent dose of the engineered IL- 18 composition comprises a greater amount of the polypeptide per body weight of the subject than an amount of the polypeptide administered previously to the subject. In some embodiments, the previous amount of the polypeptide was tolerated by the subject.

[0092] In some embodiments, a subsequent dose of the engineered IL- 18 composition comprises a greater amount of the polypeptide per body weight of the subject than an amount of the polypeptide in a previous dose of the engineered IL- 18 composition. In some embodiments, the previous dose was tolerated by the subject.

[0093] In some embodiments, a subsequent dose of the engineered IL- 18 composition comprises a same or similar amount of the polypeptide per body weight of the subject to an amount of the polypeptide administered previously to the subject. In some embodiments, the amount of the polypeptide is associated with a recorded Treatment-Emergent Adverse Event (TEAE) in the subject. In some embodiments, the recorded TEAE is a Grade 1, Grade 2, Grade 3, or Grade 4 TEAE according to U.S. National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0, U.S. Department of Health and Human Services,published November 27, 2027. In some embodiments, the recorded TEAE is associated with no limitation to mild limitation in activity of the subject. In some embodiments, a subsequent dose of the engineered IL- 18 composition comprises a same or similar amount of the polypeptide per body weight of the subject to an amount of the polypeptide in a previous dose of the engineered IL- 18 composition. In some embodiments, the previous dose is associated with causing a recorded TEAE in the subject. In some embodiments, the recorded TEAE is a Grade 1, Grade 2, Grade 3, or Grade 4 TEAE according to NCI CTCAE Version 5.0. In some embodiments, the recorded TEAE is associated with no limitation to mild limitation in activity of the subject. In some embodiments, the recorded TEAE is not associated with the need to provide medical intervention, assistance, or therapy to the subject. In some embodiments, the previous dose is a highest tolerable dose. In some embodiments, a subsequent dose of the engineered IL- 18 composition comprises a lesser amount of the polypeptide per body weight of the subject than an amount of the polypeptide administered previously to the subject. In some embodiments, the subject did not tolerate the amount of the polypeptide administered previously. In some embodiments, the amount of the polypeptide administered previously is associated with dose limiting toxicity (DLT). In some embodiments, the subsequent dose is a previously tolerated dose.

[0094] In some embodiments, a subsequent dose of the engineered IL- 18 composition comprises a lesser amount of the polypeptide per body weight of the subject than an amount of the polypeptide in a previous dose of the engineered IL- 18 composition. In some embodiments, the previous dose was not tolerated by the subject. In some embodiments, the previous dose is associated with dose limiting toxicity (DLT). In some embodiments, the subsequent dose is a previously tolerated dose.

[0095] In some embodiments, engineered IL- 18 composition (as a monotherapy or in combination with other therapeutics) are administered for at least 24 weeks. In some embodiments, the engineered IL- 18 compositions (as a monotherapy or in combination with other therapeutics) are administered until the subject has been administered at least 8 doses. In some embodiments, the engineered IL- 18 compositions (as a monotherapy or in combination with other therapeutics) are administered until the subject exhibits a complete response (CR), has been treated for at least 24 weeks, has been administered at least 8 doses, or some combination thereof. In some instances, engineered IL- 18 compositions are administered (or administration is continued) regardless of subjects’ response or achievement of stable disease, including e.g., where such continued treatment is continued for months or years, including e.g., until the subject progresses.

[0096] In some embodiments, engineered IL- 18 compositions (as a monotherapy or in combination with other therapeutics) are administered until the subject exhibits an intervention- related toxicity specified as a reason for permanent discontinuation. In some embodiments, the intervention-related toxicity is at least one of a Grade 4, Grade 3, or recurrent Grade 2 TEAE according to NCI CTCAE Version 5.0. In some embodiments, the TEAE is associated with marked or severe limitation to extreme limitation in activity of the subject. In some embodiments, the TEAE is life threatening or requires significant medical intervention or hospitalization. In some embodiments, the engineered IL- 18 compositions are administered (as a monotherapy or in combination with other therapeutics) for at most about 2 years. In some instances, engineered IL- 18 compositions are administered (or administration is continued) for more than 2 years, including e.g., where administration is continued regardless of a subject’s response or achievement of stable disease, including e.g., where such continued treatment is continued for months or years, including e.g., for more than 2 years or until the subject progresses.

[0097] Compositions that include an engineered IL-18 polypeptide, solely or present with one or more additional active agents, useful in the herein described methods are administered intravenously and can thus be correspondingly prepared, packaged, or sold in formulations suitable for intravenous administration. For example, in some instances, an engineered IL- 18 can be formulated together with an additional cancer therapeutic, such as e.g., ICI, including where the two components are formulated together in a single container suitable for IV delivery in unit dosage form. In some instances, an engineered IL- 18 can be provided together with an additional cancer therapeutic as a part of a treatment kit, including e.g., where the two components are present in the kit in separate containers configured to be co-administered as part of a combination therapy, where the engineered IL- 18 is configured to be administered by IV and the additional cancer therapeutic may be configured to be administered by IV (together with or separate from the engineered IL- 18 IV administration) or by a ROA other than IV. In some instances, an engineered IL- 18 can be provided together with a B cell depleter as a part of a treatment kit, including e.g., where the two components are present in the kit in separate containers configured to be administered as part of a treatment protocol requiring pre-treatment with the B cell depleter, where the engineered IL- 18 is configured to be administered by IV and the B cell depleter may be configured to be administered by any appropriate ROA, such as e.g., by IV (separate from the engineered IL- 18 IV administration).

[0098] Compositions other than, and administered separately from, engineered IL- 18 compositions, including e.g., other cancer agent compositions (e.g., ICIs, chemotherapeutics, etc.), agents for CRS prophylaxis, and the like, useful in the herein described methods can beadministered by (and be correspondingly prepared, packaged, or sold in formulations suitable for) essentially any suitable route of administration (ROA), including parenteral or enteral routes including e.g., oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, intravenous, transdermal, intralesional, subcutaneous, intramuscular, ophthalmic, intrathecal and other known RO As.

[0099] Other contemplated formulations include projected nanoparticles, liposomal preparations, lipid nanoparticles, and other preparations containing the active ingredient. In some instances, the methods of the present disclosure include administering the engineered IL- 18 polypeptide by an intravenous ROA and one or more other agents via parenteral administration. In some instances, the methods of the present disclosure include administering via intravenous administration the engineered IL- 18 polypeptide and any other administered agent, including all other administered agents.

[0100] In some instances, subjects in the herein described methods will not receive or will be excluded from other treatments, e.g., treatments besides those involving administration of an engineered IL- 18 polypeptide. For example, in some instances, a subject treated according to methods described herein can abstain from certain other treatments, including e.g., abstaining from antihypertensive drugs, including e.g., where such a subject abstains for at least 24 hours before and at least 48 hours after administration of the engineered IL-18. Accordingly, subjects of the herein described methods can, but need not necessarily, include subjects that have not been administered antihypertensive drugs, including e.g., having not been administered antihypertensive drugs for at least 24 hours before administration of an engineered IL-18 polypeptide. In some instances, abstaining from antihypertensive drugs relates only to the days of treatment and subjects otherwise maintain standard medication regimens, e.g., outside of about 24 hours before and about 48 hours after administration of the engineered IL- 18 polypeptide.

[0101] In some instances, methods of the present disclosure can further include administering one or more other agents (e.g., agents besides an engineered IL-18 polypeptide), including e.g., where such other agents can include an intravenous glucocorticoid, acetaminophen, antihistamine, or a combination thereof.

[0102] In some instances, methods of the present disclosure can include the administration of one or more agents to prevent cytokine release syndrome (CRS) in the subject, referred to herein as CRS prophylaxis. CRS prophylaxis can prevent or lessen the risk of CRS that may otherwise occur following the administration of an engineered IL-18 polypeptide, such as a DR IL-18 polypeptide. CRS prophylaxis can reduce the display of symptoms of CRS, including e.g., where such a reduction is as compared to a comparable subject that was administered at least acomparable amount of engineered IL-18 polypeptide but did not receive CRS prophylaxis. An agent administered for CRS prophylaxis may be referred to as a CRS prophylactic agent.

[0103] Non-limiting examples of symptoms of CRS include fever, chills, tiredness, nausea, vomiting, diarrhea, headache, cough, low blood pressurejoint pain, muscle pain, skin rash, shortness of breath, confusion, dizziness, difficulty swallowing, increased heart rate, reduced heart function, irregular heartbeat, organ failure, and swelling.

[0104] In some embodiments, administration of CRS prophylaxis will prevent the onset or progression of CRS in a subject administered composition comprising an engineered IL-18 polypeptide, including e.g., at least about 30 pg / kg, at least about 60 pg / kg, or more of an engineered IL- 18 polypeptide. For example, administration of CRS prophylaxis will prevent the onset of or progression to grade 5 CRS, grade 4 CRS or higher, and / or grade 3 CRS or higher, including e.g., where CRS grading is performed according to Consensus American Society for Transplantation and Cellular Therapy (ASTCT) grading for CRS as described in Lee et al. Biology of Blood and Marrow Transplantation. 25 (2019) 625 - 639. In some instances, a lack of onset or progression of CRS may be indicated by a lack of fever (temp, greater than or equal to 38 deg. C), a lack of hypotension, hypotension that does not require vasopressors, hypotension requiring only one vasopressor with or without vasopressin, a lack of hypoxia, hypoxia requiring only low-flow nasal canula or blow-by, hypoxia requiring high-flow nasal- canula (or facemask, nonrebreather mask, or venturi mask) but not requiring positive pressure, or a combination thereof.

[0105] In some embodiments, CRS prophylaxis in a subject administered a therapy that comprises administration of a composition comprising an engineered IL- 18 polypeptide, including e.g., at least about 30 pg / kg, at least about 60 pg / kg, or more of an engineered IL-18 polypeptide, will negate the need for later treating the subject for CRS. For example, in some instances, CRS prophylaxis in such scenarios will mean that administration of immunosuppressants, such as, e.g., corticosteroids (e.g., glucocorticoids), anti-interleukin 6 (IL- 6) agents (e.g., tocilizumab, siltuximab, clazakizumab), TNF-a signaling inhibitors (e.g., etanercept), T cell-depleting antibodies (e.g., alemtuzumab), IL-lR-based inhibitors (e.g., anakinra), cyclophosphamide, Bruton’s tyrosine kinase (BTK) inhibitors (e.g., ibrutinib), combinations thereof, and the like will not be necessary and will not be administered. In some embodiments, a subject receiving CRS prophylaxis and administered a therapy that comprises administration of an engineered IL-18 polypeptide, including e.g., at least about 30 pg / kg, at least about 60 pg / kg, or more of a DR-IL-18 polypeptide, will not require and will not be administered an immunosuppressant to treat CRS. For example, in some instances, such a subject will not be administered any corticosteroids (e.g., glucocorticoids) and / or any anti-interleukin 6 (IL-6) agents during a cycle of the combination therapy or during the entire course of the combination therapy.

[0106] A CRS prophylactic agent can be administered to a subject at any convenient time before the onset of CRS, and, in some instances, also concurrently with and / or continuing into a CRS episode. A CRS prophylactic agent can be administered concurrently with, before, and / or following administration of one or more doses of an engineered IL- 18 polypeptide. In some instances, a CRS prophylactic agent is administered before or with a subject’s first dose of the engineered IL-18 polypeptide. In some embodiments, the CRS prophylactic agent is administered as premedication and / or post-dose medication to a dose of the engineered IL-18 polypeptide. In some embodiments, the method further comprises administering a dose of a CRS prophylactic agent to the subject with or before a dose of one or more successive (i.e., repeat) doses of the engineered IL-18 polypeptide. In some embodiments, the method comprises administering a dose of a CRS prophylactic agent to the subject with or before each dose of the engineered IL-18 polypeptide, including e.g., each successive dose, or the first dose and each successive dose of the engineered IL-18 polypeptide.

[0107] Agents useful as CRS prophylactic agents include steroids, non-steroidal antiinflammatory drugs (NSAID), acetaminophen, diphenhydramine, histamine Hl antagonists, famotidine, H2 blockers, supplemental fluids, and combinations thereof. Useful CRS prophylactic agents are described in more detail below.

[0108] Any useful route of administration (ROA) can be employed for delivering a CRS prophylactic agent or combination of agents, including e.g., where the agent(s) are administered by a parenteral or enteral route, including e.g., orally, subcutaneously, intramuscularly, intravenously, intrathecally, rectally, vaginally, or nasally. In some instances, a combination of CRS prophylactic agents can be employed and the combination can be administered through the same ROA or through multiple different RO As. The timing of administration of a CRS prophylactic agent, or combination of agents, can vary, including where the agent(s) are administered, e.g., hourly, about every several hours, twice daily, daily, twice a week, every six days, weekly, every two weeks, every three weeks, monthly, every two months, every three months, every four months, every six months, once a year. In some instances, the administration of a CRS prophylactic agent, or combination of agents, can be continued for the entire course of therapy with an engineered IL- 18 polypeptide, including e.g., where the administration occurs at some frequency (including those frequencies listed above) for the entire course of treatment. CRS prophylactic agent administration need not necessarily occur at a regular frequency and can, in some instances, occur at irregular, intermittent or discontinuous intervals. In some instances, the CRS prophylactic agent, or combination of agents, can be administered only inconjunction with (i.e., before, concurrently, or following) the first dose of engineered IL-18 polypeptide. In some instances, the CRS prophylactic agent, or combination of agents, can be administered in conjunction with (i.e., before, concurrently, or following) each dose of engineered IL- 18 polypeptide.

[0109] Administration of one or more CRS prophylactic agents can vary and can include e.g., where such agents are administered not more than 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, or 72 hours prior to receiving a dose of an engineered IL- 18. In some instances, one or more CRS prophylactic agents can be administered at the same time, immediately before, or immediately after administration of a dose of an engineered IL-18. In some instances, one or more CRS prophylactic agents can continue to be administered following the dose of an engineered IL-18, including e.g., for at least 6 hours, at least 12 hours, at least 24 hours, at least 36 hours, at least 48 hours, or at least 72 hours following the dose of an engineered IL-18.

[0110] Any of the herein described groups of CRS prophylactic agents and individual CRS prophylactic agents and any of the herein described method of using such groups and individual CRS prophylactic agents may be combined into a useful CRS prophylaxis combination.[OHl] For example, in some embodiments, CRS prophylaxis administered to a subject will include at least a NSAID and at least a histamine Hl antagonist; at least acetaminophen and at least a histamine Hl antagonist; at least a NSAID and at least acetaminophen; or at least a NSAID, at least a histamine Hl antagonist, and at least acetaminophen. Such NSAID, histamine Hl antagonist, and acetaminophen used in such combinations may be administered according to the administration and / or dosing of each agent as described herein. In some embodiments, CRS prophylaxis administered to a subject will include at least indomethacin or ibuprofen and at least diphenhydramine; at least acetaminophen and at least diphenhydramine; at least indomethacin or ibuprofen and at least acetaminophen; or at least indomethacin or ibuprofen, at least diphenhydramine, and at least acetaminophen. Such indomethacin or ibuprofen, diphenhydramine, and acetaminophen used in such combinations may be administered according to the administration and / or dosing of each agent as described herein. In some embodiments, such combinations will further include an H2 blocker, including e.g., where the H2 blocker used is famotidine and the famotidine is administered according to the administration and / or dosing described herein.

[0112] In some embodiments, CRS prophylaxis will include at least an NSAID, a histamine Hl antagonist, and acetaminophen, including e.g., where (i) the NSAID is oral indomethacin administered at 50 mg to 100 mg per day (inc., e.g., at 75 mg per day), or oral ibuprofen administered at 600 mg to 2,400 mg per day (inc. e.g., at 800 mg to 1,800 mg per day); (ii) the histamine Hl antagonist is intravenous or oral diphenhydramine administered at a dose of 25 mgto 100 mg (inc. e.g., a dose of 50 mg); and (iii) the acetaminophen is oral acetaminophen administered at 350 mg to 4,000 mg per day (inc. e.g., at 650 mg to 3,900 mg per day). In some embodiments, such a combination will further include an H2 blocker, including e.g., where the H2 blocker used is famotidine and the famotidine is administered according to the administration and / or dosing described herein.

[0113] In some embodiments, a CRS prophylaxis combination described herein will reduce the display of one or more symptoms of CRS, including reducing the one or more symptoms as compared to a subject administered the same treatment protocol without the CRS prophylaxis. In some embodiments, a CRS prophylaxis combination described herein, such as the combination described above, will prevent a subject with cancer, treated with a therapy that includes IV administration of a composition comprising an engineered IL- 18 polypeptide comprising an amino acid sequence set forth in any one of SEQ ID NOs: 7, 17-22, 55-57, and 62 (such as, e.g., SEQ ID NO: 20), from experiencing one or more (including all) symptoms of grade 3 or higher CRS or grade 4 CRS. In some embodiments, a CRS prophylaxis combination described herein, such as the combination described above, will prevent a subject with cancer, treated with a therapy that includes IV administration a composition comprising a polypeptide that is an engineered IL- 18 polypeptide comprising an amino acid sequence set forth in any one of SEQ ID NOs: 7, 17-22, 55-57 (such as, e.g., SEQ ID NO:20), from requiring treatment for CRS, including treatment with one or more (or any) immunosuppressants, such as e.g., a corticosteroid, an anti-interleukin 6 (IL-6) agents (such as e.g., tocilizumab), or any combination thereof.

[0114] In some embodiments, methods of the present disclosure include pretreatment of subjects (i.e., prior to the administration of engineered IL-18 polypeptides to the subject) with a B-cell depleting agent, referred to herein as a “B-cell depleter”, to deplete the B-cells in the subject. A B-cell depleter provides a means for depleting the B-cells of a subject. Without being bound by theory, pretreatment to deplete the B cells of a subject may reduce or suppress certain immune responses of the subject directed to delivered reagents, such as e.g., engineered IL- 18 polypeptides, and subsequently reduce or suppress the development of anti-drug antibodies (ADAs), neutralizing antibodies (nAbs), and the like, inc. e.g., engineered IL- 18 polypeptide AD As, engineered IL-18 polypeptide nAbs, and the like. In some instances, both the B-cell depleter and the engineered IL-18 polypeptide are administered by an IV ROA.

[0115] The term “B-cell depleter” refers to a B-cell depleting agent that is useful in reducing or eliminating B cells and / or B cell production in a subject. Useful B-cell depleting agents can function independently or in combination with other agents to deplete B cells and include agentsof various functional modalities, such as e.g., small and large molecules, including e.g., antibody B-cell depleting agents. Non-limiting examples of B-cell depleters are provided below.

[0116] In some instances, methods of the present disclosure include pretreating a subject by administering to the subject, prior to or concurrent with administration of an engineered IL- 18 polypeptide, a B-cell depleter, such as e.g., a B-cell depleting antibody. B cell depleters, including B-cell depleting antibodies, can directly target B-cells and / or B-cell progenitors of a subject, including e.g., by directly targeting an epitope of a protein expressed by B cells and / or B-cell progenitors, such as, e.g., CD20. For example, in some instances, methods of the present disclosure include pretreating a subject by administering to the subject a B-cell depleting antibody that is an anti-CD20 antibody, such as e.g., obinutuzumab.

[0117] Pre-treatment for B-cell depletion can include administering one or more doses of a B- cell depleter, including e.g., where the pre-treating comprises administering the B-cell depleter in a single dose, or where the pre-treating comprises administering the B-cell depleter in multiple doses, such as e.g., two separate doses. Where multiple separate doses are employed, the doses can be administered essentially concurrently or separated by a time period, where such a time period can vary and can range from at least 1 hour to over 24 hours. In some instances, separate doses of a B-cell depleter are separated by at least 2 hours, at least 4 hours, at least 6 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 14 hours, at least 16 hours, at least 18 hours, at least 20 hours, at least 22 hours, or at least 24 hours. In some instances, separate doses of a B-cell depleter, while separate by some time period, are nonetheless administered within 72 hours, including e.g., within 48 hours, within 36 hours, or within 24 hours. In some instances, separate doses are administered on consecutive days, including e.g., two doses administered on consecutive days, including e.g., at least 6 or 12 hours apart on consecutive days.

[0118] The administered dose(s) of B-cell depleter to achieve B-cell depletion as employed in the herein described methods can vary, e.g., depending on the particular B-cell depleter employed and, e.g., whether the agent is administered in one dose or multiple doses. For example, in some instances, a subject is pre-treated by being administered at least 2000 mg of obinutuzumab, including e.g., where the obinutuzumab is administered in one or two doses.

[0119] The timing of pretreatment, relative to the administration of a dose of an engineered IL-18 polypeptide, can vary. In some instances, the pretreatment is performed at least 4 days prior to administering the engineered IL-18 polypeptide, including e.g., at least 5 days prior, at least 6 days prior, at least 7 days prior, at least 8 days prior, at least 9 days prior, at least 10 days prior, at least 11 days prior, at least 12 days prior, at least 13 days prior, or at least 14 days prior. Where a single dose of B-cell depleter is employed, the pretreatment is “performed” uponadministration of the dose. Where multiple doses of B-cell depleter are employed, the pretreatment is “performed” upon administration of the final. In some instances, the pretreatment is performed no more than 15 days prior to administering the engineered IL-18 polypeptide, including e.g., no more than 14 days prior to administering the engineered IL-18 polypeptide, no more than 13 days prior to administering the engineered IL- 18 polypeptide, no more than 12 days prior to administering the engineered IL-18 polypeptide, no more than 11 days prior to administering the engineered IL- 18 polypeptide, no more than 10 days prior to administering the engineered IL-18 polypeptide, no more than 9 days prior to administering the engineered IL-18 polypeptide, no more than 8 days prior to administering the engineered IL-18 polypeptide.

[0120] In some instances, pre-treatment to deplete the B-cells of a subject prior to administering an engineered IL- 18 can include intravenous administration of the B-cell depleter. In some instances, the B-cell depleter and the engineered IL-18 can be administered by different RO As. In some instances, the B-cell depleter and the engineered IL-18 can be administered by the same ROA. In some instances, employing the same ROA for administration of a B-cell depleter and an engineered IL-18 can provide certain advantages, including e.g., a convenience for the subject in e.g., receiving both agents by an IV ROA.

[0121] In some instances, depletion of B-cells in a subject following pretreatment can be confirmed, e.g., prior to administering a dose (including an initial dose or a next dose) of an engineered IL-18. In some instances, levels of B-cell depletion can be monitored, e.g., before pretreatment, during pretreatment, following pretreatment, before engineered IL- 18 administration, during IL-18 administration, following IL-18 administration, during the entire course of treatment (including or excluding pretreatment), after the course of treatment, during one or more particular timepoints of the course of treatment, or any combination thereof. Various methods for measuring or monitoring B-cell levels and / or B-cell depletion can be employed, including those that measure B-cell levels directly or indirectly, such as e.g., cell counts (including e.g., white blood cell counts), flow cytometry, microscopy, mass cytometry, immunoassays (e.g., ELISA), and the like.

[0122] Methods of the present disclosure may be performed with or without obinutuzumab pretreatment. In some instances, obinutuzumab pre-treatment consists of 2 doses given one day apart, and via IV dosing. In some instances, adult test subjects with solid tumors who have developed disease progression through standard therapy, or for whom standard of care therapy that prolongs survival is unavailable or unsuitable, are given obinutuzumab pre-treatment and pre-medicated for CRS prior to dosing with an engineered IL-18 polypeptide. In some instances, adult test subjects with solid tumors who have developed disease progression through standardtherapy, or for whom standard of care therapy that prolongs survival is unavailable or unsuitable, are not administered obinutuzumab pre-treatment.

[0123] In some embodiments, subjects pretreated with obinutuzumab will receive 2 x 1000 mg of obinutuzumab, administered on 2 consecutive days (e.g., Day -7 and Day -6) before Cycle 1, Day 1 (C1D1) engineered IL-18 polypeptide (note: the treatment naming convention of “C#D#” represents cycle and day numbers respectively). Dosing with the engineered IL- 18 polypeptide will be performed at least 7 days after the first dose of obinutuzumab and, in some cases, up to 14 days after the first dose of obinutuzumab.

[0124] In some embodiments, pre-engineered IL-18 polypeptide and post-obinutuzumab medication, including for CRS prophylaxis, may include: administering an oral nonsteroidal anti-inflammatory drug (NS AID) 1-2 hours prior to therapy and continuing for at least 48 hours post-dose. Choice of NS AID can include but not be limited to indomethacin 25 mg 3 times daily or oral ibuprofen 200 to 600 mg every 6 to 8 hours; adding oral acetaminophen 650 mg every 4 to 6 hours as needed for the first 24 hours post-dose; for test subjects with contraindication to NSAIDs, oral acetaminophen is administered 650 mg every 4 to 6 hours beginning 1 hour prior to engineered IL- 18 polypeptide dosing and continued for at least 48 hours post-dose; administering IV or oral diphenhydramine 50 mg or a comparable dose of a next-generation histamine Hl antagonist 30-60 minutes prior to dosing with the engineered IL-18 polypeptide; for test subjects with or at risk for hypotension, administering of up to 3 liters of oral / IV fluids in the 24 hours post-dose; where indicated for subsequent doses, administering famotidine (or other H2 blocker) at 20 mg IV or 20-40 mg orally, 30-60 minutes prior to dosing with the engineered IL- 18 polypeptide.

[0125] In some embodiments, a 1stdose obinutuzumab regimen to reduce obinutuzumab infusion-related reactions (IRRs), will include administering IV glucocorticoid (e.g., 20 mg dexamethasone or 80 mg methylprednisolone) >1 hour prior to obinutuzumab infusion and administering 650-1000 mg acetaminophen and antihistamine (e.g., 50 mg diphenhydramine) >30 minutes before obinutuzumab infusion.

[0126] In some embodiments, a 2nddose obinutuzumab regimens to reduce obinutuzumab IRRs will include: administering 650-1000 mg acetaminophen >30 minutes before obinutuzumab infusion; or administering 650-1000 mg acetaminophen and antihistamine (e.g., 50 mg diphenhydramine) >30 minutes before obinutuzumab infusion for subjects experiencing an IRR (Grade 1-2) with the previous infusion; or administering IV glucocorticoid (e.g., 20 mg dexamethasone or 80 mg methylprednisolone), 650-1000 mg acetaminophen, and antihistamine (e.g., 50 mg diphenhydramine) >1 hour prior to obinutuzumab infusion for subjects with a Grade3 IRR with the previous infusion or with a lymphocyte count >25 x 109 / L prior to next treatment.

[0127] In some embodiments, initial pre-treatment with obinutuzumab will include: 1000 mg daily IV x 2 (on 2 consecutive days), with the first dose given at least 7 days (and up to 14 days) prior to first dose of the engineered IL- 18 polypeptide. Depending on the initial tolerability of this dosing regimen, a single dose of obinutuzumab of 2000 mg can be used. The dosing interval between the 2 doses of obinutuzumab, and between the second dose of obinutuzumab and the first dose of the engineered IL-18 polypeptide can be lengthened depending on the initially observed tolerability profile. If B-cell depletion is determined to be clinically inadequate, additional doses of obinutuzumab can be used.

[0128] In some embodiments, a first dose (Day -7 prior to C1D1) obinutuzumab regimen will include a 1000 mg dose administered at 50 mg / hr, with optional escalation in 50 mg / hr increments every 30 minutes to a maximum of 400 mg / hr.

[0129] In some embodiments, a second dose (Day -6 prior to C1D1) obinutuzumab regimen will include a 1000 mg dose. If no IRR or an IRR of Grade 1 occurred during the previous infusion and the final infusion rate was 100 mg / hr or faster, infusions are started at a rate of 100 mg / hr and increased by 100 mg / hr increments every 30 minutes to a maximum of 400 mg / hr. If an IRR of Grade 2 or higher occurred during the previous infusion, infusions are administered at 50 mg / hr. Optionally, the rate of infusion is escalated in increments of 50 mg / hr every 30 minutes to a maximum rate of 400 mg / hr.

[0130] In some instances, methods of the present disclosure can further include administering one or more additional anti -cancer agents (i.e., in addition to the administered engineered IL- 18). Essentially any anti-cancer agent can be employed, including e.g., chemotherapeutics, immune checkpoint inhibitors (ICIs), opsonizing agents, tumor-targeting antibodies, CAR T- cells, CAR NK cells, oncolytic viruses, and the like. Selection of additional anti-cancer agents to be used in combination with an engineered IL- 18 polypeptide can depend on factors such as the type of cancer to be treated, the stage of cancer progression, prior treatments, etc.

[0131] Methods of the present disclosure find use in subject for the treatment of disease, such as but not limited to e.g., cancer (including e.g., solid tumor cancers), infections (including e.g., viral infections, bacterial infections, parasitic infections, etc.), diseases related to decreased or repressed immunity (including e.g., neutropenia, e.g., as a result of chemotherapy), genetic and metabolic diseases, and the like.

[0132] In some instances, subjects treated in the herein described methods have not been previously administered the engineered IL- 18 polypeptide. In some instances, subjects treated in the herein described methods have been previously administered the engineered IL- 18polypeptide (i.e., the same engineered IL-18). In In some instances, subjects treated in the herein described methods have not been previously administered a different engineered IL- 18 polypeptide and / or wt IL-18. In some instances, subjects treated in the herein described methods have not been previously treated with a checkpoint inhibitor, wherein such subjects may be referred to as checkpoint-naive subjects. In some instances, subjects treated in the herein described methods have received prior treatment for cancer and the methods of the present disclosure can find use in treating recurrent cancer (including e.g., local recurrent cancer, regional recurrent cancer, distant recurrent cancer), treatment resistant cancer, second cancers, and the like. In some instances, subjects treated in the herein described methods have a platinum-resistant cancer, such as e.g., platinum-resistant ovarian cancer.

[0133] The following are non-limiting examples of cancers that can be treated or prevented by the methods and compositions of the disclosure: acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, appendix cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and spinal cord tumors, brain stem glioma, brain tumor, breast cancer, bronchial tumors, Burkitt lymphoma, carcinoid tumor, central nervous system atypical teratoid / rhabdoid tumor, central nervous system embryonal tumors, central nervous system lymphoma, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, cerebral astrocytoma / malignant glioma, cervical cancer, childhood visual pathway tumor, chordoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous cancer, cutaneous t- cell lymphoma, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, Ewing family of tumors, extracranial cancer, extragonadal germ cell tumor, extrahepatic bile duct cancer, extrahepatic cancer, eye cancer, fungoides, gallbladder cancer, gastric (stomach) cancer, gastrointestinal cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (gist), germ cell tumor, gestational cancer, gestational trophoblastic tumor, glioblastoma, glioma, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, hypothalamic and visual pathway glioma, hypothalamic tumor, intraocular (eye) cancer, intraocular melanoma, islet cell tumors, Kaposi sarcoma, kidney (renal cell) cancer, Langerhans cell cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, lip and oral cavity cancer, liver cancer, lung cancer, lymphoma, macroglobulinemia, malignant fibrous histiocytoma of bone and osteosarcoma, medulloblastoma, medulloepithelioma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer with occult primary, mouth cancer, multiple endocrine neoplasia syndrome, multiple myeloma, mycosis, myelodysplastic syndromes, myelodysplastic / myeloproliferative diseases, myelogenous leukemia, myeloid leukemia,myeloma, myeloproliferative disorders, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oral cavity cancer, oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, papillomatosis, paraganglioma, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal parenchymal tumors of intermediate differentiation, pineoblastoma and supratentorial primitive neuroectodermal tumors, pituitary tumor, plasma cell neoplasm, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, primary central nervous system cancer, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter cancer, respiratory tract carcinoma involving the nut gene on chromosome 15, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, sezary syndrome, skin cancer (melanoma), skin cancer (nonmelanoma), skin carcinoma, small cell lung cancer, small intestine cancer, soft tissue cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck cancer, stomach (gastric) cancer, supratentorial primitive neuroectodermal tumors, supratentorial primitive neuroectodermal tumors and pineoblastoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell cancer, transitional cell cancer of the renal pelvis and ureter, trophoblastic tumor, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, visual pathway and hypothalamic glioma, vulvar cancer, waldenstrom macroglobulinemia, and Wilms Tumor.

[0134] In some non-limiting examples, the cancer is defined not (or at least not exclusively) by the histological tissue of origin, but by its molecular features. For example, tumors with high tumor mutational burden (TMB), tumors with microsatellite instability (MSI), tumors that lack or have diminished surface expression of MHC Class I (e.g., due to deletion of Beta-2 microglobulin or Tapasin), or tumors that express high levels of IL-18BP. In some cases, those molecular features are determined through next-generation sequencing (e.g., using TMB / MSI). In some cases, those molecular features are determined through immunohistochemistry, immunofluorescence, or flow cytometry. Thus, non-limiting examples of cancers that can be treated or prevented by the methods and compositions of the disclosure include solid tumor cancers, liquid cancers, blood cancers, teratomas, sarcomas, and carcinomas.

[0135] In some instances, the methods include treating a subject for a liquid tumor, e.g., a blood cancer. Accordingly, in some instances, the cancer for which the subject is treated is not a solid tumor. In some instances, the blood cancer is a B-cell neoplasm, not a lymphoma, not nonHodgkin's lymphoma, or the like.

[0136] In some instances, the methods include treating a subject for a solid tumor. Accordingly, in some instances, the cancer for which the subject is treated is not a liquid cancer, not blood cancer, not a B-cell neoplasm, not a lymphoma, not non-Hodgkin's lymphoma, or the like.

[0137] In some instances, the subject is treated, according to a herein described method, for a cancer selected from: melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-high tumors, high tumor mutation burden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer. In some instances, the subject is treated for Melanoma, Merkel cell, renal cell carcinoma (RCC), urothelial, non-small lung cancer (NSCLC; with no epidermal growth factor receptor [EGFR], tropomyosin receptor kinase [TRK] receptor, or anaplastic lymphoma kinase [ALK] positive mutations / fusions), triple-negative breast cancer (TNBC), squamous cell carcinoma of head and neck (SCCHN), microsatellite instability high (MSI-Hi) tumors, high tumor mutation burden (Hi-TMB) or mismatch repair deficient, gastric, cervical, endometrial, cutaneous squamous, small cell lung, esophageal, hepatocellular carcinoma (HCC), platinum resistant ovarian cancer, or microsatellite-stable colorectal cancer.

[0138] In some instances, the melanoma treated is histologically confirmed unresectable stage III / IV metastatic melanoma, treated with prior regimens which included a checkpoint inhibitor and BRAF / MEK inhibitor (if BRAF V600E / K mutation is present).

[0139] In some instances, the RCC treated is advanced (not amenable to curative surgery or radiation therapy) or metastatic (American Joint Committee on Cancer stage IV) RCC; histologically confirmed RCC with a clear-cell component; and / or treated with prior treatment regimens including a PD-1 or programmed death-ligand 1 (PD-L1) checkpoint inhibitor.

[0140] In some instances, the TNBC treated is includes than 1% of cells testing positive for estrogen and progesterone receptors determined by using standard immunohistochemistry (IHC) and adherent to the American Society of Clinical Oncology (ASCO) / College of American Pathologists (CAP) guidelines; is human epidermal growth factor 2 negative as determined by local pathologist, using IHC or in situ hybridization; and / or treated with prior regimens which included chemotherapy and a checkpoint inhibitor. TNBC is diagnosed in a tumor which does not express estrogen receptor or progesterone receptor, is not human epidermal growth factor receptor 2 (HER2) 3+ on IHC or is negative by fluorescence in situ hybridization (FISH).

[0141] In some instances, the NSCLC treated includes a histologic diagnosis of metastatic NSCLC which is non-squamous or squamous with no EGFR, TRK receptor, ALK, ROS1 rearrangements, or BRAF positive mutations / fusions; and / or treated with prior regimens whichhave included a platinum-based chemotherapy and a checkpoint inhibitor (unless the subjects is ineligible for a checkpoint inhibitor).

[0142] In some instances, the SCCHN treated is histologically confirmed relapsed, metastatic SCCHN (oral cavity, oropharynx, larynx, or hypopharynx); recurrent / metastatic SCCHN not amenable to surgery or radiation therapy; previously treated with platinum-based chemotherapy and checkpoint inhibitor therapy; and / or previously treated with prior regimens which included chemotherapy or radiotherapy or checkpoint inhibitor therapy.

[0143] In some instances, the MSI-Hi Tumors treated include demonstration of microsatellite instability by local testing (polymerase chain reaction [PCR] or IHC); and / or treatment with prior regimens for locally advanced / metastatic disease. A MSI high tumor may have mutations in 30% or more microsatellites by PCR or be negative for MSH1 / 2 / 6 or PMS-2 by IHC.

[0144] In some instances, the Hi-TMB high tumor treated will have 10 mut / Mb or greater calculated from whole genome sequencing or whole exome sequencing.

[0145] Dosing of subject with an engineered IL-18 polypeptide will vary, as described herein, and can, in some instances, be determined based on monitoring of subjects. In some instances, dosing can be determined based on a modified toxicity probability interval (mTPI) design, including e.g., where dosing is performed according to precalculated decisions based on the mTPI. In some instances, doses can be escalated, within a treatment regimen of a single subject or across subjects or cohorts. In some instances, an initial dose of 1 pg / kg, 5 pg / kg, 10 pg / kg, 15 pg / kg, 20 pg / kg, 30 pg / kg, 60 pg / kg, 90 pg / kg, 120 pg / kg, 180 pg / kg, 240 pg / kg, 360 pg / kg, 600 pg / kg, 900 pg / kg, or 1200 pg / kg can be employed, e.g., as initial doses or a dose escalation regimen. In some instances, doses can be increased above 1200 pg / kg, including e.g., starting from 1200 pg / kg and increasing in increments of 33%, e.g., until DLT is observed.

[0146] IV administration of a dose of an engineered IL-18 polypeptide may be performed according to any convenient means. For example, in some instances, a dose of the engineered IL- 18 may be diluted into a standard IV bag, such as e.g., a 50 mL, 100 mL, 200 mL, 250 mL, 500 mL, of 1000 mL IV bag of an appropriate (e.g., pharmaceutically acceptable) diluent, such as saline and the mixture may be infused. In some instances, the IV bag may contain a volume of diluent substantially less than the maximum volume of the IV bag, including e.g., where the IV bag contains 100 mL or less, 50 mL or less, 25 mL or less, 20 mL or less, 15 mL or less, 10 mL or less, etc. In some embodiments, a composition comprising the engineered IL-18 polypeptide (e.g., at a concentration of 30 mg / mL) is diluted into a volume of diluent (e.g., 20 mL or less of saline) in an IV bag of a larger maximum volume (e.g., 50 mL or more) to prepare a patient dose of 10 pg / kg, 20 pg / kg, 30 pg / kg, 60 pg / kg, 120 pg / kg, or 240 pg / kg engineered IL-18 polypeptide and the prepared solution is infused. Suitable prepared (i.e., diluted, infusion-ready) preparation concentrations of engineered IL- 18 polypeptide will vary and may include e.g., where the final concentration is about 0.01 mg / mL or less to about 20 mg / mL or more, including e.g., 0.01 mg / mL to 20 mg / mL, 0.1 mg / mL to 20 mg / mL, 0.01 mg / mL to 10 mg / mL, 0.1 mg / mL to 10 mg / mL, 0.01 mg / mL to 5 mg / mL, 0.1 mg / mL to 5 mg / mL, 0.01 mg / mL to 2 mg / mL, , 0.1 mg / mL to 2 mg / mL, and the like. In some instances, the volume infused into a subject is dependent on the subject’s size. In some instances, an IV bag containing a volume of diluent (before addition of an engineered IL-18 polypeptide) is drained and refilled with a suitable volume for infusion and then the dose is prepared. In some instances, an IV bag containing more diluent than desired is partially drained until a desired volume of diluent is obtained, and then the dose is prepared.

[0147] In some instances, an IV pump (also sometimes referred to as an IV driver) may be employed to deliver the prepared IV solution (i.e., desired amount of diluent containing the desired dose of engineered IL- 18 polypeptide) to a subject at a desired rate. Any suitable IV pump may be employed for rate controlled IV infusion. For example, in some instances, the dose of the engineered IL- 18 polypeptide is infused over 2 hours or less using an infusion pump, and e.g., compatible IV bag, tubing, and other devices. In some instances, the dose of the engineered IL-18 polypeptide is infused over 2 hours or less using a syringe pump, and e.g., compatible syringe, tubing, and other devices. In some instances, if an initial infusion time (e.g., 2 hrs) is well tolerated by a subject, subsequent infusion times may be decreased (e.g., 1 hr or less).

[0148] Infusion during engineered IL- 18 administration may be facilitated by any convenient and appropriate means, such as e.g., gravity infusion, pump infusion (such as e.g., IV infusion pump, syringe pump infusion), or the like. One or more than one (e.g., 2, 3, 4, 5, 6, or more) infusions may be employed to deliver a given dose of the engineered IL- 18 polypeptide. In some instances, a dose is administered in a single infusion. In some cases, a dose is divided among multiple infusions. The duration of an infusion (including infusions of a single dose or a portion of a divided dose) will vary and useful infusion durations may include e.g., 15 min. or less, 30 min. or less, 1 hr or less, 2 hrs or less, 3 hrs or less, 4 hrs or less, 6 hrs or less, 8 hrs or less, 10 hrs or less, 12 hrs or less, 24 hrs or less, more than 24 hours, and the like. Useful infusion rates are generally determined based on the desired total time of infusion (e.g., 2 hrs) and the desired volume (e.g., 20 mL) to infuse. Accordingly, useful infusion rates will vary and may include e.g., 1 mL / hr, 5 mL / hr, 10 mL / hr, 15 mL / hr, 20 mL / hr, etc. In some instances, useful resulting infusion rates (with respect to the infused drug substance) may include 0.1 mg / hr or less, 0.1 mg / hr or more, 0.5 mg / hr or less, 0.5 mg / hr or more, 1 mg / hr or less, 1 mg / hr or more, 2 mg / hr or less, 2 mg / hr or more, 3 mg / hr or more, 5 mg / hr or more, 10 mg / hr or more, 0.1 mg / hr to 10mg / hr, 0.5 mg / hr to 10 mg / hr, 1 mg / hr to 10 mg / hr, 1 mg / hr to 5 mg / hr, 2 mg / hr to 5 mg / hr, 1 mg / hr to 3 mg / hr, and the like.

[0149] In some embodiments, the methods of the present disclosure are useful for treating a tumor or cancer that is resistant to immune checkpoint inhibitors (ICIs). Examples of immune checkpoint inhibitors include, but are not limited to: anti-PDl agents such as an anti-PDl antibody (e.g., pembrolizumab, zimberelimab, nivolumab, cemiplimab, dostarlimab), anti-PD- L1 agents such as an anti-PD-Ll antibody (e.g., avelumab, durvalumab, atezolizumab), anti-PD- L2 agents such as an anti-PD-L2 antibody, anti-CTLA4 (e.g., ipilimumab, tremelimumab), anti- TIM3, anti-TIGIT, anti-LAG3 (e.g., relatlimab), anti-B7H3 (e.g., enoblituzumab), anti-B7H4, anti-VISTA, anti-BTLA, anti-CD47, anti- SIRP alpha, anti-CD48, anti-CD155, anti-CD160, anti-TREM2, anti- IDO1, anti-Adenosine 2A receptor, anti -Aryl hydrocarbon receptor, anti- KIR, and anti-LILRB2. Examples of targets of immune checkpoint inhibitors include but are not limited to: PD-L1, PD-L2, PD1, CTLA4, TIM3, TIGIT, LAG3, B7-1, B7H3, B7H4, VISTA, BTLA, CD28, CD47, SIRP alpha, CD48, CD155, CD160, TREM2, IDO1, Adenosine 2A receptor, Aryl hydrocarbon receptor, KIR, LILRB2, and combinations thereof. Other examples of ICIs and targets of ICIs, to which a subject’s cancer may be resistant, are described herein.

[0150] In some instances, subjects treated utilizing the herein described methods are subjects having an absolute neutrophil count (ANC) greater than or equal to 1.0 x 10A9 / L, including greater than or equal to 1.5 x 10A9 / L; platelet count greater than or equal to 75 x 10A9 / L, including platelet count greater than or equal to 100 x 10A9 / L; Hemoglobin (Hgb) greater than or equal to 80 g / L (8.0 g / dL or 4.9 mmol / L), including to 90 g / L (9.0 g / dL or 5.6 mmol / L).

[0151] In some instances, subjects treated utilizing the herein described methods are subjects negative for the following: Human immunodeficiency virus (HIV) antibody (or no history of known HIV infection); Hepatitis B surface antigen (HBsAg) and hepatitis B core antibody or undetectable hepatitis B virus (HBV) deoxyribonucleic acid (DNA); Hepatitis C virus (HCV) antibody or HCV ribonucleic acid (RNA); or any combination thereof.

[0152] In some instances, methods of the present disclosure can include one or more of various assessments. Useful assessments include, e.g., assessing the incidence of DLTs and AEs, changes in clinical laboratory parameters, vital signs, electrocardiogram (ECG) parameters, physical examination findings, the incidence of anti -drug antibodies (AD As) and neutralizing antibodies, and disease assessments. Assessments can be performed at various frequencies including e.g., daily, every other day, twice weekly, weekly, bi-weekly, tri-weekly, monthly, every 6 weeks, every 12 weeks, annually, or at 8- to 12-week intervals.

[0153] In some instances, imaging analysis can be performed, including e.g. computed tomography (CT) or magnetic resonance imaging (MRI) scan of one or more relevant areas,such as e.g., the target lesion, the chest, abdomen, and pelvis. In some instances, imaging can be compared to baseline images, such as e.g., baseline scans, and the methods can include collecting such baseline images. In some instances, assessments can be made through the use of one or more biological samples, such as e.g., blood, plasma, and the like. In some instances, blood and plasma samples can be collected and used for pharmacokinetic (PK) and pharmacodynamic (PD) analysis, as well as for measuring ADA and identifying select biomarkers.

[0154] In some instances, the methods can include one or more additional treatments, which can be routine / scheduled / predetermined or triggered by one or more of the assessments described herein. In some instances, such additional treatments can be performed before administration of an engineered IL- 18, during administration with an engineered IL- 18, or following administration of an engineered IL-18.

[0155] In some instances, a particular therapy can be withheld from the subject prior to, during, and / or following pretreatment with a B-cell depleter. For example, in some instances, treatment with a blood pressure medication can be withheld, e.g., for at least 24 hours before and at least 48 hours after an engineered IL- 18 dose.

[0156] Non-limiting examples of additional treatments can include e.g., administration of: an oral NSAID, e.g., 1-2 hours prior to therapy and continuing for at least 48 hours post-dose (inc., e.g., indomethacin 25 mg 3 times daily or oral ibuprofen 200 to 600 mg every 6 to 8 hours); oral acetaminophen (e.g., 650 mg every 4 to 6 hours as needed for the first 24 hours post-dose or oral acetaminophen 650 mg every 4 to 6 hours beginning 1 hour prior to engineered IL-18 dosing and continuing for at least 48 hours post-dose); IV or oral diphenhydramine (e.g., 50 mg or a comparable dose of a next-generation histamine Hl antagonist 30-60 minutes prior to dosing with engineered IL- 18); up to 3 liters of supplemental fluids in the 24 hours post-dose of engineered IL-18; famotidine (or other H2 blocker, e.g., at 20 mg IV or 20-40 mg orally, 30-60 minutes prior to dosing with engineered IL-18), and any combination thereof. In some instances, steroids can be administered, e.g., as a premedication (e.g., administered at one or more of the premedication timepoints described herein), including e.g., if the subject has hypoadrenalism.

[0157] In some instances, prophylactic antibiotics can be administered, and subjects can receive therapeutic antibiotics for intercurrent infections as needed.

[0158] In some instances, the methods can include hematopoietic supportive therapy, including but not limited to e.g.., granulocyte colony-stimulating factor (G-CSF; e.g., filgrastim, filgrastim-sndz, pegfilgrastim, lenograstim); erythropoietic agents (e.g., erythropoietin or darbepoetin); red blood cell or platelet transfusions; or any combination thereof.

[0159] In some instances, the engineered IL- 18 polypeptide can be administered as a monotherapy. In some instances, the engineered IL- 18 polypeptide can be administered as part of a combination therapy, including e.g., where the combination therapy includes one or more immune checkpoint inhibitors (ICIs).

[0160] Useful ICIs include Zimberelimab, Nivolumab, Cemiplimab, Dostarlimab, Vopratelimab, Spartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, INCMGA00012 , Pidilizumab, MEDI0680, SSI-361, AMP-224, Cetrelimab, Prolgolimab, Retifanlimab, Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab, CA-170, BMS-986189, BMS-936559, Sugemalimab, Adebrelimab, CBT-502, BGB-A333, Pacmilimab, Ipilimumab, Tremelimumab, Relatlimab, Enoblituzumab, the like, and a variant of any of the foregoing.

[0161] In some embodiments, the method comprises administering to a subject, cell, or tissue, an isolated nucleic acid molecule encoding one or more engineered IL-18 polypeptides (e.g., a DR IL-18 polypeptide or a stabilized DR IL-18). In some embodiments, an engineered IL-18 polypeptide is administered to a cell, tissue, organ, system, or subject to treat a disease or disorder, including e.g., as a nucleic acid (e.g., DNA, cDNA, mRNA, etc.) encoding at least one engineered IL- 18 polypeptide.

[0162] Increased IL- 18 signaling, including by using an engineered IL- 18 polypeptide, can be assessed using a wide variety of methods, including those disclosed herein, as well as methods known in the art or to be developed in the future. That is, the skilled artisan would appreciate, based upon the disclosure provided herein, that increasing the level or activity of IL- 18 signaling can be readily assessed using methods that assess the level of a nucleic acid (e.g., mRNA) encoding IL- 18 or an engineered IL- 18 polypeptide or fragment thereof, the level of IL- 18 or an engineered IL-18 polypeptide or fragment polypeptide, and / or the level of IL-18 or an engineered IL-18 polypeptide or fragment activity in a biological sample obtained from a subject.COMPOSITIONS, AGENTS & DEVICES

[0163] Compositions for use the herein described methods include engineered IL- 18 polypeptide compositions, including e.g., DR IL-18 polypeptide compositions, D2D IL-18 polypeptide compositions, and the like. Compositions for use the herein described methods can include essentially one therapeutic or active agent or multiple different therapeutics or active agents. For example, useful compositions can include engineered IL- 18 polypeptide compositions useful in monotherapy, separate engineered IL- 18 polypeptide and ICI compositions useful in combination therapy, or combination compositions (e.g., including an engineered IL-18 polypeptide and ICI in a single composition) useful in combination therapy.

[0164] In some instances, compositions of the present disclosure are therapeutic compositions useful in treating a disease, including e.g., cancer (including e.g., solid tumor cancers), infections (including e.g., viral infections, bacterial infections, parasitic infections, etc.), diseases related to decreased or repressed immunity (including e.g., neutropenia, e.g., as a result of chemotherapy), genetic and metabolic diseases, and the like.

[0165] Useful engineered IL-18 polypeptide compositions can include essentially any useful engineered IL-18 polypeptide, including but not limited to e.g., an engineered IL-18 polypeptide comprising any of the amino acid sequences of SEQ ID NOs. :2-97. In some instances, an engineered IL-18 polypeptide composition includes an engineered IL-18 polypeptide comprising an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% with at least one of SEQ ID NOs: 1 -63.

[0166] In some instances, an engineered IL-18 polypeptide composition includes a stabilized engineered IL- 18 polypeptide. A stabilized IL- 18 polypeptide can include mutations of two cysteine residues (e.g., C38 and C68) relative to human WT IL- 18 (SEQ ID NO:01). In some cases, the mutation is a C to S substitution and as such the stabilized IL- 18 polypeptide can in some cases comprise the mutations C38S and C68S. In some cases a stabilized IL-18 polypeptide comprises the mutation pair C38S / C68S, C38S / C68G, C38S / C68A, C38S / C68V, C38S / C68D, C38S / C68E, or C38S / C68N (e.g., in some cases C38S / C68G, C38S / C68A, C38S / C68V, C38S / C68D, C38S / C68E, or C38S / C68N; in some cases C38S / C68S, C38S / C68G, C38S / C68A, C38S / C68D, or C38S / C68N; and in some cases C38S / C68G, C38S / C68A, C38S / C68D, or C38S / C68N).

[0167] In some cases, a stabilized IL- 18 polypeptide is a stabilized IL- 18 variant polypeptide (e.g., a stabilized "decoy resistant" (DR) IL-18 variant or a stabilized "decoy -to-the-decoy" (D2D) IL-18 variant), i.e., an engineered IL-18 polypeptide that additionally includes stabilizing mutations, e.g., mutations at positions C38 and C68 relative to human wild type IL-18.

[0168] In some embodiments, the compositions and methods of the disclosure include an engineered IL-18 polypeptide that is mutated relative to wild type IL-18 such that it can bind to one of IL-18's natural binding partners, but not to the other (or has reduced binding to the other).

[0169] For example, wild type IL- 18 binds to both IL-18R (to signal through the receptor) and IL-18BP (which inhibits IL- 18 by preventing it from binding to IL-18R). In some cases, an IL- 18 variant polypeptide is a "decoy resistant IL-18" ("DR IL-18") variant; such variants bind to IL-18R but have reduced binding to (and in some cases do not bind to) IL-18BP. In other cases, an IL-18 variant polypeptide discussed herein is a "decoy -to-the-decoy IL-18" ("D2D IL- 18") variant; such variants bind to IL-18BP but have reduced binding to (and in some cases do not bind to) IL-18R. An IL- 18 variant polypeptide can show comparable binding affinity to IL-18Ras wild type IL-18, while also exhibiting decreased binding affinity to IL-18BP compared to wild type IL-18.

[0170] DR-IL-18 variants and D2D IL- 18 variants can both, additionally, be stabilized variants, through the presence of one or more stabilizing mutations. DR-IL-18 variants and D2D IL-18 variants, including in stabilized and non-stabilized forms, that are useful as engineered IL- 18 polypeptides that are described herein and also in PCT Pub. Nos. W02019 / 051015 and WO2022 / 094473, the disclosures of which are incorporated herein by reference in their entirety.

[0171] In some instances, a useful engineered IL- 18 polypeptide comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with at least one of SEQ ID NOs:l, 7, 17-22, 55-57, 62 and 63. In some instances, a useful engineered IL-18 polypeptide comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, or 15 mutated residues relative to SEQ ID NO: 1. In some instances, a useful engineered IL-18 polypeptide comprises up to 15, 14, 13, 12, 11, or 10 mutated residues relative to SEQ ID NO: 1. In some instances, a useful engineered IL-18 polypeptide comprises from 1 to 20, from 2 to 20, from 3 to 20, from 4 to 20, from 5 to 20, from 6 to 20, from 7 to 20, from 8 to 20, from 9 to 20, from 10 to 20, from 11 to 20, from 12 to 20, from 13 to 20, from 14 to 20, from 15 to 20, from 1 to 18, from 2 to 18, from 3 to 18, from 4 to 18, from 5 to 18, from 6 to 18, from 7 to 18, from 8 to 18, from 9 to 18, from 10 to 18, from 11 to 18, from 12 to 18, from13 to 18, from 14 to 18, from 15 to 18, from 1 to 16, from 2 to 16, from 3 to 16, from 4 to 16, from 5 to 16, from 6 to 16, from 7 to 16, from 8 to 16, from 9 to 16, from 10 to 16, from 11 to16, from 12 to 16, from 13 to 16, from 14 to 16, from 1 to 14, from 2 to 14, from 3 to 14, from 4 to 14, from 5 to 14, from 6 to 14, from 7 to 14, from 8 to 14, from 9 to 14, from 10 to 14, from11 to 14, from 12 to 14, from 1 to 12, from 2 to 12, from 3 to 12, from 4 to 12, from 5 to 12, from 6 to 12, from 7 to 12, from 8 to 12, from 9 to 12, from 10 to 12, from 1 to 10, from 2 to 10, from 3 to 10, from 4 to 10, from 5 to 10, from 6 to 10, from 7 to 10, or from 8 to 10 mutated residues relative to SEQ ID NO: 1.

[0172] In some instances, a useful engineered IL- 18 polypeptide comprises an amino acid sequence of any one of SEQ ID NOs:7, 17-22, 55-57, 62 and 63 with 0, 1, 2, 3, 4, 5, or 6 additional mutated residues relative to SEQ ID NO: 1.

[0173] In some instances, a useful engineered IL-18 polypeptide comprises one or more mutations at positions Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DI 10, N111, Ml 13, V153, and N155 relative to the WT IL-18 amino acid sequence set forth in SEQ ID NO: 1, including e.g., at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least, 8, at least 9, at least 10, at least 11, or at least 12 of the listed mutated positions. Accordingly, useful engineered IL-18 polypeptides can have one or any combinationof mutations at the listed positions, with or without additional mutated residues at other positions. For example, useful polypeptides can have a mutation at position Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DUO, Nl l l, Ml 13, V153, or N155 only; mutations at all positions Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DI 10, Nl l l, Ml 13, V153, and N155; or any combinations having more than one but less than all (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or 19) mutations at these positions.

[0174] The engineered IL-18 polypeptide comprises at least 12 mutations, at least 11 mutations, at least 10 mutations, at least 9 mutations, at least 8 mutations, at least 7 mutations, at least 6 mutations, at least 5 mutations, at least 4 mutations, at least 3 mutations, at least 2 mutations, or at least 1 mutation relative to the wild-type IL-18 amino acid sequence set forth in SEQ ID NO: 1, optionally wherein the at least 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 mutations are at positions selected from Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DUO, Nl l l, Ml 13, V153, and N155.

[0175] In some instances, the engineered IL-18 polypeptide is a DR IL-18 polypeptide that comprises 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 7, 17- 22, 55-57, 62 and 63.

[0176] In some instances, the DR- 18 polypeptide has less than 100% and at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 88%, at least 87%, at least 86%, or at least 85% sequence identity to WT human IL- 18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO: 102), wherein the X at pos. 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. I l l is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H.

[0177] In some instances, the DR-18 polypeptide comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, atleast 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 substitutions relative to WT human IL-18 (SEQ ID NO: 1) and comprises the sequence XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO: 102), wherein the X at pos. 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. I l l is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H.

[0178] In some instances, the DR-18 polypeptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 substitutions relative to WT human IL-18 (SEQ ID NO: 1) and comprises the sequence XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO: 102), wherein the X at pos. 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. I l l is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H.

[0179] In some instances, the DR- 18 polypeptide comprises one or more substitutions relative to WT human IL-18 (SEQ ID NO: 1), including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more substitutions selected from: Y1H, Y1R, L5H, L5I, L5Y, K8Q, K8R, M51T, M51K, M51D, M51N, M51E, M51R, K53R, K53G, K53S, K53T, S55K, S55R, Q56E, Q56A, Q56R, Q56V, Q56G, Q56K, Q56L, P57L, P57G, P57A, P57K, G59T, G59A, M60K, M60Q, M60R, M60L, E77D, Q103E, Q103K, Q103P, Q103A, Q103R, S105R, S105D, S105K, S105N, S105A, D110H, D110K, DI 10N, DI 10Q, DI 10E, DI 10S, DI 10G, N111H, N111 Y, N11 ID, N111R, N11 IS, N111G, Ml 13V, M113R, M113T, M113K, V153I, V153T, V153A, N155K, and N155H.

[0180] In some instances, the DR-18 polypeptide comprises one or more substitutions relative to WT human IL-18 (SEQ ID NO: 1) such that, e.g., the amino acid at pos. 1 is not Y and, e.g., may be R or H; the amino acid at pos. 5 is not L and, e.g., may be H, I or Y; the amino acid at pos. 8 is not K and, e.g., may be Q or R; the amino acid at pos. 38 is not C and, e.g., may be S; the amino acid at pos. 51 is not M and, e.g., may be T, K, D, N, E or R; the amino acid at pos. 53 is not K and, e.g., may be R, G, S or T; the amino acid at pos. 55 is not S and, e.g., may be K or R; the amino acid at pos. 56 is not Q and, e.g., may be E, A, R, V, G, K, L or R; the amino acid at pos. 57 is not P and, e.g., may be L, G, A or K; the amino acid at pos. 59 is not G and, e.g., may be A or T ; the amino acid at pos. 60 is not M and, e.g., may be K, Q, R or L; the amino acid at pos. 68 is not C and, e.g., may be S, G, A, V, D, E or N ; the amino acid at pos. 76 is not C and, e.g., may be S; the amino acid at pos. 77 is not E and, e.g., may be D; the amino acid at pos. 103 is not Q and, e.g., may be E, K, P, A or R; the amino acid at pos. 105 is not S and, e.g., may be D, N, R, K or A; the amino acid at pos. 110 is not D and, e.g., may be K, H, N, Q, E, S or G; the amino acid at pos. I l l is not N and, e.g., may be H, Y, D, R, S or G; the amino acid at pos. 113 is not M and, e.g., may be V, R, T or K; the amino acid at pos. 127 is not C and, e.g., may be S; the amino acid at pos. 153 is not V and, e.g., may be I, T or A; and / or the amino acid at pos. 155 is not N and, e.g., may be K or H.

[0181] In some instances, the DR-18 polypeptide comprises substitution mutations, relative to WT human IL-18 as set forth in SEQ ID NO: 1, at positions: (i) M51, M60, S 105, DI 10, and Nl l l; (ii) M51, S55, G59, M60, S105, DI 10, N111, and V153; (iii) Yl, M51, M60, S105, DUO, and Nl l l; (iv) Yl, M51, K53, M60, S105, DUO, and Nl l l; (v) K8, M51, S55, G59, M60, S105, DUO, and N155; (vi) K8, M51, S55, G59, M60, S105, DI 10, N111, and V153; (vii) L5, M51, K53, M60, S105, DUO, and V153; (viii) L5, M51, S55, G59, M60, S105, DUO, Nl l l, and N155; (ix) L5, M51, S55, M60, Q103, S105, DI 10, N111, and V153; (x) L5, M51, S55, M60, S105, DI 10, Nl l l, V153, and N155; (xi) L5, M51, S55, G59, M60, S105, DUO, N111, V153, and N155; (xii) L5, K8, M51, S55, M60, S105, N111, V153, and N155; (xiii) L5, K8, M51, K53, M60, S105, DUO, Nl l l, and N155; (xiv) Yl, L5, M51, K53, M60, S105, DUO, and N155; (xv) Y1, M51, K53, G59, M60, S105, DI 10, N111, V153, and N155; (xvi) Y1, K8, M51, K53, M60, Q103, S105, DUO, Nl l l, and N155; (xvii) Yl, K8, M51, M60, S105, DUO, and Nl l l; (xviii) Yl, L5, M51, K53, M60, Q103, S105, DUO, and Nl l l; (xix) Yl, K8, M51, K53, G59, M60, Q103, S105, DI 10, N111, V153, and N155; (xx) Yl, K8, M51, K53, G59, M60, S105, DUO, Nl l l, and N155; (xxi) Yl, K8, M51, G59, M60, Q103, S105, DI 10, N111, V153, and N155; (xxii) Yl, L5, M51, G59, M60, E77, S105, DUO, and Nl l l; (xxiii) M51, Q56, P57, M60, Q103, S105, DI 10, N111, and Ml 13; (xxiv) M51, Q56, P57, M60, Q103, S105, DUO, and Ml 13; (xxv) M51, K53, Q56, P57, M60, DUO, and Nl l l; (xxvi) M51, K53, Q56,P57, M60, Q103, S105, DI 10, N111, and Ml 13; (xxvii) M51, K53, Q56, M60, Q103, S105, DI 10, N111, and Ml 13; (xxviii) M51, K53, Q56, P57, Q103, S105, DI 10, N111, and Ml 13; (xxix) M51, K53, Q56, P57, M60, S105, DUO, and Nl l l; (xxx) M51, K53, Q56, P57, M60, Q103, DI 10, N111, and Ml 13; (xxxi) M51, Q56, P57, M60, Q103, DI 10, N111, and Ml 13; or (xxxii) M51, K53, Q56, S105, DUO, and Nl l l.

[0182] In some instances, the DR-18 polypeptide comprises substitution, relative to WT human IL- 18 as set forth in SEQ ID NO: 1, of: (i) M51T, M60K, S105D, DI 10K, and N111H; (ii) M51T, S55K, G59A, M60K, S105D, DI 10K, N111H, and V153I; (iii) Y1R, M51T, M60K, S105D, D110K, and Nl l lH; (iv) Y1R, M51T, K53R, M60K, S105N, D110K, and Nl l lY; (v) K8Q, M51T, S55K, G59T, M60K, S105R, D110H, and N155K; (vi) K8R, M51K, S55K, G59A, M60Q, S105D, D110K, Nl l lH, and V153I; (vii) K8R, M51D, S55K, G59A, M60X, S105D, D110K, N111H, and V153I; (viii) L5H, M51T, K53R, M60K, S105D, DI 10N, and V153T; (ix) L5I, M51K, S55K, G59A, M60Q, S105K, DI 10Q, N111H, and N155K; (x) L5I, M51T, S55R, M60K, Q103E, S105D, DI 10H, N111H, and V153I; (xi) L5I, M51T, S55K, M60K, S105D, D110K, N111H, V153T, and N155H; (xii) L5I, M51T, S55K, G59A, M60K, S105R, D110H, N111H, V153I, and N155K; (xiii) L5I, K8R, M51T, S55K, M60K, S105D, N111Y, V153I, and N155K; (xiv) L5Y, K8R, M51T, K53R, M60K, S105D, D110E, N111H, and N155K; (xv) Y1H, L5Y, M51T, K53R, M60K, S105D, D110H, and N155K; (xvi) Y1R, M51T, K53R, G59A, M60K, S105D, D110Q, N111H, V153A, and N155K; (xvii) Y1R, K8R, M51D, K53R, M60R, Q103K, S105N, D110K, Nl l lY, and N155H; (xviii) Y1R, K8R, M51N, K53R, M60Q, Q103K, S105R, D110N, Nl l lH, and N155K; (xix) Y1R, K8R, M51T, M60K, S105D, D110K, and N111H; (xx) Y1R, L5H, M51T, K53R, M60K, Q103E, S105N, D110K, and Nl l lY; (xxi) Y1R, K8R, M51T, K53R, G59A, M60K, Q103E, S105D, D110Q, N111H, V153I, and N155X; (xxii) Y1R, K8R, M51T, K53R, G59T, M60K, S105N, D110H, Ni l ID, and N155H; (xxiii) Y1R, K8R, M51T, G59A, M60K, Q103E, S105D, D110Q, N111H, V153I, and N155K; (xxiv) Y1R, L5Y, M51T, G59T, M60K, E77D, S105D, D110K, and Nl l lH; (xxv) Y1R, K8R, M51T, K53R, G59T, M60K, S105K, DI 10N, N111H, and N155K; (xxvi) M51E, Q56E, P57L, M60R, Q103P, S105A, DI 10N, Ni l 1R, and Ml 13V; (xxvii) M51K, Q56A, P57G, M60L, Q103E, S105D, DUOS, and Ml 13V; (xxviii) M51K, K53G, Q56A, P57A, M60L, D110K, and Nl l lR; (xxix) M51K, K53G, Q56R, P57G, M60L, Q103E, S105D, DI 10N, N11 IS, and M113R; (xxx) M51K, K53G, Q56V, M60L, Q103A, S105A, DI IOS, N111R, and M113T; (xxxi) M51K, K53S, Q56G, P57A, M60L, Q103A, S105A, DI 10G, N111R, and M113T; (xxxii) M51K, K53S, Q56K, P57A, QI 03 A, S105D, DI IOS, N11 IS, and M113R; (xxxiii) M51K, K53S, Q56L, P57A, M60L, S105D, DUOS, and Nl l lR; (xxxiv) M51K, K53S, Q56R, P57A, M60L, S105N, DI 10G, and N111R; (xxxv) M51K, K53S, Q56R, P57A, M60L, QI 03 A, DI 10G, N111R, andM113T; (xxxvi) M51K, K53S, Q56R, P57A, M60L, Q103A, S105D, DUOS, Ni l 1G, and M113R; (xxxvii) M51K, K53T, Q56R, M60L, Q1O3E, S1O5D, DUOS, N111S, and M113K; (xxxviii) M51K, K53T, Q56R, P57A, Q1O3E, S1O5D, DI ION, N11 ID, and M113R; (xxxix) M51R, Q56G, P57K, M60L, Q1O3R, DI IOS, N111R, and Ml 13 V; (xl) M51K, K53G, Q56G, P57A, M60L, Q1O3E, S1O5D, DI IOS, N111G, and Ml 13V; or (xli) M51K, K53G, Q56R, SI 05 A, DI ION, and Ni l 1R.

[0183] Useful IL-18 variants, as well as useful mutations and substitutions thereof, include but are not limited to those variants, mutations and substitutions described in W02002101049 (such as e.g., variants having mutations at positions E42 and / or K89 and substitutions E42A and K89A (numbering according to the pre-processed IL- 18 polypeptide)), US20080206189 (such as e.g., variants having mutations at one or more of C38, C68, C76, N78, E121, C127, L144, and D157 and substitutions, individually or in combination, of C38S, C68D, C68S, N78C, E121C, L144C, and D157C), WO2023118497 (such as e.g., variants having mutations at one or more of Yl, G3, S10, C38, M51, K53, S55, P57, M60, C68, Q103, Ml 13, C127, and N155, and substitutions, individually or in combination, of Y1A, G3Y, S10R, S10K, C38S, M51A, M51Y, M51Q, K53L, K53H, K53A, S55L, S55Y, P57T, M60A, M60E, M60H, C68S, Q103S, M113A, M113E, C127S, and N155Y (inc. those at pos. K53 other than K53R, K53G, K53S, or K53T), WO2023114829 (such as e.g., variants having mutations at one or more of Y37, S43, L45, S46, V47, N50, F57, C74, T81, 185, S86, D90, S91, A97, V98, T99, S101, C104, 1107, T109, Cl 12, N123, P124, Q139, R140, Q150, A162, C163, L174, 1185, V189, Q190, and E192, and substitutions, individually or in combination, of Y37C, S43C, L45C, S46C, V47C, N50C, F57C, C74S, T81C, I85C, S86C, D90C, S91C, A97C, V98C, T99C, S101C, C104S, I107C, T109C, C112S, N123C, P124C, Q139C, R140C, Q150C, A162C, C163S, L174C, I185C, V189C, Q190C, E192C, (numbering according to the pre-processed IL- 18 polypeptide)), WO2023056193 (such as e.g., variants having mutations at one or more of M51, K53, Q56, P57 and M60, and substitutions, individually or in combination, of M51A, M51S, M51G, K53G, K53A, K53S, Q56R, Q56K, Q56H, P57A, P57S, P57G, M60R, M60K, and M60H (and useful sequences encoding such variants, such as e.g., SEQ ID NOs: 7 and 8 of WO2023056193)), US20230146665 (such as e.g., variants having mutations at one or more of Yl, E6, S7, K8, S10, V11, N14, L15, D17, Q18, D23, R27, P28, L29, E31, M33, T34, D35, S36, D37, C38, R39, D40, N41, R44, 146, 149, S50, M51, K53, D54, S55, Q56, P57, M60, A61, V62, T63, S65, K67, C68, E69, 171, C76, E77, 180, 181, N87, P88, D90, K93, T95, K96, S97, Q103, H109, DUO, Nl l l, M113, SI 19, A126, C127, D132, L136, L138, K139, E141, L144, D146, R147, 1149, M150, N155, E156, and D157, and substitutions, individually or in combination, of Y1F, Y1H, E6A, E6Q, S7C, S7P, K8E, K8Q, K8Y, S10C, VI II, N14C, N14W, L15C, DUN, Q18L, D23N,D23S, R27Q, P28C, L29V, E31Q, M33C, T34P, D35N, D35E, S36D, S36N, D37N, C38S, C38Q, C38R, C38E, C38L, C38I, C38V, C38K, C38D, R39S, R39T, D40N, N41Q, R44Q, I46V, I49C, S50C, S50Y, M51I, M51K, M51Q, M51R, M51L, M51H, M51F, M51Y, K53A, K53D, K53E, K53G, K53H, K53I, K53L, K53M, K53N, K53Q, K53R, K53S, K53T, K53V, K53Y, K53F, D54C, S55N, S55Q, S55D, S55E, S55T, Q56I, Q56L, P57A, P57E, P57T, P57V, P57Q, P57D, P57Y, P57N, M60I, M60L, M60K, M60Y, M60F, A61C, V62C, T63C, S65C, K67Q, C68S, C68I, C68F, C68Y, C68D, C68N, C68E, C68Q, C68K, E69K, I71M, C76S, C76E, C76K, E77K, 180T, 18 IL, 18 IV, N87S, P88C, D90E, K93D, K93N, T95E, K96G, K96Q, S97N, Q103C, Q103E, Q103I, Q103L, H109W, H109Y, DI ION, DI 1OQ, DI 1OR, N11 ID, N11 IQ, N11 IS, N11 IT, N11 IE, Ml 131, SI 19L, A126C, C127S, C127W, C127Y, C127F, C127D, C127E, C127K, D132Q, D132E, L136C, L138C, K139C, E141K, E141Q, L144N, D146F, D146L, D146Y, R147C, R147K, I149V, M150F, M150T, N155C, E156Q, D157A, D157S, and D157N, as well as deletion mutants described therein), US20220056091 (such as e.g., variants having mutations at one or more of Y01, F02, E06, M33, C38, M51, K53, D54, S55, M60, T63, C68, E69, K70, C76, M86, Ml 13, C127, and M150, and substitutions, individually or in combination, of Y01G, F02A, E06K, C38A, C38S, K53A, D54A, S55A, T63A, C68A, C68S, E69C, K70C, C76A, C76S, C127A, and C127S), US20230357342 (such as e.g., variants having mutations at one or more of Yl, F2, E6, K8, S10, VI 1, D17, E31, T34, D35, S36, D37, C38, D40, N41, 149, M51, K53, D54, S55, T63, C68, E69, K70, C76, Q103, S105, G108, H109, DUO, C127, D132, and V153, and substitutions, individually or in combination, of YIM, Y1G, F2A, E6K, E6R, K8E, K8L, K8R, S10T, VI II, D17N, E31A, T34A, D35A, S36A, D37A, C38A, C38Q, C38S, D40A, N41A, I49E, I49M, I49R, M51G, K53A, D54A, S55A, S55H, S55R, S55T, T63A, C68A, C68S, E69C, K70C, C76A, C76S, Q103E, Q103K, Q103R, S105I, S105K, G108A, H109A, D110A, C127A, C127S, D132A, V153E, V153R, and V153Y), and WO2022172944 (such as e.g., variants having mutations at one or more of C38, C68, C76, C127, G3, A6, S7, T34, S50, M51, S72, KI 12, KI 19, and G145, and substitutions, individually or in combination, of C38S, C68S, C76S, C127S, G3Y, G3L, A6W, S7C, T34P, C38M, S50C, M51Y, S72Y, S72F, S72M, S72L, S72W, K112W, KI 19V, and G145N).

[0184] Useful IL-18 variants, as well as useful mutations and substitutions thereof, further include but are not limited to those variants, mutations and substitutions described in PCT Pub. Nos. W02024044780, W02004091517, WO2024102587, WO2022038417, WO2023161856, WO2024051728, W02003031569, and WO2024102693; and US Pat. Pub. No.US20040023336; and the like; the disclosures of which are incorporated by reference in their entirety.

[0185] Useful engineered IL-18 polypeptides, including useful DR IL-18 polypeptides and IL- 18 variants, include those consisting essentially of the engineered, DR, or variant polypeptide itself, i.e., without any additional polypeptides fused and / or conjugated to the engineered, DR, or variant polypeptide. Useful engineered IL-18 polypeptides include, in some instances, engineered polypeptides that are fused and / or conjugated to one or more other polypeptides, such as but not limited to e.g., immunoglobulin polypeptides or fragments or portions thereof (e.g., antibodies, antigen binding domains, variable domains, scFv domains, Fc domains, and the like), albumin polypeptides, cytokines or fragments thereof, cleavable polypeptides, masking polypeptides, and the like. Accordingly, in some instances, the engineered IL-18 polypeptide employed in the method may be a fusion protein (or conjugation of two or more polypeptide chains) comprising at least one non-IL-18 polypeptide domain and at least one engineered IL- 18 polypeptide domain, such as e.g., a DR IL- 18 polypeptide domain. In some instances, the engineered IL-18 polypeptide is not a fusion protein and is not a conjugated protein.

[0186] In some instances, the engineered IL- 18 polypeptide employed in the method is selected from SEQ ID NOs: 7, 17-22, 55-57, and 62. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 7. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 17. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 18. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 19. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 20. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 21. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 22. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 55. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 56. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 57. In some instances, the engineered IL-18 polypeptide employed in the method is SEQ ID NO: 62.

[0187] In some instances, the engineered IL- 18 polypeptide is a D2D IL- 18 polypeptide that comprises 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 64- 97. In some instances, the engineered IL-18 polypeptide is a D2D IL-18 polypeptide that comprises at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 64-97.

[0188] In some embodiments, the engineered IL-18 polypeptide is a monomer. In some embodiments, the monomer is not in a protein complex. In some embodiments, the monomer isfunctional as a single polypeptide. In some embodiments, the engineered IL- 18 polypeptide is not glycosylated. In some embodiments, the engineered IL-18 polypeptide is glycosylated. In some embodiments, the engineered IL- 18 polypeptide is partially glycosylated. In some embodiments, the engineered IL-18 polypeptide is at least about 50% glycosylated. In some embodiments, the engineered IL-18 polypeptide is at most about 50% glycosylated.

[0189] In some embodiments, the engineered IL- 18 polypeptide occupies between 10% to about 50% of the IL- 18 receptor. In some embodiments, the engineered IL- 18 polypeptide occupies at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more of the IL- 18 receptor. In some embodiments, the engineered IL-18 polypeptide occupies at most about 50%, at most about 45%, at most about 40%, at most about 35%, at most about 30%, at most about 25%, at most about 20%, at most about 15%, at most about 10%, or less of the IL- 18 receptor. In some embodiments, the engineered IL- 18 polypeptide occupies about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of the IL- 18 receptor.

[0190] In some embodiments, e.g., in combination therapy with an engineered IL- 18 polypeptide, useful compositions also include immune checkpoint inhibitor (ICI) compositions. In some embodiments, a useful ICI is a binding antagonist. In some embodiments, the ICI is an immunoadhesin. In some embodiments, the ICI is an antibody or antigen binding fragment thereof. In some embodiments, the ICI is a monoclonal antibody or antigen binding fragment thereof.

[0191] Non-limiting examples of useful PD-1 / PD-L1 antagonist ICIs include: Pembrolizumab, Zimberelimab, Nivolumab, Cemiplimab, Dostarlimab, Vopratelimab, Spartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, INCMGA00012, Pidilizumab, MEDI0680, SSI-361, AMP -224, Cetrelimab, Prolgolimab, Retifanlima, Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab, CA-170, BMS-986189, BMS-936559, Sugemalimab, Adebrelimab, CBT-502, BGB-A333, and Pacmilimab. A non-limiting example of a PD-L2 antagonist ICI is CA-170.

[0192] As described above, methods of the present disclosure can make use of one or more CRS prophylactic agents. One or more CRS prophylactic agents can be present in a CRS prophylactic composition. Useful CRS prophylactic agents that can be present in such a composition, include e.g., NSAIDs, analgesics, acetaminophen, antihistamines, histamine Hl antagonists, diphenhydramine, H2 blockers (i.e., histamine H2 antagonists), famotidine, cytokine inhibitors, supplemental fluids and steroids.

[0193] Non-limiting examples of NSAIDs include ibuprofen, naproxen, diclofenac, diflunisal, fenoprofen, flurbiprofen, ketoprofen, meloxicam, nabumetone, oxaproin, piroxicam, etodolac, indomethacin, ketorolac, nabumetone, sulindac, tolmetin, celecoxib, rofecoxib, valdecoxib, mefenamic acid, etoricoxib, indomethacin, and aspirin. In some embodiments, the amount of the NSAID administered is from about 25 mg to about 60 mg, including e.g., from about 25 mg to about 60 mg of diclofenac. In some embodiments, the amount of the NSAID administered is from about 200 mg to about 600 mg, including e.g., from about 200 mg to about 600 mg of ibuprofen.

[0194] Non-limiting examples of analgesics include acetaminophen, aspirin, caffeine, butalbital, isometheptene mucate, magnesium salicylate, codeine, dihydrocodeine, hydrocodone, oxycodone, tramadol, co-codamol, co-codaprin, co-dydramol, alfentanil, fentanyl, hydromorphone, methadone, morphine, meperidine, and oxymorphone. In some embodiments, the amount of the analgesic administered is at least about 500 mg or at least about 600 mg, including e.g., at least about 500 mg or at least about 600 mg of acetaminophen.

[0195] Non-limiting examples of histamine Hl antagonists include brompheniramine, chlorpheniramine, clemastine, cypropheptadine, dexchlorpheniramine dimenhydrinate, diphenhydramine, doxylamine, hydroxyzine, phenindamine, azelastine, loratadine, cetirizine, desloratadine, and fexofenadine. In some embodiments, the amount of the Hl antagonist administered is at least about 20 mg or at least about 40 mg, including e.g., at least about 20 mg or at least about 40 mg of diphenhydramine. Non-limiting examples of histamine H2 antagonists include cimetidine, famotidine, nizatidine, and ranitidine. In some embodiments, the amount of the H2 antagonist administered is at least about 20 mg or at least about 40 mg, including e.g., at least about 20 mg or at least about 40 mg of famotidine.

[0196] Cytokine inhibitors include antagonists, soluble receptors, cytokine-binding proteins, and cytokines that block other cytokines. Cytokine inhibitors may target (i.e., antagonize) cytokine receptors. Non-limiting examples of cytokine receptors include TNFa, TNFP, IFNa, IFNP, IFNy, IL-6, and IL-12. For example, in some instances, useful cytokine inhibitors include IL-6 antagonists, such as e.g., IL-6 antagonist antibodies (e.g., tocilizumab), IL-6 small molecule antagonists, and the like. In some embodiments, the amount of the cytokine inhibitor administered is at least about 500 mg, including, e.g., at least about 500 mg of an IL-6 antagonistic antibody, such as e.g., tocilizumab.

[0197] Non-limiting examples of supplemental fluids include normal saline solution, lactated ringer’s solution, dextrose in water solution, sodium chloride solution, and any combinations thereof. In some embodiments, at most about 3 liters of the supplemental fluid is administered.

[0198] Non-limiting examples of steroids include cortisone, hydrocortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, and betamethasone. In some embodiments, prednisone can be used. In some embodiments, the steroid is administered at at most about 10 mg / day, including e.g., at most about 10 mg / day of prednisone.

[0199] Useful formulations of the herein described compositions can vary. In some instances, useful compositions can be formulated as a sterile solution for injection to be administered by SC injection or IV infusion. In some instances, useful composition can be formulated as a liquid formulation in single-use vials. For example, in some instances, an engineered IL- 18 composition can be formulated as a liquid formulation in single-use vials at a concentration of 30 mg / mL, including e.g., where the vial contains at least about 0.5 mL, at least about 1 mL, at least about 1.5 mL, or at least about 2 mL.

[0200] In some embodiments, the composition further comprises a buffering agent. In some embodiments, the buffering agent is selected from the group consisting of: L-Histidine, L- Histidine Hydrochloride, Phosphate, Tris, Citrate, Glycine, Diethanolamine (DEA), and any combination thereof. In some embodiments, the buffering agent is L-Histidine. In some embodiments, the embodiments, the buffering agent is L-Histidine Hydrochloride. In some embodiments, the buffering agent is L-Histidine and L-Histidine Hydrochloride.

[0201] In some embodiments, the composition further comprises a stabilizer. In some embodiments, the stabilizer is selected from the group consisting of: glycerol, sucrose, glucose, therhalose, ethylene glycol, glycerol, erythritol, threitol, arabitol, ethylenediaminetetraacetic acid (EDTA), disodium salt of EDTA, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, volemitol, isomalt, maltitol, lactitol, maltotriitol, maltotetritol, polyglycitol, and any combination thereof. In some embodiments, the stabilizer is sucrose. In some embodiments, the stabilizer is EDTA. In some embodiments, the stabilizer is a disodium salt of EDTA. In some embodiment, the stabilizer comprises sucrose and EDTA. In some embodiments, the stabilizer comprises sucrose and the disodium salt of EDTA. In some embodiments, the stabilizer comprises EDTA and the disodium salt of EDTA. In some embodiments, the stabilizer comprises sucrose, EDTA, and disodium salt of EDTA.

[0202] In some embodiments, the composition further comprises a surfactant. In some embodiments, the surfactant is an anionic surfactant. Non-limiting examples of anionic surfactants include docusate, perfluorooctanesulfonate, perfluorobutanesulfone, alkyl-aryl ester phosphate, and alkyl ether phosphate. In some embodiments, the surfactant is a cationic surfactant. Non-limiting examples of cationic surfactants include cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, dimethyldioctadecylammonium chloride, and dioctadecyldimethylammonium bromide. In some embodiments, the surfactant is a non-ionicsurfactant. Non-limiting examples of non-ionic surfactants include octaethylene glycol monododecyl ether, Triton X-100, polyethoxylated tallow amine, cocamide monoethanolamine, poloxamer, glycerol monostearate, glycerol monolaurate, sorbitan monolaurate, sorbitan monostearate, sorbitan tristearate, Tween 20, Tween 80, decyl glucoside, octyl glucoside, and polysorbate 80. In some embodiments, the surfactant is polysorbate 80.

[0203] In some embodiments, the composition further comprises an anti-oxidation agent. In some embodiments, the anti-oxidation agent is selected from the group consisting of ascorbic acid, glutathione, lipoic acid, uric acid, carotene, a-tocopherol, ubiquinol, and L-Methionine. In some embodiments, the anti-oxidation agent is L-Methionine.

[0204] In some embodiments, the composition further comprises a solvent. Non-limiting examples of solvents include water, ethanol, methanol, acetone, tetracholoroethylene, toluene, methyl acetate, and ethyl acetate. The composition can be present in any suitable container.

[0205] In some embodiments, the composition can be configured for IV use, including e.g., where the formulation is configured for IV use and / or the composition / formulation is present in a suitable container for IV delivery. Suitable containers can include vials, tubes, bottles, bags, syringes, autoinjectors, and the like, and can be configured for direct administration or mixing with other agents delivered concurrently. For example, in some instances, the composition can be present, prepared for IV injection, in a suitable syringe for direct or indirect delivery to the subject. In some instances, the composition can be present in container, such as a vial, with a puncturable closure allowing for easy sterile transfer of the composition into a syringe or other suitable device for injection directly into the subject or a secondary delivery device (e.g., an IV port, reconstitution IV bag (e.g., Mini-Bag or Mini-Bag Plus Container System) or other component of an IV apparatus). In some instances, the composition can be present in an IV bag container.

[0206] Suitable devices can be calibrated in milliliters and scaled from the tip to the fill line. Calibrated space can be built into the device to permit addition of other drugs. Containers can contain suitable labels, including e.g., labels indicating that the contents deliver X mg (or g or mcg) in Y mL of solution.

[0207] Suitable devices can include a needle of an appropriate size, which can be indicated on the container. In some instances, the needle can be an integral part of the device. In some instances, the device can include a suitable plunger, which can e.g., be configured to be mechanically stable in the barrel of a syringe. In some instances, the device can be of such a design that it is patient ready and assembly instructions are not necessary. In some instances, assembly and / us use instructions can be included. In some instances, a suitable device can include a non-penetrable sheath, e.g., made of a rigid material. In some instances, the device isconfigured and made of materials such that easy and visible aspiration is possible. In some instances, the device is compact and of such a size that it can be easily handled.

[0208] In some embodiments, a device, such as but not limited to a vial or syringe, or an element to be attached to the device (e.g., a needle or tubing) can include a securing mechanism for securing one element to another (i.e., connecting the two elements). Securing mechanisms can, e.g., find use in connecting the device to a needle or to an IV delivery system, such as an IV port, a reconstitution IV bag, IV tubing or the like. Useful securing mechanisms will vary and can include any of a variety of fasteners for securing, such as but not limited to clamp mounts, clips, screws, bolts, adhesive, hook and loop tape, snap fit, friction fit, magnets, etc. In some embodiments, securing mechanisms can allow a connection to device or a needle by allowing the two elements to be threaded together via, e.g., a LUER-LOCK or other standardized threading mechanism. Delivery of the composition into the subject can be achieved by manual means (e.g., manually depressing a plunger) or mechanical means (e.g., a pump, a vacuum, etc.).

[0209] In some instances, a composition of the present disclosure can be present in a container in unit dosage form and formulated for direct or indirect delivery of a desired dose to a subject. For example, unit dosage forms for injection or intravenous administration can comprise the compound(s) in a composition as a solution in sterile water, normal saline or another pharmaceutically acceptable carrier.

[0210] One skilled in the art, based upon the disclosure provided herein, will understand that the methods and compositions of the disclosure is useful in subjects who, in whole (e.g., systemically) or in part (e.g., locally, cell, tissue, organ), are being or can be, treated for a disease or disorder where an increase in IL-18 signaling activity would be beneficial. The skilled artisan will appreciate, based upon the teachings provided herein, that the diseases and disorders treatable by the compositions and methods described herein encompass any disease or disorder wherein an increase in IL- 18 signaling will promote a positive biologic, physiologic, clinical or therapeutic outcome.KITS

[0211] The present invention also pertains to kits useful in the methods of the disclosure. Such kits comprise various combinations of components useful in any of the methods described elsewhere herein, including for example, a engineered IL-18 polypeptide or encoding nucleic acid (e.g., a stabilized IL-18 such as a stabilized DR IL-18 variant or stabilized D2D IL-18 variant, and the like), materials for IV administration of the engineered IL- 18, materials for quantitatively analyzing engineered IL- 18 polypeptide or IL- 18 variant nucleic acid, and / or materials for assessing the activity of an IL-18 variant polypeptide or an IL-18 variant nucleic acid, and / or instructional material. In various embodiments, to determine whether the level ofIL- 18 signaling in a biological sample obtained from the subject, the level is compared with the level of at least one comparator control contained in the kit, such as a positive control, a negative control, a historical control, a historical norm, or the level of another reference molecule in the biological sample.

[0212] In some embodiments, useful kits will include a composition comprising an engineered IL- 18 polypeptide or encoding nucleic acid formulated for IV delivery to a subject and one or more IV delivery devices, such as but not limited to e.g., a syringe suitable for use with a syringe pump, an IV bag, an IV cannula, an IV catheter, tubing, saline, or any combination thereof. In some instances, the composition comprising the engineered IL- 18 polypeptide or encoding nucleic acid is contained within a container or vial separate from the other components of the kit. In some instances, the composition comprising the engineered IL- 18 polypeptide or encoding nucleic acid is contained within an IV delivery component of the kit, such as e.g., an IV bag or a syringe suitable for use with a syringe pump. Such kits may or may not include one more additional components of a method described herein, such as e.g., one or more components used in CRS prophylaxis, one or more B cell depleters (e.g., obinutuzumab), or the like. Such additional component(s), when present, may be contained in any suitable container, such as a vial, syringe, etc. Elements of such kits may be configured in a read-to-administer configuration, unit dosage form, or the like.

[0213] In some embodiments, useful kits may comprise, e.g., in addition to an engineered IL-18 polypeptide or encoding nucleic acid (e.g., a stabilized IL-18 such as a stabilized DR IL-18 variant or stabilized D2D IL- 18 variant, and the like), a B-cell depleter, materials for quantitatively analyzing engineered IL- 18 polypeptide or IL- 18 variant nucleic acid or B-cell depleter, and / or materials for assessing the activity of an IL-18 variant polypeptide or an IL-18 variant nucleic acid or B-cell depleter, and / or instructional material. In various embodiments, to determine whether the level of IL-18 signaling or the level of B-cells, or ADA, in response to a B-cell depleter is modulated in a biological sample obtained from the subject, the level is compared with the level of at least one comparator control contained in the kit, such as a positive control, a negative control, a historical control, a historical norm, or the level of another reference molecule in the biological sample. Such kits may or may not further include one or more materials for IV administration of the engineered IL-18 polypeptide and / or B-cell depleter.EXAMPLES OF NON-LIMITING ASPECTS OF THE DISCLOSURE

[0214] Aspects, including embodiments, of the present subject matter described above may be beneficial alone or in combination, with one or more other aspects or embodiments. Without limiting the foregoing description, certain non-limiting aspects of the disclosure are provided below. As will be apparent to those of skill in the art upon reading this disclosure, each of theindividually numbered aspects may be used or combined with any of the preceding or following individually numbered aspects. This is intended to provide support for all such combinations of aspects and is not limited to combinations of aspects explicitly provided below.

[0215] Aspects 1-132:1. A method of delivering an engineered interleukin- 18 (IL- 18) polypeptide to a subject in need thereof, the method comprising: administering an amount of the engineered IL-18 polypeptide from 20 pg / kg to 3000 pg / kg to the subject by an intravenous (IV) route of administration, thereby delivering an effective amount of the engineered IL-18 polypeptide to the subject.2. The method of aspect 1, wherein the engineered IL- 18 polypeptide is a decoy-resistant (DR) IL- 18 polypeptide.3. The method of aspect 1 or aspect 2, wherein the engineered IL- 18 polypeptide comprises an amino acid sequence having at least 85% sequence identity with at least one of SEQ ID NOs: 1, 7, 17-22, 55-57, 62.4. The method of aspect 3, wherein the engineered IL-18 polypeptide comprises an amino acid sequence having at least 90% sequence identity with at least one of SEQ ID NOs: 1, 7, 17-22, 55-57, 62.5. The method of any one of the preceding aspects, wherein the engineered IL- 18 polypeptide comprises one or more mutations at positions Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DUO, Nl l l, Ml 13, V153, and N155 relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO: 1.6. The method of any one of the preceding aspects, wherein the engineered IL- 18 polypeptide comprises at least 5 mutations relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO: 1.7. The method of aspect 6, wherein the at least 5 mutations are at positions selected from Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DI 10, N111, Ml 13, V153, and N155.8. The method of aspect 6 or aspect 7, wherein the engineered IL-18 polypeptide comprises at least 10 mutations relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO: 1.9. The method of aspect 8, wherein the at least 10 mutations are at positions selected from Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DI 10, N111, Ml 13, V153, and N155.10. The method of any one of the preceding aspects, wherein the engineered IL- 18 polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 20, 7, 17-19, 21, 22, 55-57, 62.11. The method of any one of the preceding aspects, wherein the amount of the engineered IL- 18 polypeptide administered is from 30 pg / kg to 1200 pg / kg.12. The method of aspect 11, wherein the amount of the engineered IL-18 polypeptide administered is from 60 pg / kg to 240 pg / kg.13. The method of any of the preceding aspects, wherein the amount is administered as a single IV infusion.14. The method of any of the preceding aspects, wherein administering the effective amount of the engineered IL- 18 polypeptide comprises achieving a maximum concentration observed (Cmax) of at least 1000 ng / mL of the engineered IL- 18 polypeptide in plasma of the subject within 1 day of the administration.15. The method of aspect 14, comprising achieving a Cmax of at least 1000 ng / mL of the engineered IL-18 polypeptide in plasma of the subject within 12 hours of the administration.16. The method of aspect 15, comprising achieving a Cmax of at least 1000 ng / mL of the engineered IL-18 polypeptide in plasma of the subject within 6 hours of the administration.17. The method of any of the preceding aspects, wherein the method comprises repeating the administration weekly, once every two weeks, or once every three weeks.18. The method of aspect 17, wherein administration is repeated for at least four weeks.19. The method of aspect 18, wherein administration is repeated for at least seven weeks.20. The method of any one of aspects 17 to 19, wherein the administering results in an engineered IL- 18 polypeptide area under the curve (AUC) accumulation ratio (AR) that is at least five times higher than the engineered IL- 18 polypeptide AUC AR resulting from a corresponding subcutaneous route of administration.21. The method of any one of aspects 17 to 20, wherein the administering results in an engineered IL- 18 polypeptide Cmax AR that is at least five times higher than the engineered IL- 18 polypeptide Cmax AR resulting from a corresponding subcutaneous route of administration.22. The method of any one of the preceding aspects, wherein the subject has, following the administering, one or more reduced risks selected from the group consisting of: a reduced risk of developing an injection site reaction (ISR); a reduced risk of developing one or more symptoms of an ISR; a reduced risk of developing a Grade 3 ISR; or any combination thereof, as compared to a corresponding subject administered an equivalent amount of the engineered IL- 18 polypeptide delivered subcutaneously, intramuscularly, or intradermally.23. The method of aspect 22, wherein the equivalent amount of the engineered IL- 18 polypeptide is an equal amount of the engineered IL-18 polypeptide.24. The method of aspect 22, wherein the equivalent amount results in a physiological concentration of the engineered IL- 18 polypeptide in the subject that is analogous to the physiological concentration resulting from the amount of the engineered IL-18 polypeptide administered by the IV route of administration.25. The method of aspect 24, wherein the physiological concentration comprises a maximum concentration observed (Cmax).26. The method of any one of aspects 22 to 25, wherein the one or more symptoms of a ISR are selected from the group consisting of ulcers, necrosis, tissue damage, pain, lipodystrophy, edema, phlebitis, tenderness, erythema, itching, or any combination thereof.27. The method of any one of the preceding aspects, wherein the subject experiences fewer injection site reaction (ISR) symptoms, less severe ISR symptoms, or both, as compared to delivery of the engineered IL- 18 polypeptide subcutaneously.28. The method of aspect 27, wherein the ISR symptoms are selected from the group consisting of ulcers, necrosis, tissue damage, pain, lipodystrophy, edema, phlebitis, tenderness, erythema, itching, or any combination thereof.29. The method of any one of the preceding aspects, wherein the subject does not develop a Grade 3 ISR.30. The method of aspect 29, wherein the subject does not develop a Grade 2 or higher ISR.31. The method of aspect 30, wherein the subject does not develop an ISR.32. The method of any of the preceding aspects, wherein the subject has a cancer and the method comprises administering the engineered IL-18 polypeptide to treat the subject for the cancer.33. The method of aspect 32, wherein the cancer comprises a solid tumor.34. The method of aspect 33, wherein the cancer is selected from the group consisting of melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-high tumors, high tumor mutation burden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer.35. The method of any one of aspects 33 to 34, wherein the cancer is associated with loss of Major Histocompatibility Complex (MHC) class I surface expression.36. The method of any one of aspects 33 to 35, wherein the cancer is resistant to checkpoint inhibitor treatment.37. The method of aspect 36, wherein the cancer comprises an anti-PDl -resistant tumor.38. The method of one any of the preceding aspects, further comprising pre-treating the subject with an effective amount of a B-cell depleter prior to administering the amount of the engineeredIL- 18 polypeptide, optionally wherein the B-cell depleter comprises an anti-CD20 antibody, optionally wherein the anti-CD20 antibody is obinutuzumab.39. Use of a composition comprising an engineered IL-18 polypeptide in a method of delivering the engineered interleukin- 18 (IL- 18) polypeptide to a subject, according to any one of the preceding aspects.40. The use of aspect 39, wherein the subject has a cancer and the method comprises administering the engineered IL- 18 polypeptide to treat the subject for the cancer.41. A method of treating a subject in need thereof comprising administering an effective amount of an engineered interleukin- 18 (IL- 18) polypeptide to the subject, wherein the effective amount is administered intravenously from 20 pg / kg to 3000 pg / kg, thereby treating the subject.42. The method of aspect 41, where in the subject is a primate.43. The method of aspect 41 or 42, where in the subject is human.44. The method of any one of aspects 41 to 43, wherein the subject has a cancer.45. The method of aspect 44, wherein the cancer is a solid tumor.46. The method of aspect 44, wherein the cancer is a liquid tumor, optionally a blood cancer.47. The method of any one of aspects 44 to 46, wherein the cancer is selected from the group consisting of melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-high tumors, high tumor mutation burden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer.48. The method of any one of aspects 44 to 47, wherein the cancer is associated with loss of MHC class 1 surface expression.49. The method of any one of aspects 44 to 48, wherein the cancer comprises an anti-PD-1 resistant tumor.50. The method of one any of the preceding aspects, further comprising pre-treating the subject with an effective amount of a B-cell depleter prior to administering the amount of the engineered IL- 18 polypeptide, optionally wherein the B-cell depleter comprises an anti-CD20 antibody, optionally wherein the anti-CD20 antibody is obinutuzumab.51. Use of a composition comprising an engineered IL- 18 polypeptide, in a method of treating a subject, according to any one of aspects 44 to 50.52. The use of aspect 51, wherein the subject is a human subject having cancer and the method treats the subject for the cancer.53. A method of administering an engineered interleukin 18 (IL- 18) polypeptide to a subject in need thereof, the method comprising: pre-treating the subject with an effective amount of a B-cell depleter to deplete B-cells in the subject; and administering the engineered IL-18 polypeptide to the B-cell depleted subject.54. The method of aspect 53, wherein the engineered IL-18 polypeptide is a decoy-resistant (DR) IL-18 polypeptide.55. The method of aspect 53 or aspect 54, wherein the engineered IL-18 polypeptide comprises an amino acid sequence having at least 85% sequence identity with at least one of SEQ ID NOs:l, 7, 17-22, 55-57, and 62.56. The method of aspect 55, wherein the engineered IL-18 polypeptide comprises an amino acid sequence having at least 90% sequence identity with at least one of SEQ ID NOs: 1, 7, 17- 22, 55-57, and 62.57. The method of any one of aspects 53 to 56, wherein the engineered IL- 18 polypeptide comprises one or more mutations at positions Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DUO, Nl l l, Ml 13, V153, and N155 relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO: 1.58. The method of any one of aspects 53 to 57, wherein the engineered IL-18 polypeptide comprises at least 5 mutations relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO: 1.59. The method of aspect 58, wherein the at least 5 mutations are at positions selected from Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DI 10, N111, Ml 13, V153, and N155.60. The method of aspect 58 or aspect 59, wherein the engineered IL-18 polypeptide comprises at least 10 mutations relative to the wild-type IL- 18 amino acid sequence set forth in SEQ ID NO: 1.61. The method of aspect 60, wherein the at least 10 mutations are at positions selected from Yl, L5, K8, C38, M51, K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, DI 10, N111, Ml 13, V153, and N155.62. The method of any one of aspects 53 to 61, wherein the engineered IL- 18 polypeptide comprises an amino acid sequence selected from SEQ ID NOs:20, 7, 17-19, 22, 55-57, and 62.63. The method of any one of aspects 53 to 62, wherein the B-cell depleter is a B-cell depleting antibody.64. The method of aspect 63, wherein the antibody is an anti-CD20 antibody.65. The method of aspect 64, wherein the B-cell depleting antibody is obinutuzumab.66. The method of any one of aspects 53 to 65, wherein the subject abstains from antihypertensive drugs at least 24 hours before and at least 48 hours after administration of the engineered IL- 18 polypeptide.67. The method of any one of aspects 53 to 66, further comprising administering glucocorticoid, acetaminophen, antihistamine, or a combination thereof to the subject by an enteral or parenteral route of administration.68. The method of any one of aspects 53 to 67, wherein the pre-treating comprises administering the B-cell depleter in a single dose.69. The method of any one of aspects 53 to 67, wherein the pre-treating comprises administering the B-cell depleter in at least two separate doses.70. The method of aspect 69, wherein the B-cell depleter doses are administered at least 6 hours apart.71. The method of aspect 69 or 70, wherein the at least two separate doses are administered on consecutive days.72. The method of any one of aspects 53 to 71, wherein the pre-treating comprises administering at least 2000 mg of obinutuzumab in one or two doses.73. The method of any one of aspects 53 to 72, wherein the pretreatment is performed at least 5 days prior to administering the engineered IL-18 polypeptide.74. The method of aspect 73, wherein the pretreatment is performed at least 7 days prior to administering the engineered IL- 18 polypeptide.75. The method of any one of aspects 53 to 74, wherein the pretreatment is performed no more than 14 days prior to administering the engineered IL-18 polypeptide.76. The method of aspect 75, wherein the pretreatment is performed no more than 12 days prior to administering the engineered IL- 18 polypeptide.77. The method of aspect 76, wherein the pretreatment is performed no more than 10 days prior to administering the engineered IL- 18 polypeptide.78. The method of any one of aspects 53 to 77, wherein the pre-treatment comprises intravenous administration of the B-cell depleter.79. The method of any one of aspects 53 to 78, wherein the administering comprises intravenous administration of the engineered IL- 18 polypeptide.80. The method of any one of aspects 53 to 79, wherein the engineered IL- 18 polypeptide is administered at a dose of at least 20 pg / kg.81. The method of aspect 80, wherein the engineered IL- 18 polypeptide is administered at a dose of at least 30 pg / kg.82. The method of aspect 81, wherein the engineered IL- 18 polypeptide is administered at a dose of at least 60 pg / kg.83. The method of aspect 82, wherein the engineered IL- 18 polypeptide is administered at a dose of at least 120 pg / kg.84. The method of aspect 83, wherein the engineered IL-18 polypeptide is administered at a dose of at least 240 pg / kg.85. The method of any one of aspects 53 to 84, wherein the subject has not been previously administered the engineered IL-18 polypeptide.86. The method of any of aspects 53 to 84, wherein the subject has been previously administered the engineered IL- 18 polypeptide.87. The method of aspect 86, wherein the subject has detectable levels of anti-drug antibodies to the engineered IL- 18 polypeptide.88. The method of aspect 87, further comprising testing for the presence of anti -drug antibodies to the engineered IL- 18 polypeptide in a biological sample obtained from the subject.89. The method of any one of aspects 53 to 88, wherein the subject has a cancer and the method comprises administering the engineered IL-18 polypeptide to treat the subject for the cancer.90. The method of aspect 89, wherein the cancer is not a blood cancer.91. The method of aspect 90, wherein the cancer is not a B-cell neoplasm.92. The method of aspect 91, wherein the cancer is not a lymphoma.93. The method of aspect 92, wherein the cancer is not non -Hodgkin's lymphoma.94. The method of any one of aspects 90 to 93, wherein the cancer is a solid tumor.95. The method of aspect 94, wherein the solid tumor does not express CD20.96. The method of aspect 89 or 95, wherein the cancer is selected from the group consisting of melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-high tumors, high tumor mutation burden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer.97. The method of any one of aspects 89 to 96, wherein the cancer is associated with loss of surface expression of Major Histocompatibility Complex (MHC) class I.98. The method of any one of aspects 89 to 97, wherein the cancer is resistant to checkpoint inhibitor treatment.99. The method of aspect 98, wherein the cancer comprises an anti-PDl resistant tumor.100. Use of a composition comprising an engineered IL- 18 polypeptide, in a method of administering the engineered IL- 18 polypeptide to a subject, according to any one of aspects 53 to 99.101. The use of aspect 100, wherein the subject is a human subject having cancer and the method treats the subject for the cancer.102. A method of treating a subject in need thereof, the method comprising: pre-treating the subject with an effective amount of a B-cell depleter to deplete B-cells in the subject; and administering an engineered interleukin (IL- 18) polypeptide to the B-cell depleted subject, thereby treating the subject.103. The method of aspect 102, where in the subject is human.104. The method of aspect 102 or 103, wherein the subject has a cancer.105. The method of aspect 104, wherein the cancer is a solid tumor.106. The method of aspect 104 or 105, wherein the cancer is selected from the group consisting of melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-high tumors, high tumor mutation burden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer.107. The method of any one of aspects 104 to 106, wherein the cancer is associated with loss of Major Histocompatibility Complex (MHC) class 1 surface expression.108. The method of any one of aspects 104 to 107, wherein the cancer comprises an anti-PD-1 resistant tumor.109. Use of a composition comprising an engineered IL- 18 polypeptide, in a method of treating a subject, according to any one of aspects 102 to 108.110. The use of aspect 109, wherein the subject is a human subject having cancer and the method treats the subject for the cancer.111. The method or use of any one of the preceding aspects, further comprising administering CRS prophylaxis to the subject prior to, or while, administering the engineered IL- 18 polypeptide.112. The method or use of aspect 111, wherein the CRS prophylaxis comprises at least a dose of a NSAID and at least a dose of a histamine Hl antagonist; at least a dose of acetaminophen and at least a dose of a histamine Hl antagonist; at least a dose of a NSAID and at least a dose of acetaminophen; or at least a dose of a NSAID, at least a dose of a histamine Hl antagonist, and at least a dose of acetaminophen; optionally wherein the NSAID is selected from indomethacin and ibuprofen and the histamine Hl antagonist is diphenhydramine.113. The method or use of aspect 112, wherein:(i) the NSAID is oral indomethacin administered at 50 mg to 100 mg per day, optionally at 75 mg per day, or oral ibuprofen administered at 600 mg to 2,400 mg per day, optionally 800 mg to 1,800 mg per day;(ii) the histamine Hl antagonist is intravenous or oral diphenhydramine administered at a dose of 25 mg to 100 mg, optionally 50 mg(iii) the acetaminophen is oral acetaminophen administered at 350 mg to 4,000 mg per day, optionally 650 mg to 3,900 mg per day; or(iv) any combination thereof.114. A method of delivering an engineered interleukin- 18 (IL-18) polypeptide to a subject in need thereof, the method comprising: administering an amount of the engineered IL-18 polypeptide from 20 pg / kg to 3000 pg / kg to the subject by an intravenous (IV) route of administration, thereby delivering an effective amount of the engineered IL-18 polypeptide to the subject.115. The method of aspect 114, wherein the engineered IL- 18 polypeptide is a decoy-resistant (DR) IL- 18 polypeptide, optionally wherein the DR IL- 18 polypeptide has less than 100% sequence identity and at least 85% sequence identity to WT human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO: 102), wherein the X at position (“pos.”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H.116. The method of aspect 115, wherein the DR IL- 18 polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 20, 7, 17-19, 21, 22, 55-57, and 62, optionally wherein the DR IL-18 polypeptide consists of, or consists essentially of, an amino acid sequence selected from SEQ ID NOs: 20, 7, 17-19, 21, 22, 55-57, and 62.117. The method of any one of aspects 114 to 116, wherein the amount of the engineered IL- 18 polypeptide administered is from 30 pg / kg to 1200 pg / kg, optionally from 60 pg / kg to 240 Pg / kg-118. The method of any of aspects 114 to 117, wherein the amount is administered as a single IV infusion.119. The method of any of aspects 114 to 118, wherein administering the effective amount of the engineered IL- 18 polypeptide comprises achieving a maximum concentration observed (Cmax) of at least 1000 ng / mL of the engineered IL- 18 polypeptide in plasma of the subject within 1 day of the administration, optionally within 12 hours of the administration, optionally within 6 hours of the administration.120. The method of any of aspects 114 to 119, wherein the method comprises repeating the administration weekly, once every two weeks, or once every three weeks, optionally wherein administration is repeated for at least four weeks or at least seven weeks.121. The method of any one of aspects 114 to 120, wherein the administering results in an engineered IL- 18 polypeptide area under the curve (AUC) accumulation ratio (AR) that is at least five times higher than the engineered IL- 18 polypeptide AUC AR resulting from a corresponding subcutaneous route of administration, an engineered IL- 18 polypeptide Cmax AR that is at least five times higher than the engineered IL- 18 polypeptide Cmax AR resulting from a corresponding subcutaneous route of administration, or both.122. The method of any one of aspects 114 to 121, wherein the subject has, following the administering, one or more reduced risks selected from the group consisting of: a reduced risk of developing an injection site reaction (ISR); a reduced risk of developing one or more symptoms of an ISR; a reduced risk of developing a Grade 3 ISR; or any combination thereof, as compared to a corresponding subject administered an equivalent amount of the engineered IL- 18 polypeptide delivered subcutaneously, intramuscularly, or intradermally.123. The method of any one of aspects 114 to 122, wherein the subject experiences fewer injection site reaction (ISR) symptoms, less severe ISR symptoms, or both, as compared to delivery of the engineered IL- 18 polypeptide subcutaneously, optionally wherein the ISR symptoms are selected from the group consisting of ulcers, necrosis, tissue damage, pain, lipodystrophy, edema, phlebitis, tenderness, erythema, itching, or any combination thereof.124. The method of any one of aspects 114 to 123, wherein the subject does not develop a Grade 3 ISR, does not develop a Grade 2 or higher ISR, or does not develop an ISR.125. The method of any of aspects 114 to 124, wherein the subject has a cancer and the method comprises administering the engineered IL-18 polypeptide to treat the subject for the cancer, optionally wherein the cancer comprises a solid tumor, optionally wherein the cancer is selected from the group consisting of melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-high tumors, high tumor mutation burden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer, optionally wherein the cancer is associated with loss of Major Histocompatibility Complex (MHC) class I surface expression, optionally wherein the cancer is resistant to checkpoint inhibitor treatment, optionally wherein the cancer comprises an anti-PDl -resistant tumor.126. The method of one any of aspects 114 to 125, further comprising pre-treating the subject with an effective amount of a B-cell depleter prior to administering the amount of the engineered IL- 18 polypeptide, optionally wherein the B-cell depleter comprises an anti-CD20 antibody, optionally wherein the anti-CD20 antibody is obinutuzumab.127. A method of administering an engineered interleukin 18 (IL-18) polypeptide to a subject in need thereof, the method comprising:pre-treating the subject with an effective amount of a B-cell depleter to deplete B-cells in the subject; and administering the engineered IL-18 polypeptide to the B-cell depleted subject, optionally wherein the engineered IL-18 polypeptide is a decoy-resistant (DR) IL-18 polypeptide, optionally wherein the B-cell depleter is a B-cell depleting antibody, optionally an anti-CD20 antibody, optionally obinutuzumab.128. The method of aspect 127, wherein the administering comprises intravenous administration of the engineered IL- 18 polypeptide, optionally wherein the engineered IL- 18 polypeptide is administered at a dose of at least 20 pg / kg, at least 30 pg / kg, at least 60 pg / kg, at least 120 pg / kg, or at least 240 pg / kg.129. The method of aspect 127 or 128, wherein the subject has a cancer and the method comprises administering the engineered IL-18 polypeptide to treat the subject for the cancer, optionally wherein the cancer is a solid tumor.130. The method of any one of aspects 114 to 129, further comprising administering CRS prophylaxis to the subject prior to, or while, administering the engineered IL- 18 polypeptide.131. Use of a composition comprising an engineered IL- 18 polypeptide, in a method of treating a subject, according to any one of aspects 114 to 130, wherein the subject is a human subject having cancer and the method treats the subject for the cancer.132. A kit comprising: a pharmaceutical composition formulated for intravenous (IV) delivery to a subject, the pharmaceutical composition comprising an engineered IL- 18 polypeptide and a pharmaceutically acceptable carrier; and one or more IV delivery devices, optionally wherein the one or more IV delivery devices are selected from the group consisting of a syringe suitable for use with a syringe pump, an IV bag, an IV cannula, an IV catheter, and any combination thereof.SEQUENCESEXAMPLES

[0216] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for.Example 1 - Pharmacokinetic behaviors of Test Compound in non-human primates (NHPs)

[0217] The human cytokine interleukin 18 (IL- 18) has been previously investigated for the treatment of cancer in human clinical trials as recombinant human IL-18 (rhIL-18). However, clinical development of rhIL-18 has been curtailed by its lack of efficacy due, at least in part, to negative regulation by a decoy receptor called IL- 18 binding protein (IL-18BP). Accordingly,engineered versions of IL-18 that maintain IL-18 signaling but are resistant to inhibition by IL- 18BP are under development. In humans, rhIL-18 was found to be safe and well-tolerated.

[0218] The Test Compound is an engineered IL- 18 cytokine that is not inhibited by IL-18BP and retains activity in human IL-18R reporter assays. The Test Compound has been specifically engineered to minimize inhibition by the decoy IL-18 receptor, IL-18BP, that is highly prevalent in the tumor microenvironment. For this reason, the Test Compound is anticipated to demonstrate considerably improved efficacy compared to wild-type recombinant IL- 18. Preclinical data in syngeneic mouse models both as monotherapy and in combination with checkpoint inhibitors show activity in immunogenic (“hot”) and checkpoint inhibitor resistant (“cold”) tumors. Since most patients treated with checkpoint inhibitors fail to derive significant clinical benefit from current immunotherapy, this molecule has the potential to fulfill a high unmet medical need in solid tumors.Objectives

[0219] The objectives of the study included evaluating the pharmacokinetic / toxicokinetic characteristics and potential toxicity of the Test Compound when administered by subcutaneous (SC) and intravenous (IV) injection once weekly on Days 1, 8, 15, and 22 to non-human primates (NHPs), and to evaluate the potential reversibility of any findings following a 2-week recovery period.Materials and Methods

[0220] The Test Compound was used at 30 mg / mL to prepare dose formulations and formulations were filtered using a 0.2 pm polyvinylidene fluoride (PVDF) filter prior to dose administration. Purpose-bred naive cynomolgus monkeys (Macaca fascicularis) of 2 to 4 years of age and weighing 1.8 to 4.0 kg at initiation dosing were employed. The study experimental design was as presented in the following table:

[0221] The dose formulations were administered to appropriate animals in Group 1 by subcutaneous injection into the interscapular area once weekly on Days 1, 8, 15, and 22. While the lowest dose employed was 0.1 mg / kg, it is notable that the Test Compound was seen to be active in preclinical studies at much lower doses, such as e.g., 10 pg / kg. On each dosing day, the SC administration in Group 1 animals was followed by an intravenous infusion over 60 minutes (±6 minutes) starting within 5 minutes of completion of the SC dose. Dose formulation was administered using a temporary catheter inserted into a peripheral vein connected to a primed infusion line. The appropriate volume was delivered using an infusion pump.

[0222] The dose formulations were administered to appropriate animals in Groups 2 to 4 by subcutaneous injection into the interscapular area once weekly on Days 1, 8, 15, and 22.

[0223] The dose formulations were administered to appropriate animals in Group 5 by intravenous infusion over 60 minutes (±6 minutes) once weekly on Days 1, 8, 15, and 22. Dose formulation was administered using a temporary catheter inserted into a peripheral vein connected to a primed infusion line. The appropriate volume was delivered using an infusion pump.

[0224] Dose administration was completed within 6 hours from the removal of formulations from the refrigerator and within 90 hours from the formulation preparation completion time. Various experimental observations and measurements were performed, including mortality / morbidity checks, food evaluations, cage side observations, detailed examinations, body weight measurements, ophthalmology, body temperature measurements, and el ectrocardiography .

[0225] For clinical pathology, animals were fasted for at least four hours before sample collections. Blood samples were collected from an appropriate peripheral vein other than the one used for dosing for up to 24 hours post dosing. Blood samples were collected once from all animals during acclimation and at days 2, 29, and 47 for all animals in groups 1 to 5. Numerous hematology and serum chemistry parameters were assessed.

[0226] For laboratory analyses (e.g., toxicokinetic (TK) evaluation, anti-drug antibody (ADA) analysis, cytokine analysis, iron panel analysis, etc.) blood was collected from all animals in groups 1 to 5 on Day 1 at pre-dose and 0.25, 0.5, 1, 1.5, 4, 6, 9, 12, 24, and 48 hours post-dose; on Day 15 pre-dose; and on Day 22 pre-dose and 0.25, 0.5, 1, 1.5, 4, 6, 9, 12, 24, and 48 hours post-dose. Post-dose for Groups 2 to 4 was following the end of injection and post-dose for Groups 1 and 5 was following the start of infusion. TK sample processing followed standardprocedures, including refrigerated centrifugation within 60 minutes of collection, aliquoting, storage, and shipment on dry ice, and testing (and retesting where necessary) by a qualified laboratory. TK parameters were computed for Day 1 and 22.Results

[0227] Blood samples were collected and plasma concentrations of the Test Compound were determined as described. FIG. 1 provides the plasma concentrations of the Test Compound in each individual animal over the first six hours of Day 1 following administration of a 1 mg / kg dose of the Test Compound by either subcutaneous injection (SC) or intravenous infusion (IV). As is readily apparent from FIG. 1, plasma concentrations of the Test Compound were observed to be substantially higher following IV administration as compared to SC administration.

[0228] TK parameters after single and repeat SC or IV administration of the Test Compound every week for 4 weeks were obtained by noncompartmental analysis (NCA) of the plasma concentration data sets. A summary of the maximum concentration observed (Cmax) parameters, representing the maximum observed concentration measured after dosing, for male and female animals following a single (Day 1) or repeat (Day 22) 1 mg / kg dosing by SC or IV route of administration (ROA) (i.e., Groups 4 and 5) is provided in the following table:

[0229] A summary of the AUC parameters for male and female animals following single (Day1) and repeat (Day 22) 1 mg / kg dosing by SC or IV ROA (i.e., Groups 4 and 5) is provided in the following table:

[0230] The provided AUC values, include AUCiast : representing the area under the concentration-versus-time curve from the start of dose administration to the time after dosing atwhich the last quantifiable concentration was observed, using the linear trapezoidal method; AUCinf : representing the area under the plasma concentration time curve extrapolated to infinity, calculated as AUCiast + Ciast / z , where C is the measured concentration at time Tiast (i.e., the time after dosing at which the last quantifiable concentration was observed); and AUCT: representing the area under the concentration versus time curve calculated over the dosing interval (T = 168 hours), using the linear trapezoidal method. AUCiast and AUCinf are calculated at Day 1 dosing only and AUCTis calculated at the Day 22 dose. No marked sex differences (<2 fold) between males and females were observed for Cmax and AUCiast following a single SC or IV administration of the Test Compound.

[0231] Following repeated dosing, the AUCTvalues for IV admnistration were substantially higher than SC administration. These data represent an overall actual exposure to the Test Compound by IV administration that was higher than SC administration and to a degree that was unexpected. This surprising difference was also reflected in higher AUC Accumulation Ratios (AR) (AUCiast, Day22 / AUCiast, Dayi) for IV administration (1.7 (male), 3.5 (female)) versus SC administration (0.15 (male), 0.033 (female)) as well as the higher Cmax AR (Cmax, Day22 / Cmax, Dayi) for IV (1.4 (m), 1.8 (f)) versus SC (0.15 (m), 0.067 (f)).

[0232] A profound treatment-related anti-drug antibody (ADA) reaction to the Test Compound has been observed in NHPs. All animals treated with the Test Compound tested positive to AD As on Day 15 and / or Day 22 collections. Without being bound by theory, the presence of AD As likely impacts the measured PK / TK parameters of the Test Compound. For example, the presence of AD As appeared to impact TK parameters estimation in all animals administered the Test Compound subcutaneously, as evidenced by a steeper decline of the terminal elimination phase and increased clearance on Day 22.

[0233] Collectively, these data display differences in the pharmokinetic behaviours of the Test Compound administered by IV ROA and SC ROA. These data further highlight opportunities to achieve substantially increased maximum concentrations within patient plasma through IV administration while also increasing patient exposure to the Test Compound through the course of treatment. Moreover, higher dose levels (3.2 and 10 mg / kg) of a different, but structurally related, test compound were not tolerated in monkeys repeat-dosed, indicating that the difference in PK parameters observed between SC and IV dosing may not be overcome merely by, e.g., increasing the dose of an engineered IL-18 polypeptide administered by the SC ROA until parameters more similar to those obtained by IV dosing are achieved. The degree to which the Test Compound performed differently when administered to NHPs by these different RO As was unexpected and could not have been predicted a priori.Example 2 - Injection-site reactions following subcutaneous administration of the Test Compound in NHPs

[0234] Analysis of NHPs treated with the Test Compound by SC ROA, identified histopathological changes in a few animals at the terminal necropsy. In particular, mild fibroplasia (1 male at 0.3 mg / kg and 1 female at 1 mg / kg) and / or mild mononuclear cell infiltration (1 male at 0.1 mg / kg, 1 female each at 0.3 and 1.0 mg / kg) were observed at the SC injection site, compared to controls. No animals had mild or greater mononuclear cell infiltration at the injection sites in recovery animals, and no recovery animals had fibroplasia at the injection sites. These findings were not considered adverse and no gross pathology findings were observed at the injection site(s) for any animals that received the Test Compound and no adverse microscopic findings were observed at the SC injection sites.

[0235] Preclinical toxicokinetic studies indicated that the Test Compound can be safely administered to non-human primates through both the SC and IV routes at doses of up to 1 mg / kg. Both routes of administration produced similar single dose AUC of drug exposure, but exhibited differing PK profiles.Example 3 - Injection-site reactions following subcutaneous administration of the Test Compound in adult humans

[0236] A study administering the Test Compound by SC ROA was initiated in adult subjects with solid tumors who have developed disease progression through standard therapy, or for whom standard of care therapy that prolongs survival is unavailable or unsuitable.

[0237] The Test Compound was supplied as a sterile solution for injection. Subjects were scheduled to receive weekly treatment with the Test Compound, with a less intensive schedule permitted based on a subject’s tolerance of toxi cities. Subjects were to be treated until one or more various endpoints were reached, including but not limited to, e.g., disease progression, achievement of maximal response, etc., or a decision was made to discontinue treatment. The starting dose of the Test Compound was 30 pg / kg.

[0238] During investigational treatment, multiple subjects enrolled who received the Test Compound by SC ROA experienced a reaction at the site of injection with or without itching. One study subject experienced a moderate-severe reaction when the Test Compound was given, which resulted in an ulcer at the injection site. The occurrence of injection-site reactions was unexpected, at least in part due to the lack of gross or microscopic injection-site reactions in NHPs administered the Test Compound by SC ROA.

[0239] The prevalence of injection site reaction in response to biologies varies widely, with one meta-analysis review of 158 articles reporting prevalences ranging from 0.08% to 15.5% for 16 different biologies across 80 clinical studies (Kim PJ, et al. A Systematic Review and Meta-Analysis of Injection Site Reactions in Randomized-Controlled Trials of Biologic Injections. Journal of Cutaneous Medicine and Surgery. 2023;27(4):358-367). ISRs may have an impact on patient satisfaction with treatment and contribute to reasons for treatment discontinuation.Efforts to address ISR include reformulation of biologies with different buffers, such as e.g., a citrate-free formulation of Ixekizumab (Idf.

[0240] Over the course of the SC monotherapy trial, the Test Compound was found to be generally well tolerated. Injection site reactions were the most frequently observed treatment- related adverse events (AEs), occurring in 55% of subjects overall and in all dose cohorts (30 pg / kg, 60 pg / kg, 120 pg / kg, and 240 pg / kg). Injection site reactions were graded according to CTCAEv5:Grade 1 - Tenderness with or without associated symptoms (e.g., warmth, erythema, itching) Grade 2 - Pain; lipodystrophy; edema; phlebitisGrade 3 - Ulceration or necrosis; severe tissue damage; operative intervention indicated.

[0241] In the highest two dose cohorts (120 pg / kg, and 240 pg / kg), 10% of those subjects that experienced an ISR AE experienced a Grade 3 reaction. All patients in these higher dose cohorts, as well as 93% of all subjects in the SC trial, received CRS prophylaxis. Accordingly, while CRS prophylaxis was not intended to prevent the occurrence of ISRs, the above-described level of ISRs was nevertheless observed regardless of the pre-administration of CRS prophylaxis medications, which can include steroids, NSAIDs, acetaminophen, and histamine Hl and / or H2 antagonists / blockers, to the subjects shortly before SC injection of the Test Compound.

[0242] Collectively, these findings emphasize the unexpected nature of ISRs observed following SC administration of the engineered IL- 18 polypeptide Test Compound.Example 4 - Intravenous administration of the Test Compound in human clinical studies

[0243] Delivery of the Test Compound by an IV route of administration is expected to provide not only an advantageous PK profile (with IV administration exhibiting higher Cmax values for the same AUC), but also is expected to benefit patient comfort and safety through the reduction of ISRs compared to delivery by SC route of administration.Objectives

[0244] Objectives of the study include but are not limited to: determining the maximum tolerated dose (MTD) and recommended future study dosing of Test Compound in test subjects with solid tumors (including relapsed and refractory solid tumors); evaluating the anti-tumor activity of the Test Compound; evaluating the overall safety and tolerability of the Test Compound; and characterizing the pharmacokinetic (PK) and pharmacodynamic (PD) profile of the Test Compound.Materials and Protocols

[0245] This study is a dose escalation monotherapy study of the Test Compound. Some exact parameters for later portions of the planned study, e.g., with respect to formulation and pretreatment, may be determined after completion of study elements in earlier phases.

[0246] The study is performed in adult test subjects with solid tumors who have developed disease progression through standard therapy, or for whom standard of care therapy that prolongs survival is unavailable or unsuitable. All test subjects are pre-medicated for cytokine release syndrome (CRS) prior to dosing with the Test Compound.

[0247] The Test Compound is supplied as a sterile solution for injection to be administered by SC or IV injection. Test subjects initially receive weekly treatment with the Test Compound. A less intensive schedule is permissible based on a test subject’s tolerance of toxi cities. Additional dosing schedules are tested based on emerging safety, tolerability, PK, and preliminary antitumor activity of the Test Compound. Test subjects are treated until one or more various endpoints are reached, including but not limited to, e.g., disease progression, achievement of maximal response, etc., or a decision is made to discontinue treatment.

[0248] The starting dose of the Test Compound is 30 pg / kg or 60 pg / kg, with a total of 6-7 dose level cohorts that will escalate in the absence of >Grade 2 treatment-related AEs. Upon reviewing data from all test subjects, evaluation of the Test Compound higher doses, including, e.g., doses beyond 1200 pg / kg in increments of 33%, are considered.

[0249] The starting dose for the IV monotherapy dosing is modeled to have a maximum concentration observed (Cmax) that does not exceed the Cmax for the highest tolerated SC dose approved by the Independent Safety Review Committee (ISRC). The 60 pg / kg IV starting dose is expected to reproduce the Cmax of 120 pg / kg administered SC. The Test Compound is given on a weekly, biweekly (every two weeks), triweekly (every three weeks), or monthly schedule when administered by the IV route of administration. Escalation decisions for the IV formulation will be based primarily on its emerging toxicity profile, although information from the SC dosing will also be taken into account. Dose magnitude, dosing increments and the dosing schedule can be modified as additional safety, tolerability, PK, and preliminary antitumor activity data for the Test Compound become available.

[0250] Pre-treatment medication, including for CRS prophylaxis, can include: administering an oral nonsteroidal anti-inflammatory drug (NS AID) 1-2 hours prior to therapy and continuing for at least 48 hours post-dose. Choice of NS AID can include but not be limited to indomethacin 25 mg 3 times daily or oral ibuprofen 200 to 600 mg every 6 to 8 hours; adding oral acetaminophen 650 mg every 4 to 6 hours as needed for the first 24 hours post-dose; for test subjects with contraindication to NSAIDs, oral acetaminophen is administered 650 mg every 4 to 6 hoursbeginning 1 hour prior to dosing with the Test Compound and continued for at least 48 hours post-dose; administering IV or oral diphenhydramine 50 mg or a comparable dose of a nextgeneration histamine Hl antagonist 30-60 minutes prior to dosing with the Test Compound; for test subjects with or at risk for hypotension, administering of up to 3 liters of oral / IV fluids in the 24 hours post-dose; where indicated for subsequent doses, administering famotidine (or other H2 blocker) at 20 mg IV or 20-40 mg orally, 30-60 minutes prior to dosing with the Test Compound.

[0251] Prophylactic antibiotics are administered as medically necessary, and test subjects will receive therapeutic antibiotics for intercurrent infections as needed.

[0252] The Test Compound is administered weekly, biweekly, triweekly, or monthly, with dosing schedules adjusted based on emerging safety, tolerability, PK, and preliminary antitumor activity of the Test Compound. The Test Compound is administered as a sterile solution for injection by SC injection or IV infusion.

[0253] Concomitant medications are administered where deemed medically necessary, including e.g., transfusion of red blood cells (RBCs) or platelets and administration of erythropoietin or thrombopoietin; myeloid growth factors (e.g., granulocyte-colony-stimulating factor [G-CSF]) for test subjects with febrile neutropenia (temperature >38.5°C), non-febrile neutropenia with infection, or an ANC <500 / mm3. Routine prophylaxis with myeloid growth factors is avoided unless there is a documented history of febrile neutropenia or non-febrile neutropenia with infection. Growth factor administration is withheld unless a significant risk of safety in the face of peripheral neutropenia is present.

[0254] Other concomitant medications will be administered where deemed medically necessary, including: prophylactic treatment with anti -infective drugs (e.g., antibiotics, antifungals) and nonsteroidal anti-inflammatory drugs (NSAIDs) or corticosteroids (up to 10 mg QD) are permitted.

[0255] Anti-emetic and anti-diarrheal prophylaxis is withheld unless a test subject has experienced >Grade 1 CTCAE symptoms.

[0256] Test subjects will receive hematopoietic support where indicated, including: granulocyte colony-stimulating factor (G-CSF; e.g., filgrastim, filgrastim-sndz, pegfilgrastim, lenograstim) administered in response to Grade >3 neutropenia or neutropenic complications after Cycle 1; erythropoietic agents (e.g., erythropoietin or darbepoetin) administered for Grade >3 anemia; and red blood cell or platelet transfusions are administered as medically indicated.Test Subjects and Expected Results

[0257] Test subjects will be treated with the Test Compound as described above. Test subjects will include those having solid tumors, including those having a histologically or cytologicallyconfirmed diagnosis of advanced / metastatic solid tumor, including test subjects having solid tumors such as Melanoma, Merkel cell, renal cell carcinoma (RCC), urothelial, non-small lung cancer (NSCLC; inc., with no epidermal growth factor receptor (EGFR), tropomyosin receptor kinase (TRK) receptor, or anaplastic lymphoma kinase (ALK) positive mutations / fusions), triple-negative breast cancer (TNBC), squamous cell carcinoma of head and neck (SCCHN), microsatellite instability high (MSI-Hi) tumors, high tumor mutation burden (Hi-TMB) or mismatch repair deficient, gastric, cervical, endometrial, cutaneous squamous, small cell lung, esophageal, hepatocellular carcinoma (HCC) and platinum resistant ovarian cancer. In some instances, test subjects can include checkpoint-naive test subjects, including e.g., those having platinum-resistant ovarian cancer or microsatellite-stable colorectal cancer.

[0258] Various assessments, including of safety and efficacy, will be regularly performed during the treatment. Test subjects will be monitored for the incidence of dose-limiting toxi cities (DLTs) and AEs, changes in clinical laboratory parameters, vital signs, electrocardiogram (ECG) parameters, physical examination findings, and the incidence of anti -drug antibodies (ADA) and neutralizing antibodies. Accordingly, blood and plasma samples will be collected and utilized in test subject monitoring, and also PK and PD, as well as other analyses.

[0259] Disease assessments corresponding to accepted disease assessment criteria for the test subject’s particular disease (e.g., type of cancer / tumor) will be regularly performed, including on monthly, 6-week, 8-week, or 12-week intervals. Disease assessments can include imaging, such as CT and / or MRI imaging.

[0260] Tumor biopsies will be collected at various timepoints, including e.g., at screening, at ± 3 days prior to the start of Cycle 2, coincident with the first tumor assessment, well before the first tumor assessment during routine on-treatment biopsy, and the like. For test subjects that have disease progression, it is expected that one or more optional additional biopsies can be obtained at the time of disease progression. For test subjects having an accessible cutaneous lesion at baseline, a complete resection of the lesion will be performed at End of Treatment (EOT).

[0261] Various assessments of test subject blood samples will also be performed (e.g., measuring test subject PK parameters, measuring for the presence and / or titer of ADA present in test subject plasma, and the like). Blood samples for Test Compound PK and ADA analysis will be collected from all test subjects as described. Serial PK samples will be collected at multiple scheduled sampling times. Plasma concentrations of the Test Compound will be measured for each PK sample using validated method(s). Pharmacokinetic parameters such as maximum concentration observed (Cmax ), time at which maximal concentration was observed (tmax), AUCo-t (area under the concentration-time curve from time zero to time t), area under the concentration-time curve during the dosing interval (AUCT), AUCo- / (area under theconcentration-time curve from time zero extrapolated to infinity), and ti / 2 (elimination half-life) will be estimated from plasma concentration-time data. Samples will be collected for PK assessments at: C1D1 pre-dose (0-24 hrs), C1D1 post-dose 1 hr (±10 min.), C1D1 post-dose 2 hrs (±10 min.), C1D1 post-dose 2.5 hrs (±10 min.), C1D1 post-dose 3 hrs (±10 min.), C1D1 post-dose 4 hrs (±30 min.), C1D1 post-dose 6 hrs (±30 min.), C1D1 post-dose 8 hrs (±30 min.), C1D8 pre-dose 0-24 hrs, C1D8 (post-dose 4 hrs ±30 min.), C1D15 pre-dose 0-24 hrs, C1D15 (post-dose 4 hrs ±30 min.), C1D22 pre-dose 0-24 hrs, C1D22 (post-dose 4 hrs ±30 min.), C2D1 (post-dose 4 hrs ±30 min.), C2D15 (post-dose 4 hrs ±30 min.), and Day 1 of any subsequent cycles (post-dose 4 hrs ±30 min.). Based on emerging PK data, the actual collection times can change.

[0262] Efficacy responses will be assessed throughout treatment, which will include objective response based on test subjects having a CR or PR by RECIST Version 1.1. Also assessed will be best objective response rate (ORR), disease control rate (DCR) (CR, PR, or stable disease (SD) for >12 weeks), duration of response (DOR), time to response (TTR), PFS, and OS, as measured from the date of first study drug dose to the date of death by any cause.

[0263] Test subjects are treated until disease progression, death, unacceptable toxicity, symptomatic deterioration, achievement of maximal response, the Investigator’s decision to discontinue treatment, test subject decision to discontinue treatment or withdraw consent, lost to follow-up, or the sponsor decides to terminate the study.

[0264] It is expected that test subjects treated with the Test Compound by IV ROA will demonstrate less (and / or less severe) ISRs, e.g., as compared to subjects treated with the Test Compound by SC ROA (either in a concurrent study arm or from reference data). For example, it is expected that IV ROA will result in: less injection site ulcers, fewer Grade 3 ISRs, as well as increased patient comfort and safety as compared to subjects administered the Test Compound by SC ROA. It is expected that test subjects administered the Test Compound by IV ROA will experience not only improved safety and comfort through a reduction in ISRs, but also advantageous PK profiles as compared to SC ROA.

[0265] When treated with the Test Compound by IV ROA, it is expected that test subjects will demonstrate improved measures of anti-tumor activity, improved treatment biomarkers, and / or improved efficacy, e.g., by one or more of the assessments described herein. Such improvements can be relative to a concurrent study arm not employing IV ROA, reference data where IV ROA was not employed, and / or what would have been expected from the pharmacokinetics observed during Test Compound administration by SC ROA.

[0266] Disease assessments will be made using a CT or MRI scan of the chest, abdomen, and pelvis (and other relevant areas if they include target lesions). Determinations of the magnitudeand duration of changes in tumor size will be based on well-established response and progression criteria (Response Evaluation Criteria in Solid Tumors (RECIST) Version 1.1; see Eisenhauer et al. (2009) European Journal of Cancer 45:228-247) as applied to radiographic measurements. Disease assessments will be done every 6 weeks for the first 24 weeks then every 12 weeks. Clinical laboratory, pharmacodynamic, pharmacogenomic, pharmacokinetic, and other assessments will also be performed.

[0267] The cohort is expected to display evidence of pharmacodynamic activity as well as substantial evidence of tumor regressions including objective responses (CR and PR) as assessed by RECIST Version 1.1. Stable disease for > 12 weeks will be considered clinically relevant in this population of patients.Example 5 - Initial IV ROA dosing of subjects with Test Compound

[0268] A human trial cohort, for evaluating IV administration of the Test Compound, has been initiated. Patients have been administered ST-067 by IV administration through dilution of 30 mg / mL Test Compound into a 50 mL saline bag to provide an initial starting dose of 60 pg / kg. At data cutoff, two patients had received a single dose (C1D1) and one patient had received three doses (C1D15). No Grade 3 ISRs have been reported following administration of the Test Compound to subjects in this IV administration cohort.

[0269] In at least some of the previously described embodiments, one or more elements used in an embodiment can interchangeably be used in another embodiment unless such a replacement is not technically feasible. It will be appreciated by those skilled in the art that various other omissions, additions and modifications can be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modifications and changes are intended to fall within the scope of the subject matter, as defined by the appended claims.

[0270] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as “open” terms (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases “at least one” and “one or more” to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles “a” or “an” limits any particular claim containing suchintroduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases “one or more” or “at least one” and indefinite articles such as “a” or “an” (e.g., “a” and / or “an” should be interpreted to mean “at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of “two recitations,” without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to “at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “ a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and / or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase “A or B” will be understood to include the possibilities of “A” or “B” or “A and B.”

[0271] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0272] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1, 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1, 2, 3, 4, or 5 articles, and so forth.

[0273] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes andmodifications can be made thereto without departing from the spirit or scope of the appended claims.

[0274] Accordingly, the preceding merely illustrates the principles of the invention. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art, and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the claims.

[0275] The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims. In the claims, 35 U.S.C. §112(f) or 35 U.S.C. §112(6) is expressly defined as being invoked for a limitation in the claim only when the exact phrase "means for" or the exact phrase "step for" is recited at the beginning of such limitation in the claim; if such exact phrase is not used in a limitation in the claim, then 35 U.S.C. § 112 (f) or 35 U.S.C. §112(6) is not invoked.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A method of delivering an engineered interleukin- 18 (IL- 18) polypeptide to a subject in need thereof, the method comprising: administering an amount of the engineered IL-18 polypeptide from 1 pg / kg to 3000 pg / kg to the subject by an intravenous (IV) route of administration, thereby delivering an effective amount of the engineered IL- 18 polypeptide to the subject.

2. The method of claim, wherein the amount of the engineered IL- 18 polypeptide is from 20 pg / kg to 1200 pg / kg.

3. The method of claim 1 or claim 2, wherein the engineered IL-18 polypeptide is a decoyresistant (DR) IL- 18 polypeptide, optionally wherein the DR IL- 18 polypeptide has less than 100% sequence identity and at least 85% sequence identity to WT human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence:XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO: 102), wherein the X at position (“pos.”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H.

4. The method of claim 3, wherein the DR IL-18 polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 20, 7, 17-19, 21, 22, 55-57, and 62, optionally wherein the DR IL-18 polypeptide consists of, or consists essentially of, an amino acid sequence selected from SEQ ID NOs: 20, 7, 17-19, 21, 22, 55-57, and 62.

5. The method of any one of the preceding claims, wherein the amount of the engineered IL-18 polypeptide administered is from 30 pg / kg to 1200 pg / kg, optionally from 60 pg / kg to 240 Pg / kg-6. The method of any of the preceding claims, wherein the amount is administered as a single IV infusion.

7. The method of any of the preceding claims, wherein administering the effective amount of the engineered IL- 18 polypeptide comprises achieving a maximum concentration observed (Cmax) of at least 1000 ng / mL of the engineered IL- 18 polypeptide in plasma of the subject within 1 day of the administration, optionally within 12 hours of the administration, optionally within 6 hours of the administration.

8. The method of any of the preceding claims, wherein the method comprises repeating the administration weekly, once every two weeks, or once every three weeks, optionally wherein administration is repeated for at least four weeks or at least seven weeks.

9. The method of any one of the preceding claims, wherein the administering results in an engineered IL- 18 polypeptide area under the curve (AUC) accumulation ratio (AR) that is at least five times higher than the engineered IL- 18 polypeptide AUC AR resulting from a corresponding subcutaneous route of administration, an engineered IL- 18 polypeptide Cmax AR that is at least five times higher than the engineered IL- 18 polypeptide Cmax AR resulting from a corresponding subcutaneous route of administration, or both.

10. The method of any one of the preceding claims, wherein the subject has, following the administering, one or more reduced risks selected from the group consisting of: a reduced risk of developing an injection site reaction (ISR); a reduced risk of developing one or more symptoms of an ISR; a reduced risk of developing a Grade 3 ISR; or any combination thereof, as compared to a corresponding subject administered an equivalent amount of the engineered IL- 18 polypeptide delivered subcutaneously, intramuscularly, or intradermally.

11. The method of any one of the preceding claims, wherein the subject experiences fewer injection site reaction (ISR) symptoms, less severe ISR symptoms, or both, as compared to delivery of the engineered IL- 18 polypeptide subcutaneously, optionally wherein the ISR symptoms are selected from the group consisting of ulcers, necrosis, tissue damage, pain, lipodystrophy, edema, phlebitis, tenderness, erythema, itching, or any combination thereof.

12. The method of any one of the preceding claims, wherein the subject does not develop a Grade 3 ISR, does not develop a Grade 2 or higher ISR, or does not develop an ISR.

13. The method of any of the preceding claims, wherein the subject has a cancer and the method comprises administering the engineered IL- 18 polypeptide to treat the subject for the cancer, optionally wherein the cancer comprises a solid tumor, optionally wherein the cancer is selected from the group consisting of melanoma, Merkel cell carcinoma, renal cell carcinoma, urothelial cancer, triple negative breast cancer, non-small cell lung cancer, squamous cell carcinoma, head and neck cancer, microsatellite instability-high tumors, high tumor mutationburden cancer, mismatch repair deficient cancer, gastric cancer, cervical cancer, endometrial cancer, small cell lung cancer, esophageal cancer, hepatocellular carcinoma, and platinum resistant ovarian cancer, optionally wherein the cancer is associated with loss of Major Histocompatibility Complex (MHC) class I surface expression, optionally wherein the cancer is resistant to checkpoint inhibitor treatment, optionally wherein the cancer comprises an anti-PDl- resistant tumor.

14. The method of one any of the preceding claims, further comprising pre-treating the subject with an effective amount of a B-cell depleter prior to administering the amount of the engineered IL-18 polypeptide, optionally wherein the B-cell depleter comprises an anti-CD20 antibody, optionally wherein the anti-CD20 antibody is obinutuzumab.

15. A method of administering an engineered interleukin 18 (IL-18) polypeptide to a subject in need thereof, the method comprising: pre-treating the subject with an effective amount of a B-cell depleter to deplete B-cells in the subject; and administering the engineered IL-18 polypeptide to the B-cell depleted subject, optionally wherein the engineered IL- 18 polypeptide is a decoy-resistant (DR) IL- 18 polypeptide, optionally wherein the B-cell depleter is a B-cell depleting antibody, optionally an anti-CD20 antibody, optionally obinutuzumab.

16. The method of claim 15, wherein the administering comprises intravenous administration of the engineered IL- 18 polypeptide, optionally wherein the engineered IL- 18 polypeptide is administered at a dose of at least 1 pg / kg, at least 10 pg / kg, at least 20 pg / kg, at least 30 pg / kg, at least 60 pg / kg, at least 120 pg / kg, or at least 240 pg / kg.

17. The method of claim 15 or 16, wherein the subject has a cancer and the method comprises administering the engineered IL-18 polypeptide to treat the subject for the cancer, optionally wherein the cancer is a solid tumor.

18. The method of any one of the preceding claims, further comprising administering CRS prophylaxis to the subject prior to, or while, administering the engineered IL- 18 polypeptide.

19. Use of a composition comprising an engineered IL-18 polypeptide, in a method of treating a subject, according to any one of the preceding claims, wherein the subject is a human subject having cancer and the method treats the subject for the cancer.

20. A kit comprising: a pharmaceutical composition formulated for intravenous (IV) delivery to a subject, the pharmaceutical composition comprising an engineered IL- 18 polypeptide and a pharmaceutically acceptable carrier; andone or more IV delivery devices, optionally wherein the one or more IV delivery devices are selected from the group consisting of a syringe suitable for use with a syringe pump, an IV bag, an IV cannula, an IV catheter, and any combination thereof.