Interleukin-18 mimetics

Abstract

The present disclosure provides interleukin-18 mimetics comprising an amino acid sequence with less than 76% identity to mature human IL-18 having the amino acid sequence set forth in SEQ ID NO:11 which mimetics bind to human IL-18 receptor. These IL-18 mimetics induce IL-18 signaling activity. The IL-18 mimetics have advantageous properties as compared to the mature human IL-18, e.g., higher solubility, higher thermal stability, etc.

Description

Atty Dkt. No.: MOBI-016WOINTERLEUKIN-18 MIMETICSCROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 735,803 filed on December 18, 2024, which application is herein incorporated by reference in its entirety.INCORPORATION BY REFERENCE OF XML SEQUENCE LISTING

[0002] A Sequence Listing is provided herewith as a Sequence Listing XML, "MOBI- 016WO_SEQ._LIST.XML," created on December 17, 2025 and having a size of 27,654 bytes. The contents of the text file are incorporated by reference herein in their entirety.INTRODUCTION

[0003] Interleukin 18 (IL-18) is a pro-inflammatory cytokine involved in host defense against infections and regulates the innate and acquired immune response. IL-18 can stimulate T cells, NK cells, and myeloid cells. IL-18 is used extensively in research. IL-18 has been proposed as an immunotherapeutic agent for the treatment of cancer, given its ability to stimulate anti-tumor immune cells. However, IL-18 is difficult to express in soluble form.

[0004] Thus, there is a need for IL-18 like molecules. The present disclosure addresses these and other needs.SUMMARY

[0005] The present disclosure provides an IL-18 mimetic polypeptide that has IL-18 activity. For example, the IL-18 mimetic polypeptides of the present disclosure bind to IL-18 receptor (IL-18R). The IL- 18 mimetic polypeptides of the present disclosure can bind to IL-18R with an affinity comparable to or higher than wild-type IL-18 (e.g., recombinant human IL-18). The IL-18 mimetic polypeptides of the present disclosure can bind to IL-18R and induce IL-18 signaling activity. The IL-18 mimetic polypeptides of the present disclosure can induce IL-18 signaling activity to a level comparable to or higher than wildtype IL-18 (e.g., recombinant human IL-18).

[0006] An IL-18 mimetic polypeptide provided herein may have an amino acid sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the amino acid sequence set forth in any one of SEQ ID NOs: 1-Atty Dkt. No.: MOBI-016WO

[0007] An IL-18 mimetic polypeptide provided herein has one or more improved properties as compared to a recombinant IL-18 comprising a native IL-18 sequence, e.g., an IL-18 having the amino acid sequence set forth in SEQ ID NO:11. For example, the IL-18 mimetic polypeptide has higher solubility, thermal stability, and / or affinity for I L-18R as compared to recombinant IL-18 comprising a native IL-18 sequence.

[0008] Also provided herein are methods for using the disclosed IL-18 mimetic polypeptides. The methods include both non-therapeutic and therapeutic use.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] FIG. 1. Activity of IL-18 mimetics, MBIO-7501 and MBIO-7505 and recombinant human IL-18 (rhlL-18).

[0010] FIGS. 2A-2C provide an alignment of the amino acid sequence set forth in SEQ ID NOs: 1- 10, 21-24, and 26-27. 7504=SEQ ID NO:5; 7505=SEQ ID NO:2; 8013= SEQ ID NO: 26; 8014= SEQ ID NO: 27; 7506=SEQ ID NO:6; 7510=SEQ ID NO:10; 7509=SEQ ID NO:9; 7507=SEQ ID NO:7; 7508=SEQ ID NO:8; 7499=SEQ ID NO:3; 7500=SEQ ID NO:4; 7501=SEQ ID NO:1; 8009=SEQ ID NO: 21; 8011=SEQ ID NO: 23; 8010= SEQ ID NO: 22; and 8012= SEQ ID NO: 24.

[0011] FIG. 3A provides an alignment of the amino acid sequence of an IL-18 mimetic derived from MBIO-7501. This IL-18 mimetic derived from MBIO-7501 shows reduced binding IL-18 BP as compared to the binding to IL-18BP by MBIO-7501. 7501=SEQ ID NO:1; 8009=SEQ ID NO: 21; 8011=SEQ ID NO: 23; 8010= SEQ ID NO: 22; and 8012= SEQ ID NO: 24. Positions 52, 54, 57, 58, 61, 106, 111, and / or 112 numbered relative to SEQ ID NO:1 are marked with arrows to show that the IL1-8 mimetics 8009- 8012 have substitutions relative to MBIO-7501.

[0012] FIG. 3B shows the percent identity between 7501=SEQ ID NO:1; 8009=SEQ ID NO: 21; 8011=SEQ ID NO: 23; 8010= SEQ ID NO: 22; and 8012= SEQ ID NO: 24.

[0013] FIGS. 4A-4D provide an alignment of amino acid sequences of IL-18 mimetics, 8013 and 8014 derived from MBIO-7505. These IL-18 mimetics show reduced binding IL-18BP as compared to the binding to IL-18BP by MBIO-7505. Positions 52, 54, 57, 58, 61, 106, 111, and / or 112 numbered relative to SEQ ID NO:1 are marked with arrows to show the positions substituted in MBIO-7505 and the corresponding positions in MBIO-7504, MBIO-7505, MBIO-7506, MBIO-7507, MBIO-7508, MBIO-7509, and MBIO-7510 that can be substituted to generate an IL-18 mimetic that does not significantly bind to IL-18BP. 7504=SEQ ID NO:5; 7505=SEQ ID NO:2; 8013=SEQ ID NO:27; 8014=SEQ ID NO:28; 7506=SEQ ID NO:6; 7510=SEQ ID NO:10; 7509=SEQ ID NO:9; 7507=SEQ ID NO:7; 7508=SEQ ID NO:8.Atty Dkt. No.: MOBI-016WO

[0014] FIGS. 5A-5C. Representative binding sensorgrams of wild-type IL-18 and IL-18 mimetics to IL-18 binding protein.

[0015] FIGS. 6A-6B. Representative binding sensorgrams of IL-18 mimetic MBIO-8013 to IL-18 receptor and IL-18 binding protein.

[0016] FIGS. 7A-7B. Biological activity of IL-18 mimetics in T-cell activation assays.

[0017] FIG. 8. Thermal stability of IL-18 mimetics assessed by biolayer interferometry.

[0018] FIGS. 9A-9B. Long-term thermal stability and retention of activity of IL-18 mimetics in cell-based assays.DETAILED DESCRIPTION

[0019] The present disclosure provides an IL-18 mimetic polypeptide that has IL-18 activity. For example, the IL-18 mimetic polypeptides of the present disclosure bind to IL-18 receptor (IL-18R). The IL- 18 mimetic polypeptides of the present disclosure can bind to IL-18R and induce IL-18 signaling activity.

[0020] 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.

[0021] 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 may 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.

[0022] 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 may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number.Atty Dkt. No.: MOBI-016WO

[0023] 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.

[0024] 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 prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0025] 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.

[0026] 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.

[0027] 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.Atty Dkt. No.: MOBI-016WODEFINITIONS

[0028] "Derived from" in the context of an amino acid sequence or polynucleotide sequence is meant to indicate that the polypeptide or nucleic acid has a sequence that is based on that of a reference polypeptide or nucleic acid, and is not meant to be limiting as to the source or method in which the protein or nucleic acid is made.

[0029] The terms "polypeptide", and "protein" are used interchangeably herein to designate a linear series of amino acid residues connected one to the other by peptide bonds between the alphaamino and carboxy groups of adjacent residues. The amino acid residues are usually in the natural "L" isomeric form. However, residues in the "D" isomeric form can be substituted for any L-amino acid residue, as long as the desired functional property is retained by the polypeptide. In addition, the amino acids, in addition to the 20 "standard" amino acids, include modified and unusual amino acids, which include, but are not limited to those listed in 37 CFR (§1.822(b)(4)). The term "peptide" also refers to a linear series of amino acid residues connected one to the other by peptide bonds between the alphaamino and carboxy groups of adjacent residues but is generally shorter than a protein or a polypeptide, e.g., less than 50 amino acids long, e.g., 2-50 amino acids in length. The terms protein, polypeptide, and peptide may be used interchangeably.

[0030] As used herein, the term "binding" refers to the non-covalent interactions of the type which occur between two molecules. The strength or affinity of binding interactions can be expressed in terms of the dissociation constant (KD) of the interaction, wherein a smaller KDrepresents a greater affinity. Binding properties of selected polypeptides can be quantified using methods well known in the art.

[0031] "Isolated" refers to an entity of interest that is in an environment different from that in which the entity may naturally occur or is initially produced in. An "isolated" compound (e.g., an "isolated" polypeptide) is separated from all or some of the components that accompany it and may be substantially enriched, e.g., may be purified so that the compound is at least about 70% pure, at least about 80% pure, at least about 90% pure, at least about 95% pure, at least about 98% pure, at least about 99%, or greater than 99% pure, or free of impurities, contaminants, and / or components other than the compound. "Isolated" also refers to the state of a compound separated from all or some of the components that accompany it during manufacture (e.g., chemical synthesis, recombinant expression, culture medium, and the like).

[0032] As used herein, the amino acid residues are abbreviated as follows: alanine (Ala; A), asparagine (Asn; N), aspartic acid (Asp; D), arginine (Arg; R), cysteine (Cys; C), glutamic acid (Glu; E),Atty Dkt. No.: MOBI-016WO glutamine (Gin; Q), glycine (Gly; G), histidine (His; H), isoleucine (lie; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Vai; V).

[0033] In all embodiments of polypeptides disclosed herein, any N-terminal methionine residues are optional (i.e., the N-terminal methionine residue may be present or absent). In some embodiments, one or more of the amino acids may have a modification, such as, a post-translational modification, e.g., acetylation or glycosylation.

[0034] The term "conservative substitution" is used in reference to proteins to reflect amino acid substitutions that do not substantially alter the activity (specificity or binding affinity) of the molecule. Typically, conservative amino acid substitutions involve substituting one amino acid for another amino acid with similar chemical properties (e.g., charge or hydrophobicity). The following six groups each contain amino acids that are typical conservative substitutions for one another: 1) Alanine (A), Serine (S), Threonine (T); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W). The polypeptides encompassed by the present disclosure include those that have one or more conservative substitutions relative to the amino acid sequences provided here.

[0035] Percent identity between a pair of sequences may be calculated by multiplying the number of matches in the pair by 100 and dividing by the length of the aligned region, including gaps. Identity scoring only counts perfect matches and does not consider the degree of similarity of amino acids to one another. Unless specified otherwise, only internal gaps are included in the length, not gaps outside of the aligned regions. Percent Identity = (Matches x 100) / Length of aligned region (with gaps).

[0036] Percent identity between a sequence and the entire length of a reference sequence may be calculated by multiplying the number of matches in the two sequences by 100 and dividing by the entire length of the reference sequence, including internal gaps and gaps outside of the aligned regions. Percent Identity between a sequence and the entire length of a reference sequence = (Matches x 100) / Length of the reference sequence.

[0037] Numeric ranges are inclusive of the numbers defining the range.

[0038] The terms "patient," "subject," "individual," and the like are used interchangeably herein, and refer to any mammal, or cells thereof whether in vivo, in vitro or in situ, amenable to the methods described herein. In certain non-limiting embodiments, the patient, subject or individual is a human.Atty Dkt. No.: MOBI-016WO

[0039] The term "therapeutically effective amount" refers to the amount of the subject polypeptide, nucleic acid, cells or composition that will elicit the biological, physiological, clinical or medical response of a cell, tissue, organ, system, or subject that is being sought by the researcher, veterinarian, medical doctor or other clinician. The term "therapeutically effective amount" includes that amount that, when administered, is sufficient to prevent development of, or ameliorate, one or more of the signs or symptoms of the disorder or disease being treated. The therapeutically effective amount will vary depending on the compound or composition, the disease and its severity and the age, weight, etc., of the subject to be treated.

[0040] To "treat" a disease or disorder as the term is used herein, means to reduce the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject. The terms "treatment", "treating", "treat" and the like are used herein to generally refer to obtaining a desired pharmacologic and / or physiologic effect. The effect can be prophylactic in terms of completely or partially preventing a disease or symptom(s) thereof and / or may be therapeutic in terms of a partial or complete stabilization or cure for a disease and / or adverse effect attributable to the disease. The term "treatment" encompasses any treatment of a disease in a mammal, particularly a human, and includes: (a) preventing the disease and / or symptom(s) from occurring in a subject who may be predisposed to the disease or symptom but has not yet been diagnosed as having it; (b) inhibiting the disease and / or symptom(s), e.g., slowing or arresting their development (e.g., halting the growth of tumors, slowing the rate of tumor growth, halting the rate of cancer cell proliferation, and the like); or (c) relieving the disease symptom(s), i.e., causing regression of the disease and / or symptom(s) (e.g., causing decrease in tumor size, reducing the number of cancer cells present, and the like). Those in need of treatment include those already inflicted (e.g., those with cancer, those with an infection, those with a metabolic disorder (e.g., obesity and diabetes and the like), etc.) as well as those in which prevention is desired (e.g., those with increased susceptibility to cancer, those with an increased likelihood of infection, those suspected of having cancer, those suspected of harboring an infection, those with increased susceptibility for metabolic disease(e.g., obesity and diabetes and the like), etc.).

[0041] As used herein, the term "wild-type" refers to a gene or gene product isolated from a naturally occurring source. A wild-type gene is that which is most frequently observed in a population and is thus arbitrarily designated the "normal" or "wild-type" form of the gene. In contrast, the term "modified," "variant," or "mutant" refers to a gene or gene product that possesses modifications in sequence and / or functional properties (i.e., altered characteristics) when compared to the wild-type gene or gene product.Atty Dkt. No.: MQBI-016WQIL-18 MIMETICS

[0042] The present disclosure provides IL-18 mimetic polypeptides that have IL-18 activity. For example, the IL-18 mimetic polypeptides of the present disclosure bind to IL-18 receptor (IL-18R). The IL- 18 mimetic polypeptides of the present disclosure can bind to IL-18R, e.g., a human IL-18R and induce IL- 18 signaling activity. The terms "IL-18 mimetic polypeptide" and "IL-18 mimetic" and "IL-18 mimic" and the terms "IL-18 mimetic polypeptides" and "IL-18 mimetics" and "IL-18 mimics" are used interchangeably.

[0043] An IL-18 mimetic polypeptide provided herein may include an amino acid sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the amino acid sequence set forth in any one of SEQ ID NOs: 1- 10 and 20-27. An IL-18 mimetic polypeptide provided herein may include an amino acid sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the amino acid sequence set forth in any one of SEQ ID NOs: 1- 10 and 20-27 and includes conservative amino acid substitutions relative to the sequence of any one of SEQ ID NOs: 1-10 and 20-27. FIG. 2 provides an alignment of the amino acid sequences set forth in SEQ ID NOs: 1-10, 21-24, and 26-27.

[0044] An IL-18 mimetic polypeptide provided herein may have a length of 120-160 amino acids, e.g., 130-160, 140-160, 150-160, 155-160, or 150-158 amino acids. In addition, the IL-18 mimetic polypeptide may include a Methionine at the N-terminus and / or at the N or the C terminus one or more of amino acids introduced via a cloning vector, signal peptide, purification tags, detection tags, and linkers, and the like. In certain aspects, the IL-18 mimetic polypeptide provided herein do not include a cysteine residue.

[0045] An IL-18 mimetic polypeptide provided herein has one or more improved properties as compared to a recombinant IL-18 comprising a native IL-18 sequence, e.g., a mature IL-18 having the amino acid sequence set forth in SEQ ID NO:11 (UniProt Accession No. Q14116). For example, the IL-18 mimetic polypeptide has higher solubility, thermal stability, and / or affinity for IL-18R as compared to recombinant IL-18 comprising a native IL-18 sequence of SEQ ID NO:11.

[0046] As used herein, the term "solubility" refers to a property of a polypeptide that allows the polypeptide to remain in solution instead of precipitating. Soluble protein maintains correct folding (e.g., native structure) and retains activity while an insoluble protein can form aggregates and has no or low activity. IL-18 mimetics of the present disclosure are more soluble when expressed recombinantlyAtty Dkt. No.: MQBI-016WQ(e.g., from a genetically modified prokaryotic or eukaryotic cell) as compared to recombinantly expressed IL-18 polypeptide having the amino acid sequence set forth in SEQ ID NO:11. Due to the higher solubility, the recombinantly expressed IL-18 mimetics of the present disclosure have a higher yield (e.g., at least 5%, at least 10%, at least 15%, at least 25%, at least 40%, or at least 50% higher) as compared to the recombinantly expressed IL-18 polypeptide having the amino acid sequence set forth in SEQ ID NO:11.

[0047] As used herein, the term "thermal stability" refers to ability of a polypeptide to maintain native conformation and function when exposed to elevated temperatures, e.g., a temperature higher than 37°C (such as, higher than 40°C, higher than 50°C, higher than 60°C, higher than 70°C, higher than 80°C, and up to 100°C).

[0048] The higher thermal stability of IL-18 mimetics polypeptides of the present disclosure can increase yield of the recombinantly produced IL-18 mimetic polypeptides. The higher thermal stability of IL-18 mimetics polypeptides is advantageous during cell culture enabling use of the IL-18 mimetics polypeptides during extended cell culture duration, e.g., the IL-18 mimetics polypeptides retain activity for at least 1 day, at least 3 days, at least 1 week, at least 10 days, at least 2 weeks or longer, e.g., up to 3 weeks.

[0049] The higher thermal stability of the IL-18 mimetics polypeptides provides more consistent activity in inducing cell differentiation, cell expansion, and manufacture of differentiated cells as compared to rhlL-18 having the amino acid sequence set forth in SEQ ID NO:11.

[0050] Thermal stability may be measured using any standard assay. Examples include subjecting the protein to a series of high temperatures and measuring activity and / or solubility.

[0051] The IL-18 mimetic polypeptides disclosed herein may bind to IL-18 binding protein (IL- 18BP), e.g., human IL-18BP (hl L-18BP). The IL-18 mimetic polypeptides disclosed herein may bind to hlL- 18BP with an affinity similar to that of rhlL-18. Examples of IL-18 mimetic polypeptides that bind to hlL- 18BP with an affinity similar to that of rhlL-18 include IL-18 mimetic polypeptides comprising an amino acid sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the amino acid sequence set forth in any one of SEQ ID NOs: 1-10. IL-18 mimetics that bind to IL-18BP comprise an amino acid sequence having at least 80% identity (e.g., at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO:1 and comprise (i) L 52; (ii) K 54; (iii) Q 57; (iv) P 58; (v) M 61; (vi) S 106; (vii) D 111; and / or (viii) N 112, wherein the numbering of the amino acids is based on the amino acid sequence set forth in SEQ ID NO:1 (MBIQ-7501). In another example, IL-18 mimetics that bind to IL-Atty Dkt. No.: MQBI-016WO18BP comprise an amino acid sequence having at least 80% identity (e.g., at least 85%, at least 90%, at least 95%, at least 95%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO:2 and comprise (i) M 52; (ii) K 54; (iii) Q 57; (iv) P 58; (v) M 61; (vi) S 106; (vii) D 111; and / or (viii) N 112, wherein the numbering of the amino acids is based on the amino acid sequence set forth in SEQ ID NO:1 (MBIO- 7501).

[0052] The IL-18 mimetic polypeptides disclosed herein may bind to hlL-18BP with an affinity lower to that of rhlL-18. The IL-18 mimetic polypeptides disclosed herein may not show significant binding to rhl L-18BP. The IL-18 mimetic polypeptides disclosed herein do not show significant binding to hl L-18BP and are referred to as a decoy-resistant IL-18 mimetic. The IL-18 mimetic that bind to IL-18R and exhibit substantially reduced binding to IL-18BP has an I L-18BP / I L-18R dissociation constant ratio that is about at least 2-fold higher (e.g., at least 20-fold higher or at least 200-fold higher) than the IL- 18BP / IL-18R dissociation constant ratio of wild-type IL-18 (an increased dissociation constant ratio indicates a relative decrease in IL-18BP binding relative to IL- 18R binding). IL-18 has immunostimulatory effects but is negatively regulated by IL-18BP that limits IL-18's anticancer efficacy. A decoy-resistant IL- 18 mimetic avoids sequestration by IL-18BP while maintaining its immunostimulatory activity. The decoy-resistant IL-18 mimetics of the present disclosure may be derived from the IL-18 mimetics disclosed herein and may have 1, 2, 3, 4, 5, 6, 7, or 8 substitutions relative to the amino acid sequence of the IL-18 mimetics that show significant binding to IL-18BP. These amino acid substitution(s) may be at the following positions:

[0053] (i) 52; (ii) 54; (iii) 57; (iv) 58; (v) 61; (vi) 106; (vii) 111; and / or (viii) 112, wherein the numbering of the amino acids being substituted is based on the amino acid sequence set forth in SEQ ID NO:1 (MBIO-7501).

[0054] A decoy-resistant IL-18 mimetic of the present disclosure may include 1, 2, 3, 4, 5, 6, 7, or 8 of the following substitutions:

[0055] (i) L 52 -> K; (ii) K 54 -> S; (iii) Q 57 -> L; (iv) P 58 -> A; (v) M 61 -> L; (vi) S 106 -> D; (vii) D111 -> S; (viii) N 112 -> R relative to SEQ ID NO:1, wherein the numbering of the amino acids being substituted is based on the amino acid sequence set forth in SEQ ID NO:1 (MBIO-7501).

[0056] A decoy-resistant IL-18 mimetic of the present disclosure may have an amino acid sequence at least 80% identical (e.g., at least 85%, at least 90%, or at least 95% identical) to the amino acid sequence of any one of SEQ ID NQs:l-10 and 20-27 and include 1, 2, 3, 4, 5, 6, 7, or 8 of the following substitutions: (i) 52 -> K; (ii) 54 -> S; (iii) 57 -> L; (iv) 58 -> A; (v) 61 -> L; (vi) 106 -> D; (vii) 111 -> S; and (viii) 112 -> R, wherein the numbering of the amino acids being substituted is based on the aminoAtty Dkt. No.: MQBI-016WQ acid sequence set forth in SEQ ID NO:1 (MBIO-7501) and the substitutions are relative to the amino acid sequence of any one of SEQ ID NOs:l-10 and 20-27.

[0057] A decoy-resistant IL-18 mimetic of the present disclosure may have an amino acid sequence at least 80% identical (e.g., at least 85%, at least 90%, or at least 95% identical) to the amino acid sequence of SEQ ID NO:1 and include 1, 2, 3, 4, 5, 6, 7, or 8 of the following substitutions: (i) L 52 -> K; (ii) K 54 -> S; (iii) Q 57 -> L; (iv) P 58 -> A; (v) M 61 -> L; (vi) S 106 -> D; (vii) D 111 -> S; (viii) N 112 -> R, relative to SEQ ID NO:1, wherein the numbering of the amino acids being substituted is based on the amino acid sequence set forth in SEQ ID NO:1 (MBIO-7501).

[0058] A decoy-resistant IL-18 mimetic of the present disclosure may have an amino acid sequence at least 80% identical (e.g., at least 85%, at least 90%, or at least 95% identical) to the amino acid sequence of SEQ ID NO:2 and include 1, 2, 3, 4, 5, 6, 7, or 8 of the following substitutions: (i) M 52 -> K; (ii) K 54 -> S; (iii) Q 57 -> L; (iv) P 58 -> A; (v) M 61 -> L; (vi) S 106 -> D; (vii) D 111 -> S; (viii) N 112 -> R, relative to SEQ ID NO:2, wherein the numbering of the amino acids being substituted is based on the amino acid sequence set forth in SEQ ID NO:1 (MBIO-7501).

[0059] Human IL-18 is expressed as a biologically inactive precursor, pro-IL-18, which is cleaved by caspase-1 (IL-ip-converting enzyme, ICE) to form the biologically active mature cytokine. Direct expression of mature recombinant human IL-18 in E. coli results in a partially active cytokine. For human IL-18 (HulL-18), it has been shown that correct folding of hulL-18 requires its prior synthesis as pro-IL-18 (Liu, B., Cytokine, Volume 12, Issue 10, 2000, Pages 1519-1525). An IL-18 mimetic polypeptide of the present disclosure may be expressed as a pro-IL-18- mimetic polypeptide which is cleaved by caspase-1 to generate an active IL-18 mimetic polypeptide. An IL-18 mimetic polypeptide of the present disclosure may not be expressed as pro-IL-18 and instead is expressed directly as an active IL-18.

[0060] The IL-18 mimetics disclosed herein may include a cleavable signal peptide at the N- or the C-terminus. The IL-18 mimetics disclosed herein may include additional amino acids at the N- or the C-terminus, e.g., an affinity tag, such as, a 6X His-tag.

[0061] An IL-18 mimetic polypeptide of the present disclosure may not have the amino acid sequence of any one of SEQ ID NOs: 12-19:Atty Dkt. No.: MOBI-016WOAtty Dkt. No.: MOBI-016WO

[0062] An IL-18 mimetic polypeptide disclosed herein may be in a pharmaceutical composition comprising the IL-18 mimetic and a pharmaceutically acceptable excipient. The pharmaceutical composition may further include:

[0063] an immune checkpoint inhibitor that inhibits PD-L1, PD1, CTLA4, TIM3, TIGIT, LAG3, B7H3, B7H4, VISTA, BTLA, CD47, SIRP alpha, CD48, CD155, CD160, TREM2, IDO1, Adenosine 2A receptor, Aryl hydrocarbon receptor, KIR, LILRB2, or any combination thereof;

[0064] an immune agonist selected from: an agent that agonizes a tumor necrosis factor receptor superfamily (TNFRSF) protein (e.g., GITR, 4 IBB, 0X40, CD27, CD40, HVEM); an agent that agonizes an immunoglobulin superfamily (IgSF) protein (e.g., CD28, ICOS, CD226, NKG2D); an agent that agonizes a Toll-like Receptor (TLR) (e.g., TLR2, TLR4, TLR5, TLR7, TLR9); an agent that agonizes a nucleic acid sensor (e.g., STING, cGAS, RIG-1 (DDX58)); an inflammasome activator; a T cell engager; a cytokine or cytokine variant (e.g., IL-2, IL-7, IL-10, IL-12, IL-15, IL-21, IL-23, IL-27, IL-33, TNF,TL1 A,IFNA,IFNB,IFNG); or any combination thereof;

[0065] a cancer cell opsonizing agent that targets one or more antigens selected from: CD19, CD20, CD22, CD24, CD25, CD30, CD33, CD37, CD38, CD44, CD45, CD47, CD51, CD52, CD56, CD62L, CD70, CD74, CD79, CD80, CD96, CD97, CD99, CD123, CD134, CD138, CD152 (CTLA-4), CD200, CD213A2, CD221, CD248, CD276 (B7-H3), B7-H4, CD279 (PD-1), CD274 (PD-L1), CD319, EGFR, EPC AM, 17-1 A, HER1, HER2, HER3, CD 117, C-Met, HGFR, PDGFRA, AXL, TWEAKR, PTHR2, HAVCR2 (TIM3), GD2 ganglioside, MUC1, mucin CanAg, mesothelin, endoglin, Lewis-Y antigen, CEA, CEACAM1, CEACAM5, CA-125, PSMA, BAFF, FGFR2, TAG-72, gelatinase B, glypican 3, nectin-4, BCMA, CSF1R, SLAMF7, integrin av 3, TYRP1, GPNMB, CLDN18.2, F0LR1, CCR4, CXCR4, MICA, C242 antigen, DLL3, DLL4, EGFL7, vimentin, fibronectin extra domain-B, TROP-2, LRRC15, FAP, SLITRK6, NOTCH2, NOTCH3, Tenascin-3, STEAP1, and NRPI; or

[0066] an engineered T or NK cell, a chimeric antigen receptor T cell (CAR-T cell), an engineered TCR-T cell, a chimeric antigen receptor NK cell (CAR-NK cell), or a Tumor-infiltrating Lymphocyte (TIL).

[0067] A composition comprising an IL-18 mimetic as described herein may comprise more than one such IL-18 mimetic. For example, the composition may comprise two or more different IL-18 mimetics. The composition may comprise an IL-18 mimetic that binds to IL-18BP and another IL-18 mimetic that does not bind to IL-18BP, for example.

[0068] Also provided herein are methods for using the disclosed IL-18 mimetic polypeptides. The methods include both non-therapeutic and therapeutic use.Atty Dkt. No.: MOBI-016WOConjugated Polypeptides

[0069] The IL-18 mimetic polypeptides disclosed herein may be conjugated to another moiety. The moiety may be a small molecule, peptide, polypeptide, nucleic acid, or lipid. The polypeptides may also be tagged with a sequence for localization to a cellular compartment, cell membrane, for secretion, detection, or purification.

[0070] The polypeptides disclosed herein may be labeled by conjugating a detectable moiety to the N-terminus, the C-terminus, or between the N- and the C-terminus. The detectable moiety may be a radioisotope, an enzyme which generates a detectable product, a fluorescent protein, and the like.

[0071] The moiety may be conjugated directly to the polypeptides, e.g., via a peptide bond to the N-terminus and / or the C-terminus and / or to an amino acid side chain or may be conjugated to the polypeptide via a linker. The linker may be a polymer, e.g., an amino acid linker or a sugar linker. The moiety may be conjugated to the side chain of the amino acid at the N-terminus and / or the side chain of the amino acid at the C-terminus and / or to an interior amino acid side chain of the polypeptides disclosed herein. In certain examples, an amino acid, e.g., a cysteine or a lysine may be introduced at the N-terminus and / or the C-terminus and / or in between the N- and C-terminus of the polypeptides disclosed herein to serve as a chemical handle for conjugating a moiety.

[0072] A variety of linkers may be used and may include alkyl groups, methylene carbon chains, ether, polyether, alkyl amide linker, a peptide linker, a modified peptide linker, a Polyethylene glycol) (PEG) linker, a streptavidin-biotin or avidin-biotin linker, polyaminoacids (e.g., polylysine), functionalized PEG, polysaccharides, glycosaminoglycans, oligonucleotide linker, phospholipid derivatives, alkenyl chains, alkynyl chains, disulfide, or a combination thereof. In some embodiments, the linker is cleavable (e.g., enzymatically (e.g., Tobacco etch virus (TEV) protease site), chemically, photoinduced cleavage, etc.).

[0073] In certain aspects, the moiety may be a heterologous amino acid sequence. In certain aspects, the moiety is conjugated to the polypeptide post-translationally. In certain aspects, the moiety is conjugated to the polypeptide during translation, e. g., a nucleic acid may encode a fusion protein comprising the polypeptide and the moiety. As used herein, the term "heterologous amino acid sequence" means an amino acid sequence that is not part of a polypeptide's sequence prior to the conjugation or fusion. Heterologous amino sequences of interest include amino acid sequences of a polypeptide that interact with another polypeptide either directly or via another moiety.

[0074] In certain aspects, the heterologous amino acid sequence includes a protein binding domain, such as one that binds IL-17RA, e.g., IL-17A, or the IL-17A binding domain of IL-17RA, JunAtty Dkt. No.: MOBI-016WO binding domain of Erg, or the EG binding domain of Jun; a potassium channel voltage sensing domain, e.g., one useful to detect protein conformational changes, the GTPase binding domain of a Cdc42 or rac target, or other GTPase binding domains, domains associated with kinase or phosphatase activity, e.g., regulatory myosin light chain, PKC6, pleckstrin containing PH and DEP domains, other phosphorylation recognition domains and substrates; glucose binding protein domains, glutamate / aspartate binding protein domains, PKA or a cAMP-dependent binding substrate, lnsP3 receptors, GKI, PDE, estrogen receptor ligand binding domains, apoKl-er, or calmodulin binding domains.

[0075] In certain aspects, a fusion protein comprising a polypeptide fused to a heterologous amino acid sequence may be a biosensor. The biosensor is useful to detect a GTPase, e.g., binding of Cdc42 or Rac to a EBFP, EGFP PAK fragment, Raichu-Rac, Raichu-Cdc42, integrin alphavbeta3, IBB of importin-a, DMCA or NBD-Ras of CRafl (for Ras activation), binding domain of Ras / Rap Rai RBD with Ras prenylation sequence. In one embodiment, the biosensor detects PI(4,5)P2 (e.g., using PH-PCLdeltal, PH-GRP1), PI(4,5)P2 or PI(4)P (e.g., PH-OSBP), PI(3,4,5)P3 (e.g., using PH-ARNO, or PH-BTK, or PH- Cytohesinl), PI(3,4,5)P3 or PI(3,4)P2 (e.g., using PH Akt), PI(3)P (e.g., using FYVE-EEA1), or Ca2+ (cytosolic) (e.g., using calmodulin, or C2 domain of PKC.

[0076] In one aspect, a fusion protein comprising a polypeptide is fused to a protein domain. In one embodiment, the domain is one with a phosphorylated tyrosine (e.g., in Src, Abl and EGFR), that detects phosphorylation of ErbB2, phosphorylation of tyrosine in Src, Abl and EGFR, activation of MKA2 (e.g., using MK2), cAMP induced phosphorylation, activation of PKA, e.g., using KID of CREG, phosphorylation of Crkll, e.g., using SH2 domain pTyr peptide, binding of bZIP transcription factors and REL proteins, e.g., bFos and bJun ATF2 and Jun, or p65 NFkappaB, or microtubule binding, e.g., using kinesin.

[0077] The heterologous amino acid sequence fused to the polypeptides of the present disclosure may be an immunoglobulin G (IgG) Fc sequence, e.g., a human IgG Fc sequence, such as, human IgGl Fc sequence or a fluorescent protein.

[0078] The heterologous amino acid sequence fused to the polypeptides of the present disclosure may be a purification tag. The purification tag may be conjugated to the polypeptide via a cleavable or non-cleavable linker. The cleavable linker may be cleaved to produce a tag-free polypeptide after purification of the fusion protein using the tag. Appropriate affinity tags are known in the field and include poly-histidine tag (6X-H is), Glutathione S-Transferase (GST) tag, HA tag, Myc tag, FLAG tag, HSA tag, fluorescent tag, etc.Atty Dkt. No.: MOBI-016WO

[0079] The heterologous amino acid sequence fused to the polypeptides of the present disclosure may be fused to human serum albumin or to an antibody or a fragment thereof that binds to human serum albumin.

[0080] The heterologous amino acid sequence fused to the polypeptides of the present disclosure may be an interleukin, e.g., one or more of IL-12, IL-15, etc.

[0081] The IL-18 mimetic polypeptides may be used in an in vitro method for activation and proliferation of a natural killer cell or a T cell. The method may include contacting a natural killer cell or a T cell in a cell culture medium with an IL-18 mimetic polypeptide.NUCLEIC ACIDS

[0082] The present disclosure provides nucleic acids comprising nucleotide sequences encoding the polypeptides described herein. These nucleic acids may be used for a cell-free transcription and translation. A nucleotide sequence encoding a subject polypeptide can be operably linked to one or more regulatory elements, such as a promoter and enhancer, that allow expression of the nucleotide sequence in a recombinant cell that is genetically modified to produce the polypeptide.

[0083] Suitable promoter and enhancer elements are known in the art. For expression in a bacterial cell, suitable promoters include, but are not limited to, lacl, lacZ, T3, T7, gpt, lambda P and trc. For expression in a eukaryotic cell, suitable promoters include, but are not limited to, cytomegalovirus immediate early promoter; herpes simplex virus thymidine kinase promoter; early and late SV40 promoters; promoter present in long terminal repeats from a retrovirus; mouse metallothionein-l promoter; and the like.

[0084] A nucleotide sequence encoding a subject polypeptide can be present in an expression vector and / or a cloning vector. An expression vector can include a selectable marker, an origin of replication, and other features that provide for replication and / or maintenance of the vector. Large numbers of suitable vectors and promoters are known to those of skill in the art; many are commercially available for generating a subject recombinant construct. The following vectors are provided by way of example. Bacterial: pBs, phagescript, PsiX174, pBluescript SK, pBs KS, pNH8a, pNH16a, pNH18a, pNH46a (Stratagene, La Jolla, Calif., USA); pTrc99A, pKK223-3, pKK233-3, pDR540, and pRIT5 (Pharmacia, Uppsala, Sweden). Eukaryotic: pWLneo, pSV2cat, pOG44, PXR1, pSG (Stratagene) pSVK3, pBPV, pMSG and pSVL (Pharmacia). Expression vectors generally have convenient restriction sites located near the promoter sequence to provide for the insertion of nucleic acid sequences encoding polypeptides. A selectable marker operative in the expression host cell may be present.Atty Dkt. No.: MOBI-016WO

[0085] Nucleic acids, e.g., as described herein, may, in some instances, be introduced into a cell, e.g., by contacting the cell with the nucleic acid. Cells with introduced nucleic acids will generally be referred to herein as genetically modified cells. Various methods of nucleic acid delivery may be employed including, but not limited to e.g., naked nucleic acid delivery, viral delivery, chemical transfection, biolistics, and the like.

[0086] The nucleic acids of the present disclosure may be provided in a kit. The kit may include additional components such as reconstitution buffer for resuspending the nucleic acid provided in the kit in a lyophilized form. The nucleic acids of the present disclosure may be codon optimized to enhance expression in a particular cell type.GENETICALLY MODIFIED CELLS

[0087] The present disclosure provides isolated genetically modified cells (e.g., in vitro cells, ex vivo cells, cultured cells, etc.) that are genetically modified with a subject nucleic acid. In some aspects, a subject isolated genetically modified cell can produce a subject polypeptide. In some instances, a genetically modified cell may be used in the screening, and / or discovery of protein-protein interaction; protein-drug interactions; protein-nucleic acid interaction, etc.

[0088] Suitable cells include eukaryotic cells, such as a mammalian cell, an insect cell, a yeast cell; and prokaryotic cells, such as a bacterial cell. The bacterial cell may be an E. coli cell. Introduction of a subject nucleic acid into the host cell can be affected, for example by calcium phosphate precipitation, DEAE dextran mediated transfection, liposome-mediated transfection, electroporation, or other known methods.KITS

[0089] Aspects of the present disclosure include kits comprising an IL-18 mimetic polypeptide as disclosed herein. In one aspect, the kit comprises (i) the polypeptide as disclosed herein, (ii) the nucleic acid comprising a nucleotide sequence encoding the polypeptide as disclosed herein or the vector including the nucleic acid. The kit may include instructional material describing how to make and / or use the IL-18 mimetic polypeptide or the nucleic acid comprising a nucleotide sequence encoding the polypeptide. "Instructional material," as that term is used herein, includes a publication, a recording, a diagram, or any other medium of expression which can be used to communicate the usefulness of the nucleic acid or polypeptide of the disclosure in the kit for, e.g., identifying or alleviating or treating the various diseases or disorders recited herein. Optionally, or alternately, the instructional material mayAtty Dkt. No.: MOBI-016WO describe one or more methods of identifying or alleviating the diseases or disorders in a cell or a tissue of a subject. The instructional material of the kit may, for example, be affixed to a container that contains the nucleic acid or polypeptide or cells of the disclosure or be shipped together with a container that contains the nucleic acid or polypeptide or cells of the disclosure. Alternatively, the instructional material may be shipped separately from the container with the intention that the recipient uses the instructional material and the nucleic acid or polypeptide or cells of the disclosure cooperatively.COMPOSITIONS

[0090] The disclosure encompasses the preparation and use of pharmaceutical compositions comprising the IL-18 mimetic of the present disclosure, the nucleic acid of the present disclosure, or the cell of the present disclosure, useful for the treatment or prevention of a disease or disorder. Such a pharmaceutical composition may further comprise one or more pharmaceutically acceptable carriers, one or more additional ingredients, or some combination thereof.

[0091] In some embodiments, pharmaceutical compositions can include large, slowly metabolized macromolecules such as proteins, polysaccharides such as chitosan, polylactic acids, polyglycolic acids and copolymers (such as latex functionalized Sepharose™, agarose, cellulose, and the like), polymeric amino acids, amino acid copolymers, and lipid aggregates (such as oil droplets or liposomes).

[0092] The pharmaceutical compositions useful for practicing the invention may be administered to deliver a dose of between about 0.1 ng / kg / day and 100 mg / kg / day, or more.

[0093] In various embodiments, the pharmaceutical compositions useful in the methods of the disclosure may be administered, by way of example, systemically, parenterally, or topically, such as, in oral formulations, inhaled formulations, including solid or aerosol, and by topical or other similar formulations. In addition to the appropriate therapeutic composition, such pharmaceutical compositions may contain pharmaceutically acceptable carriers and other ingredients known to enhance and facilitate drug administration. Other possible formulations, such as nanoparticles, liposomes, other preparations containing the active ingredient, and immunologically based systems may also be used to administer an appropriate modulator thereof, according to the methods of the disclosure.

[0094] Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyidimethylbenzylAtty Dkt. No.: MOBI-016WO ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). Formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.

[0095] Compositions can be prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection can also be prepared. The preparation also can be emulsified or encapsulated in liposomes or micro particles such as polylactide, polyglycolide, or copolymer for enhanced adjuvant effect, as discussed above. Langer, Science 249: 1527, 1990 and Hanes, Advanced Drug Delivery Reviews 28: 97-119, 1997. The agents of this disclosure can be administered in the form of a depot injection or implant preparation which can be formulated in such a manner as to permit a sustained or pulsatile release of the active ingredient. The pharmaceutical compositions are generally formulated as sterile, substantially isotonic and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration.

[0096] The pharmaceutical compositions of the present disclosure may be formulated for injection, e.g., parenteral administration. As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue. Parenteral administration includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration includes, but is not limited to, cutaneous, subcutaneous, intraperitoneal, intravenous, intramuscular, intracisternal injection, and kidney dialytic infusion techniques.Atty Dkt. No.: MOBI-016WOMETHODS

[0097] The IL-18 mimetics disclosed herein can be used for all applications that the wild type IL- 18 is used for. In addition, the IL-18 mimetics that show significantly reduced binding to IL-18BP can be used in method where IL-18BP reduces the effectiveness of IL-18.

[0098] In some embodiments, IL-18 mimetics are used for the treatment and prevention of a disease or disorder. In various embodiments, the disease or disorder is cancer or an infectious disease, such as poxviruses that encode an IL-18BP ortholog, a metabolic disease or disorder (including obesity and diabetes), or macular degeneration (e.g., wet macular degeneration, e.g., wet age-related macular degeneration, e.g., the IL-18 mimetics can be used as an anti-angiogenic— as an illustrative example in some cases an IL-18 mimetics can attenuate choroidal neovascularization). Thus, methods are provided that include administering an IL-18 mimetic to treat or prevent a disease or disorder, such as, but not limited to, cancer, or infectious disease, a metabolic disease or disorder, or macular degeneration (e.g., wet macular degeneration such as wet age-related macular degeneration).

[0099] A method may include administering to a subject in need thereof a composition comprising at least one IL-18 mimetic. In some embodiments, a method comprises administering to a subject in need thereof a composition comprising at least one IL-18 mimetics, and a composition comprising an additional agent. In some such embodiments, the additional agent comprises an immunotherapeutic agent, e.g., an altered T-cell, a chimeric antigen receptor T-cell (CAR-T), an armored CAR-T cell, a virus, an antigen, a vaccine, an antibody, an immune checkpoint inhibitor, a small molecule, a chemotherapeutic agent, a stem cell, and the like. In some embodiments, a composition comprising at least one IL-18 mimetic is used in a method to increase immune system activity before, during, or after infection by a bacterium, virus, or other pathogen. In some embodiments, a composition comprising at least one IL-18 mimetic is used in a method to increase the number and / or activity of immune cells in vitro, in vivo or ex vivo, such as the number and / or activity of T cells, NK cells, and / or myeloid cells.

[0100] One of skill in the art will appreciate that an IL-18 mimetic of the present disclosure can be administered acutely (e.g., over a short period of time, such as a day, a week or a month) or chronically (e.g., over a long period of time, such as several months or a year or more). One of skill in the art will appreciate that the IL-18 mimetic can be administered singly or in any combination with other agents which may be administered concurrently, and / or before, and / or after each other.

[0101] 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 myeloidAtty Dkt. No.: MOBI-016WO 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, central nervous system lymphoma, cerebellar astrocytoma, cerebral malignant glioma, cerebral astrocytoma / malignant glioma, cervical cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, Ewing family of tumors, extracranial cancer, extrahepatic bile duct cancer, extrahepatic cancer, eye cancer, fungoides, gallbladder cancer, gastric (stomach) cancer, gastrointestinal cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, 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, kidney (renal cell) cancer, Langerhans cell cancer, laryngeal cancer, leukemia, lip and oral cavity cancer, liver cancer, lung cancer, lymphoma, macroglobulinemia, osteosarcoma, medulloblastoma, Merkel cell carcinoma, mesothelioma, mouth cancer, multiple endocrine neoplasia syndrome, myeloid leukemia, myeloma, myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumor, plasma cell neoplasm, pleuropulmonary blastoma, primary central nervous system cancer, prostate cancer, rectal cancer, renal cell (kidney) cancer, sarcoma, skin carcinoma, small cell lung cancer, small intestine cancer, squamous cell carcinoma, T-cell lymphoma, testicular cancer, throat cancer, thyroid cancer, trophoblastic tumor, urethral cancer, uterine cancer, vaginal cancer, and Wilms Tumor.

[0102] 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.

[0103] In some embodiments, the methods of the present disclosure are useful for treating or preventing a tumor or cancer that is resistant to immune checkpoint inhibitors (ICIs). Exemplary immune checkpoint inhibitors include, but is not limited to, anti-PDl (e.g., nivolumab), anti-CTLA4 (e.g., ipilimumab), anti-TIM3, anti-TIGIT, anti-LAG3, anti-B7H3, anti-B7H4, anti-VISTA, anti-ICOS, anti-GITR, anti-41BB, anti-OX40, and anti-CD40. Examples of targets of immune checkpoint inhibitors include but are not limited to: PD-L1, PD1, CTLA4, TIM3, TIGIT, LAG3, B7H3, B7H4, VISTA, ICOS, GITR, 41BB, 0X40, and CD40. Thus, examples of immune checkpoint inhibitors include agents that inhibit proteins such as: PD-L1, PD1, CTLA4, TIM3, TIGIT, LAG3, B7H3, B7H4, VISTA, ICOS, GITR, 41BB, 0X40, or CD40. In someAtty Dkt. No.: MOBI-016WO cases, a subject IL-18 mimetic is co-administered with an immune checkpoint inhibitor (e.g., an agent that inhibits PD-L1, PD1, CTLA4, TIM3, TIGIT, LAG3, B7H3, B7H4, VISTA, ICOS, GITR, 41BB, 0X40, or CD40, or any combination thereof).

[0104] Metabolic diseases and disorders include various metabolic and endocrine-related diseases and disorders. The following are non-limiting examples of metabolic and endocrine-related diseases and disorders that can be treated or prevented by the methods and compositions of the disclosure: obesity, diabetes, prediabetes, type II diabetes, mature onset diabetes of the young (MODY), hyperglycemia, dyslipidemia, hypertriglyceridemia, and hypercholesterolemia.

[0105] Non-limiting examples of other diseases and disorders that can be treated or prevented using the compositions and methods of the disclosure include viral infections, bacterial infections, parasitic infections, and low immune activity. In some embodiments, the viral infection is at least one of a pox virus, a smallpox virus, HPV infection, and warts caused by a virus. In some embodiments, the infection is a systemic infection. In some embodiments, the viral infection is a vaccinia virus infection. In some embodiments, the viral infection is a systemic vaccinia virus infection. In some embodiments, the bacterial infection is sepsis. In some embodiments, the low immune activity is neutropenia, for example, as may occur with chemotherapy.

[0106] Non-limiting examples of other diseases and disorders that can be treated or prevented using the compositions and methods of the disclosure include macular degeneration. For example, in some cases the disease or disorder is wet macular degeneration, and in some cases the disease or disorder is wet age-related macular degeneration. In some such cases, the IL-18 mimetics can be used as an anti-angiogenic, e.g., to attenuate choroidal neovascularization.

[0107] Thus, the present disclosure relates to the prevention and treatment of a disease or disorder by administration of a therapeutically effective amount of an IL-18 mimetic, a nucleic acid encoding the IL-18 , mimetic (e.g., DNA, cDNA, mRNA, etc.), a cell expressing the mimetic to a cell, tissue, or organ of a subject in need thereof, for the treatment or prevention of a disease or disorder, or its associated signs, symptoms or pathologies.

[0108] One skilled in the art, based upon the disclosure provided herein, would understand that the invention is useful in subjects who, in whole (e.g., systemically) or in part (e.g., locally, cell, tissue, organ), are being or will 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 hereinAtty Dkt. No.: MQBI-016WQ encompass any disease or disorder wherein an increase in IL-18 signaling will promote a positive biologic, physiological, clinical or therapeutic outcome.

[0109] Any suitable route of administration can be used for the treatment or prevention of a disease or disorder and can include parenteral administration, such as, cutaneous, subcutaneous, intraperitoneal, intravenous, intramuscular, intracisternal injection, kidney dialytic infusion techniques, etc.EXAMPLES OF NON-LIMITING ASPECTS OF THE DISCLOSURE

[0110] 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 the individually 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:1. An interleukin-18 (IL-18) mimetic comprising an amino acid sequence that is at least 80% identical to: >MBIO-7501 RYFGKLETKKVVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISAILLKDSQPRGMTVILVPEKEAPKL EKELEGKTVTREELEEIFKKLGIKAKLVRVSSVPGHDNKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSI MFTVEEVE (SEQ ID NO:1), >MBIO-8009RYFGKLETKKVVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISAIKLSDSLARGLTVILVPEKEAPKLE KELEGKTVTREELEEIFKKLGIKAKLVRVSDVPGHSRKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIM FTVEEVE (SEQ ID NO:21), >MBIQ-8010RYFGKLETKKVVIRNLNDQVLFLDQENRWLFEDMTDSDIRDNAGRVIISAIKLSDSLARGLTVILVPEKEAPKLE KELEGKTVTREELEEIFKKLGIKAKLVRVSDVPGHSRKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIM FYVEEVE (SE ID NO:22), >MBIQ-8011Atty Dkt. No.: MQBI-016WQRYFGKLETKKSVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISIIKLSDSLARGLTVIIVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRQSDVPGHSRKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIM FTVEEVE (SEQ ID NO:23),>MBIQ-8012RYFGKLETKKSVIRNLNDQVLFLDQENRWLFEDMTDSDIRDNAGRVIISIIKLSDSLARGLTVIIVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRQSDVPGHSRKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIM FYVEEVE (SEQ ID NO:24),>MBIQ-7505YFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHMYKDSQPRGMAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQM FYLEFED (SEQ ID N0:2),>MBIO_8168YFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHMYKDSQPRGMAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQM FYLEFED (SEQ ID N0:20),>MBIQ-8013YFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHKYSDSLARGLAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRDVPGHSRKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQMFYLEFED (SEQ ID NO:26),>MBIQ-8014YFGFLESKKSIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHKYSDSLARGLAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRDVPGHSRKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQMFYLENED (SEQ ID NO:27),>MBIO-7499MYFGKLEDRLVVIRNLNDQVLFLDDDGNLLFEDMTDSDIRDNAGRTVFRMTLLKDSQPRGMLVIFLPLDVVIKDTELRLVSHTDIWHTIEDSSGQLYQGIVAYVESVPGHDNKMRFRDAAKPDYYYGVKKERDLFKLALFKKPKLGDRSIMFTIEDVE (SEQ ID N0:3),>MBIQ-7500EKRYFGKLETSKVVIRNLNDQVLFRADDGSWLFEDMTDSDIRDNAGRTEILVTKLKDSQPRGMVVLLSKEEFDEDFVLEYDKETGKRTLISNGKELDFKDYLLVRVSSVPGHDNKMIFTSILTDGAYLAVKKERDLFKLVEVKNPKLGDRSIMFTVEPV (SEQ ID N0:4),>MBIO-7504Atty Dkt. No.: MOBI-016WOYFGFLESRKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWTIHMYKDSQPRGMAVTLSVNVEGWK TLSAKNRKVSFVEQPPPKNIKSDTSDIIFIQRSVPGHDNKMQFESSSYPGYFLAAVKERDLFKLVLKKREKLGDR NIMFYIEFED (SEQ ID N0:5), >MBIO-7506 YFGKLESKDAIIRNLNDQVLFIDQENRLLFEDMTDSDIRDNLPRTRFKIHMYKDSQPRGMAVTISVEVDKPYLL SFENRKVSFKEEEVPENIKSDTSDNIFFQRSVPGHDNKMQFESSSYPGYFLAVEKERDLFKLILKKKEKLGDRSI MFYLEFED (SEQ ID N0:6), >MBIO-7507YFGKLESKKAIIRNLNDQVLFIDQENRLLFEDMTDSDIRDNAPRTYFTIHMYKDSQPRGMAVTISVNVDKPYLL SAENGEISFKEMEPPENIKSDKSDIIFFQRSVPGHDNKMQFESSSYPGYFLAVEKERDLFKULKKREKLGDRSQ MFFLEFED (SEQ ID N0:7),>MBIO-7508YFGKLESRKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTTFNIHMYKDSQPRGMAVTISVNVDKPYL LSAENGKVSFKEEEVPENIKSDKSDNIFFQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKLILKKREKLGDRS QM FFLEFED (SEQ ID N0:8), >MBIO-7509YFGKLESRKVIIRNLNDQVLFIDQEGRALFEDMTDSDIRDNAPRTTFTIHMYKDSQPRGMAVTISVNVDKPYTL SAENKEVSFKEEEPPENIKEDESDIIFFQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKULKKKEKLGDRSQ MFYLEFED (SEQ ID N0:9), or>MBIO-7510YFGKLESKKAIIRNLNDQVLFIDQENRLLFEDMTDSDIRDNLPRTTFTIHMYKDSQPRGMAVAISVNVDKPYLL SFKNKKVEFKEEEVPENIKSDKSDIIFFQRSVPGHDNKMQFESSSYPGYFLAVEKERDLFKULKKKEKLGDRSIM FYLEFED (SEQ ID N0:10), wherein the IL-18 mimetic binds to human IL-18 receptor ( IL-18R). The IL-18 mimetic of aspect 1, wherein the amino acid sequence is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 1. The IL-18 mimetic of aspect 1, wherein the amino acid sequence is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 2.Atty Dkt. No.: MQBI-016WQ The IL-18 mimetic of aspect 1, wherein the amino acid sequence is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to one of SEQ ID NOs: 3-10 and 20-27. The IL-18 mimetic of any one of aspects 1-4, wherein the IL-18 mimetic has a higher solubility than a recombinant human IL-18 having the amino acid sequence set forth in SEQ ID NO:11. The IL-18 mimetic of any one of aspects 1-5, wherein the IL-18 mimetic has a higher melting temperature than a recombinant human IL-18 having the amino acid sequence set forth in SEQ ID NO:11. The IL-18 mimetic of any one of aspects 1-6, wherein the IL-18 mimetic does not significantly bind to IL-18 binding protein (IL-18BP). The IL-18 mimetic of aspect 7, wherein the IL-18 mimetic does not significantly bind to IL-18BP and comprises an amino acid sequence at least 80%, at least 85%, at least 90%, or at least 95% identical to the amino acid sequence of SEQ ID NO:1 and comprises 1, 2, 3, 4, 5, 6, 7, or 8 substitutions relative to the amino acid sequence of SEQ ID NO:1, wherein the substitutions are at positions 52, 54, 57, 58, 61, 106, 111, and / or 112, numbered relative to SEQ ID NO:1, optionally wherein the following amino acids K, S, L, A, L, D, S, and / or R are substituted in at positions 52, 54, 57, 58, 61, 106, 111, and / or 112, respectively. The IL-18 mimetic of aspect 8, wherein the substitutions are L52K, K54S, Q57L, P58A, M61L, S106D, D111S, and / or N112R, numbered relative to SEQ ID NO:1. The IL-18 mimetic of aspect 8 or 9, wherein the IL-18 mimetic that does not significantly bind to IL-18BP comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical to the amino acid sequence of:>MBIQ-8009 (SEQ ID NO:21), >MBIQ-8010 (SEQ ID NO:22), >MBIQ-8011 (SEQ ID NO:23), or >MBIQ-8012 (SEQ ID NO:24).Atty Dkt. No.: MQBI-016WQ The IL-18 mimetic of aspect 7 , wherein the IL-18 mimetic does not significantly bind to IL-18BP and comprises an amino acid sequence at least 80%, at least 85%, at least 90%, or at least 95% identical to the amino acid sequence of SEQ ID NO:2 and comprises 1, 2, 3, 4, 5, 6, 7, or 8 substitutions relative to the amino acid sequence of SEQ ID NO:2, wherein the substitutions are at positions 52, 54, 57, 58, 61, 106, 111, and / or 112, numbered relative to SEQ ID NO:1, optionally wherein the following amino acids K, S, L, A, L, D, S, and / or R are substituted in at positions 52, 54, 57, 58, 61, 106, 111, and / or 112, respectively. The IL-18 mimetic of aspect 11, wherein the substitutions are M52K, Y54S, Q57L, P58A, M61L, S106D, D111S, and / or N112R, numbered relative to SEQ ID NO:1. The IL-18 mimetic of aspect 11 or 12, wherein the IL-18 mimetic that does not significantly bind to IL-18BP comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical to the amino acid sequence of:>MBIQ-8013 (SEQ ID NO:26) or >MBIQ-8014 (SEQ ID NO:27). The IL-18 mimetic of any one of aspects 1-13 conjugated to a moiety. The IL-18 mimetic of aspect 14, wherein the moiety comprises one or more of an amino acid sequence, nucleic acid sequence, and a small molecule. The IL-18 mimetic of aspect 14 or 15, wherein the moiety comprises an immunoglobulin G (IgG) Fc sequence. The IL-18 mimetic of aspect 14 or 15, wherein the moiety comprises an antibody. The IL-18 mimetic of aspect 17, wherein the moiety comprises an scFv. The IL-18 mimetic of aspect 14 or 15, wherein the moiety comprises PEG. The IL-18 mimetic of aspect 14 or 15, wherein the moiety comprises a sugar molecule, e.g., a glycan.Atty Dkt. No.: MOBI-016WO The IL-18 mimetic of any one of aspects 1-20, wherein the IL-18 mimetic is conjugated to an immune checkpoint inhibitor, an immune agonist, or a cancer cell opsonizing agent. A nucleic acid comprising a nucleotide sequence encoding the IL-18 mimetic of any one of aspects 1-21. The nucleic acid of aspect 22, wherein the nucleic acid is a plasmid or a viral vector. A cell comprising the nucleic acid of aspect 22 or 23. The cell of aspect 24, wherein the cell is a bacterial cell or a yeast cell. The cell of aspect 24, wherein the cell is an engineered T or NK cell, a chimeric antigen receptor T cell (CAR-T cell), an engineered TCR-T cell, a chimeric antigen receptor NK cell (CAR-NK cell), or a Tumor-infiltrating Lymphocyte (TIL), that expresses the IL-18 mimetic. A pharmaceutical composition comprising the IL- 18 mimetic of any one of aspects 1-21, the nucleic acid of aspect 22 or 23, the cell of any one of aspects 24-26; and a pharmaceutically acceptable excipient. The pharmaceutical composition of aspect 27, further comprising:(i) an immune checkpoint inhibitor that inhibits PD-L1, PD1, CTLA4, TIM3, TIGIT, LAG3, B7H3, B7H4, VISTA, BTLA, CD47, SIRP alpha, CD48, CD155, CD160, TREM2, IDO1, Adenosine 2A receptor, Aryl hydrocarbon receptor, KIR, LILRB2, or any combination thereof;(ii) an immune agonist selected from: an agent that agonizes a tumor necrosis factor receptor superfamily (TNFRSF) protein (e.g., GITR, 4 IBB, 0X40, CD27, CD40, HVEM); an agent that agonizes an immunoglobulin superfamily (IgSF) protein (e.g., CD28, ICOS, CD226, NKG2D); an agent that agonizes a TLR (e.g., TLR2, TLR4, TLR5, TLR7, TLR9); an agent that agonizes a nucleic acid sensor (e.g., STING, cGAS, RIG-1 (DDX58)); an inflammasome activator; a T cell engager; a cytokine or cytokine variant (e.g., IL-2, IL-7, IL-10, IL-12, IL-15, IL-21, IL-23, IL-27, IL-33, TNF,TL1 A,IFNA,IFNB,IFNG); or any combination thereof;(iii) a cancer cell opsonizing agent that targets one or more antigens selected from: CD19, CD20, CD22, CD24, CD25, CD30, CD33, CD37, CD38, CD44, CD45, CD47, CD51, CD52, CD56, CD62L, CD70, CD74, CD79, CD80, CD96, CD97, CD99, CD123, CD134, CD138, CD152 (CTLA-4), CD200, CD213A2, CD221, CD248, CD276 (B7-H3), B7-H4, CD279 (PD-1), CD274 (PD-L1), CD319, EGFR, EPC AM, 17-1 A, HER1, HER2, HER3, CD 117, C-Met, HGFR,Atty Dkt. No.: MOBI-016WOPDGFRA, AXL, TWEAKR, PTHR2, HAVCR2 (TIM3), GD2 ganglioside, MUC1, mucin CanAg, mesothelin, endoglin, Lewis-Y antigen, CEA, CEACAM1, CEACAM5, CA-125, PSMA, BAFF, FGFR2, TAG-72, gelatinase B, glypican 3, nectin-4, BCMA, CSF1R, SLAMF7, integrin av 3, TYRP1, GPNMB, CLDN18.2, FOLR1, CCR4, CXCR4, MICA, C242 antigen, DLL3, DLL4, EGFL7, vimentin, fibronectin extra domain-B, TROP-2, LRRC15, FAP, SLITRK6, NOTCH2, NOTCH3, Tenascin-3, STEAP1, and NRPI; or(iv) an engineered T or NK cell, a chimeric antigen receptor T cell (CAR-T cell), an engineered TCR-T cell, a chimeric antigen receptor NK cell (CAR-NK cell), or a Tumor-infiltrating Lymphocyte (TIL). A kit comprising the IL-18 mimetic of any one of aspects 1-21, the nucleic acid of aspect 22 or 23, the cell of any one of aspects 24-26, and instructions for use. An in vitro or in vivo method of increasing IL-18 activity, the method comprising contacting I L18 with the IL-18 mimetic of any one of aspects 1-21. A method of increasing IL-18 activity in a subject in need thereof, the method comprising administering to the subject the IL- 18 mimetic of any one of aspects 1-21, the nucleic acid of aspect 22 or 23, the cell of any one of aspects 24-26; or the pharmaceutical composition of aspect 27 or 28. The method of aspect 31, wherein the subject has cancer, a metabolic disease, or an infectious disease. The method of aspect 32, wherein the cancer is a cancer that is resistant to immune checkpoint inhibitors.Atty Dkt. No.: MOBI-016WOEXAMPLES

[0111] The following examples are offered to illustrate, but not to limit any embodiments provided by the present disclosure.Example 1: IL-18 Mimetics Design

[0112] Design approach 1, native residues unmodified in the designs: 1 2 3 5 6 13 14 15 16 17 18 20 21 22 23 24 27 29 30 31 32 33 3435 36 37 39 4041 4449 51 52 53 54 55 56 57 58 59 606162 66 83 85 89 91 92 93 100 101 103 104 105 106 107 108 109 110 111 112 113 114 115 116 118 119 122 124 125 126 129 130 131 132 133 134 135 136 138 139 144 145 146 147 150 157.

[0113] Design approach 2 (allows generation of additional secondary structure elements): native residues unmodified in the designs: 1, 2, 3, 4, 5, 6, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 29, 30, 31, 32, 33, 34, 35, 36, 37, 39, 40, 41, 42, 53, 54, 55, 56, 57, 58, 59, 60, 105, 106, 107, 108, 109, 110, 111, 112, 113, 129, 130, 131, 132, 133, 134, 144, 145, 146, 147, 148, 149, 150, 151, and 152.

[0114] The main residues contacting the IL-18 receptors were maintained (see Design approach 1 and 2). The remainder of the residues were stabilized by optimizing secondary structure interactions.Additional secondary structure elements non-existent on the native IL-18 structure were added to stabilize the protein fold.

[0115] Native-IL-18 sequence is provided in SEQ ID NO:11. In-house expression of native IL-18 was unsuccessful as majority of the expressed protein was insoluble. Native IL-18 (recombinant human IL-18) was obtained from a commercial source (Bio-Techne Corporation).Table 1: Screening for IL-18 MimeticsAtty Dkt. No.: MOBI-016WOExample 2: IL-18 Mimetics Bind to IL-18BP

[0116] Table 2 shows binding of IL-18 mimetic, MBIO-7505 and MBIO-7501, to IL-18BP and the binding of rhlL-18 (Bio-Techne) to IL-18BP.

[0117] Table 2Atty Dkt. No.: MQBI-016WQExample 3: IL-18 Mimetics Activates IL-18 Signaling

[0118] FIG. 1 shows that IL-18 mimetics have signaling activity similar to or more potent than rhlL-18 as measured in HEK-Blue IL-18 reporter cells (Invivogen).Example 4: Decoy resistant IL-18 mimetics (IL-18 mimetics that do not bind IL-18BP protein)

[0119] The following amino acids, annotated by underlining, were substituted to generate IL-18 mimetics which have significantly reduced binding to IL-18BP. These mimetics are referred to as decoyresistant IL-18 mimetics. These decoy resistant IL-18 mimetics are expected to be more active in vivo, such as, in body fluids where IL-18BP is present and can bind and sequester IL-18, preventing it from binding and activating IL-18R.

[0120] >7505

[0121] YFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHMYKDSCJPRGMAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRS QM FYLEFED (SEQ ID NO:2)

[0122] >7501

[0123] RYFGKLETKKVVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISAILLKDSQPRGMTVILVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRVSSVPGHDNKM IFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSI M FTVEEVE (SEQ ID NO:1)

[0124] 7505-based decoy resistant IL-18 mimetics (the substituted-in amino acids are annotated by underlining):

[0125] >8013

[0126] YFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHKYSDSLARGLAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRDVPGHSRKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQ M FYLEFED (SEQ ID NO:26)

[0127] >8014

[0128] YFGFLESKKSIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHKYSDSLARGLAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRDVPGHSRKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQ M FYLENED (SEQ ID NO:27)

[0129] 7501-based decoy resistant IL-18 mimetics (the substituted-in amino acids are annotated by underlining):

[0130] >8009Atty Dkt. No.: MQBI-016WQ

[0131] RYFGKLETKKVVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISAIKLSDSLARGLTVILVPEK EAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRVSDVPGHSRKM IFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIM F TVEEVE (SEQ ID NO:21)

[0132] >8010

[0133] RYFGKLETKKVVIRNLNDQVLFLDQENRWLFEDMTDSDIRDNAGRVIISAIKLSDSLARGLTVILVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRVSDVPGHSRKM IFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIM F YVEEVE (SEQ ID NO:22)

[0134] >8011

[0135] RYFGKLETKKSVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISIIKLSDSLARGLTVIIVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRQSDVPGHSRKM IFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIMFT VEEVE (SEQ ID NO:23)

[0136] >8012

[0137] RYFGKLETKKSVIRNLNDQVLFLDQENRWLFEDMTDSDIRDNAGRVIISIIKLSDSLARGLTVIIVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRQS VPGHSRKM IFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIMFY VEEVE (SEQ ID NO:24)

[0138] Table 3 summarizes the binding kinetics of the listed IL-18 mimetics to IL-18 receptor complexes ( I L-18 Roc|3) and IL-18 binding protein. Binding measurements were performed by loading streptavidin (SA) biosensor probes with 50 nM biotinylated IL-18RP-Fc-Avi. Association was measured using a mixture of the IL-18 mimics at varying concentrations in the presence of 300 nM IL-18Ra-Fc-His, as well as in parallel assays against IL-18 binding protein (IL-18BP). Kinetic parameters, including association and dissociation rate constants and calculated equilibrium dissociation constants, were derived from global fitting of the sensorgrams.

[0139] Table 3Atty Dkt. No.: MOBI-016WO

[0140] FIGS. 5A-5C. Representative binding sensorgrams of wild-type IL-18 and IL-18 mimetics to IL-18 binding protein. Shown are representative biolayer interferometry ( BLI) sensorgrams illustrating binding of wild-type IL-18 (FIG. 5A) and the IL-18 mimetics MBIO-7505 (FIG. 5B), MBIO-7501 (FIG. 5C) to IL-18 binding protein (IL-18BP). Binding responses were recorded as changes in interference signal over time during the association and dissociation phases. These representative sensorgrams demonstrate differential binding behavior of the IL-18 mimetics relative to wild-type IL-18 and correspond to the kinetic and binding data described herein.

[0141] FIGS. 6A-6B. Representative binding sensorgrams of IL-18 mimetic MBIO-8013 to IL-18 receptor and IL-18 binding protein. Shown are representative biolayer interferometry (BLI) sensorgrams illustrating binding of the IL-18 mimetic MBIO-8013 to IL-18 binding protein (IL-18BP) (FIG. 6A) and to IL- 18 receptor (FIG. 6B). Binding interactions with IL-18R (in the presence of IL-18Ra) and with IL-18BP were monitored as changes in interference signal over time during the association and dissociation phases. These representative sensorgrams illustrate the binding profile of MBIO-8013 across IL-18 receptor and regulatory binding partners and correspond to the kinetic analyses described herein.

[0142] FIGS. 7A-7B. Biological activity of IL-18 mimetics in T-cell activation assays. The biological activity of IL-18 mimetics was evaluated by measuring interferon gamma (I FN-y) secretion from activated human T cells. Peripheral blood mononuclear cells (PBMCs) were expanded into T-cell blasts using anti-CD3 / anti-CD28 antibodies and interleukin-2 (IL-2), then treated with titrating concentrations of the IL-18 mimetics or wild-type IL-18 control (hrlL-18 (BioTechne). Culture supernatants were collected 12 hours after treatment and analyzed for IFN- y secretion. The IL-18 mimetics induced IFN- y secretion at levels comparable to or greater than wild-type IL-18, demonstrating preserved or enhanced biological activity.

[0143] FIG. 8. Thermal stability of IL-18 mimetics assessed by biolayer interferometry. The thermal stability of IL-18 mimetics MBIO-7501, MBIO-7505, and MBIO-8013 was evaluated relative to a recombinant human IL-18 control ("BT IL-18") following exposure to elevated temperatures. Proteins were incubated at either 65 °C or 80 °C for 30 minutes, followed by a 30-minute recovery period at roomAtty Dkt. No.: MOBI-016WO temperature. Binding activity after thermal stress was assessed by biolayer interferometry (BLI), where association response (association height) was used as a quantitative measure of functional binding. The IL-18 mimetics retained a greater proportion of binding activity following heat treatment compared to recombinant IL-18, demonstrating increased resistance to thermal denaturation.

[0144] FIGS. 9A-9B. Long-term thermal stability and retention of activity of IL-18 mimetics in cell-based assays. The long-term thermal stability and functional activity of the IL-18 mimetics were evaluated by measuring interferon-y (IFN-y) secretion following extended incubation under physiological conditions. IL-18 mimetics MBIO-8168 and MBIO-7505, along with a recombinant human IL-18 control (rhlL-18), were incubated in cell culture media at 37 °C for up to one month prior to functional testing. Activated human T-cell blasts, generated from peripheral blood mononuclear cells (PBMCs) expanded using anti-CD3 / anti-CD28 antibodies and interleukin-2 (IL-2), were treated with titrating concentrations of the pre-incubated proteins. Culture supernatants were collected 12 hours after treatment and analyzed for IFN-y secretion by ELISA. The rhl L-18IL-18 control exhibited a marked loss of biological activity over time (FIG. 9A), whereas MBIO-8168 (FIG. 9A) and MBIO-7505 (FIG. 9B) retained comparable IFN-y-inducing activity throughout the one-month incubation period, demonstrating enhanced thermostability and sustained functional potency of the IL-18 mimetics.

[0145] While the subject proteins have been particularly shown and described with references to certain embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

Claims

Atty Dkt. No.: MQBI-016WQCLAIMSWhat is claimed is:

1. An interleukin-18 (IL-18) mimetic comprising an amino acid sequence that is at least 80% identical to:>MBIO-7501RYFGKLETKKVVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISAILLKDSQPRGMTVILVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRVSSVPGHDNKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIMFTVEEVE (SEQ ID NO:1),>MBIQ-8009RYFGKLETKKVVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISAIKLSDSLARGLTVILVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRVSDVPGHSRKMIFTLLGKEEKYLA VKERDLFVLKEVKKVKLGDRSIMFTVEEVE (SEQ ID NO:21),>MBIO-8010RYFGKLETKKVVIRNLNDQVLFLDQENRWLFEDMTDSDIRDNAGRVIISAIKLSDSLARGLTVILVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRVSDVPGHSRKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIMFYVEEVE (SEQ ID NO:22),>MBIO-8011RYFGKLETKKSVIRNLNDQVLFLADDGTWLFEDMTDSDIRDNAGRVIISIIKLSDSLARGLTVIIVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRQSDVPGHSRKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIM FTVEEVE (SEQ ID NO:23),>MBIO-8012RYFGKLETKKSVIRNLNDQVLFLDQENRWLFEDMTDSDIRDNAGRVIISIIKLSDSLARGLTVIIVPEKEAPKLEKELEGKTVTREELEEIFKKLGIKAKLVRQSDVPGHSRKMIFTLLGKEEKYLAVVKERDLFVLKEVKKVKLGDRSIIVI FYVEEVE (SEQ ID NO:24),>MBIO-7505YFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHMYKDSQPRGIVIAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQM FYLEFED (SEQ ID NO:2),>MBIO_8168Atty Dkt. No.: MQBI-016WQYFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHMYKDSQPRGMAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQM FYLEFED (SEQ ID N0:20),>MBIO-8013YFGFLESKKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHKYSDSLARGLAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRDVPGHSRKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQMFYLEFED (SEQ ID NO:26),>MBIO-8014YFGFLESKKSIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWYIHKYSDSLARGLAVTISVKVDGWKTLSAKNKKVEFVEQKPPKNIKSDKSDIIFIQRDVPGHSRKMQFESSSYPGYFLAAEKERDLFKLVLKKKEKLGDRSQMFYLENED (SEQ ID NO:27),>MBIO-7499MYFGKLEDRLVVIRNLNDQVLFLDDDGNLLFEDMTDSDIRDNAGRTVFRMTLLKDSQPRGMLVIFLPLDVVIKDTELRLVSHTDIWHTIEDSSGQLYQGIVAYVESVPGHDNKMRFRDAAKPDYYYGVKKERDLFKLALFKKPKLGDRSIMFTIEDVE (SEQ ID N0:3),>MBIQ-7500EKRYFGKLETSKVVIRNLNDQVLFRADDGSWLFEDMTDSDIRDNAGRTEILVTKLKDSQPRGMVVLLSKEEFDEDFVLEYDKETGKRTLISNGKELDFKDYLLVRVSSVPGHDNKMIFTSILTDGAYLAVKKERDLFKLVEVKNPKLGDRSIMFTVEPV (SEQ ID N0:4),>MBIO-7504YFGFLESRKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTYWTIHMYKDSQPRGMAVTLSVNVEGWKTLSAKNRKVSFVEQPPPKNIKSDTSDIIFIQRSVPGHDNKMQFESSSYPGYFLAAVKERDLFKLVLKKREKLGDRNIMFYIEFED (SEQ ID N0:5),>MBIO-7506YFGKLESKDAIIRNLNDQVLFIDQENRLLFEDMTDSDIRDNLPRTRFKIHMYKDSQPRGMAVTISVEVDKPYLLSFENRKVSFKEEEVPENIKSDTSDNIFFQRSVPGHDNKMQFESSSYPGYFLAVEKERDLFKLILKKKEKLGDRSIMFYLEFED (SEQ ID N0:6),>MBIO-7507YFGKLESKKAIIRNLNDQVLFIDQENRLLFEDMTDSDIRDNAPRTYFTIHMYKDSQPRGMAVTISVNVDKPYLLSAENGEISFKEMEPPENIKSDKSDIIFFQRSVPGHDNKMQFESSSYPGYFLAVEKERDLFKULKKREKLGDRSQMFFLEFED (SEQ ID N0:7),>MBIO-7508Atty Dkt. No.: MOBI-016WOYFGKLESRKAIIRNLNDQVLFIDQENRALFEDMTDSDIRDNAPRTTFNIHMYKDSQPRGMAVTISVNVDKPYL LSAENGKVSFKEEEVPENIKSDKSDNIFFQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKLILKKREKLGDRS QM FFLEFED (SEQ ID N0:8), >MBIO-7509 YFGKLESRKVIIRNLNDQVLFIDQEGRALFEDMTDSDIRDNAPRTTFTIHMYKDSQPRGMAVTISVNVDKPYTL SAENKEVSFKEEEPPENIKEDESDIIFFQRSVPGHDNKMQFESSSYPGYFLAAEKERDLFKULKKKEKLGDRSQ MFYLEFED (SEQ ID N0:9), or >MBIO-7510 YFGKLESKKAIIRNLNDQVLFIDQENRLLFEDMTDSDIRDNLPRTTFTIHMYKDSQPRGMAVAISVNVDKPYLL SFKNKKVEFKEEEVPENIKSDKSDIIFFQRSVPGHDNKMQFESSSYPGYFLAVEKERDLFKULKKKEKLGDRSIM FYLEFED (SEQ ID N0:10), wherein the IL-18 mimetic binds to human IL-18 receptor ( IL-18R).

2. The IL-18 mimetic of claim 1, wherein the amino acid sequence is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 1.

3. The IL-18 mimetic of claim 1, wherein the amino acid sequence is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 2.

4. The IL-18 mimetic of claim 1, wherein the amino acid sequence is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to one of SEQ ID NOs: 3-10 and 20-27.

5. The IL-18 mimetic of any one of claims 1-4, wherein the IL-18 mimetic has a higher solubility than a recombinant human IL-18 having the amino acid sequence set forth in SEQ ID NO:11.

6. The IL-18 mimetic of any one of claims 1-5, wherein the IL-18 mimetic has a higher melting temperature than a recombinant human IL-18 having the amino acid sequence set forth in SEQ ID NO:11.Atty Dkt. No.: MQBI-016WQ7. The IL-18 mimetic of any one of claims 1-6, wherein the IL-18 mimetic does not significantly bind to IL-18 binding protein (IL-18BP).

8. The IL-18 mimetic of claim 7, wherein the IL-18 mimetic does not significantly bind to IL-18 BP and comprises an amino acid sequence at least 80%, at least 85%, at least 90%, or at least 95% identical to the amino acid sequence of SEQ ID NO:1 and comprises 1, 2, 3, 4, 5, 6, 7, or 8 substitutions relative to the amino acid sequence of SEQ ID NO:1, wherein the substitutions are at positions 52, 54, 57, 58, 61, 106, 111, and / or 112, numbered relative to SEQ ID NO:1.

9. The IL-18 mimetic of claim 8, wherein the substitutions are L52K, K54S, Q57L, P58A, M61L, S106D, D111S, and / or N112R, numbered relative to SEQ ID NO:1.

10. The IL-18 mimetic of claim 8 or 9, wherein the IL-18 mimetic that does not significantly bind to IL-18BP comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical to the amino acid sequence of:>MBIQ-8009 (SEQ ID NO:21), >MBIQ-8010 (SEQ ID NO:22), >MBIQ-8011 (SEQ ID NO:23), or >MBIQ-8012 (SEQ ID NO:24).

11. The IL-18 mimetic of claim 7, wherein the IL-18 mimetic does not significantly bind to IL-18 BP and comprises an amino acid sequence at least 80%, at least 85%, at least 90%, or at least 95% identical to the amino acid sequence of SEQ ID NO:2 and comprises 1, 2, 3, 4, 5, 6, 7, or 8 substitutions relative to the amino acid sequence of SEQ ID NO:2, wherein the substitutions are at positions 52, 54, 57, 58, 61, 106, 111, and / or 112, numbered relative to SEQ ID NO:1.

12. The IL-18 mimetic of claim 11, wherein the substitutions are M52K, Y54S, Q57L, P58A, M61L, S106D, D111S, and / or N112R, numbered relative to SEQ ID NO:1.

13. The IL-18 mimetic of claim 11 or 12, wherein the IL-18 mimetic that does not significantly bind to IL-18BP comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical to the amino acid sequence of:>MBIQ-8013 (SEQ ID NO:26) or >MBIQ-8014 (SEQ ID NO:27).Atty Dkt. No.: MOBI-016WO14. The IL-18 mimetic of any one of claims 1-13 conjugated to a moiety.

15. The IL-18 mimetic of claim 14, wherein the moiety comprises one or more of an amino acid sequence, nucleic acid sequence, and a small molecule.

16. The IL-18 mimetic of claim 14 or 15, wherein the moiety comprises an immunoglobulin G (IgG) Fc sequence.

17. The IL-18 mimetic of claim 14 or 15, wherein the moiety comprises an antibody.

18. The IL-18 mimetic of claim 17, wherein the moiety comprises an scFv.

19. The IL-18 mimetic of claim 14 or 15, wherein the moiety comprises PEG.

20. The IL-18 mimetic of claim 14 or 15, wherein the moiety comprises a sugar molecule, e.g., a glycan.

21. The IL-18 mimetic of any one of claims 1-20, wherein the IL-18 mimetic is conjugated to an immune checkpoint inhibitor, an immune agonist, or a cancer cell opsonizing agent.

22. A nucleic acid comprising a nucleotide sequence encoding the IL-18 mimetic of any one of claims 1-21.

23. The nucleic acid of claim 22, wherein the nucleic acid is a plasmid or a viral vector.

24. A cell comprising the nucleic acid of claim 22 or 23.

25. The cell of claim 24, wherein the cell is a bacterial cell or a yeast cell.

26. The cell of claim 24, wherein the cell is an engineered T or NK cell, a chimeric antigen receptor T cell (CAR-T cell), an engineered TCR-T cell, a chimeric antigen receptor NK cell (CAR-NK cell), or a Tumor-infiltrating Lymphocyte (TIL), that expresses the IL-18 mimetic.

27. A pharmaceutical composition comprising the IL- 18 mimetic of any one of claims 1-21, the nucleic acid of claim 22 or 23, the cell of any one of claims 24-26; and a pharmaceutically acceptable excipient.

28. The pharmaceutical composition of claim 27, further comprising:Atty Dkt. No.: MOBI-016WO(i) an immune checkpoint inhibitor that inhibits PD-L1, PD1, CTLA4, TIM3, TIGIT, LAG3, B7H3, B7H4, VISTA, BTLA, CD47, SIRP alpha, CD48, CD155, CD160, TREM2, IDO1, Adenosine 2A receptor, Aryl hydrocarbon receptor, KIR, LILRB2, or any combination thereof;(ii) an immune agonist selected from: an agent that agonizes a tumor necrosis factor receptor superfamily (TNFRSF) protein (e.g., GITR, 4 IBB, 0X40, CD27, CD40, HVEM); an agent that agonizes an immunoglobulin superfamily (IgSF) protein (e.g., CD28, ICOS, CD226, NKG2D); an agent that agonizes a TLR (e.g., TLR2, TLR4, TLR5, TLR7, TLR9); an agent that agonizes a nucleic acid sensor (e.g., STING, cGAS, RIG-1 (DDX58)); an inflammasome activator; a T cell engager; a cytokine or cytokine variant (e.g., IL-2, IL-7, IL-10, IL-12, IL-15, IL-21, IL-23, IL-27, IL-33, TNF,TL1 A,IFNA,IFNB,IFNG); or any combination thereof;(iii) a cancer cell opsonizing agent that targets one or more antigens selected from: CD19, CD20, CD22, CD24, CD25, CD30, CD33, CD37, CD38, CD44, CD45, CD47, CD51, CD52, CD56, CD62L, CD70, CD74, CD79, CD80, CD96, CD97, CD99, CD123, CD134, CD138, CD152 (CTLA-4), CD200, CD213A2, CD221, CD248, CD276 (B7-H3), B7-H4, CD279 (PD-1), CD274 (PD-L1), CD319, EGFR, EPC AM, 17-1 A, HER1, HER2, HER3, CD 117, C-Met, HGFR, PDGFRA, AXL, TWEAKR, PTHR2, HAVCR2 (TIM3), GD2 ganglioside, MUC1, mucin CanAg, mesothelin, endoglin, Lewis-Y antigen, CEA, CEACAM1, CEACAM5, CA-125, PSMA, BAFF, FGFR2, TAG-72, gelatinase B, glypican 3, nectin-4, BCMA, CSF1R, SLAMF7, integrin av 3, TYRP1, GPNMB, CLDN18.2, F0LR1, CCR4, CXCR4, MICA, C242 antigen, DLL3, DLL4, EGFL7, vimentin, fibronectin extra domain-B, TROP-2, LRRC15, FAP, SLITRK6, NOTCH2, NOTCH3, Tenascin-3, STEAP1, and NRPI; or(iv) an engineered T or NK cell, a chimeric antigen receptor T cell (CAR-T cell), an engineered TCR-T cell, a chimeric antigen receptor NK cell (CAR-NK cell), or a Tumor-infiltrating Lymphocyte (TIL).

29. A kit comprising the IL-18 mimetic of any one of claims 1-21, the nucleic acid of claim 22 or 23, the cell of any one of claims 24-26, and instructions for use.

30. An in vitro or in vivo method of increasing IL-18 activity, the method comprising contacting I L18 with the IL-18 mimetic of any one of claims 1-21.

31. A method of increasing IL-18 activity in a subject in need thereof, the method comprising administering to the subject the IL- 18 mimetic of any one of claims 1-21, the nucleic acid ofAtty Dkt. No.: MOBI-016WO claim 22 or 23, the cell of any one of claims 24-26; or the pharmaceutical composition of claim 27 or 28.

32. The method of claim 31, wherein the subject has cancer, a metabolic disease, or an infectious disease.

33. The method of claim 32, wherein the cancer is a cancer that is resistant to immune checkpoint inhibitors.

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